FR2964834A1 - Antioxidant composition, useful as food supplement and to prevent and/or treat infertility, protect sperm and prevent spontaneous abortion/embryonic malformations, comprises prickly pear extract, vitamins B6, B9 and B12 and zinc - Google Patents
Antioxidant composition, useful as food supplement and to prevent and/or treat infertility, protect sperm and prevent spontaneous abortion/embryonic malformations, comprises prickly pear extract, vitamins B6, B9 and B12 and zinc Download PDFInfo
- Publication number
- FR2964834A1 FR2964834A1 FR1057556A FR1057556A FR2964834A1 FR 2964834 A1 FR2964834 A1 FR 2964834A1 FR 1057556 A FR1057556 A FR 1057556A FR 1057556 A FR1057556 A FR 1057556A FR 2964834 A1 FR2964834 A1 FR 2964834A1
- Authority
- FR
- France
- Prior art keywords
- extract
- antioxidant composition
- composition according
- prevent
- prickly pear
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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Abstract
Description
Domaine de l'invention La présente invention se rapporte au domaine du traitement de l'infertilité chez l'homme et chez la femme. En particulier, la présente invention concerne un complément alimentaire appliqué pour prévenir et/ou traiter l'infertilité chez l'Homme et, notamment l'infertilité conjugale liée au stress oxydatif. Etat de la technique Les causes d'une infertilité se recherchent à quatre niveaux : qualité du sperme, ovulations, voies génitales féminines et masculines, et incompatibilité entre le sperme et le milieu génital féminin. De nombreuses études ont permis de mettre en évidence le déclin de la qualité du sperme des hommes fertiles, depuis une vingtaine d'années, ce qui entraînerait une altération de la fécondité dans les années à venir. La question de l'infertilité conjugale doit faire prendre conscience de l'incidence sur la fertilité masculine et féminine des facteurs liés à l'environnement et des facteurs nutritionnels. Dans les pays industriels, 15 % des couples sont inféconds dont les causes identifiées sont diverses mais une grande proportion ne montre aucune anomalie au niveau de chacun des deux conjoints et cependant ils n'arrivent pas à concevoir un enfant : c'est l'infécondité inexpliquée. Field of the Invention The present invention relates to the field of the treatment of infertility in men and women. In particular, the present invention relates to a dietary supplement applied to prevent and / or treat infertility in humans and, especially marital infertility related to oxidative stress. STATE OF THE ART The causes of infertility are sought at four levels: sperm quality, ovulation, female and male genital tract, and incompatibility between semen and the female genital environment. Numerous studies have demonstrated the decline in sperm quality of fertile men over the last twenty years, which would lead to an alteration of fertility in the years to come. The issue of marital infertility must raise awareness of the impact on male and female fertility of environmental factors and nutritional factors. In industrialized countries, 15% of couples are infertile whose causes are identified but a large proportion show no anomaly at the level of each of the two spouses and yet they fail to conceive a child: it is infertility unexplained.
L'infécondité peut avoir pour origine l'action des agents toxiques, pesticides (dioxine avec effet oestrogénique par blocage des récepteurs aux androgènes), les métaux lourds, le tabac (par le biais des radicaux libres), l'alcool, les agents pharmacologiques, les rayons X et gamma, les hormones (absorbées par le bétail puis consommées ensuite par les humains), l'infection ou l'inflammation. Ces facteurs sont susceptibles d'entraîner l'apparition d'un stress oxydatif. Le stress oxydatif est, dans ce contexte, l'intermédiaire dans l'infécondité. Le stress oxydatif est le déséquilibre entre la production de radicaux libres et les défenses antioxydantes. En conditions normales, les radicaux libres sont produits en permanence dans l'organisme humain et détruits par les défenses antioxydantes. Ce processus est nocif pour l'organisme et représente une cause importante de dommages des structures cellulaires, dont les lipides et les membranes, les protéines et l'ADN. 1 A noter que le risque de la lipoperoxydation existe même chez des couples normalement fertiles, d'où l'importance de remédier au stress oxydatif. Le stress oxydatif est un facteur de risque d'une pathologie qui intervient dans environ la moitié des cas chez les hommes infertiles. Infertility can be caused by the action of toxic agents, pesticides (dioxin with estrogenic effect by blocking androgen receptors), heavy metals, tobacco (through free radicals), alcohol, pharmacological agents , X and gamma rays, hormones (absorbed by livestock and then consumed by humans), infection or inflammation. These factors are likely to lead to the onset of oxidative stress. Oxidative stress is, in this context, the intermediary in infertility. Oxidative stress is the imbalance between the production of free radicals and antioxidant defenses. Under normal conditions, free radicals are permanently produced in the human body and destroyed by antioxidant defenses. This process is harmful to the body and is a major cause of damage to cellular structures including lipids and membranes, proteins and DNA. It should be noted that the risk of lipoperoxidation exists even in normally fertile couples, hence the importance of remedying oxidative stress. Oxidative stress is a risk factor for a condition that occurs in about half of all cases in infertile men.
Les conséquences possibles du stress oxydatif sur la fécondité sont les suivantes : - Cassures au niveau des chromosomes altérant le message génétique (mesuré par l'index de fragmentation de l'ADN du spermatozoïde (DFI)), fragmentation de l'ADN pouvant être isolée ou associée à une décondensation de la chromatine (exprimée par l'indice de décondensation de l'ADN (DSI)); - Peroxydation des acides gras polyinsaturés des membranes cellulaires (identifiée par l'analyse des phospholipides membranaires) entraînant une atteinte de la motilité et de la capacité de fusionner du spermatozoïde, un déficit de la perméabilité et des troubles dans les échanges pouvant aller jusqu'à la mort cellulaire. Chez l'homme, le stress oxydatif va affecter: la concentration du sperme, la mobilité des spermatozoïdes (asthénospermie, moins de 40 % de spermatozoïdes mobiles), la morphologie des spermatozoïdes (tératospermie, moins de 30 % de spermatozoïdes normaux). The possible consequences of oxidative stress on fertility are as follows: - Breaks at the level of chromosomes altering the genetic message (measured by the sperm DNA fragmentation index (DFI)), fragmentation of the DNA can be isolated or associated with a decondensation of chromatin (expressed by the DNA decondensation index (DSI)); - Peroxidation of polyunsaturated fatty acids of cell membranes (identified by the analysis of membrane phospholipids) resulting in impaired motility and ability to fuse sperm, a deficit in permeability and disorders in trade up to cell death. In humans, oxidative stress will affect: sperm concentration, sperm motility (asthenospermia, less than 40% motile sperm), sperm morphology (teratospermia, less than 30% normal spermatozoa).
