Seabuckthorn extract for prophylactic treatment of cataract and method of preparation thereof

Abstract

This invention relates to the field of cataract treatment technology, and discloses a sea buckthorn extract for the prevention and treatment of cataracts and its preparation method, comprising: step one, extraction; step two, concentration; step three, dissolution and loading; step four, washing with water to remove impurities; step five, elution with alcohol; and step six, concentration and drying. This sea buckthorn extract for the prevention and treatment of cataracts, through an optimized preparation process combining alcohol extraction and macroporous adsorption resin purification, directionally enriches the effective components, mainly flavonoids and polyphenols, from sea buckthorn medicinal materials. The resulting sea buckthorn extract has a clear group of active ingredients, overcoming the shortcomings of traditional crude sea buckthorn extracts, such as mixed components and difficulty in quality control. This provides a reliable technical guarantee for industrial production and drug quality control. Furthermore, since the sea buckthorn extract is derived from a natural medicinal and edible plant, compared with chemically synthesized drugs, it has less irritation to ocular tissues and extremely low toxicity, making it suitable for long-term use.

Description

Technical Field

[0001] This invention relates to the field of cataract treatment technology, and in particular to a sea buckthorn extract for the prevention and treatment of cataracts and its preparation method. Background Technology

[0002] Cataracts are the leading cause of blindness worldwide. The pathological essence of cataracts lies in the denaturation and aggregation of lens proteins caused by various factors (such as aging, genetics, metabolic abnormalities, trauma, radiation, etc.), which leads to clouding and prevents light from passing normally to the retina, resulting in progressive vision loss or even complete blindness. With the continued acceleration of the global population aging process, the prevalence of cataracts is increasing year by year, bringing a heavy economic burden and social care pressure to public health systems in various countries.

[0003] Currently, the only definitive and clinically recognized method to effectively restore vision in cataract patients is surgical treatment, namely phacoemulsification lens extraction combined with intraocular lens implantation. Although this procedure has become increasingly mature, it still faces many practical bottlenecks in clinical application: First, the uneven distribution of medical resources means that many patients in developing countries, remote areas, and economically underdeveloped regions suffer delays in treatment due to limited financial resources or inability to access timely surgical care. Second, as an invasive procedure, surgery itself carries certain risks of postoperative complications, such as posterior cataract (posterior capsulitis), endophthalmitis, and macular edema, which directly affect the patient's long-term visual quality and quality of life.

[0004] Therefore, developing drug therapies that can effectively delay, prevent, or even reverse the progression of lens opacity, especially for early intervention in cataract development or for drugs to inhibit the formation of posterior capsule opacification after surgery, has always been a research hotspot and a major challenge in the field of ophthalmology. Currently, commercially available cataract treatment drugs mainly include some chemically synthesized preparations and some traditional Chinese medicine compound preparations; However, existing drugs generally have the following drawbacks: 1) The efficacy is uncertain. Most drugs can only play a limited role in delaying the disease and are difficult to fundamentally reverse the pathological changes of lens opacity. 2) It has strong toxic side effects and local irritation. Long-term use of ocular medication can easily cause adverse reactions such as ocular surface damage and allergies. 3) Low bioavailability, making it difficult for the drug to penetrate the corneal barrier and reach an effective therapeutic concentration in the aqueous humor and lens; 4) The effective components of natural medicines / traditional Chinese medicines are unclear, and the material basis is unclear, which makes quality control difficult, and the efficacy varies greatly between batches, making it difficult to meet the requirements of modern drug standardization and precision. Summary of the Invention

[0005] Given the uncertain efficacy of existing drugs, most of which only provide limited delay and cannot fundamentally reverse the pathological changes of lens opacity; their strong toxic side effects and local irritation, with long-term ocular use easily causing ocular surface damage, allergies, and other adverse reactions; their low bioavailability, making it difficult for drugs to penetrate the corneal barrier and reach effective therapeutic concentrations in the aqueous humor and lens; and the unclear effective components of natural drugs / traditional Chinese medicines, leading to difficulties in quality control and large batch-to-batch differences in efficacy, making it difficult to meet the requirements of modern drug standardization and precision, this invention was proposed.

