Composition for use in the treatment and / or prevention of degenerative eye diseases

Abstract

1. An orally administered composition for use in the prevention and / or treatment of degenerative eye diseases, comprising a lysate of probiotic microorganisms in the form of a dry powder of the genera Bacillus, Lactobacillus, Streptococcus, Saccharomyces, and Bifidobacterium.

Description

[Technical Field]

[0001] The present invention describes an oral administration composition for preventing and treating ocular degenerative diseases, characterized by comprising a lysate of probiotic microorganisms in the form of a dry powder.Therefore, the present invention is within the technical field of treatment for ocular degenerative diseases, particularly age-related macular degeneration (AMD). [Background technology]

[0002] With improvements in medical care and the modernization of society, the average life expectancy of the population is increasing. This is accompanied by an increase in age-related diseases, and it is becoming increasingly clear that aging is an important factor in the development of degenerative diseases. These diseases not only cause a decline in central vision, but also include those that cause a decline in total vision, such as inherited degenerative diseases of the retina. Known diseases related to ocular degeneration include age-related macular degeneration (AMD), glaucoma, and dry eye. All of these have a negative impact on patients' quality of life and medical costs. Elderly people are at highest risk of developing these diseases, but the underlying mechanisms of these diseases are often unknown.

[0003] AMD is the leading cause of blindness in people over 65 in industrialized countries. It affects 30 to 50 million people and is expected to increase ten-fold by 2040, despite the introduction of new treatments and prevention strategies. Its prevalence increases with age and shares features with other diseases, such as Alzheimer's disease, due to the presence of abnormal extracellular deposits associated with neurodegeneration, drusen, and plaques, respectively.

[0004] Macular degeneration is an eye disease, most commonly the atrophic or dry type. With age, the retina thins, the cells in the macula deteriorate, and viable cells are replaced by nutrient cells. As the nutrient cells expand, they reach the center of the retina, causing a slow and gradual loss of vision. Once dry macular degeneration progresses, no treatment can prevent vision loss.

[0005] While the exact pathogenesis of AMD is unknown, exposure to risk factors such as age, genetics, lifestyle, environmental factors, and diet is thought to influence its progression. The onset of AMD is thought to be caused by exposure to environmental factors at an advanced age that induce high levels of oxidative stress, which damages the macula. This damage then triggers inflammation, leading to vision loss, creating a vicious cycle. As a defense mechanism, the eye has an antioxidant system, primarily composed of vitamins, enzymes, and carotenoids, which work synergistically to protect against the destruction of free radicals. Vitamins C and E and the carotenoids lutein and zeaxanthin are the most important. Furthermore, the retina contains many easily oxidized polyunsaturated fatty acids and photosensitive compounds. These are thought to contribute to the permeability, thickness, and fluidity of photoreceptor membranes, and their deficiency is linked to altered retinal function. With aging, the antioxidant system deteriorates due to damage to cellular structures, increasing the generation of free radicals and leading to lipid peroxidation. These peroxides interact with proteins, nucleic acids, and other components, affecting ocular structure and function. Degeneration of photoreceptor cells and accumulation of oxidative metabolites can lead to drusen formation and changes in the retinal pigment epithelium (RPE), potentially leading to AMD.

[0006] Since increased oxidative damage is one of the main contributing factors, it is logical to assume that consuming antioxidants can help prevent or slow the progression of AMD. Diet is the primary source of antioxidants, providing vitamins, carotenoids, essential fatty acids, and trace elements, and healthy individuals typically consume sufficient amounts. However, diet alone is not sufficient to provide the amount needed to protect the eyes. To restore balance, antioxidant supplements have been proposed as a treatment to slow the progression of AMD. One such study, the Age-Related Eye Disease Studies (AREDS) 1 and 2, showed that dietary supplementation with antioxidants and micronutrients effectively reduced the progression to advanced AMD by 28% over a five-year period in patients aged 55 years or older (7). This is currently the most common preliminary registration for early and intermediate AMD.

[0007] There are multiple studies and publications on techniques and methods for treating macular degeneration. Currently, one of the treatments for AMD patients is intravitreal injection of anti-VEGF drugs, which are anti-vascular endothelial growth factor drugs. However, while this treatment is highly effective, it has technical limitations, such as the need for specialist consultation and the painful and cumbersome nature of the procedure.

[0008] In other degenerative diseases, such as glaucoma, the mechanism that causes vision loss appears to be the result of optic nerve damage. Most treatments aim to control intraocular pressure with medication or surgery, and if treatment is ineffective, surgery is performed. In either case, the damage caused is irreversible.

[0009] Additionally, in inherited diseases such as Stargardt disease, there is an underlying inflammatory problem that, if treated, can improve prognosis or slow the progression of the disease.

[0010] Therefore, in order to address the technical challenges of current treatments, the field is actively working to develop treatments and compositions that can be administered orally to patients.Recent studies have shown that interactions between the microbiota and the eye may influence the progression of disease.

[0011] As evidence, the following patent documents are cited:

[0012] International Patent Application WO2022125925A describes a method for treating dry age-related macular degeneration (dry AMD), comprising administering a therapeutically effective amount of a compound or pharmaceutical composition to a subject in need thereof. The method describes a pharmaceutical formulation.

[0013] U.S. Patent No. 8,603,522 B2 describes a daily nutritional or dietary supplement composition that enhances and promotes retinal health through the prevention, stabilization, reversal, and / or treatment of early age-related macular degeneration. The ingredients of the daily nutritional or dietary supplement composition include vitamin C, vitamin E, lutein, zinc, and copper. The composition is preferably provided in the form of a tablet suitable for oral ingestion.

[0014] International Patent Application WO2021224679A1 relates, in part, to a method for preventing or treating macular degeneration in a subject by co-administering the enzyme superoxide dismutase and probiotic Bacillus spores, particularly mutant strains of Bacillus amyloliquefaciens GF423 or GF424. The present invention also provides pharmaceutical and / or food compositions comprising the enzyme superoxide dismutase and probiotic Bacillus spores.

[0015] It is also worth mentioning the inventors' patent applications, Spanish Patent Applications P201930242 and P201930280, the contents of which are incorporated herein by reference, which describe a composition for modulating the human intestinal microbiome obtained from a lysate of probiotic microorganisms and a method for obtaining it, as well as a food supplement comprising said composition, which are useful for the prevention and treatment of disorders caused or at least promoted by intestinal dysbiosis of the microbiota in humans.

[0016] However, none of the literature in the current state of the art discloses a composition consisting of dried bacterial lysate that allows for the reduction and prevention of macular degeneration, as described herein. Summary of the Invention [Problem to be solved by the invention]

[0017] The present invention therefore aims to solve the problems of the prior art by using an orally administered composition obtained from a lysate of a probiotic microorganism for use in the treatment of macular degeneration in humans. [Means for solving the problem]

[0018] Brief description of the invention In a first aspect, the present invention relates to an orally administered composition for use in the prevention and / or treatment of degenerative eye diseases, characterized in that it contains probiotic lysates in the form of a dry powder in the following ranges of percentages by weight relative to the total amount of probiotic microorganisms: - Bacterial lysate of Bacillus spp. lysates) at 15% and 20%, - 15% and 35% of bacterial lysates of Lactobacillus spp. - 1.5% and 8% of bacterial lysates of Streptococcus spp. - 45% and 60% of bacterial lysates of Saccharomyces spp., and - 1.5% and 8% of bacterial lysates of Bifidobacterium spp.

[0019] In a second aspect, the present invention relates to the use of the subject composition of the present invention as a food supplement for preventing and treating ocular degenerative diseases. In a preferred aspect, the present invention relates to the use of the subject composition of the present invention as a food supplement for preventing age-related macular degeneration.

[0020] In a third aspect, the present invention relates to a composition as defined herein for use as a medicament.

[0021] In a fourth aspect, the present invention relates to a composition as defined herein for use in the treatment of macular degeneration.

[0022] In a fifth aspect, the present invention relates to a pharmaceutical composition comprising an effective pharmaceutical amount of a composition according to the first aspect of the invention, and a pharmaceutically acceptable excipient.

