WO2010083636A1 - 小牛血去蛋白提取物的滴眼液 - Google Patents

小牛血去蛋白提取物的滴眼液 Download PDF

Info

Publication number
WO2010083636A1
WO2010083636A1 PCT/CN2009/001475 CN2009001475W WO2010083636A1 WO 2010083636 A1 WO2010083636 A1 WO 2010083636A1 CN 2009001475 W CN2009001475 W CN 2009001475W WO 2010083636 A1 WO2010083636 A1 WO 2010083636A1
Authority
WO
WIPO (PCT)
Prior art keywords
weight
parts
calf blood
ophthalmic solution
eye
Prior art date
Application number
PCT/CN2009/001475
Other languages
English (en)
French (fr)
Inventor
刘继东
艾立诚
杨宇春
Original Assignee
沈阳兴齐制药有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 沈阳兴齐制药有限公司 filed Critical 沈阳兴齐制药有限公司
Priority to US13/145,311 priority Critical patent/US20120003323A1/en
Priority to EP09838603.0A priority patent/EP2389924B1/en
Publication of WO2010083636A1 publication Critical patent/WO2010083636A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • 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
    • A61K9/0048—Eye, e.g. artificial tears
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/075—Ethers or acetals
    • A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A61K31/722—Chitin, chitosan
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A—HUMAN NECESSITIES
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents

Definitions

  • the present invention relates to an eye drop containing a deproteinized extract of calf blood. Background technique:
  • the calf blood deproteinized extract is a biological preparation invented by the German scholar Professor Jaeger. It is made from the blood of healthy and well-developed calves within six months of the newborn. The molecular weight of the membrane is less than 6000.
  • the deproteinized blood extract contains an electrolyte, a trace amount of essential elements, and an intermediate product of a nucleic acid derivative, an amino acid, an oligosaccharide, a low molecular polypeptide, a glycolipid, and a lipid metabolism.
  • Calf blood deproteinized extract promotes the uptake and utilization of glucose and oxygen by the mitochondria at the cellular level (independent of insulin), increases ATP synthesis, increases cellular energy, and activates multiple enzyme activities. In the case of hypoxemia and increased energy requirements, it rapidly improves the hypoxic state of tissue cells, improves microcirculation, activates tissue cell proliferation, and promotes repair of damaged tissue cells. It is a highly potent cytotrophic factor and cell growth factor.
  • the calf blood deproteinized extract contains no antigen, is well tolerated, safe and non-toxic. As a drug that promotes cell energy metabolism, the clinical application effect is good. Calf blood deproteinized ophthalmic liquid preparation can improve local blood circulation and dystrophies by improving the uptake and utilization of oxygen and glucose by tissue cells, and can also promote the migration and proliferation of sputum cells and vascular endothelial cells. Role, clinically used for the treatment of traumatic, inflammatory and nutritional corneal, conjunctival lesions and dry eye.
  • New drug products for the treatment of such diseases are: recombinant bovine basic fibroblast growth factor, recombinant human epidermal growth factor, all genetically engineered products, although theoretically promote epidermal growth, but with actual surface growth
  • the mediums needed in the process are far apart and cannot be compared with the calf blood extract.
  • Ophthalmic ointment for calf blood deproteinized extract has been marketed abroad.
  • the product produced in Switzerland under the trade name "Su Gao Jie” is an ophthalmic ointment with calf blood deproteinized extract as an active ingredient, which has obvious curative effect on wound healing of ocular tissues.
  • Protein extract Ophthalmic preparations, especially eye drops, have not been used clinically in China.
  • the present inventors have found that the use of chitosan oligosaccharides can effectively improve the stability of deproteinized extracts of calf blood, and the present inventors have achieved the present invention based on this finding.
  • the present invention provides an ophthalmic eye drop comprising a therapeutically effective amount of calf blood deproteinized extract, chitooligosaccharide, and water.
  • the ophthalmic solution of the present invention comprises 10 to 30% (v/v) of calf blood deproteinized extract, 0.01 to 1% (w/v) based on the total amount of the ophthalmic solution. ) Chitosan oligosaccharides, and water.
  • the chitooligosaccharide has a molecular weight of less than 5,000, preferably less than 3,000, preferably less than 2,000, preferably less than 1,500.
  • it further comprises one or more bacteriostatic agents selected from the group consisting of: phenylethyl alcohol, phenoxyethanol, decyl hydroxybenzoate, ethyl p-hydroxybenzoate , propyl paraben, benzalkonium chloride, chlorobutanol.
  • the bacteriostatic agent is one or more selected from the group consisting of phenoxyethanol, ethyl p-hydroxybenzoate, benzalkonium chloride, and chlorobutanol. In still further embodiments, the bacteriostatic agent is a combination of phenoxyethanol and ethyl p-hydroxybenzoate. In still further embodiments, the bacteriostatic agent accounts for 0.005 to 1% of the total amount of the eye drops (w/v is preferred, and the bacteriostatic agent accounts for 0.01-0.5 of the total amount of the eye drops) % (w/v). Preferably, the bacteriostatic agent accounts for 0.1-0.5% (w/v) of the total amount of the eye drops. Preferably, the bacteriostatic agent accounts for the total amount of the eye drops. 0.01-0.1% (w/v preferably, the bacteriostatic agent accounts for 0.01, 0.1, or 0.5% (w/v) of the total amount of the eye drops.
  • the ophthalmic solution of the present invention further comprises a pH adjusting agent.
  • the eye drops have a pH of from 6.0 to 9.0, preferably from 6.0 to 8.5, preferably from 6.0 to 8.0.
  • the pH adjusting agent is selected from the group consisting of phosphates, acetates, borates, and combinations thereof, such as a combination of sodium dihydrogen phosphate and disodium hydrogen phosphate, a combination of sodium acetate and acetic acid, boric acid.
  • a combination of sodium and boric acid, etc. can be determined by those skilled in the art according to the specific conditions.
  • the ophthalmic solution of the present invention further comprises an osmotic pressure regulating agent.
  • the osmotic pressure adjusting agent include sodium chloride, boric acid, potassium nitrate, glucose, and the like, and those skilled in the art can determine the amount thereof according to a specific case.
  • the ophthalmic solution of the present invention further comprises a viscosity modifier, which is also generally understood by those skilled in the art as a thickener.
  • a viscosity modifier which is also generally understood by those skilled in the art as a thickener.
  • the viscosity modifier of the present invention include polyvinylpyrrolidone, sodium carboxymethylcellulose, decylcellulose, polyvinyl alcohol, etc., and the amount thereof can be determined by a person skilled in the art according to a specific case.
  • Various aspects of the invention and their benefits are further described below. All documents cited in the present invention are hereby incorporated by reference in their entirety, and if the meanings expressed by these documents are inconsistent with the present invention, the expression of the present invention shall prevail.
  • v / v and "w / v” means the percentage of each "v / v" (i.e. ml / 100ml) and a percent "by weight / volume" (i.e. g / 100ml) o
  • the calf blood deproteinized extract of the present invention is obtained by a process in which blood is removed from the calf, which is less than 6 months after birth, and the blood is removed by protein removal, virus inactivation, and ultrafiltration.
  • the calf blood deproteinized extract should meet the "Buffalo Blood Deproteinized Extract Quality Standard" (this standard is the standard approved by the State Food and Drug Administration, the standard number is YBH 05782007, and the approval time is December 29, 2007. ) requirements.
  • the extract is made by deproteinization, concentration, ultrafiltration and other processes of the calf blood, and contains active ingredients such as amino acids and small molecule peptides.
  • the total solid content should be not less than 35m g /ml.
  • the more detailed preparation method and/or quality index of the calf blood deproteinized extract of the present invention is further illustrated as follows:
  • the raw material calf blood deproteinized extract used in the ophthalmic solution of the present invention can be used from the Songnen Plain pasture in Heilongjiang province.
  • the fetal calf, this pasture is one of the three high-quality pastures in the world.
  • the calf blood of the northern yellow cattle species with a quarantine certificate of not more than 6 months after birth is used as raw material. Under decontamination conditions, fibrin is removed and concentrated. And the membrane separation technology is used for ultrafiltration to obtain the calf blood deproteinized extract of the desired molecular weight.
  • the extract contains active ingredients such as amino acids and low molecular peptides, and can be used as an ophthalmic gel and drops.
  • the raw material of eye drops is mainly used for the treatment of traumatic, inflammatory and nutritional corneal and conjunctival lesions.
  • Specific preparation process package Included: (1) Calf blood protein removal: The defibrinated calf blood after freezing and thawing is added to the extractant in a certain proportion under aseptic conditions, stirred for a specified time, and allowed to stand under the conditions of the specified temperature and humidity.
  • the ophthalmic preparation prepared from the calf blood deproteinized extract should be controlled to have a viability of not less than 4.0 ⁇ l ⁇ 2 / ⁇ ⁇ .1 ⁇ , and the polypeptide per liter of eye drops should be 0.18 mg - 0.28 mg.
  • Chitosan Oligosaccharide ( ⁇ -1,4-oligo-D-glucosamine, also known as chitosan oligosaccharide, amino oligosaccharide) of the present invention, which refers to 2 to 10 glucosamines as ⁇ -1
  • An oligosaccharide linked by a 4-glycosidic bond which is usually obtained by degradation of chitosan, a product of deacetylation of chitin (shrimp, crab shell, etc.).
  • Chitosan oligosaccharides can be obtained by known methods or directly by commercial purchase, for example, from the company of Kelianke Biotechnology Co., Ltd.
  • the chitosan oligosaccharides of different molecular weights may be used as needed.
  • the chitosan oligosaccharide has a molecular weight of less than 5,000.
  • the chitooligosaccharide has a molecular weight of less than 3,000.
  • the chitooligosaccharide has a molecular weight of less than 2,000.
  • the chitooligosaccharide has a molecular weight of less than 1500.
  • the ophthalmic solution of the present invention contains 0.01 to 1% (w/v) of chitosan oligosaccharide. In another embodiment of the ophthalmic solution of the present invention, it contains 0.01 to 0.5% (w/v) of chitosan oligosaccharide. In still another embodiment of the ophthalmic solution of the present invention, it contains 0.05 to 0.2% (w/v) of chitosan oligosaccharide. In still another embodiment of the ophthalmic solution of the present invention, it contains 0.05 or 0.1% (w/v) of chitosan oligosaccharide.
  • the ophthalmic solution of the present invention is an aqueous composition in which water is used as a barrier or vehicle or excipient or carrier for the composition.
  • the ophthalmic solution of the present invention does not specifically indicate the percentage of water used as a percentage of the total weight of the ophthalmic solution, or the amount of water used is not specifically indicated in the preparation of the ophthalmic solution of the present invention, it will be apparent to those skilled in the art that As the medium shield or vehicle or vehicle or carrier of the eye drops, the amount of water is calculated by adding water to the total amount of the eye drops, which is generally used in the preparation of liquid preparations such as eye drops and injections. Measurement method.
  • the bacteriostatic agent may be in any combination of one or more.
  • the bacteriostatic agent is a combination of phenoxyethanol and ethyl p-hydroxybenzoate; in a further embodiment, the bacteriostatic agent is phenoxyethanol and p-hydroxybenzoic acid.
  • the ethyl ester is combined in any ratio; in still further embodiments, the bacteriostatic agent is a combination of phenoxyethanol and ethyl p-hydroxybenzoate in a weight ratio of 50:1.
