Methods to improve eyesight

Abstract

The present disclosure relates to a method of improving vision in a subject suffering from an ocular disease, comprising administering to the subject a composition comprising mesenchymal progenitor or stem cells (MLPSCs) in an amount sufficient to improve vision.

Description

[Technical Field]

[0001] This disclosure relates to cell therapy products, including mesenchymal progenitor cells or stem cells, for improving vision. [Background technology]

[0002] As a complex sensory organ of the body, the eye can experience numerous diseases and other adverse conditions that affect its ability to function normally. Many of these conditions are related to damage or degeneration of specific eye cells and the tissues composed of those cells. For example, diseases and degenerative conditions of the optic nerve and retina are major causes of blindness worldwide. Damage or degeneration of the cornea, lens, and related eye tissues represent another significant cause of vision loss. The retina contains seven alternating layers of cells and processes that convert light signals into nerve signals. In many disorders, the photoreceptors of the retina and the adjacent retinal pigment epithelium (RPE) form functional units that become unbalanced due to gene mutations or environmental conditions (including age). This leads to the loss of photoreceptors: apoptosis or secondary degeneration, which can lead to progressive deterioration of vision, and in some cases, blindness (see overview, e.g., Lund, R. D et al., 2001, Progress in Retinal and Eye Research 20:415-449). Two classes of eye disorders that fit this pattern are age-related macular degeneration (AMD) and retinitis pigmentosa (RP). [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] International Publication No. 1994 / 026877 [Patent Document 2] U.S. Patent No. 5173414 [Patent Document 3] U.S. Patent No. 5139941 [Patent Document 4] International Publication No. 92 / 01070 [Patent Document 5] International Publication No. 93 / 03769 [Patent Document 6] U.S. Patent No. 5,837,539 [Non-patent literature]

[0004] [Non-Patent Document 1] Lund, R. D et al., 2001, Progress in Retinal and Eye Research 20:415~449 [Non-Patent Document 2] "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.) [Non-Patent Document 3] Mangione et al., Arch Ophthamol. 116:1496~1505 (1998) [Non-Patent Document 4] Mangione et al., Arch Ophthamol. 119:1050~1058 (2001) [Overview of the project] [Problems that the invention aims to solve]

[0005] Therapeutic agents containing anti-VEGF agents such as Lucentis® are used to treat conditions such as wet neovascularization (AMD), where excessive VEGF production leads to abnormal leaky vascular structures and accumulation of extravascular fluid. However, there is a need for therapeutic measures that are effective in directly improving the visual acuity of patients with optic nerve diseases and degenerative conditions. [Means for solving the problem]

[0006] The present disclosure relates to the use of a commercially available ex vivo expanded allogeneic mesenchymal progenitor or stem cell (MLPSC) product for improving vision in patients suffering from an eye disorder.

[0007] Thus, the present disclosure provides a method of improving vision in a subject suffering from an eye disease, the method comprising administering to the subject a composition comprising mesenchymal progenitor or stem cells (MLPSC) in an amount sufficient to improve vision.

[0008] In one embodiment, the eye disease is related to inflammation or degeneration of the optic nerve.

[0009] In another embodiment, the eye disease is related to inflammation or degeneration of photoreceptors of the optic nerve.

[0010] In another embodiment, the subject has been previously treated with an anti-VEGF agent to reduce angiogenesis, abnormal leaky vasculature, and accumulation of extravascular fluid in the eye tissue.

[0011] In another embodiment, the subject has been treated with monthly dosing of an anti-VEGF agent for at least 1 month, or at least 2 months, or at least 3 months.

[0012] In another embodiment, the agent is an anti-VEGF antibody or a fragment thereof. For example, the anti-VEGF agent can be Lucentis®.

[0013] In another embodiment, the mesenchymal progenitor or stem cells are isolated by immunoselection.

[0014] In one embodiment, the isolated cell population comprises cultured-expanded mesenchymal progenitor or stem cells. In an alternative embodiment, the isolated cell population comprises freshly isolated mesenchymal progenitor or stem cells.

[0015] In one embodiment, MLPSCs are isolated by immunoselection. In one embodiment, cells are immunoselected for TNAP expression. In one embodiment, immunoselected cells co-express TNAP and STRO-1. In one embodiment, immunoselected cells co-express TNAP and STRO-1 bright It co-expresses. In one embodiment, immunoselected cells are cultured and expanded before administration.

[0016] In one embodiment, MLPSCs are mesenchymal stem cells. In one embodiment, the mesenchymal stem cells are cultured and expanded before administration.

[0017] In one embodiment, MLPSCs constitute at least about 5%, or at least about 10%, or at least about 20%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or at least about 95%, or 100% of the total cell population of the composition.

[0018] In one embodiment, the composition comprises MLPSC and a cryopreservative.

[0019] In one embodiment, the cryopreservative in the composition is DMSO or Profreeze®.

[0020] In one embodiment, the composition contains MLPSC in 42.5% (v / v) Profreeze® / 50% αMEM (v / v) / 7.5% (v / v) DMSO.

[0021] In another embodiment, MLPSCs are administered to subjects in doses of less than 350,000 cells, or less than 250,000 cells, or less than 100,000 cells, or less than 95,000 cells, or less than 90,000 cells, or less than 80,000 cells, or less than 75,000 cells, or less than 70,000 cells.

[0022] In another embodiment, MLPSCs are administered to subjects at doses of less than 100,000 cells per mL of vitreous fluid, or less than 75,000 cells per mL of vitreous fluid, or less than 50,000 cells per mL of vitreous fluid, or less than 25,000 cells per mL of vitreous fluid, or less than 20,000 cells per mL of vitreous fluid.

[0023] In another embodiment, MLPSCs are administered to subjects at a dose of approximately 24,500 MPCs per 1 mL of vitreous fluid.

[0024] In another embodiment, MLPSC is administered as a single dose.

[0025] In another embodiment, MLPSCs are administered intravitreously. For example, MLPSCs may be administered by intravitreous injection.