Les atteintes de la production des spermatozoïdes, de leur motilité, de leur trajectoire et de leur intégrité sont identifiées par le spermogramme et spermocytogramme. Chez la femme, le stress oxydatif peut entraîner sur l'ovocyte les mêmes inconvénients que pour le spermatozoïde (attaque de l'ADN et des acides gras polyinsaturés de la membrane). En outre, la femme est exposée à des risques spécifiques, selon le déficit en certains antioxydants. C'est ainsi que le déficit de sélénium (qui s'accompagne d'une baisse du glutathion peroxydase) pourrait entraîner une fausse couche par suite des dégâts oxydatifs directs sur les membranes et l'ADN du foetus. Impairment of sperm production, motility, trajectory and integrity are identified by spermogram and spermocytogram. In women, oxidative stress can cause the same problems for the oocyte as for the spermatozoon (attack of DNA and polyunsaturated fatty acids of the membrane). In addition, the woman is exposed to specific risks, depending on the deficit in certain antioxidants. For example, selenium deficiency (which is associated with a decrease in glutathione peroxidase) could lead to miscarriage as a result of direct oxidative damage to membranes and fetal DNA.
Des études sur les causes d'infertilité et leurs traitements ont déjà été réalisées et il est connu que des antioxydants tels que la carnitine, le zinc, la vitamine E, le coenzyme Q10 et la vitamine B12 agissent sur la motilité du sperme. Le zinc et la vitamine B12 agissent également sur la production du sperme. La vitamine C permet de protéger le patrimoine génétique du sperme. Le glutathion et le sélénium sont deux éléments essentiels pour la défense antioxydante du sperme, le glutathion permet également d'augmenter la motilité du sperme. Jusqu'à présent, ces différents traitements ont été utilisés isolément. Studies on the causes of infertility and their treatments have already been performed and it is known that antioxidants such as carnitine, zinc, vitamin E, coenzyme Q10 and vitamin B12 act on the motility of sperm. Zinc and vitamin B12 also act on sperm production. Vitamin C helps protect the gene pool of sperm. Glutathione and selenium are two essential elements for the antioxidant defense of sperm, glutathione also helps increase sperm motility. So far, these different treatments have been used in isolation.
L'invention a ainsi pour but de proposer une nouvelle composition ayant une action anti-oxydante afin de prévenir et/ou traiter l'infertilité chez l'Homme, tout au moins aussi efficace que celles existantes sur le marché à l'heure actuelle. The object of the invention is therefore to propose a new composition having an antioxidant action for preventing and / or treating infertility in humans, at least as effective as those currently on the market at the present time.
Description de l'invention A cet effet, la présente invention concerne une composition anti-oxydante comprenant comme principes actifs : un extrait de figues de Barbarie, des vitamines B6, B9, B12, et du zinc. Le présent demandeur a déterminé que l'emploi d'antioxydants conventionnels comme les vitamines A, C, E et du sélénium, présente un impact inconstant, à la fois sur : la survie et la motilité des spermatozoïdes, l'indice de fragmentation de l'ADN (DFI), et sur l'analyse des phospholipides membranaires. Ainsi, le simple fait d'utiliser des antioxydants ne suffit pas à rendre une composition efficace afin de prévenir et/ou de traiter l'infertilité chez l'Homme. La présente invention se rapporte ainsi à une sélection très particulière d'antioxydants apte à prévenir les problèmes d'infertilité, de fausses couches ou les malformations embryonnaires. Le présent demandeur a en effet découvert, de manière surprenante, que le mélange des composés actifs susmentionnés, à savoir un extrait de figue de Barbarie, des vitamines B6, B9 et B12 et du zinc, présente une activité synergique antioxydante afin de contribuer à la protection des constituants cellulaires contre les dommages oxydatifs, notamment réduire la fragmentation et la décondensation de l'ADN du spermatozoïde chez l'homme. Il a découvert que l'extrait de figues de Barbarie allié à l'action des vitamines B6, B12, B9 et du zinc réduit le risque de fragmentation de l'ADN et permet de réduire le taux d'homocystéine via la méthylation, et jouent un rôle dans le métabolisme et la formation du sperme chez l'homme. De plus, le zinc joue un rôle dans le développement testiculaire, notamment dans la transformation de la testostérone en DHT (dihydrotestostérone, forme active de la testostérone). Par conséquent, la composition selon la présente invention n'intervient plus, contrairement aux traitements existants, seulement sur la mobilité du sperme, mais agit également sur la protection des membranes du sperme, la protection du bagage génétique, la mobilité et la trajectoire du sperme et l'apport d'énergie. Une protection globale des spermatozoïdes est ainsi obtenue. DESCRIPTION OF THE INVENTION For this purpose, the present invention relates to an antioxidant composition comprising as active ingredients: an extract of prickly pears, vitamins B6, B9, B12, and zinc. The present applicant has determined that the use of conventional antioxidants such as vitamins A, C, E and selenium, has an inconsistent impact on both sperm survival and motility, fragmentation index of DNA (DFI), and on the analysis of membrane phospholipids. Thus, the mere fact of using antioxidants is not enough to make an effective composition to prevent and / or treat infertility in humans. The present invention thus relates to a very particular selection of antioxidants capable of preventing problems of infertility, miscarriage or embryonic malformations. The present applicant has in fact surprisingly discovered that the mixture of the abovementioned active compounds, namely an extract of prickly pears, vitamins B6, B9 and B12 and zinc, has a synergistic antioxidant activity in order to contribute to the protection of cellular constituents against oxidative damage, including reducing the fragmentation and decondensation of sperm DNA in humans. He discovered that prickly pear extract combined with the action of vitamins B6, B12, B9 and zinc reduced the risk of DNA fragmentation and reduced homocysteine levels via methylation, and a role in the metabolism and sperm formation in humans. In addition, zinc plays a role in testicular development, especially in the transformation of testosterone into DHT (dihydrotestosterone, active form of testosterone). Therefore, the composition according to the present invention does not intervene, unlike existing treatments, only on the mobility of sperm, but also acts on the protection of sperm membranes, protection of genetic background, mobility and trajectory of sperm and the supply of energy. Overall protection of spermatozoa is thus obtained.