[0023] To complement this specification and to facilitate a better understanding of the present invention, attached as an integral part hereof are a series of figures which, by way of example and not limitation, illustrate the experimental results of the pilot study described in Example 3 herein. [Brief explanation of the drawings]

[0024] [Figure 1] Figure 1 shows the progression of atrophy in patient C1 after 12 and 18 months. [Figure 2] Figure 2 shows an example of the trend of atrophy changes in patient C1. [Figure 3] Figure 3 shows an example of atrophic changes in C6 in an untreated patient. [Figure 4] Figure 4 shows the change in area in SQRT, BSL, before and after 12 months of treatment in 9 patients treated with the postbiotic composition of the present invention (26.58% reduction). Each line represents a patient's eye. [Figure 5] Figure 5 shows the change in SQRT area before and after 12 months of untreated BSL in nine patients without supplementation (4.5% increase). Each line represents a patient's eye. DETAILED DESCRIPTION OF THE INVENTION

[0025] As indicated in the previous section, the present inventors have recognized a need to develop compositions that are easy to administer and non-toxic to patients suffering from degenerative eye diseases.

[0026] Based on this, they developed an orally administered composition containing a microbial lysate and other essential components of the composition, which proved surprisingly beneficial to patients suffering from degenerative eye diseases.

[0027] Thus, in a first aspect, the present invention provides an orally administered composition for preventing and treating degenerative eye diseases, characterized in that it comprises a lysate of probiotic microorganisms in the form of a dry powder in the following ranges of percentages by weight relative to the total amount of probiotic microorganisms: - 15% to 20% of bacterial lysates of the genus Bacillus; - 15% to 35% of bacterial lysates of the genus Lactobacillus, - 1.5% to 8% of bacterial lysates of Streptococcus spp. - 45% to 60% of bacterial lysates of Saccharomyces spp., and - 1.5% to 8% of bacterial lysate of Bifidobacterium spp.

[0028] As used herein, the terms "postbiotic composition of the present invention," "orally administered composition," and "postbiotic" are synonymous and can be used interchangeably.

[0029] In the context of the present invention, bacterial lysates are understood as products obtained after a process of culturing and subsequent mechanical or chemical disruption of said bacterial cells to obtain a product containing bacterial fragments and all the components contained therein. Likewise, in this specification, these lysates are indicated as dried bacterial lysates, since, thanks to the method of their acquisition, they are subsequently subjected to drying techniques, and the dried bacterial lysates are in the form of a powder.

[0030] For the purposes of this document, degenerative eye diseases are understood as either 1) eye diseases that gradually destroy central and / or peripheral vision, leading to legal or total blindness within a few years, or 2) irreversible degenerative processes that result in a very poor quality of life for patients and make even the most basic tasks extremely difficult. The first type includes age-related macular degeneration, inherited retinal degenerative diseases, glaucoma, uveitis, and intraocular inflammatory diseases. The second type is primarily characterized by dry eye and ocular surface problems.

[0031] Similarly, in the context of the present invention, the term probiotic microorganisms defines live microorganisms, both bacteria and yeasts, that provide a health benefit when ingested.

[0032] In a preferred embodiment of the present invention, the composition of the present invention comprises a Bacillus bacterial lysate in an amount of between 15% and 20% by weight percentage. In a preferred embodiment, the amount of Bacillus bacterial lysate may be 15%, 16%, 17%, 18%, 19% or 20%.

[0033] In a preferred embodiment of the present invention, the composition of the present invention comprises a Lactobacillus lysate in an amount of 15% to 35%, preferably 18% to 30%, more preferably 20% to 28% by weight. In a preferred embodiment, the amount of Lactobacillus lysate can be 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30%.

[0034] In a preferred embodiment of the present invention, the composition of the present invention comprises a Streptococcus bacterial lysate in an amount of 1.5% to 8%, preferably 2% to 7%, and more preferably 3% to 6% by weight. In a preferred embodiment, the amount of Streptococcus bacterial lysate can be 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, or 3%. In another preferred embodiment, the amount of Streptococcus bacterial lysate can be 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, or 8%.

[0035] In a preferred embodiment of the present invention, the composition of the present invention comprises a weight percentage of Saccharomyces bacterial lysate of between 45% and 60%, preferably between 50% and 58%, more preferably between 54% and 56%. In a preferred embodiment, the amount of Saccharomyces bacterial lysate can be 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, or 60%.

[0036] In a preferred embodiment of the present invention, the composition of the present invention comprises a Bifidobacterium lysate in an amount of 1.5% to 8%, preferably 2% to 7%, and more preferably 3% to 6% by weight. In preferred embodiments, the amount of Bifidobacterium lysate can be 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, or 3%. In other preferred embodiments, the amount of Streptococcus lysate can be 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, or 8%.

[0037] In a preferred embodiment of the present invention, the bacterial lysate of the genus Bacillus is selected from the group consisting of Bacillus coagulans, Bacillus licheniformis, Bacillus mesentericus, Bacillus subtilis, Bacillus clausii, Bacillus paralicheniformis, and combinations thereof, preferably Bacillus coagulans, Bacillus licheniformis, Bacillus mesentericus, and Bacillus subtilis, or Bacillus licheniformis. The species is selected from the group consisting of Bacillus licheniformis and Bacillus subtilis.

[0038] In another embodiment, the bacterial lysate of the genus Lactobacillus is selected from the group consisting of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus plantarum, and the like. plantarum, Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus kefiri, and combinations thereof, preferably Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus reuteri, and Lactobacillus rhamnosus, or Lactobacillus acidophilus, Lactobacillus casei, and combinations thereof. casei, Lactobacillus fermentum, Lactobacillus reuteri, and Lactobacillus rhamnosusrhamnosus).

[0039] Furthermore, in other embodiments, the bacterial lysate of the genus Streptococcus is selected from the group consisting of Streptococcus thermophilus, Streptococcus salivarius, and the like. salivarius, and combinations thereof, preferably Streptococcus thermophilus.

[0040] In other specific embodiments of the present invention, the bacterial lysate of the genus Saccharomyces is a species selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces boulardii, and combinations thereof, preferably Saccharomyces cerevisiae.

[0041] In another preferred embodiment of the present invention, the bacterial lysate of the genus Bifidobacterium is selected from the group consisting of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis subsp. lactis, Bifidobacterium infantis, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis subsp. lactis, Bifidobacterium infantis, Bifidobacterium breve ... animalis, and combinations thereof, preferably a species selected from the group consisting of Bifidobacterium bifidum and Bifidobacterium lactis.

[0042] In a particular embodiment of the present invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - approximately 15% to 18% of bacterial lysates of the genus Bacillus; - approximately 3% to 7% of bacterial lysates of the genus Bifidobacterium, - approximately 17% to 30% of bacterial lysates of the genus Lactobacillus; - about 50% to 57% of yeast lysates of the genus Saccharomyces, - Approximately 2% to 5% of bacterial lysates of Streptococcus spp.

[0043] This composition is referred to as composition "A."

[0044] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - Approximately 15% to 17% of bacterial lysates of the genus Bacillus; - approximately 4.6% to 6% of bacterial lysates of the genus Bifidobacterium; - approximately 20% to 25% of bacterial lysates of the genus Lactobacillus; - about 54% to 56% of yeast lysates of the genus Saccharomyces; - Approximately 2.8% to 3.2% of bacterial lysates of Streptococcus spp.

[0045] This composition is referred to as composition "B."

[0046] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - Approximately 16% of bacterial lysates of Bacillus spp. - approximately 5% of bacterial lysates of the genus Bifidobacterium, - Approximately 21% of bacterial lysates of Lactobacillus spp. - about 55% of yeast lysates of Saccharomyces spp. - Approximately 3% of bacterial lysates of Streptococcus spp.

[0047] This composition is referred to as composition "C."

[0048] In a particular embodiment of the present invention, the composition comprises a lysate of a probiotic microorganism in the following amounts, expressed as a percentage by weight of the total lysate of the composition: - about 15% to 20% of a bacterial lysate of Bacillus licheniformis and Bacillus subtilis, and optionally Bacillus mesentericus and Bacillus coagulans, - Approximately 1.5% to 8% of Bifidobacterium bifidum and Bifidobacterium lactis lysates, - approximately 15% to 35% of a bacterial lysate of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus rhamnosus, and optionally Lactobacillus bulgaricus; - Approximately 45% to 60% of yeast lysate from Saccharomyces cerevisiae - Approximately 1.5% to 8% of Streptococcus thermophilus bacterial lysates.

[0049] This composition is referred to as composition "D."

[0050] In a particular embodiment of the present invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - approximately 15% to 18% of a bacterial lysate of Bacillus licheniformis and Bacillus subtilis, and optionally Bacillus mesentericus and Bacillus coagulans, - Bifidobacterium bifidum and Bifidobacterium lactis, approximately 3% to 7% of the bacterial population - approximately 17% to 30% of a bacterial lysate of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus rhamnosus, and optionally Lactobacillus bulgaricus; - Approximately 50% to 57% of yeast lysates from Saccharomyces cerevisiae - Approximately 2% to 5% of Streptococcus thermophilus bacterial lysates.

