Preparation composition for preventing and treating eye diseases and delaying visual decline and preparation method thereof

Abstract

The invention discloses a preparation composition for preventing and treating eye diseases and delaying visual decline and a preparation method thereof. The composition is prepared from fructus lycii, semen cassiae, blueberries, marigold, eyebright, ligusticum wallichii, pseudo-ginseng, roxburgh rose, pleurotus citrinopileatus, bilberry, haematococcus pluvialis, xanthophyll, zeaxanthin, retinol, ergothioneine, beta-carotene, vitamin E, organic selenium, phospholipid DHA, procyanidine, curcumin and lycopene. The composition can prevent and treat eye diseases and delay visual decline. The traditional Chinese medicine composition can relieve asthenopia, can relieve the symptoms of blurred vision, dry eyes, soreness and swelling, pain and the like, and can treat macular degeneration. The composition has no side effect on a human body and is high in safety; injection administration and operation are avoided, no sequelae are left, and the medicine is convenient to take and carry by a patient; the preparation process is simple, the performance is stable and reliable, the production efficiency is high, the cost is low, and the application prospect is wide. The invention further discloses a polypeptide for relieving asthenopia, wherein the sequence of the polypeptide is shown as SEQ ID NO: 1.

Description

[0001] This application is a divisional application of the patent application with the application date of December 16, 2024, application number 202411849556.8, and invention-creation name of "Preparation Composition for Preventing and Treating Eye Diseases and Delaying Vision Decline and Its Preparation Method". Technical Field

[0002] The present invention belongs to the field of biomedicine, and particularly relates to polypeptides, compositions, preparations for preventing and treating eye diseases and delaying vision decline, and their preparation methods. Background Art

[0003] Visual fatigue, also known as eye fatigue or ocular fatigue, refers to the symptoms of fatigue and discomfort in the eyes caused by prolonged use or excessive load of the eyes. This condition is common in prolonged close-range eye use, such as prolonged use of electronic devices (mobile phones, computers, tablets), reading, driving, etc. Common symptoms of visual fatigue include but are not limited to: eye soreness or dryness, blurred vision or temporary vision decline, a burning or foreign body sensation in the eyes, headache or discomfort in the neck and shoulders, eye congestion, tearing or frequent blinking, and light sensitivity.

[0004] The main causes of visual fatigue include but are not limited to: prolonged use of electronic screens; poor lighting; prolonged close-range eye use; bad eye use habits; uncorrected refractive errors of the eyes (myopia, hyperopia, astigmatism), or inappropriate glasses.

[0005] Methods for relieving visual fatigue include but are not limited to: following the "20-20-20" rule, looking at an object 20 feet (about 6 meters) away for at least 20 seconds every 20 minutes of work; maintaining good eye use habits, adjusting the screen brightness, maintaining a proper eye use distance and posture, and ensuring moderate lighting in the working environment; taking regular breaks and relaxing the eyes, closing the eyes, eye massage or hot compress; keeping the air humid; using artificial tears. However, the effects of these methods are still limited.

[0006] Age-related Macular Degeneration (AMD) is an age-related eye disease that mainly affects the macula area of the retina, resulting in a gradual loss of central vision. This is one of the main causes of blindness in the elderly. The macula is the area of the retina responsible for clear central vision, and central vision is crucial for daily activities such as reading, driving, and recognizing faces. Age-related Macular Degeneration is divided into dry Age-related Macular Degeneration (the most common, accounting for most cases) and wet Age-related Macular Degeneration (less common but more severe and faster-progressing than dry). Due to abnormal new blood vessels growing and leaking fluid or blood under the retina, the macula is damaged. It often leads to rapid vision deterioration, which may occur within a few months or even weeks.

[0007] Common symptoms of macular degeneration include, but are not limited to: blurred central vision, difficulty seeing details clearly, especially when reading or looking at distant objects; distorted vision, where straight lines appear bent or twisted; central scotoma, where a shadow or blind spot may appear in the center of the visual field; color vision changes, difficulty distinguishing colors or reduced color contrast.

[0008] Major risk factors for macular degeneration include, but are not limited to: age, with a higher incidence in people over 50 years old; family history, people with a family history of macular degeneration are at higher risk; obesity and cardiovascular diseases, people who are overweight or have cardiovascular diseases also have a higher risk; long-term ultraviolet exposure, excessive sunlight exposure may increase the risk of macular degeneration.

[0009] Drug treatments for wet macular degeneration include, but are not limited to: photodynamic therapy, which combines drugs and laser treatment to treat neovascularization in wet macular degeneration; laser treatment, for some patients with wet macular degeneration, abnormal blood vessels can be destroyed by laser to prevent further vision damage. However, the above methods are inconvenient and have limited effects.

