CHEMICAL COMPOUNDS TARGETING THE EYE AND USE THEREOF IN THE TREATMENT OF EYE DISEASES

Abstract

Disclosed is chemical compounds C that are derivatives of norbixin and have tropism for the eye, and are intended to be used in the treatment of eye diseases in mammals, in particular in the context of altering the retinal pigment epithelium and more particularly in the context of age-related macular degeneration (AMD) and Stargardt's disease.

Claims

1. Chemical compound having the following general formula (I): ##STR00034## where COR is a secondary or tertiary amide, such that —R is chosen from: -M; —NH—(CH.sub.2).sub.n-M and —NH—(CH.sub.2).sub.n—C(CH.sub.3)(CH.sub.3)-M, -M being chosen from a) ##STR00035## wherein, R.sup.1 is chosen from an oxygen atom, a sulfur atom, a >CH.sub.2, >CH—O—(CH.sub.2).sub.n—CH.sub.3, >CH—(CH.sub.2).sub.n—O—(CH.sub.2).sub.n—CH.sub.3, >CH—(CH.sub.2).sub.n—OH, >CH—COOH, >C(OH)phenyl or >NH group; R.sup.3 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; R.sup.4 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; n is an integer comprised between 0 and 6; —NH—(CH.sub.2).sub.n—W, W being a hydrogen atom, or a —OH group, a —O—(CH.sub.2).sub.n—CH.sub.3 group, or a group chosen from i) ##STR00036## wherein, R.sup.1 is chosen from an oxygen atom, a sulfur atom, a >CH.sub.2, >CH—O—(CH.sub.2).sub.n—CH.sub.3, >CH—(CH.sub.2).sub.n—OH, >CH—COOH, >C(OH)phenyl or >NH group; R.sup.3 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; R.sup.4 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; R.sup.5 is chosen from a —CH.sub.3, —OH, —O—(CH.sub.2).sub.n—CH.sub.3, —(CH.sub.2).sub.n—OH or —COOH group; n is an integer comprised between 0 and 6; as well as the pharmaceutically acceptable salts of said chemical compound.

2. Chemical compound having the general formula (I) according to claim 1, chosen from the following chemical compounds: 1-[2-methoxyethanamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[1,4-oxazinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[piperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[2-hydoxyethanamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1[1,4-oxazepanamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-thiomorpholinamido-(2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-pyrrolidinamido-(2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[2-morpholinopropanamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[(S)-3-hydroxypyrrolidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[2-morpholinoethanamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[(R)-3-hydroxypyrrolidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-hydroxypiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[2-methyl-2-(4-morpholinyl)propylamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-hydroxymethylpiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[(Z)-2,6-dimethylmorpholinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-hydroxyphenylamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-benzyl-4-hydroxypiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-carboxypiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[(E)-4-hydroxycyclohexamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[3-methoxypiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-methoxypiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[2-(2-furyl)ethanamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-n-propoxypiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate; 1-[4-ethylmethoxypiperidinamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate.

3. Composition comprising at least one said chemical compound according to claim 1.

4. The composition according to claim 3 comprising at least one excipient.

5. The composition according to claim 3, comprising a support in a form adapted to be ingested, injected into the eye or injected into the blood.

6. A drug comprising the chemical compound having the general formula (I) according to claim 1.

7. A method for the photoprotection of the cells of the retinal pigment epithelium in mammals, comprising administering to said cells an effective amount of the chemical compound having the general formula (I) of claim 1.

8. A method for the treatment and/or the prevention of damage to the retina of mammals caused by exposure to blue light corresponding to the blue band of the visible light spectrum, of a wavelength comprised between 435 nm and 490 nm, comprising administering an effective amount of the chemical compound having the general formula (I) of claim 1 to a patient in need thereof.

9. A method for the treatment and/or the prevention of damage to the retina of mammals caused by retinal degenerations, comprising administering an effective amount of the chemical compound having the general formula (I) of claim 1 to a patient in need thereof.

10. A method for treatment and/or the prevention of eye diseases in mammals, comprising administering an effective amount of the chemical compound having the general formula (I) of claim 1 to a patient in need thereof.

11. A method for treatment and/or the prevention of retinopathies in mammals, comprising administering an effective amount of the chemical compound having the general formula (I) of claim 1 to a patient in need thereof.