[0026] In another embodiment, administration of MLPSC results in an improvement of at least 10 points from baseline in the composite NEI VFQ-25 score over a period of at least 3 months, or at least 6 months, or at least 12 months, or at least 18 months, or at least 24 months.

[0027] In another embodiment, administration of MLPSCs results in a reduction of photocoherence tomography (OCT) over a period of 3 months. [Brief explanation of the drawing]

[0028] [Figure 1]Figure 1A is from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.). ANCHOR: Least squares mean change in NEI VFQ-25 score for patients who gained ≥15 letters, patients who gained or lost <15 letters, and patients who lost ≥15 letters, relative to the overall composite score (1A) at 12 months. Error bars represent 95%. Figure 1B is from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.). ANCHOR: Least squares mean change in NEI VFQ-25 score for three pre-specified subscales at 12 months: patients gaining ≥15 letters, patients gaining or losing <15 letters, and patients losing ≥15 letters, for nearby activity (1B). Error bars represent 95%. Figure 1C is taken from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.).ANCHOR: Least squares mean change in NEI VFQ-25 score for three pre-specified subscales over 12 months: patients gaining ≥15 letters, patients gaining or losing <15 letters, and patients losing ≥15 letters, for distant activity (1C). Error bars represent 95%. Figure 1D is from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.). ANCHOR: Least squares mean change in NEI VFQ-25 score over 12 months for three pre-specified subscales: visual acuity-specific dependency (1D) for patients gaining ≥15 letters, patients gaining or losing <15 letters, and patients losing ≥15 letters. Error bars represent 95%. [Figure 2]Figure 2A is from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.). MARINA: Least squares mean change from baseline in NEI VFQ-25 score for the overall composite score (2A) at 12 months for patients who gained ≥15 letters, patients who gained or lost <15 letters, and patients who lost ≥15 letters. Error bars represent the 95% confidence interval of the mean. Figure 2B is from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.). MARINA: Least squares mean change from baseline in NEI VFQ-25 score for three pre-specified subscales at 12 months: nearby activity (2B) for patients gaining ≥15 letters, patients gaining or losing <15 letters, and patients losing ≥15 letters. Error bars represent the 95% CI of the mean. Figure 2C is taken from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.).MARINA: Least squares mean change from baseline in NEI VFQ-25 score for three pre-specified subscales at 12 months: patients gaining ≥15 letters, patients gaining or losing <15 letters, and patients losing ≥15 letters for distant activity (2C). Error bars represent the 95% CI of the mean. Figure 2D is from "Responsiveness of NEI VFQ-25 to Changes in Visual Acuity in Neovascular AMD: Validation Studies from Two Phase 3 Clinical Trials" (Invest. Opthalmol. Vis. Sci. 2009;50(8)3629~3635. doi:10.1167 / iovs.08-3225.). MARINA: Least squares mean change from baseline in NEI VFQ-25 score for three pre-specified subscales over 12 months: for patients gaining ≥15 letters, patients gaining or losing <15 letters, and patients losing ≥15 letters, for visual acuity-specific dependency (D2). Error bars represent 95% confidence intervals of the mean. [Figure 3] This figure shows the change in neovascular thickness measured by optical coherence tomography (OCT) after three monthly Lucentis® injections and subsequent treatment with either a single intravitreous MPC injection or placebo. [Figure 4] Results regarding the effectiveness of visual acuity: This figure shows the median values ​​for patients treated with Lucentis® alone and Lucentis® + MPC. [Figure 5] This figure shows that a single intravitreal MPC injection in patients treated with Lucentis® resulted in a significant improvement in NEI VFQ-25. [Modes for carrying out the invention]

[0029] General implementation methods and definitions Throughout this specification, unless otherwise specified or contextually indicated, references to a single process, composition of a substance, group of processes, or group of compositions of a substance shall encompass one or more of those processes (i.e., one or more), a composition of a substance, group of processes, or group of compositions of a substance.

[0030] Those skilled in the art will understand that the disclosures described herein are sensitive to variations and modifications other than those specifically described. It should be understood that this disclosure includes all such variations and modifications. This disclosure also includes all, individually or collectively, the processes, features, compositions, and compounds referred to or indicated herein, and any combination or any two or more of the aforementioned processes or features.

[0031] This disclosure is intended for illustrative purposes only and is not limited in scope by the specific embodiments described herein. Functionally equivalent products, compositions, and methods are clearly within the scope of this disclosure.

[0032] Any example disclosed herein shall be construed as applying mutatis mutandis to any other example unless otherwise specified.

[0033] Unless otherwise clearly defined, all technical and scientific terms used herein shall be construed to have the same meaning as that commonly understood by those skilled in the art (e.g., cell culture, molecular genetics, stem cell differentiation, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0034] Unless otherwise noted, the stem cell, cell culture, and surgical techniques used in this disclosure are standard procedures well known to those skilled in the art. Such techniques are described and explained throughout the literature in sources such as Perbal, 1984; Sambrook & Green, 2012; Brown, 1991; Glover & Hames, 1995 and 1996; Ausubel, 1987 (including all revisions to date); Harlow & Lane, 1988; and Coligan et al., 1991 (including all revisions to date).

[0035] As used herein and in the appended claims, the singular and singular terms “a,” “an,” and “the” include, for example, multiple subjects unless the content explicitly indicates otherwise.

[0036] The term "subject" as used herein refers to mammals, including humans and non-human animals. More specifically, mammals are humans. Terms such as "subject," "patient," or "individual" are interchangeable terms in this disclosure depending on the context. In certain examples, the subject may be an adult or a child (pediatric) subject.

[0037] "Effective dose" means the minimum effective amount in the required dosage and time to achieve the desired therapeutic or prophylactic outcome. An effective dose may be provided in one or more doses. In some examples of this disclosure, the term "effective dose" is used to refer to the amount required to carry out treatment for any disease or condition as described above. The effective dose may vary depending on the disease or condition being treated, and also depending on body weight, age, racial background, sex, health status and / or physical condition, and other factors related to the mammal being treated. Typically, the effective dose falls within a relatively broad range (e.g., a range of "dosages") that can be determined by routine trials and experiments by a practicing physician. The effective dose may be administered once or several times in a single dose or in repeated doses over a period of treatment.

