• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The present invention relates to the use of a composition comprising Norbixinobtained by purification from a seed extract of Bixa orellana, for the photoprotection of the cells of the retinal pigment epithelium (RPE) in the mammal.
    公开号:FR3035589A1
    申请号:FR1553957
    申请日:2015-04-30
    公开日:2016-11-04
    发明作者:Rene Lafont；Stanislas Veillet；Jose-Alain Sahel；Valerie Fontaine；Pierre-Paul Elena
    申请人:Universite Pierre et Marie Curie Paris 6；Institut Biophytis SAS；Iris Pharma；
    IPC主号:
    专利说明:

    [0001] FIELD OF THE INVENTION The present invention relates to the field of treatments for retinal pigment epithelium (EPR) cells. More particularly, the present invention relates to the use of a composition for the protection of retinal pigment epithelium (RPE) cells, in particular for the treatment of age-related macular degeneration (AMD) or of the disease Stargardt and retinitis pigmentosa in mammals. The object of the invention is to improve the vision of individuals suffering from these diseases or at least to stabilize the course of the disease.
    [0002] BACKGROUND Age-related macular degeneration (AMD) is a cause of irreversible blindness in the elderly population, particularly in Europe and North America. AMD affects the central part of the retina, called macula, causing severe visual impairment and irreversible loss of central vision.
    [0003] Macular function is at the origin of central vision and visual acuity, the high resolution of which is related to its high concentration of cone photoreceptors. The early stage of AMD is marked by deposits called Drüsen, which affect vision only marginally. Subsequent phases include two forms of AMD, geographic atrophy (dry form) or exudative form (wet or neovascular zo), the former being much more common than the second. The latter stages of both forms lead to the destruction of the macular neurosensory retina, but the course of dry AMD is generally slow, whereas wet AMD can lead to complete blindness within a few weeks. Aging is the gradual accumulation over time of changes that are associated with (or responsible for) increasing susceptibility of the disease. In the retina, a number of degenerative diseases, including glaucoma, retinitis pigmentosa, and AMD, may occur as a result of aging. Retinitis pigmentosa is a heterogeneous group of 30 retinal degenerations, involving photoreceptors and EPR, leading to a loss of night vision, and later to central vision. Although the specific mechanisms involved in the initiation of different types of retinal aging-related diseases differ, it is believed that the resulting oxidative stress and inflammation are important components that contribute to the pathogenesis. Theories of the etiology of AMD include hydrodynamic changes in the Bruch membrane caused by progressive accumulation of lipid-containing extracellular material, and EPR senescence, the activity of which is essential for photoreceptor survival. . EPR cells have several different functions in the eyes: they establish the blood-retinal barrier by their tight junctions, and thus are responsible for the immuno-privileged status of the inner part of the eye bulb; they keep photoreceptors alive by providing nutrients and participating in the visual cycle. The current understanding is that a deficiency in the function of EPR cells is at the root of the development of AMD. Aging causes EPR cell dysfunction and poor metabolism, as well as phagocytic activity. Incomplete digestion of the outer segments of the photoreceptors can lead to Drüsen formation by reducing diffusion across the Bruch membrane, initially causing deformation of the retina and perceived images.
    [0004] With age, EPR stores an increasing amount of lipofuscins. These are composed of lipids and proteins, which come from phagolysosomes, lysosomes and photoreceptors. Lipofuscins also contain N-retinyl-N-retinylidene ethanolamine (A2E), which is formed by the condensation of two molecules of retinaldehyde with ethanolamine.
    [0005] Aging is accompanied by increased accumulation of A2E in the retina (Bhosale et al., 2009). Under the action of blue light and in the presence of oxygen, A2E generates reactive species that cause damage to proteins, lipids and DNA, and therefore significant oxidative stress in the aging cells of the body. EPR (Sparrow & Cai, 2001). This damage disrupts the lysosomal activity of EPR cells and causes an accumulation of waste, which eventually leads, from place to place, to the death of EPR cells, which is followed by that of the photoreceptors with which they were associated. . No drug exists on the market for the treatment of dry AMD, while intravitreal injection of anti-VEGF 303 5 5 8 9 3 (Vascular Endothelial Growth Factor) drugs are commercially available to partially block the formation of neovessels and thus offering an alternative treatment for wet AMD. Food supplements have been formulated with generic antioxidant compounds, namely vitamins and minerals with antioxidant properties, for example zinc, vitamins A, C, E, with actual but limited therapeutic efficacy. The AREDS nutraceutical 1 (Age-Related Eye Disease Study, AREDS 2001) is considered the standard of care in the United States for the treatment of dry AMD, reducing the risk of advanced AMD by 25% and the vision loss of 19% over five years.
    [0006] Many products offer a common formulation base: Zinc and vitamins C and E, to which are added various ingredients: lutein, resveratrol, Omega 3 fatty acids, without however having convincing efficacy data on these additional ingredients, or on the categories of patients likely to respond favorably to these different molecules (Elliot & Williams, 2012).
    [0007] Carotenoids (molecules exclusively provided by the diet) have been more particularly studied because some of them (lutein, zeaxanthin = xanthophylls) are naturally present in the macula (Subczynski et al., 2010), and these compounds are known to possess a high antioxidant capacity. It is therefore logically that these compounds have been tested (alone or in combination) in the AREDS formula, but the results obtained have been limited, the supplementation being effective only for a subset of patients deficient in these compounds. (Pinazo-Durán et al., 2014). These molecules are effective in vitro to protect EPR cells (Human D407) against the toxic effects of hydrogen peroxide (Pintea et al., 2011).