12. A method for treatment and/or the prevention of age-related macular degeneration (AMD) in mammals, comprising administering an effective amount of the chemical compound having the general formula (I) of claim 1 to a patient in need thereof.

13. A method for treatment and/or the prevention of Stargardt's disease and Retinitis pigmentosa in mammals, comprising administering an effective amount of the chemical compound having the general formula (I) of claim 1 to a patient in need thereof.

14. Composition comprising at least one said chemical compound according to claim 2.

15. The composition according to claim 4, comprising a support in a form adapted to be ingested, injected into the eye or injected into the blood.

16. A drug comprising the chemical compound having the general formula (I) according to claim 2.

17. A drug comprising composition of claim 3.

18. A method for the photoprotection of the cells of the retinal pigment epithelium in mammals, comprising administering to said cells an effective amount of the chemical compound having the general formula (I) of claim 2.

19. A method for the photoprotection of the cells of the retinal pigment epithelium in mammals, comprising administering to said cells an effective amount of the composition of claim 3.

20. A method for the treatment and/or the prevention of damage to the retina of mammals caused by exposure to blue light corresponding to the blue band of the visible light spectrum, of a wavelength comprised between 435 nm and 490 nm, comprising administering an effective amount of the chemical compound having the general formula (I) of claim 2 to a patient in need thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0064] The invention shall be better understood when reading the following description, given as an example and in no way limiting, and given in reference to the figures which show:

[0065] FIG. 1 FIG. 1 shows pharmacokinetic profiles of compounds object of the present invention in the eye. These are graphs showing the ocular concentrations as a function of time after administration of the Chemical compounds (C) object of the present invention and of norbixin (BIO201). All the compounds were administered by mouth (p.o.) at 50 mg/kg formulated in a medium D-α-Tocopheryl polyethylene glycol 1000 succinate (VitE-TPGS) 20% in a sodium bicarbonate buffer (0.1M). The pharmacokinetic profiles of compounds having a low ocular exposure are shown in the graph A of FIG. 1, those having an average ocular exposure are shown in the graph B of FIG. 1, and those having a high ocular exposure are shown in the graph C of FIG. 1.

[0066] FIG. 2 FIG. 2 shows pharmacokinetic profiles of compounds object of the present invention in the plasma. These are graphs showing the plasma concentrations as a function of time after administration of the Compounds (C) and norbixin (BIO201). All the compounds were administered by mouth (p.o.) at 50 mg/kg formulated in a medium VitE-TPGS 20% in a sodium bicarbonate buffer (0.1M). The plasma exposure profiles of compounds having a low ocular exposure are shown in the graph A of FIG. 2, those of the compounds having an average ocular exposure are shown in the graph B of FIG. 2, and finally those of the compounds having a high ocular exposure are shown in the graph C of FIG. 2.

[0067] FIG. 3 FIG. 3 shows a chronogram of the photoprotection experiments of the cells of the retinal pigment epithelium (RPE) by compounds (C) object of the present invention in vitro. This is the chronology of the steps implemented so as to test the photo-protective activity of the various chemical compounds (C) compared to that of native norbixin on the cells of the RPE put into the presence of A2E and subjected to an illumination.

[0068] FIG. 4 FIG. 4 shows the targeting of the eye and the percentage of photoprotection in vitro of the compounds object of the present invention. These are histograms showing in a combined manner the areas under the curve of the ocular concentrations (ocular exposure) of norbixin (BIO201) and of various compounds C object of the present invention, and the percentages of photoprotection obtained with norbixin and the compounds object of the present invention at 5 μM, 10 μM and 20 μM. The compounds having a low ocular exposure are shown in histogram A, those having an average ocular exposure are shown in histogram B, and those having a high ocular exposure are shown in histogram C.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0069] The chemical compounds object of the present invention do not exist in the chemical databases of carotenoids and di-apo-carotenoids. They are synthesized according to processes that can be industrialized, i.e. with a minimum of synthesis steps and an optimum yield. They have a better pharmacokinetic profile and a better tropism for the eye than those of norbixin. Some of these compounds moreover have a photo-protective activity of the RPE superior than that of norbixin.

Description of Syntheses and General Diagrams

[0070] The chemical compounds having general formula (I):

##STR00004##

can be prepared by application or adaptation of any method known per se to a person skilled in the art and/or within the scope of the latter, which have been able to be described, or by application or adaptation of methods described in the following procedures.