[0051] This composition is referred to as composition "E."

[0052] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - approximately 15% to 17% of a bacterial lysate of Bacillus licheniformis and Bacillus subtilis, and optionally Bacillus mesentericus and Bacillus coagulans, - approximately 4.6% to 6% of Bifidobacterium bifidum and Bifidobacterium lactis lysates, - approximately 20% to 25% of bacterial lysates of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus rhamnosus, and optionally Lactobacillus bulgaricus, - Approximately 54% to 56% of yeast lysates from Saccharomyces cerevisiae - Approximately 2.8% to 3.2% of Streptococcus thermophilus bacterial lysates.

[0053] This composition is designated as composition "F."

[0054] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - 1% to 20% Bacillus licheniformis and 1% to 12% Bacillus subtilis, and optionally 1% to 10% Bacillus mesentericus and 0.1% to 10% Bacillus coagulans, - Bifidobacterium lactis: 0.3% to 10%, and Bifidobacterium bifidum: 0.1% to 8%, - 0.5% to 22% Lactobacillus acidophilus, 0.1% to 12% Lactobacillus casei, 0.1% to 12% Lactobacillus fermentum, 0.1% to 6% Lactobacillus reuteri, 0.1% to 6% Lactobacillus rhamnosus, and optionally 0.2% to 10% Lactobacillus bulgaricus, - Saccharomyces cerevisiae: 35% to 75% - Streptococcus thermophilus: 0.1% to 8%.

[0055] This composition is referred to as composition "G."

[0056] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - Bacillus licheniformis 2% to 15% and Bacillus subtilis 2% to 10%, and optionally Bacillus mesentericus 2% to 8% and Bacillus coagulans 0.2% to 8%, - Bifidobacterium lactis: 0.5% to 8%, Bifidobacterium bifidum: 0.5% to 7%, - 1% to 20% Lactobacillus acidophilus, 0.5% to 10% Lactobacillus casei, 0.3% to 10% Lactobacillus fermentum, 0.5% to 5% Lactobacillus reuteri, 0.3% to 5% Lactobacillus rhamnosus, and optionally 0.5% to 8% Lactobacillus bulgaricus; - Saccharomyces cerevisiae: 40% to 70% - Streptococcus thermophilus: 0.3% to 6%.

[0057] This composition is referred to as composition "H."

[0058] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - Bacillus licheniformis 3% to 11% and Bacillus subtilis 3% to 9%, and optionally Bacillus mesentericus 3% to 7% and Bacillus coagulans 0.3% to 7%, - Bifidobacterium lactis: 1% to 7%, Bifidobacterium bifidum: 0.8% to 6%, - 2% to 18% Lactobacillus acidophilus, 0.5% to 9% Lactobacillus casei, 0.5% to 9% Lactobacillus fermentum, 0.7% to 4.5% Lactobacillus reuteri, 0.5% to 4.5% Lactobacillus rhamnosus, and optionally 0.7% to 7% Lactobacillus bulgaricus, - Saccharomyces cerevisiae: 43% to 68% - Streptococcus thermophilus: 0.5% to 5.5%.

[0059] This composition is referred to as composition "I."

[0060] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - 4% to 11% Bacillus licheniformis and 4% to 8% Bacillus subtilis, and optionally 3% to 6% Bacillus mesentericus and 0.4% to 6% Bacillus coagulans, - Bifidobacterium lactis: 1.5% to 6%, Bifidobacterium bifidum: 0.8% to 5%, - 2.5% to 15% Lactobacillus acidophilus, 0.5% to 8% Lactobacillus casei, 1% to 8% Lactobacillus fermentum, 0.9% to 4% Lactobacillus reuteri, 0.7% to 4% Lactobacillus rhamnosus, and optionally 1% to 5% Lactobacillus bulgaricus, - Saccharomyces cerevisiae: 45% to 65%, - Streptococcus thermophilus: 1% to 5%.

[0061] This composition is referred to as composition "J."

[0062] In another particular embodiment of the invention, the composition comprises the following amounts of lysate of a probiotic microorganism, expressed as a percentage by weight relative to the total lysate of the composition: - Bacillus licheniformis 4.5% to 10.5% and Bacillus subtilis 4.5% to 8%, and optionally Bacillus mesentericus 3% to 6% and Bacillus coagulans 0.4% to 3%, - Bifidobacterium lactis: 2% to 5.5%, Bifidobacterium bifidum: 0.8% to 3%, - 3% to 12% Lactobacillus acidophilus, 0.6% to 7% Lactobacillus casei, 2% to 7% Lactobacillus fermentum, 1% to 3% Lactobacillus reuteri, 1% to 4% Lactobacillus rhamnosus, and optionally 2% to 5% Lactobacillus bulgaricus; - Saccharomyces cerevisiae: 50% to 60%, - Streptococcus thermophilus: 2% to 4%.

[0063] This composition is referred to as composition "K."

[0064] In certain embodiments of the present invention, the composition for use comprises a lysate of a probiotic microorganism in the form of a dry powder in an amount as a percentage by weight of the total as disclosed in compositions "A", "B", "C", "D", "E", "F", "G", "H", "I", "J" or "K".

[0065] In other specific embodiments of the present invention, the compositions of the present invention may comprise another additional ingredient or additive selected from the group consisting of vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B9, vitamin B12, zeaxanthin, hydroxytyrosol, omega-3 fatty acid (DHA), glutathione, copper, selenium, and combinations thereof.

[0066] Below is given the amount that each of the additional components or additives that may optionally comprise the composition according to the first aspect of the present invention may be included.

[0067] Table 1. List of additional ingredients that may comprise the composition subject matter of the present invention, and amounts in particularly preferred and most preferred embodiments.

[0068] [Table 1]

[0069] Additional ingredients included in the composition can be products to cover common vitamin deficiencies that usually occur in people from the age of 50, such as vitamins B1 and B3, which help increase the normal energy performance of the metabolism and maintain the correct functioning of the defense system.

[0070] Antioxidant compounds such as hydroxytyrosol, a polyphenol found in olive fruit that protects against oxidative stress, selenium, a trace element necessary for antioxidant defense, and glutathione, which is part of the body's endogenous system along with the enzymes superoxide dismutase and glutathione peroxidase, further contribute to protection against oxidative stress.

[0071] Contains minerals necessary for bone maintenance, magnesium necessary for muscle function, soluble dietary fiber, and omega-3 fatty acids that improve memory.

[0072] Finally, another component of note is copper, which contributes to the maintenance of normal connective tissue.

[0073] In the context of the present invention, the methods by which the compositions of the present invention can be obtained are also described.

[0074] The postbiotic consists of lysates of probiotic microorganisms including Bacillus, Lactobacillus, Streptococcus, Saccharomyces, and Bifidobacterium.

[0075] Due to the fact that the composition object of the present invention is in powder form, the composition according to the first aspect of the present invention can itself be presented in a sealed pouch as is customary in the food and pharmaceutical industries.

[0076] Another form of food supplement according to the present invention is in the form of a capsule, such as a conventional gelatin capsule, containing the composition in powder form.

[0077] In a preferred embodiment of the present invention, the composition may have a relative abundance of microbial protein from a Bacillus bacterial lysate of between 4% and 6%, preferably between 4.14% and 5.61%, and more preferably between 4.21% and 5.27% by weight of protein from a Bacillus bacterial lysate. In a preferred embodiment, the relative abundance of protein from a Bacillus bacterial lysate may be 4%, 4.20%, 4.40%, 4.60%, 4.80%, 5%, 5.20%, 5.40%, 5.60%, 5.80%, or 6%.

[0078] In a preferred embodiment of the present invention, from a bacterial lysate of the genus Bacillus, the identified protein is selected from the group consisting of Q65HF3, P04831, P04832, and combinations thereof.

[0079] In a preferred embodiment of the present invention, the composition may comprise a relative abundance of proteins from a Bifidobacterium bacterial lysate relative to total microbial proteins by weight of between 0.02% and 0.10%, preferably between 0.07% and 0.09%, hi a preferred embodiment, the relative abundance of proteins from a Bifidobacterium bacterial lysate may be 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.10%.

[0080] In a preferred embodiment of the present invention, the proteins identified from bacterial lysates of the genus Bifidobacterium are selected from the group consisting of B8DTX9, B8DSQ4, and combinations thereof.