[0010] Clinically, there is an urgent need for drugs to relieve visual fatigue and treat macular degeneration. Summary of the Invention

[0011] In one aspect of the present invention, there is provided a composition, characterized in that the composition comprises the following components in parts by weight:

[0012] Lycium barbarum 25 - 110 parts by weight, Cassia obtusifolia 25 - 95 parts by weight, blueberry 10 - 90 parts by weight, Tagetes erecta 10 - 95 parts by weight, Euphrasia officinalis 10 - 90 parts by weight, Ligusticum wallichii 10 - 90 parts by weight, Panax notoginseng 10 - 90 parts by weight, Rosa roxburghii Tratt 20 - 90 parts by weight, Pleurotus citrinopileatus 15 - 90 parts by weight, Vaccinium vitis-idaea 20 - 90 parts by weight, Haematococcus pluvialis 20 - 90 parts by weight, lutein 10 - 85 parts by weight, zeaxanthin 10 - 90 parts by weight, retinol 10 - 90 parts by weight, ergothioneine 20 - 95 parts by weight, β-carotene 25 - 100 parts by weight, vitamin E 20 - 95 parts by weight, organic selenium 10 - 90 parts by weight, phospholipid DHA 20 - 90 parts by weight, proanthocyanidins 15 - 90 parts by weight, curcumin 15 - 80 parts by weight, lycopene 15 - 95 parts by weight.

[0013] In some embodiments of the present invention, the composition comprises the following components in parts by weight:

[0014] 25 - 110 parts by weight of wolfberry, 25 - 95 parts by weight of cassia seed, 10 - 90 parts by weight of blueberry, 10 - 95 parts by weight of marigold, 10 - 90 parts by weight of eyebright, 10 - 90 parts by weight of chuanxiong, 10 - 90 parts by weight of notoginseng, 20 - 90 parts by weight of roxburgh rose, 15 - 90 parts by weight of Pleurotus citrinopileatus, 20 - 90 parts by weight of bilberry, 20 - 90 parts by weight of Haematococcus pluvialis, 10 - 85 parts by weight of lutein, 10 - 90 parts by weight of zeaxanthin, 10 - 90 parts by weight of retinol, 20 - 95 parts by weight of ergothioneine, 25 - 100 parts by weight of β - carotene, 20 - 95 parts by weight of vitamin E, 10 - 90 parts by weight of selenyl - L - cysteine, 20 - 90 parts by weight of phospholipid DHA, 15 - 90 parts by weight of procyanidin, 15 - 80 parts by weight of curcumin, 15 - 95 parts by weight of lycopene.

[0015] In some embodiments of the present invention, the composition comprises the following components in parts by weight:

[0016] 80 parts by weight of wolfberry, 80 parts by weight of cassia seed, 40 parts by weight of blueberry, 70 parts by weight of marigold, 80 parts by weight of eyebright, 50 parts by weight of chuanxiong, 40 parts by weight of notoginseng, 30 parts by weight of roxburgh rose, 80 parts by weight of Pleurotus citrinopileatus, 30 parts by weight of bilberry, 80 parts by weight of Haematococcus pluvialis, 70 parts by weight of lutein, 80 parts by weight of zeaxanthin, 30 parts by weight of retinol, 50 parts by weight of ergothioneine, 30 parts by weight of β - carotene, 30 parts by weight of vitamin E, 40 parts by weight of selenyl - L - cysteine, 30 parts by weight of phospholipid DHA, 80 parts by weight of procyanidin, 30 parts by weight of curcumin, 30 parts by weight of lycopene.

[0017] In some embodiments of the present invention, the wolfberry is the dried fruit of wolfberry. In some embodiments of the present invention, the cassia seed is the dried ripe seed of cassia. In some embodiments of the present invention, the blueberry is the dried fruit of blueberry. In some embodiments of the present invention, the marigold is the dried leaf. In some embodiments of the present invention, the eyebright is the dried whole herb. In some embodiments of the present invention, the chuanxiong is the dried rhizome. In some embodiments of the present invention, the notoginseng is the dried rhizome. In some embodiments of the present invention, the roxburgh rose is the dried fruit. In some embodiments of the present invention, the Pleurotus citrinopileatus is the dried fruiting body. In some embodiments of the present invention, the bilberry is the dried fruit.

