KR20130006110A - Pharmaceutical composition for treatment of recurrent corneal erosion comprising umbilical cord serum - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing umbilical cord serum and a method for manufacturing the same are provided to effectively treat recurrent corneal erosion without side effects, and to develop a therapeutic agent for treating ophthalmologic diseases. CONSTITUTION: A pharmaceutical composition for preventing or treating recurrent corneal erosion contains 18-22%(v/v) of umbilical cord serum. The composition is a liquid or gel type for instillation. The pharmaceutical composition additionally contains a pharmaceutically acceptable carrier. A method for manufacturing the composition comprises: a step of collecting umbilical cord blood; a step of collecting serum from the umbilical cord blood; and a step of diluting the umbilical cord serum using a balanced salt solution. The umbilical cord blood is collected from umbilical vein of a mother who gave birth.

Description

Pharmaceutical composition for treatment of recurrent corneal erosion comprising umbilical cord serum}

The present invention relates to a pharmaceutical composition for the prevention or treatment of recurrent corneal erosion, comprising a umbilical cord serum, and a method for preparing the same.

Recurrent corneal erosion syndrome is defined as the repeated induction of corneal epithelial ablation accompanied by pain, redness, tearing and potential blindness. Recurrent corneal erosion syndrome is either endogenous from trauma to the corneal epithelium or endogenously in the context of epithelial dystrophy such as epithelial basement membrane dystrophy, Bowman layer dystrophy such as Reis-Buckler dystrophy, and interstitial dystrophy such as granular dystrophy or lattice dystrophy. May occur. It is associated with changes in the corneal epithelial basement membrane, Bowman layer and corneal stroma, based on ultrastructural pathology characterized by abnormal epithelial basement membrane, absence or abnormality of semi-colloid, and loss of fixed fibril. Treatment of recurrent corneal erosion is known to be very difficult and involves surgery including lubricants, high osmotic agents, eye contact lenses and corneal interstitial punctures, mechanical corneal epithelial debridement, therapeutic laser keratectomy and alcohol (plate / ectoderm) separation. Attempts have been made to manage recurrent corneal erosion through medical care. However, the development of a therapeutic agent that suppresses the recurrence of recurrent corneal erosion and can completely cure it has been insufficient.

Autologous eye drops are currently commercially available eye drops with serum separated from the patient's own blood, so the patient's own serum does not cause allergic or rejection reactions and has the advantage of containing antibodies that protect antibodies and bacteria. In addition, serum contains epithelial growth factor (EGF), vitamin A, transforming growth factor (TGF) -β, acidic and basic fibroblast growth factor, platelet-derived growth factor, fibrinogen, and Neurotrophs such as many growth factors and tear components including serum antiproteinases such as α2 macroglobulin, substance P, insulin-like growth factor-1 (IGF-1) and nerve growth factor Contains substance Serum oil and whole albumin can also lubricate and nourish the ocular surface epithelium. However, long-term use of autologous serum may cause discomfort and rejection of treatment due to the hassle of continuously repeated blood sampling from patients, and it is not possible to use it for patients with anemia, which is difficult to collect. Therefore, it is necessary to develop an eye drop that can be easily produced and has excellent therapeutic effect of recurrent corneal erosion.

Accordingly, the present inventors have made intensive studies to develop a safer pharmaceutical composition for treating ophthalmic diseases, and as a result, when using an ophthalmic pharmaceutical composition containing cord serum, it is possible to effectively treat recurrent corneal erosion without side effects. The present invention has been completed.

It is a main object of the present invention to provide a pharmaceutical composition for the prevention or treatment of recurrent corneal erosion comprising umbilical cord serum.

Another object of the present invention is to provide a method for preparing the pharmaceutical composition.

As one embodiment for achieving the object of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of recurrent corneal erosion, including umbilical cord serum.

In another aspect, the present invention provides a method for preventing or treating recurrent corneal erosion, comprising administering to the individual a therapeutically effective amount of an eye drop composition comprising a cord serum.

The term "umbilical cord serum" of the present invention refers to serum separated from the umbilical cord of the fetus, preferably collected from the umbilical cord discarded during birth or stillbirth of the fetus or from the mother's umbilical vein after birth can do.

The pharmaceutical composition of the present invention is preferably a pharmaceutical composition for eye drops.