  • the bacteriostatic agent is a combination of phenoxyethanol and benzalkonium chloride; in a further embodiment, the bacteriostatic agent is phenoxyethanol and benzalkonium chloride in any ratio. Combination; In still further embodiments, the bacteriostatic agent is phenoxyethanol and benzalkonium chloride combined in a weight ratio of 50:1. In one embodiment, the bacteriostatic agent is a combination of chlorobutanol and benzalkonium chloride; in a further embodiment, the bacteriostatic agent is chlorobutanol and benzalkonium chloride.
  • the bacteriostatic agent is chlorobutanol and benzalkonium chloride combined in a weight ratio of 10:1.
  • the bacteriostatic agent is a combination of chlorobutanol and ethyl p-hydroxybenzoate; in a further embodiment, the bacteriostatic agent is chlorobutanol and p-hydroxyphenylhydrazine.
  • the acid ethyl ester is combined in any ratio; in still further embodiments, the bacteriostatic agent is a combination of chlorobutanol and ethyl p-hydroxybenzoate in a weight ratio of 10:1.
  • the second aspect of the present invention may further provide an eye drop, which comprises the eye drop according to any one of the first aspects of the present invention, which may further contain a pharmaceutically acceptable excipient, for example, the present invention may be
  • the eye drops of the first aspect are diluted with the appropriate solvent to the ready-to-use eye drops of the second aspect of the invention prior to clinical use to obtain a clinically useful effect, such as long-term preservation of the eye drops of the first aspect of the invention.
  • Applicable ring When the pH is inconsistent with the physiological pH, an appropriate amount of the pH adjuster may be added to the eye drops of the first aspect of the invention before use, so that the pH of the obtained eye drops of the second aspect of the invention is adjusted to the eye.
  • a physiologically acceptable pH environment may be added to the eye drops of the first aspect of the invention before use, so that the pH of the obtained eye drops of the second aspect of the invention is adjusted to the eye.
  • the invention further relates to eye drops having a formulation of an embodiment of the invention.
  • the invention further relates to eye drops having the following formulation:
  • the invention further relates to eye drops having the following formulation:
  • the invention further relates to eye drops having the following formulation:
  • the invention further relates to eye drops having the following formulation:
  • the third aspect of the present invention provides the ophthalmic solution according to any one of the first aspect of the present invention or the ophthalmic solution according to any one of the second aspect of the present invention, which is used for the treatment of an eye disease (for example, a corneal disease such as corneal damage, Use of medicine for conjunctival lesions, dry eye syndrome.
  • an eye disease for example, a corneal disease such as corneal damage, Use of medicine for conjunctival lesions, dry eye syndrome.
  • a fourth aspect of the present invention provides a method of treating and/or preventing an eye disease (for example, a corneal disease such as corneal damage, conjunctival lesion, dry eye), comprising administering a treatment and/or prevention to a subject in need thereof An effective amount of the eye drops of any one of the first aspects of the invention or the eye drops of any of the second aspect of the invention.
  • an eye disease for example, a corneal disease such as corneal damage, conjunctival lesion, dry eye
  • the term "subject” refers to, but is not limited to, mammals such as pigs, dogs, monkeys, cats, humans, etc., and birds such as chickens, ducks, etc., preferably, the subject is a human, in particular It is a person who may or may suffer from an eye disease as described herein (eg, corneal disease such as corneal damage, conjunctival lesions, dry eye).
  • eye disease eg, corneal disease such as corneal damage, conjunctival lesions, dry eye.
  • the ophthalmic solution of the present invention can be prepared by the following process: weighing a prescription amount of chitooligosaccharide under aseptic conditions (and, if necessary, a bacteriostatic agent such as a combination of phenoxyethanol and hydroxyethyl ester, pH adjustment Agent, osmotic pressure regulator, viscosity modifier, etc.), dissolve with appropriate amount of water for injection, add calf blood to protein extract, stir and hook, add water for injection to nearly full amount. First, filter with 0.45 ⁇ microporous membrane, add water for injection to the whole amount, and finally filter it with 0.22 ⁇ microporous membrane.
  • a bacteriostatic agent such as a combination of phenoxyethanol and hydroxyethyl ester, pH adjustment Agent, osmotic pressure regulator, viscosity modifier, etc.
  • the eye drops of the calf blood deproteinized extract prepared according to the present invention have a fixed source of calf blood and a stable quality, and the production process is easy to control.
  • the prepared calf blood deproteinized ophthalmic liquid preparation has a clear appearance, sterility, low irritation and low toxicity, and can provide a safe and effective ophthalmic preparation for clinical use.
  • the eye drops of the deproteinized extract of calf blood provided by the invention can promote the utilization of oxygen by the cells by increasing the respiration capacity of the mitochondria and the synthesis of high-energy phosphoric acid, and can maintain the physiological functions of important enzymes in the human body, therefore, it has Improve tissue nutrition and stimulation Cell regeneration and accelerated tissue repair, and can hyperplasia of hyperproliferative granulation tissue, collagen fiber reorganization, reduce or avoid scar formation.
  • the eye drops of calf blood deproteinized extract have good surface activity and good eye tolerance. They can be evenly distributed on the surface of the keratoconjunctiva, thereby lubricating the cornea, reducing the mechanical friction of the eyelid on the corneal surface and reducing the local irritation. Therefore, for corneal diseases, especially corneal damage, on the basis of conventional treatment, combined application of eye drops of calf blood deproteinized extract can promote the growth and repair of corneal epithelium and accelerate wound healing.
  • the eye drops of calf blood deproteinized extract provided by the invention can be used for treating corneal and conjunctival lesions, and can also be used for dry eye syndrome, for traumatic, inflammatory and nutritional cornea, conjunctival lesions and dry eye syndrome.
  • the treatment brings new options. detailed description:
  • the present invention provides a general and/or specific description of the materials and test methods used in the tests. While many of the materials and methods of operation used to accomplish the objectives of the present invention are well known in the art, the present invention is still described in detail herein. It will be apparent to those skilled in the art that, hereinafter, the materials and methods of operation of the present invention are well known in the art unless otherwise stated.
  • the total solid content of the calf blood deproteinized extract used in the following examples of the present invention is not less than 35 mg/ml.
  • the prescribed amount of phenoxyethanol, benzalkonium chloride, and chitosan oligosaccharide are heated and dissolved with an appropriate amount of water for injection, and cooled to room temperature, and the deproteinized extract of calf blood and water for injection are added to the whole amount. It was first filtered with a 0.45 ⁇ m ⁇ microporous membrane, and then sterilized by a 0.22 ⁇ microporous membrane filter, and filled to obtain an eye drop.
  • the chitosan oligosaccharide used in this example has a molecular weight of 5,000 or less.
  • the prescribed amount of phenoxyethanol, hydroxyphenylethyl ester and chitosan oligosaccharide were heated and dissolved with an appropriate amount of water for injection, and cooled to room temperature, and the calf blood deproteinized extract and water for injection were added to the whole amount. It was first filtered with a 0.45 ⁇ m ⁇ microporous membrane, and then sterilized by a 0.22 ⁇ m microporous membrane filter, and filled, and an eye drop was obtained.
  • the molecular weight of the chitooligosaccharide used in the present example was 3,000 or less.
  • Test drug The eye drop of the embodiment 1 of the present invention, the blank control solution of the eye drop, provided by the company.
  • Test drug The eye drops of Example 1 of the present invention, the blank control solution of the eye drops, and the positive sensitizer (2,4-dinitrochlorobenzene) were supplied by the company.
  • guinea pigs were randomly divided into 3 groups, 10 in each group, both male and female. The back sides of the guinea pigs were depilated 24 h before administration. Among them, group I was given the eye drops of the present invention; group II was given a blank control solution, and group m was given a positive sensitizer.
  • Sensitized exposure 0.2 ml of the ophthalmic liquid preparation was applied to the left hair removal area of the guinea pig for 6 hours. Days 7d and 14d are repeated in the same way.
  • the blank control solution and the positive sensitizer group method were the same as the ophthalmic liquid preparation group.
  • the calf blood deproteinized extract eye drops were applied to the guinea pig skin for sensitized contact and stimulating contact, no erythema and edema appeared, no difference from the blank control solution group, and the sensitization rate of the positive sensitizer group was 100%, indicating This product has no local sensitization of the skin.
  • Calf blood deproteinized extract ophthalmic liquid preparations were administered to guinea pigs without local sensitization. Stability example:
  • Test Drugs Eye drops of Examples 3 and 4 of the present invention and Comparative Examples 1 and 2 (referred to as Control 1 and Control 2, respectively) were supplied by the company.
  • pH determination Refer to the Chinese Pharmacopoeia 2005 edition two appendix VI ⁇ operation. The pH is between 6.0 and 9.0.
  • Visible foreign matter Refer to the Chinese Pharmacopoeia 2005 edition two appendix C ⁇ visible foreign bodies Check the inspection method.
  • Vitality measurement Take this product, use Vapor's breath tester to measure the respiratory vitality Q0 2 of guinea pig liver homogenate, and the respiratory vitality should be no less than 4.0 l O 2 /mg'h.
  • Peptide content Refer to the National Drug Standards, Book 16 16-119 Forin Phenol Determination, spectrophotometric determination.
  • the vitality measurement should satisfy Q0 2 ⁇ 4. It can be seen from the results that the protein viability still meets the requirements of the eye drops after 18 months compared with the absence of chitosan oligosaccharides. It indicates that chitosan oligosaccharides can protect the respiratory respiration of proteins to a certain extent, making the preparation more stable. Therefore, it was confirmed by the stability experiment results that the addition of chitosan oligosaccharide significantly improved the stability of the ophthalmic solution of the present invention.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Virology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Molecular Biology (AREA)
  • Inorganic Chemistry (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Cell Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines Containing Plant Substances (AREA)