[0018] In some embodiments of the present invention, the Haematococcus pluvialis is Haematococcus pluvialis powder. In some embodiments of the present invention, the Haematococcus pluvialis powder is purchased from ZOLANBIO. In some embodiments of the present invention, the lutein is purchased from Shaanxi Yikanglong Biotechnology. In some embodiments of the present invention, the zeaxanthin is purchased from Wuhan Fengtai Weiyuan Technology. In some embodiments of the present invention, the retinol is purchased from Jiangxi Ruiweier Biotechnology. In some embodiments of the present invention, the ergothioneine is purchased from Shenzhen Xijia Biotechnology. In some embodiments of the present invention, the β-carotene is purchased from Sigma. In some embodiments of the present invention, the vitamin E is purchased from Hebei Zhenjia New Materials. In some embodiments of the present invention, the organic selenium is selenocysteine. In some embodiments of the present invention, the organic selenium is seleno-L-cysteine. In some embodiments of the present invention, the seleno-L-cysteine is purchased from Wuhan Innovo Pharmaceutical Technology. In some embodiments of the present invention, the CAS number of the phospholipid DHA is 59403-52-0. In some embodiments of the present invention, the phospholipid DHA is purchased from Product catalog number HY-W440983. In some embodiments of the present invention, the proanthocyanidins are purchased from Proanthocyanidins, catalog number HY-N0794. In some embodiments of the present invention, the curcumin is purchased from Shaanxi Yikanglong Biotechnology. In some embodiments of the present invention, the lycopene is purchased from Nanjing Daosifu Biotechnology.

[0019] In some embodiments of the present invention, the composition is the composition AMX-EYE-01, AMX-EYE-02 or AMX-EYE-03 described in Example 1. In some embodiments of the present invention, the composition is the composition AMX-EYE-03 described in Example 1. In some embodiments of the present invention, the composition contains 0.01%-0.03% by mass of the polypeptide AMX-EYE-P1. The amino acid sequence of the polypeptide AMX-EYE-P1 is shown in SEQ ID NO:1.

[0020] In some embodiments of the present invention, the composition is prepared by the method described in Example 1.

[0021] In some embodiments of the present invention, the composition is prepared by the following method:

[0022] Take 80 grams of wolfberry, 80 grams of cassia seed, 40 grams of blueberry, 70 grams of calendula, 80 grams of eyebright, 50 grams of chuanxiong, 40 grams of notoginseng, 30 grams of rosa roxburghii tratt, 80 grams of pleurotus citrinopileatus, 30 grams of bilberry; wash them separately with water, air-dry at room temperature, and crush them to an average particle size of 500 microns; then mix them evenly with 80 grams of haematococcus pluvialis, 70 grams of lutein, 80 grams of zeaxanthin, 30 grams of retinol, 50 grams of ergothioneine, 30 grams of β-carotene, 30 grams of vitamin E, 40 grams of seleno-L-cysteine, 30 grams of phosphatidyl DHA, 80 grams of proanthocyanidins, 30 grams of curcumin, and 30 grams of lycopene to obtain the product.

[0023] In another aspect of the present invention, there is provided a method for preparing a composition, which is characterized by comprising the following steps:

[0024] Take 25 - 110 parts by weight of wolfberry, 25 - 95 parts by weight of cassia seed, 10 - 90 parts by weight of blueberry, 10 - 95 parts by weight of calendula, 10 - 90 parts by weight of eyebright, 10 - 90 parts by weight of chuanxiong, 10 - 90 parts by weight of notoginseng, 20 - 90 parts by weight of rosa roxburghii tratt, 15 - 90 parts by weight of pleurotus citrinopileatus, 20 - 90 parts by weight of bilberry, 20 - 90 parts by weight of haematococcus pluvialis, 10 - 85 parts by weight of lutein, 10 - 90 parts by weight of zeaxanthin, 10 - 90 parts by weight of retinol, 20 - 95 parts by weight of ergothioneine, 25 - 100 parts by weight of β-carotene, 20 - 95 parts by weight of vitamin E, 10 - 90 parts by weight of seleno-L-cysteine, 20 - 90 parts by weight of phosphatidyl DHA, 15 - 90 parts by weight of proanthocyanidins, 15 - 80 parts by weight of curcumin, 15 - 95 parts by weight of lycopene, mix them evenly to obtain the product.

[0025] In some embodiments of the present invention, the preparation method comprises the following steps: Take 80 grams of wolfberry, 80 grams of cassia seed, 40 grams of blueberry, 70 grams of calendula, 80 grams of eyebright, 50 grams of chuanxiong, 40 grams of notoginseng, 30 grams of rosa roxburghii tratt, 80 grams of pleurotus citrinopileatus, 30 grams of bilberry; wash them separately with water, air-dry at room temperature, and crush them to an average particle size of 500 microns; then mix them evenly with 80 grams of haematococcus pluvialis, 70 grams of lutein, 80 grams of zeaxanthin, 30 grams of retinol, 50 grams of ergothioneine, 30 grams of β-carotene, 30 grams of vitamin E, 40 grams of seleno-L-cysteine, 30 grams of phosphatidyl DHA, 80 grams of proanthocyanidins, 30 grams of curcumin, and 30 grams of lycopene to obtain the product.

