RU2806096C2 - Nutriceutical ophthalmological composition for treatment of retinal pathologies with a neovascular component - Google Patents

Abstract

FIELD: ophthalmology; therapy.

SUBSTANCE: intended for the treatment of an eye disease in a subject receiving a treatment agent that counteracts VEGF. An ophthalmic nutraceutical composition is used in the treatment of an eye disease in a subject receiving a treatment agent that counteracts VEGF. This composition contains resveratrol, vitamin C, vitamin E, zinc, copper, lutein, zeaxanthin and omega-3 fatty acids from fish oil.

EFFECT: increasing the effectiveness of treatment of eye disease in a subject receiving a treatment agent that counteracts VEGF.

21 cl, 1 dwg, 2 tbl

Description

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The present invention relates to a nutraceutical ophthalmic composition containing, in particular, resveratrol, intended for administration to a subject receiving an anti-VEGF treatment, in particular, as part of the management of retinal pathology with a neovascular component.

The present application demonstrates that co-administration of such a composition and an anti-VEGF agent has a beneficial effect on the level of VEGF production and holds promise for the treatment of these ocular diseases.

BACKGROUND OF THE ART

Across Europe, the majority of cases of blindness and severe vision loss are caused by age-related retinal diseases. Given current demographic trends, the prevalence and incidence of these diseases is expected to increase. Significant pressure on European health systems will be one of the direct economic consequences.

Today, age-related macular degeneration (AMD) affects approximately 34 million people in Europe. The number of patients with AMD is expected to increase by almost 25% by 2050 given the expected growth and aging of the population. Additionally, the current and future situation is similar for diabetes-related eye diseases, including diabetic macular edema (DMO). More than 25% of patients with diabetes are affected by diabetic eye disease (DED), representing almost 4 million people across Europe. As the prevalence of diabetes increases significantly due to demographic trends and lifestyle changes, this type of disease will also increase.

In particular, AMD is a leading cause of vision impairment in people over the age of 50. In all its forms, the disease affects approximately 8% of the French population, but its incidence increases significantly with age: it affects 1% of people aged 50 to 55 years, approximately 10% of people aged 65 to 75 years, and 25% to 30% of people over 75 years of age. If only late forms of the disease combined with loss of central vision are taken into account, these figures can be reduced by approximately half.

The disease selectively affects the macula region, i.e. the central zone of the retina, leading to progressive loss of central vision, namely the visual field suitable for reading, recognizing faces or driving, etc. This is the main cause of low vision in the elderly .

The disease begins with an early, nondegenerative phase called age-related maculopathy (ARM or "early dry stage"). This phase is characterized by the accumulation of small whitish deposits (or "soft drusen") in and around the macula. These deposits are visible with a simple examination of the fundus. This phase is usually asymptomatic, but the patient may experience distortions of straight lines (“metamorphopsia”) and blurred spots.

In the most advanced stages, symptoms appear that vary depending on the severity of the disease:

• decreased visual acuity with the need for better lighting for reading or any precise work;

• increasingly blurred central vision, changes in color perception, distortion of straight lines that appear distorted and curved;

• the appearance of a dark spot in the center of the visual field, called a scotoma, difficulty recognizing faces, visual hallucinations, or a sharp decrease in visual acuity;

• Severe damage to the second eye varies greatly between subjects: it may appear quickly, within one year, within 10 years, or not at all.

An ARM can remain stable throughout your life. However, in approximately half of the cases and under the influence of several factors, ARM develops into late degenerative forms:

- atrophic form, which is a dry form in which the macula atrophies with age and is gradually replaced by scar tissue; or

- the wet form, which is an exudative form in which small blood vessels develop under the macula. These newly formed vessels bleed easily and are therefore responsible for exudates and fundus hemorrhages. If exudative AMD is present in one eye, there is a risk of developing it in the other eye.

These two later forms are characterized by approximately equal influence. They cause permanent macular damage and loss of central vision, affecting only one eye or both eyes. Mixed forms may be observed.

Mechanistically, the wet form of AMD, called neovascular or exudative AMD, as well as diabetic retinopathy, macular edema, retinal vein occlusion (RVO), or even severe myopia lead to the growth of new abnormal vessels under the retina. These fragile vessels allow serum, which is responsible for raising the retina, and/or blood to diffuse, causing retinal hemorrhages. This form develops rapidly if left untreated, with loss of vision within weeks or even days. This process can be slowed by taking medications such as anti-VEGF drugs.