[0071] In the description that follows the various groups refer to the definitions given hereinabove, i.e.:

COR is a secondary or tertiary amide, such that —R is chosen from: [0072] -M; —NH—(CH.sub.2).sub.n-M and —NH—(CH.sub.2).sub.n—C(CH.sub.3)(CH.sub.3)-M, -M being chosen from [0073] a)

##STR00005## [0074] wherein, [0075] R.sup.1 is chosen from an oxygen atom, a sulfur atom, a >CH.sub.2, >CH—O—(CH.sub.2).sub.n—CH.sub.3, >CH—(CH.sub.2).sub.n—O—(CH.sub.2).sub.n—CH.sub.3, >CH—(CH.sub.2).sub.n—OH, >CH—COOH, >C(OH)phenyl or >NH group; [0076] R.sup.3 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; [0077] R.sup.4 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; [0078] n is an integer comprised between 0 and 6; [0079] —NH—(CH.sub.2).sub.n—W, W being a hydrogen atom, or a —OH group, a —O—(CH.sub.2).sub.n—CH.sub.3 group, or a group chosen from [0080] i)

##STR00006## [0081] wherein, [0082] R.sup.1 is chosen from an oxygen atom, a sulfur atom, a >CH.sub.2, >CH—O—(CH.sub.2).sub.n—CH.sub.3, >CH—(CH.sub.2).sub.n—OH, >CH—COOH, >C(OH)phenyl or >NH group; [0083] R.sup.3 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; [0084] R.sup.4 is chosen from a hydrogen atom, a C.sub.1-C.sub.6 alkyl, —OH or C.sub.1-C.sub.6 —O-alkyl group; [0085] R.sup.5 is chosen from a —CH.sub.3, —OH, —O—(CH.sub.2).sub.n—CH.sub.3, —(CH.sub.2).sub.n—OH or —COOH group; [0086] n is an integer comprised between 0 and 6.

[0087] In the framework of the present invention the term “C.sub.1-C.sub.6 alkyl group” means any alkyl group with 1 to 6 carbon atoms, linear or branched, in particular, the methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl, n-pentyl, n-hexyl groups. Advantageously it is a methyl, ethyl, iso-propyl or t-butyl group, in particular a methyl or ethyl group, more particularly a methyl group.

[0088] Diagram A for the synthesis of the compound 1-[2-(2-furyl)ethanamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate (called compound C21):

[0089] Formation of Amide (First Method)

##STR00007##

[0090] Hydrolysis of Methyl Ester

##STR00008##

[0091] Diagram B for the synthesis of the compound 1-[(E)-4-hydroxycyclohexamido](2E,4E,6E,8E,10E,12E,14E,16Z,18E)-4,8,13,17-tetramethylicosa-2,4,6,8,10,12,14,16,18-nonaenedioate (called compound C19:

[0092] Formation of Amide (Second Method)

##STR00009##

DMF=dimethylformamide
TEA=triethylamine
DIA=diisopropylamine
THF=tetrahydrofuran
CDI=carbonyldiimidazole
HBTU=(1H-benzotriazol-1-yloxy)(dimethylamino)-N,N-dimethylmethaniminium hexafluorophosphate

EXAMPLES

[0093] Equipment and Methods

[0094] The nuclear magnetic resonance (NMR) spectra of the proton (.sup.1H) are carried out on a Bruker Avance DPX500 device (500, 0.7 MHz). The chemical shifts (δ) are measured in parts per million (ppm). The spectra are calibrated on the chemical shift of the deuterated solvent used. The coupling constants (J) are expressed in Hertz (Hz) and the multiplicity is represented in the following way, singlet (s), doublet (d), doublet of doublet (dd), triplet (t), triplet of doublet (td), quadruplet (q), multiplet (m). The mass spectra (MS) are carried out with an Agilent Technologies MSD spectrometer, G1946A type, the samples are ionized by an “Atmospheric pressure chemical ionization” (APCI) source.

[0095] By way of illustrating examples of the invention, the molecules shown in table 1 were synthesized.