[0026] In some specific embodiments of the present invention, the preparation method of the composition is as described in Example 1.

[0027] In some embodiments of the present invention, the preparation method of the composition comprises:

[0028] The aforementioned composition is irradiated for sterilization, granulated, dried, and sized to obtain a granule agent.

[0029] In some embodiments of the present invention, the preparation method of the composition comprises:

[0030] The aforementioned composition is irradiated for sterilization, granulated, dried, and sized to obtain granules; the granules are mixed with pharmaceutically acceptable excipients and then filled into a capsule shell, or the granules are directly filled into a capsule shell to obtain a capsule.

[0031] In some embodiments of the present invention, the granulation method is to add ethanol with a volume fraction of 65%-75% and a weight of 3%-6% of the sterilized mixture to make a soft material, and then granulate through a 10-mesh to 16-mesh sieve.

[0032] In some embodiments of the present invention, the drying temperature is 50°C - 70°C and the time is 2 - 4 hours.

[0033] In some embodiments of the present invention, the sterilization is irradiation sterilization, and the environment for the irradiation sterilization is: cobalt-60 radiation sterilization with an irradiation dose ≤ 6 kGy.

[0034] In some embodiments of the present invention, the preparation method of the composition comprises: irradiating the aforementioned composition for sterilization with 60Co radiation at an irradiation dose of 3 kGy; adding ethanol with a concentration of 70% (V / V) and a weight of 4% of the sterilized mixture to make a soft material, and then granulating through a 12-mesh sieve; drying at 55°C for 3 hours; sizing to obtain a granule agent.

[0035] In some embodiments of the present invention, the preparation method of the composition comprises: irradiating the aforementioned composition for sterilization with 60Co radiation at an irradiation dose of 3 kGy; adding ethanol with a concentration of 70% (V / V) and a weight of 4% of the sterilized mixture to make a soft material, and then granulating through a 12-mesh sieve; drying at 55°C for 3 hours; sizing to obtain composition granules; filling the composition granules into a capsule shell to obtain a capsule.

[0036] In some embodiments of the present invention, each capsule shell is filled with 400 - 800 mg of the composition granules.

[0037] In some embodiments of the present invention, each capsule shell is filled with 500 mg, 600 mg, or 750 mg of the composition granules.

[0038] In some embodiments of the present invention, the capsule shell is a hard capsule shell and the capsule is a hard capsule.

[0039] In some embodiments of the present invention, the capsule shell is a soft capsule shell and the capsule is a soft capsule.

[0040] In another aspect of the present invention, there is provided a polypeptide, characterized in that the polypeptide comprises the amino acid sequence shown in SEQ ID NO: 1 or 2.

[0041] In some embodiments of the present invention, the amino acid sequence of the polypeptide is as shown in SEQ ID NO: 1 or 2.

[0042] In some embodiments of the present invention, the amino acid sequence of the polypeptide is as shown in SEQ ID NO: 1.

[0043] In another aspect of the present invention, there is provided a preparation, characterized in that the preparation comprises the composition as described in the present invention.

[0044] In some specific embodiments of the present invention, the preparation is a pharmaceutical preparation.

[0045] In some specific embodiments of the present invention, the preparation is an oral dosage form.

[0046] In some specific embodiments of the present invention, the dosage form of the preparation is: capsule, granule, tablet, oral liquid or pill.

[0047] In some embodiments of the present invention, the preparation is optionally selected from: hard capsule, soft capsule, granule, tablet.

[0048] In some embodiments of the present invention, the composition described in any one of the present invention is irradiated for sterilization, granulated, dried, sized, and a granule is obtained.

[0049] In some embodiments of the present invention, the composition described in any one of the present invention is irradiated for sterilization, granulated, dried, sized, and a granule is obtained; the granule is filled into a capsule shell to obtain a capsule.

[0050] In some embodiments of the present invention, the composition described in any one of the present invention is irradiated for sterilization, granulated, dried, sized, and a granule is obtained; the granule is filled into a soft capsule shell to obtain a soft capsule.

[0051] In some embodiments of the present invention, the composition described in any one of the present invention is irradiated for sterilization, auxiliary materials are added, and tablets are prepared by tabletting.

[0052] In some embodiments of the present invention, the preparation method of the preparation comprises:

[0053] The foregoing composition is irradiated for sterilization, granulated, dried, sized, and a granule is obtained; the granule is mixed with pharmaceutically acceptable auxiliary materials and then filled into a capsule shell or directly filled into a capsule shell to obtain a capsule.

[0054] In some embodiments of the present invention, the granulation method is to add ethanol with a volume fraction of 65%-75% and a weight of 3%-6% of the mixture after sterilization to make soft materials, and then granulate through a 10-mesh to 16-mesh sieve.