VEGF (vascular endothelial growth factor) is a vascular endothelial growth factor that plays a role in the processes of angiogenesis, i.e. the sprouting of existing vessels, and vasculogenesis, corresponding to the formation of new vessels from endothelial precursors. VEGF, and more specifically the VEGF-A isoform, binds to the surface of receptors with tyrosine kinase activity, particularly VEGF-R1 and VEGF-R2, which play a key role in angiogenesis.

Thus, since 2006, certain eye diseases such as wet AMD (or "wet AMD") have been treated with VEGF inhibitors. These treatments are also increasingly used in the treatment of other neovascular diseases such as diabetic macular edema (DMO or DME), retinal vein occlusion (RVO) or myopic choroidal neovascularization (CNV or PM).

Anti-VEGF agents include, but are not limited to, antibodies or antibody fragments directed against VEGF or recombinant proteins capable of binding to VEGF receptors. Examples include ranibizumab (eg, LUCENTIS ^® , marketed by NOVARTIS PHARMA SAS, or its biosimilars developed by SAMSUNG, INTAS or FORMYCON), bevacizumab (eg, AVASTIN ^® , marketed by ROCHE), aflibercept (eg, EYLEA ^® , marketed by BAYER SANTE), abicipar pegol (in development at ALLERGAN), conbercept (eg, the product in development at CHENGDU KANGHON BIOTECH INC), faricimab (eg, the product RG7716 in development at ROCHE), or brolucizumab (NOVARTIS).

These anti-VEGF agents are typically administered by intravitreal injection, i.e., injection through the conjunctiva and sclera directly into the cavity of the eye behind the lens, i.e., the vitreous cavity containing the vitreous humor.

This involves direct injection of medications into the eye. This method of treatment has several advantages compared to other routes of administration, in particular local:

- reduction in the amount of active ingredient;

- more effective action;

- effective targeting of the area to be treated;

- reduction of undesirable effects or reduction of passage into the systemic circulation.

However, intravitreal injection remains a technical procedure performed by specialized practitioners and requires special aseptic and antiseptic conditions. Moreover, this injection can be associated with a number of rather problematic side effects: endophthalmitis, retinal detachment, ocular hypertension/glaucoma, cataracts, inflammation, subconjunctival hemorrhage, systemic toxicity, inflammation, floaters, etc. Additionally, It is generally not recommended to inject both eyes on the same day to avoid complete loss of vision in case of problems or side effects after the injection. In reality, these injections are usually followed by a very painful period for patients, which may require them to spend several hours in complete darkness.

The dosage, in particular the dose and frequency of administration, may depend largely on the active ingredient as well as the pathology being treated, with an average number of seven injections per year.

For example, treatment of AMD with ranibizumab or bevacizumab begins with an induction phase with 1 injection per month for 3 consecutive months, followed by a maintenance phase during which patients' visual acuity is tested once a month. If the patient experiences a loss of visual acuity of more than 5 letters on the Early Treatment Diabetic Retinopathy Study (ETDRS) scale or equivalent to one line on the Snellen scale, the drug should be reintroduced. The interval between 2 doses should be at least 1 month.

For DMO, monthly injections of ranibizumab or bevacizumab are recommended until visual acuity stabilizes at three consecutive monthly assessments as part of treatment.

For RVO, monthly injections of ranibizumab or bevacizumab are recommended until visual acuity stabilizes. Failure to improve visual acuity after 3 consecutive monthly injections is grounds for discontinuation of treatment.

If aflibercept is present, the protocol is slightly different.

For AMD, at the beginning of treatment, as with other anti-VEGF agents, aflibercept is injected once a month for 3 consecutive months, then one injection is given every 2 months with no follow-up visits between injections. After the first 12 months of treatment, the interval between 2 injections may be increased depending on the results of the study of visual function and anatomical structures. In this case, the follow-up schedule should be determined by the treating physician, and follow-up visits may be more frequent than scheduled injections.

Regarding RVO, the recommended dose is 2 mg of aflibercept, which corresponds to 50 μl. After the first injection, treatment is carried out monthly. The interval between 2 injections must be at least 1 month. If after the first three injections there is no improvement in visual and anatomical parameters, then further treatment is not recommended. Monthly treatment is continued until stable visual and anatomical results are achieved at three monthly assessments. The need for further treatment can then be re-evaluated. If necessary, treatment can be continued with a gradual increase in the interval between injections to maintain the response from visual function and anatomical structures.

In parallel with these treatments using anti-VEGF agents administered by intravitreal injection, other techniques are used to destroy newly formed vessels, in particular photocoagulation (thermal destruction of abnormal vessels) and photodynamic therapy. The latter technique involves the intravenous injection of a photosensitive product (eg, verteporfin) that becomes toxic when exposed to red light applied locally with a laser.