[0081] In a preferred embodiment of the present invention, the composition may comprise a protein from a bacterial lysate of the genus Lactobacillus, the relative abundance of which is at least 3% to 8.50%, preferably 3.45% to 8.06%, more preferably 4.84% to 5.45% by weight of the total microbial protein. In a preferred embodiment, the relative abundance of the protein from a bacterial lysate of the genus Lactobacillus may be 3%, 3.50%, 4%, 4.50%, 5%, 5.50%, 6%, 6.50%, 7%, 7.50%, 8% or 8.50%.

[0082] In a preferred embodiment of the present invention, the proteins identified from bacterial lysates of the Lactobacillus genus are selected from the group consisting of A0A0E2BRTT9, D8IHB6, Q5FKM6, and combinations thereof.

[0083] In a preferred embodiment of the present invention, the composition may comprise between 70% and 99.00%, preferably between 73.85% and 98.5%, and more preferably between 85% and 90.30% by weight of protein from a bacterial lysate of Saccharomyces, as a percentage of total microbial protein abundance. In a preferred embodiment, the relative abundance of protein from a bacterial lysate of Saccharomyces may be 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, or 89%. In other preferred embodiments, the relative abundance of the protein from a bacterial lysate of Saccharomyces genus may be 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5% or 99%.

[0084] In a preferred embodiment of the present invention, the proteins identified from bacterial lysates of Saccharomyces are selected from the group consisting of A0A0L8VPY0, P00359, A0A0L8VN66, A0A0L8VP44, and combinations thereof.

[0085] In a preferred embodiment of the present invention, the composition may comprise a protein from a bacterial lysate of the genus Streptococcus present in an amount of 0.15% to 0.90%, preferably 0.17% to 89%, and more preferably 0.33% to 66% by weight relative to the total amount of microbial protein. In a preferred embodiment, the relative abundance of the protein from a bacterial lysate of the genus Streptococcus may be 0.15%, 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, 0.55%, 0.60%, 0.65%, 0.70%, 0.75%, 0.80%, 0.85%, or 0.90%.

[0086] In a preferred embodiment of the present invention, the protein identified from a bacterial lysate of the genus Streptococcus is selected from the group consisting of Q5M561, Q5M518, and combinations thereof.

[0087] To obtain the compositions of the present invention, a culture of a genera described herein is first prepared containing a certain amount of colony forming units (CFU). In the context of the present invention, colony forming units is a microbiological term that indicates the amount of live microorganisms present in a culture medium.

[0088] In a particular embodiment of the present invention, the composition starts with a bacterial population of the genus Bacillus comprising 10.56% to 14.08% CFU, most preferably 11.26% to 12.75% CFU of the total CFU of the composition.

[0089] In a particular embodiment of the present invention, the composition starts with a Lactobacillus population comprising between 29.40% and 68.61% CFU, most preferably between 41.17% and 53.36% CFU of the total CFU of the composition.

[0090] In a particular embodiment of the present invention, the starting population of Streptococcus comprises between 5.83% and 31.07% CFU, most preferably between 11.65% and 18.15% CFU of the total CFU of the composition.

[0091] In a particular embodiment of the present invention, the starting population of Saccharomyces comprises between 17.48% and 23.30% CFU, most preferably between 21.36% and 22.45% CFU of the total CFU of the composition.

[0092] In a particular embodiment of the present invention, the composition starts with a Bifidobacterium population comprising between 4.37% and 23.30% CFU, most preferably between 14.56% and 17.35% CFU of the total CFU of the composition.

[0093] These probiotic bacteria are listed in the Spanish Collection of Types of Bacteria, which are listed for each bacterial species described in this book. The cells are cultured under standard conditions as described in the culture protocol published by the Center for Epithelial Cell Cultures (CECT).

[0094] Once cultured, these microorganisms undergo a lysis process. The process for obtaining the lysate involves a combination of non-mechanical and mechanical methods. First, the microbial cells undergo a heat treatment. Each batch of live cell culture undergoes a sterilization cycle in an autoclave at 121°C for 20 to 30 minutes. This temperature denatures, coagulates, and inactivates proteins. It also causes membrane damage, ribosome aggregation, DNA strand breaks, and enzyme inactivation. Once cooled, the probiotic batch undergoes cell disruption by sonication for 15 to 20 minutes at 450 to 550 W, 38 to 43% amplitude, and 8 to 15 seconds of pause time (Qsonica, Q500). This results in disruption of intermolecular interactions and DNA fragmentation. The final solution is freeze-dried and then milled to obtain the probiotic microbial lysate in powder form. The powder is stored in a cool, protected environment.

[0095] In a second aspect, the present invention relates to the use of the composition of the present invention as a food supplement for preventing and / or delaying ocular degenerative diseases. In a preferred embodiment, the ocular degenerative diseases are selected from the group consisting of age-related macular degeneration, inherited retinal degenerative diseases, glaucoma, uveitis or intraocular inflammatory diseases, dry eye, ocular surface changes, and combinations thereof.

[0096] In a third aspect, the present invention relates to a composition as defined herein for use as a medicament.

[0097] In a fourth aspect, the present invention relates to a composition as defined herein for use in the treatment of a degenerative disease of the eye.

[0098] The degenerative eye disease is one or more of the following: age-related macular degeneration (AMD), retinitis pigmentosa, glaucoma, corneal dystrophy, cataract, Stargardt disease, Leber's disease, keratoconus, Best's disease, and optic atrophy.

[0099] In certain embodiments, the degenerative disease may be in one or both eyes.

[0100] In a preferred embodiment, the degenerative eye disease is age-related macular degeneration (AMD), and unless otherwise specified, AMD can be both wet AMD and dry AMD at any stage of progression, such as early intermediate or advanced, with drusen in any state, such as soft drusen, hard drusen, mixed drusen, and reticular drusen.

[0101] In a fifth aspect, the present invention relates to a pharmaceutical composition comprising an effective pharmaceutical amount of a composition according to the first aspect of the invention, and a pharmaceutically acceptable excipient.

[0102] In the context of the present invention, the expression "pharmaceutical composition" refers to a formulation adapted to deliver a predetermined amount of one or more useful therapeutic agents to a cell, group of cells, organ or tissue.

[0103] The term "effective pharmaceutical amount" as used herein is understood as an amount that can provide a therapeutic effect and can be determined by commonly used means by those skilled in the art.

[0104] Also, in the context of the present invention, "pharmaceutically acceptable excipient" means a therapeutically inactive substance used to incorporate an active ingredient, which is acceptable to the patient from a pharmacological / toxicological point of view and acceptable to the pharmaceutical chemist who produces it from a physical / chemical point of view in terms of composition, formulation, stability, patient acceptability and bioavailability.

[0105] In the context of the present invention, food supplements are any substances intended to supplement the normal diet and consisting of concentrated sources of nutrients or other substances having a nutritional or physiological effect, in simple or combined form, administered in small unit doses, i.e. in capsules, tablets, pastilles and other similar forms, powder sachets, liquid ampoules, in liquid form, as defined in Directive 2002 / 46 / EC of the European Parliament. , a food product sold in dropper bottles and other similar forms.

[0106] In certain embodiments of the present invention, the composition for treating macular degeneration comprises that the dosage is orally administered to patient at least twice a day.And, in order to be effective in treatment and achieve desired effect, this dosage should be administered to patient at least twice a day, and at most 6 times a day.Preferably, the dosage is administered to patient 3 times, 4 times or 5 times a day, and most preferably 3 times a day.

[0107] In the context of the present invention, a dose is defined as the amount of a pharmaceutical agent containing the correct active ingredient so as to be efficient, effective and safe for the patient and to solve the indicated health problem.

[0108] Therefore, in the context of the present invention, a dose is determined to contain an amount of the composition of the present invention between 100 mg and 400 mg. In a preferred embodiment, a dose may contain 150 mg to 300 mg of the composition. In certain embodiments of the present invention, a dose may contain 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg of the composition of the present invention.

[0109] In certain embodiments in which the composition is provided in a watertight sachet, the composition can be administered to a patient dissolved or suspended in a liquid, preferably an aqueous liquid, more preferably a beverage such as fruit juice, milk, water, etc. It can also be mixed with foods such as yogurt, liquid yogurt, soup, puree, cream, or porridge. These foods must be at an optimal temperature for consumption and should never be heated after the composition of the present invention has been added.