[0055] In some embodiments of the present invention, the drying temperature is 50°C - 70°C, and the time is 2 - 4 hours.

[0056] In some embodiments of the present invention, the sterilization is irradiation sterilization, and the environment of the irradiation sterilization is: cobalt-60 radiation sterilization, and the irradiation dose ≤ 6 kGy.

[0057] In some embodiments of the present invention, the preparation method of the preparation includes: irradiating and sterilizing the aforementioned composition with 60Co radiation, with an irradiation dose of 3 kGy; adding ethanol with a concentration of 70% (V / V) and a weight of 4% of the mixture after sterilization to make soft materials, and then granulating through a 12-mesh sieve; drying at 55°C for 3 hours; sizing to obtain granules.

[0058] In some embodiments of the present invention, the preparation method of the preparation includes: irradiating and sterilizing the aforementioned composition with 60Co radiation, with an irradiation dose of 3 kGy; adding ethanol with a concentration of 70% (V / V) and a weight of 4% of the mixture after sterilization to make soft materials, and then granulating through a 12-mesh sieve; drying at 55°C for 3 hours; sizing to obtain composition granules; filling the composition granules into a capsule shell to obtain capsules.

[0059] In some embodiments of the present invention, each capsule shell is filled with 400 - 800 mg of the aforementioned composition granules.

[0060] In some embodiments of the present invention, each capsule shell is filled with 500 mg, 600 mg or 750 mg of the aforementioned composition granules.

[0061] In some embodiments of the present invention, the capsule shell is a hard capsule shell, and the capsule is a hard capsule.

[0062] In some embodiments of the present invention, the capsule shell is a soft capsule shell, and the capsule is a soft capsule.

[0063] In some embodiments of the present invention, the preparation is any one of the preparations described in Example VII.

[0064] In another aspect of the present invention, there is provided the use of the composition as described in the present invention, the preparation as described in the present invention, and the polypeptide as described in the present invention in the preparation of a drug for relieving eye fatigue or treating macular degeneration.

[0065] In some embodiments of the present invention, there is provided the use of the composition as described in the present invention, the preparation as described in the present invention, and the polypeptide as described in the present invention in the preparation of a medicament for relieving asthenopia or treating macular degeneration.

[0066] In some embodiments of the present invention, there is provided the use of the composition as described in the present invention, the preparation as described in the present invention, and the polypeptide as described in the present invention in the preparation of a medicament for reducing the thickness of Bruch's membrane.

[0067] In some embodiments of the present invention, there is provided the use of the composition as described in the present invention, the preparation as described in the present invention, and the polypeptide as described in the present invention in the preparation of a medicament for preventing or treating macular degeneration by reducing the thickness of Bruch's membrane.

[0068] In some embodiments of the present invention, there is provided the use of the composition as described in the present invention, the preparation as described in the present invention, and the polypeptide as described in the present invention in the preparation of a medicament for treating macular degeneration.

[0069] In some embodiments of the present invention, there is provided the use of the composition as described in the present invention in the preparation of a medicament for relieving asthenopia.

[0070] In some embodiments of the present invention, there is provided the use of the polypeptide as described in the present invention in the preparation of a medicament for relieving asthenopia.

[0071] In some embodiments of the present invention, there is provided the use of the composition as described in the present invention in the preparation of a medicament for treating macular degeneration.

[0072] In some embodiments of the present invention, there is provided the use of the polypeptide as described in the present invention in the preparation of a medicament for treating macular degeneration.

[0073] Technical effects of the technical solution of the present invention:

[0074] The composition, preparation, and polypeptide of the present invention can prevent and treat eye diseases and delay vision decline. Specifically, it can relieve asthenopia, reduce symptoms such as blurred vision, dry eyes, soreness, and pain, and can treat macular degeneration. It can prevent and treat vision problems such as vision decline, night blindness, floaters, glaucoma, and cataract caused by macular lesions, retinitis pigmentosa, and diabetic retinopathy, and can improve vision to the greatest extent.

[0075] The composition of the present invention rationally proportions each component, has no side effects on the human body, high safety, and remarkable curative effect. The composition described in the present invention avoids injection administration and surgery, leaves no sequelae, and is convenient for patients to take and carry. The preparation process of the preparation is simple, the performance is stable and reliable, the production efficiency is high, the production cost is low, and it has broad application prospects. Detailed implementation manners

[0076] The present invention will be further described below by way of examples, but the present invention is not limited to the scope of the described examples. For the experimental methods without specific conditions noted in the following examples, they are carried out according to conventional methods and conditions, or selected according to the product specifications. Unless otherwise specified, the reagents and raw materials used in the present invention are all commercially available.