Prior to these harsh treatments, which are given in the later stages of the disease, research has been conducted in an attempt to find earlier and “easier” treatments to prevent the onset of symptoms and delay the progression of the disease.

Thus, the study called AREDS 1 (Age-Related Eye Disease Study Research Group. A Randomized, Placebo-Controlled, Clinical Trial of High-Dose Supplementation With Vitamins C and E, Beta Carotene, and Zinc for Age-Related Macular Degeneration and Vision Loss: AREDS Report No. 8 and 9. Arch Ophthalmol. 2001) and AREDS 2 (Lutein + zeaxanthin and omega-3 fatty acids for age-related macular degeneration: the Age-Related Eye Disease Study 2 (AREDS2) randomized clinical trial JAMA 2013 May 15;309(19):2005-15. doi: 10.1001/jama.2013.4997) was conducted to demonstrate the beneficial effect of dietary supplements on eye diseases.

These studies have identified antioxidant-based dietary supplements as potentially beneficial in the treatment of eye diseases, particularly age-related diseases, especially AMD.

In practice, the AREDS 1 study recommends daily intake of antioxidants such as micronutrients and vitamins, as follows:

These supplements may reduce the risk of vision loss in patients with advanced AMD.

The AREDS 2 study was conducted to illustrate the effects of supplementation with lutein, zeaxanthin (as a substitute for beta-carotene), and omega-3 fatty acids. This study recommended eliminating vitamin A and reported benefits from supplementing daily intake of the following:

Therefore, based on the results of the AREDS studies, daily dietary supplementation is recommended for people at high risk of developing AMD, especially patients with ARM in one or both eyes, as well as patients with advanced AMD that is still affecting only one eye. However, these studies did not demonstrate a beneficial effect of these formulations on disease progression in patients with advanced AMD or on vision loss.

The NAT 2 study (Merle et al ., Invest Ophthalmol Vis Sci. 2014 Mar 28;55(3):2010-9. doi: 10.1167/iovs.14-13916), including 300 patients over a three-year period, demonstrated 68% reducing the risk of developing neovascular AMD after a significant increase in the amount of DHA in cells. Thus, this study is the first to examine the potential of long-term oral DHA supplementation to prevent the development of AMD.

Finally, WO 2016/151269 describes a dietary supplement that combines vitamins, trace elements, carotenoids, omega-3 fatty acids and resveratrol, and its use for the treatment and/or prevention of eye diseases. In particular, this document reports that the combination of various listed compounds has a differential and beneficial effect on the expression and activity of VEGF or its receptors (VEGF-R).

However, there is a clear need to develop new solutions to improve the control of ocular diseases, in particular retinal diseases with a neovascular component, meeting, in particular, the following expectations: increasing the intervals between intravitreal injections of an anti-VEGF agent, reducing associated side effects, reducing the costs of public health and improving the quality of life of patients.

BRIEF DESCRIPTION OF THE INVENTION

Applicant has illustrated that the administration of a dietary supplement will enhance, prolong or even improve the effects of currently used anti-VEGF treatments, particularly treatments administered by intravitreal injection, in patients receiving such treatments.

According to its first aspect, the present invention relates to a nutraceutical ophthalmic composition containing resveratrol, and the administration of such composition to a subject receiving an anti-VEGF treatment, in particular undergoing treatment with an anti-VEGF agent.

Within the scope of the present invention, a “nutraceutical composition” is defined as a composition the intended purpose of which is to supplement the normal diet and to provide a concentrated source of nutrients or other substances that have a nutritional or physiological effect alone or in combination. It advantageously provides a dietary supplement.

Within the scope of the present invention, a composition with properties beneficial to eye health is referred to as an "ophthalmic composition". However, this does not exclude the possibility that such a composition may have a beneficial effect on other organs of humans or animals.

Resveratrol is a polyphenolic compound derived from stilbene, which has two isomers, but the trans form is predominantly active. Therefore, in the remainder of this specification, the term “resveratrol” may be used instead of trans -resveratrol.

Resveratrol is a polyphenol found in abundant amounts in certain fruits, particularly grapes, oilseeds and wine. There are currently several commercial sources of resveratrol that can be used to prepare the composition within the scope of the present invention:

• Vitis vinifera :

• grape skins

• grape seed

• Polygonum cuspidatum:

• obtained by fermentation

Preferred sources include:

- source of resveratrol Vitis vinifera , grape skin: for example, natural extract with a content of 5%;

- 99% synthetic resveratrol (CAS number: 501–36–0).