Description

小牛血去蛋白提取物的滴眼液 本申请要求于 2009年 1月 20日提交的中国专利申请号 2009 1000 0893.9的优先权， 其全部内容通过引用并入本文。 技术领域：
本发明涉及一种含有小牛血去蛋白提取物的滴眼液。 背景技术：
小牛血去蛋白提取物是德国学者 Jaeger教授研究发明的一 种生物制剂，它是从新生六个月以内的健康且发育旺盛的小牛血 液中， 用膜过滤技术制成的分子量小于 6000的去蛋白血液提取 物。这种去蛋白质血液提取物中含有电解质、微量的必需元素以 及核酸衍生物、 氨基酸、 低聚糖、 低分子多肽、 糖脂类和类脂代 谢的中间产物等。小牛血去蛋白提取物能在细胞水平上促进细胞 线粒体对葡萄糖和氧的摄取和利用（不依赖胰岛素）， 使 ATP合 成增加， 细胞能量提高， 激活多种酶活力。 在低血氧以及能量需 增加等情况下， 迅速改善组织细胞乏氧状态， 改善微循环， 激活 组织细胞增生， 促进损伤组织细胞的修复，是一种高效细胞营养 素及促细胞生长因子。
小牛血去蛋白提取物不含抗原， 耐受良好， 安全无毒， 作为一 种促进细胞能量代谢的药物， 临床应用效果良好。 小牛血去蛋白提 取物眼用液体制剂通过改善组织细胞对氧和葡萄糖的摄取及利 用， 改善局部血液循环和营养障碍， 也能通过促进成仟维细胞和 血管内皮细胞的游走和增殖等作用， 临床上主要用于创伤性、 炎 症性和营养性角膜、 结膜病变及干眼症等的治疗。 目前， 已上市 的用于治疗此类疾病的新药产品有：重组牛碱性成纤维细胞生长因 子， 重组人表皮生长因子， 均为基因工程产物， 虽在理论上有促进 表皮生长作用， 但与实际体表生长中所需要的媒介相差甚远， 尚不 能和小牛血提取物相提并论。
小牛血去蛋白提取物的眼用膏剂已在国外上市。例如由瑞士生 产、 商品名为"素高捷疗，，的产品， 是以小牛血去蛋白提取物为活性 成分的眼膏剂， 其对眼组织的创伤修复有明显的疗效。 小牛血去蛋 白提取物眼用制剂特别是滴眼液尚未在国内用于临床。
尽管目前在国内有小牛血去蛋白提取物注射液在临床上使用， 但是本发明人发现， 这种加有吐温 -80的注射液因为稳定性不理想 而并不适合作为眼用制剂特别是作为滴眼液使用。 因此， 开发适合 临床应用的小牛血去蛋白提取物的眼用制剂特别是滴眼液，仍是本 领域技术人员急需解决的问题。 发明内容：
本发明的目的在于提供一种适合临床应用的含有小牛血去蛋 白提取物的滴眼液。 本发明人发现， 使用壳寡糖可以有效地提高 小牛血去蛋白提取物的稳定性， 本发明人基于该发现而得以实现 本发明。
为此， 本发明提供了一种供眼用的滴眼液， 其包含治疗有效 量的小牛血去蛋白提取物、 壳寡糖、 和水。
在本发明滴眼液的一个实施方案中， 按该滴眼液的总量计， 其包含 10~30%(v/v)的小牛血去蛋白提取物、 0.01〜l%(w/v)的壳寡 糖、 和水。
在本发明的一个实施方案中，所述壳寡糖的分子量小于 5000， 优选小于 3000， 优选小于 2000， 优选小于 1500。 在本发明滴眼液的一个实施方案中， 其还包含一种或多种选 自以下的抑菌剂： 苯乙醇、 苯氧乙醇、 对羟基苯曱酸曱酯、 对羟 基苯曱酸乙酯、 对羟基苯甲酸丙酯、 苯扎氯铵、 三氯叔丁醇。 在 进一步的实施方案中， 所述的抑菌剂是选自苯氧乙醇、 对羟基苯 甲酸乙酯、 苯扎氯铵、 三氯叔丁醇中的一种或多种。 在再进一步 的实施方案中， 所述的抑菌剂是苯氧乙醇和对羟基苯曱酸乙酯的 组合。 在更进一步的实施方案中， 所述的抑菌剂占该滴眼液总量 的 0.005~1% (w/v 优选的， 所述的抑菌剂占该滴眼液总量的 0.01-0.5% (w/v)。 优选的， 所述的抑菌剂占该滴眼液总量的 0.1-0.5% (w/v) . 优选的， 所述的抑菌剂占该滴眼液总量的 0.01-0.1% (w/v 优选的， 所述的抑菌剂占该滴眼液总量的 0.01、 0.1、 或 0.5% (w/v)„
在本发明滴眼液的一个实施方案中， 其还包含 pH调节剂。 在进一步的实施方案中，该滴眼液的 pH值为 6.0至 9.0，优选 6.0 至 8.5， 优选 6.0至 8.0。 在再在进一步的实施方案中， 所述的 pH 调节剂选自磷酸盐、 乙酸盐、 硼酸盐及其组合， 例如磷酸二氢钠 与磷酸氢二钠组合、 乙酸钠和乙酸组合、 硼酸钠和硼酸组合等， 本领域技术人员根据具体情况可以确定其用量。
在本发明滴眼液的一个实施方案中，其还包含渗透压调节剂。 所述的渗透压调节剂的实例包括氯化钠、 硼酸、 硝酸钾、 葡萄糖 等， 本领域技术人员根据具体情况可以确定其用量。
在本发明滴眼液的一个实施方案中， 其还包含粘度调节剂， 本领域技术人员通常还将这种粘度调节剂理解为增稠剂。 本发明 所述的粘度调节剂的实例包括聚乙烯吡咯烷酮、羧甲基纤维素钠、 曱基纤维素、 聚乙烯醇等， 本领域技术人员根据具体情况可以确 定其用量。 下面对本发明的各个方面以及它们的益处作进一步的描述。 本发明所引述的所有文献， 它们的全部内容通过引用并入本 文， 并且如果这些文献所表达的含义与本发明不一致时， 以本发 明的表述为准。 此外， 本发明使用的各种术语和短语具有本领域 技术人员公知的一般含义， 即便如此， 本发明仍然希望在此对这 些术语和短语作更详尽的说明和解释， 提及的术语和短语如有与 公知含义不一致的， 以本发明所表述的含义为准。
在本发明的上下文中， 用于表述。 /。的单位 "v/v"和 "w/v"分别 是指"体积 /体积"的百分数 (即 ml/100ml)和"重量 /体积"的百分数 (即 g/100ml)o
本发明的小牛血去蛋白提取物是通过从出生后不满 6个月的 小牛釆血， 血液经去除蛋白、 病毒灭活、 超滤截留等工艺获得。 该小牛血去蛋白提取物应当满足《小牛血去蛋白提取物质量标准》 (该标准是国家食品药品监督管理局所批复的标准， 标准号为 YBH 05782007, 批复时间为 2007年 12月 29日）的要求。 本提 取物是小牛血经过去蛋白、 浓缩、 超滤等工艺精制而成， 含有氨 基酸、 小分子肽等活性成分。 含总固体量应不少于 35mg/ml。
在本发明的小牛血去蛋白提取物的更详细的制备方法和 /或 质量指标进一步说明如下： 本发明滴眼液所使用的原材料小牛血 去蛋白提取物可以采用来自黑龙江松嫩平原牧场的胎牛， 此牧场 是世界上三大优质牧场之一， 附有检疫合格证的出生后不超过 6 个月的北方黄牛种的小牛血为原料， 在净化条件下， 去除纤维蛋 白， 浓缩， 并以膜分离技术超滤截留， 得到所需分子量的小牛血 去蛋白提取物， 此提取物经病毒灭活后， 含有氨基酸、 低分子肽 等活性成分， 可以作为眼用凝胶及滴眼液的原料， 主要用于创伤 性、 炎症性和营养性角膜、 结膜病变的治疗。 具体的制备工艺包 括： （1)小牛血除蛋白： 将冻融后的去纤维蛋白小牛血在无菌条件 下， 按一定比例加入提取剂， 搅拌至规定时间， 在工艺规定温湿 度条件下静置一定时间； （2)蒸发浓缩： 用旋转蒸发仪减压浓缩； (3)超滤截留： 将截留有效成分后的浓缩液， 经膜分离技术可得小 牛血去蛋白提取物。 由该小牛血去蛋白提取物配制的眼用制剂应 控制其活力测定不小于 4.0μ1 Ο2/ιηδ.1ι， 每亳升滴眼液含多肽应为 0.18mg - 0.28mg。