TABLE-US-00001 TABLE 1 N.sup.o Chemical structure Chemical name C1 [00010] 1-[2- methoxyethanamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C2 [00011] 1-[1,4-oxazinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C3 [00012] 1-[piperidinamido] (2E,4E,6E,8E,10E, 12E,14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C4 [00013] 1-[2-hydroxyethanamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C5 [00014] 1[1,4-oxazepanamido] (2E,4E,6E,8E,10E, 12E,14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C6 [00015] 1-thiomorpholinamido- (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C7 [00016] 1-pyrrolidinamido- (2E,4E,6E,8E,10E, 12E,14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C8 [00017] 1-[2- morpholinopropanamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C9 [00018] 1-[(S)-3- hydroxypyrrolidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C10 [00019] 1-[2- morpholinoethanamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C11 [00020] 1-[(R)-3- hydroxypyrrolidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C12 [00021] 1-[4- hydroxypiperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C13 [00022] 1-[2-methyl-2- (4-morpholinyl) propylamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C14 [00023] 1-[4- hydroxymethyl- piperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C15 [00024] 1-(Z)-3,5- dimethylmorpholinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C16 [00025] 1-[4- hydroxyphenyl- ethanamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C17 [00026] 1-[4-benzyl-4- hydroxypiperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C18 [00027] 1-[4- carboxypiperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C19 [00028] 1-[(E)-4- hydroxycyclohexamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C20 [00029] 1-[3- methoxypiperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C21 [00030] 1-[2-(2-furyl) ethanamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C22 [00031] 1-[4-n- propoxypiperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C23 [00032] 1-4- ethylmethoxy- piperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)-4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate C24 [00033] 1-4- methoxypiperidinamido] (2E,4E,6E,8E,10E,12E, 14E,16Z,18E)- 4,8,13,17- tetramethylicosa- 2,4,6,8,10,12,14,16,18- nonaenedioate

Example 1 (Diagram A)

[0096] Preparation of the Compound C21

[0097] Step 1 (Typical Coupling Method of an Amine with Carbonyidiimidazole (CDI)): Preparation of the Methyl Ester of C21

[0098] The chemical reactions were carried out under argon atmosphere. The reagents and the products were protected from light with aluminum paper during all the steps of the reaction and during the step of extracting and removing solvents. The products were stored at 4° C. The bixin (1 g; 2.54 mmol) is dissolved in 10 mL of dry dimethylformamide (DMF) and triethylamine (1.06 mL; 7.61 mmol) then CDI (0.823 g; 5.07 mmol) are added. After two hours, 2-aminoethylfuran (845 mg; 7.61 mmol) was added and the reaction mixture was stirred for 18 h at 20° C. Hydrochloric acid (HCl, 1 N; 20 mL) is added and the resulting suspension is centrifuged. Following the removal of the supernatant, the pellet containing the methyl ester was resuspended twice in the presence of water. Following another centrifugation, the pellet (1.24 g) was used directly in the step of hydrolysis.

[0099] Step 2 (Typical Procedure of the Hydrolysis): Conversion of the Methyl Ester of C21 into C21

[0100] The chemical reactions were carried out under argon atmosphere and protected from light with aluminum paper. The products are stored at 4° C. The methyl ester of C-21 is dissolved in 15 mL of tetrahydrofuran (THF) and 13 mL of MeOH. Sodium hydroxide (NaOH, 1 N; 15.3 mL) is added and the reaction mixture is stirred for 20 h at 20° C. Hydrochloric acid (HCl; 1 N; 16 mL) was added progressively. The suspension obtained is centrifuged and the supernatant is removed. The pellet is then resuspended and mixed twice with water then centrifuged again and the supernatant is removed. The paste obtained, containing water, is transferred to a pyriform flask in the presence of water and acetonitrile, and lyophilized to give 1.04 g of an orangish to red colored powder.

[0101] Analyses of the Compound C21

[0102] LC-MS: m/z=474.3 (MH.sup.+) UV purity at 460 nm=99%.

[0103] NMR .sup.1H (500 MHz, DMSO-d.sub.6) −δ 8.11 (t, 1H), 7.54 (s, 1H), 6.19 (d, 1H), 3.42-3.38 (m, 2H), 2.78 (t, 2H).