Resveratrol is preferably present in an amount (daily dose) that is greater than or equal to 1 mg, or even 2 mg, 5 mg, 10 mg, 20 mg, 30 mg, 50 mg, 60 mg, or even 100 mg or 200 mg. According to a specific embodiment, the daily amount thereof is greater than or equal to 20 mg or even greater than 20 mg, greater than or equal to 30 mg or even greater than 30 mg, greater than or equal to 50 mg, greater than or equal to 60 mg or even greater than or equal to 100 mg. Advantageously it is less than or equal to 1 g, preferably less than or equal to 500 mg, particularly preferably less than or equal to 200 mg. According to a specific embodiment, a composition within the scope of the present invention contains a daily dose of resveratrol equal to 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg or 200 mg.

According to an advantageous embodiment, the composition additionally contains vitamins, microelements, carotenoids and omega-3 fatty acids. The ingredients of vitamins, trace elements, omega-3 fatty acids and carotenoids, known for their antioxidant properties, are even more preferentially selected in accordance with the recommendations of the AREDS studies.

In a specific embodiment, the composition contains at least one vitamin, preferably a combination of vitamins. It predominantly contains vitamin C and/or vitamin E.

These may also include the following vitamins:

- nicotinamide, niacinamide or vitamin B3 or PP;

- pyridoxine, for example in the form of pyridoxine-HCl, or vitamin B6;

- riboflavin or vitamin B2;

- thiamine, for example in the form of thiamine nitrate, or vitamin B1;

- cyanocobalamin or vitamin B12;

- folic acid or vitamin B9;

- vitamin B5;

- vitamin B8;

- vitamin D.

Vitamin C, available in the form of ascorbic acid or calcium ascorbate, is preferably present in an amount (daily dose) of 50 mg to 500 mg, for example 240 mg or 120 mg.

Vitamin E or alpha-tocopherol, for example, available in the form of 67% oil, is preferably present in an amount (daily dose) of from 10 mg to 500 mg, for example equal to 30 mg.

In another embodiment, the following daily amounts, expressed in mg, may be considered for the following vitamins:

- vitamin B3 or PP: from 1 to 100, for example 18;

- vitamin B6: from 1 to 10, for example 2;

- vitamin B2: from 1 to 10, for example 1.6;

- vitamin B1: from 1 to 10, for example 1.4;

- vitamin B12: from 0.0005 to 0.01, for example 0.001;

- vitamin B9: from 0.1 to 1, for example 0.2.

In a specific embodiment, the composition does not contain vitamin A or beta-carotene.

According to a specific embodiment, the composition contains at least one micronutrient, preferably a combination of micronutrients. It predominantly contains zinc (Zn) and/or copper (Cu). It can also be the following microelements:

- manganese, for example anhydrous manganese sulfate;

- selenium, for example sodium selenite;

- magnesium, for example magnesium oxide.

Zinc (Zn), available as zinc oxide or zinc sulfate (monohydrate), is advantageously present in an amount (daily dose) of 5 mg to 100 mg, for example 12.5 mg.

Copper (Cu), available in the form of copper sulfate, copper monohydrate or anhydrous copper, is preferably present in an amount (daily dose) of 0.2 mg to 10 mg, for example in an amount of 1 mg.

In another embodiment, the following daily amounts, expressed in mg, may be considered for the following micronutrients:

- manganese: from 1 mg to 10 mg, for example 1 mg;

- selenium: from 0.01 mg to 0.1 mg, for example 0.025 mg; And

- magnesium: from 1 mg to 50 mg, for example 10 mg.

According to a specific embodiment, the composition contains at least one carotenoid, preferably a combination of carotenoids. It predominantly contains lutein and/or zeaxanthin. These may also include the following carotenoids:

- mesozeaxanthin;

- lycopene;

- astaxanthin.

Lutein, available in the form of 20% lutein by weight, is preferably present in an amount (daily dose) of 2 mg to 50 mg, for example 10 mg or 20 mg.

Zeaxanthin, available in the form of 5% or 14% zeaxanthin by weight, is preferably present in an amount (daily dose) of 0.5 mg to 10 mg, for example equal to 2 mg or 4 mg.

In a specific embodiment, the composition contains at least one omega-3 polyunsaturated fatty acid, preferably a combination of omega-3 fatty acids. It predominantly contains eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA). These may also include the following omega-3 fatty acids:

- alpha-linolenic acid (ALA);

- docosapentaenoic acid (DPA).

In a specific embodiment, the composition contains at least one omega-3 polyunsaturated fatty acid, preferably a combination of omega-3 fatty acids.

The omega-3 fatty acids used in such a composition can be obtained from fish oil rich in omega-3 fatty acids or from a vegetable oil, such as flaxseed oil. These fatty acids are predominantly present in amounts (daily dosage) of 100 mg to 1000 mg. More generally, fish oil may be present in daily amounts of 400 mg to 1000 mg or even 800 mg to 1000 mg.