[0006] Therefore, the purpose of this invention is to provide a sea buckthorn extract for the prevention and treatment of cataracts. The purpose is that sea buckthorn, a plant of the Elaeagnaceae family, is a medicinal and edible plant rich in flavonoids, polyphenols, vitamins, amino acids, and trace elements, possessing strong antioxidant, anti-inflammatory, and anti-aging pharmacological activities. Traditional medicine and modern nutrition have records of sea buckthorn's use in eye health care, but existing technologies mostly focus on the edible health benefits of crude extracts or juices from sea buckthorn fruits. For the specific indication of cataracts, there is a lack of a sea buckthorn extract and its pharmaceutical preparation with a clear material basis, a stable and controllable preparation process, and systematic pharmacodynamic verification. This would allow for the targeted enrichment of effective parts or components directly related to cataract prevention and treatment from sea buckthorn, and then effective intraocular delivery through a scientifically formulated preparation.

[0007] To solve the above-mentioned technical problems, the present invention provides the following technical solution: a sea buckthorn extract for the prevention and treatment of cataracts, comprising the following steps: Step 1, Extraction: Take sea buckthorn medicinal material, crush or not crush it, add 6 to 12 times the weight of the medicinal material in 50% to 90% ethanol, and extract by heating and reflux or ultrasonic extraction 2 to 5 times, and combine the extracts; Step 2, Concentration: Concentrate the combined extract obtained in Step 1, and recover the ethanol until there is no ethanol odor, to obtain sea buckthorn extract; Step 3, Dissolving and Loading: Dissolve the sea buckthorn extract obtained in Step 2 in water and load it onto a macroporous adsorption resin column. Step 4: Washing to remove impurities: Elute the macroporous adsorption resin column with purified water until the eluent is colorless; Step 5: Eluent elution: Elute the macroporous adsorption resin column with 50% to 90% ethanol until the eluent is colorless, and collect the ethanol eluent. Step 6, Concentration and Drying: The ethanol eluent collected in Step 5 is concentrated and dried to obtain the sea buckthorn extract used for the prevention and treatment of cataracts.

[0008] In a preferred embodiment of the present invention, the macroporous adsorption resin in step three is selected from one of the following macroporous adsorption resins: D101, AB-8, DM130, and X-5.

[0009] In a preferred embodiment of the present invention, the sea buckthorn extract in step three is in a weight ratio of 1:10 to 1:50 based on the weight of the dry extract to the wet macroporous adsorption resin.

[0010] As a preferred embodiment of the present invention, the sea buckthorn medicinal material in step one is one or more of sea buckthorn fruit, sea buckthorn leaves or sea buckthorn seeds; The concentration described in step two is atmospheric pressure concentration or vacuum concentration.

[0011] In a preferred embodiment of the present invention, the sea buckthorn extract is prepared by the method described above for preparing sea buckthorn extract for the prevention and treatment of cataracts.

[0012] As a preferred embodiment of the present invention, the use of sea buckthorn extract for the prevention and treatment of cataracts in the preparation of a medicament for the prevention and / or treatment of cataracts is described.

[0013] As a preferred embodiment of the present invention, the cataract includes age-related cataract, diabetic cataract, complicated cataract, traumatic cataract, or posterior cataract.

[0014] In a preferred embodiment of the present invention, the drug is an intraocular preparation.

[0015] As a preferred embodiment of the present invention, the intraocular preparation is an eye drop, an ophthalmic gel, an eye ointment, or an intraocular injection.

[0016] As a preferred embodiment of the present invention, a pharmaceutical composition for the prevention and / or treatment of cataracts is characterized by comprising sea buckthorn extract and pharmaceutically acceptable excipients.