[0104] Preparation of the Compound C19

[0105] Step 1 (Typical Procedure for the Coupling of the Amine by Using HBTU): Preparation of the Methyl Ester of C19 (Method Used for the Preparation of the Methyl Ester of C19 Only, as the Reaction Using CDI is Too Slow for this Compound)

[0106] The chemical reactions were carried out under argon atmosphere. The reagents and the products were protected from light with aluminum paper during all the steps of the reaction and during the step of extracting and removing solvents. All the products were stored at 4° C. The bixin (1.17 g; 2.97 mmol) is dissolved in 20 mL of anhydrous DMF and diisopropylamine (1.23 mL; 7.42 mmol); HBTU ((1H-benzotriazol-1-yloxy)-dimethylamino hexafluorophosphate)-N, N′-dimethylmethaniminium; 1.69 g; 4.45 mmol) is then added. After 1 h, trans-4-aminocyclohexanol (680 mg; 4.45 mmol) was added and the reactive mixture is stirred for 18 h at 20° C. Sixty milliliters of water (60 mL) is added and the precipitate was filtered and washed twice with 60 mL of water to give 1.4 g of violet solid compound which is used as is in the step of hydrolysis.

[0107] Step 2 (Typical Procedure of the Hydrolysis): Conversion of the Methyl Ester of C19 into C19

[0108] The chemical reactions were carried out under argon and protected from light with aluminum paper. The products are stored at 4° C. The 1.4 g of methyl ester of C-21 are dissolved in 10 mL of tetrahydrofuran (THF) and 10 mL of MeOH (dark red solution). Five equivalents of sodium hydroxide (NaOH, 10N; 1.43 mL) are added and the reaction mixture is stirred for 48 h at ambient temperature. In HPLC-MS, a single peak is observed and the absence of starting material. Hydrochloric acid (HCl 12N; 1.18 mL) was added. The precipitate obtained is diluted with 60 mL of water. The solution is centrifuged and the aqueous supernatant is removed. The solid pellet is then taken and mixed with water (30 mL) then centrifuged again and the supernatant is removed. This step is repeated twice until a solid paste of orange color is obtained (which forms a suspension in the presence of water). The solid paste is taken in 50 mL of water to give an orange suspension, frozen and directly lyophilized to give an orange powder (1.02 g).

[0109] Analyses of the Compound C19

[0110] LC-MS: m/z=478.2 (MH.sup.+) UV purity at 460 nm=97.3%.

[0111] NMR .sup.1H (500 MHz, DMSO-d.sub.6) −δ 47.82 (s, 1H), 4.51 (m, 1H), 3.58.-3.53 (m, 1H), 3.40-3.36 (m, 1H), 1.83-1.75 (m, 4H), 1.26-1.15 (m, 4H).

[0112] Cascade Screening and Characterization of the Biological Effects of the Chemical Compounds (C) Derived from Norbixin.

[0113] The development of the screening test was initiated from work in literature and based on the characteristics of the pathology of dry AMD. At the physiopathologic level, this disease is characterized by a progressive loss of induced vision following the degeneration of the photoreceptors and of the cells of the RPE. The cells of the RPE play a crucial role in the survival and correct operation of the photoreceptors by providing them with the necessary nutrients, by participating in the visual cycle and by removing the debris coming from external segments of photoreceptors and that result from this cycle. It is important to screen the drugs in development on their capacity to target preferably the eye while still avoiding an intraocular administration that is traumatizing for patients and having risks of local infections. To do this, a pharmacokinetic study of the chemical compounds (C) derived from norbixin object of the present invention at the plasma and ocular level was conducted so as to select the chemical compounds having an ocular AUC (“area under the curve”) that is improved in relation to norbixin.

[0114] In addition to better distribution in the target-tissue, selecting new compounds must also be based on good photo-protective activity that limits the loss of the cells of the RPE and thus reducing the retinal degeneration observed during the AMD and other degenerative diseases such as Retinitis pigmentosa and Stargardt's disease. At the cellular level, on cultures of cells the RPE coming from pig retina, Fontaine et al. (2016) have shown that a treatment with norbixin (BIO201) protects the cells of the RPE against apoptosis following an illumination in the presence of A2E (80% survival 24 h after exposure). This same screening test according to the percentage of photoprotection was used so as to determine its modulation by and the chemical compounds derived from norbixin (C) object of the present invention in comparison with the photo-protective effect of norbixin and characterize these modulations from a statical standpoint.