In this case, EPA is preferably present in an amount (daily dose) of 100 mg to 1000 mg, for example 172 mg.

In this case, DHA is preferably present in an amount (daily dose) of 100 mg to 1000 mg, eg 366 mg.

In certain embodiments, a composition useful within the scope of the present invention may contain other active ingredients, such as:

- one or more other polyphenols, such as epigallocatechin-3-gallate (EGCG);

- glutathione, for example, from 1 mg to 10 mg per day or even 2 mg per day;

- anthocyanoside(anthocyanosides);

- hydroxytyrosol.

The content of each of these ingredients is easily determined and adjusted by one skilled in the art.

Thus, and according to one embodiment, the composition contains or consists of the following ingredients:

- vitamin C;

- vitamin E;

- possibly vitamin D;

- zinc;

- copper;

- lutein;

- zeaxanthin;

- omega-3 fatty acids from fish oil, in particular EPA and DHA;

- resveratrol.

According to another embodiment, the composition contains or consists of the following ingredients per daily dose:

- from 50 mg to 500 mg of vitamin C, preferably 240 mg, more preferably 120 mg;

- from 10 mg to 500 mg of vitamin E, preferably 30 mg;

- from 5 mg to 100 mg of zinc, preferably 12.5 mg;

- from 0.2 mg to 10 mg of copper, preferably 1 mg;

- from 2 mg to 50 mg of lutein, preferably 20 mg;

- from 0.5 mg to 10 mg of zeaxanthin, preferably 4 mg;

- from 400 mg to 1000 mg of fish oil, especially preferably from 800 to 1000 mg;

- resveratrol, preferably in an amount greater than or equal to 20 mg, advantageously greater than 30 mg, especially advantageously greater than 50 mg, very advantageously in an amount greater than or equal to 60 mg;

- optional vitamin D, preferably 5 mcg.

According to a preferred embodiment, the composition contains or consists of the following ingredients per daily dose:

- 120 mg vitamin C;

- 30 mg vitamin E;

- 12.5 mg zinc;

- 1 mg copper;

- 20 mg of lutein;

- 4 mg zeaxanthin;

- 950 mg fish oil, mainly containing 366 mg DHA and 172 mg EPA;

- resveratrol, preferably in an amount that is greater than or equal to 50 mg, very preferably in an amount that is greater than or equal to 60 mg.

Alternatively, the composition contains or consists of the following:

- 240 mg vitamin C;

- 30 mg vitamin E;

- 12.5 mg zinc;

- 1 mg copper;

- 10 mg of lutein;

- 2 mg zeaxanthin;

- 950 mg fish oil;

- resveratrol, preferably in an amount that is greater than or equal to 20 mg, very preferably in an amount that is greater than or equal to 30 mg.

In a specific embodiment, the composition does not contain at least one of the following ingredients:

- saffron;

- a metal chelating agent such as EDTA or phytic acid;

- nucleotide;

- pycnogenol;

- Ginkgo biloba extract.

According to this embodiment, the composition used does not combine resveratrol with saffron, or resveratrol with a metal chelating agent and one or more nucleotides, or resveratrol and pycnogenol and/or Ginkgo biloba extract.

The composition within the scope of the present invention may be present in the form of a liquid, solution, suspension, paste or gel.

It is advantageously presented in solid form, such as powder, tablets, including effervescent tablets, pills, capsules or lozenges, prepared in a conventional manner using suitable additives such as binders, fillers, lubricants, disintegrants or wetting agents. The tablets may optionally be coated in a manner known to one skilled in the art using sugars, films or enteric coatings.

Alternatively, other delivery systems such as soft gelatin or gelatin-based capsule formulations or nanotechnology-derived formulations such as nanodispersions, nanoemulsions or nanocapsules can be used.

Such a composition may further contain one or more additives such as colorants, pigments or flavoring agents, etc.

In a specific embodiment, the composition is presented in the form of capsules defining a predetermined volume in which the composition is contained. The capsule breaks down in the digestive system to release the active ingredients and allow them to be absorbed by the body.

In this case, traditional additives are:

- glycerol, for example glycerol monostearate; and/or

- soybean oil; and/or

- beeswax; and/or

- soy lecithin.

When appropriate, these additives are added to the composition due to their ability to improve, for example, its flow properties or homogenization.

Additionally, and as is known to one skilled in the art, the capsule shell can be made from the following ingredients:

- gelatin; and/or

- iron oxide red; and/or

- iron oxide black; and/or

- sorbitol; and/or

- glycerin.

