Treatments for eye diseases

Abstract

The present invention provides a method for treating an eye disease such as age-related macular degeneration (AMD), diabetic retinopathy (DR), or macular edema (ME), comprising administering a composition comprising a therapeutically effective amount of beauvericin to an individual in need thereof.

Description

Technical Field

[0001] The present invention relates to a method for treating eye diseases such as age-related macular degeneration, diabetic retinopathy, or macular edema. Background Art

[0002] Age-related macular degeneration (AMD) and diabetic retinopathy (DR) are the leading causes of acquired blindness in developed countries and are characterized by pathologic posterior segment neovascularization (PSNV). Pathologic ocular angiogenesis, including PSNV, occurs as a series of events that progress from an initial stimulus to the formation of abnormal new microvessels. The primary cause of AMD and DR remains unknown, however, the production of various growth factors that promote angiogenesis appears to be a common stimulus. Pathologic ocular angiogenesis has been found in tissues and fluids of patients. After initiation of the angiogenic cascade, microvascular basement membrane and extracellular matrix are degraded, and microvascular endothelial cells proliferate and migrate. Endothelial outgrowth anastomoses to form tubes that subsequently form transluminal tracts. New microvessels often have increased vascular permeability or leakage due to a poorly developed barrier function, which can lead to tissue edema. The presence of a continuous basement membrane and normal endothelial connections between other endothelial cells and the outer envelope cells indicate differentiation toward mature microvessels; however, this differentiation process is often impaired under pathological conditions.

[0003] Macular edema (ME) is the leading cause of visual loss in diabetic patients, while preretinal neovascularization (PDR) is the leading cause of actual blindness. Diabetes is characterized by persistent hyperglycemia, which causes reversible and irreversible pathological changes in the microvasculature of various organs. Diabetic retinopathy (DR) is therefore a retinal microvascular disease that presents as a series of stages with increasing severity and worsening visual prognosis. The major risk factors for diabetic retinopathy include the duration of diabetes, glycemic control, and the presence of systemic hypertension. Diabetic retinopathy (DR) is usually classified into two major clinical stages: non-proliferative diabetic retinopathy (NPDR) and proliferative diabetic retinopathy (PDR), where the term "proliferative" refers to preretinal neovascularization as described above.

[0004] Macular degeneration, also known as age-related macular degeneration (AMD or ARMD), is a medical condition that can cause blurred or no vision in the center of the field of vision. There are usually no symptoms in the early stages; however, over time, some people's vision gradually worsens and may affect one or both eyes. There are some ways to prevent it, including exercise, a good diet, and not smoking, but there is no cure or treatment to restore vision that has been lost. Anti-VEGF (vascular endothelial growth factor) drugs injected into the eye in a wet form or, less commonly, laser coagulation or photodynamic therapy can slow progression. Supplements may slow the progression of the disease in people who already have it.

[0005] There are currently no approved pharmacological therapies for treating age-related macular degeneration (AMD), diabetic retinopathy (DR), and / or macular edema. There remains a need to develop drugs for treating these diseases. Summary of the invention

[0006] It has been unexpectedly discovered in the present invention that compounds, such as beauvericin (designated herein as GYT-088), are effective in treating eye diseases.

[0007] In one aspect, the present invention provides a method for treating an eye disease, the method comprising administering to a subject in need thereof a composition comprising a therapeutically effective amount of beauvericin having the following structural formula:

[0008]

[0009] In another aspect, the present invention also provides a use of beauvericin in preparing a medicament for treating eye diseases.

[0010] In another aspect, the present invention provides a pharmaceutical composition for treating an eye disease, comprising a combination of one or more pharmaceutically acceptable carriers and a therapeutically effective amount of beauvericin.

[0011] In one embodiment, the eye disease is caused by ocular degeneration due to neovascularization.

[0012] In some embodiments of the present invention, the eye disease is age-related macular degeneration (AMD), diabetic retinopathy (DR), or macular edema (ME). In a preferred embodiment of the present invention, the eye disease is non-proliferative diabetic retinopathy (NPDR) or wet age-related macular degeneration (wet AMD).

[0013] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The foregoing summary of the invention as well as the following detailed description of the invention will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the invention, presently preferred embodiments are shown in the accompanying drawings.

[0015] In the attached figure:

[0016] Figure 1 The images provided show angiogenesis in Matrigel plugs in groups treated with beauvericin (GYT-088, 2.4 mg / kg ip) and sorafenib (30 mg / kg ip), respectively.

[0017] Figure 2 A comparison of the heme content of each group is provided, showing that beauvericin provides a higher efficacy in inhibiting VEGF-induced angiogenesis.