[0038] The term "and / or," for example "X and / or Y," shall be understood to mean either "X and Y" or "X or Y," and shall be interpreted as providing explicit support for both meanings or either meaning.

[0039] As used herein, the term “about” means, unless otherwise stated, + / - 10%, more preferably + / - 5%, of the specified value.

[0040] Throughout this specification, the terms “comprise,” or variations such as “comprises,” or “comprising,” are understood to mean encompassing the element, integer, or process, or group of elements, integers, or processes described, but not to mean excluding any other element, integer, or process, or group of elements, integers, or processes.

[0041] Mesenchymal progenitor cells or stem cells As used herein, the term “mesenchymal progenitor cell or stem cell” means an undifferentiated pluripotent cell that has the ability to regenerate while maintaining pluripotency and to differentiate into any of several cell types of mesenchymal origin, such as osteoblasts, chondrocytes, adipocytes, stromal cells, fibroblasts, and tendons, or non-mesoderm origin, such as hepatocytes, nerve cells, and epithelial cells.

[0042] The term “mesenchymal progenitor cell or stem cell” includes both parent cells and their undifferentiated offspring. This term also includes mesenchymal progenitor cells (MPCs), pluripotent stromal cells, mesenchymal stem cells, perivascular mesenchymal progenitor cells, and their undifferentiated offspring.

[0043] Mesenchymal progenitor cells or stem cells can be autologous, allogeneic, allogeneic, syngeneic, or isogeneic. Autologous cells are isolated from the same individual from which they are re-transplanted. Allogeneic cells are isolated from an allogeneic donor. Allogeneic cells are isolated from a donor of a different species. Syngeneic or isogeneic cells are isolated from genetically identical organisms, such as twins, clones, or highly inbred research animal models.

[0044] Mesenchymal progenitor cells or stem cells are primarily found in the bone marrow, but have also been shown to be present in a variety of host tissues, including, for example, umbilical cord blood and umbilical cord, adult peripheral blood, adipose tissue, trabeculae, and dental pulp.

[0045] Mesenchymal progenitor cells or stem cells can be isolated from host tissue and enriched by immunoselection. For example, bone marrow aspirates from a subject can be further treated with antibodies against STRO-1 or TNAP to enable selection of mesenchymal progenitor cells or stem cells. In one example, mesenchymal progenitor cells or stem cells can be enriched using the STRO-1 antibody described in Simmons & Torok-Storb, 1991.

[0046] STRO-1+ cells are found in bone marrow, blood, dental pulp cells, adipose tissue, skin, spleen, pancreas, brain, kidney, liver, heart, retina, hair follicles, intestines, lungs, lymph nodes, thymus, bone, ligaments, tendons, skeletal muscle, dermis, and periosteum, and can differentiate into germline cells such as mesoderm and / or endoderm and / or ectoderm. Therefore, STRO-1+ cells can differentiate into numerous cell types, including, but not limited to, adipose tissue, bone, cartilage, elastic tissue, muscle, and fibrous connective tissue. The specific lineages involved and differentiation pathways through which these cells invade depend on various influences from mechanical factors such as growth factors, cytokines, and / or local microenvironmental conditions established by the host tissue, and / or endogenous bioactive factors.

[0047] The term “enriched,” as used herein, describes a population of cells in which the proportion of one particular cell type or a number of particular cell types is increased compared to an untreated population of cells (e.g., cells in their natural environment). For example, a population enriched for STRO-1+ cells contains at least about 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 50%, or 75% STRO-1+ cells. In this regard, the term “population of cells enriched for STRO-1+ cells” is used to explicitly support the term “population of cells containing X% STRO-1+ cells,” where X% is the percentage described herein. STRO-1+ cells may, in some examples, form clonal colonies, e.g., CFU-F (fibroblasts) or subsets thereof (e.g., 50%, 60%, 70%, 70%, 90%, or 95%) and may possess this activity. For example, a population enriched with TNAP+ cells contains at least about 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 50%, or 75% TNAP+ cells. In this regard, the term “population of cells enriched with TNAP+ cells” is used to explicitly support the term “population of cells containing X% TNAP+ cells,” where X% is the percentage described herein. For example, a population enriched with STRO-1+ and TNAP+ cells contains at least about 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 50%, or 75% STRO-1+ and TNAP+ cells. In this regard, the term “population of cells enriched with STRO-1+ and TNAP+ cells” is used to explicitly support the term “population of cells containing X% STRO-1+ and TNAP+ cells,” where X% is the percentage specified herein.

[0048] In one example, a cell population is enriched from a cell preparation containing STRO-1+ cells in a selectable form. In this regard, the term “selectable form” is understood to mean that the cells express a marker (e.g., a cell surface marker) that enables the selection of STRO-1+ cells. The marker may, but does not necessarily, be STRO-1. For example, cells expressing STRO-2 and / or STRO-3 (TNAP) and / or STRO-4 and / or VCAM-1 and / or CD146 and / or 3G5 (e.g., MPC) as described and / or illustrated herein also express STRO-1 (and STRO-1 bright (This is possible). Therefore, the indication that cells are STRO-1+ does not mean that cells are selected by STRO-1 expression. In one example, cells are selected based on at least STRO-3 expression, for example, they are STRO-3+(TNAP+).

[0049] References to the selection of cells or populations thereof do not necessarily require selection from a specific tissue source. As described herein, STRO-1+ cells may be selected from, isolated from, or enriched from a wide variety of sources. Nevertheless, in some examples, these terms support the selection of STRO-1+ cells, or vascularized tissue, or tissue containing pericytes (e.g., STRO-1+ pericytes), or any one or more tissues including the tissues described herein.

[0050] In one example, the mesenchymal progenitor cells or stem cells of the present disclosure express one or more markers individually or collectively selected from the group consisting of TNAP+, VCAM-1+, THY-1+, STRO-2+, STRO-4+(HSP-90β), CD45+, CD146+, and 3G5+.