    [0008] Other xanthophylls have also been studied by oral supplementation alone or in combination with lutein and / or zeaxanthin (eg astaxanthin - Parisi et al., 2008). Recently, diapocarotenoids (= carotenoids truncated at both ends - IUPAC chemical nomenclature) have been tested in vitro and in vivo, in particular crocetin (= 8,8'-diapocarotene-8,8'-dioate) and its glycosides ( the crocines). Crocins have an in vitro photoprotective effect on primary cultures of cattle or primate photoreceptors (Laabich et al., 2006), and crocetin protects neuroganglion cells against oxidative stress (Yamauchi et al., 2011). Saffran (a crocin / crocetin-rich spice) administered orally has been shown to be active in vivo on retinal quality (Maccarone et al., 2008; Falsini et al., 2010; Boisti et al., 2014). ). However, saffron containing other molecules that may be active on the retina, like other carotenoids as well as saffron (Verma & Middha, 2010, Fernandez Sanchez et al., 2012), it is difficult to conclude as to the effect the only crocetin.
    [0009] Experiments were also carried out with another apocarotenoid, bixin (= methylhydrogen 6,6'-diapocarotene-6,6'-dioate) or some of its derivatives, in vitro on neuroganglion cells and in vivo by intravitreal injection for counteract the effects of endoplasmic reticulum stress (Tsuruma et al., 2012). The tests thus carried out most often evaluate antioxidant and therefore protective activity of the compounds with respect to various retinal cell types subjected to the presence of an oxidizing agent (eg hydrogen peroxide), and they are therefore not directly in the context of AMD. A previously developed Urucum seed extract (Bixaorellana) (Bixilia®) has shown a photoprotective effect on human skin exposed to UV (Veillet et al., 2009) and on EPR cells under photo-oxidative stress ( Fontaine et al., 2011). Bixilia® extract is a natural extract of Urucum that has been enriched with Bixin. Bixilia® contains other photoprotective compounds of phenolic nature, the presence of which could explain the higher photoprotective activity of the crude extract compared to Bixine alone. In patent FR 11 54172 (Fontaine et al., 2011), the protective effect of the EPR cells of some of the compounds of the Bixilia® extract is analyzed. The results of tests using Bixin or Norbixin at 0.1 micromolar (pM), 1 μM and 10 μM did not have photoprotective activity and suggested that the higher the concentration of Bixin or Norbixin, the lower the concentration. EPR cells survive and therefore less the photoprotective effect is great. Among others, it is noted that substances such as cyanidin and ellagic acid at concentrations of 10 μM and 20 μM have a beneficial photoprotective effect on EPR cells. An in-depth study has led to identifying the active molecules present in the Bixilia® extract and to clarifying their mechanism of action, and then to demonstrating their efficacy in vivo in mice and rats. This study gave birth to the present invention. The invention thus provides alternative treatment to those already existing for the protection of EPR cells.
    [0010] OBJECT OF THE INVENTION The inventors have discovered that Norbixin, in particular its 9'-cis form, makes it possible to greatly reduce cell death caused by illumination with blue rays of EPR cells pretreated with N-Retinyl-N-Retinylidene Ethanolamine (A2E). According to a first aspect, the present invention is directed to a composition containing norbixin obtained by purification from a seed extract of Bixa orellana, for its use for the photoprotection of retinal pigment epithelium (RPE) cells in mammals .
    [0011] In the context of the invention, the term "Bixa orellana seed extract" means an extract prepared from the outer part of the seeds, that is to say the waxy substance covering the seeds of Bixa orellana. This waxy substance is known to be rich in bixin and other minor carotenoids, as well as for its use as a food coloring agent.
    [0012] Norbixin, bioavailable in mammals after oral administration, is much better absorbed than Bixin and is found in particular in the retina. In particular embodiments of the invention, the composition comprises more than 90% by weight of Norbixin.
    [0013] In particular embodiments of the invention, the composition comprises more than 90% by weight of Norbixin in its 9'-cis form of formula (I): CH 3 CH 3 (I) In particular embodiments, the composition comprises at least one element selected from zinc, vitamin C and vitamin E.