[0110] In certain embodiments of the present invention, the food supplement described herein is orally administered to patients with macular degeneration at a single dose at least twice a day.And, in order to be effective in treatment and achieve desired effect, this dose needs to be administered to patients at least twice a day, and at most 6 times a day.Preferably, patient is administered 3 times, 4 times or 5 times a day, and most preferably 3 times a day.

[0111] In certain embodiments where the composition is provided in a watertight sachet, the food supplement can be administered to a patient by dissolving or suspending it in a liquid, preferably an aqueous liquid, more preferably a beverage such as fruit juice, milk, or water. It can also be mixed with foods such as yogurt, liquid yogurt, soup, puree, cream, or porridge. These foods must be at an optimal temperature for consumption and must never be heated after the composition of the present invention is added.

[0112] The subject of the composition of the present invention is an extract produced through the culture and lysis process of probiotic microorganisms, and contains a mixture of components such as metabolites, proteins, DNA fragments, peptidoglycans, etc. When administered at an appropriate dose, this extract has the effect of altering the host's microbiota through multiple mechanisms, inducing immune system activation, and naturally reversing dysbiosis.

[0113] The technical effect obtained from the composition of the present invention is to directly affect the microbiota of patients with macular degeneration, regulating and improving said microbiota. Thanks to this restorative and regulating effect, the inventors have surprisingly confirmed that by intervening in the microbiota of patients with macular degeneration, it is possible to significantly improve the condition of this disease and slow down its progression.

[0114] In this way, the inventors have been able to confirm that oral administration of the compositions of the invention exerts an effect on the modulation and reprogramming of the microbiota present in the intestinal tract of a patient that is not just a local effect, since the whole organism benefits from this modulation of the microbiota, and therefore is a systemic effect.

[0115] Thus, the main advantages resulting from the composition subject of the present invention are: - supplementing and / or complementing the physiological function of the patient's microbiome; - reversal of changes that result in dysbiosis and have clinical consequences in macular diseases of the human eye, particularly age-related macular degeneration (AMD); - due to the fact that the product is administered orally, the composition and its remarkable effects are presented as a very advantageous alternative compared to the above-mentioned treatments currently used in medicine, such as injections of drugs into the eye, which, in addition to being painful, require the intervention of a specialist; - The composition contains lysates of microorganisms, i.e. it does not present living organisms, and therefore presents itself as a safe alternative, since it avoids the possible dangers posed by colonizing organisms and, moreover, does not present the toxicity that conventional drugs may present.

[0116] Furthermore, the present invention also refers to the following clauses:

[0117] 1. An orally administered composition for preventing and treating degenerative eye diseases, comprising a lysate of probiotic microorganisms in the form of a dry powder, the amount of which, as a percentage by weight of the total, is comprised between: - 15% to 20% of bacterial lysates of the genus Bacillus; - 15% to 35% of bacterial lysates of the genus Lactobacillus, - 1.5% to 8% of bacterial lysates of Streptococcus spp. - 45% to 60% of bacterial lysates of Saccharomyces spp., and - 1.5% to 8% of bacterial lysate of Bifidobacterium spp.

[0118] 2. The composition of clause 1, wherein the bacterial lysate of the genus Bacillus is a species selected from the group consisting of Bacillus coagulans, Bacillus licheniformis, Bacillus mesentericus, Bacillus subtilis, Bacillus clausiai, Bacillus paralicheniformis, and combinations thereof.

[0119] 3. The composition of clause 1 or 2, wherein the bacterial lysate of the genus Bifidobacterium is a species selected from the group consisting of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis subsp. lactis, Bifidobacterium infantis, Bifidobacterium animalis, and combinations thereof.

[0120] 4. The composition of any of clauses 1 to 3, wherein the Lactobacillus bacterial lysate is a species selected from the group consisting of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus kefiri, and combinations thereof.

[0121] 5. The composition of any of clauses 1 to 4, wherein the bacterial lysate of the genus Saccharomyces is a species selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces boulardii, and combinations thereof.

[0122] 6. The composition of any of clauses 1 to 5, wherein the bacterial lysate of the genus Streptococcus is a species selected from the group consisting of Streptococcus salivarius, Streptococcus thermophilus, and combinations thereof.

[0123] 7. The composition of any of clauses 1 to 6, comprising at least one additional ingredient selected from the group consisting of vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B9, vitamin B12, vitamin C, vitamin E, zeaxanthin, hydroxytyrosol, omega-3 fatty acid (DHA), glutathione, copper, selenium, zinc, lutein, and combinations thereof.

[0124] 8. The composition according to any of clauses 1 to 7, characterized in that it contains, as weight percentages, the following relative amounts present: - 4% to 6% of the proteins from bacterial lysates of Bacillus spp. - 0.02% to 0.10% of proteins from a bacterial lysate of the genus Bifidobacterium; - 3% to 8.50% of proteins from bacterial lysates of Lactobacillus genus, - 70% to 99.00% of the proteins from bacterial lysates of Saccharomyces spp. - 0.15% to 0.90% of the protein from bacterial lysates of Streptococcus spp.

[0125] 9. Use of a composition according to any of clauses 1 to 8 as a food supplement for the prevention and / or delay of degenerative eye diseases.

[0126] 10. Use of the composition according to clause 9, wherein the composition is administered orally in an amount between 100 mg and 400 mg, one to six times daily.

[0127] 11. Use of a composition according to clauses 9 or 10, wherein the composition is presented in powder form in a watertight sachet or enclosed in a gelatin capsule.

[0128] 12. A composition according to any one of clauses 1 to 8 for use as a medicament.

[0129] 13. The composition according to clause 12 for use in the prevention and treatment of degenerative eye diseases.

[0130] 14. The composition of clause 12 or 13, wherein the composition is administered orally in an amount between 100 mg and 400 mg of the composition 1 to 6 times daily.

[0131] 15. A pharmaceutical composition comprising an effective pharmaceutical amount of the composition according to any one of clauses 1 to 8 and a pharmaceutically acceptable excipient.

[0132] The terms "comprising," "consisting essentially of," and "consisting of" may be interchanged with either of the other two terms. The terms "a" or "an" can refer to one or more of the elements it modifies, unless the context clearly indicates that any element or elements are being described (e.g., "a reagent" can mean one or more reagents). As used herein, the term "about" refers to a value within 10% of the underlying parameter (i.e., plus or minus 10%; e.g., a weight of "about 100 grams" can include weights between 90 grams and 110 grams). The use of the word "about" at the beginning of a list of values ​​modifies each value (e.g., "about 1, 2, and 3" refers to "about 1, about 2, and about 3"). When a list of values ​​is stated, the list includes all intermediate values ​​and all fractional values ​​thereof (e.g., the list of values ​​"80%, 85%, or 90%" includes the intermediate value 86% and the fractional value 86.4%). When a list of values ​​is followed by the term "or more," the term "or more" applies to each of the listed values ​​(e.g., "80%, 90%, 95% or more" or "80%, 90%, 95% or more" or "80%, 90%, or 95% or more" means "80% or more, 90% or more, or 95% or more"). When a list of values ​​is given, the list includes all ranges between any two of the listed values ​​(e.g., the list "80%, 90% or 95%" includes the ranges "80% to 90%, "80% to 95%, and "90% to 95%"). Specific embodiments of the present technology are described in the examples below.

[0133] Preferred Embodiments of the Invention For the purpose of providing a better understanding of the present invention and in accordance with its practical realization, preferred embodiments of the present invention have been described as an integral part of this specification.

[0134] Example 1 - Preparation of a lysate composition of probiotic microorganisms

[0135] The process involves obtaining a mixture of dried microbial lysates of different strains of Bacillus, Lactobacillus, Bifidobacterium, Streptococcus, and Saccharomyces. To achieve this, they are grown by fermentation, concentrated, and digested, and finally the cellular components are dried. The resulting postbiotic formulation is a lysate mixture that can be taken with food ingredients in the form of a capsule or powder supplement.

[0136] For growth, prepare a sufficient volume and amount of inoculum in the culture medium flask for each microorganism to inoculate the fermenter with the cultured inoculum. To do this, immerse the microorganisms stored in vials with 20% glycerol at -80°C in a 30°C water bath to quickly and completely thaw each microorganism.

[0137] Once the inoculum of each microorganism was obtained, it was inoculated separately directly into the fermenter at an inoculation rate of 3% for Bacillus, 5% for Bifidobacterium, 5% for Lactobacillus, 5% for Streptococcus, and 5% for Saccharomyces.

[0138] After inoculating the fermentor with the corresponding microbial inoculum, the fermentation step for each microbial species is carried out to generate microbial biomass, and growth conditions for each species are established.