本发明的壳寡糖 (Chitosan Oligosaccharide, β-1，4-寡聚 -D-氨 基葡萄糖， 又称几丁寡糖、 氨基寡糖素)， 它是指 2 ~ 10个氨基葡 萄糖以 β-1，4-糖苷键连接而成的低聚糖， 它通常是由曱壳素 (虾、 蟹壳等)脱乙酰化的产物壳聚糖的降解而获得。 壳寡糖可以通过已 知的方法获得， 也可直接通过商业购买而获得， 例如可以购自大 连中科格莱克生物科技有限公司。 可以根据需要使用不同分子量 的壳寡糖， 在本发明的一个实施方案中， 所述壳寡糖的分子量小 于 5000。 在本发明进一步的实施方案中， 所述壳寡糖的分子量小 于 3000。 在本发明再进一步的实施方案中， 所述壳寡糖的分子量 小于 2000。 在本发明更进一步的实施方案中， 所述壳寡糖的分子 量小于 1500。 在本发明滴眼液的一个实施方案中， 其中含有 0.01~l%(w/v)的壳寡糖。 在本发明滴眼液的另一个实施方案中， 其中含有 0.01~0.5%(w/v)的壳寡糖。 在本发明滴眼液的又一个实 施方案中， 其中含有 0.05~0.2%(w/v)的壳寡糖。 在本发明滴眼液 的再一个实施方案中， 其中含有 0.05或者 0.1%(w/v)的壳寡糖。
本发明的滴眼液为一种水性组合物， 其中使用水作为该组合 物的介盾或溶媒或赋形剂或载体。 尽管本发明的滴眼液没有具体 指出所用的水占滴眼液总重量的百分数， 或者在配制本发明滴眼 液时没有具体指出所用水的量， 但是， 本领域技术人员清楚， 作 为滴眼液的介盾或溶媒或赋形剂或载体， 该水的量是以水加至滴 眼液全量来计算的， 这在液体制剂例如滴眼液、 注射液配制过程 中一般采用的计量方式。
在本发明的滴眼液中， 所述的抑菌剂可以是一种或多种的任 意组合。 在一个实施方案中， 所述的抑菌剂是苯氧乙醇和对羟基 苯曱酸乙酯的组合； 在进一步的实施方案中， 所述的抑菌剂是苯 氧乙醇和对羟基苯曱酸乙酯以任意比例的组合； 在再进一步的实 施方案中， 所述的抑菌剂是苯氧乙醇和对羟基苯曱酸乙酯以重量 比 50: 1的比例组合。 在一个实施方案中， 所述的抑菌剂是苯氧 乙醇和苯扎氯铵的组合； 在进一步的实施方案中， 所述的抑菌剂 是苯氧乙醇和苯扎氯铵以任意比例的组合； 在再进一步的实施方 案中， 所述的抑菌剂是苯氧乙醇和苯扎氯铵以重量比 50: 1的比 例组合。 在一个实施方案中， 所述的抑菌剂是三氯叔丁醇和苯扎 氯铵的组合； 在进一步的实施方案中， 所述的抑菌剂是三氯叔丁 醇和苯扎氯铵以任意比例的组合； 在再进一步的实施方案中， 所 述的抑菌剂是三氯叔丁醇和苯扎氯铵以重量比 10: 1的比例组合。 在一个实施方案中， 所述的抑菌剂是三氯叔丁醇和对羟基苯甲酸 乙酯的组合； 在进一步的实施方案中， 所述的抑菌剂是三氯叔丁 醇和对羟基苯曱酸乙酯以任意比例的组合; 在再进一步的实施方 案中， 所述的抑菌剂是三氯叔丁醇和对羟基苯曱酸乙酯以重量比 10: 1的比例组合。
本发明第二方面还可以提供这样的滴眼液， 即， 其包含本发 明第一方面任一项所述的滴眼液， 其中还可以含有药学可接受的 赋形剂， 例如可以将本发明第一方面的滴眼液在临床使用前用适 当的溶媒稀释成本发明第二方面的即用型滴眼液， 以获得某种临 床适用的效果， 例如本发明第一方面的滴眼液长期保存所适用环 境 pH与生理 pH不一致时， 可以在临用前向本发明第一方面的 滴眼液中加入适量的 pH调节剂， 以便使获得的本发明第二方面 的滴眼液的 pH值调节到眼部生理可接受的 pH环境下。
本发明进一步涉及具有本发明实施例配方的滴眼液。
本发明进一步涉及具有以下配方的滴眼液：
小牛血去蛋白提取物 200重量份， 苯氧乙醇 5重量份， 苯扎氯铵 0.1重量份， 壳寡糖 0.5重量份，
Figure imgf000008_0001
本发明进一步涉及具有以下配方的滴眼液:
小牛血去蛋白提取物 200重量份， 苯氧乙醇 5重量份， 羟苯乙酯 0.1重量份， 壳寡糖 0.5重量份，
Figure imgf000008_0002
本发明进一步涉及具有以下配方的滴眼液:
小牛血去蛋白提取物 200重量份， 三氯叔丁醇 1重量份， 苯扎氯铵 0.1重量份， 壳寡糖 1.0重量份，
Figure imgf000008_0003
本发明进一步涉及具有以下配方的滴眼液：
小牛血去蛋白提取物 200重量份， 三氯叔丁醇 1.0重量份， 羟苯乙酯 0.1重量份， 壳寡糖 1.0重量份，
注射用 适量， 至 1000重量份。
本发明第三方面还提供了本发明第一方面任一项所述滴眼液 或本发明第二方面任一项所述滴眼液在制备用于治疗眼病 (例如， 角膜病例如角膜损伤， 结膜病变， 干眼症)的医药中的用途。
本发明第四方面提供了一种治疗和 /或预防眼病 (例如，角膜病 例如角膜损伤， 结膜病变， 干眼症)的方法， 其包括给有此需要的 受试者施用治疗和 /或预防有效量的本发明第一方面任一项所述滴 眼液或本发明第二方面任一项所述滴眼液。
如本文使用的， 术语"受试者 "是指包括但不限于哺乳动物例 如猪、 狗、 猴、 猫、 人等以及禽类例如鸡、 鸭等， 优选地， 所述 受试者是人， 特别是罹患或可能罹患本文所述眼病 (例如， 角膜病 例如角膜损伤， 结膜病变， 干眼症)的人。
本发明的滴眼液可采用如下工艺制备： 在无菌条件下称取处 方量壳寡糖 (以及， 必要时， 还可包括抑菌剂例如苯氧乙醇和羟苯 乙酯的组合、 pH调节剂、 渗透压调节剂、 粘度调节剂等)， 用适 量注射用水溶解， 加入小牛血去蛋白提取物， 搅拌均勾， 加入注 射用水至近全量。 先用 0.45μιη微孔滤膜过滤， 再补加注射用水 至全量， 最后用 0.22μιη微孔滤膜过滤除菌即得。 根据本发明制 得的小牛血去蛋白提取物的滴眼液， 小牛血来源固定、质量稳定， 生产工艺易于控制。 制得的小牛血去蛋白提取物眼用液体制剂外 观澄明、 无菌、 刺激性低、 毒性低， 可以为临床提供一种安全、 有效的眼用制剂。
本发明提供的小牛血去蛋白提取物的滴眼液能通过增加线粒 体的呼吸能力和高能磷酸的合成而促进细胞对氧的利用， 并能维 持人体内重要酶的生理功能， 因此， 它具有改善组织营养、 刺激 细胞再生和加速组织修复的作用， 并能使过度增生的肉芽组织蜕 变， 胶原纤维重组， 减少或避免瘢痕形成。 小牛血去蛋白提取物 的滴眼液表面活性好， 眼部耐受性佳， 它可在角结膜表面均匀分 布， 从而润滑角膜， 减少眼睑对角膜表面的机械摩擦， 减轻局部 刺激症状。故对于角膜病尤其是角膜损伤， 在常规治疗的基础上， 联合应用小牛血去蛋白提取物的滴眼液， 可促进角膜上皮的生长 修复， 加速损伤愈合。
本发明提供的小牛血去蛋白提取物的滴眼液，可以用于治疗角 膜和结膜病变， 亦可用于干眼症， 给创伤性、 炎症性和营养性角 膜、 结膜病变及干眼症等的治疗带来新的选择。 具体实施方式：
下面通过具体的实施例进一步说明本发明， 但是， 应当理解 为， 这些实施例仅仅是用于更详细具体地说明之用， 而不应理解 为用于以任何形式限制本发明。
本发明对试验中所使用到的材料以及试验方法进行一般性和 /或具体的描述。 虽然为实现本发明目的所使用的许多材料和操作 方法是本领域公知的， 但是本发明仍然在此作尽可能详细描述。 本领域技术人员清楚， 在下文中， 如果未特别说明， 本发明所用 材料和操作方法是本领域公知的。
如未特别说明， 本发明以下实施例中所用的小牛血去蛋白提 取物中总固含量均不少于 35mg/ml。 实施例 1
处方：
小牛血去蛋白提取物 200ml 苯氧乙醇 5.0g
苯扎氯铵 O.lg
壳寡糖 0.5g
注射用水 适量
1000ml
制备工艺：
在无菌条件下称取处方量的苯氧乙醇、 苯扎氯铵、 壳寡糖用 适量注射用水加热溶解， 冷却至室温加入小牛血去蛋白提取物与 注射用水至全量。 先用 0.45μιη微孔滤膜过滤， 之后用 0.22μιη微 孔滤膜过滤除菌， 灌装， 即得滴眼液。 本实施例中所用壳寡糖的 分子量为 5000以下。 实施例 2
处方：
小牛血去蛋白提取物 200ml
苯氧乙醇 5.0g
羟苯乙酯 O.lg
壳寡糖 0.5g