    [0014] In particular embodiments, the composition may be used in the form of a dietary supplement or a drug. "Food supplement" means a product containing said composition for the purpose of supplementing the diet by providing nutrients beneficial to health as defined by the European Directive 2002/46 / EC. For example, a dietary supplement may be a capsule or tablet to be swallowed or a powder or small ampoule to be mixed with a food and having beneficial effects on the EPR cells. By drug is meant a product containing a precise dose of said compound 5 or said extract according to the definition given by the European Directive 65/65 / EC, ie any substance or composition presented as possessing curative or preventive properties with regard to human diseases or animal. For example, the drug containing the compound at therapeutic doses can be administered orally in capsule or tablet form or injected intravitreously or any other route to confer beneficial effects on the retina. In particular embodiments, the composition comprises a carrier acceptable to be ingested, injected into the eye, injected systemically or injected into the blood. In embodiments, the composition is administered to the mammal per day in an amount of from 0.48 mg / kg of body weight to 48 mg / kg of body weight, preferably in the range of 0.6 mg / kg of body weight. body weight and 20 mg / kg body weight. According to other particular embodiments of the invention, the composition is intended to prevent damage to the retina that may be caused by blue light exposure. By blue radiation is meant radiation corresponding to the blue band of the spectrum of visible light, wavelength between 435 nm and 490 nm. In particular embodiments of the invention, the composition is for the treatment of age-related macular degeneration (AMD) in the mammal. In other particular embodiments, the composition is for treating Stargardt's disease and / or retinitis pigmentosa in the mammal. Stargardt's disease, or Stargardt's syndrome, is an inherited condition, associating a decrease in bilateral visual acuity with atrophy of the macula, which reproduces, at an early age, the symptoms of the dry form of AMD. Brief Description of the Figures Figure 1 illustrates the percentage of surviving EPR cells in the presence of N-retinyl-N-retinylidene ethanolamine (A2E) and Bixilia® or Bixine extract (20 μM) or Norbixin ( 20 i_iM) after being subjected to illumination. Figure 2 illustrates the photoprotective activity of the successive extracts of the seeds of Urucum (C = cyclohexane, D = dichloromethane, M = methanol) on the EPR cells placed in the presence of A2E and subjected to illumination. Figure 3a illustrates the plasma concentrations after ingestion of Bixin (left) or Norbixin (right) in C57BI / 6 mice. Figure 3b illustrates the pharmacokinetic analysis of Norbixin in the C57BI / 6 mouse. Figure 4 illustrates the HPLC-MS / MS analysis of Norbixin in the eyes of double KO mice (ABCA4 - / -, RDH8 - / -) after intraperitoneal injection (10 mg / kg). (3: injected Norbixin, 1 and 2: mono-glucuronides of this compound). Figure 5 illustrates the kinetics of A2E accumulation in the double KO mouse eye (ABCA4 - / -, RDH8 - / -) as a function of age, compared with normal mice. Figure 6 illustrates the electroretinograms (left-hand A-wave and right-hand-right-B-wave) of double KO mice (ABCA4 - / -, RDH8 - / -) that received unilateral intravitreal injections of Norbixin (in order to obtain a final concentration in 20 the vitreous 130 pM), placed for 24 hours in the dark and then exposed to blue light (4000 lux, 1 h). Electroretinograms are performed 7 days after illumination. Figure 7 illustrates the number of photoreceptor core layers as a function of distance from the optic nerve in the eyes of treated mice as in Figure 6. Figure 8A illustrates the experimental protocol of creating the rat blue model. light ". Figure 8B illustrates the results of the electroretinograms of the rats injected with norbixin (100 mg / kg, 4 injections per rat of a 50 mM solution in 9% NaCl, 4 rats / series). using PBN (phenyl-N-tert-butylnitrone, 50 mg / kg, 20 mg / mL solution in 9% NaCl) as a positive control. Electroretinograms are performed 7 days after treatment. Figure 9A illustrates the number of photoreceptor core layers as a function of distance from the optic nerve in rat eyes following intraperitoneal injections of alpha-phenyl-N-tert-butyl nitrone (PBN) or Norbixin and illumination. with a blue light. Histological analyzes are performed 7 days after treatment. Figure 9B illustrates the area under each curve of Figure 9A.
    [0015] Figure 10 illustrates the amount of A2E accumulated in the eye of a double KO mouse (ABCA4 - / -, RDH8 - / -) having or not ingested food supplemented with norbixin for 3 months. Figure 11 illustrates the electroretinograms of double KO mice (ABCA4 - / -, RDH8 - / -) fed or unsupplied with food containing 0.3 mg / g of pure norbixin for 3 months. Figure 12 illustrates the relationship between the amplitude of the electroretinogram (A wave) and the amount of A2E accumulated in the eyes of the double KO mice (ABCA4 - / -, RDH8 - / -). Figure 13 illustrates the results of reverse phase HPLC analysis of norbixin purified from Bixa orellana extract (isomer identification was performed according to Scotter et al., 1998 and Polar-Cabrera et al. al., 2010). DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION Unlike most previously published studies, the models used in the present invention (both in vitro and in vivo) emphasize the role of N-retinyl. -N-retinylidene ethanolamine (A2E) and its phototoxicity, and are therefore closer to human pathology. The tests used in vitro are similar in principle to those used with other natural substances on a human EPR cell line (ARPE-19 cells - Young et al., 2005). Protocols and Results 1- Preparation of Bixin and Norbixin Bixine 95% by weight pure is prepared from a commercial product (Annatto B) from an organic extraction of Urucum seeds and a concentration of Bixin greater than 85% by weight. The purification is carried out by successive recrystallizations. 95% by weight pure Norbixin is obtained after alkaline hydrolysis of the purified Bixin (5% KOH, 60 ° C., 3 hours). The resulting solution is acidified with concentrated hydrochloric acid and Norbixin is recovered by centrifugation. The pellet is washed twice with water to remove the salts, and the final pellet is lyophilized. The purity of the compounds is assessed by UV-Vis spectrophotometry and reversed-phase HPLC: the compounds essentially contain the 9'-cis isomers (concentration greater than 90% by weight, FIG. 13). 2- In vitro tests An in vitro test to study the photoprotective effect of various natural substances on EPR cells placed in the presence of A2E was used. The photoprotective effect of the molecules is evaluated in a cellular model of phototoxicity induced by A2E treatment followed by illumination in blue light. Blue radiation is understood to mean the radiation corresponding to the blue band of the visible light spectrum, ie of wavelength between 435 and 490 nm. This model uses primary cultures of adult pig EPR. Cell survival is quantified through a cell viability test. The test compounds (in 5 mM solution in DMSO) are added at -48h to obtain final concentrations of 1 to 20 μM) and then at -19h of the A2E (final concentration 25 μM) and the cells are irradiated ( time Oh). 24h is measured after the survival of the cells. Image acquisition and processing are performed using a fluorescence microscope driven by the Metamorph software and a dedicated quantification program. The experiments are carried out on 96 well microplates in quadruplicate and the experiment is reproduced at least four times. The results are expressed as a ratio representing the number of living cells in the wells treated with the test molecules, divided by the number of living cells in the control wells (treated with the dilution medium without A2E).