[0017] Compared with the prior art, the present invention has at least the following beneficial effects: 1. This invention optimizes the preparation process by combining alcohol extraction with macroporous adsorption resin purification, thereby directionally enriching the effective components, mainly flavonoids and polyphenols, from sea buckthorn. The resulting sea buckthorn extract has a clear group of active ingredients, overcoming the shortcomings of traditional crude sea buckthorn extracts, such as mixed components and difficulty in quality control. This provides a reliable technical guarantee for industrial production and drug quality control. Moreover, since sea buckthorn extract is derived from a natural medicinal and edible plant, it has less irritation to eye tissues and extremely low toxicity compared to chemically synthesized drugs, making it suitable for long-term use. It exerts its protective effect through multi-target antioxidant and free radical scavenging mechanisms, and is especially suitable for the intervention and treatment of early cataracts and the prevention of postoperative secondary cataracts.

[0018] 2. This invention uses ethanol reflux extraction or ultrasonic extraction technology, combined with a specific type of macroporous adsorption resin for purification. By precisely limiting key process parameters such as the material-liquid ratio, ethanol concentration, number of extractions, and resin dosage, it achieves efficient extraction and enrichment of the target active ingredients. Moreover, the process route is simple, easy to operate, and has good repeatability, making it easy to promote and apply in the pharmaceutical industry.

[0019] 3. This invention establishes in vitro high glucose and aging models of lens epithelial cells, as well as in vivo diabetic cataract animal models and age-related cataract animal models, to systematically verify that the sea buckthorn extract of this invention can significantly improve the survival rate of lens epithelial cells and reduce intracellular oxidative stress levels (such as reducing MDA content and increasing GSH-PX activity), thereby effectively delaying or inhibiting the occurrence and development of lens opacity.

[0020] 4. The sea buckthorn extract of the present invention can be formulated into various intraocular preparations such as eye drops, ophthalmic gels, ophthalmic ointments, and intraocular injections according to clinical needs. It can flexibly adapt to the medication habits and treatment plans of different patient groups and has good clinical applicability and market application prospects. Attached Figure Description

[0021] Figure 1 This is a flowchart illustrating the steps of preparing the sea buckthorn extract for the prevention and treatment of cataracts according to the present invention. Figure 2 This is a dosage comparison chart of the sea buckthorn extract used in the prevention and treatment of cataracts according to the present invention. Detailed Implementation

[0022] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0023] This invention provides a sea buckthorn extract for the prevention and treatment of cataracts and its preparation method. The sea buckthorn extract is derived from the Elaeagnaceae plant Hippophae rhamnoides. A specific alcohol extraction process combined with macroporous adsorption resin purification is used to directionally enrich the effective components, mainly flavonoids and polyphenols, from the sea buckthorn medicinal material. The obtained sea buckthorn extract exhibits clear antioxidant activity, effectively protecting lens epithelial cells and delaying the progression of lens opacity. It can be used to prepare drugs for the prevention and / or treatment of cataracts.

[0024] General preparation process The general process for preparing sea buckthorn extract according to the present invention is as follows: Step 1: Extraction Take sea buckthorn medicinal material, crush it or not, add 6 to 12 times the weight of the medicinal material in 50% to 90% ethanol, and extract by heating and reflux or ultrasonic extraction 2 to 5 times, and combine the extracts.

[0025] The sea buckthorn medicinal material can be selected from one or more of the following: sea buckthorn fruit, sea buckthorn leaves, or sea buckthorn seeds. The extraction method can be flexibly selected according to production conditions: heating reflux extraction equipment is simple and easy to operate, suitable for large-scale production; ultrasonic extraction has high efficiency and low extraction temperature, which is beneficial for the retention of heat-sensitive components.

[0026] Step Two: Concentration The combined extract obtained in step one was concentrated, and the ethanol was recovered until there was no ethanol odor, to obtain sea buckthorn extract.

[0027] Concentration can be achieved using either atmospheric pressure concentration or vacuum concentration. Vacuum concentration, which removes the solvent rapidly at lower temperatures, is the preferred method as it helps retain the heat-sensitive active ingredients in the extract.

[0028] Step 3: Dissolve and load the sample After dissolving the sea buckthorn extract obtained in step two in water, the sample was loaded onto a macroporous adsorption resin column.