De préférence, l'extrait de figue de Barbarie comprend, en poids par rapport au poids total de l'extrait de figue de Barbarie, de 0,10% à 25%, de préférence de 8% à 20% de bétalaïnes, de 0,10 % à 25%, de préférence de 8% à 15% de quercétine et 0,10% à 20%, de préférence de 6% à 15% de terpènes. Selon le présent demandeur, les bétalaïnes réduiraient en effet le risque de fragmentation de l'ADN et la peroxydation lipidique, tout comme la quercétine qui en outre protègerait la qualité du sperme chez l'homme, ainsi que les premiers stades de l'embryogénèse et donc aiderait à lutter contre la fausse couche et les malformations de l'embryon chez la femme. La quercétine aurait de plus une action protectrice sur la lignée germinale. Les terpènes quant à eux déclencheraient le mécanisme des HSP réparateurs de protéines. Le procédé afin d'obtenir un extrait de figues de Barbarie enrichi en bétalaïnes, quercétine et terpène selon l'invention comprend l'étape consistant à réaliser une extraction avec un solvant polaire à partir d'extrait de fruit d'Opuntia ficus indica afin d'en récupérer les bétalaïnes, la quercétine et les terpènes. Preferably, the prickly pear extract comprises, by weight relative to the total weight of the prickly pear extract, from 0.10% to 25%, preferably from 8% to 20% of betalaines, from , 10% to 25%, preferably 8% to 15% quercetin and 0.10% to 20%, preferably 6% to 15% terpenes. According to the present applicant, betalaines would indeed reduce the risk of DNA fragmentation and lipid peroxidation, as would quercetin, which would also protect the quality of sperm in humans, as well as the early stages of embryogenesis and therefore help to fight against miscarriage and malformations of the embryo in women. Quercetin would also have a protective effect on the germ line. Terpenes trigger the mechanism of protein repair HSPs. The process for obtaining an extract of prickly pearls enriched with betalaines, quercetin and terpene according to the invention comprises the step of carrying out an extraction with a polar solvent from fruit extract of Opuntia ficus indica in order to recover betalaines, quercetin and terpenes.
Par extrait, on entend une composition obtenue à partir d'une plante par une opération physique et/ou chimique, en particulier d'au moins une partie de la plante par au moins un solvant. Ce procédé comprend en particulier les étapes suivantes : de la figue de Barbarie est placée dans un solvant polaire, puis une étape d'extraction est réalisée et l'extrait obtenu est récupéré. Le solvant polaire est de préférence un solvant présentant un moment dipolaire différent de zéro et en particulier supérieur à 1,5. Le solvant polaire est avantageusement un solvant protique. Les alcools comme le méthanol, l'éthanol, le propanol, le butanol et le pentanol, l'eau comme l'eau osmosée, distillée, permutée conviennent comme solvant polaire pour réaliser le procédé. Le solvant peut comprendre en outre un additif tel qu'un agent antioxydant, un agent conservateur, une ou des enzymes, notamment destinées à dégrader les parois végétales, par exemple pour faciliter l'extraction, un agent anti-moussant, un acide ou encore une base, notamment pour modifier le point isoélectrique et favoriser la précipitation. L'étape d'extraction peut se faire par : décoction, macération, infusion, digestion, percolation, lixiviation, entraînement à l'eau chaude, comme la distillation ou hydrodistillation, solvant assisté par micro-ondes, et par colonne échangeuse d'ions. La décoction comprend les étapes suivantes : - la mise à ébullition d'un mélange comprenant des fruits de figuier de Barbarie dans un solvant polaire tel que défini ci-dessus ; et - le maintien de cette ébullition pendant une durée particulière, notamment de 1 minute à 2 heures, en particulier environ 20 minutes, puis éventuellement un retour à température ambiante. On entend par « macération », un procédé consistant à laisser tremper à froid les fruits de figuier de Barbarie dans le solvant polaire pendant une durée particulière, afin d'en extraire les composés solubles. En particulier, l'étape de macération a une durée allant de 1 heure à 15 jours, et de manière préférée de 1 jour. Tout particulièrement, elle se fait à température ambiante, soit de 10 à 25°C. L'infusion est un procédé consistant à verser le solvant polaire se trouvant de préférence à température d'ébullition sur les fruits de figuier de Barbarie, puis à laisser le mélange infuser pendant une période allant de une minute à 2 heures. La digestion est un procédé consistant à chauffer un mélange comprenant les fruits de figuier de Barbarie dans le solvant polaire à une température inférieure à la température d'ébullition, par exemple dans le cas de l'eau de 10 à 89°C, et en particulier de 25 à 70°C. By extract is meant a composition obtained from a plant by a physical and / or chemical operation, in particular at least a part of the plant with at least one solvent. This process comprises in particular the following steps: prickly pear is placed in a polar solvent, then an extraction step is performed and the resulting extract is recovered. The polar solvent is preferably a solvent having a dipole moment other than zero and in particular greater than 1.5. The polar solvent is advantageously a protic solvent. Alcohols such as methanol, ethanol, propanol, butanol and pentanol, water as osmosis water, distilled, permuted are suitable polar solvent for carrying out the process. The solvent may furthermore comprise an additive such as an antioxidant agent, a preserving agent, one or more enzymes, in particular intended to degrade the plant walls, for example to facilitate the extraction, an anti-foaming agent, an acid or else a base, especially to modify the isoelectric point and promote precipitation. The extraction step can be done by: decoction, maceration, infusion, digestion, percolation, leaching, hot water entrainment, such as distillation or hydrodistillation, microwave assisted solvent, and ion exchange column . The decoction comprises the following steps: - the boiling of a mixture comprising prickly pear fruits in a polar solvent as defined above; and the maintenance of this boiling for a particular duration, in particular from 1 minute to 2 hours, in particular approximately 20 minutes, then optionally a return to ambient temperature. By "maceration" is meant a method of cold soaking the fruits of prickly pear in the polar solvent for a particular period of time in order to extract the soluble compounds therefrom. In particular, the maceration step has a duration ranging from 1 hour to 15 days, and preferably 1 day. In particular, it is at room temperature, ie 10 to 25 ° C. The infusion is a process of pouring the polar solvent preferably at boiling temperature onto the prickly pears fruits and then allowing the mixture to infuse for a period of one minute to two hours. Digestion is a process of heating a mixture comprising the prickly pear fruit in the polar solvent at a temperature below the boiling point, for example in the case of water at 10 to 89 ° C, and 25 to 70 ° C.