    [0016] This test has previously made it possible to demonstrate the very good photoprotective activity of an ethanolic extract of Annatto seeds (Bixilia® - see Fontaine et al., 2011). In the previous work, if the activity of the Annatto extract had been demonstrated, the nature of the photoprotective substance (s) had not been identified, and the main component of this The extract (bixin) was ineffective at concentrations of 0.1 μM, 1 μM and 10 μM. Additional work has therefore been undertaken to identify the active compound (s). at. Bixine is responsible for much of the photoprotective activity of Bixilia®. Figure 1 shows that Bixin and Norbixin (20 μM) effectively protect EPR cells from A2E induced phototoxicity by compared to the control with A2E. A crude extract of Urucum seeds diluted to provide 20 μM bixin has a high photoprotective activity. The use of very pure bixin at a concentration of 20 μM made it possible to show that this component actually possessed a significant photoprotective activity (FIG. 1) and that this explained a significant part of the activity of the extract. Bixilia® diluted to provide the same amount of bixin. Similar activity has also been found for Norbixin, which is the major circulating metabolite of Bixine (Levy et al., 1997). These results are in agreement with the photoprotective activity of these same compounds, previously demonstrated for the photoprotection of human skin (Veillet et al., 2009). b. Bixilie contains other photoprotective compounds Bixilia® contains other phenolic photoprotective compounds, the presence of which could explain the higher activity of the crude extract compared to bixin alone (for the same concentration of bixin). Sequential extraction of the envelope of the seeds of Urucum was carried out successively with cyclohexane, dichloromethane and methanol (1 L of each / 100 g of seeds).
    [0017] After extraction with cyclohexane, a fraction with a Bixin concentration of 0.65 μM is obtained, after extraction with dichloromethane a fraction is obtained which has a Bixin concentration of 1485 μM, and after extraction with methanol a fraction which has a concentration of Bixine of 4511M.
    [0018] The previous in vitro test is then reproduced. According to FIG. 2, the dichloromethane fraction, which contains 97% of Bixin, is very active, but it is also noted that the methanolic extract rich in phenolic compounds has a significant activity (C = cyclohexane, D = 5 dichloromethane, M = methanol). ). 3- Bioavailability of Bixin and Norbixin Studies of the bioavailability of Bixin and Norbixin were performed in C57BI / 6 mice. The compounds were administered orally (50 mg / kg). Blood samples were taken after 0.25, 0.5, 1, 3, 6, 8 and 24 h and analyzed by HPLC-DAD (UV 460 nm) -MS / MS. Table 1 and Figure 3a reveal that the ingested Bixin is rapidly converted to Norbixin and that the two compounds circulate at comparable concentrations and are not detected after 8 hours. Ingested Norbixin is also significantly more bioavailable than Bixin. Table 1: Ingestion of bixin (50 mg / kg) Ingestion of norbixin (50 mg / kg) Tmax (h) Cmax AUC 0-24h Tmax (h) Cmax AUC (ng / m L) (ng.h / mL) ( ng / m L) (ng.h / ml) Bixin 0.5 833 1666 Norbixin 0.5 12400 41609 Norbixin 0.5 649 1343 Comparison of plasma analyzes (FIG. 3b) after intraperitoneal injection (5 mg / kg) and oral administration (50 mg / kg) shows that the bioavailability of Norbixin is 55%. The presence of Norbixin in the eyes was investigated in double KO mice (ABCA4 - / -, RDH8 - / -) 3 hours after intraperitoneal injection of Norbixin (10 mg / kg). The eyes of 6 animals were dissected and the samples were extracted with acetonitrile, pooled and analyzed by HPLC-MS / MS (Figure 4), which specifically detected the presence of Norbixin in the patient. EPR and retina (Table 2).
    [0019] 303 5 5 8 9 12 Table 2: Sample Norbixin (ng / organ) EPR 5.15 Retina 2.40 Crystalline <LOQ Mood glazed <LOQ Total 7.55 According to Figure 4, it is noted that in plasma, but also in the eyes, Norbixin is also present in conjugated form: the initial compound gives indeed two mono-glucuronides which are eluted before the original compound and exhibit similar fragmentation, probably due to the decomposition of glucuronides in the spectrometer source massive. Glucuronidation has also been described in the case of crocetin (Asai et al., 2005).