[0029] The macroporous adsorption resin is selected from one of the following types: D101, AB-8, DM130, and X-5. These types of macroporous adsorption resins exhibit good adsorption selectivity and desorption performance for flavonoids and polyphenols. The weight ratio of sea buckthorn extract (dry extract) to wet macroporous adsorption resin is 1:10 to 1:50.

[0030] Step 4: Washing to remove impurities The macroporous adsorption resin column was eluted with purified water until the eluent was colorless.

[0031] The main purpose of this step is to remove water-soluble impurities, such as sugars, organic acids, and inorganic salts, from the sea buckthorn extract that have not been adsorbed by the resin, in order to enrich the target active ingredients and improve the purity of the extract.

[0032] Step 5: Alcohol Elution The macroporous adsorption resin column was eluted with 50%–90% ethanol until the eluent was colorless, and the ethanol eluent was collected.

[0033] This step utilizes ethanol's excellent elution ability for flavonoids and polyphenols adsorbed on the resin to desorb the target active components from the resin. The concentration of ethanol used is kept close to that used in the extraction step one to ensure elution efficiency.

[0034] Step Six: Concentration and Drying The ethanol eluent collected in step five is concentrated and dried to obtain sea buckthorn extract for the prevention and treatment of cataracts.

[0035] The drying method can be conventional methods such as vacuum drying, spray drying or freeze drying to obtain solid powdered sea buckthorn extract that is easy to process into subsequent formulations.

[0036] Example 1 (Preparation of Seabuckthorn Extract A) (1) Take sea buckthorn medicinal material and crush it into coarse powder. Add 8 times the amount of 70% ethanol to the medicinal material and extract it three times with ultrasound. Combine the extracts and concentrate them under reduced pressure to recover the ethanol until there is no ethanol taste, and obtain sea buckthorn extract.

[0037] (2) Dissolve the sea buckthorn extract and wet macroporous adsorption resin in water at a ratio of 1:20 (dry extract weight to wet resin weight). Then, load the mixture onto a D101 macroporous adsorption resin column and elute with purified water until the eluent is colorless. Then, elute with 70% ethanol until the eluent is colorless. Collect the ethanol eluent, concentrate and dry the eluent to obtain the sea buckthorn extract, which is named sea buckthorn extract A.

[0038] Example 2 (Preparation of Seabuckthorn Extract B) (1) Take sea buckthorn medicinal material and crush it into coarse powder. Add 12 times the amount of 90% ethanol to the medicinal material and extract it twice by heating and reflux. Combine the extracts and concentrate them under reduced pressure to recover the ethanol until there is no ethanol taste, and obtain sea buckthorn extract.

[0039] (2) Dissolve the sea buckthorn extract and wet macroporous adsorption resin in water at a ratio of 1:50 (dry extract weight to wet resin weight). Then, load the sample onto an X-5 type macroporous adsorption resin column and elute with purified water until the eluent is colorless. Then, elute with 90% ethanol until the eluent is colorless. Collect the ethanol eluent, concentrate and dry the eluent to obtain the sea buckthorn extract, which is named sea buckthorn extract B. Example

[0040] (1) Take sea buckthorn medicinal material and crush it into coarse powder. Add 6 times the amount of 50% ethanol to the medicinal material and extract it 5 times with ultrasound. Combine the extracts and concentrate them under reduced pressure to recover the ethanol until there is no ethanol taste, and obtain sea buckthorn extract.

[0041] (2) Dissolve the sea buckthorn extract and wet macroporous adsorption resin in water at a ratio of 1:10 (dry extract weight to wet resin weight). Then, load the sample onto an AB-8 macroporous adsorption resin column and elute with purified water until the eluent is colorless. Then, elute with 50% ethanol until the eluent is colorless. Collect the ethanol eluent, concentrate and dry the eluent to obtain the sea buckthorn extract. Example

[0042] (1) Take sea buckthorn medicinal material and crush it into coarse powder. Add 10 times the amount of 70% ethanol to the medicinal material and extract it twice with ultrasound. Combine the extracts and concentrate them under reduced pressure to recover the ethanol until there is no ethanol taste, and obtain sea buckthorn extract.

