CN113398252A - Eye drop rich in animal and plant active ingredients and preparation method thereof - Google Patents
Eye drop rich in animal and plant active ingredients and preparation method thereof Download PDFInfo
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- CN113398252A CN113398252A CN202110726104.0A CN202110726104A CN113398252A CN 113398252 A CN113398252 A CN 113398252A CN 202110726104 A CN202110726104 A CN 202110726104A CN 113398252 A CN113398252 A CN 113398252A
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/898—Orchidaceae (Orchid family)
- A61K36/8984—Dendrobium
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/28—Insulins
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
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- A61P27/02—Ophthalmic agents
- A61P27/12—Ophthalmic agents for cataracts
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/30—Extraction; Separation; Purification by precipitation
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
Abstract
The invention provides an eye drop rich in animal and plant active ingredients, which comprises 0.1-5 parts of active small molecular polypeptide, 5-15 parts of traditional Chinese medicine extract, 0.1-5 parts of lubricating and moisturizing enhancer, 0.1-0.5 part of preservative, 1-5 parts of osmotic pressure regulator, and the balance of normal saline, wherein the pH value is 6.0-8.0. The preparation process of the invention adopts various modern biological high-tech integration technologies, the active ingredients and active substances in the traditional Chinese medicine and animal tissues are retained to the greatest extent, the active substances have synergistic effect and can supplement the nutrition of eye nerve cells, and the extracted traditional Chinese medicine extract and the extracted active small molecular polypeptide have the molecular weight of less than 1000Da, can permeate various eye tissue barriers to reach the eyeground part, and are suitable for various eyeground part diseases such as eye nebula, eye dryness and tiredness, asthenopia, blurred vision, night blindness and the like.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to an eye drop rich in animal and plant active ingredients and a preparation method thereof.
Background
In daily office and life, with the great popularization and wide use of various electronic audio-visual equipment such as mobile phones, computers, televisions and the like, many people are easy to have excessive eye use and incorrect eye use problems in the long-term use process, and in addition, various harmful light such as ultraviolet rays, blue light and the like invade eyes, eye diseases such as blurred vision, visual cell damage, optic atrophy, macular degeneration of retina and the like are easy to cause. Because the harm caused by the diseases is large, timely prevention and treatment are required to be found out.
There are many nerve cells around the eye, which are very vulnerable. Optic atrophy is the damage of optic nerve lesions and myelin sheaths or retinal ganglion cells and axons thereof, which usually occurs from retina to outer side, resulting in the final outcome of nerve fiber loss and gliosis, and patients often have visual function deterioration and visual field loss or shrinkage due to degeneration and atrophy of nerve fibers.
Optic nerve disease refers to a disease of retinal ganglion cell axonal degeneration caused by various causes, and is mainly characterized by visual function impairment and papillary pallor. Optic atrophy is the end result of optic neuropathy. It is manifested by degeneration and disappearance of optic nerve fibers, conduction dysfunction, visual field change, and visual deterioration and loss. Clinically, the atrophy is divided into primary atrophy and secondary atrophy. The causes of the disease are that, in general, optic atrophy of children is caused by brain tumor or intracranial inflammation; young patients are mainly inherited; middle-aged patients are mostly optic neuritis or nerve trauma; the elderly are often associated with glaucoma or vascular causes.
Clinically, about 2/3 blinding eye diseases are associated with neovascular or inflammatory responses, such as macular degeneration, uveitis, etc. At present, aiming at the diseases clinically, the medicine administration modes such as systemic medicine administration, intravitreal injection or surgical implantation are mainly adopted, so that the effective components in the medicine can act on the fundus part, great pain is caused to patients clinically, and high cost and high treatment risk are required. However, due to the blood-ocular barrier, macromolecular drugs are difficult to penetrate into the tissues of the eye to achieve sufficient drug concentration in the eye, and exogenous macromolecular drugs tend to cause immunological damage to the ocular tissues.
Therefore, in the research of searching a noninvasive or minimally invasive administration method, the small molecular peptide drug which has biological activity, strong tissue penetrability, small toxic and side effect and homology of medicine and food is found, and the small molecular peptide drug has important significance for diagnosis and treatment of eye diseases. The bioactive peptide drug has small molecules and simple structure; the immunogenicity is small, and the possibility of inducing host immune response is low; has more advantages in the aspect of target penetration of eye tissues. These characteristics make the bioactive peptide drug have great potential advantages in clinical application of ophthalmology.
Disclosure of Invention
The invention aims to provide an eye drop rich in animal and plant active ingredients and a preparation method of the eye drop.