    [0020] It is also possible to observe a cis-trans isomerization of Norbixin, the importance of which varies according to the duration of the experiments. This is a classic phenomenon in (poly) -unsaturated compounds, which corresponds to cis-trans isomerizations of one or more double bonds and has been observed in humans in the case of norbixin by Levy et al. (1997). The compound used here is purified from commercial compounds (Annatto B); it contains most of the 9-cis form and very small quantities of the all-trans form and other cis or di-cis forms (Figure 13). 4- Photoprotective Activity by Intravitreal Injection in Mice A genetically modified mouse model developed by Maeda et al. (2008) was used to test the photoprotective activity of Norbixin. In this mouse model, two genes involved in the visual pigment cycle (ABCA4 and RDH8) are inactivated, resulting in an early accumulation of A2E in the eyes (Figure 5). This animal model is thus related to human pathology, 25 with of course its limits, related to the differences in the organization of eyes between rodents and primates. 7 week old mice were therefore used to perform unilateral intravitreal injections of Norbixin (in order to obtain a final concentration in the 130 pM vitreous). After 24 h in the dark, the mice were exposed to blue light (4000 lux, 1 h). The electroretinograms performed 7 days later showed a protective effect of Norbixin, the presence of which has made it possible to maintain a significant electrical activity as explained in FIG. 6. A histological study of the thickness of the layer of external cores evidence of the protective effect of Norbixin on photoreceptors (Figure 7). It should be noted that Norbixin was virtually eliminated 24 hours after intravitreal injection and is therefore only very weakly present in the eyes at the moment of illumination. 5- Photoprotective activity by systemic (intraperitoneal) injection in rats The "rat blue light" model consists in subjecting the animals to intense blue light for 6 hours in order to cause eye damage which is appreciated 7 days later by the realization of electroretinograms then by histological analyzes. An antioxidant compound, PBN (phenyl-N-tertbutylnitrone) is used as a positive control (Ranchon et al., 2001, Tomita et al., 2005). The compounds whose photoprotective activity is to be determined are injected (intraperitoneally) before and during the illumination phase. This is done with Philips blue neon tubes (4.2 mW / cm2) for 6 hours. The experimental protocol is shown in Figure 8A. Three series of experiments were performed with Norbixin (100 mg / kg, 4 injections per rat of a 50 mM solution in 9% NaCl, 4 rats / series) using PBN (phenyl-N-). tert-butylnitrone, 50 mg / kg, 20 mg / mL solution in 9% NaCl) as a positive control. The analysis of electroretinograms (A and B waves) is presented in Figure 8B. This test has demonstrated a significant efficacy of Norbixin, which is close to that of PBN. The corresponding histological data (FIGS. 9A and 9B) confirm the photoprotective action of Norbixin on the photoreceptor survival. 6- Light-sensitive activity by chronic oral administration in mice 3035589 14 A feed containing 0.3 mg / g of pure Norbixin was prepared and given to double knockout mice (ABCA4 - / -, RDH8 - / -) for a period of 3 months. Animals receiving food supplemented with Norbixin show a reduction in A2E accumulation in the eyes (Figure 10): the difference between the two groups is very significant (p = 0.0109). Food supplemented with Norbixin also has a positive effect on the amplitude of the electroretinogram (ERG) (Figure 11). These analyzes also showed that there is an inverse relationship between the amount of A2E accumulated in the eyes and the amplitude of the ERG (Figure 12), which confirms the role of A2E accumulation in the development of pathology (Wu et al., 2014), and the interest of molecules whose administration reduces the accumulation of A2E in the eyes. However, there is no significant accumulation of Norbixin in the eyes during this chronic treatment, which leads to the conclusion that, unlike xanthophylls, this molecule would be degraded. The non-accumulation of this active substance may be considered an advantage, as the accumulation of some carotenoids (eg canthaxanthin) is likely to lead to the formation of deposits within EPR cells (Goralczyk et al. 1997). It is also indicative of a modifying action of EPR cell activity rather than a filtering or antioxidant role, as postulated for lutein and zeaxanthin. This result matches that noted during intravitreal injections (ie the disappearance of Norbixin at the moment of illumination). The daily dose for significantly slowing retinal degeneration in mice after oral administration is 48 mg / kg body weight. Transposition to humans leads to an active daily dose of 4.8 mg / kg. Admissible daily intake or ADI (Adaptive Daily Intake or ADI) of Norbixin is also known to be at most 0.6 mg / kg / day body weight (JECFA / 67 / FC). This value was established on the basis of a no-effect dose or NOAEL in rats of 69 mg / kg / day body weight, equivalent to a daily no-effect dose. in humans at 11 mg / kg, knowing that no toxicity was observed up to 20 mg / kg / day (Hagiwara et al., 2003). The proposed dosage is between 0.48 mg / kg / day and 48 mg / kg / day, ideally between 0.6 mg / kg / day and 20 mg / kg / day.