[0043] (2) Dissolve the sea buckthorn extract and wet macroporous adsorption resin in water at a ratio of 1:30 (dry extract weight to wet resin weight). Then, load the mixture onto a DM130 macroporous adsorption resin column and elute with purified water until the eluent is colorless. Then, elute with 70% ethanol until the eluent is colorless. Collect the ethanol eluent, concentrate and dry the eluent to obtain the sea buckthorn extract.

[0044] Pharmacological experimental methods and results This invention verifies the cataract prevention and treatment efficacy of sea buckthorn extract through the following pharmacological experimental model (reference). Figure 2 The experimental group settings are as follows: Normal control group: Healthy animals or normally cultured cells were given an equal amount of solvent as a control; Diabetic model group: A diabetic cataract animal model was established by intraperitoneal injection of streptozotocin (STZ) combined with high-fat diet, or a lens epithelial cell oxidative damage model was induced by high-glucose culture medium. Low-dose group of sea buckthorn extract A: Low-dose sea buckthorn extract A was administered in addition to the model. High-dose group of sea buckthorn extract A: High doses of sea buckthorn extract A were administered in addition to the model. Low-dose group of sea buckthorn extract B: Low-dose sea buckthorn extract B was administered in addition to the model. High-dose group of sea buckthorn extract B: High doses of sea buckthorn extract B were administered based on the model.

[0045] (a) In vitro experiments In vitro high glucose models and hydrogen peroxide-induced aging models of lens epithelial cells (SRA01 / 04, HLE-B3) were established. Following the experimental group settings described above, cells were treated with different concentrations of the present invention's sea buckthorn extract A or sea buckthorn extract B. Cell viability was detected using the MTT assay, and intracellular oxidative stress-related indicators (MDA content, GSH-PX activity, SOD activity) were detected using biochemical reagent kits.

[0046] Experimental results showed that, compared with the diabetic model group, both low-dose and high-dose groups of sea buckthorn extract A and sea buckthorn extract B dose-dependently increased the survival rate of lens epithelial cells under high glucose and oxidative stress conditions, significantly reduced intracellular MDA levels, and increased GSH-PX and SOD activities. The high-dose group showed better results than the low-dose group. This indicates that the sea buckthorn extract of the present invention can effectively protect lens epithelial cells through an antioxidant pathway, delaying or preventing the occurrence and development of cataracts.

[0047] (II) In vivo experiments Diabetic cataract model: A rat model of diabetic cataract was established by intraperitoneal injection of streptozotocin (STZ) combined with a high-fat diet. The rats were divided into groups as described above: the normal control group received an equal volume of physiological saline; the diabetic model group received an equal volume of solvent; and each treatment group received eye drops prepared from sea buckthorn extract A or sea buckthorn extract B at low or high doses, respectively, for 8 consecutive weeks. The degree of lens opacity was observed and graded using a slit-lamp microscope, and the MDA content and antioxidant enzyme activity within the lens were also measured.

[0048] The experimental results showed that, compared with the diabetic model group, all doses of sea buckthorn extract A and sea buckthorn extract B could significantly delay the progression of lens opacity in diabetic rats, reduce the MDA content in the lens, and increase the activity of antioxidant enzymes, with the high-dose group showing more significant improvement.

[0049] Age-related cataract model: An age-related cataract model was established in rats using ultraviolet irradiation. Following the experimental groupings described above, each treatment group received eye drops prepared from either sea buckthorn extract A or sea buckthorn extract B, administered continuously for two weeks after eye opening. The incidence and degree of lens opacity were observed.

[0050] Experimental results showed that, compared with the model group, all dose groups of sea buckthorn extract A and sea buckthorn extract B could significantly reduce the incidence of sodium selenite-induced cataracts and alleviate the degree of lens opacity, exhibiting a good dose-response relationship.