The eye drop is rich in various animal and plant active ingredients, can supplement nutrition of nerve cells of eyes, nourish the visual nervous system and optic nerve cells, enhance the activity of axon papilla nerve cells in iris, choroid and macular areas of retina, and inhibit or slow down macular degeneration of retina and optic atrophy; has the effects of relieving itching of eyes, relaxing ciliary muscle, relieving eye fatigue, eliminating dryness of eyes, and improving eyesight; is suitable for eye ground diseases such as nebula, dry eye, eye fatigue, blurred vision, and nyctalopia. The eye drop is safe, convenient and efficient, has no toxic or side effect, is easy for large-scale production, and can meet the needs of people.
An eye drop rich in animal and plant active ingredients comprises the following components in parts by weight: active small molecular polypeptide 0.1-5 parts, Chinese medicinal extract 5-15 parts, lubricant and moisture enhancer 0.1-5 parts, antiseptic 0.1-0.5 part, osmotic pressure regulator 1-5 parts, and physiological saline for balance, and pH is 6.0-8.0.
Furthermore, the eye drop rich in animal and plant active ingredients can also be added with insulin aiming at hyperglycemia patients, and the addition amount of the insulin in a finished product is 0.3-0.5 mg/mL.
Further, the lubricating and moisturizing enhancer comprises one or two or more of sodium hyaluronate, polyvinyl alcohol, tween 80, hydroxyethyl cellulose, sodium carboxymethyl cellulose, carbomer, polyoxyethylene 9-lauryl ether, polyoxyethylene 20-stearyl ether and glycerol.
Further, the preservative comprises one or two or more of borneol, menthol, polylysine, chitosan, chlorobutanol and borax.
Further, the osmotic pressure regulator includes, but is not limited to, one or two or more of sodium chloride, boric acid, potassium nitrate, glucose, and the like.
The active polypeptide is obtained by the following extraction process:
(1) taking healthy fresh animal tissues, tissue fluid or body fluid rich in active polypeptide, removing impurities, cleaning, cutting, and mashing with a tissue mashing machine;
(2) adding sodium chloride solution with equal mass volume, mixing uniformly, and grinding by a colloid mill to form homogenate;
(3) adding 1-3 times of sodium chloride solution, mixing, adding protease (including but not limited to pancreatin, papain, and complex enzyme) 1-2% of the weight of the raw materials, heating in water bath to 50-55 deg.C, adjusting pH to 7.5-8.5 with saturated calcium hydroxide, and hydrolyzing for 10-15 hr under uniform stirring;
(4) standing at-10-10 deg.C for 8-12h, wherein the precipitation method includes but is not limited to cold precipitation, isoelectric point precipitation, organic solvent precipitation or salting-out precipitation;
(5) taking out, centrifuging at 4000 and 10000r/min at 0-4 ℃ for 5-30min to remove protein precipitate, and collecting supernatant:
(6) filtering the supernatant through a filter material with the aperture of 0.22-1.2um to remove impurities and micro particles, and collecting a clarified liquid:
(7) ultrafiltering, nanofiltering, and concentrating at multiple gradient to obtain active small molecular polypeptide with molecular weight of below 1000Da, and freezing for storage;
the traditional Chinese medicine extract is obtained by the following extraction process:
(1) weighing 40-80g of dendrobium, 160g of cassia seed, 50-200g of medlar, 160g of wild chrysanthemum flower and 10-25g of pearl in turn according to the requirements in the formula; slicing, cutting, pulverizing, placing in an ultrasonic reactor, stirring and mixing;
(2) adding 1.5-3 times of pure water or deionized water, soaking for 0.5-2 hr, and mixing to obtain mixed medicinal liquid;
(3) treating the mixed liquid medicine obtained in the step (2) for 5-30min by ultrasonic wave with the frequency of 10-200KHz and the temperature kept at 50-60 ℃, filtering by using a 100-mesh filter screen or gauze, collecting precipitates to be treated, and collecting filtrate for later use;
(4) putting the precipitate in the step (3) into an ultrasonic reactor again, adding 1.5-3 times of pure water or deionized water, performing ultrasonic treatment for 5-30min at the ultrasonic frequency of 10-200kHz and the temperature of 50-60 ℃, filtering by using a 100-mesh filter screen or gauze, and collecting the filtrate for later use;
(5) mixing the filtrates collected in steps (3) and (4) to obtain Chinese medicinal extract, and refrigerating for use.