    [0021] 303 5 5 8 9 References AREDS Report No. 8. 2001. A randomized, placebo-controlled, clinical trial of highdose supplementation with vitamins C and E, beta carotene, and zinc for age-related macular degeneration and vision loss. Arch Ophthalmol, 119: 1417-1436. Asai A, Nakano T, Takahashi M, Nagao A. 2005. Orally administered crocetin and crocins are absorbed into blood plasma as crocetin and its glucuronide conjugates in mice. J Agric Food Chem, 53: 7302-7306. Bhosale P, Serban B, Bernstein PS. 2009. Retinal carotenoids can attenuate 10 formation of A2E in the retinal pigment epithelium. Arch BiochemBiophys, 483: 175181 Bisti S, Maccarone R, Falsini B. 2014. Saffron and retina: neuroprotection and pharmaco-kinetics. Visual Neurosci, 1-7. doi: 10.1017 / SO952523814000108. Chàbera P, Fuciman M, Hribek P, Polivka T. 2009. Effect of carotenoid structure on excited-state dynamics of carbonyl carotenoids.PhysChemChemPhys, 11: 87958803. Elliott JG, NS Williams. 2012. Nutrients in the battle against age-related eye diseases. American Optometric Association. doi: 10.1016 / j.optm.2011.11.006 Falsini B, Piccardi M, Minnella A, Savastano C, Capoluongo E, Fadda A, Balestratti 20 E, Maccarone R, Bisti S. 2010. Influence of saffron supplementation on retinal flicker sensitivity in early age-related macular degeneration. InvestOphthalmol Screw Sci, 51: 6118-6124. Fernàndez-Sanchez L, Lax P, Esquiva G, Martin-Nieto J, Pinilla I, Cuenca N. 2012. Safranal, a saffron constitute, attenuatesretinaldegeneration in P23H rats. PLoS 25 ONE, 7 (8): e43074. Fontaine V, Lafont R, JA Sahel, Veillet S. 2011. Use of compounds and composition for the treatment of age-related macular degeneration (AMD). Application FR 25506 (filed May 14, 2011). Goralczyk R, Buser S, Bausch J, Bee W, Zühlke U, Barker FM. 1997. Occurrence of 30 birefringent retinal inclusions in cynomolgus monkeys after high doses of canthaxanthin. Invest Ophthalmol Screw Sci, 38: 741-752. Hagiwara A, May N, Ichihara T, Sano M, Tamano S, Aoki H, Yasuhara K, Koda T, Nakamura M, Shirai T. 2003. A thirteen-week oral toxicity study of annatto extract 303 5 5 8 9 16 ( norbixin), a natural food color extracted from the seed of annatto (Bixaorellana L.), in Sprague-Dawley rats. Food ChemToxicol, 41: 1157-1164. Laabich A, Vissvesvaran GP, KL Location, Murata K, McGinn TE, Manmoto CC, Sinclair JR, Karliga I, Leung DW, Fawzi A, Kubota R. 2006. Protective effect of crocin against 5 light blue and white light-mediated photoreceptor cell death in cattle and primate retinal primary cell culture. Invest Ophthalmol Screw Sci, 47: 3156-3163. Levy LW, Regalado E, Navarette S, Watkins RH. 1997. Bixin and norbixin in human plasma: Determination and study of the absorption of a single dose of annatto food color. Analyst, 122: 977-980.
    [0022] Liu X, Osawa T. 2007. Cisastaxanthin and especially 9-cisastaxanthin exhibits a higher antioxidant activity in vitro compared to all-trans isomer. BiochemBiophys Res Comm, 357: 187-193. Maccarone R, Di Marco S., Bisli S. 2008. Saffron supplementation maintains morphology and function after exposure to damaging light in mammalian retina.
    [0023] 15 Invest Ophthalmol Screw Sci, 49: 1254-1261. Maeda T, Maeda A, Golczak M, Palczewski K. 2008. Retinopathy in mice induced by disrupted all-trans-retinal clearance. J Biol Chem, 283: 26684-26693. Maeda T, Maeda A, Matosky M, Okano K, Roos S, Tang J, Palczewski K. 2009. Evaluation of potential therapies for a mouse model of human aging-related macular degeneration caused by delayed all-trans-retinal clearance.tnvest Ophthalmol Screw Sci, 50: 4917-1925. Melendez-Martinez AJ, CM Stinco, Liu C, Wang XD. 2013. A simple HPLC method for the comprehensive analysis of cis / trans (Z / E) geometrical isomers of carotenoids for nutritional studies. Food Chem, 138: 1341-1350.
    [0024] 25 Montenegro MA, De O Rios A, AZ Mercadante, Nazareno MA, Borsarelli CD. 2004. Model studies on the photosensitized isomerization of bixin. J Agric Food Chem, 52: 367-373. Parisi V, Tedeschi M, Gallinaro G, Varano M, Saviano S, Piermarocchi S. 2008. Carotenoids and antioxidants in age-related maculopathy Italian study: multifocal 30 electroretinogram changes after 1 year. Ophthalmology, 115 (2): 324-333. Phan-Thi H, Waché Y. Isomerization and increase in the antioxidant properties of lycopene from Momordicacochinchinensis (gac) by moderate heat treatment with UV-Vis spectra a marker. Food Chem, 156: 58-63.
    [0025] 3035589 17 Pinazo-duran MD, Gomez-Ulla F, Arias L, Araiz J, Casaroli-Marano R, GallegoPinazo R, Garcia-Medina JJ, Lopez-Gavevez MA, Manzanaq L, Salas A, Zapara M, Diaz-Llopis M, Garcia-Layana A. 2014. Do nutritional supplements have a negative impact on macular degeneration prevention J Ophthalmology, article ID 901686.
    [0026] 5 Pintea A, Rugina DO, Pop R, Bunea A, Socaciu C. 2011. Xanthophylls protect against induced oxidation in cultured human retinal pigment epithelial cells. J Food Compos Anal, 24 (6): 830-836. ADO Rios, Borsarelli CD, Mercadante AZ. 2005. Thermal degradation kinetics of bixin in an aqueous model system. J Agric Food Chem, 53: 2307-2311.