[0139] Bacillus was cultured in Nutrient Broth (NB) at 30°C for 15 hours at 250 rpm. The medium was glucose HO 25g / L, yeast extract 5g / L, KHPO4 The fermentation medium consists of 2.5 g / L ammonium hydroxide, 5 g / L MgSO₄·7H₂O, and 1 g / L Tween 80. The incubation is carried out at 30°C ± 1°C throughout the entire process, with a pH of 6.8 ± 0.1 maintained by the automatic aseptic addition of 25% NH₄OH or 35% H₃PO₄. Dissolved oxygen is maintained above 40% by agitation (150 to 250 rpm) at 0.5 atmospheres and aeration (1 to 1.5 vvm). After approximately 20 to 24 hours, base consumption (NH₄OH) is stopped, the temperature is lowered to 4 to 8°C to stop cellular metabolism, and the broth is harvested.

[0140] For Streptococcus, the inoculum is grown in tryptic soy broth (TSB) at 37°C with 150 rpm agitation for 24 hours. The fermentation medium consists of 25 g / L glucose, 8 g / L milk powder, 5 g / L yeast extract, 2 g / L K2HPO4, 2 g / L KH2PO4, 0.5 g / L MgSO4´7H2O, and 0.5 g / L (NH4)SO4. The growth temperature is 37°C ± 1°C, and the pH is 0.05. The pH is maintained at 6.8±0.1 throughout the process by the automated aseptic addition of 25% H₃PO₄35%. Agitation is at 150 rpm. After fermentation is complete, approximately 20 to 24 hours later, the temperature is lowered to 4 to 8°C to stop cell metabolism and the broth is harvested.

[0141] Bifidobacteria and Lactobacillus were grown in De Man, Rogosa, and Sharp Broth (MRS Broth) at 37°C for 24 hours. Fermentations were performed at 37°C ± 1°C, pH 6.2 ± 0.1 and 6.4 ± 0.1, with the aseptic automatic addition of 25% NH4OH or 35% H3PO4, and at 50 rpm (the minimum required to ensure accurate homogenization of the medium components). Yeast extract 5g / L, K2HPO4 2g / L, and casein peptone were added. 10g / l, milk powder 8g / l, sodium acetate 5g / l, diammonium citrate 2g / l, Mn SO4- H2O The fermentation conditions are: 0.05g / l, MgSO₄ 7H₂O 0.2g / l, Tween 80 1g / l. Nitrogen is initially bubbled in until the dissolved oxygen concentration reaches 0%, and this is repeated as necessary to maintain this level throughout the process. At the end of the fermentation (approximately 20 to 24 hours), the temperature is lowered to 4 to 8°C to stop cell metabolism and the broth is harvested.

[0142] S. cerevisiae was grown in YPD for 24 hours. The fermenter temperature was maintained at 30°C, pH 5 at 150 rpm, and 1.5 vvm. Fermentation was stopped in the fermenter, and the temperature was lowered to 4-8°C.

[0143] The biomass obtained from the fermentation of each microorganism is recovered by centrifugation or microfiltration to obtain wet recovered biomass, which is concentrated as many times as necessary to obtain 1.00E+09 to 1.00E+11 cfu / ml in the concentrated biomass of each microorganism.

[0144] The cells of the wet harvested biomass are then ruptured or lysed by heat treatment and ultrasonic treatment to obtain the respective bacterial lysates, which are then dried separately in a freeze dryer or sprayer, and the dried bacterial lysates are mixed in a mixer to obtain the adjusted composition.

[0145] Each batch of live cell culture is autoclaved for 20 minutes at 121°C. It is then sonicated for 20 minutes at 500W, 40% amplitude, and a 10-second pause to obtain the lysed probiotic cell mass (Qsonica, Q500). The final solution is freeze-dried and pulverized to obtain the powdered probiotic microbial lysate. The powder is stored in a cool, protected environment.

[0146] For lysate analysis, 0.5g of powder was taken from each lot and dissolved in 10ml of distilled water. A 1:8 dilution was prepared from this suspension (100ul of sample and 700ul of water) and centrifuged at 5000rpm for 5 minutes. The supernatant was collected and 1.5ul of sample was placed in a cuvette. Distilled water was used as a blank for the Nanodrop Tecan Spark assay. Measure the absorbance at 260 nm, 230 nm, and 280 nm at 10 M. Perform the measurement at least twice, and usually repeat the measurement four or more times.

[0147] The maximum absorption peak for the quantitative determination of DNA, a nucleic acid, occurs at a wavelength of 260 nm, so the absorption at that wavelength is proportional to the DNA concentration. (ng / ul DNA: A260nm x dilution factor x 50 (conversion factor).

[0148] The amount of DNA contained in each lysate is as follows:

[0149] Table 2. DNA concentrations of dried lysates of each bacterial strain prepared according to the procedure.

[0150] [Table 2]

[0151] The following dried bacterial lysates were used in the following proportions to prepare samples of the compositions of the present invention:

[0152] Table 3. Composition of Sample 1 of the composition of the present invention. The weight percentage of each genus described herein is given.

[0153] [Table 3]

[0154] In a further preferred embodiment, to determine the effectiveness of the composition of the present invention in AMD patients, a study was conducted in patients with this disease, in which the dosage of the composition of the present invention was 400 mg three times daily.

[0155] In particular, a composition of the invention consisting of dried bacterial lysates of the following species was administered: amounts given as percentages by weight:

[0156] Table 4. Composition of Sample 1 of the composition of the present invention. The weight percentage of each bacterial species that makes up the dried bacterial lysate described herein is shown.

[0157] [Table 4]

[0158] Capsule 1. Example of AMD treatment capsules Gelatin capsules were prepared containing the following ingredients:

[0159] Table 5. Examples of capsule compositions

[0160] [Table 5]

[0161] Patients took one to three capsules daily.

[0162] Capsule 2. Example of a capsule for treating AMD Gelatin capsules were prepared containing the following ingredients:

[0163] Table 6. Examples of capsule compositions

[0164] [Table 6]

[0165] Patients took one to three capsules daily.

[0166] Capsule 3 - Example of a capsule for treating AMD Gelatin capsules were prepared containing the following ingredients:

[0167] Table 7. Examples of capsule compositions

[0168] [Table 7]

[0169] Patients took one to three capsules daily.

[0170] Example 2 - Analysis of the composition prepared in Example 1

[0171] 2.1. Sample preparation and protein digestion 400 mg of the composition of Example 1 (hereinafter referred to as the sample) was added to 8.4 M urea (USB The cells were dissolved in chaotropic buffer containing 2.4 M thiourea (Sigma-Aldrich), 5% CHAPS (Sigma-Aldrich), 5 mM TCEP (Sigma-Aldrich), and a protease inhibitor cocktail (Sigma-Aldrich), and incubated on ice for 15 minutes. Homogenization was performed by sonication in a Branson 2510 bath (Marshall Scientific, New Hampshire, USA) for 5 minutes. The homogenate was centrifuged at 20,000 × g for 10 minutes at 4°C, and the supernatant containing the solubilized protein was used for further analysis. 40 μg of protein was precipitated using the methanol / chloroform method and dissolved in 40 μg of UTT buffer (7 M urea, 2 M thiourea, 10 mM The samples were resuspended in a multi-chaotropic sample solution of TEAB (Sigma-Aldrich).

[0172] The resuspended sample was reduced with 2 μL of 50 mM TCEP at 37°C for 60 min, followed by addition of 1 μL of MMTS reaction mixture (SCIEX, Cysteine ​​blocker (Foster City, CA, USA) was added and incubated for 10 minutes at room temperature. To reduce the urea concentration, the sample was diluted to 140 μl with TEAB (mM). Finally, digestion was initiated by adding 2 μg of MS-grade Pierce trypsin (Thermo-Fisher Scientific, Inc., Waltham, MA, USA) to each fraction at a 1:20 (w / w) ratio and incubating overnight at 37°C in a shaker. The digested samples were evaporated to dryness in a vacuum concentrator.

