BIXA ORELLANA COMPOSITION FOR THE TREATMENT MACULAR DEGENERATION

Abstract

A method for photoprotection of the retinal pigmentary epithelium in a mammal includes administering an effective amount of a composition containing an extract of seeds of Bixa orellana. A process for preparing the extract and formulations of the composition are also provided.

Claims

1. A method for photoprotection of the retinal pigmentary epithelium in a mammal, comprising administering an effective amount of a composition comprising an extract of seeds of Bixa orellana to a subject in need thereof, wherein the extract is prepared according to the following steps: a) extracting the seeds of Bixa orellana in an alcohol solvent to remove a waxy film from the surface of the seeds, and obtaining a suspension in the alcohol; b) removing the seeds from the suspension; c) reducing the suspension and decanting to form a heavy suspension; and d) eliminating a lipid-rich supernatant from the heavy suspension; and e) obtaining a composition that is rich in carotenoids and bixin.

2. The method according to claim 1, wherein the composition is used in a formulation for its application in the treatment of macular degeneration associated with age (AMD) in a mammal.

3. The method according to claim 1, wherein the composition is used in a formulation for its application in the treatment of Stargardt's disease and/or retinitis pigmentosa.

4. The method according to claim 1, wherein the composition is used in a formulation for preventing retinal damage caused by exposure to blue light of wavelength between 435 nm and 490 nm.

5. The method according to claim 1, wherein the composition is comprised in an acceptable carrier to be ingested or injected into the eye or injected into the blood.

6. The method according to claim 1 wherein the extract is comprised in a medicament or food supplement or food.

7. The method according to claim 1, wherein the composition further comprises a gallic acid derivative and/or a compound of the family of anthocyanidins.

8. The method according to claim 7, wherein the gallic acid derivative is ellagic acid.

9. The method according to claim 7, wherein the compound of the family of anthocyanidins is cyanidin.

10. A method for photoprotection of the retinal pigmentary epithelium in a mammal, comprising administering an effective amount of a composition comprising a compound of gallic acid and/or a compound of the family of anthocyanidins.

11. The method according to claim 10, for its application in the treatment of age-related macular degeneration (AMD), or in treating Stargardt's disease and/or retinitis pigmentosa, or to prevent the retinal damage caused by exposure to blue light of wavelength between 435 nm and 490 nm.

12. The method according to claim 10, wherein the gallic acid derivative is ellagic acid.

13. The method according to claim 10, wherein the compound of the family of anthocyanidins is cyanidin.

14. The method according to claim 10, wherein the composition is comprised in a medicament or food supplement or food.

15. The method according to claim 1, wherein the extract is subjected to saponification to turn all or part of the bixin to norbixine.

16. The method according to claim 1, wherein the preparation of the extract further comprises step f) spray drying the composition obtained in step e).

17. The method according to claim 1, wherein the composition comprises 16% by weight of bixin.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] FIG. 1 illustrates the effect of an extract urucum of bixin and norbixin on the protection of the EPR tested for phototoxicity.

[0028] FIG. 2 illustrates the effect of chlorogenic acid, rutin and of ellagic acid on the protection of the EPR tested for phototoxicity.

[0029] FIG. 3 illustrates the effect of cyanidin, cyanidin-3-glucoside or delphinidin-3-sambubioside and 20-hydroxyecdysone on the protection of the EPR tested for phototoxicity.

EMBODIMENTS OF THE INVENTION

[0030] I. Preparing an Extract of Bixa orellana (Extract A)

[0031] Extract A is made by stirring the seeds urucum in absolute ethanol (3 L per kg of seeds) for 16 hours. The agitation in alcohol has the effect of detaching the waxy film on the surface lying seeds.

[0032] A suspension is obtained. It is sieved to remove urucum seeds. This suspension is then reduced to ⅛th then decanted. A heavy suspension is formed.

[0033] The lipid-rich supernatant is eliminated. Maltodextrin is added to the solid deposit and the mixture is spray dried.

[0034] The extract contains 16% by weight of bixin. In the following examples, the concentration of the extract is expressed in bixin equivalents.

[0035] Extract A is also rich in carotenoids. It also contains other terpenic compounds such as geranylgeraniol and tocotrienols (90% δ and 10% β) and several flavonoids.

[0036] The extract has the following features for 100 g (Table 1):

TABLE-US-00001 TABLE 1 Energetic value 319.04 Kcal Carbohydrates 38.10 g (by difference) Bixin 16 g Protein 7.7 g Fat (by hydrolysis) 1.6 g Toral fibers 27.9 g Sodium 8.2 mg Humidity   6% Minerals 4.3%

[0037] According to one embodiment of the invention, the extract A can be subjected to saponification, so as to turn all or part of bixin to norbixin.

II. Activity Assays

[0038] The inventors have tested 15 natural substances and extract A on a RPE cellular model of phototoxicity described below (Table 2).

TABLE-US-00002 TABLE 2 Type Compound name Source (example) Positive Lutein Spinach controls Zeaxanthin Maize Resveratrol Grape Carotenoids Bixin Urucum Norbixin Urucum Extract A Urucum Crocetin Saffron Phenolic acids Chlorogenic acid Mate Flavone Orientin Açaï Flavonol Rutin Buckwheat Flavanone Naringenin Lemon Benzopyrane Ellagic acid Pomegranate Anthocyanins Cyanidin 3-glucoside Açaï Delphinidin 3-sambubioside Hibiscus Anthocyanidins Cyanidin Açaï* Steroids 20-Hydroxyecdysone Quinoa *The cyanidin is prepared after acid hydrolysis of its glycosylated forms

[0039] To test the photoprotective effect of the test substances, the inventors used a cellular model of induced phototoxicity by treatment with A2E followed by illumination with blue radiation. This model was produced from primary cultures of adult porcine RPE cells. Cell survival was determined by the ratio between the number of living cells and the total number of cells (alive+dead, respectively quantified using specific staining). Image acquisition was performed using a fluorescence microscope controlled by Metamorph software and quantifications were made by processing images acquired by a program dedicated quantification. The experiments were performed in 96-well microplates in quadruplicate and the experiment was repeated at least four times. Cells were treated for 48 hours with these compounds, the last 24 hours in the presence of A2E before induction of phototoxicity. Three concentrations (0.1, 1 and 10 μM) were tested for each compound. Some compounds were further tested at 20 μM to achieve a range of concentrations.

III. Results

[0040] The results, presented as averages and standard deviations are expressed as percentage of survival compared to the control without A2E.

[0041] The experiments did not show a protective effect of 15 compounds or extract A at concentrations of 0.1 and 1 μM (data not shown for lutein, zeaxanthin, resveratrol, crocetin, naringenin and orientin, and results shown for urucum extract A, cyanidin and ellagic acid in FIGS. 1-3).

[0042] In a series of tests (n=5), extract A allowed a significant protection, with cell survival for 20 μM of the order of 93% of the control without A2E, to be compared with that of control+A2E, which is only 45% (FIG. 1).

[0043] Among the 15 compounds tested at 10 μM, two provide a cellular protection against phototoxicity.

[0044] Notably, 10 μM cyanidin provides a protection in the order of 87% survival compared to the control without A2E. At 20 μM, the same compound provides almost complete protection (FIG. 2).

[0045] Ellagic acid also provides protection in the order of 68% compared to the control without A2E. At 20 μM, this compound provides protection similar to that measured at 10 μM, but with a better reproducibility (FIG. 3).

[0046] Photoprotective effects expected for lutein, zeaxanthin and resveratrol (“positive controls”) were not observed with these experimental conditions.

REFERENCES

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