[0051] "Individually" means that this disclosure separately encompasses the markers or groups of markers described herein, and that even though individual markers or groups of markers may not be separately listed herein, the appended claims may define such markers or groups of markers separately and separately from one another.

[0052] "Collectively" means that this disclosure encompasses any number or combination of the markers or groups of markers described herein, and that even if such a number or combination of markers or groups of markers is not specifically enumerated herein, the appended claims may define such combinations or subcombinations separately and apart from any other combination of markers or groups of markers.

[0053] Cells referred to as "positive" for a given marker may express the marker at low (lo, dim, or dull), intermediate (median), or high (bright, bri) levels, depending on the extent to which the marker is present on the cell surface. These terms relate to the intensity of fluorescence or other markers used in cell sorting processes or flow cytometry analysis of cells. The distinction between low (lo, dim, or dull), intermediate (median), and high (bright, bri) is understood in relation to the marker used on the particular cell population being sorted or analyzed. Cells referred to as "negative" for a given marker do not necessarily mean that the cell is completely absent. This term means that the marker is expressed by the cell at a relatively very low level, producing a very low signal when detectably labeled, or being undetectable above background levels, for example, levels detected using an isotype control antibody.

[0054] As used herein, the term "bright" or "bri" refers to a marker on the cell surface that generates a relatively high signal when detectably labeled. Without wishing to be bound by theory, it is proposed that "bright" cells express more target marker proteins (e.g., the antigen recognized by the STRO-1 antibody) than other cells in the sample. For example, STRO-1 bri cells, when labeled with a STRO-1 antibody conjugated to FITC as determined by fluorescence-activated cell sorting (FACS) analysis, produce a larger fluorescent signal than non-bright cells (STRO-1 lo / dim / dull / intermediate / median ). In one example, mesenchymal progenitor cells or stem cells are isolated from bone marrow and enriched by selection of STRO-1+ cells. In this example, "bright" cells constitute at least about 0.1% of the most brightly labeled bone marrow mononuclear cells included in the starting sample. In other examples, "bright" cells constitute at least about 0.1%, at least about 0.5%, at least about 1%, at least about 1.5%, or at least about 2% of the most brightly labeled bone marrow mononuclear cells included in the starting sample. In one example, STRO-1 bright cells have a high expression of STRO-1 surface expression that is 2 logs greater compared to "background", i.e., cells that are STRO-1-. In contrast, STRO-1 lo / dim / dull and / or STRO-1 intermediate / median cells show expression of higher STRO-1 surface expression on a scale of less than 2 logs, typically about 1 log or less than "background".

[0055] In one example, STRO-1+ cells are STRO-1 bright [[ID=十六]]In one example, STRO-1 bright cells are preferentially enriched compared to STRO-1 lo / dim / dull or STRO-1 intermediate / median cells.

[0056] [[ID=二十五]] In one example, STRO-1 brightThe cells are further characterized by being one or more of the following: TNAP+, VCAM-1+, THY-1+, STRO-2+, STRO-4+ (HSP-90β), and / or CD146+. For example, cells are selected for one or more of the aforementioned markers and / or shown to express one or more of the aforementioned markers. In this regard, cells shown to express a marker do not need to be specifically tested; rather, previously enriched or isolated cells can be tested and then reasonably presumed to also express the same marker.

[0057] For example, STRO-1 bright The cells are perivascular mesenchymal progenitor cells, as defined in International Publication No. 2004 / 85630, characterized by the presence of the perivascular marker 3G5.

[0058] As used herein, the term "TNAP" is intended to encompass all isoforms of tissue-nonspecific alkaline phosphatase. For example, the term includes the liver isoform (LAP), the bone isoform (BAP), and the kidney isoform (KAP). In one example, TNAP is BAP. In one example, TNAP refers to a molecule capable of binding to a STRO-3 antibody produced by a hybridoma cell line deposited with ATCC on December 19, 2005, under the provisions of the Budapest Convention under depositary number PTA-7282.

[0059] Furthermore, in one example, STRO-1+ cells can induce clonal CFU-F.

[0060] In one example, a significant proportion of STRO-1+ cells can differentiate into at least two different germlines. Non-limiting examples of lineages in which cells may be involved include bone progenitor cells; hepatocyte precursors that are multipotent for cholangiocarcinoma and hepatocytes; neuron limiting cells that can produce glial cell precursors that progress to oligodendrocytes and astrocytes; neuronal precursors that progress to neurons; cardiomyocyte and cardiomyocyte precursors; and glucose-responsive insulin-secreting pancreatic beta cell lineages. Other lineages, but not limited to, include odontoblasts, dentin-producing cells and chondrocytes, as well as: dermal cells such as retinal pigment epithelial cells, fibroblasts, keratinocytes, dendritic cells, hair follicle cells, renal duct epithelial cells, smooth muscle and skeletal muscle cells, testicular progenitor cells, vascular endothelial cells, tendons, ligaments, cartilage, adipocytes, fibroblasts, bone marrow stroma, cardiomyocytes, smooth muscle, skeletal muscle, pericytes, blood vessels, epithelium, glia, nerve cells, astrocytes and oligodendrocyte precursors.

[0061] In one example, mesenchymal progenitor cells or stem cells are mesenchymal stem cells (MSCs). MSCs may be a homogeneous composition or a mixed cell population enriched with MSCs. A homogeneous MSC composition can be obtained by culturing adherent bone marrow or periosteal cells, and MSCs may be identified by specific cell surface markers identified by specific monoclonal antibodies. A method for obtaining an MSC-enriched cell population using plastic adhesion techniques is described, for example, in U.S. Patent No. 5,486,359. MSCs prepared by conventional plastic adhesion isolation rely on the nonspecific plastic adhesion properties of CFU-F. Alternative sources of MSCs include, but are not limited to, blood, skin, umbilical cord blood, muscle, fat, bone, and perichondrium.

[0062] Mesenchymal progenitor cells or stem cells can be cryopreserved before administration to the target organism.