As one skilled in the art knows, the distribution in this type of packaging is a compromise between:

- a limited number of doses to ensure adequate adherence to the treatment regimen;

- an appropriate dose volume that allows all ingredients to be combined and is easy to swallow.

Such a composition is advantageously administered daily, i.e. every day.

According to an advantageous embodiment, the composition is formulated in 2 capsules, each containing half the ingredients listed above.

In a first alternative, all of these ingredients are mixed and administered as a single composition. Alternatively, these ingredients can be combined in various ways to be formulated and/or administered simultaneously or spaced apart.

Typically, the composition used within the scope of the present invention is administered orally, preferably in the form of 2 daily doses. According to a specific embodiment, the composition is presented in the form of 2 capsules, which can be taken with a small amount of water. They can be taken at the same time, for example at the beginning of the main meal or during the main meal.

In view of the effect illustrated in the examples, such a composition is intended for administration to a subject receiving an anti-VEGF treatment.

In another aspect, an anti-VEGF treatment agent, as will be described in detail below, is intended to be administered to a subject taking a composition as defined above, preferably orally, even more preferably orally, on a daily basis.

According to a specific embodiment and within the scope of the present invention, the anti-VEGF treatment corresponds to the administration of an anti-VEGF agent, more specifically an agent that inhibits the VEGF signaling pathway.

The anti-VEGF agent is preferably selected from an antibody, an antibody fragment, a recombinant protein, a fusion protein, and a mixture thereof.

In a specific embodiment, the anti-VEGF agent is selected from the group consisting of ranibizumab, bevacizumab, aflibercept, abicipar pegol, conbercept, faricimab, brolucizumab, biosimilars thereof, and mixtures thereof.

For the purposes of the present invention, a biosimilar in the generally accepted sense of the term refers to a drug product that is similar to a biological substance obtained from a living cell or organism, called the original drug product. This biosimilar medicinal product must have similar physicochemical and biological properties, the same pharmaceutical substance and the same pharmaceutical form compared to the original medicinal product, as well as equivalent efficacy and safety.

Ranibizumab is a humanized monoclonal antibody fragment against VEGF-A.

Bevacizumab is a recombinant humanized monoclonal antibody against VEGF.

Aflibercept is a recombinant fusion protein corresponding to the extracellular domains of human VEGF receptors types 1 and 2 (VEGF-R1 and VEGF-R2) fused to a crystallizable fragment (Fc) of human IgG1 immunoglobulin.

Abicipar pegol is an ankyrin repeat protein that binds to all VEGF-A isoforms. Its small size (34 kDa) suggests that it has a short half-life in the eye, but it acquires the pharmacokinetic characteristics of a larger protein, particularly a longer half-life, through PEGylation, i.e., the addition of polyethylene glycol molecules.

Conbercept is similar to aflibercept in that it is a recombinant fusion protein corresponding to the key extracellular domains of VEGF receptors types 1 and 2 and human IgG Fc. However, conbercept also contains a fourth domain of the VEGF-2 receptor.

Faricimab is a bispecific monoclonal antibody to angiopoietin 2 and VEGF.

Brolucizumab (or RTH258, Novartis) is a single-chain antibody that provides much higher levels of antibody in the eye than currently approved treatments. This antibody has very high affinity for all isoforms of VEGF-A and is expected to increase the interval between intravitreal injections.

Typically, these anti-VEGF agents are administered by intravitreal injection. However, given the enhancement of their effect in the presence of the composition described above, the possibility of their topical application, mainly on the surface of the eye, can be considered.

The combination of these two treatments (a nutraceutical ophthalmic composition for oral administration and an anti-VEGF treatment administered by intravitreal injection or topically) is advantageous for the treatment of ocular diseases, preferably retinal diseases with a neovascular component, even more advantageously selected from the group consisting of :

- age-related macular degeneration, predominantly wet form of AMD;

- diabetic macular edema (DMO);

- retinal vein occlusion (RVO);

- myopic choroidal neovascularization (mCNV);

- neovascular glaucoma (NVG) and

- proliferative diabetic retinopathy (PDR).

According to another aspect, the present invention relates to a combination product or combination preparation containing a nutraceutical composition and an anti-VEGF treatment agent described above for simultaneous, separate or sequential use in the treatment of an eye disease, preferably a retinal disease with a neovascular component, even more advantageously selected from group consisting of:

- age-related macular degeneration, predominantly wet form of AMD;

- diabetic macular edema (DMO);

- retinal vein occlusion (RVO);

- myopic choroidal neovascularization (mCNV);

- neovascular glaucoma (NVG) and

- proliferative diabetic retinopathy (PDR).