Figure imgf000011_0001
制备工艺：
在无菌条件下称取处方量的苯氧乙醇、 羟苯乙酯、 壳寡糖用 适量注射用水加热溶解， 冷却至室温加入小牛血去蛋白提取物与 注射用水至全量。 先用 0.45μιη微孔滤膜过滤， 之后用 0.22μιη微 孔滤膜过滤除菌， 灌装， 即得滴眼液。 本实施例中所用壳寡糖的 分子量为 3000以下。 实施例 3
处方:
小牛血去蛋白提取物 200ml
三氯叔丁醇
苯扎氯铵 o.ig
壳寡糖
注射用 7J
1000ml
制备工艺：
在无菌条件下称取处方量的三氯叔丁醇、苯扎氯铵、 壳寡糖， 用适量注射用水加热溶解， 冷却至室温加入小牛血去蛋白提取物 与注射用水至全量。 先用 0.45μιη微孔滤膜过滤， 之后用 0.22μιη 微孔滤膜过滤除菌， 濯装， 即得滴眼液。 本实施例中所用壳寡糖 的分子量为 1500以下。 实施例 4
处方：
小牛血去蛋白提取物 200ml
三氯叔丁醇 l.Og
羟苯乙酯 O.lg
壳寡糖 l.Og
Figure imgf000012_0001
制备工艺：
在无菌条件下称取处方量的三氯叔丁醇、羟苯乙酯、 壳寡糖， 用适量注射用水加热溶解， 冷却至室温加入小牛血去蛋白提取物 与注射用水至全量。 先用 0.45μιη微孔滤膜过滤， 之后用 0.22μιη 微孔滤膜过滤除菌， 灌装， 即得滴眼液。 本实施例中所用壳寡糖 的分子量为 1500以下。 对照实施例 1
处方：
小牛血去蛋白提取物 200ml
三氯叔丁醇 l.Og
苯扎氯铵 O.lg
注射用水 t±
全量 1000ml 制备工艺： 除了不加壳寡糖外， 其它操作同实施例 3， 灌装 成滴眼液0 对照实施例 2
处方：
小牛血去蛋白提取物 200ml
三氯叔丁醇 l.Og
羟苯乙酯 O.lg
注射用 ti
全量 1000ml
制备工艺： 除了不加壳寡糖外， 其它操作同实施例 4, 濯装 成滴眼液。 眼刺激性实验：
ί一)、 实验材料
受试药物： 本发明实施例 1的滴眼液， 滴眼液的空白对照液， 本公司提供。
动物： 家兔， 曰本大耳白种， 体重 2.0 ~ 3.0kg， 雌雄兼用， 由锦州医学院实验动物中心提供。
(二)、 实验方法
多次给药对兔眼刺激性的观察： 取健康家兔 6只， 试验前 24 小时内对每只动物的双眼进行检查， 经检查选无眼刺激症状、 角 膜缺陷、 角膜损伤的家兔进行试验。 雌雄兼用， 每只眼睛于眼结 膜嚢内一侧给以小牛血去蛋白提取物眼用液体制剂， 另一侧给以 空白液作为对照， 滴入量为 O.lml, 每日给药 4 次， 连续给药 Ί 日， 使眼睑被动闭合约 10秒。 每日给药前及末次给药后 6、 24、 48、 72小时观察眼的局部反应情况。 与对照眼进行比较， 结果见 表 1。
表 1 多次给药眼刺激试验检查结果 (X±SD)(n = 4)
Figure imgf000014_0001
从表中结果可见， 给药组及对照组给药期及给药后 6、 24、 48、 72小时无异常， 评分为 0, 按标准判断本品无刺激性。
(三）、 结论： 小牛血去蛋白提取物的滴眼液多次给药家兔无 眼刺激作用 局部皮肤过敏实验
(-) , 实验材料
受试药物： 本发明实施例 1的滴眼液， 滴眼液的空白对照液， 阳性致敏物 (2，4-二硝基氯代苯)由本公司提供。
动物： 白色豚鼠， 体重 250 ~ 300g， 雌雄兼用， 由锦州医学 院实验动物中心提供。
(二)、 实验方法
取白色豚鼠 30只， 随机分为 3组， 每组 10只， 雌雄兼用。 给药前 24h将豚鼠背部两侧脱毛。 其中 I组给予本发明滴眼液； II组给予空白对照液， m组给予阳性致敏物。
致敏接触： 将眼用液体制剂 0.2ml涂在豚鼠左侧脱毛区， 持 续 6h。 第 7d和第 14d， 以同样方法重复一次。 空白对照液、 阳 性致敏物组方法同眼用液体制剂组。
激发接触： 于末次给予眼用液体制剂致敏后 14d， 将眼用液 体制剂 0.2ml涂于豚鼠背部右侧脱毛区， 6h后去掉眼用液体制剂， 即刻观察， 然后于 24、 48、 72h再次观察皮肤过敏反应情况， 按 《新药临床前安全性评价与实践》 中制剂常规安全试验的评分标 准进行评分， 皮肤过敏反应评分标准见表 2， 致敏率分类见表 3。 空白对照液、 阳性致敏物组方法同眼用液体制剂组。 皮肤过敏反应评分标准
Figure imgf000016_0001
红斑形成总分 +水肿形成总分 反 平均值 =
合计的动物数 致敏率分类
Figure imgf000016_0002
结果： 皮肤过敏反应评分结果见表 4。
'J、牛血去蛋白提取物滴眼液局部皮肤过敏试验结果
Figure imgf000017_0001
将小牛血去蛋白提取物滴眼液涂于豚鼠皮肤进行致敏接触和 激发接触， 无红斑和水肿出现， 与空白对照液组无差别， 阳性致 敏物组致敏率为 100% , 表明本品无皮肤局部致敏性。
ί三)、 结论：
小牛血去蛋白提取物眼用液体制剂给药豚鼠无局部致敏性。 稳定性实施例：
ί一)、 实验材料
受试药物： 本发明实施例 3和 4及对照实施例 1和 2(分别简 称对照 1和对照 2)的滴眼液， 本公司提供。
(二)、 考察项目
pH值测定：参照中国药典 2005年版二部附录 VI Η操作。 pH 在 6.0-9.0之间。
可见异物： 参照中国药典 2005年版二部附录 C Η可见异物 检查法进行检查。
活力测定： 取本品， 采用瓦氏呼吸检压仪测定豚鼠肝匀浆的 呼吸活力 Q02， 呼吸活力应不小于 4.0 l O2/mg'h。
多肽含量： 参照国家药品标准第十六册 16-119 福林酚测定 法， 分光光度法测定。
ί三）、 实验方法
取受试滴眼液， 于温度 25。C±2。C ,相对湿度 60%±5%的条件 下放置， 按照分别于 0个月、 3个月、 6个月、 9个月、 12个月、 16个月、 18个月、 20个月、 22个月、 24个月按考察项目进行检 测， 与 0月样品比较。 结果见表 5、 表 6。
表 5 稳定性考察实验结果（1 ) 项目 时间 0月 3月 6月 9月 12月 实施例 3 6.80 6.82 6.87 6.88 6.86 实施例 4 6.82 6.85 6.89 6.91 6.92 对照 1 6.84 6.86 6.88 6.85 6.87 pH值 对照 2 6.80 6.82 6.89 6.88 6.94 实施例 3 澄明 澄明 澄明 澄明 澄明 实施例 4 澄明 澄明 澄明 澄明 澄明 对照 1 澄明 澄明 澄明 澄明 澄明 可见异物 对照 2 澄明 澄明 澄明 澄明 澄明 实施例 3Q02 6.98 6.88 6.62 6.46 6.12 实施例 4Q02 6.95 6.87 6.60 6.44 6.15 活力测定 对照 1 Q02 6.97 6.76 6.37 5.69 4.84 Q。2 (μΐ/mg-h) 对照 2 Q02 6.98 6.79 6.31 5.61 4.80 实施例 3 0.237 0.237 0.239 0.238 0.236 实施例 4 0.24 0.241 0.238 0.238 0.237 多肽含量 对照 1 0.238 0.237 0.238 0.236 0.236 对照 2 0.236 0.238 0.237 0.236 0.236 表 6 稳定性考察实验结果（2 )
Figure imgf000019_0001
依据实验要求， 活力测定要满足 Q02≥4。 从结果可见， 与不 使用壳寡糖相比， 滴眼液在 18月后蛋白活力仍满足要求。说明壳 寡糖可在一定程度上保护蛋白呼吸活力， 使得制剂更加稳定。 因 此， 通过稳定性实验结果证明， 加入壳寡糖， 使得本发明滴眼液 的稳定性明显提高。