[0063] In a preferred embodiment of the present invention, mesenchymal progenitor cells or stem cells are obtained from a master cell bank derived from mesenchymal progenitor cells or stem cells enriched from the bone marrow of healthy volunteers. The use of mesenchymal progenitor cells or stem cells derived from such a source is particularly advantageous for subjects who can act as mesenchymal progenitor cell or stem cell donors, or who require immediate treatment and have no suitable family members available who are at high risk of relapse, disease-related decline, or death during the time required to generate mesenchymal progenitor cells or stem cells.

[0064] The inventors demonstrate that the mesenchymal progenitor cells of this disclosure possess unexpectedly high efficacy in inhibiting T cell proliferation after cryopreservation and thawing. In contrast, previous disclosures have taught that cryopreserved mesenchymal stem cells lose their immunosuppressive properties after thawing (Francois et al., 2012; Chinnadurai et al., 2016).

[0065] Isolated or enriched mesenchymal progenitor cells or stem cells can be expanded by culture ex vivo or in vitro. As will be understood by those skilled in the art, isolated or enriched mesenchymal progenitor cells or stem cells can be cryopreserved, thawed, and then further expanded by culture ex vivo or in vitro.

[0066] Cultured mesenchymal progenitor cells or stem cells are phenotypically different from in vivo cells. For example, in one embodiment, they express one or more of the following markers: CD44, NG2, DC146, and CD140b.

[0067] Cultured mesenchymal progenitor cells or stem cells are biologically different from cells in vivo and, in vivo, exhibit a higher proliferation rate compared to mostly non-periodic (resting) cells.

[0068] In one example, a cell population enriched with mesenchymal progenitor cells or stem cells was cultured in serum-supplemented culture medium, such as Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 2 mM glutamine, at a concentration of approximately 6,000–7,000 viable cells / cm³. 2 The cells are seeded and allowed to adhere to the culture vessel overnight at 37°C and 20% O2. In one embodiment, the cells are approximately 6000, 6100, 6200, 6300, 6400, 6500, 6600, 6700, 6800, 6810, 6820, 6830, 6840, 6850, 6860, 6870, 6880, 6890, 6890, 6900, 6910, 6920, 6930, 6940, 6970, 6980, 6990, or 7000 viable cells / cm². 2 Preferably about 6850-6860 viable cells / cm² 2 The cells are then seeded. Next, the culture medium is replaced, and the cells are cultured at 37°C and 5% O2 for a total of 68-72 hours before co-culturing with T cells and determining the amount of IL-2Rα expressed by T cells.

[0069] Composition and administration Compositions comprising mesenchymal progenitor cells or stem cells may be prepared in a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier,” as used herein, refers to a composition of substances that facilitates the storage, administration, and / or maintain the biological activity of mesenchymal progenitor cells or stem cells.

[0070] In one example, the carrier does not cause significant local or systemic adverse effects in the recipient. A pharmaceutically acceptable carrier may be solid or liquid. Useful examples of pharmaceutically acceptable carriers include, but are not limited to, diluents, solvents, surfactants, excipients, suspensions, buffers, lubricants, adjuvants, vehicles, emulsifiers, absorbents, dispersions, coatings, stabilizers, protective colloids, adhesives, thickeners, thixotropic agents, penetrating agents, chelating agents, scaffolds, isotonic and absorption retardants, which do not affect the viability and activity of mesenchymal progenitor cells or stem cells. The selection of a suitable carrier is within the scope of the art of the art.

[0071] The compositions of this disclosure can be conveniently provided in unit dosage forms and can be prepared by any method well known in the art. The term “unit dosage form” as used herein means a physically discrete unit suitable as a single dose for the subject to be treated; each unit contains a predetermined amount of the active compound calculated to produce a desired therapeutic or prophylactic effect in relation to the pharmaceutical carrier. The dose of mesenchymal progenitor cells or stem cells may vary depending on factors such as the disease state, age, sex, and weight of the subject to be treated.

[0072] The term "subject" refers to animals, preferably mammals, including non-primates (e.g., cattle, pigs, horses, cats, dogs, rats, or mice) and primates (e.g., monkeys or humans). In preferred embodiments, the subject is human.

[0073] Mesenchymal progenitor cells or stem cells constitute at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the cell population of the composition.

[0074] The compositions of this disclosure can be cryopreserved. Cryopreservation of mesenchymal progenitor cells or stem cells can be carried out using slow cooling methods or “fast” freezing protocols known in the art. Preferably, the cryopreservation method maintains similar phenotypes, cell surface markers, and proliferation rates of the cryopreserved cells compared to unfrozen cells.

[0075] The cryopreservation composition may include a cryopreservation solution. The pH of the cryopreservation solution is typically 6.5 to 8, preferably 7.4.

[0076] The cryopreservation solution may contain a sterile, non-pyrogenic isotonic solution, such as PlasmaLyteA®. 100 mL of PlasmaLyteA® contains 526 mg of sodium chloride, USP(NaCl); 502 mg of sodium gluconate (C6H11NaO7); 368 mg of sodium acetate trihydrate, USP(C2H3NaO2·3H2O); 37 mg of potassium chloride, USP(KCl); and 30 mg of magnesium chloride, USP(MgCl2·6H2O). It does not contain antimicrobial agents. The pH is adjusted with sodium hydroxide. The pH is 7.4 (6.5-8.0).

[0077] The cryopreservation solution may contain Profreeze®. The cryopreservation solution may additionally or alternatively contain culture medium.

[0078] To facilitate freezing, cryoprotective agents, such as dimethyl sulfoxide (DMSO), are typically added to the cryopreservation solution. Ideally, the cryoprotective agent should be non-toxic to cells and patients, non-antigenic, chemically inactive, provide high viability after thawing, and allow transplantation without washing. However, DMSO, the most commonly used cryoprotective agent, exhibits some cytotoxicity. Hydroxyretil starch (HES) can be used as a substitute or in combination with DMSO to reduce the cytotoxicity of the cryopreservation solution.