[0173] 2.2. Liquid Chromatography and Mass Spectrometry (LC-MS) The digested samples were cleaned and desalted using Stage-Tips with Empore 3M C18 (Sigma-Aldrich) discs. A 1 μg aliquot of the resulting peptides was analyzed using an Eksigent Technologies nanoLC Ultra 1D plus nano liquid chromatography system (SCIEX, Foster City, CA, USA) coupled to a Triple TOF 5600 (SCIEX) high-speed mass spectrometer equipped with a Nanspray III source. LC ESI-MS / MS(Liquid Chromatography Electrospray Ionization Tandem Mass The analytical column was an Acquity UPLC M-Class Peptide BEH C18 reversed-phase silica-based column (Waters Corporation, Milford, MA, USA). The trap column was a C18 Acclaim PepMap™ 100 (Thermo-Fisher A 100 μm × 2 cm, 5 μm particle size, 100 Å pore size column (Scientific Inc.), 100 μm × 2 cm, 5 μm particle size, 100 Å pore size, was connected in-line with the analytical column. A charge pump delivered 0.1% formic acid in water at 2 μl / min. A flow rate of 250 nL was applied by a nanopump operated under gradient elution conditions. Peptides were separated using a 250-minute gradient ranging from 2% to 90% B phase (mobile phase A: 2% acetonitrile (Scharlab, SL, Spain), 0.1% formic acid (Sigma-Aldrich); mobile phase B: 100% acetonitrile, 0.1% formic acid). The injection volume was 5 μl. Data were collected using an ionspray float voltage of 2300 V, curtain gas of 35, interface heater temperature of 150, source gas of 125, and degrouper voltage of 150. V was used. For IDA (Intelligent Data Analysis) parameters, the values ​​ranged from 350 to 1250. A 0.25 s MS scan in the mass range of Fa was followed by 35 100 ms MS / MS scans in the mass range of 100 to 1800. Conversion criteria were established for ions with mass / charge ratio (m / z) greater than 350, m / z less than 1250, charge states between 2 and 5, and an abundance threshold greater than 90 counts per second (cps). The target ions were rejected for 15 seconds.

[0174] 2.3. Proteome data analysis and sequence searching The obtained spectroscopic data were processed using PeakView v2.2 (SCIEX) software and exported to mgf files, which were then analyzed using Mascot Server v2.5.1 (Matrix) against a protein database containing protein sequences of microorganisms of the genera Bacillus, Lactobacillus, Bifidobacterium, Streptococcus, and Saccharomyces from the Uniprot / Swissprot knowledge base (https: / / www.uniprot.org / statistics / Swiss-Prot-update:20170412, 2,542,118 protein sequences). The LC-MS analysis was performed using the Proteomics Facility at the National Biotechnology Centre of the High Council for Scientific Research, London, United Kingdom. Data were obtained for each protein from the total number of matching peptide spectra for a given protein (referred to as peptide-pair-spectrum matching (PSM) and exponentially corrected protein abundance index (emPAI)). By matching peptides in the resulting database, PSM and emPAI can be used as relative quantitative results for proteins in complex mixtures based on protein coverage. LC-MS analysis was performed at the Proteomics Facility of the National Biotechnology Centre of the High Council for Scientific Research, part of ProteoRed.

[0175] 2.4. Protein abundance and function in samples The samples were thoroughly analyzed for expressed proteins, resulting in the identification of 937 proteins. Spectral counts were obtained for each of the 937 identified proteins to obtain an indication of their abundance. Normalized emPAI values ​​(nemPAI%) were calculated based on the emPAI values ​​by dividing each individual value by the sum of all emPAI values ​​and multiplying each value by 100% to determine the relative abundance of each protein relative to the total protein.

[0176] Furthermore, to analyze the relative abundance of specific proteins in specific metabolic processes as defined by the Kyoto Encyclopedia of Genes and Genomes (KEGG) and COG ( http: / / eggnogdb.embl.de / # / app / home ), we performed functional analysis of proteins in the samples by assigning k-numbers using the KEGG internal annotation tool for KEGG orthology (KO) and searching for clusters of orthology groups (COG).

[0177] The distribution of nemPAI values ​​is shown in Figure 1. As can be seen, the distribution obtained has an exponential shape, indicating the presence of a great diversity of proteins in the sample.

[0178] The microbial origin of the samples was identified by a comparative search of the mass spectrometry data of the samples against proteins from the Uniprot / Swissprot database of microorganisms of the genera Bacillus, Lactobacillus, Bifidobacterium, Streptococcus, and Saccharomyces. The bacterial origin and contribution to the sample proteome are summarized in the table below.

[0179] Table 8. Mass spectrometry data of samples for proteins of microorganisms of the genera Bacillus, Lactobacillus, Bifidobacterium, Streptococcus, and Saccharomyces from the Uniprot / Swissprot database, where the bacterial origin and contribution to the sample proteome is determined.

[0180] [Table 8]

[0181] As can be seen, microorganisms of the genus Saccharomyces are the largest contributors, both in terms of the total number of proteins and the expression levels of the proteins present in the samples. Of the 937 proteins identified, 10 proteins with a relative abundance of 1% or more belong to this genus. These proteins and their nemPAI values ​​are listed in the table below.

[0182] Table 9. 937 proteins identified in the sample, 10 with a relative abundance of 1%

[0183] [Table 9]

[0184] Functional analysis based on KEGG orthology (KO) confirmed the protein functions in various metabolic pathways defined by KEGG and COG. Of the 937 proteins, 643 proteins were classified into 352 functional categories. Among the more abundant proteins, notable functions were those represented by category K01689 (enolase [EC: 4.2.1.11]), which corresponds to the E7LV64 protein. This enolase catalyzes the reversible conversion of 2-phosphoglycerate to phosphoenolpyruvate and is essential for carbohydrate degradation via glycolysis. Notable functions in the protein profile were K03530 (histone-like binding protein), K13953 (alcohol dehydrogenase), and K03671 (thioredoxin), all of which contributed at least 1% to the total metabolic activity.

[0185] Example 3: Pilot Study to Evaluate the Safety and Efficacy of Oral Postbiotic Therapy in Patients with Age-Related Macular Degeneration, Myopia, or Geographic Atrophy Secondary to Angioid Streaks (REVERS)

[0186] In this example, we will describe a survey that meets the following conditions: An open-label, non-randomized study in patients with macular atrophy Treatment: Postbiotics + antioxidants and vitamins in one capsule

[0187] 3.1 Research Objective The growth rate of geographic atrophy was assessed by converting the square root of the area (SQRT) measured by fundus autofluorescence (FAF) into a growth rate over the previous year. To assess the safety and tolerability of daily oral postbiotics by the incidence of ocular and non-ocular adverse events and / or clinically significant changes.

[0188] 3.2 Study design Data from patients in this study were retrospectively reviewed to characterize the progression of geographic atrophy in patients with age-related macular degeneration and in a separate group of patients with similar disease who did not receive this treatment. The study adhered to the principles of the Declaration of Helsinki and was approved by the Ethics Committee of Teknon Medical Center. Informed consent was obtained from all participants after the nature and results of the study were explained to them. An open-label, non-randomized study in patients with macular atrophy. Two groups: 10 patients with moderately rapid growth (>0.2<0.3SQRTmm / a) and known progression rates (obtained at 12-month follow-up) were included to study whether the progression rate slowed 12 months after starting treatment (14 eyes, 9 patients, 1 patient dropped out). A group of 10 patients with similar lesions received the same treatment but did not receive supplementation (13 eyes, 9 patients).

[0189] 3.2 Results 1) Oral postbiotic use was well tolerated with few side effects. 2) Progression of atrophy: After 12 months of postbiotic treatment with Sample 1, on average, lesion growth measured by SQRT showed a 26.58% reduction compared to the occurrence of the same lesions one year before treatment began. Considering only AMD patients, 6 out of 10 eyes showed a 19.87% reduction (see Figures 1 and 2).

[0190] Measurements were performed on two independent gradients using semi-automated region-finding software from Spectralis Heidelberg.

[0191] A second group of eyes with similar lesions but not treated did not show the delayed progression expected from the natural history (see Figure 3).

[0192] Figures 4 and 5 show the SQRT area before and 12 months after the start of treatment for patients who received and did not receive postbiotics, and a 26.58% deceleration and a 4.5% acceleration, respectively, were observed in accordance with the natural course.

[0193] 3.4. Conclusions of this study In this pilot study, 12 months of postbiotic treatment resulted in an average 26.58% reduction in lesion growth measured by SQRT compared to the growth of the same lesions in the year prior to treatment, compared with a 4.5% increase in control eyes (non-randomized).

[0194] 3.5. Potential medical and socio-economic impacts The advanced form of age-related macular degeneration (AMD) is the leading cause of blindness in patients over the age of 50 in developed countries, and there is no cure for the most common dry form. In dry AMD, there is death of retinal tissue affecting the central part of the retina, and this destroyed area grows relentlessly, gradually increasing in size over a very short period of time. Today, it is a real epidemic that is increasing exponentially due to the aging of the population.