A method for preparing eye drop rich in animal and plant active ingredients comprises the following steps:
(1) preparation of a NaCl solution: weighing 0.5-1.5g of NaCl, adding deionized water to a constant volume of 100mL to obtain a NaCl solution;
(2) adding 0.1-5 parts of active polypeptide and 5-15 parts of Chinese medicinal extract into 100mL NaCl solution, dissolving, adjusting pH to 6.0-8.5, standing at 0-8 deg.C for 4-8 hr to obtain mixed solution of active polypeptide and Chinese medicinal extract;
(3) weighing 0.1-5 parts of lubricating and moisturizing reinforcing agent; sequentially adding the mixture into 500mL of pure water or deionized water, and stirring the mixture to dissolve the mixture to obtain a lubricating and moisturizing reinforcing agent solution;
(4) heating 100mL of purified water or deionized water to 40-50 ℃, adding 1-5 parts of osmotic pressure regulator, stirring for dissolving, and standing at room temperature to obtain the osmotic pressure regulator;
(5) weighing 1-10 parts of antiseptic materials such as borneol and the like, grinding into powder, and adding the powder into 100mL of NaCl solution for uniformly mixing to obtain an antiseptic;
(6) mixing the active polypeptide and Chinese medicinal extract mixed solution prepared in steps (2), (3), (4) and (5), lubricant and moisture-keeping enhancer, antiseptic and osmotic pressure regulator, adding physiological saline to 1000mL, adjusting pH to 6.0-8.0, standing at 0-4 deg.C for 24-48h, and making into basic eye medicine;
(7) filtering and sterilizing by using filter materials with the pore diameters of 1.0um, 0.45um and 0.15um in a grading manner under an aseptic clean environment; sterile subpackaging to obtain the eye drop rich in animal and plant active ingredients.
Furthermore, the eye drop rich in animal and plant active ingredients prepared by the invention can be prepared into other dosage forms such as different injections, aqueous solutions, atomizing agents, powders, gels, ointments, liquid dressings, eye patches, freeze-dried powders and other administration modes such as administration devices or apparatuses by freeze-drying, spray-drying or adding excipients and the like.
The dendrobium in the formula has the effects of nourishing yin and improving eyesight, is a special health-care product for protecting eyes, and can prevent the vision from being degraded and reduce various ophthalmic diseases such as cataract glaucoma, myopia and the like when people eat the dendrobium for a long time.
The cassia seed in the formula is a commonly used medicine for clearing liver and improving vision, is also called as 'pupil reduction' because of obvious vision improving effect, is mainly used for clinically treating symptoms such as red and swollen eyes, pain, blurred vision, headache and dizziness, intestinal dryness and constipation and the like caused by liver meridian heat and liver fire flaming, and has good effect.
The medlar in the formula has the effects of nourishing kidney yin, clearing liver and improving vision. As long as it is said from Shen nong Ben Cao Jing, the medlar has the effects of being taken for a long time, being light and not old, resisting cold and summer heat, and delaying senility, so it has another name named 'Lao Zi'. The medlar contains various amino acids, and special nutrient components such as betaine, baccatin and the like. The property of Chinese wolfberry is also sweet and neutral, and then enters liver and kidney meridians, and the efficacy is mainly reflected in nourishing liver and kidney, replenishing vital essence and improving eyesight, clearing liver heat, and being helpful to improve blurred vision and maintain eyesight.
The wild chrysanthemum flower in the formula has the effects of clearing away heat and toxic materials, dispelling wind and heat, calming the liver and improving eyesight, and is mainly used for treating carbuncle, cellulitis and furuncle, sore throat, conjunctival congestion and swelling, headache and vertigo and the like. For fever, headache, dim vision due to liver and kidney deficiency, and headache and vertigo due to liver fire flaming up, it can be used with Jue Ming Zi to protect eyes.
The pearl in the formula of the invention has sweet and salty taste and cold property, and enters heart and liver meridians. Margarita has effects of clearing liver heat, improving eyesight, removing nebula, tranquilizing, arresting convulsion, clearing away heat and toxic materials, and can be used for treating eye diseases such as vexation, insomnia, dreaminess, amnesia, restlessness, pyocutaneous disease, ulcer, delayed healing, aphtha of mouth and tongue, gingival swelling and pain, laryngopharynx ulcer, and conjunctival congestion and pain caused by liver meridian wind heat or liver fire attack.
The traditional Chinese medicine components in the formula of the invention have the effects of nourishing kidney yin, clearing liver heat, nourishing liver and improving eyesight and the like, can complement each other, and can achieve the purposes of less feeding and high utilization rate of effective components by combining the preparation method of the invention.
Compared with the prior art, the invention has the following beneficial effects:
(1) the eye drop prepared by the invention is rich in polypeptides and various animal and plant active ingredients of traditional Chinese medicines, and the extracted traditional Chinese medicine extract and the active polypeptides have small molecular weight and can permeate various ocular tissue barriers (blood-retina inner and outer barriers and blood-aqueous humor barriers) to reach the bottom of eyes.