[0077] Example 1:

[0078] Take 80 grams of wolfberry (dry fruit of wolfberry), 80 grams of cassia seed (dry mature seed of cassia), 40 grams of blueberry (dry fruit), 70 grams of marigold (dry leaf), 80 grams of eyebright (dry whole herb), 50 grams of rhizoma chuanxiong (dry rhizome), 40 grams of notoginseng (dry rhizome), 30 grams of rosa roxburghii tratt (dry fruit), 80 grams of pleurotus citrinopileatus (dry fruiting body), 30 grams of bilberry (dry fruit); wash them separately with water, air-dry at room temperature, and crush them to an average particle size of 500 microns; mix them evenly to obtain a solid powder composition AMX-EYE-01.

[0079] Take 80 grams of haematococcus pluvialis powder (purchased from ZOLANBIO), 70 grams of lutein (purchased from Shaanxi Yikanglong Biotechnology), 80 grams of zeaxanthin (purchased from Wuhan Fengtai Weiyuan Technology), 30 grams of retinol (purchased from Jiangxi Ruiweier Biotechnology), 50 grams of ergothioneine (purchased from Shenzhen Xijia Biotechnology), 30 grams of β-carotene (Sigma), 30 grams of vitamin E (purchased from Hebei Zhenjia New Materials), 40 grams of seleno-L-cysteine (purchased from Wuhan Innovo Pharmaceutical Technology), 30 grams of phospholipid DHA (CAS No. 59403-52-0, product catalog number HY-W440983), 30 grams of procyanidin ( procyanidin, catalog number HY-N0794), 80 grams of curcumin (purchased from Shaanxi Yikanglong Biotechnology), 30 grams of lycopene (purchased from Nanjing Daosifu Biotechnology); mix them evenly to obtain a solid powder composition AMX-EYE-02.

[0080] Take 80 grams of wolfberries (dried fruits of Lycium barbarum), 80 grams of cassia seeds (dried ripe seeds of Cassia obtusifolia), 40 grams of blueberries (dried fruits), 70 grams of marigolds (dried leaves), 80 grams of eyebrights (dried whole herbs), 50 grams of chuanxiong rhizomes (dried rhizomes), 40 grams of notoginseng (dried rhizomes), 30 grams of roxburgh roses (dried fruits), 80 grams of Pleurotus citrinopileatus (dried fruiting bodies), 30 grams of bilberries (dried fruits); wash them separately with water, air-dry at room temperature, and crush them to an average particle size of 500 microns; then mix them evenly with 80 grams of Haematococcus pluvialis powder (purchased from ZOLANBIO), 70 grams of lutein (purchased from Shaanxi Yikanglong Biotechnology), 80 grams of zeaxanthin (purchased from Wuhan Fengtai Weiyuan Technology), 30 grams of retinol (purchased from Jiangxi Ruiweier Biotechnology), 50 grams of ergothioneine (purchased from Shenzhen Xijia Biotechnology), 30 grams of β-carotene (Sigma), 30 grams of vitamin E (purchased from Hebei Zhenjia New Materials), 40 grams of seleno-L-cysteine (purchased from Wuhan Yinnuo Pharmaceutical Technology), 30 grams of phosphatidyl DHA (CAS No. 59403-52-0, product catalog number HY-W440983) 30 grams, proanthocyanidins ( Proanthocyanidins, catalog number HY-N0794) 80 grams, curcumin (purchased from Shaanxi Yikanglong Biotechnology) 30 grams, lycopene (purchased from Nanjing Daosifu Biotechnology) 30 grams, and mix them evenly to obtain a solid powder composition AMX-EYE-03.

[0081] Example 2: Test for the function of relieving visual fatigue

[0082] Take healthy 8-month-old New Zealand rabbits (weight 4.45 - 4.51 kg), with an equal number of males and females. Randomly divide them into groups of 10 each. The experimental group animals were intragastrically administered a suspension of composition AMX-EYE-01, AMX-EYE-02, or AMX-EYE-03 prepared by the method described in Example 1 (prepared by stirring 300 mg of the composition in 5 mL of water for 5 min) every day from the 1st to the 45th day. The negative control group was replaced with an equal volume of normal saline every day. From the 31st to the 34th day, all animals were irradiated with light at an intensity of 5000 Lux every day. Starting from the 35th day, the congestion condition of the rabbit eyes was observed and recorded every day, and the recovery time of iris and conjunctival congestion (the number of days from the 35th day to the day when the congestion recovered) was calculated. The results are shown in Table 1 (the average value of each group). Compared with other groups, the AMX-EYE-03 group can significantly reduce the recovery time of iris congestion and conjunctival congestion, and both have statistically significant differences (P < 0.01). It shows that the composition AMX-EYE-03 can significantly relieve eye fatigue, and there is a synergistic effect between the components, achieving an unexpected technical effect.