[0195] Because this disease is a highly complex, multifactorial degenerative disorder, many clinical trials have failed to slow its progression. Currently, a drug based on monthly intravitreal injections is under FDA approval and has been shown to slow disease progression by 16 to 18% compared to control groups.

[0196] The 12-month REVERSE pilot study using oral postbiotic therapy in patients with macular atrophy demonstrated a mean reduction in progression of 26.58% compared to the previous year's progression in patients with geographically advanced AMD, and a 19.87% reduction in progression in patients with geographically advanced AMD. This reduction was not observed in patients who did not receive treatment; however, lesions grew by an average of 4.5%, closer to the linear growth rate known for these lesions. These results are in the same range of efficacy as currently investigational and approved drugs, but the medical and socioeconomic impact is significantly reduced because the treatment is oral compared to lifelong monthly injections. This would avoid the clinical complications associated with invasive, frequent, and chronic treatments, as well as the high socioeconomic costs for the healthcare system, patients, and their families associated with years of expensive, frequent, and ongoing treatment in the elderly.

[0197] Example 4: Study of the immunomodulatory activity of the postbiotic composition of the present invention

[0198] The following reagents and equipment were used during the study: - Fetal bovine serum (FBS). Gibco, 10270-106 - Hanks Salt H-2387 - L-Glutamine 200mM. Sigma, G7513 - Penicillin / Streptomycin (P / S). Gibco 15140-122 - Trypsin-EDTA (10X). Sigma T4174 - PBS (10x). Roche 11666789001 - MycoAlert® Mycoplasma Detection Kit. Lonza LT07-318 - RPMI-1640. Sigma, R0883 - Human IL-8 ELISA kit, BD, 555244 - Human TNF-alpha ELISA Kit, Invitrogen, EH3TNFA2 - Sodium dodecyl sulfate (SDS). CAS No. 151-21-3 Sigma, L4509 - E. coli lipopolysaccharide (LPS). Sigma, L3012 - PMA (Phorbol 12-myristate 13-acetate). Sigma, P1785 - β-mercaptoethanol. Sigma; 63689 - Ficoll® Paque Plus Cytiva. Sigma; GE17-1440-02 - Human IL-6 ELISA Kit. Invitrogen; KHC0062 - Automatic pipettes - Multichannel pipettes - Thermostatic bath with stirring function - Freezer - 80℃ - refrigerator - CO2 incubator - Laminar flow cabin - Varioskan Lux Microplate Reader (Fluorometer, Luminometer, Spectrophotometer) - Inverted optical microscope - Hemocytometer - Microplate Stirrer - Microcentrifuge

[0199] The postbiotic composition of the present invention was prepared fresh on the day of the assay directly in the culture medium according to the protocol described above. The composition is as follows:

[0200] Table 10. Composition of Sample 2 of the composition of the present invention. Shows the weight percentage of each bacterial species that makes up the dried bacterial lysate described herein.

[0201] [Table 10]

[0202] Immunomodulatory activity test THP-1 cells (human monocyte cell line, ATCC®) THP-1 cells (TIB-202) were stored in a culture bank at Gaiker and were confirmed to be free of contaminating mycoplasma immediately after thawing. THP-1 cells are grown in suspension at 37°C in a humidified atmosphere of 5% CO2. Cells were stored in culture medium (RPMI + 10% FBSi + 50 μM β-mercaptoethanol) for a minimum of 2 weeks before experimentation. Cells were replated before reaching 80% confluence (cell / volume concentration).

[0203] 1x10 24 hours before immunomodulatory activity testing 5 To differentiate monocytes into macrophages, a cell suspension of 1000 cells / mL was prepared in a 96-well plate in the presence of 0.31 μg / mL PMA. x10 4 Cells were dispensed per well and the plates were incubated at 37°C, 5% CO2 for 24 hours.

[0204] For the immunomodulatory assay, PMA-differentiated THP-1 cells were cultured with 15.63 μg / mL of the postbiotic composition of the present invention in the presence of the inflammatory stimulus lipopolysaccharide (LPS) (20 μg / mL) for 24 hours. Non-inflammatory cells (cells exposed to the culture medium without any stimulus) served as a negative control. IL-6 secretion from the cells was measured by ELISA according to the manufacturer's instructions. After the exposure period, cell supernatants were collected and analyzed. IL-6 has been reported in the latest technology to play an important role in the progression of AMD: Droho, S., Cuda, CM, Perlman, H. et al. Macrophage-derived interleukin-6 is necessary and sufficient for choroidal angiogenesis.Sci Rep 11, 18084 (2021). https: / / doi.org / 10.1038 / s41598-021-97522-x

[0205] [Table 11]

[0206] A decrease in IL-6 levels was observed compared to LPS alone, providing insight into the molecular mechanism of the therapeutic effect of the postbiotic composition of the present invention on the treatment of age-related macular degeneration.

[0207] Example 5: Pilot study to evaluate the safety and efficacy of oral postbiotic therapy using different compositions

[0208] The composition of subject composition Sample 2 of the present invention was also tested in patients in a manner similar to that of Example 3, and the results observed were very similar.

Claims

1. A composition for oral administration comprising a lysate of a probiotic microorganism in the form of a dry powder, in an amount as a weight percentage relative to the total: - 15% to 20% of bacterial lysates of the genus Bacillus, - 15% to 35% of bacterial lysate of the genus Lactobacillus, - 1.5% to 88% of Streptococcus genus bacterial lysates, 45% to 60% of bacterial lysates of Saccharomyces genus, and - 1.5% to 8% of Bifidobacterium lysate, and used in the prevention and / or treatment of degenerative eye diseases.

2. 2. A composition for use according to claim 1, comprising, in an amount as a weight percentage of the total of the melt of the composition: - about 16% of the bacterial lysate of said Bacillus genus; - about 5% of the bacterial lysate of the Bifidobacterium genus; - about 21% of the bacterial lysate of the Lactobacillus genus, - about 55% of said yeast lysate of the genus Saccharomyces; about 3% of the bacterial lysate of said Streptococcus spp. A composition comprising a lysate of a probiotic microorganism.

3. 10. A composition for use according to any of the preceding claims, wherein the bacterial lysate of the genus Bacillus is a species selected from the group consisting of Bacillus coagulans, Bacillus licheniformis, Bacillus mesentericus, Bacillus subtilis, Bacillus clausiai, Bacillus paralicheniformis, and combinations thereof.

4. 10. A composition for use according to any of the preceding claims, wherein the bacterial lysate of the genus Bifidobacterium is a species selected from the group consisting of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis subsp. lactis, Bifidobacterium infantis, Bifidobacterium animalis, and combinations thereof.

5. 10. A composition for use according to any preceding claim, wherein the bacterial lysate of the Lactobacillus genus is a species selected from the group consisting of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus kefiri, and combinations thereof.

6. 10. A composition for use according to any preceding claim, wherein the bacterial lysate of the genus Saccharomyces is a species selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces boulardii, and combinations thereof.

7. 10. A composition for use according to any preceding claim, wherein the Streptococcus bacterial lysate is a species selected from the group consisting of Streptococcus salivarius, Streptococcus thermophilus, and combinations thereof.

8. 10. A composition for use according to any of the preceding claims, comprising at least one additional ingredient selected from the group consisting of vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B9, vitamin B12, vitamin C, vitamin E, zeaxanthin, hydroxytyrosol, omega-3 fatty acid (DHA), glutathione, copper, selenium, zinc, lutein, and combinations thereof.

9. A composition for use according to any of the preceding claims, comprising, as percentages by weight: - 4% to 6% of proteins from the lysate of bacteria of the genus Bacillus; - 0.02% to 0.10% of proteins from the lysate of bacteria of the genus Bifidobacterium; - 3% to 8.50% of proteins from the bacterial lysate of the Lactobacillus genus; - 70% to 99.00% of the protein from the bacterial lysate of said Saccharomyces genus; - 0.15% to 0.90% of protein from the bacterial lysate of said Streptococcus genus; The composition includes a relative content in the range of:

10. 10. A composition for use according to any preceding claim, to be administered orally in an amount of 100 mg to 400 mg, one to six times per day.

11. A pharmaceutical composition comprising an effective pharmaceutical amount of the composition according to any of the preceding claims and a pharmaceutically acceptable excipient for use in the prevention and treatment of degenerative eye diseases.

12. The composition for use according to any one of claims 1 to 9, and the pharmaceutical composition for use according to any one of claims 10 to 11, wherein the ocular degenerative disease is age-related macular degeneration.

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