(2) The preparation process of the invention adopts the modern biological high-tech integration technology, and the effective components and active substances in the traditional Chinese medicine and animal tissues are retained to the maximum extent, for example, in the preparation process of the traditional Chinese medicine extract, the traditional Chinese medicine components are subjected to pretreatment such as dipping, ultrasonic treatment and the like at a specific temperature, so that the purification rate of the effective components of the traditional Chinese medicine is greatly improved; in the extraction process of the small molecular peptide, precipitation, ultrafiltration and nanofiltration multi-gradient concentration operations are performed, so that the extraction rate of the effective active ingredients of the traditional Chinese medicine extract and the small molecular peptide in the invention is ensured, and the production raw material cost is further reduced. Meanwhile, the active substances have synergistic effect, can supplement nutrition for eye nerve cells, and can nourish the visual nervous system and optic nerve cells; is suitable for treating eye diseases such as nebula, dry eye, asthenopia, blurred vision, and nyctalopia.
(3) The invention avoids the defects of the current clinical administration mode and has the advantages of simple and convenient administration process, no pain, no risk, furthest reduced treatment cost and the like.
(4) The eye drop rich in animal and plant active ingredients prepared by the invention can be prepared into various dosage forms such as different injections, aqueous solutions, atomizing agents, powders, gels, ointments, liquid dressings, eye patches, freeze-dried powder types and other administration modes such as administration devices or apparatuses by freeze-drying, spray-drying or excipient adding and the like, and can provide diversified products for people.
(5) The eye drop disclosed by the invention is safe, convenient and efficient, has no toxic or side effect, is easy to produce in a large scale, and can meet the needs of people, and the eye drop rich in animal and plant active ingredients, which is prepared by the invention, has the advantages that the peptide structure of animal peptides close to the human body is easy to absorb by the human body, the targeting effect is realized, the multifunctional advantage of plant peptides is realized, and the defect of a single kind of peptides is effectively overcome. Meanwhile, under the targeting effect of the animal peptide, the plant peptide is induced to act on eye nerve cells, and various small molecular peptides effectively play respective functions so as to improve the nourishing effect of the liquid medicine.
Detailed Description
The above-mentioned aspects of the present invention will be further described in detail with reference to the following specific examples. It should not be understood that the scope of the above-described subject matter of the present invention is limited to the following examples. Various substitutions and alterations according to the general knowledge and conventional practice in the art are intended to be included within the scope of the present invention without departing from the technical spirit of the present invention as described above.
Example 1
The eye drop rich in animal and plant active ingredients is characterized by comprising the following components in parts by weight: 0.1 part of active polypeptide, 5 parts of traditional Chinese medicine extract, 0.1 part of lubricating and moisturizing enhancer, 0.1 part of preservative, 1 part of osmotic pressure regulator, 0.5mg/mL of insulin, and the balance of physiological saline, wherein the pH is 6.0.
The active polypeptide is obtained by the following extraction process:
(1) taking healthy fresh animal tissues, tissue fluid or body fluid rich in active polypeptide, removing impurities, cleaning, cutting, and mashing with a tissue mashing machine;
(2) adding sodium chloride solution with equal mass, mixing, and grinding with colloid mill to obtain homogenate;
(3) adding 1 time of sodium chloride solution of equal mass, mixing, adding protease (including pancreatin, papain, and complex enzyme) 1 wt%, heating in water bath to 50 deg.C, adjusting pH to 7.5 with saturated calcium hydroxide, stirring at constant speed, and hydrolyzing for 10 hr;
(4) standing at-10 deg.C for 8 hr, and precipitating by cold precipitation.
(5) Taking out, centrifuging at 4000r/min for 5min at 0 ℃ to remove protein precipitate, and collecting supernatant:
(6) filtering the supernatant through a filter material with the aperture of 0.22um to remove impurities and micro particles, and collecting a clarified liquid:
(7) ultrafiltering, nanofiltering, and concentrating at multiple gradient to obtain active small molecular polypeptide with active molecular weight below 1000Da, and freezing for storage;
the traditional Chinese medicine extract is obtained by the following extraction process:
(1) weighing 40g of dendrobium, 120g of cassia seed, 50g of medlar, 130g of wild chrysanthemum flower and 10g of pearl in turn according to the requirements in the formula; slicing, cutting, pulverizing, placing in an ultrasonic reactor, stirring and mixing;
(2) adding 1.5 times volume of pure water or deionized water, soaking for 0.5 hr, and mixing to obtain mixed medicinal liquid;
(3) treating the mixed liquid medicine obtained in the step (2) for 5min by ultrasonic wave with the frequency of 10KHz and the temperature kept at 50 ℃, filtering by using a 100-mesh filter screen or gauze, collecting precipitates for treatment, and collecting filtrate for later use;
(4) putting the precipitate obtained in the step (3) into an ultrasonic reactor again, adding pure water or deionized water with the volume of 1.5 times, performing ultrasonic treatment for 5min at the ultrasonic frequency of 10kHz and the temperature of 50 ℃, filtering by using a 100-mesh filter screen or gauze with 150 meshes, and collecting the filtrate for later use;
(5) mixing the filtrates collected in steps (3) and (4) to obtain Chinese medicinal extract, and refrigerating for use.