[0083] Table 1. Test for the function of relieving visual fatigue

[0084]

[0085] Example 3: Functional Test for Improving Macular Degeneration

[0086] Healthy Kunming mice at 3 weeks old with a body weight of 19.4 - 20.1 g, half male and half female, were selected. Each mouse was fed 5 g of high-fat diet per day. After 5 months, a high-fat-induced macular degeneration model was successfully established. The mice were randomly divided into groups of 10 each. The AMX-EYE-03 group continued to be fed a normal diet for 1 month and was intragastrically administered 0.3 mL of an aqueous suspension of AMX-EYE-03 at a dose of 6 mg AMX-EYE-03 / 30 g body weight per day; the blank control group continued to be fed a normal diet for 1 month and was intragastrically administered 0.3 mL of normal saline per day. After the normal diet feeding, the mice were sacrificed, the eyeballs were removed, tissue sections were prepared and stained. Color images of the sections were continuously captured using a color digital camera. 10 images were randomly selected from each specimen for analysis, and the thickness of Bruch's membrane was measured. The average value of 100 measurements (10 times for each image) was taken for each specimen. The average value of each animal within the group was taken for each group. The results are shown in Table 2. Compared with other groups, the AMX-EYE-03 group could significantly reduce the thickness of Bruch's membrane, and there were statistically significant differences (P < 0.05). It was indicated that the composition AMX-EYE-03 could be used for macular degeneration.

[0087] Table 2. Macular Degeneration Test

[0088] Bruch membrane thickness / μm AMX-EYE-01 group 1.45 AMX-EYE-02 group 1.45 AMX-EYE-03 group 1.40 Blank control group 1.47

[0089] Example 4: Partial Active Ingredients of the Composition AMX-EYE-03

[0090] The solid powder composition AMX-EYE-03 prepared by the method described in Example 1 was taken, stirred at room temperature for 5 h in a mixed solvent of ethanol - isopropanol - water (volume ratio 3:1:3) with 20 times the weight, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was separated and purified using an InfinityLab Pursuit XRs C18 Preparative HPLC Column (gradient elution with acetonitrile - water) to obtain polypeptide AMX-EYE-P1 (SEQ ID NO:1) with a mass percentage of 0.02% in the composition AMX-EYE-03 and polypeptide AMX-EYE-P2 (SEQ ID NO:2) with a mass percentage of 0.03%. The structural characterization data of polypeptide AMX-EYE-P1 and AMX-EYE-P2 are the same as those in the following examples.

[0091] Polypeptide AMX-EYE-P1 (SEQ ID NO:1): SRTHANNVKGGDYAEVRLVDQETHNK. Polypeptide AMX-EYE-P2 (SEQ ID NO:2): NRQTHSGKSTDNQEGARVYHSTA.

[0092] Example 5: Preparation and Characterization of Polypeptides

[0093] The crude products of polypeptide AMX-EYE-P1 and AMX-EYE-P2 were obtained respectively by Fmoc solid-phase synthesis method using a polypeptide synthesizer (Aurora, VERSA 110).

[0094] The crude products were dried at 30 °C under vacuum conditions; separated and purified using an InfinityLab Pursuit XRs C18 Preparative HPLC Column (gradient elution with acetonitrile-water), and freeze-dried to obtain polypeptide AMX-EYE-P1 (SEQ ID NO:1) and AMX-EYE-P2 (SEQ ID NO:2), with HPLC purities of 96.3% and 95.8% respectively.

[0095] ESI-MS mass spectrometry analysis showed that its ion peaks corresponded to the molecular weights of the polypeptides of SEQ ID NO:1 and 2 sequences.

[0096] Polypeptide AMX-EYE-P1: 1 H NMR (400 MHz, DMSO-d6) δ8.92 - 7.98 (m, 36H), 7.67 (d, 8H), 7.43 (s, 6H), 7.22 (m, 2H), 7.01 - 6.92 (m, 2H), 6.85 (s, 2H), 6.63 - 6.54 (m, 2H), 5.02 (d, 2H), 4.89 - 3.78 (m, 39H), 3.54 - 1.48 (m, 74H), 1.27 - 0.89 (m, 23H).

[0097] Example 6: Functional Detection of Polypeptides

[0098] The same method as in Example 2 was used for testing, and the polypeptides were used instead of each composition for intragastric administration. The results are shown in Table 3. Compared with the other two groups, the AMX-EYE-P1 group could significantly reduce the iris congestion recovery time and conjunctival congestion recovery time, both with statistically significant differences (P < 0.001). It indicates that polypeptide AMX-EYE-P1 can significantly relieve eye fatigue.