Claims

权 利 要 求
1、 一种滴眼液， 其包含治疗有效量的小牛血去蛋白提取物、 壳寡糖、 和水。
2、 权利要求 1所述的滴眼液， 按该滴眼液的总量计， 其包含 10-30% (v/v)的小牛血去蛋白提取物、 0.01~1% (w/v)的壳寡糖、 和水。
3、 权利要求 2所述的滴眼液， 其中所述壳寡糖的分子量小于 5000， 优选小于 3000， 优选小于 2000， 优选小于 1500。
4、 权利要求 1至 3任一项所述的滴眼液， 其还包含一种或多 种选自以下的抑菌剂： 苯乙醇、 苯氧乙醇、 对羟基苯曱酸甲酯、 对羟基苯甲酸乙酯、 对羟基苯曱酸丙酯、 苯扎氯铵、 三氯叔丁醇。
5、 权利要求 1至 4任一项所述的滴眼液， 其还包含 pH调节 剂。
6、 权利要求 1至 5任一项所述的滴眼液， 其还包含渗透压调 节剂。
7、 权利要求 1至 6任一项所述的滴眼液， 其还包含粘度调节 剂。
8、 权利要求 1至 7任一项所述的滴眼液， 其具有以下任一项 的配方:
i)
小牛血去蛋白提取物 200重量份， 苯氧乙醇 5重量份， 苯扎氯铵 0.1重量份， 壳寡糖 0.5重量份，
Figure imgf000021_0001
小牛血去蛋白提取物 200重量份， 苯氧乙醇 5重量份， 羟苯乙酯 0.1重量份， 壳寡糖 0.5重量份，
注射用水 适量， 至 1000重量份; 小牛血去蛋白提取物 200重量份， 三氯叔丁醇 1重量份， 苯扎氯铵 0.1重量份， 壳寡糖 1.0重量份，
注射用水 适量， 至 1000重量份； 或者 小牛血去蛋白提取物 200重量份, 三氯叔丁醇 1.0重量份， 羟苯乙酯 0.1重量份， 壳寡糖 1.0重量份，
注射用水 至 1000重量
9、 权利要求 1至 8任一项所述滴眼液在制备用于治疗眼病 (例 如， 角膜病例如角膜损伤， 结膜病变， 干眼症)的医药中的用途。
10、 治疗和 /或预防眼病 (例如， 角膜病例如角膜损伤， 结膜病 变， 干眼症)的方法， 其包括给有此需要的受试者施用治疗和 /或预 防有效量的权利要求 1至 8任一项所述滴眼液。
PCT/CN2009/001475 2009-01-20 2009-12-16 小牛血去蛋白提取物的滴眼液 WO2010083636A1 (zh)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US13/145,311 US20120003323A1 (en) 2009-01-20 2009-12-16 Eye drops containing a deproteinized calf blood extract
EP09838603.0A EP2389924B1 (en) 2009-01-20 2009-12-16 An eyedrops of the deproteinized calf blood extract