The preparation method of the eye drop rich in animal and plant active ingredients in the embodiment comprises the following steps:
(1) preparation of a NaCl solution: weighing 0.5g of NaCl, adding deionized water to a constant volume of 100mL to obtain a NaCl solution;
(2) adding 0.1 part of active polypeptide and 5 parts of Chinese medicinal extract into 100mL of NaCl solution, dissolving completely, adjusting pH to 6.0, standing at 0 deg.C for 4 hr to obtain mixed solution of active small molecular polypeptide and Chinese medicinal extract;
(3) weighing 0.5g of sodium hyaluronate, 5g of polyoxyethylene 9-lauryl ether, 5g of polyoxyethylene 20-stearyl ether and 0.5g of insulin; sequentially adding the mixture into 500mL of pure water or deionized water, and stirring the mixture to dissolve the mixture to obtain a lubricating and moisturizing reinforcing agent solution;
(4) heating 100mL of purified water or deionized water to 40 ℃, adding 1 part of boric acid, stirring for dissolving, and standing at room temperature to obtain an osmotic pressure regulator;
(5) weighing 1 part of borneol, grinding into powder, adding 100mL of NaCl solution, and uniformly mixing to obtain a preservative;
(6) mixing the active small molecular polypeptide prepared in the steps (2), (3), (4) and (5) with the mixed solution of the traditional Chinese medicine extract, the lubrication and moisture retention enhancer, the preservative and the osmotic pressure regulator, uniformly mixing, supplementing physiological saline to 1000mL, adjusting the pH to 6.0, standing at 0-4 ℃ for 24h, and preparing into basic ophthalmic medicine;
(7) filtering and sterilizing by using filter materials with the pore diameters of 1.0um, 0.45um and 0.15um in a grading manner under an aseptic clean environment; sterile subpackaging to obtain the eye drop rich in animal and plant active ingredients.
The eye drop rich in animal and plant active ingredients prepared in this embodiment can be prepared into other dosage forms such as different injections, aqueous solutions, nebulizing agents, powders, gels, ointments, liquid dressings, eye patches, lyophilized powders and other administration modes such as administration devices or apparatuses by freeze-drying, spray-drying or adding excipients and other methods.
Example 2
An eye drop rich in animal and plant active ingredients comprises the following components in parts by weight: 5 parts of active polypeptide, 15 parts of traditional Chinese medicine extract, 5 parts of lubricating and moisturizing enhancer, 0.5 part of preservative, 5 parts of osmotic pressure regulator, and the balance of physiological saline for complement, wherein the pH value is 8.0.
The active polypeptide is obtained by the following extraction process:
(1) taking healthy fresh animal tissues, tissue fluid or body fluid rich in active polypeptide, removing impurities, cleaning, cutting, and mashing with a tissue mashing machine;
(2) adding sodium chloride solution with equal mass, mixing, and grinding with colloid mill to obtain homogenate;
(3) adding 3 times of sodium chloride solution, mixing, adding 2 wt% protease (including papain and complex enzyme), heating in water bath to 55 deg.C, adjusting pH to 8.5 with saturated calcium hydroxide, stirring at constant speed, and hydrolyzing for 15 hr;
(4) standing at 10 deg.C for 12h, and precipitating by isoelectric point precipitation;
(5) taking out, centrifuging at 4 ℃ at 10000r/min for 30min to remove protein precipitate, and collecting supernatant:
(6) filtering the supernatant through a filter material with the aperture of 1.2um to remove impurities and micro particles, and collecting a clarified liquid:
(7) ultrafiltering, nanofiltering and concentrating the collected clarified liquid in multiple gradients to obtain active small molecular polypeptide, and freezing and storing for later use;
the traditional Chinese medicine extract is obtained by the following extraction process:
(1) weighing 80g of dendrobium, 160g of cassia seed, 200g of medlar, 160g of wild chrysanthemum flower and 25g of pearl in turn according to the requirements in the formula; slicing, cutting, pulverizing, placing in an ultrasonic reactor, stirring and mixing;
(2) adding 3 times of pure water or deionized water, soaking for 2 hr, and mixing to obtain mixed medicinal liquid;
(3) treating the mixed liquid medicine obtained in the step (2) for 30min by ultrasonic wave at 200KHz frequency and at 60 ℃, filtering by using a 150-mesh filter screen or gauze, collecting precipitates for treatment, and collecting filtrate for later use;
(4) putting the precipitate obtained in the step (3) into an ultrasonic reactor again, adding 3 times of pure water or deionized water, performing ultrasonic treatment for 30min at the ultrasonic frequency of 200kHz and the temperature of 60 ℃, filtering by using a 150-mesh filter screen or gauze, and collecting filtrate for later use;
(5) mixing the filtrates collected in steps (3) and (4) to obtain Chinese medicinal extract, and refrigerating for use.