The term "eye drop pharmaceutical composition" of the present invention means a pharmaceutical composition in the form of a drop medicine containing an active ingredient exhibiting a desired therapeutic effect based on physiological saline, which can be administered to the eye. The active ingredient is not particularly limited thereto, but may be a steroid, an antihistamine, a sympathetic stimulant, a parasympathetic stimulant, a prostaglandin, a nonsteroidal anti-inflammatory agent, a local anesthetic, or the like in the present invention. Include. At this time, the content of the umbilical cord serum contained in the eye drop pharmaceutical composition of the present invention is not particularly limited, but is preferably included in 18 to 22% (v / v).

Meanwhile, the ophthalmic pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, but the carrier is not particularly limited thereto, and may be a preservative, an isotonic agent, a stabilizer, a pH adjuster, and the like. Follows the range of general pH, osmotic pressure and viscosity of the ophthalmic composition. Examples of the preservatives include benzalkonium chloride 0.002 to 0.1% (w / v), cerimide 0.002 to 0.01% (w / v), chimerosal 0.001 to 0.01% (w / v), and chlorobutanol 0.25 to 0.5% (w / v ), Polyhexamethylene biguanide 0.002 to 0.1% (w / v), paraoxybenzoic acid alkyl methyl, paraoxybenzoic acid ethyl, paraoxybenzoic acid propyl or paraoxybenzoic acid butyl 0.001 to 0.05% (w / v), dihydro Acetic acid 0.001 to 0.1% (w / v), chlorocresol 0.0001 to 0.05% (w / v), chlorohexidine 0.001 to 0.01% (w / v) and the like can be used, the isotonic agent is not particularly limited However, D-sorbitol, glycerin, concentrated glycerin, mannitol and maltitol syrup, ionic substances, and the like, which can exhibit an osmotic pressure of 270 to 310 mOsmol / kg of physiological saline, may be used, and the content of these isotonic agents is particularly limited thereto. But preferably in a weight ratio to the volume of the pharmaceutical composition. 5 to 20%, more preferably 1 to 10%. In addition, the stabilizers include sodium etate (0.05 to 0.2% (w / v)) and ε-aminocaproic acid (0.01 to 0.05% (w / v)), anhydrous sodium sulfate (0.01 to 0.2% (w / v) ), Ascorbic acid (0.02 to 0.5% (w / v)), citric acid (0.3 to 2.0% (w / v)) and the like can be used, pH regulators are not particularly limited, pH of the pharmaceutical composition is 5.0 to 8.0 hydrochloric acid, sodium hydroxide, buffers and the like can be used.

The type of formulation of the pharmaceutical composition of the present invention is not limited, but is preferably in the form of eye drops or gels.

As used herein, the term "recurrent corneal erosion" refers to an ocular disease characterized by damage to the outermost layer of the cornea of epithelial cells attached to the basement membrane (the Bowman layer) as a disease commonly seen in the clinic. The corneal epithelium is peeled off in a rash and is characterized by a recurrent disease in which the process of re-epithelialization occurs after the re-epithelialization at the peeled site. The disease is caused by a previous history of corneal injury such as corneal abrasion, corneal dystrophy, or corneal disease, and because of the damage of the epithelial cells, sensitive corneal nerves are exposed, resulting in severe pain, in addition to foreign bodies, Symptoms include glare and recurrent onset of tears.

As used herein, the term "prevention" means any action that inhibits or delays the symptoms of recurrent corneal erosion by administration of the composition. In the present invention, in an individual who has already experienced a recurring corneal erosion and has been treated, the act of inhibiting or delaying the recurring corneal erosion does not occur again.

In addition, the term "treatment" in the present invention means any action that improves or advantageously changes the symptoms of recurrent corneal erosion by administration of the composition. The present invention also includes acts to alleviate the symptoms of recurrent corneal erosion, and reduce the number of recurrences and the frequency of recurrences.

In order to treat or improve the recurrent corneal erosion, in general, a sufficiently viscous tear-shaped liquid is supplied, wherein the liquid contains glycoproteins rather than water or saline, and has high wettability. Preference is given to using. In the present invention, the cord serum for treating the recurrent corneal erosion is included in an ophthalmic pharmaceutical composition.

The recurrent corneal erosion causes discomfort and irritation due to the lack of tears and damage to the eye surface of the gap of the eyelids exposed by excessive evaporation of the tear film, causing dry eye or inflammation of the eye surface with inflammation of the eye surface, The mechanism of development is different from neurotrophic keratitis, a disease in which corneal epithelium is not healed due to deterioration, and treatment methods are also different.