On entend par « percolation » ou « lixiviation », un procédé consistant à faire passer le solvant polaire à travers les fruits de figuier de Barbarie. La distillation peut être effectuée par exemple par VMHD (hydrodistillation par micro-onde sous vide pulsé). Lors de cette étape d'extraction, le solvant polaire est porté à une pression supérieure à la pression atmosphérique. En particulier lorsque le solvant est de l'eau, celle-ci peut être à l'état subcritique, c'est-à-dire que l'eau se trouve à une température comprise entre 100 et 374°C et à une pression entre 1 et 218 bars. Dans ces conditions, l'eau se trouve sous forme liquide. By "percolation" or "leaching" is meant a process of passing the polar solvent through the fruits of prickly pear. The distillation can be carried out for example by VMHD (pulsed vacuum microwave hydrodistillation). During this extraction step, the polar solvent is brought to a pressure greater than atmospheric pressure. In particular when the solvent is water, it may be in the subcritical state, that is to say that the water is at a temperature between 100 and 374 ° C. and at a pressure between 1 and 218 bars. Under these conditions, the water is in liquid form.
En fin d'étape d'extraction, le solvant polaire est porté à une température comprise dans une gamme allant de la température ambiante (25°C) à une température de 300°C au-dessus de la température d'ébullition du solvant utilisé, et de préférence à la température d'ébullition du solvant utilisé. Ainsi, le solvant polaire est au moins en partie réduit, voire totalement éliminé, notamment par évaporation, par sublimation, et de préférence sous pression réduite. On entend par « température d'ébullition du solvant », la température la plus élevée que le solvant peut atteindre avant de s'évaporer, sous forme gazeuse, librement. Cette température se calcule à la pression atmosphérique de 1 atmosphère (101,3 kPa). At the end of the extraction step, the polar solvent is brought to a temperature ranging from ambient temperature (25 ° C.) to a temperature of 300 ° C. above the boiling point of the solvent used. and preferably at the boiling point of the solvent used. Thus, the polar solvent is at least partially reduced or completely eliminated, in particular by evaporation, by sublimation, and preferably under reduced pressure. The term "boiling point of the solvent" means the highest temperature that the solvent can reach before evaporating, in gaseous form, freely. This temperature is calculated at atmospheric pressure of 1 atmosphere (101.3 kPa).
L'étape d'extraction du procédé selon l'invention peut être continue ou discontinue. Le procédé selon l'invention peut comprendre une étape impliquant le chauffage, en particulier d'une durée allant de 1 seconde à 15 jours, et notamment de 10 secondes à 8 heures. The extraction step of the process according to the invention may be continuous or discontinuous. The method according to the invention may comprise a step involving heating, in particular of a duration ranging from 1 second to 15 days, and in particular from 10 seconds to 8 hours.
Lors d'une étape supplémentaire, l'extrait récupéré après extraction peut être stérilisé, notamment par chauffage avec ou sans pression et/ou par passage sur une membrane. Cet extrait de fruit de figuier de Barbarie peut également être désinfecté, notamment par chauffage avec ou sans pression ou encore par l'intermédiaire d'un agent désinfectant. Enfin, cet extrait est généralement séché. Ce séchage peut être effectué par lyophilisation et/ou par atomisation. Ce procédé d'extraction par solvant polaire permet ainsi d'obtenir un extrait de fruit de figuier de Barbarie source de bétalaïnes, quercétine et terpènes. In a further step, the extract recovered after extraction can be sterilized, in particular by heating with or without pressure and / or by passage over a membrane. This prickly pear fruit extract can also be disinfected, in particular by heating with or without pressure or by means of a disinfecting agent. Finally, this extract is usually dried. This drying can be carried out by lyophilization and / or by atomization. This polar solvent extraction process thus makes it possible to obtain a fig tree fruit extract of betalaines, quercetin and terpenes.
Ainsi, ce procédé permet d'obtenir un extrait de figue de Barbarie convenant particulièrement pour la composition anti-oxydante selon l'invention. De plus, la composition anti-oxydante présente en outre la caractéristique suivante : du sélénium à partir de 1 % en poids par rapport au poids de ladite composition, de la superoxyde oxydase (SOD) à partir de 1 % en poids par rapport au poids de ladite composition et de la vitamine C à partir de 10 % en poids par rapport au poids de ladite composition, sont exclus de ladite composition antioxydante En effet, la vitamine C à faible dose permet de protéger l'ADN des radicaux libres et de protéger les hydroxylases indispensables à la production d'énergie. Toutefois, à plus haute dose, tout comme le sélénium et la SOD, elle augmenterait la décondensation de l'ADN avec risque d'aboutir à l'échec de la fécondation chez l'homme comme chez la femme. Avantageusement, la composition comprend en outre de la carnitine ou un de leurs dérivés, et la coenzyme Q10. Le présent demandeur a ensuite montré que lorsqu'on utilise, en sus de l'extrait de figue de Barbarie, des vitamines B6, B9, B12, et du zinc, de la carnitine comme la L-carnitine et le coenzyme Q10, une augmentation substantielle de la motilité et de la production des spermatozoïdes et une baisse du DFI sont obtenues chez l'homme. Par conséquent, une meilleure protection de l'ADN est obtenue. De manière avantageuse, la composition comprend également un acide gras polyinsaturé à longue chaîne en w6, des oligo-proanthocyanidines (OPC), de la zéaxanthine ou un de leurs mélanges. L'ajout d'acides gras polyinsaturés (AGPI) notamment à longue chaîne (oméga 6), et/ou d'OPC, tel que des OPC de pin et de la zéaxanthine (caroténoïde) améliore davantage la proctection de l'ADN. Il s'est avéré que les OPC de pin et la zéaxanthine ont une action sur l'oxygène singulet, acteur privilégié de l'oxydation des protéines des spermatozoïdes. Selon une caractéristique de l'invention, les oligo-proanthocyanidines sont issues de l'extrait de pin et l'acide gras polyinsaturé à longue chaîne en w6 est choisi parmi : l'acide y-linolénique, l'acide di-homo-y-linolénique ou l'acide arachidonique. Préférentiellement, la composition comprend en outre du magnésium sous forme de sel ou de complexe, de la vitamine E, de la cystéine ou un de ses dérivés, de l'arginine ou un des ses dérivés, de la tyrosine ou un de ses dérivés, ou une de leurs combinaisons. Le magnésium jouerait un rôle dans l'énergie du spermatozoïde, il permettrait en outre de lutter contre l'asthénie des spermatozoïdes. La vitamine E permettrait de protéger la membrane lipidique contre la