The preparation method of the eye drop rich in animal and plant active ingredients in the embodiment comprises the following steps:
(1) preparation of a NaCl solution: weighing 1.5g of NaCl, adding deionized water, and fixing the volume to 100mL to obtain a NaCl solution;
(2) adding 5 parts of active polypeptide and 15 parts of traditional Chinese medicine extract into 100mL of NaCl solution, fully dissolving, adjusting the pH value to 8.5, and standing at 8 ℃ for 8h to obtain a mixed solution of active small molecular polypeptide and traditional Chinese medicine extract;
(3) weighing 50g of sodium hyaluronate and 50g of polyoxyethylene 20-stearyl ether; sequentially adding the mixture into 500mL of pure water or deionized water, and stirring the mixture to dissolve the mixture to obtain a lubricating and moisturizing reinforcing agent solution;
(4) heating 100mL of purified water or deionized water to 50 ℃, adding 5 parts of NaCl, stirring for dissolving, and standing at room temperature to obtain an osmotic pressure regulator;
(5) weighing 10 parts of polylysine, adding the polylysine into 100mL of NaCl solution, and uniformly mixing to obtain the preservative;
(6) mixing the active small molecular polypeptide prepared in the steps (2), (3), (4) and (5) with the mixed solution of the traditional Chinese medicine extract, the lubrication and moisture retention enhancer, the preservative and the osmotic pressure regulator, uniformly mixing, supplementing physiological saline to 1000mL, adjusting the pH to 8.0, standing at 0-4 ℃ for 48h, and preparing into basic ophthalmic medicine;
(7) filtering and sterilizing by using filter materials with the pore diameters of 1.0um, 0.45um and 0.15um in a grading manner under an aseptic clean environment; sterile subpackaging to obtain the eye drop rich in animal and plant active ingredients.
The eye drop rich in animal and plant active ingredients prepared in this embodiment can be prepared into other dosage forms such as different injections, aqueous solutions, nebulizing agents, powders, gels, ointments, liquid dressings, eye patches, lyophilized powders and other administration modes such as administration devices or apparatuses by freeze-drying, spray-drying or adding excipients and other methods.
Example 3
An eye drop rich in animal and plant active ingredients comprises the following components in parts by weight: 3 parts of active polypeptide, 10 parts of traditional Chinese medicine extract, 3 parts of lubricating and moisturizing enhancer, 0.3 part of preservative, 3 parts of osmotic pressure regulator, and the balance of physiological saline, wherein the pH is 7.0.
The active polypeptide is obtained by the following extraction process:
(1) taking healthy fresh animal tissues, tissue fluid or body fluid rich in active polypeptide, removing impurities, cleaning, cutting, and mashing with a tissue mashing machine;
(2) adding sodium chloride solution with equal mass, mixing, and grinding with colloid mill to obtain homogenate;
(3) adding 2 times of sodium chloride solution of equal mass, mixing, adding compound protease 1.5% of the raw material weight, heating in water bath to 52 deg.C, adjusting pH to 8.0 with saturated calcium hydroxide, stirring at constant speed, and hydrolyzing for 12 hr;
(4) standing at 0 deg.C for 10h, wherein the precipitation method includes but is not limited to cold precipitation, salting-out precipitation, organic solvent precipitation or isoelectric point precipitation;
(5) taking out, centrifuging at 7000r/min at 2 ℃ for 20min to remove protein precipitate, and collecting supernatant:
(6) filtering the supernatant through a filter material with the aperture of 0.22um to remove impurities and micro particles, and collecting a clarified liquid:
(7) ultrafiltering, nanofiltering and concentrating the collected clarified liquid in multiple gradients to obtain active small molecular polypeptide, and freezing and storing for later use;
the traditional Chinese medicine extract is obtained by the following extraction process:
(1) weighing 60g of dendrobium, 150g of cassia seed, 100g of medlar, 150g of wild chrysanthemum flower and 15g of pearl in turn according to the requirements in the formula; slicing, cutting, pulverizing, placing in an ultrasonic reactor, stirring and mixing;
(2) adding 2 times of pure water or deionized water, soaking for 1 hr, and mixing to obtain mixed medicinal liquid;
(3) treating the mixed liquid medicine obtained in the step (2) for 20min by ultrasonic wave at the frequency of 100KHz and at the temperature of 55 ℃, filtering by using a 120-mesh filter screen or gauze, collecting precipitates to be treated, and collecting filtrate for later use;
(4) putting the precipitate obtained in the step (3) into an ultrasonic reactor again, adding 2 times of pure water or deionized water, performing ultrasonic treatment for 20min at an ultrasonic frequency of 100kHz and a temperature of 55 ℃, filtering with a 120-mesh filter screen or gauze, and collecting filtrate for later use;
(5) mixing the filtrates collected in steps (3) and (4) to obtain Chinese medicinal extract, and refrigerating for use.