According to a specific embodiment of the present invention, the present inventors have a cord serum containing a high concentration of essential tear factors and many growth factors such as EGF, vitamin A and TGF-b and neurotrophic factors such as substances P, IGF-1 and NGF Dilutions were used with artificial tears to enhance the therapeutic effect of recurrent corneal erosion.

More specifically, the efficacy of the ophthalmic pharmaceutical composition comprising umbilical cord serum prepared in the present invention was evaluated in patients with severe recurrent corneal erosion, which frequently experienced recurrence of corneal erosion. As a result, the number and frequency of recurrence of recurrent corneal erosion were remarkably reduced in the experimental group administered with the umbilical cord serum eye drops prepared in the present invention, compared with the control group administered with only artificial tears. In the group administered eye drops containing the cord serum prepared in the present invention, recurrence of recurrent corneal erosion occurred only in 6 of 18 patients, and its frequency was low, such as twice and once. On the other hand, recurrence of recurrent corneal erosion occurred in 16 of 17 patients in the group receiving artificial tears only, and their frequency was high 5 times and 3 times. That is, the umbilical cord serum eye drops of the present invention significantly reduce the frequency and frequency of recurrence of recurrent corneal erosion, and can also be used harmless to the human body (Example 2-3).

In addition, the ophthalmic solution of the present invention is the use of umbilical cord eye drops instead of the known autologous eye drops, which may cause discomfort and rejection of treatment due to the hassle of continuously repeated blood collection from the patient when the autologous serum is used for a long period of time. Can overcome. In other words, since cord serum can obtain a large amount of serum from the umbilical vein at one time and can be used to prepare cord serum eye drops in advance, it is useful for the treatment of chronic ocular surface disease that requires long-term use, It can be useful and can reduce the risk of fainting, bruising and infection due to blood collection.

The composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level refers to the type and severity of the subject, the severity, age, sex and activity of the drug. , Drug sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs, and other factors well known in the medical arts.

The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. And single or multiple administrations. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

In addition, according to another embodiment of the present invention, the present invention provides a method for preparing the pharmaceutical composition. Specifically, the method for producing a pharmaceutical composition of the present invention comprises the steps of: (i) collecting umbilical cord blood; (Ii) obtaining serum from said umbilical cord blood; And (iii) diluting the cord blood serum.

In this case, the cord blood serum is not particularly limited thereto, but may be obtained by collecting cord blood from mothers who are delivered by suffocation or cesarean section, and in one embodiment of the present invention, re-blood from the navel vein of the mother after fetal delivery is obtained. Collected. In addition, the present invention is not particularly limited thereto, and it is preferable to use the cord blood obtained by allowing the cord blood to stand at room temperature and coagulating the blood.

As a method of separating cord serum from collected cord blood, the time of coagulation at room temperature and the speed and time of centrifugation are important. If the time of coagulation at room temperature is short or the centrifugation time is short, low purity serum can be obtained. . Therefore, the blood coagulation time is preferably about 1 hour 30 minutes to 3 hours, the centrifugation time is about 10 minutes to 30 minutes, and the centrifugation speed is about 2,000 g to 4,000 g. In an embodiment of the present invention, the coagulation time of the cord blood was about 2 hours, and the coagulated blood was centrifuged at 3,000 g for 15 minutes, and serum was separated therefrom and used for the experiment.

In the step of diluting the cord blood serum of step (iii), the type of the solution for diluting the cord blood is not limited, but is preferably diluted using a balanced salt solution. In addition, the cord blood serum is preferably diluted to 18 to 22% (v / v) relative to the final composition.

Since the pharmaceutical composition including the cord serum of the present invention can effectively treat recurrent corneal erosion without side effects, it may be widely used in the development of a therapeutic agent for eye diseases as well as safer treatment of eye diseases.

1 is before (A) and after (B) treatment of cord serum eye drops in patients with recurrent corneal erosion due to trauma, and before (C) and after treatment of cord serum eye drops in patients with recurrent corneal erosion in the context of epithelial dystrophy ( The state of D) is shown by a slit lamp microscope test result.

Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These embodiments are only for illustrating the present invention, and the scope of the present invention is not limited by these embodiments.