    [0027] Sparrow JR, Cai B. 2001. Blue light-induced apoptosis of A2E-containing RPE: involvement of caspase-3 and protection by Bcl-2. Invest Ophthalmol Screw Sci, 42: 1356-1362. Subczynski WK, Wisniewska A, Widomska J. 2010. Location des macular pigments dans la plus vulnerable regions de photoreceptor outer-segment membranes.Arch BiochemBiophys, 504: 61-66. Tsuruma K, Shimazaki H, Nakashima K, Yamauchi M, Sugitani S, Shimazawa M, Linuma M, Hara H. 2012. Annatto prevents retinal degeneration induced by endoplasmic reticulum stress in vitro and in vivo.MolNutr Food Res, 56: 713-724 . Veillet S, Lafont R, Dioh W. 2009. Cosmetic composition for protection of the sun containing urucumextract.Priority Application FR2009-54354 A (June 25, 2009), Application No. FR 2009-54354, WO 2010-FR51323. Verma RS, Middha D. 2010.Analysis of saffron (Crocus sativusL.) Stigma components by LC-MS-MS. Chromatographia, 71: 117-123. Widomska J, Subczynski WK. 2014. Why do you want to protect the retina J ClinExpOphthalmol, 5 (1): 326, doi: 10: 4172 / 2155-9570.1000326. Wu L, Ueda K, Nagasaki T, Sparrow JT. 2014. Light damage in Abca4 and Rpe65rd / 2 mice. Invest Ophthalmol Screw Sci, 55: 1910-1918. Yamauchi M, Tsuruma K, May S, Nakanishi T, Umigai N, Shimazawa M, Hara H. 2011. Crocetin prevents retinal degeneration caused by oxidative stress and endoplasmic reticulum stress via inhibition of caspase activity. Mol Cell Pharmacol, 650: 110-119. JP Young, Zhou J, Nakanishi K, Sparrow JN. Anthocyanins protect against A2E photooxidation and membrane permeabilization in retinal pigment epithelial cells. PhotochemPhotobiol, 81: 529-536.
    权利要求:
    Claims (10)
    [0001]
    REVENDICATIONS1. A composition containing norbixin obtained by purification from a seed extract of Bixa orellana for use in photoprotecting retinal pigment epithelium (RPE) cells in the mammal.
    [0002]
    2. Composition according to claim 1, comprising more than 90% by weight of Norbixin.
    [0003]
    3. Composition according to one of claims 1 or 2, comprising more than 90% by weight of Norbixin in its 9'-cis form of formula (I) CH 3 CH 3 CH 3 CH 3 (I)
    [0004]
    4. Composition according to one of claims 1 to 3, comprising at least one element selected from zinc, vitamin C and vitamin E.
    [0005]
    5. Composition according to one of claims 1 to 4, in the form of a food supplement or a drug.
    [0006]
    6. Composition according to one of claims 1 to 5, comprising a carrier acceptable to be ingested, injected into the eye, injected systemically or injected into the blood.
    [0007]
    7. Composition according to one of claims 1 to 6, for its use for the protection of the cells of the EPR by administration to the mammal, per day, in an amount of between 0.48 mg / kg body weight and 48 mg / kg. kg of body weight, preferably between 0.6 mg / kg of body weight and 20 mg / kg of body weight. 3035589 19
    [0008]
    8. Composition according to one of claims 1 to 7, for its application in the prevention of damage to the retina caused by exposure to blue radiation corresponding to the blue band of the spectrum of visible light, wavelength between 435 nm and 490 nm.
    [0009]
    9. Composition according to one of claims 1 to 8, for its application in the treatment of age-related oracular degeneration (AMD) in mammals. 10
    [0010]
    10. Composition according to one of claims 1 to 8, for its application in the treatment of Stargardt's disease and / or retinitis pigmentosa in mammals.
    类似技术:
    公开号 | 公开日 | 专利标题
    Reinisalo et al.2015|Polyphenol stilbenes: molecular mechanisms of defence against oxidative stress and aging-related diseases
    Kijlstra et al.2012|Lutein: more than just a filter for blue light
    KR101660195B1|2016-09-26|Krill oil and reacted astaxanthin composition and associated method
    EP2717891B1|2016-09-14|BIXA ORELLANA COMPOSITION FOR TREATING MACULAR DEGENERATION&amp; xA;
    EP3288551B1|2019-07-31|Composition containing norbixin for the protection of retinal pigment epithelial cells
    US9295698B2|2016-03-29|Krill oil and carotenoid composition, associated method and delivery system
    US9351982B2|2016-05-31|Krill oil and reacted astaxanthin composition and associated method
    EP3273947B1|2019-06-12|Composition for ocular health
    Fontaine et al.2016|Norbixin protects retinal pigmented epithelium cells and photoreceptors against A2E-mediated phototoxicity in vitro and in vivo
    EP2726087A1|2014-05-07|Compositions containing broccoli seeds for treating or preventing prostate cancer
    FR3101772A1|2021-04-16|composition comprising carotenoids and their use for the protection of neurons against neurodegeneration
    CA3005527A1|2017-05-26|Mirabegron for the treatment of retinal diseases
    同族专利:
    公开号 | 公开日
    US20180289651A1|2018-10-11|
    RU2715889C2|2020-03-04|
    RU2017141462A3|2019-09-12|
    IL255276D0|2017-12-31|
    WO2016174360A1|2016-11-03|
    EP3288551A1|2018-03-07|