The preparation method of the eye drop rich in animal and plant active ingredients in the embodiment comprises the following steps:
(1) preparation of a NaCl solution: weighing 1.0g of NaCl, adding deionized water, and fixing the volume to 100mL to obtain a NaCl solution;
(2) adding 3 parts of active small molecular polypeptide and 10 parts of traditional Chinese medicine extract into 100mL of NaCl solution, fully dissolving, adjusting the pH value to 7.5, and standing at 8 ℃ for 6h to obtain a mixed solution of the active polypeptide and the traditional Chinese medicine extract;
(3) weighing 30g of polyvinyl alcohol, 20g of carbomer sodium hyaluronate and 30g of sodium carboxymethylcellulose; sequentially adding the mixture into 500mL of pure water or deionized water, and stirring the mixture to dissolve the mixture to obtain a lubricating and moisturizing reinforcing agent solution;
(4) heating 100mL of purified water or deionized water to 45 ℃, adding 3 parts of boric acid, stirring for dissolving, and standing at room temperature to obtain an osmotic pressure regulator;
(5) weighing 5 parts of menthol, grinding into powder, adding 100mL of NaCl solution, and mixing uniformly to obtain a preservative;
(6) mixing the active small molecular polypeptide prepared in the steps (2), (3), (4) and (5) with the mixed solution of the traditional Chinese medicine extract, the lubrication and moisture retention enhancer, the preservative and the osmotic pressure regulator, uniformly mixing, supplementing physiological saline to 1000mL, adjusting the pH to 7.0, standing at 0-4 ℃ for 36h, and preparing into basic ophthalmic medicine;
(7) filtering and sterilizing by using filter materials with the pore diameters of 1.0um, 0.45um and 0.15um in a grading manner under an aseptic clean environment; sterile subpackaging to obtain the eye drop rich in animal and plant active ingredients.
The eye drop rich in animal and plant active ingredients prepared in this embodiment can be prepared into other dosage forms such as different injections, aqueous solutions, nebulizing agents, powders, gels, ointments, liquid dressings, eye patches, lyophilized powders and other administration modes such as administration devices or apparatuses by freeze-drying, spray-drying or adding excipients and other methods.
The above-mentioned embodiments are merely preferred embodiments of the present invention, which are not intended to limit the scope of the present invention, and therefore, all equivalent changes made by the contents of the claims of the present invention should be included in the claims of the present invention.
Claims (9)
1. The eye drop rich in animal and plant active ingredients is characterized by comprising the following raw materials in parts by weight: active small molecular polypeptide 0.1-5 parts, Chinese medicinal extract 5-15 parts, lubricant and moisture enhancer 0.1-5 parts, antiseptic 0.1-0.5 part, osmotic pressure regulator 1-5 parts, and physiological saline for balance, and pH is 6.0-8.0.
2. The eye drop rich in animal and plant active ingredients of claim 1, wherein: the eye drop also comprises insulin, and the addition amount of the insulin in the eye drop is 0.3-0.5 mg/mL.
3. The eye drop rich in animal and plant active ingredients of claim 1, wherein the lubricant moisturizing enhancer comprises one or more of sodium hyaluronate, polyvinyl alcohol, tween 80, hydroxyethyl cellulose, sodium carboxymethyl cellulose, carbomer, polyoxyethylene 9-lauryl ether, polyoxyethylene 20-stearyl ether, and glycerin.
4. The eye drop rich in animal and plant active ingredients as claimed in claim 1, wherein the preservative comprises one or more of borneol, menthol, polylysine, chitosan, chlorobutanol and borax.
5. An eye drop rich in animal and plant active ingredients according to claim 1, wherein the osmotic pressure regulator comprises one or more of sodium chloride, boric acid, potassium nitrate, glucose, etc.