[0115] Protocols

[0116] Pharmacokinetic Study Via Oral Administration of the Molecules in Mice

[0117] The pharmacokinetic study of the chemical compounds (C) following the oral administration thereof was conducted using mice C57BL/6 (January, 53940 The Genest Saint Isle, France). The chemical compounds derived from norbixin (C) were administered at a dose of 50 mg/kg of body weight. After administration, the blood was sampled at the tail at t=0.25 h; 0.5 h; 1 h; 3 h; 6 h and 8 h. The blood samples were centrifuged and the plasmas taken. The dosage of the plasma samples made it possible to determine pharmacokinetic parameters, namely C.sub.max, which corresponds to the maximum concentration observed after the administration of the molecule, T.sub.max which is the time required to reach maximum concentration after administration of the molecule and the AUC: the area under the curve which corresponds to the plasma exposure (FIG. 2).

[0118] In parallel, the ocular concentrations of the chemical compounds derived from norbixin (C) were dosed in the following way (FIG. 1): both eyes of each mouse were taken in Precellys tubes and stored at −80° C. until the time of dosing. The eyes are then ground in the Precellys tubes with a bench homogenizer, Fast-prep (Fischer Scientific, Hampton, United States) in a mixture of organic solvents, in a first step with 500 μL of chloroform/methanol (1/1, v/v) then 500 μL of chloroform/dichloromethane (1/1, v/v). The supernatants are recovered at each step and transferred to a 96-well plate of 2 mL.

[0119] For the quantification of the chemical compounds C, a calibration curve is carried out with 8 standards (5 to 5,000 ng/mL) in the same organic solvent mixtures and transferred (100 μL) to the 96-well plate of 2 mL.

[0120] The supernatants and standards are evaporated in a EZ2 (Genevac, Ipswich, United Kingdom), without heating, then taken with 100 μL of DMSO/methanol (20:80, v/v) before being transferred to a 96-well plate of 200 μL.

[0121] The LC-MSMS analysis is carried out with a HPLC 1200 Infinity chain (Agilent Technologies, Santa-Clara, United States), a UV detector and a mass spectrometer QQQ6420 (Agilent Technologies Santa-Clara, United States). The injection volume is 5 μL. The chemical compounds C are eluted on an inverse phase column C18 (2.1*50 mm, particles 3 μm; Ace-C18-Excel, AIT) with a gradient of acetonitrile and water (containing 0.1% formic acid) and a flow rate of 0.3 mL/min. The conditions of the gradient can change according to the chemical compound C analyzed. The UV detector analyzes at 460 nm and the mass spectrometer analyzed in Mode MRM—Positive.

[0122] Photoprotection of the Cells of the RPE

[0123] Tests in vitro of photoprotection by the various chemical compounds C of the Cells of the RPE Illuminated in the Presence of A2E

[0124] The test in vitro described hereinabove and intended to study the photo-protective effect of norbixin was used so as to quantify the photo-protective effects of the various chemical compounds derived from norbixin (C) on the cells of the RPE illuminated with blue light in the presence of A2E (FIG. 3). The photo-protective effect of the molecules was evaluated in a phototoxicity cellular model induced by treatment by A2E followed by an illumination with blue light. The term “blue radiation” means the radiation corresponding to the blue band of the visible light spectrum, i.e. of a wavelength comprised between 435 and 490 nm. This model uses primary cultures of RPE of adult pigs. The cell survival is quantified thanks to a test of cellular viability. At −48 h the compounds to be tested (in solution at 5 mM in the DMSO) are added to obtain the final concentrations of 1 to 20 μM) then at −19 h of the A2E (final concentration 30 μM) and the cells (time 0 h) are illuminated. 24 h after the survival of the cells is measured. The acquisition of images, as well as the processing thereof, are carried out using a fluorescence microscope controlled by the Metamorph software and a dedicated quantification program. The experiments are conducted on a 96-well microplate in quadruplicate and the experiment is reproduced at least four times. The results are expressed in the form of a ratio representing the number of living cells in the wells treated by the molecules to be tested, divided by the number of living cells in the controlled wells (treated by the dilution medium without A2E). This test made it possible hereinabove to reveal the photo-protective activity of norbixin (Fontaine et al. 2016).

[0125] Results

[0126] Pharmacokinetic Study of the Chemical Compounds C in Mice

[0127] Table 2 discloses the pharmacokinetic results of the chemical compounds C following the administration p.o. 50 mg/kg in D-α-Tocopheryl polyethylene glycol 1000 succinate (VitE-TPGS) 20% in a sodium bicarbonate buffer (0.1M).