As already mentioned, in this case the nutraceutical composition is advantageously administered orally, even more advantageously daily, for example in the form of 2 capsules taken at the beginning or in the middle of the main meal.

At the same time, an anti-VEGF treatment agent is administered by intravitreal injection or topically. Intravitreal injection remains the preferred route of administration, but recent developments allow consideration of topical application, preferably by application to the ocular surface.

Indeed, this combination has been demonstrated to reduce the dosage of anti-VEGF treatments in particular. Thus, it is possible to reduce the doses used or, advantageously, to increase the interval between administrations (or reduce the frequency of administration), which are usually separated by intervals of 1-3 months.

In other words, this combination product allows for a reduction in the dose and/or frequency of administration of the anti-VEGF treatment compared to the dose and/or frequency administered to a subject not receiving the nutraceutical composition.

In another aspect, the interval between two administrations of the anti-VEGF treatment may be greater than 30 days, preferably greater than 45 days, preferably greater than 60 days, or even 75 days, more advantageously greater than 90 days, compared to the currently used dosage.

Thus, and according to another aspect, the interval between two administrations of the anti-VEGF treatment is greater than 30 days, preferably greater than 45 days, preferably greater than 60 days, or even 75 days, more preferably greater than 90 days.

In this way, the potential toxicity of anti-VEGF treatments, in particular their side effects, can be reduced. The interval between their administrations, requiring specialist intervention, is also of interest from an economic point of view.

Exemplary Embodiments of the Present Invention

The present invention and the advantages resulting therefrom will become more apparent from the following figures and examples, which are given to illustrate the present invention and without limitation.

DESCRIPTION OF FIGURES

Figure 1 compares the dynamics of changes in VEGF levels after administration of AVASTIN ^® in incubated undifferentiated (pathological) retinal cells (A) in the presence (lower curve) and in the absence (upper curve) of formula A or (B) in the presence (lower curve) and in absence (upper curve) of formula B.

A. Protocol

Cell culture

Undifferentiated human ARPE-19 retinal cells (ATCC® CRL-2302™) were cultured in vitro in DMEM (Dulbecco's modified Eagle's medium/F12 nutrient mixture (1:1, Gibco, Paisley, UK) supplemented with 10% fetal calf serum (FCS). , Dominique DUTSCHER, South American origin, lot no. S15197S1810), 1% PSA (cocktail of antibiotics: penicillin, streptomycin and amphotericin B, respectively at 10,000 units/ml: 10 mg/ml: 25 μg/ml, PAN Biotech) and 15 mM HEPES (pH 7.4, Gibco) Cells are grown in an atmosphere of 37°C and 5% CO _2. These cells can become similar to diseased or pathological retinal cells.

Types of processing

Cells were grown and seeded in 24-well culture dishes at 60,000 cells per well in DMEM/F12, 10% SVF, and 1% PSA. 24 hours after plating, growth medium was removed and replaced with restrictive growth medium DMEM/F12 with 1% SVF and 1% PSA without phenol red. After another 24 hours, cells were placed in the presence or absence of formula A (with resveratrol) or formula B (without resveratrol).

The composition of Formulas A and B is described below.

Table 1. Composition of Formula A (with resveratrol)

Please note that one capsule contains 690 mg of the drug, and one daily dose of 2 capsules is recommended.

Table 2. Composition of Formula B (no resveratrol)

Please note that one capsule contains 760 mg of the drug and it is recommended to take only 1 capsule per day.

Formulas A and B are taken at a weight of 0.115 g in 1 ml of DMSO (dimethyl sulfoxide) to give a resveratrol equivalent of 10 mM for Formula A and 0 mM for Formula B. Indicators of fish oil, vitamins C and E, zinc, copper, lutein and zeaxanthin are considered equivalent in these 2 drugs. From this solution, dilutions are made directly in DMEM/F12 growth medium with 1% SVF, 1% PSA without phenol red to achieve the 10 µM and 20 µM levels required for the experiment.

After 24 hours of pre-incubation, cells are treated with 100 ng/ml AVASTIN ^® (± formula A or B).

ELISA (enzyme-linked immunosorbent assay)

At the time of treatment (t0) with 100 ng/ml Avastin ^® , and 24 hours, 48 hours and 72 hours after treatment, cell culture supernatant is collected for analysis of the amount of VEGF-A released. Each ELISA was performed according to the manufacturer's exact instructions (human VEGF-A ELISA kit BMS277, Affymetrix, eBioscience).

Crystal violet

The cell layer is analyzed using crystal violet to determine cell viability. The relationship between the levels of VEGF-A in the supernatant and the corresponding cell layer can be determined.