lipopéroxydation et permettrait ainsi d'apporter une stabilisation membranaire. En particulier, la composition comprend un extrait riche en polyphénols choisi parmi : un extrait de romarin, un extrait de thé vert, un extrait de mélisse, un extrait de basilic, un extrait de thym, un extrait de péricarpe et fruit de citrus, un extrait de baies, un extrait d'orthosiphon, un extrait de camu camu, un extrait de raisin, de fruit d'olivier, d'écorce de pin, ou un de leurs mélanges. De préférence, la composition comprend outre la zéaxanthine, d'autres caroténoïdes choisis parmi : l'astaxanthine, le lycopène, la lutéine, le F3- carotène, ou un de leurs mélanges. Selon une autre caractéristique de l'invention, la zéaxanthine issue de l'extrait de tagète, l'astaxanthine est issu de l'extrait d'Hæmatococcus pluvialis, le lycopène est issu de l'extrait de tomate, le F3- carotène est issu d'un extrait de Dunaliella. Le présent demandeur a en effet trouvé que la composition anti-oxydante selon l'invention (extrait de figues de Barbarie, Vit B6,B9, B12 et zinc), associée aux polyphénols des extraits de feuilles de thé vert (Thea sinensis), de fruits d'olivier (Olea europaea), d'écorce de pin (Pinus pinaster), de grenade (Punica granatum), aux flavonoïdes de l'extrait de péricarpes et pulpes de Citrus (Citrus aurantium et Citrus limonium), au lycopène de l'extrait de pulpe de tomate (Solanum lycopersicum) et autres caroténoïdes, tels que l'astaxanthine et la zéaxanthine, permettrait de réduire l'indice de fragmentation de l'ADN ainsi que l'indice de décondensation de l'ADN afin d'améliorer la qualité des spermatozoïdes chez l'homme. Les actifs peuvent également être associés à d'autres choisis parmi : vitamine D et autres vitamines, acide alpha-lipoïque, et autres oligoéléments et minéraux, des acides aminés, glutathion, et autres enzymes. Un autre but de la présente invention concerne un complément alimentaire comprenant la composition anti-oxydante telle que décrite ci-dessus. Un complément alimentaire est par définition une denrée alimentaire dont le but est de compléter un régime normal et qui constitue une source concentrée de nutriments ou d'autres substances ayant un effet nutritionnel ou physiologique, seule ou combinée, commercialisée sous forme de doses. Par dose, on entend toutes les formes classiques : gélules, pastilles, comprimés, ampoules, ou encore sachets. Thus, this method makes it possible to obtain a prickly pear extract that is particularly suitable for the antioxidant composition according to the invention. In addition, the antioxidant composition also has the following characteristic: selenium from 1% by weight relative to the weight of said composition, superoxide oxidase (SOD) from 1% by weight relative to the weight of said composition and vitamin C from 10% by weight relative to the weight of said composition, are excluded from said antioxidant composition In fact, low-dose vitamin C makes it possible to protect the DNA from free radicals and to protect the hydroxylases essential for the production of energy. However, at higher doses, like selenium and SOD, it would increase the decondensation of DNA with the risk of failure of fertilization in both men and women. Advantageously, the composition further comprises carnitine or one of their derivatives, and coenzyme Q10. The present applicant has then shown that when, in addition to the prickly pear extract, vitamins B6, B9, B12, and zinc, carnitine such as L-carnitine and coenzyme Q10 are used, an increase of Substantial motility and sperm production and a decrease in DFI are obtained in humans. Therefore, better protection of the DNA is obtained. Advantageously, the composition also comprises a w6 long-chain polyunsaturated fatty acid, oligo-proanthocyanidines (OPC), zeaxanthin or a mixture thereof. The addition of polyunsaturated fatty acids (PUFAs) including long chain (omega 6) and / or OPC, such as pine OPCs and zeaxanthin (carotenoid) further improves the protection of DNA. It has been shown that pine OPCs and zeaxanthin have an action on singlet oxygen, a key player in the oxidation of spermatozoa proteins. According to one characteristic of the invention, the oligo-proanthocyanidines are derived from pine extract and the long-chain polyunsaturated fatty acid in w6 is chosen from: γ-linolenic acid, di-homo-γ acid -linolenic or arachidonic acid. Preferably, the composition further comprises magnesium in salt or complex form, vitamin E, cysteine or a derivative thereof, arginine or a derivative thereof, tyrosine or a derivative thereof, or one of their combinations. Magnesium plays a role in the energy of spermatozoa, it would also help fight against sperm asthenia. Vitamin E would protect the lipid membrane against lipoperoxidation and thus provide membrane stabilization. In particular, the composition comprises a polyphenol-rich extract selected from: a rosemary extract, a green tea extract, a lemon balm extract, a basil extract, a thyme extract, a pericarp extract and citrus fruit, a berry extract, orthosiphon extract, camu camu extract, grape extract, olive fruit, pine bark, or a mixture thereof. Preferably, the composition comprises, besides zeaxanthin, other carotenoids chosen from: astaxanthin, lycopene, lutein, F3-carotene, or a mixture thereof. According to another characteristic of the invention, the zeaxanthin resulting from the tagete extract, the astaxanthin is derived from the Hæmatococcus pluvialis extract, the lycopene is derived from the tomato extract, the F3-carotene is derived from from an extract of Dunaliella. The present applicant has indeed found that the antioxidant composition according to the invention (extract of prickly pears, Vit B6, B9, B12 and zinc), associated with polyphenols extracts of green tea leaves (Thea sinensis), olives (Olea europaea), pine bark (Pinus pinaster), pomegranate (Punica granatum), flavonoids from the extract of pericarp and Citrus pulp (Citrus aurantium and Citrus limonium), lycopene extract of tomato pulp (Solanum lycopersicum) and other carotenoids, such as astaxanthin and zeaxanthin, would reduce the fragmentation index of DNA as well as the decondensation index of DNA to improve the quality of spermatozoa in humans. The active ingredients may also be associated with others selected from: vitamin D and other vitamins, alpha-lipoic acid, and other trace elements and minerals, amino acids, glutathione, and other enzymes. Another object of the present invention relates to a dietary supplement comprising the antioxidant composition as described above. By definition, a food supplement is a foodstuff whose purpose is to supplement a normal diet and which constitutes a concentrated source of nutrients or other substances having a nutritional or physiological effect, alone or in combination, marketed in the form of doses. By dose is meant all conventional forms: capsules, lozenges, tablets, ampoules, or sachets.