TABLE-US-00002 TABLE 2 Eye Plasma C.sub.Max T.sub.Max Exposure C.sub.Max T.sub.Max Exposure (ng/eye) (h) (ng .Math. h/eye) (μg/mL) (h) (μg .Math. h/mL) BIO201 8.6 0.5 22.4 34.99 0.25 100.7 C1 10.7 0.5 26.6 18.73 0.5 52.66 C2 41 0.5 186.5 34 0.5 176.2 C3 5.1 0.5 19.1 4.05 0.5 10.37 C4 10.2 0.5 30 57.07 1 208.6 C5 31.4 0.5 188.7 37.71 1 299.9 C6 3.9 0.5 23.1 3.95 0.5 10.81 C7 7 0.5 18.5 6.89 0.5 17.29 C8 56.8 1 472.4 59.82 0.5 173.74 C9 53.1 1 452 37.92 1 342.68 C10 64.3 1 533.2 30.42 1 108.1 C11 47.7 1 238.4 35.91 1 190 C12 50.7 1 538.4 29.31 0.5 186.3 C13 60.7 1 461.1 27.89 0.5 139.7 C14 43.2 1 322.5 19.02 0.5 162.2 C15 22.8 0.5 108.5 17.34 0.5 46.99 C16 7.4 0.5 8.7 10.51 1 13.23 C17 18 2 135.2 8.06 1 32.39 C18 27.7 2 307.8 61.65 0.5 948.84 C19 5.7 0.5 37.5 6.62 0.25 22.8 C20 80.4 2 421.9 35.02 1 160.4 C21 1.6 0.25 0.6 6.61 0.25 9.17 C22 42.1 2 358.5 12.83 0.25 37.25 C23 11 2 54.7 5.01 1 24.1 C24 96 1.5 499.1 48.31 0.5 141.9

[0128] Table 3 corresponds to the percentages of photoprotection of the cells of the RPE by the chemical compounds C in vitro: it shows the percentage of cells of RPE surviving in the presence of N-retinyl-N-retinylidene ethanolamine (A2E) and of the various chemical compounds C derived from norbixin (tested at 5, 10 or 20 μM) or of norbixin in the same concentrations after having been subjected to an illumination.

TABLE-US-00003 TABLE 3 Eye Exposure 50 mg/kg p.o. Photoprotection (%) AUC 5 μM 10 μM 20 μM (ng .Math. h/eye) Mean SE Mean SE Mean SE BIO201 22.4 19.2 6.4 34.1 6.6 63.6 4.6 C1 26.6 55.0 4.8 67.7 2.2 77.8 7.1 C2 186.5 33.9 13.7 22.3 12.9 49.1 11.2 C3 19.1 11.3 11.3 68.1 32.1 45.1 10.3 C4 30.0 16.4 14.0 36.0 17.7 67.2 16.4 C5 188.7 11.3 11.3 25.6 23.3 36.0 11.9 C6 23.1 nd 60.6 30.3 19.1 19.1 C7 18.5 5.2 5.2 52.8 14.8 nd C8 472.4 28.6 8.4 33.4 8.2 75.2 6.7 C9 452.0 22.1 9.4 32.0 12.7 63.7 6.1 C10 533.2 15.5 1.1 27.4 27.4 80.3 11.4 C11 238.4 21.1 7.7 39.3 16.7 63.4 8.2 C12 538.4 7.7 3.1 14.3 7.6 59.6 4.3 C13 461.1 8.0 4.0 0.5 0.3 8.0 6.5 C14 322.5 3.1 3.1 46.8 1.2 79.3 6.0 C15 108.5 5.4 4.0 10.0 5.1 58.5 5.7 C16 8.7 70.2 6.1 81.7 1.6 26.8 5.6 C17 135.2 71.7 5.4 67.2 6.4 44.0 4.9 C18 307.8 27.3 11.7 43.1 12.4 63.8 5.9 C19 37.5 60.6 7.5 60.6 7.5 79.9 4.1 C20 421.9 65.6 5.7 65.6 5.7 79.0 5.4 C21 0.6 56.7 7.0 56.7 7.0 79.6 1.1 C22 358.5 64.5 8.2 72.8 5.7 66.0 7.9 C23 54.7 38.4 10.4 59.2 6.8 73.1 8.3 C24 499.1 47.0 15.0 69.0 13.0 84.0 13.6

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