[0079] The cryopreservation solution may contain one or more of DMSO, hydroxyethyl starch, human serum components, and other protein bulking agents. For example, the cryopreservation solution may contain about 5% human serum albumin (HSA) and about 10% DMSO. The cryopreservation solution may further contain one or more of methylcellulose, polyvinylpyrrolidone (PVP), and trehalose.

[0080] In one embodiment, the cells are suspended in 42.5% Profreeze® / 50% αMEM / 7.5% DMSO and cooled in a freezer at a controlled rate.

[0081] The cryopreserved composition can be thawed and administered directly to the target. Alternatively, the cryopreserved composition can be thawed, and the mesenchymal progenitor cells or stem cells can be resuspended in an alternative carrier before administration.

[0082] Genetically modified cells In one embodiment, the mesenchymal progenitor cells or stem cells are not genetically modified. In one embodiment, the mesenchymal progenitor cells or stem cells are genetically modified to express and / or secrete, for example, a protein of interest, such as a protein that provides therapeutic and / or prophylactic benefits.

[0083] Methods for genetically modifying cells are apparent to those skilled in the art. For example, nucleic acids to be expressed in cells are operably ligated to promoters for inducing their expression in cells. For example, nucleic acids are ligated to promoters operable in various target cells, such as viral promoters, e.g., CMV promoters (e.g., CMV-IE promoters) or SV-40 promoters. Additional suitable promoters are known in the art.

[0084] Preferably, nucleic acids are provided in the form of expression constructs. The term “expression construct,” as used herein, means a nucleic acid capable of operatively mediated expression in a cell (e.g., a reporter gene and / or a counter-selectable reporter gene). In connection with this disclosure, it should be understood that expression constructs may or may include plasmids, bacteriophages, phagemids, cosmids, viral subgenomes or genomic fragments, or other nucleic acids capable of holding and / or replicating heterologous DNA in an expressible form.

[0085] Methods for constructing suitable expression constructs for carrying out the present invention are apparent to those skilled in the art and are described, for example, in Ausubel FM, 1987 (including all revisions to date); or Sambrook and Green, 2012. For example, each component of an expression construct is amplified from a suitable template nucleic acid, for example, using PCR, and then cloned into a suitable expression construct, for example, a plasmid or phagemid.

[0086] Suitable vectors for such expression constructs are known in the art and / or described herein. For example, suitable expression vectors for the methods of the present invention in mammalian cells include, for example, a pcDNA vector set (Invitrogen), a pCI vector set (Promega), a pCMV vector set (Clontech), a pM vector (Clontech), a pSI vector (Promega), a VP 16 vector (Clontech), or a pcDNA vector set (Invitrogen).

[0087] Those skilled in the art are aware of additional vectors and sources of such vectors, such as Invitrogen Corporation, Clontech, or Promega.

[0088] Means for introducing isolated nucleic acid molecules or gene constructs containing them into cells for expression are known to those skilled in the art. The techniques used for a given organism depend on known successful techniques. Means for introducing recombinant DNA into cells include, in particular, microinjection, DEAE-dextran-mediated transfection, liposome-mediated transfection such as using lipofectamine (Gibco, MD, USA) and / or cellfectin (Gibco, MD, USA), PEG-mediated DNA uptake, electroporation, and microparticle irradiation such as using DNA-coated tungsten or gold particles (Agracetus Inc., WI, USA).

[0089] Alternatively, the expression construct of the present invention is a viral vector. Suitable viral vectors are known and commercially available in the art. Conventional virus-based systems for delivering nucleic acids and incorporating them into the host cell genome include, for example, retroviral vectors, lentiviral vectors, or adeno-associated virus vectors. Alternatively, adenovirus vectors are useful for introducing nucleic acids with episomes into host cells. Viral vectors are a highly efficient and versatile method for gene transfer into target cells and tissues. Furthermore, high transduction efficiencies have been observed in many different cell types and target tissues.

[0090] For example, retroviral vectors typically contain a cis-acting long-terminal repeat (LTR) with a packaging capacity of up to 6–10 kb for exogenous sequences. A minimal cis-acting LTR is sufficient for vector replication and packaging, which is then used to incorporate the expression construct into target cells to provide long-term expression. Widely used retroviral vectors include those based on mouse leukemia virus (MuLV), gibbon leukemia virus (GaLV), simian immunodeficiency virus (SrV), human immunodeficiency virus (HIV), and combinations thereof (see, for example, International Publication No. 1994 / 026877; Buchschacher and Panganiban, 1992; Johann et al., 1992; Sommerfelt and Weiss, 1990; Wilson et al., 1989; Miller et al., 1991; Lynch et al., 1991; Miller and Rosman, 1989; Miller, 1990; Scarpa et al., 1991; Burns et al., 1993).

[0091] Various adeno-associated virus (AAV) vector systems have also been developed for nucleic acid delivery. AAV vectors can be readily constructed using techniques known in the art (see, for example, U.S. Patents 5,173414 and 5139941; International Publications 92 / 01070 and 93 / 03769; Lebkowski et al., 1988; Vincent et al., 1990; Carter, 1992; Muzyczka, 1992; Kotin, 1994; Shelling and Smith, 1994; Zhou et al., 1994).

[0092] Additional viral vectors useful for delivering the expression constructs of the present invention include, for example, those derived from poxviruses such as vaccinia viruses and avian poxviruses, or alphaviruses, or conjugate viral vectors (e.g., those described by Fisher-Hoch et al., 1989).

[0093] Those skilled in the art will understand that numerous modifications and / or alterations can be made to the embodiments described above without departing from the broad general scope of this disclosure. Therefore, these embodiments should be considered in all respects to be illustrative and not restrictive. [Examples]

[0094] (Example 1) Materials and methods Mesenchymal progenitor cells or stem cells (MLPSCs) prepared using plastic bonding technology MLPSCs were generated de novo from bone marrow as described in U.S. Patent No. 5,837,539. Approximately 80-100 ml of bone marrow was aspirated into a sterile heparin-containing syringe and sent to MDACC Cell Therapy Laboratory for MSC generation. Bone marrow mononuclear cells were isolated using ficoll-hypaque, and MLPSC expansion medium containing alpha-modified MEM (αMEM) with gentamicin, glutamine (2 mM), and 20% (v / v) fetal bovine serum (FBS) (Hyclone) was placed in two T175 flasks, each containing 50 ml. The cells were cultured at 37°C and 5% CO2 for 2-3 days, during which time non-adherent cells were removed. The remaining adherent cells were then continuously cultured until the cells reached a confluence of 70% or more (7-10 days). Subsequently, the cells were trypsinized and transferred to six T175 flasks containing expanded medium (50 ml of medium per flask).

