TW201932122A - Extraction method for cordycepin and use of cordycepin for manufacturing compositions for eye protection - Google Patents

Abstract

The present invention relates to an extraction method for cordycepin and a use of cordycepin for manufacturing compositions for eye protection by applying cordycepin to an individual in need. The extraction method comprises the steps of immersing a medium comprising Cordyceps militaries mycelia in an ethanol solution for 20 to 40 days to obtain a Cordyceps militaries crude extract; vacuuming, concentrating, and filtering the Cordyceps militaries crude extract to obtain a Cordyceps militaries extract; and conducting an absorption and separation process to the Cordyceps militaries extract by a resin to obtain cordycepin. The Cordycepin prepared in the present invention can be used for manufacturing artificial tears or eye drops.

Description

Method for extracting cordycepin and its use for preparing eye protection composition

The present invention relates to a method for extracting cordycepin and its use for preparing an eye protection composition.

In the eye disease, inflammation is a very common symptom, including pathogenic bacteria infection, eye conjunctiva or inflammation caused by corneal injury, etc., so the current common eye ophthalmic composition contains anti-inflammatory ingredients. In addition, dry eye syndrome is an eye disease caused by insufficient secretion of tears in the eyes or uneven distribution of tears, causing dryness of the surface of the eyeball; symptoms include dry eyes, itchy eyes, foreign body sensation, burning, sticky secretions, fear Light, phobia, and even symptoms that stimulate the secretion of reflexive tears; in addition, if the eyeballs are not kept moist, it will increase the chance of eye infection or injury.

The causes of dry eye syndrome are quite numerous, including air pollution, low ambient humidity, dry eye caused by drugs or diseases, and poor habits or eye habits. In addition, in addition to the lack of wetness of the eyes, dry eye patients are often accompanied by symptoms of inflammation, which leads to immune disorders. Therefore, if only the eye moisturizing component is provided without further treatment of the inflammatory condition, the symptoms of dry eye syndrome cannot be effectively treated or alleviated. The current drug for the treatment of dry eye syndrome is cyclosporine, the main pharmacological effect of which is immunosuppressive agent; but cyclosporin is a low water-soluble substance, so it can only be prepared into a cream or gel. Use, however, cream or gel can affect vision, is not suitable for daytime use, and cream or gel-based topical drugs are relatively inconvenient to use.

Nowadays, the inventors still have insufficient external composition for relieving or treating eye diseases, especially ocular inflammation-related diseases or dry eye syndrome, so they have many years of experience in designing, developing and practical production of related industries. The present invention has been developed based on this.

The main object of the present invention is to provide a method for extracting cordycepin and its use for preparing an eye protection composition; an effective dose of Cordycepin is administered to a desired individual to achieve eye protection.

The invention also provides a preparation method of a cordycepin-containing extract, comprising the following steps: Step 1: immersing a medium containing Cordyceps militaries mycelium in an ethanol solution for 20-40 days to obtain a Cordyceps militaris ethanol. Crude extract; Step 2: The crude ethanol extract of Cordyceps militaris is concentrated under vacuum and filtered, and the impurities are filtered to obtain the extract of Cordyceps militaris; and Step 3: the extract of Cordyceps militaris is subjected to an adsorption separation procedure with a resin to The cordycepin-containing extract of the present invention is obtained.

In one embodiment of the invention, the eye protection composition comprises artificial tears, eye drops or ophthalmic gels.

In one embodiment of the present invention, the mycelium-containing mycelium culture medium is immersed in a 75% ethanol solution for 30 days.

In one embodiment of the present invention, cordycepin inhibits microbial growth, reduces inflammatory response, modulates immune cell activity, and inhibits angiogenesis to achieve eye protection.

Accordingly, the present invention discloses the use of cordycepin for eye protection and discloses a method for extracting a large amount of cordycepin to provide a natural source, low toxicity and multi-effect eye protection composition.

For a more complete and clear disclosure of the technical content, the purpose of the invention and the effects thereof achieved by the present invention, the following detailed description is provided, and the reference drawings disclosed herein are also referred to:

First, the cultivation method of Cordyceps militaries

Cordyceps militaries used in this embodiment are Cordyceps militaris number BCRC 32219; the culture method is briefly described as follows: After inoculation of Cordyceps militaris in a culture solution, shake flask culture at 22 ° C to 28 ° C for 7 days to obtain The liquid culture of Cordyceps militaris, the culture temperature used in this experiment is 24 ° C; after a solid medium is autoclaved, the liquid strain of Cordyceps militaris is inoculated into the solid medium and placed in a culture vessel at 22 ° C. ~28 °C culture; first 7 days of dark cultivation, and then 35 days of bright cultivation, the culture temperature used in this experiment is 24 ° C; see the first figure, the cultivation of Cordyceps militaris can grow Cordyceps militaris fruit body on the medium, The medium also contains Cordyceps mycelium. The above solid medium is prepared by uniformly mixing 43.5 wt% of wheat, 0.013 wt% of soybean powder, 0.003 wt% of sucrose, and the remaining percentage of pure water, followed by autoclaving.