[0018] Figure 3 It is shown that after oral administration of GYT-008, the test drug beauvericin (GYT-088) according to the present invention was observed in the eyes of animals, indicating that GYT-088 can cross the blood-brain barrier (BBB), especially the blood-retinal barrier (BRB), and enter the extravascular space of the retina and vitreous of the animals.

[0019] Figure 4 The images provided show the measurements of the fovea and macula center of this animal. DETAILED DESCRIPTION

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0021] In the present invention, it is firstly discovered that some compounds have significant effects in treating eye diseases such as age-related macular degeneration (AMD), diabetic retinopathy (DR), or macular edema (ME).

[0022] The present invention provides a method for treating eye diseases, comprising administering a composition comprising a therapeutically effective amount of beauvericin to an individual in need thereof, wherein the beauvericin is a cyclohexapeptide formed by coupling three LN-methylphenylanilines with three D-2-hydroxyisovaleric acids and has the following structural formula:

[0023]

[0024] The compounds of the present invention, such as beauvericin, can be synthesized or isolated from fungi such as Cordyceps cicadae, Cordyceps sobolifera, Cordyceps cicaicola, and Isaria fumosorosea according to conventional techniques or methods.

[0025] In the present invention, the compound is demonstrated to be effective in treating eye diseases by inhibiting VEGF-induced angiogenesis.

[0026] In one embodiment, the eye disease is caused by ocular degeneration due to neovascularization. Some examples of the eye disease include age-related macular degeneration (AMD), diabetic retinopathy (DR), and macular edema (ME). In a preferred embodiment of the present invention, the eye disease is non-proliferative diabetic retinopathy (NPDR) or wet age-related macular degeneration (wet AMD).

[0027] As used herein, the term "therapeutically effective amount" refers to an amount of a compound or agent that results in the treatment, cure, prevention, or improvement of a disease, condition, or side effect, or a reduction in the rate of progression of a disease or condition, compared to a corresponding individual not receiving the amount. The term also includes within its scope an amount that is effective in enhancing normal physiological function.

[0028] For use in treatment, the therapeutically effective amount of the compound is formulated into a pharmaceutical composition for administration. Therefore, the present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the compound beauvericin and one or more pharmaceutically acceptable carriers.

[0029] The term "pharmaceutically acceptable carrier" as used herein refers to an acceptable carrier, diluent or excipient in the sense of being compatible with the other ingredients of the formulation and harmless to the individual to whom the pharmaceutical composition is administered. Any carrier, diluent or excipient commonly known or used in the art can be used in the present invention, depending on the requirements of the pharmaceutical formulation.

[0030] According to the present invention, the pharmaceutical composition can be suitable for administration by any appropriate route, including but not limited to oral, rectal, nasal, topical, vaginal, or parenteral routes. In a particular embodiment of the present invention, the pharmaceutical composition is formulated for oral administration. Such preparations can be prepared by any method known in the pharmaceutical field.

[0031] The present invention is further illustrated by the following examples, which are provided by way of illustration and not limitation.

[0032] Example

[0033] Example 1

[0034] In vivo Matrigel plug angiogenesis assay

[0035] Matrigel plugs are made from a gelatinous mixture of proteins secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells, produced and sold by Corning LifeSciences and BD Biosciences. Matrigel resembles the complex extracellular environment found in many tissues and is used by cell biologists as a matrix (basement membrane matrix) for culture assays.

[0036] The experimental animals were 5-8 week old B57l / 6 mice. Matrigel was mixed with PBS at a ratio of 4:1 to obtain Matrigel plugs. Matrigel plugs containing 30 IU heparin and VEGF 150 ng / ml (500 ul / mouse) were administered to mice, and then divided into four groups (four mice per group), intraperitoneally injected or orally administered with beauvericin (GYT-088 group), sorafenib (sorafenib group) as a positive control, VEGF alone (vehicle group), and no treatment (control group). One day later, the animals were intraperitoneally injected with 30 mg / kg sorafenib, or intraperitoneally injected with 2.4 mg / kg beauvericin (0.1 cc / mouse / day), or orally administered with 24 mg / kg beauvericin, once a day for 6 days. On the 7th day, the animals were sacrificed and the Matrigel plugs were removed to observe angiogenesis. The hemoglobin of each mouse was measured using a Drabkin's reagent kit.

[0037] The results are as follows Figure 1 and Figure 2 As provided in Figure 1 As shown in the figure, in the group injected with beauvericin intraperitoneally (GYT-088 group) or the group with sorafenib (sorafenib group) as a positive control, VEGF-induced angiogenesis was inhibited. Figure 2 As shown, based on the measured heme content, beauvericin provided a higher efficacy in inhibiting VEGF-induced angiogenesis.

[0038] In conclusion, beauvericin provides angiogenesis-inhibiting efficacy that is even superior to sorafenib.