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN2009100008939A CN101780105B (zh) 2009-01-20 2009-01-20 小牛血去蛋白提取物的滴眼液
CN200910000893.9 2009-01-20

Publications (1)

Publication Number Publication Date
WO2010083636A1 true WO2010083636A1 (zh) 2010-07-29

Family

ID=42355481

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2009/001475 WO2010083636A1 (zh) 2009-01-20 2009-12-16 小牛血去蛋白提取物的滴眼液

Country Status (4)

Country Link
US (1) US20120003323A1 (zh)
EP (1) EP2389924B1 (zh)
CN (1) CN101780105B (zh)
WO (1) WO2010083636A1 (zh)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102579492A (zh) * 2012-03-05 2012-07-18 兆科药业（广州）有限公司 一种含小牛血去蛋白提取物的玻璃酸钠滴眼液及其制备方法
CN104147047B (zh) * 2014-07-30 2017-10-20 哈尔滨圣泰生物制药有限公司 一种小牛血去蛋白提取物组合物及其注射剂和制法
CN111671719A (zh) * 2020-06-18 2020-09-18 广东药科大学 一种壳寡糖组合物及其制备方法和应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1486701A (zh) * 2002-10-01 2004-04-07 青岛海洋大学 一种人工泪液
CN1586499A (zh) * 2004-07-08 2005-03-02 合肥兆峰科大药业有限公司 小牛血去蛋白提取物、其眼膏制剂及其制备方法
CN1895275A (zh) * 2005-07-12 2007-01-17 锦州奥鸿药业有限责任公司 一种小牛血去蛋白提取物凝胶剂

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS60149521A (ja) * 1984-01-18 1985-08-07 Eisai Co Ltd 高眼圧・緑内障治療・予防用点眼剤
US5422116A (en) * 1994-02-18 1995-06-06 Ciba-Geigy Corporation Liquid ophthalmic sustained release delivery system
FR2823441B1 (fr) * 2001-04-12 2004-09-10 Thea Lab Composition pharmaceutique a base de macrolides pour application locale en ophtalmologie et son procede de preparation
FR2841137B1 (fr) * 2002-06-20 2004-08-13 Bioalliance Pharma Systeme de vectorisation comprenant des nanoparticules de taille homogene d'au moins un polymere et d'au moins un polysaccharide charge positivement
WO2006094027A2 (en) * 2005-03-02 2006-09-08 Nascent Pharmaceutical, Inc. Combination therapy for topical application
EP2425816A3 (en) * 2007-01-31 2014-04-30 Allergan, Inc. A pharmaceutical composition for use in treating an ocular disease or condition and a method for producing the composition

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1486701A (zh) * 2002-10-01 2004-04-07 青岛海洋大学 一种人工泪液
CN1586499A (zh) * 2004-07-08 2005-03-02 合肥兆峰科大药业有限公司 小牛血去蛋白提取物、其眼膏制剂及其制备方法
CN1895275A (zh) * 2005-07-12 2007-01-17 锦州奥鸿药业有限责任公司 一种小牛血去蛋白提取物凝胶剂

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP2389924A4 *

Also Published As

Publication number Publication date
CN101780105A (zh) 2010-07-21
US20120003323A1 (en) 2012-01-05
EP2389924A4 (en) 2013-12-18
CN101780105B (zh) 2011-09-28
EP2389924B1 (en) 2014-08-27
EP2389924A1 (en) 2011-11-30

Similar Documents

Publication Publication Date Title
JP2611159B2 (ja) ヒアルロン酸薬理活性画分、その製造方法および医薬組成物
JPH09169667A (ja) 新規医薬組成物
Chebbi et al. Pilot study of a new matrix therapy agent (RGTA OTR4120) in treatment-resistant corneal ulcers and corneal dystrophy
WO2007036107A1 (fr) Formulation à application externe et ses procédés de préparation et utilisations
US11534459B2 (en) Compositions and methods of treating dry eye syndrome and other traumatized non-keratinized epithelial surfaces
Wang et al. Lollipop‐inspired multilayered drug delivery hydrogel for dual effective, long‐term, and NIR‐defined glaucoma treatment
CN114425033B (zh) 一种含有间充质干细胞外泌体的眼用凝胶及其制备方法
WO2010083636A1 (zh) 小牛血去蛋白提取物的滴眼液
US9314446B2 (en) Sustained releasing pharmaceutical composition
CN102188695B (zh) 一种眼用凝胶组合物
RU2477637C2 (ru) Способ лечения острого нарушения мозгового кровообращения ишемического и геморрагического характера
WO2023272813A1 (zh) 一种温度敏感型凝胶损伤修复制剂及其应用
CN112891326B (zh) 一种载有那他霉素的海藻酸凝胶药膜及其制备方法
KR100963611B1 (ko) 알긴산류 화합물을 포함하는 점안용 조성물 및 그의제조방법
CN113786380A (zh) 一种硝酸毛果芸香碱眼用凝胶及其制备方法
WO2021078246A1 (zh) 一种用于预防或治疗脓毒症的药物组合物、试剂盒及其应用和治疗方法
CN112957374A (zh) 一种犬用犬血清去蛋白眼睛凝胶及其制备方法
JP2008511652A (ja) 創傷治癒の促進方法
Regu et al. Ocular Delivery of Metformin for Sustained Release and in Vivo Efficacy
WO2024067874A1 (zh) 一种改性壳聚糖及其在大分子活性成分递送中的应用
CN115212200B (zh) 用于治疗糖尿病并发症的含葛根素的复方制剂及其制备方法
CN116726151B (zh) 一种用于角膜损伤修复的日抛型滴眼液及其制备方法
RU2362554C2 (ru) Регенерирующее противовоспалительное средство и способы лечения с помощью этого средства
RU2733392C1 (ru) Комбинированное офтальмологическое средство
CN113143850A (zh) 一种猫用眼睛凝胶及其制备方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09838603

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2009838603

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 13145311

Country of ref document: US