Les différents constituants du complément alimentaire peuvent être administrés simultanément (dans un même comprimé par exemple) ou comme produits de combinaison pour une utilisation séparée ou étalée dans le temps. En particulier, les compléments alimentaires sont formulés pour être administrés par voie orale. Selon une caractéristique de l'invention, les compléments alimentaires se présentent sous forme d'une solution ou d'une suspension aqueuse dans une ampoule en verre, en stick ou en boisson, en flacon compte gouttes, en spray, en capsules molles, ou à l'état sec de comprimé nus ou enrobés, de dragées, de gélules, de capsules, de poudre, de comprimés effervescents, de granules, de sticks, de pastilles. Lorsque les compléments alimentaires se présentent sous forme de capsule molle ou de gélule, l'enveloppe de ces capsules molles ou de ces gélules peut contenir notamment de la gélatine animale telle que la gélatine de poisson, de la glycérine, ou un matériau d'origine végétale tel qu'un dérivé de cellulose ou d'amidon, ou une protéine végétale. Lorsque les compléments alimentaires se présentent sous forme de gélule, de comprimé, ou de granule, le mélange d'actifs peut être fixé sur un support pulvérulent tel que la silice, la cellulose, et la maltodextrine. Les compléments alimentaires peuvent comprendre tout véhicule ou excipient approprié, acceptable du point de vue nutraceutique, ainsi que des additifs conventionnels, connus de l'homme du métier, tels que des agents de charge, des fluidisants, des extraits naturels, des vitamines, des minéraux, des oligoéléments, des acides aminés, des acides gras, des anti-agglomérants, des huiles naturelles, des arômes, des colorants, des acidifiants, des édulcorants etc. The various constituents of the food supplement can be administered simultaneously (in the same tablet for example) or as combination products for separate use or spread over time. In particular, dietary supplements are formulated to be administered orally. According to one characteristic of the invention, the food supplements are in the form of an aqueous solution or suspension in a glass, stick or beverage ampoule, dropper bottle, spray, soft capsule, or in the dry state of naked or coated tablets, dragees, capsules, capsules, powder, effervescent tablets, granules, sticks, lozenges. When the food supplements are in the form of a soft capsule or capsule, the envelope of these soft capsules or capsules may contain in particular animal gelatin such as fish gelatin, glycerin, or a source material plant such as a cellulose or starch derivative, or a vegetable protein. When the food supplements are in capsule, tablet, or granule form, the mixture of active ingredients may be attached to a powdery carrier such as silica, cellulose, and maltodextrin. Dietary supplements may comprise any suitable vehicle or excipient, acceptable from a nutraceutical point of view, as well as conventional additives, known to those skilled in the art, such as fillers, fluidizers, natural extracts, vitamins, minerals, trace elements, amino acids, fatty acids, anti-caking agents, natural oils, flavors, colorants, acidifiers, sweeteners, etc.
Les charges sont avantageusement la cellulose microcristalline, la maltodextrine de pomme de terre ou de maïs. L'épaississant utilisé de préférence est l'amidon de pomme de terre, l'hydroxypropylméthylcellulose, la pectine de citrus, la gomme de guar, de caroube, agar-agar, konjac, les huiles hydrogénées, la cire d'abeille. The fillers are advantageously microcrystalline cellulose, maltodextrin potato or corn. The thickener used preferably is potato starch, hydroxypropyl methylcellulose, citrus pectin, guar gum, carob, agar-agar, konjac, hydrogenated oils, beeswax.
Les fluidisants peuvent être du silicate de magnésium, du stéarate de magnésium, de la silice colloïdale. Les anti-agglomérants sont ceux utilisés usuellement dans l'industrie alimentaire, tels que le stéarate de magnésium, la silice colloïdale. The fluidizers may be magnesium silicate, magnesium stearate, colloidal silica. Anti-caking agents are those usually used in the food industry, such as magnesium stearate, colloidal silica.
Les vitamines sont choisies parmi, entre autres, les vitamines C, E, B6, B1, B2, B3, B5, B8. En tant qu'acidifiant, de l'acide citrique peut être utilisé dans la composition selon la présente invention. The vitamins are selected from, among others, vitamins C, E, B6, B1, B2, B3, B5, B8. As an acid, citric acid may be used in the composition of the present invention.
Les arômes qui peuvent être utilisés sont variés, tels que l'arôme de café, de citron, de pomme, de pêche, de chocolat, de vanille, de fraise. Les édulcorants utilisés sont, entre autres, l'inuline, le xylitol, l'aspartame, le sirop de glucose, le sirop d'de fructo-oligosaccharide, le maltitol en poudre ou en sirop, l'acésulfame de potassium, le fructooligosaccharide, le sucralose, la néohespéridine, les stéviosides et le cyclamate de sodium. Des acides gras peuvent également être ajoutés au complément alimentaire, tels que oméga 3, galactolipides, mais aussi des minéraux : chrome, bore, magnésium, calcium, fer, molybdène et des acides aminés, tels que tryptophane, leucine, arginine et glycine. The aromas that can be used are varied, such as aroma of coffee, lemon, apple, peach, chocolate, vanilla, strawberry. The sweeteners used are, inter alia, inulin, xylitol, aspartame, glucose syrup, fructo-oligosaccharide syrup, maltitol powder or syrup, acesulfame potassium, fructooligosaccharide, sucralose, neohesperidin, steviosides and sodium cyclamate. Fatty acids can also be added to the dietary supplement, such as omega 3, galactolipids, but also minerals: chromium, boron, magnesium, calcium, iron, molybdenum and amino acids, such as tryptophan, leucine, arginine and glycine.