[0095] Immunoselection of mesenchymal progenitor cells or stem cells (MLPSCs) Bone marrow (BM) was collected from healthy adult volunteers (ages 20-35). Simply put, 40 ml of BM was aspirated from the posterior iliac crest into a lithium-heparin anticoagulant-containing tube.

[0096] Bone marrow mononuclear cells (BMMNCs) were prepared by density gradient separation using Lymphoprep® (Nycomed Pharma, Oslo, Norway), which was previously described by Zannettino et al., 1998. After centrifugation at 400 × g for 30 minutes at 4°C, the buffy layer was removed with a transfer pipette and washed three times in "HHF," which consists of Hank's equilibrium salt solution (HBSS; Life Technologies, Gaithersburg, MD) containing 5% fetal bovine serum (FCS, CSL Limited, Victoria, Australia).

[0097] STRO-3+ (or TNAP+) cells were isolated by magnetically activated cell sorting, as previously described by Gronthos & Simmons, 1995; and Gronthos, 2003. Briefly, they are approximately 1–3 × 10⁶ cells in size. 8 Each BMMMNC cell is incubated on ice for 20 minutes in a blocking buffer consisting of 10% (v / v) normal rabbit serum in HHF. The cells are incubated on ice for 1 hour with 200 μl of STRO-3 mAb in a 10 μg / ml solution in blocking buffer. Subsequently, the cells are washed twice in HHF by centrifugation at 400 × g. A 1 / 50 dilution of goat anti-mouse γ-biotin (Southern Biotechnology Associates, Birmingham, UK) is added to the HHF buffer, and the cells are incubated on ice for 1 hour. The cells are then incubated in MACS buffer (containing 1% BSA, 5 mM EDTA, and 0.01% sodium azide) as described above. 2+ and Mg 2+ Washed twice in PBS (which does not contain [specific component]), and then resuspended in 0.9 ml of MACS buffer to a final volume.

[0098] 100 μl of streptavidin microbeads (Miltenyi Biotec; Bergisch Gladbach, Germany) were added to the cell suspension and incubated on ice for 15 minutes. The cell suspension was washed twice, resuspended in 0.5 ml of MACS buffer, and then loaded onto a mini-MACS column (MS column, Miltenyi Biotec). The column was washed three times with 0.5 ml of MACS buffer to collect cells that did not bind to STRO-3 mAb (deposited in the American Type Culture Collection (ATCC) on December 19, 2005, with accession number PTA-7282 - see International Publication No. 2006 / 108229). After adding another 1 ml of MACS buffer, the column was removed from the magnet, and TNAP+ cells were isolated under positive pressure. Aliquots of cells from each fraction were stained with streptavidin-FITC, and their purity could be evaluated by flow cytometry.

[0099] (Example 2) clinical research Designing a Clinical Study The purpose of this study is to evaluate the safety and feasibility of a single intravitreal injection of 93,750 allogeneic microplastics (MPCs) in patients receiving Lucentis® treatment.

[0100] Another objective is to investigate the functional effectiveness of intravitreous microcontrollers (MPCs) in visual acuity, the combined average NIA VFQ-25, and optical coherence tomography (OCT).

[0101] Another objective is to reduce the number of anti-VEGF injections required to prevent vascular leakage.

[0102] Study Design: A Phase Ib / IIa randomized, placebo-controlled study investigating the safety and feasibility of a single intravitreal injection of allogeneic MPC in subjects with newly diagnosed neovascular AMD after three monthly injections of Lucentis® treatment.

[0103] Protocol: All patients received three monthly intravitreal Lucentis® injections. At month 4, all patients (N=9) were randomized in a 2:1 ratio to receive either a single MPC injection or placebo.

[0104] Investigational products and administration The investigational product was STRO-3 selected allogeneic MPC, derived from cultured and expanded adult bone marrow mononuclear cells that were subsequently cryopreserved. The allogeneic MPC was formulated at concentrations of 30 million and 90 million nucleated cells per 5 mL volume and cryopreserved in 7.5% dimethyl sulfoxide / 50% alpha-modified Eagle medium and 42.5% ProFreeze®.

[0105] The investigational products were stored in the gas phase of liquid nitrogen at -140°C to 196°C until ready for use. The investigational products had to be properly identified and isolated from other products.

[0106] The doses for this study were 93,750 MPCs (0.03 mL from 3 million MPCs per 1 mL bag) and 312,500 MPCs (0.05 mL from 6 million MPCs per 1 mL bag). When the dose is expressed relative to the vitreous volume of the eye, the starting dose in this study is approximately 24,500 MPCs per 1 mL of vitreous fluid.

[0107] National Academy of Ophthalmology Visual Function Questionnaire (NEI VFQ-25) The NEI VFQ-25 was developed to measure the perception of patients' vision-related functions. See, for example, Mangione et al., Arch Ophthamol. 116:1496~1505 (1998); and Mangione et al., Arch Ophthamol. 119:1050~1058 (2001).

[0108] Results published by Suner et al. in 2009 (Suner et al., Inv Ophthamol. & Visula science, 50(8):3629~3635) support the use of the NEI VFQ-25 as a response and sensory measure of vision-related functions in neovascular AMD populations. Based on data from ANCHOR (Figure 1) and MARINA (Figure 2), a 4-6 point change in the NEI VFQ-25 score represents a clinically significant change corresponding to a change in the 15 letters of BCVA.

