JPS63126831A - Immunoglobulin formulation for eye drops - Google Patents

Abstract

PURPOSE:To obtain the titled formulation containing an immunoglobulin as an active ingredient and applied to patients having bacterial or viral eye-disease. CONSTITUTION:An immunoglobulin is dissolved in sterilized and distilled water as an active ingredient in a content of 0.1-15% based on the formulation and as necessary at least one auxiliary of buffer, isotonic agent (e.g.; boric acid, borax, sodium salt, potassium salt, etc.), thickener (e.g. polyvalent alcohols, polysaccharides; capable of enhancing drug effect by preventing overflow of eye drops and prolonging residence time in eyes), preservatives (e.g. paraoxybenzoic acid esters, cationic soaps, alcohol derivatives, etc.), stabilizer (e.g.; sodium sulfite, etc., for oxidant, sodium thiosulfate, etc., for color protecting agent, citric acid, etc., for decomposition-preventing agent), surfactant (e.g.; preferably nonionic surfactant such as fatty acid polyvalentalcoholpolyoxyethylene ether, etc.) is added thereto and pH is adjusted to 4-9 with hydrochloric acid, etc., to provide the aimed immunoglobulin formulation for eye drops.

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to an ophthalmic immunoglobulin preparation applied to affected areas of bacterial or viral ophthalmic diseases.

<Conventional technology> It is used as a therapy for internal ocular diseases such as uveal prolongation caused by bacteria or viruses, and external ocular diseases such as conjunctivitis and keratitis. Use eye drops. These eye drops often contain antibiotic substances.

<Problems to be Solved by the Invention> However, for the above-mentioned various diseases, sufficient therapeutic effects may not be obtained only with eye drops containing antibiotics. That is, antibiotics are of course ineffective against viral keratoconjunctivitis, acute hemorrhagic conjunctivitis, and corneal herpes caused by viruses, and are almost ineffective when it comes to severe eye infections, even those caused by bacterial inflammation.

Means for Solving the Problems> Therefore, in the process of research and development, the present inventors have made efforts to obtain sufficient therapeutic effects for the above-mentioned ophthalmological diseases that are not effective with antibiotics. We obtained the following knowledge.

■The eye's immune system is highly biased depending on its location.

That is, among the external parts of the eye, the eyelids and conjunctiva are rich in immunoglobulin and immune cells, and the lymphatic system and vascular system are well-developed, so these areas have a high ability to prevent infection. However, among the external parts of the eye, the cornea has a relatively high immunoglobulin content, but has no immune cells and has a low infection prevention function.

Furthermore, the content of immunoglobulin in the inner eye is very small (for example, the aqueous humor is 1/200 of that in serum).
, complement 1/1000), and there are no immune cells.

Therefore, the inner eye and cornea have a low ability to prevent infection.

■ Burial pancreatitis and severe eye infections such as keratoconjunctivitis,
The immunoglobulin content (neutralizing antibody titer) in the tear fluid of patients suffering from acute bloody pancreatitis, corneal herpes, etc. is significantly lower than that of normal individuals. γ-globulin in tear fluid of normal people is usually 500-1000 ng/mjl, whereas
That of the lachrymal fluid of affected patients is less than 200 ng/ml.

Based on the above findings (1) and (2), the present inventors have conceived of an eye drop immunoglobulin preparation that contains immunoglobulin as an active ingredient and is applicable to bacterial and viral ophthalmological diseases.

Although it is known that immunoglobulin is used for the prevention and treatment of infectious diseases, most of the uses are injections, and there are only a few examples of external use, as described in Japanese Patent Application Laid-open No. 110819/1983. It has only been introduced as a treatment for skin diseases.

<Embodiments> Hereinafter, embodiments of the present invention will be described in detail. In this specification, percentages indicating formulations are by weight unless otherwise specified.

The ophthalmic immunoglobulin preparation of the present invention is prepared by adding sterile distilled water to
Dissolve immunoglobulin as an active ingredient, add at least one of adjuvants such as tonicity agents, thickeners, preservatives, stabilizers, surfactants, etc. as necessary, and adjust the pH to 4 to 4 with hydrochloric acid etc. It was adjusted to 9.

In addition to normal large immunoglobulin, the immunoglobulin used in the present invention includes human immunoglobulin treated with pepsin, plasmin, sulfonation, polyethylene glycol, acid, and alkylation. The content of this immunoglobulin in the preparation is usually 0.
1 to 15% (preferably 0.5 to 5%).

In addition, buffering agents and isotonic agents are inorganic acids and inorganic salts that are added to create a buffer solution and to make the i-load solution isotonic with physiological saline, respectively. Also serves as part or all of the agent. Specifically, boric acid, borax, (anhydrous) phosphoric acid salt, (anhydrous) phosphoric acid di-salt, hydrogen carbonate, carbonate, calcium salt, sodium salt, potassium salt (the latter two are mainly (used for diaphragm), etc.

Thickeners are added to prevent the ultra-outflow of eye drops and extend their retention time in the eye to enhance their efficacy, and include polyhydric alcohols (ex, glycerin, macrogol), Examples include saccharides (ex, sorbitol), polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, and the like.

Preservatives are added for antiseptic and sterilization purposes, and include baloxin benzoate esters and anti-inflammatory soaps (ex.
, benzalkonium chloride) alcohol derivatives (ex,
chlorobutanol), organic acids and their salts, phenols, organic mercury agents (ex.

Thimerosal), etc.