Example  One: Umbilical cord serum  Preparation of ophthalmic pharmaceutical composition containing

Example  1-1: Cord serum  purchase

After obtaining informed consent, cord blood was collected from mothers who were delivered by asphyxia or cesarean section. Experimental data for hepatitis B and C viruses and human immunodeficiency virus were examined twice at 8 and 38 weeks of pregnancy. After fetal delivery, a volume of 200-250 ml of cord blood was collected from the umbilical vein. The collected cord blood was coagulated for 2 hours at room temperature, and centrifuged at 3,000 g for 15 minutes, and serum was carefully separated under sterile conditions in an air laminar flow hood. Alternatively, the collected cord blood was coagulated for about 30 minutes at room temperature, and serum was separated by centrifugation at 1,500 g for 5 minutes, but the coagulation time was increased and the centrifugation was performed at a high speed. Serum was isolated in this manner as it was possible to obtain serum with higher purity when coagulated for 2 hours at room temperature and centrifuged at 3,000 g for 15 minutes.

Example  1-2: Umbilical cord serum  Preparation of eye drop containing pharmaceutical composition

Balanced salt solution (BSS) was added to the cord serum obtained in Example 1-1 and diluted to 20%, and a portion of the diluted serum was placed in a sterile 5 ml bottle of UV protection. Opened bottles were stored in a 4 ° C. refrigerator and unopened ones were stored in a -20 ° C. freezer. Opened bottles were discarded after one week of use and frozen for up to three months.

To the cord serum obtained in Example 1-1 was added a preservative normal saline solution (unpreserved normal saline solution) to dilute the cord serum to prepare an eye drop, but the efficacy of the eye drops lower than the case of using the equilibrium saline solution in the present invention Eye drops diluted with equilibrium saline were used for the experiment.

Example  2: Umbilical cord serum  Evaluation of Efficacy of Ophthalmic Pharmaceutical Compositions Containing

Example  2-1: Patient Selection

Severe macrofoam corneal erosion patients who had already experienced several recurrences of corneal erosion and showed normal results in the Schirmer test (10 mm) were selected. Individuals with a history of herpetic keratitis, diabetes or other systemic diseases were excluded from this study.

The efficacy of the ophthalmic pharmaceutical composition of the present invention was evaluated in 35 patients (35 eyes) with recurrent corneal erosion. Of the 35 patients with recurrent corneal erosion, 17 were male and 18 were female. The mean age was 46.6 ± 13.4 years and the mean follow up duration was 14.7 ± 2.5 months. Group A consisted of 18 patients (18 eyes) who received 20% cord serum eye drops with artificial tears between 2007 and 2008. Eight males and ten females were included. The mean age was 46.7 ± 12.1 years and the mean weight was 14.3 ± 2.7 months. Predisposing factors for recurrent erosion include ocular trauma in seven patients and epithelial basement membrane dystrophy in two patients. Nine patients did not have any global risk factors.

Group B consisted of 17 patients (17 eyes) who used artificial tears only between 2005 and 2006. Nine males and eight female patients were included. The mean age was 46.5 ± 15.0 years and the mean follow-up period was 15.1 ± 2.1 months. While six patients had a history of ocular trauma and three had a history of epithelial basement dystrophy, eight had no prognostic risk factors. No significant differences in age, sex, duration of follow-up and global risk factors were observed between the two groups. Prior consent was obtained from each subject enrolled in this study.

Example  2-2: Umbilical cord serum  Administration of eye drops containing pharmaceutical composition

Umbilical cord serum eye drops in group A were administered 4-6 times per day with preservative free tears. In group B, only preservative free tears were administered 4-6 times a day.

Eye examinations, including visual measurements and slit lamp biomicroscopy, were performed before and after treatment (FIG. 1). The analysis of the corneal state by slit lamp microscopy showed that corneal lesions improved after the use of umbilical cord serum in patients with recurrent corneal erosion (A) due to trauma. In patients with recurrent corneal erosion at the periphery of the cornea due to epithelial dystrophy, corneal lesions were completely lost after umbilical cord serum eye drops.

In addition, patients were instructed to visit the clinic if new symptoms developed. Relapse is defined as the presence of clinical symptoms involving epithelial defects in the cornea. The Mann-Whitney U test was used to compare the recurrence frequency between the two test groups.