[0051] Formulation preparation example The sea buckthorn extract of this invention can be formulated into various intraocular preparations with pharmaceutically acceptable excipients according to clinical needs. The following are typical examples of such preparations: Example of eye drop preparation: Take an appropriate amount of the sea buckthorn extract of this invention, dissolve it in water for injection, add an appropriate amount of sodium chloride to adjust the osmotic pressure, add an appropriate amount of ethylparaben as a preservative, add an appropriate amount of sodium hyaluronate as a thickener, adjust the pH value to 6.5-7.5, filter to remove bacteria, and aseptically dispense to obtain sea buckthorn extract eye drops.

[0052] Scope of application The sea buckthorn extract of this invention can be used to prevent and / or treat various types of cataracts, including but not limited to age-related cataracts, diabetic cataracts, complicated cataracts, traumatic cataracts, and secondary cataracts. Its mechanism of action is mainly achieved through scavenging free radicals, inhibiting oxidative damage to lens epithelial cells, and maintaining the stability of lens proteins.

[0053] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A method for preparing sea buckthorn extract for the prevention and treatment of cataracts, characterized in that, Includes the following steps: Step 1, Extraction: Take sea buckthorn medicinal material, crush or not crush it, add 6 to 12 times the weight of the medicinal material in 50% to 90% ethanol, and extract by heating and reflux or ultrasonic extraction 2 to 5 times, and combine the extracts; Step 2, Concentration: Concentrate the combined extract obtained in Step 1, and recover the ethanol until there is no ethanol odor, to obtain sea buckthorn extract; Step 3, Dissolving and Loading: Dissolve the sea buckthorn extract obtained in Step 2 in water and load it onto a macroporous adsorption resin column. Step 4: Washing to remove impurities: Elute the macroporous adsorption resin column with purified water until the eluent is colorless; Step 5, Alcohol Elution: Elute the macroporous adsorption resin column with 50% to 90% ethanol until the eluent is colorless, and collect the ethanol eluent. Step 6, Concentration and Drying: The ethanol eluent collected in Step 5 is concentrated and dried to obtain sea buckthorn extract for the prevention and treatment of cataracts.

2. The method for preparing sea buckthorn extract for the prevention and treatment of cataracts according to claim 1, characterized in that: The macroporous adsorption resin mentioned in step three is selected from one of the following macroporous adsorption resins: D101, AB-8, DM130, and X-5.

3. The method for preparing sea buckthorn extract for the prevention and treatment of cataracts according to claim 1, characterized in that: The sea buckthorn extract mentioned in step three has a weight ratio of 1:10 to 1:50 between the dry extract and the wet macroporous adsorption resin.

4. The method for preparing sea buckthorn extract for the prevention and treatment of cataracts according to claim 3, characterized in that: The sea buckthorn medicinal material mentioned in step one is one or more of the following: sea buckthorn fruit, sea buckthorn leaves, or sea buckthorn seeds; The concentration described in step two is atmospheric pressure concentration or vacuum concentration.

5. A sea buckthorn extract for the prevention and treatment of cataracts, characterized in that, The sea buckthorn extract is prepared by the method described in any one of claims 1 to 4.

6. The use of the sea buckthorn extract according to claim 5 for the prevention and treatment of cataracts in the preparation of a medicament for the prevention and / or treatment of cataracts.

7. The application of sea buckthorn extract for the prevention and treatment of cataracts according to claim 6, characterized in that: The cataracts include age-related cataracts, diabetic cataracts, complicated cataracts, traumatic cataracts, or secondary cataracts.

8. The application of sea buckthorn extract for the prevention and treatment of cataracts according to claim 7, characterized in that: The drug is an intraocular preparation.

9. The application of sea buckthorn extract for the prevention and treatment of cataracts according to claim 8, characterized in that: The intraocular preparation is an eye drop, ophthalmic gel, eye ointment, or intraocular injection.

10. A pharmaceutical composition for the prevention and / or treatment of cataracts, characterized in that, It comprises the sea buckthorn extract as described in claim 5 and pharmaceutically acceptable excipients.

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