6. The preparation method of the eye drop rich in animal and plant active ingredients, which is characterized by comprising the following steps:
(1) preparation of a NaCl solution: weighing 0.5-1.5g of NaCl, adding deionized water to a constant volume of 100mL to obtain a NaCl solution;
(2) adding 0.1-5 parts of active small molecular polypeptide and 5-15 parts of Chinese medicinal extract into 100mL of NaCl solution, dissolving completely, adjusting pH to 6.0-8.5, and standing at 0-8 deg.C for 4-8 hr to obtain mixed solution of active small molecular polypeptide and Chinese medicinal extract;
(3) weighing 0.1-5 parts of lubricating and moisturizing reinforcing agent; sequentially adding the mixture into 500mL of pure water or deionized water, and stirring the mixture to dissolve the mixture to obtain a lubricating and moisturizing reinforcing agent solution;
(4) heating 100mL of purified water or deionized water to 40-50 ℃, adding 1-5 parts of osmotic pressure regulator, stirring for dissolving, and standing at room temperature to obtain the osmotic pressure regulator;
(5) weighing 1-10 parts of antiseptic materials such as borneol and the like, grinding into powder, and adding the powder into 100mL of NaCl solution for uniformly mixing to obtain an antiseptic;
(6) mixing the active small molecular polypeptide prepared in the steps (2), (3), (4) and (5) with the mixed solution of the traditional Chinese medicine extract, the lubrication and moisture retention enhancer, the preservative and the osmotic pressure regulator, uniformly mixing, supplementing physiological saline to 1000mL, adjusting the pH to 6.0-8.0, standing at 0-4 ℃ for 24-48h, and preparing into basic ophthalmic medicine;
(7) filtering and sterilizing by using filter materials with the pore diameters of 1.0um, 0.45um and 0.15um in a grading manner under an aseptic clean environment; sterile subpackaging to obtain the eye drop rich in animal and plant active ingredients.
7. The eye drop rich in animal and plant active ingredients of claim 1, wherein the extraction process of the active polypeptide comprises the following steps:
(1) crushing raw materials: taking healthy fresh animal tissues, tissue fluid or body fluid rich in active polypeptide, removing impurities, cleaning, cutting, and mashing with a tissue mashing machine;
(2) grinding: adding sodium chloride solution with equal mass into the mashed raw materials, uniformly mixing, and grinding into a homogenate;
(3) enzyme digestion: adding 1-3 times of sodium chloride solution into the homogenate obtained in the step (2), mixing uniformly, and adding protease 1-2% of the weight of the raw material, wherein the protease includes but is not limited to pancreatin, papain and complex enzyme; heating the mixture in water bath to 50-55 deg.C, adjusting pH to 7.5-8.5 with saturated calcium hydroxide, stirring at constant speed, and hydrolyzing for 10-15 hr;
(4) and (3) precipitation: standing at-10-10 deg.C for 8-12h, wherein the precipitation method includes but is not limited to cold precipitation, isoelectric point precipitation, organic solvent precipitation or salting-out precipitation;
(5) centrifuging: taking out, centrifuging at 4000 and 10000r/min at 0-4 ℃ for 5-30min to remove protein precipitate, and collecting supernatant:
(6) and (3) filtering: filtering the supernatant through a filter material with the aperture of 0.22-1.2um to remove impurities and micro particles, and collecting a clarified liquid:
(7) concentration: ultrafiltering, nanofiltering, concentrating with multi-gradient to obtain active small molecular polypeptide with molecular weight below 1000Da, and freezing for storage.
8. The eye drop rich in animal and plant active ingredients of claim 1, wherein the extraction process of the traditional Chinese medicine extract comprises the following steps:
(1) weighing 40-80g of dendrobium, 120-160g of cassia seed, 50-200g of medlar, 130-160g of wild chrysanthemum and 10-25g of pearl in turn; slicing, cutting, pulverizing, placing in an ultrasonic reactor, stirring and mixing;
(2) adding 1.5-3 times of pure water or deionized water, soaking for 0.5-2 hr, and mixing to obtain mixed medicinal liquid;
(3) treating the mixed liquid medicine obtained in the step (2) for 5-30min by ultrasonic wave with the frequency of 10-200KHz and the temperature kept at 50-60 ℃, filtering by using a 100-mesh filter screen or gauze, collecting precipitates to be treated, and collecting filtrate for later use;
(4) putting the precipitate in the step (3) into an ultrasonic reactor again, adding 1.5-3 times of pure water or deionized water, performing ultrasonic treatment for 5-30min at the ultrasonic frequency of 10-200kHz and the temperature of 50-60 ℃, filtering by using a 100-mesh filter screen or gauze, and collecting the filtrate for later use;
(5) mixing the filtrates collected in steps (3) and (4) to obtain Chinese medicinal extract, and refrigerating for use.
9. The eye drop rich in animal and plant active ingredients of claim 1, wherein the eye drop rich in animal and plant active ingredients prepared by the invention can be prepared into different injections, aqueous solutions, atomizing agents, powders, gels, ointments, liquid dressings, eye patches and freeze-dried powders by freeze-drying, spray-drying or excipient adding methods.
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