Example  2-3: Umbilical cord serum  Effects of Eye Drops Containing Pharmaceutical Compositions

Table 1 and 2 show the characteristics and results of patients with recurrent corneal erosion in the group treated with eye drops containing umbilical cord serum (Group A) and the group not treated (Group B).

Characteristics and Results of Recurrent Corneal Erosion Patients Treated with Umbilical Cord Serum and Artificial Tear. Patient number age
(three) gender Risk factors Follow up period
(month) Recurrence Recurrence frequency
(Count / month) One 40 female Epithelial basement membrane dystrophy 15 One 0.07 2 24 south - 12 2 0.17 3 38 female credit 19 0 0 4 52 south - 14 0 0 5 58 south - 10 0 0 6 40 female - 14 0 0 7 33 female credit 16 0 0 8 60 south - 11 2 0.18 9 33 female - 18 0 0 10 45 female credit 16 0 0 11 50 south credit 14 0 0 12 67 south Epithelial basement membrane dystrophy 12 One 0.08 13 58 female credit 18 One 0.06 14 66 south credit 17 0 0 15 50 female - 15 0 0 16 35 female - 10 0 0 17 42 south credit 12 2 0.17 18 49 female - 15 0 0

Characteristics and Treatment Outcomes of Recurrent Corneal Erosion Patients Treated with Artificial Tears Only. Patient number age
(three) gender Risk factors Follow up period
(month) Recurrence Recurrence frequency
(Count / month) One 54 south Epithelial basement membrane dystrophy 18 5 0.29 2 58 south - 14 3 0.21 3 34 south credit 13 3 0.23 4 25 female - 15 2 0.13 5 45 female Epithelial basement membrane dystrophy 17 2 0.12 6 54 female - 16 2 0.13 7 54 south - 18 2 0.11 8 66 south credit 11 3 0.27 9 41 female Epithelial basement membrane dystrophy 15 3 0.20 10 30 female credit 14 One 0.07 11 79 female - 14 3 0.21 12 56 female credit 15 2 0.13 13 38 south credit 17 2 0.12 14 31 south - 13 One 0.08 15 27 female credit 17 2 0.12 16 42 south - 12 0 0 17 57 south - 18 2 0.11

Recurrence in Group A occurred twice in three eyes (16.7%) and once in the other three eyes (16.7%) and not in the remaining 12 eyes (66.7%). Group B occurred five times in one eye (5.9%), three in five eyes (29.4%), two in eight eyes (47.1%), and one in two eyes (11.8%). It did not occur in the eye (5.9%). The average frequency of relapse was 0.50 ± 0.79 (0.04 ± 0.07 / month) in group A and 2.24 ± 1.09 (0.15 ± 0.08 / month) in group B (p <0.01).

That is, when the ophthalmic solution containing the cord serum prepared in the present invention was administered to a patient suffering from severe recurrent corneal erosion, recurrence of corneal erosion was markedly reduced, which was not a level of temporary symptom relief but a complete therapeutic effect. It was to show that there is. In addition, the recurrence rate of recurrent corneal erosion was reduced to about 1/5 when compared to a patient who received only artificial tears, and thus, the excellent recurring corneal erosion effect of the eye drop containing the cord serum prepared in the present invention was improved. I could see that.

In addition, when administering eye drops containing umbilical cord serum to the patient, no noticeable complications associated with their use were observed. The eye drops of the present invention are safe for the human body and have excellent recurring corneal erosion without any side effects. It was found to have a therapeutic effect.

Claims (9)

A pharmaceutical composition for the prevention or treatment of recurrent corneal erosion comprising umbilical cord serum.
The composition of claim 1, wherein the composition is in the form of eye drops or gels.
The method according to claim 1 or 2,
The pharmaceutical composition of the cord serum is 18 to 22% (v / v).
The method according to claim 1 or 2,
A pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
(Iii) collecting umbilical cord blood;
(Ii) obtaining serum from said umbilical cord blood; And
(Iii) diluting the umbilical cord blood serum, wherein the composition of claim 1 or 2 is prepared.
The method of claim 5,
The umbilical cord blood is collected from a maternal umbilical vein.
The method of claim 5,
The cord blood serum is obtained by leaving the cord blood at room temperature to coagulate blood and centrifuge it.
The method of claim 5,
The cord blood serum is diluted using a balanced salt solution.
The method of claim 5,
Said cord blood serum is diluted to 18-22% (v / v) relative to the final composition.

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