The cell layer is gently rinsed twice with 1X PBS buffer. When excess liquid is removed, the cell layer is fixed on the substrate using absolute ethanol. After incubation for a period of at least 30 minutes, absolute ethanol is removed. The cells are brought into contact with a crystal violet staining solution for 20 minutes at room temperature and under vibration (25 vibrations per minute).

The crystal violet solution is removed and the wells are rinsed 4 times with water to remove excess dye. The plates are then dried at room temperature (for 16 to 24 hours). The stained cells are then resuspended in 33% acetic acid, the plates are placed under vibration conditions, and the optical density can be read spectrometrically at a wavelength of 570 nm (OD ₅₇₀ ).

B. Results

It can be observed that the presence of Formula A allows the amount of VEGF produced to be limited and also significantly delays the increase in this level after treatment with an anti-VEGF agent, allowing for longer intervals between injections of the anti-VEGF agent.

In contrast, Formula B results in a reduction in the amount of VEGF produced but does not delay the recovery of VEFG production in retinal cells treated with an anti-VEGF agent.

This unexpected difference is due to the presence of high resveratrol content in Formula A.

Claims (28)

1. The use of an ophthalmic nutraceutical composition containing resveratrol in an amount of more than 20 mg in the treatment of an eye disease in a subject receiving an anti-VEGF treatment agent, characterized in that the composition contains: - vitamin C in amounts from 50 mg to 500 mg; - vitamin E in amounts from 10 mg to 500 mg; - zinc in an amount from 5 mg to 100 mg; - copper in amounts from 0.2 mg to 10 mg; - lutein in amounts from 2 mg to 50 mg; - zeaxanthin in an amount from 0.5 mg to 10 mg; - omega-3 fatty acids from fish oil in amounts from 100 mg to 1000 mg. 2. Use according to claim 1, characterized in that the composition contains vitamin C in an amount of 120 mg. 3. Use according to claim 1, characterized in that the composition contains vitamin E in an amount of 30 mg. 4. Application according to claim 1, characterized in that the composition contains zinc in an amount of 12.5 mg. 5. Application according to claim 1, characterized in that the composition contains copper in an amount of 1 mg. 6. Use according to claim 1, characterized in that the composition contains lutein in an amount of 20 mg. 7. Use according to claim 1, characterized in that the composition contains zeaxanthin in an amount of 4 mg. 8. Use according to claim 1, characterized in that the composition contains omega-3 fatty acids from fish oil in an amount of from 800 mg to 1000 mg. 9. Use according to claim 1, characterized in that the omega-3 fatty acids from fish oil are selected from the group consisting of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). 10. Use according to claim 1, characterized in that the composition contains resveratrol in an amount that is greater than or equal to 30 mg, preferably greater than 30 mg, even more preferably greater than or equal to 50 mg. 11. Use according to any of the previous paragraphs, characterized in that the composition is administered orally. 12. Use according to any of the previous paragraphs, characterized in that the composition is administered daily. 13. Use according to any of the previous claims, characterized in that the anti-VEGF treatment corresponds to the administration of an inhibitor of the VEGF signaling pathway, preferably selected from the following group: antibody, antibody fragment, recombinant protein, fusion protein and mixtures thereof. 14. Use according to any of the previous paragraphs, characterized in that the inhibitor is selected from the following group: ranibizumab, bevacizumab, aflibercept, abicipar pegol, conbercept, faricimab, brolucizumab and mixtures thereof. 15. Use according to any of the previous claims, characterized in that the anti-VEGF treatment agent is administered by intravitreal injection or topically. 16. The use of any of the preceding claims, wherein the eye disease is selected from the group consisting of age-related macular degeneration, diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma and proliferative diabetic retinopathy. 17. Application according to any one of paragraphs. 1-10 with an anti-VEGF agent, preferably specified in paragraphs. 13 and 14, in the treatment of an eye disease preferably selected from the group consisting of age-related macular degeneration, diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma and proliferative diabetic retinopathy. 18. Use according to claim 17, characterized in that the composition is administered orally, preferably daily. 19. Use according to claim 17 or 18, characterized in that the anti-VEGF agent is administered by intravitreal injection or topically. 20. Application according to any one of paragraphs. 17-19, wherein the dose and/or frequency of administration of the anti-VEGF agent is reduced compared to the dose and/or frequency administered to a subject not receiving the composition. 21. Use according to claim 20, characterized in that the interval between two administrations of the anti-VEGF agent is more than 30 days, preferably more than 45 days, preferably more than 60 days or even more than 75 days, more preferably more than 90 days, compared to the interval between two administrations of an anti-VEGF agent in a subject not receiving the composition.