2. Preparation method of cordycepin-containing extract

The cultured Cordyceps militaris fruit body is separated from the medium containing the Cordyceps militaris mycelium, and frozen at -20 ° C for several days, taken out, pulverized into small agglomerates, and then separately immersed in a 75% ethanol solution, and Soak for 30 days at room temperature to obtain a crude ethanol extract of Cordyceps militaris; concentrate the crude ethanol extract of Cordyceps militaris in vacuum under reduced pressure, and then filter to remove impurities in the crude extract of Cordyceps militaris ethanol to obtain a Cordyceps militaris extract. Liquid; the Cordyceps militaris extract is further subjected to an adsorption separation step with a macroporous resin to prepare the cordycepin-containing extract of the present invention, wherein the macroporous resin may be, but not limited to, HPD722, HPD750, AB-8 or NKA-II Aperture resin.

The Cordyceps sinensis fruit body and the medium containing the Cordyceps militaris mycelium were subjected to the analysis of the cordycepin content of the cordycepin extract prepared in the above procedure, and the result showed that the cordycepin content contained in the Cordyceps militaris fruit body was 2.87 wt%, including The medium of Cordyceps militaris mycelium contained 1.2% by weight of cordycepin. However, the total weight of cordycepin produced by mycelial medium was higher than that of the fruiting body by 15 times of the total weight of chlorophyll.

Cordycepin, also known as 3'-Deoxyadenosine, is a water-soluble nucleoside antibiotic that inhibits a wide range of bacteria or viruses and has a broad spectrum of antibiotic properties. In addition, Cordycepin is also resistant. Inflammation, regulation of immune cell activity and inhibition of angiogenesis; in addition, cordycepin has very low cytotoxicity and is almost non-cytotoxic, and human adenosine deaminase (ADA) can decompose cordycepin, so it has no use. Cordycepin remains in the human body.

The main active ingredient used in the treatment of dry eye syndrome is cyclosporine, the main pharmacological effect of which is immunosuppressive agent; however, cyclosporin is a low water-soluble substance, so it can only be prepared into ointment. Or the use of a gel is quite inconvenient as an external medicine or health care product for the eye. Due to its excellent water solubility, it also has immunomodulatory activity, broad-acting antibacterial activity, etc., so it is very suitable for the preparation of aqueous preparations for eye-use products to improve the convenience of use; and Cordyceps of the present invention It is extracted from natural products and has low toxicity. It is safer to use.

In addition, because cordycepin has immunomodulatory effects, it can also be used as an immunosuppressant and an antibacterial agent after corneal transplantation to reduce postoperative rejection and infection.

The method for extracting cordycepin of the present invention and its use for preparing an eye-protecting composition can indeed achieve the intended use efficiency by the above-disclosed examples, and the present invention has not been disclosed before the application, and has been fully met. The provisions and requirements of the Patent Law.爰Issuing an application for a patent for invention in accordance with the law, and asking for a review, and granting a patent, is truly sensible.

However, the above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention; it is a person skilled in the art, and other aspects of the present invention. Modifications or modifications are considered to be within the scope of the design of the invention.

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First: Schematic diagram of artificial culture of Cordyceps militaris fruit bodies and culture medium.

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Claims (8)

A use of Cordycepin for the preparation of an eye protection composition is to administer an effective amount of cordycepin to a desired individual for the purpose of eye protection. The use of claim 1, wherein the eye protection composition comprises artificial tears or eye drops. The use according to claim 1, wherein the cordycepin inhibits microbial growth, reduces inflammatory response, modulates immune cell activity, and inhibits angiogenesis to achieve eye protection. A method for preparing Cordycepin extract comprises the following steps: Step 1: immersing a medium containing Cordyceps militaries mycelium in an ethanol solution for 20-40 days to obtain a Cordyceps militaris ethanol Crude extract; Step 2: The crude ethanol extract of Cordyceps militaris is concentrated under vacuum and filtered, and the impurities are filtered to obtain an extract of Cordyceps militaris; and Step 3: the extract of Cordyceps militaris is subjected to an adsorption separation process with a resin. The cordycepin-containing extract is obtained. The preparation method according to claim 4, wherein the frozen culture medium containing Cordyceps militaris is immersed in a 75% ethanol solution for 30 days. A use of a cordycepin extract obtained by the preparation method according to claim 4 or 5 for the preparation of an eye-protecting composition, wherein an effective amount of the cordycepin-containing extract is administered to a desired individual, To achieve the effect of eye protection. The use of claim 6, wherein the eye protection composition comprises artificial tears or eye drops. The use according to claim 6, wherein the cordycepin-containing extract inhibits microbial growth, reduces inflammatory response, modulates immune cell activity, and inhibits angiogenesis to achieve eye protection.