Tous ces composés ne sont nullement limitatifs, des agents acceptables sur les plans pharmaceutique et/ou alimentaire pouvant être ajoutés à la composition selon la présente invention. Préférentiellement, les compléments alimentaires sont de préférence administrés à l'Homme, une à quatre fois par jour, par dose comprenant de 10 mg à 3 g, de préférence de 20 mg à 2 g et de manière encore plus avantageuse de 100 mg à 900 mg du mélange d'actifs et ce, pendant une période de 5 jours à 3 ans. Les compléments alimentaires selon l'invention permettent d'assurer la protection des spermatozoïdes chez l'homme et des ovocytes chez la femme. Les compléments selon l'invention permettent de prévenir les dégâts oxydatifs (sur les membranes, sur l'ADN) sur l'embryon et diminue ainsi le risque de fausses couches. Ils permettent également de prévenir les risques de dégâts vasculaires précoces au niveau de la nidation (empêcher une élévation de l'homocystéine) et ainsi permettre également, par le biais de ce deuxième mécanisme, de diminuer le risque de fausses couches, notamment de fausses couches précoces. Ainsi, lesdits compléments permettent avantageusement de réduire les risques de fausses couches précoces et les risques de malformations embryonnaires, notamment les malformations: - du tube neural; -vésicales; - cardiaques. Ainsi, la présente invention a également pour objet une composition antioxydante telle que décrite ci-dessus ou complément alimentaire tel que décrit ci- dessus, appliqué pour prévenir et/ou de traiter l'infertilité chez l'Homme, assurer une protection des spermatozoïdes chez l'homme, prévenir les fausses couches ou les malformations embryonnaires chez la femme enceinte. Exemples de réalisation non limitatifs Les exemples suivants sont destinés à illustrer l'invention sans aucunement en limiter la portée. Exemple 1: complément alimentaire recommandé pour l'homme lorsque la fragmentation de l'ADN du spermatozoïde est supérieure à 20% ou est associé à l'élévation de la décondensation de la chromatine Le complément alimentaire selon l'invention répond de préférence à la composition pondérale suivante: Pour 2 comprimés de 869,03 mg, par comprimé : Magnésium marin 80 - 300 mg, extrait de baie d'acérola (vitamine C) 5 - 40 mg, extrait de figues de Barbarie 20 - 200 mg, extrait de feuille de thé vert 20 - 70 mg, citrate de zinc 10 - 30 mg, extrait de Dunaliella (bêta-carotène) 10 - 30 mg, extrait de fruit d'olivier 20 - 50 mg , extrait de péricarpe et de fruit de citrus (bioflavonoïdes) 10 - 40 mg, vitamine E 3 - 10 mg, chlorhydrate de pyridoxine (vitamine B6) 0,5 - 1,5 mg, acide folique (vitamine B9) 0,05 -1 mg, cyanocobalamine (vitamine B12) 0,00030 - 0,000120 mg, extrait de tomate (lycopène) 0,5 - 1,5 mg. All these compounds are in no way limiting, pharmaceutically and / or food acceptable agents that can be added to the composition according to the present invention. Preferably, the food supplements are preferably administered to humans, one to four times per day, per dose comprising from 10 mg to 3 g, preferably from 20 mg to 2 g and even more advantageously from 100 mg to 900 mg. mg of the mixture of assets for a period of 5 days to 3 years. The dietary supplements according to the invention make it possible to protect spermatozoa in humans and oocytes in women. The supplements according to the invention make it possible to prevent oxidative damage (on membranes, on DNA) on the embryo and thus reduce the risk of miscarriages. They also prevent the risk of early vascular damage at implantation (prevent elevation of homocysteine) and thus also allow, through this second mechanism, to reduce the risk of miscarriages, including miscarriages early. Thus, said supplements advantageously make it possible to reduce the risk of early miscarriages and the risk of embryonic malformations, in particular malformations: of the neural tube; -vésicales; - heart. Thus, the subject of the present invention is also an antioxidant composition as described above or a dietary supplement as described above, applied to prevent and / or treat infertility in humans, to ensure protection of spermatozoa in man, prevent miscarriages or embryonic malformations in pregnant women. Nonlimiting examples of embodiment The following examples are intended to illustrate the invention without in any way limiting its scope. Example 1: dietary supplement recommended for humans when the fragmentation of the sperm DNA is greater than 20% or is associated with the elevation of the chromatin decondensation The dietary supplement according to the invention preferably corresponds to the composition weight: For 2 tablets of 869.03 mg, per tablet: Marine Magnesium 80 - 300 mg, Acerola Berry Extract (Vitamin C) 5 - 40 mg, Prickly Pear Extract 20 - 200 mg, Leaf Extract of green tea 20 - 70 mg, zinc citrate 10 - 30 mg, Dunaliella extract (beta-carotene) 10 - 30 mg, olive fruit extract 20 - 50 mg, extract of pericarp and citrus fruit (bioflavonoids ) 10 - 40 mg, vitamin E 3 - 10 mg, pyridoxine hydrochloride (vitamin B6) 0.5 - 1.5 mg, folic acid (vitamin B9) 0.05 - 1 mg, cyanocobalamin (vitamin B12) 0.00030 - 0.000120 mg, tomato extract (lycopene) 0.5 - 1.5 mg.