[0099] Table 3. Functional Test for Relieving Visual Fatigue

[0100]

[0101]

[0102] Example VII: Preparation of Preparations

[0103] Preparation Example 1 of Preparations: The composition AMX-EYE-03 prepared by the method described in Example 1 was irradiated for sterilization, granulated, dried, and sized to obtain granules.

[0104] Preparation Example 2 of Preparations: The composition AMX-EYE-03 prepared by the method described in Example 1 was irradiated for sterilization, granulated, dried, and sized to obtain composition granules; the composition granules were filled into soft capsule shells, with 200 mg of the drug filled in each capsule shell to obtain soft capsules; the composition granules were filled into hard capsule shells, with 200 mg of the drug filled in each capsule shell to obtain hard capsules.

[0105] Preparation Example 3 of Preparations: The composition AMX-EYE-03 prepared by the method described in Example 1 was irradiated for sterilization and irradiated with 60Co, and the irradiation dose was 3 kGy; 4% by weight of ethanol with a concentration of 70% (V / V) was added to the sterilized mixture, and after being made into a soft material, it was granulated through a 12-mesh sieve; dried at 55 °C for 3 hours; sized to obtain granules.

[0106] Preparation Example 4 of Preparations: The composition AMX-EYE-03 prepared by the method described in Example 1 was irradiated for sterilization and irradiated with 60Co, and the irradiation dose was 3 kGy; 4% by weight of ethanol with a concentration of 70% (V / V) was added to the sterilized mixture, and after being made into a soft material, it was granulated through a 12-mesh sieve; dried at 55 °C for 3 hours; sized to obtain composition granules; the composition granules were filled into capsule shells, with 500 mg of the drug (composition granule mass) filled in each capsule shell to obtain capsules.

[0107] Preparation Example 5 of Preparations: The composition AMX-EYE-03 prepared by the method described in Example 1 was irradiated for sterilization and irradiated with 60Co, and the irradiation dose was 3 kGy; 4% by weight of ethanol with a concentration of 70% (V / V) was added to the sterilized mixture, and after being made into a soft material, it was granulated through a 12-mesh sieve; dried at 55 °C for 3 hours; sized to obtain composition granules; the composition granules were filled into capsule shells, with 600 mg of the drug (composition granule mass) filled in each capsule shell to obtain capsules.

[0108] Preparation Example 6 of the preparation: The composition AMX-EYE-03 prepared by the method described in Example 1 was sterilized by irradiation, irradiated with 60Co, and the irradiation dose was 3 kGy; 4% by weight of ethanol with a concentration of 70% (V / V) was added to the sterilized mixture, made into soft material and granulated through a 12-mesh sieve; dried at 55 °C for 3 hours; sized to obtain composition granules; the composition granules were filled into capsule shells, and each capsule shell contained 750 mg (the mass of the composition granules) of the medicine to obtain capsules.

[0109] Example 8. Acute toxicity test

[0110] Test sample: AMX-EYE-03 granules prepared by the method described in Preparation Example 3 of Example 7.

[0111] Experimental environmental conditions: Temperature 24.5 °C - 25.3 °C, humidity 54% - 60%.

[0112] Experimental animals: Healthy, adult clean-grade SD rats, with a body weight of 196 - 210 g, half male and half female, a total of 20 rats.

[0113] Experimental method: The maximum tolerated dose method was adopted. 100 g of the test sample was weighed and added with distilled water to 200 mL. The rats were gavaged 2 times at intervals of 4 hours within one day, and the dose for each time was 2 mL / 100 g body weight. Therefore, the total gavage dose on the same day was equivalent to 20000 mg / kg body weight. The rats were fasted for 20 hours before gavage and continuously observed for 7 days after gavage, and the poisoning manifestations and death conditions were recorded.

[0114] Test results: After gavage, the rats showed no obvious poisoning manifestations, and no rats died within 7 days. All the animals were sacrificed on the 8th day, dissected, and the surfaces of the main organs such as the liver, kidney, heart, lung, spleen, stomach, and intestine were smooth, and their sizes and shapes were normal, without any macroscopic pathological changes such as bleeding, congestion, and edema. The acute oral LD50 of male and female SD rats was greater than 20000 mg / kg body weight.

[0115] Conclusion: The acute toxicity test shows that AMX-EYE-03 is a non-toxic substance without toxic side effects.

[0116] Although the embodiments of the present disclosure have been described above, it can be understood that the above embodiments are exemplary and do not constitute a limitation to the present disclosure. Any modifications, equivalent replacements, and improvements made by those of ordinary skill in the art within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A polypeptide, characterized in that The polypeptide comprises the amino acid sequence shown in SEQ ID NO:

1.

2. The polypeptide according to claim 1, characterized in that The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

3. Use of the polypeptide according to claim 1 or 2 in the preparation of a drug for alleviating visual fatigue.

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