[0039] Example 2

[0040] To evaluate the efficacy of different doses of beauvericin in improving naturally occurring diabetic macular edema in rhesus monkeys for 60 days, and to provide a basis for clinical trial dosage design and safety studies.

[0041] Rhesus Macaques were used in the evaluation.

[0042] Primary endpoint:

[0043] Retinal thickness was measured by OCT to quantify retinal thickness before and after treatment.

[0044] Retinal thickness was assessed using optical coherence tomography (OCT) using two methods: (1) at the fovea and 1.5 mm to either side of the fovea (three-point method), and (2) using a 52-point grid to determine the thickness of the macula in different thickness groups (250-300, 301-350, 351-400 μm, etc.). This method was developed by the Primed team in collaboration with Dr. Timothy Kern, a renowned diabetic retinopathy expert at Case Western Reserve University. See the ARVO 2018 poster: Assessment of macular thickening in spontaneously diabetic rhesus monkeys.

[0045] Secondary endpoints and safety parameters:

[0046] (1) Fundus photography (FP). Detect changes in bleeding, exudate, or vascular lesions before and after treatment; and

[0047] (2) Effects of drugs on metabolism, fat profile, liver and kidney function.

[0048] Non-human primates (NHP) are very similar to humans in terms of physiological functions and eye structure. PriMed has a unique large population of NHPs (n=about 1500) with spontaneous metabolic diseases and 5 years of historical clinical / metabolic data, including fasting plasma glucose (FPG), insulin resistance, body mass index (BMI), triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), and high-density lipoprotein cholesterol (HDL-c). Color Doppler ultrasound, Kowa fundus camera, Heidelberg optical coherence tomography (Heidelberg OCT), slit lamp, and ophthalmoscope are used to diagnose the diabetic state in non-human primates (NHPs), showing that the disease characteristics of spontaneous diabetic macular edema (DME) in rhesus macaques are very similar to those in clinical patients, showing vascular leakage, retinal thickening, exudation, or hemorrhage. Rhesus macaques with spontaneous diabetic macular edema (DME) provide an ideal model for human DME / diabetic retinopathy (DR) research and new drug development.

[0049] Inclusion Criteria

[0050] (1) Female, 13-18 years old (equivalent to 40-60 years old human adults), weighing 6-7 kg;

[0051] (2) Diabetes duration: more than 3 years;

[0052] (3) diabetic macular edema (DME) in one or both eyes;

[0053] (4) Diabetic macular edema (DME): retinal thickness measured by Heidelberg Spectralis OCT and diagnosed by fundus photography (FP) / fluorescein fundus angiography (FFA) with a Kowa VX-20 fundus camera in the last six months;

[0054] (5) Central Foveal Thickness (CRT) ≥ 260 μm and / or Early Treatment Diabetic Retinopathy Study (ETDRS) area average thickness ≥ 319 μm. (Retinal thickening was defined by the mean and standard deviations (SD) of CRT and ETDRS thickness in normal macaques). Retinal thickening was defined as thickness exceeding 2 standard deviations of the normal mean. Non-diabetic macaques: The mean foveal thickness of the retina of non-diabetic animals was 192 ± 11 μm, with values ​​of 307 ± 15 and 328 ± 13 measured at 1.5 mm temporal and nasal to the fovea, respectively. This method was developed by the Primed team in collaboration with Dr. Timothy Kern, a renowned diabetic retinopathy expert at Case Western Reserve University. Please see the ARVO 2018 poster: Assessment of macular thickening in spontaneously diabetic rhesus macaques; and

[0055] (6) Presence / absence of vascular leakage or non-perfused areas.

[0056] Exclusion criteria

[0057] (1) Hypertension stage 3 or above;

[0058] (2) Progressive ocular or systemic inflammation or infection; intraocular hypertension: intraocular pressure (IOP)>21 mmHg;

[0059] (3) Laboratory tests showed significant impairment of kidney or liver function;

[0060] (4) Any other medical history that may affect the efficacy of the treatment.

[0061] Group size

[0062] Six rhesus macaques with spontaneous diabetic macular edema (DME) (with medical history within one year) will be provided, and four of the six monkeys will undergo fundus angiography (FA), fundus photography (FP), optical coherence tomography (OCT), and slit lamp examinations, and blood biochemical tests in the early stage of the study (4 weeks) before treatment. The optical coherence tomography (OCT) will divide the four monkeys into two groups based on retinal thickness:

[0063] (1) Group 1, vehicle group, N = 1;

[0064] (2) Group 2, experimental group, N = 3;

[0065] (Route of administration: oral, once a day; Dose: total basal dose (TBD)).

[0066] Outcome measures:

[0067] Main efficacy parameters:

[0068] Retinal thickness includes central subfield thickness (CST) and ETDRS thickness. Retinal thickness was measured before administration and once on the 14th day (D14), 30th day (D30), and 60th day (D60) after administration. The results of D14 and D30 were used to determine whether to add the test on the 45th day (D45).