Pour ce comprimé particulier, la dose recommandée est de deux comprimés par jour. Exemple 2 : complément alimentaire recommandé pour l'homme lorsque la fragmentation de l'ADN du spermatozoïde est supérieure à 40% Le complément alimentaire selon l'invention répond de préférence à la composition pondérale suivante: Pour 1 comprimé de 850 mg, par comprimé : L-cystéine 50 - 250 mg, extrait de figues de Barbarie 50 - 200 mg, L-carnitine 40 - 150 mg, L-tyrosine 40 - 150 mg, extrait de tagète (zéaxanthine) 60 - 120 mg, extrait d'Hæmatococcus pluvialis (astaxanthine) 30 - 90 mg, extrait de feuille de thé vert (polyphénols) 40 - 80 mg, citrate de zinc 10 - 30 mg, bêta-carotène 2 - 5 mg, extrait de fruit d'olivier 25 - 50 mg, extrait de baie d'acérola (vitamine C) 10 - 50 mg, extrait de péricarpe et de fruit de citrus (bioflavonoïdes) 10 - 30 mg, coenzyme Q10 10 - 40 mg, extrait d'écorce de pin maritime (OPC) 10 - 50 mg, vitamine E 3 - 10 mg, chlorhydrate de pyridoxine (vitamine B6) 1 - 3 mg, extrait de tomate (lycopène) 0,5 - 2 mg, acide folique (vitamine B9) 0,04 -1 mg, cyanocobalamine (vitamine B12) 0,00020 - 0,000120 mg, Pour une capsule de 500 mg, par capsule : huile de foie de requin (alkylglycérols) 200 - 500 mg. Pour ce comprimé et cette capsule particuliers, la dose recommandée est de un comprimé et une capsule par jour. For this particular tablet, the recommended dose is two tablets a day. Example 2: dietary supplement recommended for humans when the fragmentation of the sperm DNA is greater than 40% The dietary supplement according to the invention preferably corresponds to the following weight composition: For 1 tablet of 850 mg per tablet: L-cysteine 50 - 250 mg, prickly pear extract 50 - 200 mg, L-carnitine 40 - 150 mg, L-tyrosine 40 - 150 mg, tagete extract (zeaxanthin) 60 - 120 mg, Haematococcus pluvialis extract (astaxanthin) 30 - 90 mg, green tea leaf extract (polyphenols) 40 - 80 mg, zinc citrate 10 - 30 mg, beta-carotene 2 - 5 mg, olive fruit extract 25 - 50 mg, extract acerola berry (vitamin C) 10 - 50 mg, extract of pericarp and citrus fruit (bioflavonoids) 10 - 30 mg, coenzyme Q10 10 - 40 mg, extract of maritime pine bark (OPC) 10 - 50 mg, vitamin E 3 - 10 mg, pyridoxine hydrochloride (vitamin B6) 1 - 3 mg, tomato extract (lycopene) 0.5 - 2 mg, folic acid (vi B9) 0.04-1 mg, cyanocobalamin (vitamin B12) 0.00020 - 0.000120 mg, For a 500 mg capsule, per capsule: shark liver oil (alkylglycerols) 200 - 500 mg. For this particular tablet and capsule, the recommended dose is one tablet and one capsule per day.
Exemple 3: complément alimentaire recommandé pour l'homme lorsque la décondensation de l'ADN est isolée Le complément alimentaire selon l'invention répond de préférence à la composition pondérale suivante: Pour 1 comprimé de 500 mg, par comprimé : extrait de figues de Barbarie 50 - 300 mg, citrate de zinc 10 - 30 mg, chlorhydrate de pyridoxine (vitamine B6) 0,5 - 1,5 mg, acide folique (vitamine B9) 0,05 -1 mg, cyanocobalamine (vitamine B12) 0,00030 - 0,000120 mg. Pour ce comprimé particulier, la dose recommandée est de un comprimé par jour. Example 3: dietary supplement recommended for humans when the decondensation of the DNA is isolated The dietary supplement according to the invention preferably corresponds to the following composition by weight: For 1 tablet of 500 mg per tablet: extract of prickly pears 50 - 300 mg, zinc citrate 10 - 30 mg, pyridoxine hydrochloride (vitamin B6) 0.5 - 1.5 mg, folic acid (vitamin B9) 0.05 - 1 mg, cyanocobalamin (vitamin B12) 0.00030 - 0.000120 mg. For this particular tablet, the recommended dose is one tablet per day.
Exemple 4: complément alimentaire recommandé pour l'homme en cas d'asthénie des spermatozoïdes (moins de 40% de spermatozoïdes mobiles) Le complément alimentaire selon l'invention répond de préférence à la composition pondérale suivante: Pour 2 comprimés de 800 mg, par comprimé : Magnésium marin 80 - 300 mg, extrait de figues de Barbarie 20 - 200 mg, extrait de grenade fruit 20 - 200 mg, citrate de zinc 10 - 30 mg, vitamine E 3 - 10 mg, chlorhydrate de pyridoxine (vitamine B6) 0,5 - 2 mg, acide folique (vitamine B9) 0,05 -1 mg, cyanocobalamine (vitamine B12) 0,00030 - 0,000120 mg, N-acétyl cystéine 20 - 200 mg, L-arginine 50 - 500 mg, L-carnitine 70 - 140 mg, coenzyme Q10 10 - 30 mg. Pour ce comprimé particulier, la dose recommandée est de deux comprimés par jour. Exemple 5 : complément alimentaire recommandé pour la femme en péri conception Le complément alimentaire selon l'invention répond de préférence à la composition pondérale suivante: Pour 1 comprimé de 800 mg: extrait de figues de 10 Barbarie 50 - 250 mg, gluconate de zinc 10 - 30 mg, chlorhydrate de pyridoxine (vitamine B6) 1 - 3 mg, acide folique (vitamine B9) 0,05 - 1 mg, cyanocobalamine (vitamine B12) 0,00030 - 0,000120 mg. Pour ce comprimé particulier, la dose recommandée est de un comprimé par Bien que l'invention ait été décrite en relation avec un mode de réalisation particulier, il est bien évident qu'elle n'y est nullement limitée et qu'elle comprend tous les équivalents techniques des moyens décrits ainsi que leurs combinaisons si celles-ci entrent dans le cadre de l'invention. jour. 15 20 Example 4: dietary supplement recommended for humans in the event of asthenia of spermatozoa (less than 40% of motile spermatozoa) The dietary supplement according to the invention preferably corresponds to the following weight composition: For 2 tablets of 800 mg, per tablet: Marine Magnesium 80 - 300 mg, Prickly Pear Extract 20 - 200 mg, Pomegranate Fruit Extract 20 - 200 mg, Zinc Citrate 10 - 30 mg, Vitamin E 3 - 10 mg, Pyridoxine Hydrochloride (Vitamin B6) 0.5 - 2 mg, folic acid (vitamin B9) 0.05 -1 mg, cyanocobalamin (vitamin B12) 0.00030 - 0.000120 mg, N-acetyl cysteine 20 - 200 mg, L-arginine 50 - 500 mg , L-carnitine 70 - 140 mg, coenzyme Q10 10 - 30 mg. For this particular tablet, the recommended dose is two tablets a day. Example 5: dietary supplement recommended for the woman during conception The dietary supplement according to the invention preferably corresponds to the following weight composition: For 1 tablet of 800 mg: extract of prickly pear 50 - 250 mg, zinc gluconate 10 - 30 mg, pyridoxine hydrochloride (vitamin B6) 1 - 3 mg, folic acid (vitamin B9) 0.05 - 1 mg, cyanocobalamin (vitamin B12) 0.00030 - 0.000120 mg. For this particular tablet, the recommended dosage is one tablet per tablet. Although the invention has been described in connection with a particular embodiment, it is quite obvious that it is in no way limited and that it includes all technical equivalents of the means described and their combinations if they fall within the scope of the invention. day. 15 20
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