    JP2018518460A|2018-07-12|
    CN107708685A|2018-02-16|
    KR20180011777A|2018-02-02|
    FR3035589B1|2019-12-13|
    PL3288551T3|2020-04-30|
    RU2017141462A|2019-05-31|
    CN107708685B|2021-09-21|
    BR112017023264A2|2018-08-07|
    US10314804B2|2019-06-11|
    CA2984405A1|2016-11-03|
    AU2016256637A1|2017-12-14|
    WO2016174360A9|2017-11-16|
    EP3288551B1|2019-07-31|
    MX2017013918A|2018-04-24|
    ES2752061T3|2020-04-02|
    JP6660401B2|2020-03-11|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    WO2001085183A2|2000-05-08|2001-11-15|N.V. Nutricia|Preparation for the prevention and treatment of ocular disorders|
    WO2005110375A1|2004-05-08|2005-11-24|Paul Edward L Jr|Nutritional supplement for treatment of ocular diseases|
    JP2010285364A|2009-06-10|2010-12-24|Riken Vitamin Co Ltd|Singlet oxygen scavenger|
    FR2947173A1|2009-06-25|2010-12-31|Inst Biophytis|FOOD COMPOSITION INTENDED FOR SOLAR PROTECTION|
    FR2975008A1|2011-05-13|2012-11-16|Inst Biophytis|USE OF COMPOUNDS AND COMPOSITION FOR THE TREATMENT OF AGE-RELATED MACULAR DEGENERATION |
    FR1154172A|1955-07-05|1958-04-03|Rollei Werke Franke Heidecke|protective case for cameras|KR20200134935A|2019-05-24|2020-12-02|비지엔케어|The pharmaceutical composition for improvement of human retinal pigment epithelial cell comprising curcumin as an active ingredient|
    FR3105790B1|2019-12-26|2022-01-14|Biophytis|Chemical compounds targeting the eye and their use in the treatment of ocular diseases|
    KR20210008884A|2021-01-08|2021-01-25|비지엔케어|Composition for improving human retinal pigment epithelial cell proliferation containing curcumin as an active ingredient and functional food containing the same|
    法律状态:
    2016-05-02| PLFP| Fee payment|Year of fee payment: 2 |
    2016-11-04| PLSC| Search report ready|Effective date: 20161104 |
    2017-05-02| PLFP| Fee payment|Year of fee payment: 3 |
    2017-07-14| CJ| Change in legal form|Effective date: 20170608 |
    2017-07-14| TQ| Partial transmission of property|Owner name: BIOPHYTIS, FR Effective date: 20170608 Owner name: UNIVERSITE PARIS 6 PIERRE ET MARIE CURIE, FR Effective date: 20170608 |
    2017-07-14| CD| Change of name or company name|Owner name: BIOPHYTIS, FR Effective date: 20170608 Owner name: UNIVERSITE PARIS 6 PIERRE ET MARIE CURIE, FR Effective date: 20170608 |
    2018-05-02| PLFP| Fee payment|Year of fee payment: 4 |
    2019-04-29| PLFP| Fee payment|Year of fee payment: 5 |
    2019-12-27| TQ| Partial transmission of property|Owner name: SORBONNE UNIVERSITE, FR Effective date: 20191120 Owner name: BIOPHYTIS, FR Effective date: 20191120 |
    2020-04-30| PLFP| Fee payment|Year of fee payment: 6 |
    2021-04-30| PLFP| Fee payment|Year of fee payment: 7 |
    优先权:
    申请号 | 申请日 | 专利标题
    FR1553957|2015-04-30|
    FR1553957A|FR3035589B1|2015-04-30|2015-04-30|COMPOSITION FOR THE PROTECTION OF CELLS OF THE RETINAL PIGMENTARY EPITHELIUM|FR1553957A| FR3035589B1|2015-04-30|2015-04-30|COMPOSITION FOR THE PROTECTION OF CELLS OF THE RETINAL PIGMENTARY EPITHELIUM|
    US15/570,720| US10314804B2|2015-04-30|2016-04-28|Composition containing norbixin for protecting cells of the retinal pigment epithelium|
    RU2017141462A| RU2715889C2|2015-04-30|2016-04-28|Composition containing norbixin for protecting cells of retinal pigment epithelium|
    EP16722319.7A| EP3288551B1|2015-04-30|2016-04-28|Composition containing norbixin for the protection of retinal pigment epithelial cells|
    JP2017556593A| JP6660401B2|2015-04-30|2016-04-28|Norbixin-containing composition for protecting cells of retinal pigment epithelium|
    MX2017013918A| MX2017013918A|2015-04-30|2016-04-28|Composition containing norbixin for protecting cells of the retinal pigment epithelium.|
    PL16722319T| PL3288551T3|2015-04-30|2016-04-28|Composition containing norbixin for the protection of retinal pigment epithelial cells|
    CN201680037407.9A| CN107708685B|2015-04-30|2016-04-28|Composition containing norbixin for protecting retinal pigment epithelial cells|
    AU2016256637A| AU2016256637A1|2015-04-30|2016-04-28|Composition containing norbixin for protecting cells of the retinal pigment epithelium|
    ES16722319T| ES2752061T3|2015-04-30|2016-04-28|Composition containing norbixin for the protection of retinal pigment epithelial cells|
    PCT/FR2016/051001| WO2016174360A1|2015-04-30|2016-04-28|Composition containing norbixin for protecting cells of the retinal pigment epithelium|
    CA2984405A| CA2984405A1|2015-04-30|2016-04-28|Composition containing norbixin for protecting cells of the retinal pigment epithelium|
    BR112017023264-2A| BR112017023264A2|2015-04-30|2016-04-28|composition|
    KR1020177034255A| KR20180011777A|2015-04-30|2016-04-28|Norvixin-containing composition for protecting retinal pigment epithelial cells|
    IL255276A| IL255276D0|2015-04-30|2017-10-26|Composition containing norbixin for protecting cells of the retinal pigment epithelium|
    [返回顶部]