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A New Drug Candidate for the Treatment of Dry AMD. Fontaine Et Al

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A New Drug Candidate for the Treatment of Dry AMD. Fontaine Et Al #1119 BIO201: a new drug candidate for the treatment of dry AMD V. FONTAINE1, E. BRAZHNIKOVA1, E. MONTEIRO1, L. LESAGE1, C. BALDUCCI2, L. GUIBOUT2, L. FERAILLE3, P.-P. ELENA3, J.-A. SAHEL1, S. VEILLET2, R. LAFONT2 1Sorbonne Universités, UPMC Univ Paris 06, INSERM, CNRS, Institut de la Vision, 17 Rue Moreau, 75012 Paris, France 2Biophytis, Parc BIOCITECH, 102 Avenue Gaston Roussel, 93230 Romainville, France / 3IRIS-Pharma, Les Nertières, Allée Hector Pintus, 06610 La Gaude, France PURPOSE RESULTS Dry AMD is a major cause of visual impairment associated with High protection of RPE cells by bixin and norbixin Norbixin bioavailability is much better than that of bixin aging, and no drug treatment is presently available. In Bixin and norbixin were highly collaboration with the Institut de la Vision, Biophytis develops a 100 **** **** **** protective even at 5 mM inducing drug candidate (BIO201) based on norbixin, a di-apo-carotenoid, **** **** **** 80 **** **** **** **** 80.75 and 72.5 % RPE survival as API. We describe here the effects of norbixin on retinal **** pigmented epithelium (RPE) and retina photoprotection in vitro 60 *** respectively whereas lutein, 40 zeaxanthin and crocetin showed 1000 15000 Blood was collected at various and in vivo. A B 20 no protection at this intervals and plasma levels were 800 12000 concentration. At the different ) (ng/ml) measured by HPLC-MS/MS. cell viability (% of cont - A2E) - of cont(% viability cell MATERIALS AND METHODS 0 — 5 10 20 50 5 10 20 50 5 10 20 50 5 10 20 50 5 10 20 50 600 9000 E E Ingested Bixin (A) bioavailability is 2 concentrations tested norbixin A A2 - bixin norbixin lutein zeaxanthin crocetin 400 6000 nt nt + quite low (ca. 2%) and this compound The photo-protective effect of norbixin, bixin, lutein, zeaxanthin o was always slightly less efficient o ( ) norbixin or C C Norbixin (ng/ml) --- is rapidly converted into norbixin by and crocetin was evaluated on primary cultures of porcine RPE than bixin. 200 3000 cells challenged with 30 µM A2E and illuminated with blue light Lutein, zeaxanthin and crocetin were also able to prevent RPE cell death at the highest Bixin ( esterases. Norbixin bioavailability (B) 0 0 (470 nm, 50 min) (N=16). concentrations. The concentrations of the substances are in µM. The positive control (cont – 0 2 4 6 8 10 0 time (hours)5 10 is much better and may become close time (hours) In vivo experiments measured the photo-protective effect of A2E) represents cells treated with DMSO alone. The negative control (cont + A2E) represents to 100% with a suitable dosage form. norbixin one week after blue light damage in the Abca4-/-Rdh8-/- cells treated with A2E but not with substances. Norbixin undergoes significant conjugation as glucuronide(s) in mouse plasma. transgenic mouse model by intravitreal injections and in a standard rat blue light model of photodamage by repeated -/- -/- intraperitoneal injections. Twenty-eight 7-week-old mice were Norbixin protects the retina of Abca4 Rdh8 mouse against blue light induced phototoxicity injected intra-vitreally in one eye with 120 µM norbixin (in 0.3% 12 DMSO) or with DMSO alone, dark-adapted during 24 hours and RPE norbixin i.e. OS 10 layers norbixin n.i.e. exposed to blue light (4000 lux) for one hour. Ten mice were used ## ## ### ### dmso i.e. ONL 8 as non-illuminated controls. Groups of 6-12 rats were injected ## #### ## # #### ### ### # *** ** ### *** ** dmso n.i.e. ### #### **** #### ****** intraperitoneally with either norbixin (10, 50, 100 mg/kg), α- ###### ######## 6 ** ****** INL ** ######## control ######## #### ****** ### #### ### ******** *** ###### ****** *** **** phenyl-N-tert-butylnitrone (PBN, a potent free-radical trapping photoreceptor **** 4 of agent, 50 mg/kg), or an equivalent volume of saline 30 min prior to GCL light damage and 2, 4 and 6 hours after the beginning of the 2 Number exposure. 18 µm 0 Full field scotopic electroretinogram was measured one week after control norbixin i.e. norbixin n.i.e. dmso i.e. dmso n.i.e. distance / optic nerve (mm) light damage and eyes were removed for histology and photoreceptor quantification. One week after BLD a- and b-wave amplitudes of In the eyes injected with norbixin we observed a partial but clear protection of photoreceptor cells and outer segments compared Bioavailability was tested in C57Bl/6 mice receiving bixin or ERG were significantly maintained in eyes injected to the contralateral eyes of the same mice or to the dmso-injected eyes. In the central retina, 4 to 6 rows of photoreceptors were norbixin, 50 mg/kg per os (in oil/DMSO 9:1) or 5 mg/kg with norbixin (i.e.) compared with non injected still present one week after BLD compared to 1-2 rows for dmso or norbixin non-injected eyes . intraperitoneally (in DMSO/tetraglycol/water 1:2:7) (N=4). eyes (n.i.e.) or dmso injected eyes. *: norbixin i.e. compared to norbixin n.i.e.; # : norbixin i.e. compared to dmso i.e. For statistical analyses, one-way ANOVA followed by Dunnett’s tests were performed. All data are presented as mean+/- s.e.m. *p < 0.05 **p < 0.01 ***p < 0.001 ****p < 0.0001 Norbixin protects the rat retina against blue light induced phototoxicity 12 CONCLUSIONS norbixin 100 11 OS 10 norbixin 50 ONL norbixin 10 1-Bixin and norbixin are more active than xanthophylls (lutein, 9 INL 8 vehicle zeaxanthin) in vitro 7 PBN 50 2-Norbixin has a much better oral bioavailability than bixin and 6 non-induced was therefore selected for further studies GCL 5 Number of photoreceptor layers photoreceptor of Number 4 non-induced vehicle PBN 50 norbixin 10 norbixin 50 norbixin 100 3 3-Norbixin provides an efficient photoprotection when injected -3,25 -2,75 -2,25 -1,75 -1,25 -0,75 -0,25 0,25 0,75 1,25 1,75 2,25 2,75 3,25 intravitreally in Abca4-/-Rdh8-/- mice eyes distance/ optic nerve (µm) 4-Norbixin is at least as efficient as PBN by intraperitoneal One week after BLD a- and b-wave amplitudes of BLD induced a massive photoreceptor degeneration in vehicle-treated rats (4-6 rows remaining) compared to non-induced rats injections in the blue light rat model ERG were significantly maintained in eyes of (10-11 rows). At 10 and 50 mg/kg norbixin induced a partial photoreceptor protection similar to that obtained with PBN (7-9 5-Chronic oral treatment of Abca4-/-Rdh8-/- mice with norbixin is norbixin and PBN injected rats compared to the rows) whereas at 100 mg/kg the protection was very close to the non-induced control (9-10 rows compared to 10-11 rows). presently under investigation and appears to (1) reduce A2E vehicle injected group. accumulation and (2) prevent the degradation of ERG..
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