[0069] Secondary efficacy and safety parameters:

[0070] (1) Fundus photography: The baseline was before administration, and the 30th day (D30) and 60th day (D60) after administration were taken, for a total of 3 times;

[0071] (2) Fundus fluorescein angiography (FFA): the baseline was the time before drug administration;

[0072] (3) Intraocular pressure and blood pressure measurement: once before and once after medication, a total of 2 times;

[0073] (4) Evaluation of clinical signs of the eyes by daily cage-side observation;

[0074] (5) Determination of metabolic parameters and liver and kidney function: FPG, FRA, LDL, HDL, TC, TG: before administration and once on the 14th day (D14), 30th day (D30), 45th day (D45), and 60th day (D60) after administration;

[0075] (6) Slit lamp: once before medication and once after medication, a total of 2 times;

[0076] (7) Body weight: before administration and once on the 30th day (D30) and 60th day (D60) after administration;

[0077] (8) Food intake / clinical observations: including abnormal signs, symptoms, and behaviors, once a day.

[0078] Fundus photography (FP) and fundus fluorescein angiography (FFA)

[0079] (1) Frequency: Fluorescein fundus angiography (FFA) was performed once before administration.

[0080] (2) Instrument: KOWA VX-20.

[0081] (3) Purpose: To examine retinal vascular damage and leakage.

[0082] (4)FFA image acquisition procedure:

[0083] Prior to acquiring images, animals were anesthetized by intramuscular injection of a ketamine:xylazine mixture (1:1, 8 mg / kg ketamine). After anesthesia, 2 drops of Tropicamide phenylephrine eye drops were applied to each eye to dilate the pupil. After anesthesia, the animal was placed in a dark room until the pupil diameter was greater than 6 mm. A self-retaining eyelid spectroscope was placed in the eye. The fundus of the dominant eye was well focused at the posterior pole.

[0084] (5) The timer was started when 10% sodium fluorescein (Alcon Laboratories, USA) was injected via the femoral vein at a dose of 0.075 mL / kg. A series of posterior pole photographs of the dominant eye were taken every second for the first 30 seconds and every 2-3 seconds for the next 30 seconds. Both eyes were photographed at 5 minutes and 10-15 minutes.

[0085] result

[0086] Vehicle group: #6189 showed mild non-proliferative diabetic retinopathy (NPDR) before administration. As shown in Table 1, no significant changes were found before and after administration by fundus photography (FP) analysis. GYT-088 group: #251, #2003, and #3536 showed mild or moderate non-proliferative diabetic retinopathy (NPDR). Fundus photography (FP) analysis improved the characteristics of non-proliferative diabetic retinopathy (NPDR) in #2003 on day 28 (D28) after administration. No significant changes were found before and after administration of other animals by fundus photography (FP) analysis at any time point, please refer to Table 1.

[0087] Table 1 Non-proliferative diabetic retinopathy (NPDR) related changes after treatment

[0088]

[0089]

[0090] Results are available on Figure 4On the 14th day after administration, it was found in the experimental animal #2003 (NPDR group) that the central macular thickness (CMT) changed from 204μm and 210μm in the left and right eyes to 197μm and 202μm, respectively, a decrease of 7μm and 8μm; the thickness of the fovea changed from 257μm and 260μm in the left and right eyes to 249μm and 251μm, respectively, a decrease of 8μm and 9μm. On the other hand, it was found that in the animal #6189 (vehicle group) administered with the vehicle, the change in central macular thickness or foveal thickness was within the range of ±2μm, as normally found in normal animals. On the 56th day after administration, the central macular thickness decreased by 6μm and 8μm, and the macular thickness of the left and right eyes decreased by 3μm and 5μm, respectively. Therefore, it can be concluded that GYT-088 provides efficacy in diabetic macular edema.

[0091] Table 2 Changes in thickness of diabetic macular edema (DME)

[0092]

[0093]

[0094] Although this specification contains many details, these details should not be interpreted as limiting the scope of the invention or the scope of protection that can be claimed, but should be understood as a description of the features specific to a particular embodiment or example of the invention. Certain features described in this specification in the context of separate embodiments or examples may also be implemented in combination in a single embodiment.

Claims

1. Use of beauvericin in the preparation of a medicament for treating eye diseases, The eye disease is diabetic retinopathy (DR) or macular edema (ME), Beauvericin has the following structure: 。 2. The use as claimed in claim 1, wherein the eye disease is caused by ocular degeneration due to neovascularization.

3. The use according to claim 1, wherein the diabetic retinopathy (DR) is non-proliferative diabetic retinopathy (NPDR).

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