[0109] The NEI VFQ-25 demonstrated response and sensitivity to clinically significant changes in visual acuity in the MARINA and ANCHOR clinical trials. Significant differences were observed among the three visual acuity subgroups (≥15 letter gain, <15 letter loss or gain, or ≥15 letter loss) in the composite score and three pre-specified endpoints: near activity, far activity, and visual-specific dependence. This study provides evidence that the NEI VFQ-25 responds to changes in visual acuity in patients receiving pharmacological treatment for neovascular AMD.

[0110] Results of clinical research Figure 3 shows changes in optical coherence tomography (OCT) in patients who received three monthly Lucentis® injections, followed by a single intravitreal MPC injection or placebo at 4 months. Both patient groups showed similar significant reductions in retinal thickness up to 3 months, consistent with the similar therapeutic effects of three monthly Lucentis® injections in each group, as measured by OCT. This indicates that abnormal angiogenesis and perivascular extravascular fluid accumulation associated with inflammation were significantly reduced before MPC injection. From 4 months onward, the degree of OCT changes did not differ between groups, indicating that MPC injection did not further increase abnormal angiogenesis and vascular leakage beyond the initial treatment with Lucentis®.

[0111] Figure 4 shows the efficacy results in terms of visual acuity scores from baseline. The results show the mean values ​​for patients treated with Lucentis® alone and patients treated with a single injection of Lucentis® and MPC. Compared to the Lucentis® alone group, which showed a significant deterioration in visual acuity over 12 months, the group that received a single MPC injection demonstrated maintenance of visual acuity over this period.

[0112] Figure 5 shows the NEI VFQ-25 results after a single intravitreal MPC injection in patients treated with Lucentis® compared to a control patient treated with Lucentis® who received a placebo.

[0113] These results indicate that MPC injection after Lucentis® treatment is effective in the following ways. i. Prevention and stabilization of vision loss over a 12-month period; and ii. Compared to a 3-point loss in the Lucentis® group alone, an 11-point improvement from baseline in the composite NEI VFQ-25 score in the MPC+Lucentis® group suggests that the MPC group may experience an improvement of at least 15 letters in visual acuity over 12 months, while the control group may experience the same amount of visual acuity loss.

[0114] These results demonstrate that a single MPC injection is effective in improving visual acuity compared to patients treated with Lucentis® who received placebo, in patients with neovascular AMD whose angiogenesis was adequately treated with Lucentis® and whose inflammatory thickening of the neovascular membrane caused by vascular leakage and perivascular fluid was adequately reversed. Since both groups subsequently continued to receive the same number of additional Lucentis® injections as needed, this indicates that MPC improves visual acuity by acting directly on the optic nerve, because any neovascularization or inflammation present in the eye was adequately treated with previous monthly Lucentis® injections.

[0115] [References] TIFF0007871055000001.tif205170TIFF0007871055000002.tif116170

Claims

1. A composition comprising a population of mesenchymal progenitor cells for use in a method for improving visual acuity in human subjects suffering from an eye disease, wherein the amount of mesenchymal progenitor cells in the composition is sufficient to improve visual acuity in the human subjects, and the eye disease is related to inflammation of the optic nerve.

2. The composition according to claim 1, wherein the eye disease is related to the degeneration of photoreceptors in the optic nerve.

3. The composition according to claim 1 or 2, wherein the human subject has been previously treated with an anti-VEGF antibody or a fragment thereof to reduce angiogenesis in ocular tissue.

4. The composition according to any one of claims 1 to 3, wherein the human subject is treated with monthly administration of an anti-VEGF antibody or a fragment thereof for at least one to three months.

5. The composition according to any one of claims 1 to 3, wherein the human subject is treated with monthly administration of an anti-VEGF antibody or a fragment thereof for at least two to three months.

6. The composition according to any one of claims 1 to 5, wherein the population of mesenchymal progenitor cells (MPCs) is a population of cultured MPCs.

7. The composition according to any one of claims 1 to 6, comprising a dose of less than 350,000 cells, or less than 250,000 cells, or less than 100,000 cells, or less than 95,000 cells, or less than 90,000 cells, or less than 80,000 cells, or less than 75,000 cells, or less than 70,000 cells.

8. The composition according to any one of claims 1 to 7, comprising a dose of less than 100,000 cells per 1 mL of vitreous fluid, or less than 75,000 cells per 1 mL of vitreous fluid, or less than 50,000 cells per 1 mL of vitreous fluid, or less than 25,000 cells per 1 mL of vitreous fluid, or less than 20,000 cells per 1 mL of vitreous fluid.

9. The composition according to any one of claims 1 to 7, comprising a dose of approximately 24,500 MPCs per 1 mL of vitreous fluid.

10. A composition according to any one of claims 1 to 9, formulated for administration as a single dose.

11. A composition according to any one of claims 1 to 10, formulated for intravitreous administration.

12. The composition according to any one of claims 1 to 11, wherein administration of the composition results in an improvement of at least 10 points from baseline in the composite NEI VFQ-25 score over a period of at least 3 months, or at least 6 months, or at least 12 months, or at least 18 months, or at least 24 months.

13. The composition according to any one of claims 1 to 12, wherein administration of the composition results in a reduction of photocoherence tomography (OCT) over a period of three months.

14. Use of a composition comprising a population of mesenchymal progenitor cells in the manufacture of a pharmaceutical product for improving visual acuity in human subjects suffering from an eye disease, wherein the amount of mesenchymal progenitor cells in the composition is sufficient to improve visual acuity in the human subjects, and the eye disease is related to inflammation of the optic nerve.

15. The use according to claim 14, wherein the human subject is treated with monthly administration of an anti-VEGF antibody or a fragment thereof for at least one to three months.

16. The use according to claim 14 or 15, wherein the pharmaceutical product is formulated for intravitreous administration.

17. The use according to any one of claims 14 to 16, wherein administration of the composition results in an improvement of at least 10 points from baseline in the composite NEI VFQ-25 score over a period of at least 3 months, or at least 6 months, or at least 12 months, or at least 18 months, or at least 24 months.

18. The use according to any one of claims 14 to 17, wherein administration of the composition results in a reduction of photocoherence tomography (OCT) over a period of three months.

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