Stabilizers are added to prevent oxidation, coloration, and decomposition when the main ingredient (immunoglobulin) is unstable.For oxidation prevention, sodium sulfite, sodium bisulfite, etc. As a coloring preventive agent, sodium thiosulfate, Rongalite, sodium carbonate, etc.
Citric acid, sodium citrate, glyalin, etc. can be used to prevent decomposition.

The surfactant is added to the large immunoglobulin aqueous solution containing the various additives mentioned above in order to suppress the formation of precipitates, and may be selected from among those with high safety, but in particular HLB11 -16 nonionic ones are desirable, and if these are used, the effect of suppressing precipitates in the preparation is remarkable. The appropriate amount of surfactant to be added is usually 0.02 to 3% (see Table 2).

Nonionic surfactants that can be used here include fatty acid polyhydric alcohol polyoxyethylene ether (ex, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil), alkyl polyoxyethylene ether, alkyl carbonyl polyoxyethylene,
Ethylene oxide adducts such as N,N=-di(polyoxyethylene)alkanamide, fatty acid sucrose ester, N,N''-di(alkanol)alkanamide, etc.
Examples include polyoxyethylene block copolymers.

In the above description, an aqueous solution was used as an example of the form of the eye drops, but other forms such as ointments and oils may of course be used.

<Actions and Effects of the Invention> The immunoglobulin preparation for eye drops of the present invention can be used to treat viral-induced epidemic keratoconjunctivitis and acute hemorrhagic disease, for which conventional eye drops containing antibiotics alone have not been able to provide sufficient therapeutic effects. conjunctivitis·
It can be expected to have a sufficient therapeutic effect on corneal herpes as well as severe infections caused by bacterial inflammation. In other words, by supplying immunoglobulin to the outer or inner eye via eye drops, if inflammation occurs in the cornea of the inner or outer eye, which has low immune function, a large amount of antigen will be released into the eye. It also prevents immunoglobulin from remaining in a localized state.
It is possible to prevent a decrease in immunoglobulin levels in the lachrymal fluid of patients with severe infections, regardless of the site of infection.

Further, by adding a nonionic surfactant with a specific HLB, the generation of precipitates is significantly suppressed, regardless of the type of auxiliary agent, as shown in the examples below, although the reason is unknown. Therefore, in this case, it is possible to arbitrarily select a buffer, an isotonizing agent, a thickener, a preservative, and a stabilizer to form an ideal ophthalmic globulin preparation.

<Example> Examples of the present invention will be described below.

A 2% human immunoglobulin aqueous solution having the formulation shown in Table 1 was stored in a cool, dark place (approximately 4°C), and the occurrence of precipitates (floating matter and precipitates) in the solution was observed for two weeks. The criteria for the judgment symbol are as follows (the same applies hereinafter).

Record number Floating matter/sedimentation - No floating objects or even a few floating objects.

+ A few floating objects.

+10 Lots of floating objects.

++10 Recognized as a precipitate.

+++10 Number of precipitates ~ turbidity.

In the formulations shown in Table 1, 0.1% of polyoxyethylene sorbitan monooleate was added to each, and the occurrence of precipitates was observed in the same manner as above. Even after a week had passed, the judgment was (-).

Furthermore, sample N092 human immunoglobulin aqueous solution or N
aCIG, 9% saline was instilled into the eyes of rabbits infected with epidemic keratoconjunctivitis (adenovirus type 8), and virus detection was performed for each group over time. As shown in Figure 1, globulin A clear difference in therapeutic effect was observed between the eye drop group and the saline eye drop group.

In addition, to the basic formulation of the immunoglobulin preparation shown below, various surfactants were added at various concentrations as shown in Table 2, further adjusted to pl (B, 7) with 0.5N hydrochloric acid, and filtered ( , 22 μm), and the generation of precipitates was observed in the same manner as above.

(Basic prescription) γ-globulin 2.0% sodium chloride 0.65% potassium chloride
0.16% calcium chloride
0.03% Sodium Bicarbonate 0
．． 003% Methyl paraoxybenzoate 0.02
%Propyl paraoxybenzoate 0.01% Hydrochloric acid
Appropriate amount of sterile purified water
- Amount: 100.0 The results are shown in Table 2. It can be seen that when a certain amount or less of a nonionic surfactant with HLB of 11 to 16 is added, there is a precipitate suppressing effect. On the other hand, in the case of cationic surfactants (benzalkonium chloride) and anionic surfactants (sodium lauryl sulfate), precipitation occurs immediately after adjustment, and no precipitate suppressing effect is observed. No (sample Nos. 33 and 34). In addition, if the nonionic surfactant has an HLB of less than 11, it will be difficult to dissolve in water and become cloudy (
Sample No.

17・26), conversely, nonionic surfactants have HLB16
If it exceeds , the lipophilicity becomes too small and the precipitate suppressing effect becomes small (Sample No. 24). The amount of nonionic surfactant added varies depending on the type of surfactant, but
In order to exhibit a precipitate suppressing effect, approximately 0.02% is required (sample No. 22/30).

In addition, in sample Nos. 21 and 30 of Table 2, for immunoglobulin preparations using pepsin-treated immunoglobulin and sulfonated immunoglobulin instead of large immunoglobulin, the buffer solution was replaced with boric acid, respectively. The following formulation I is a salt/phosphate buffer solution and the immunoglobulin is also denatured! ! When we observed the occurrence of precipitates for each of the immunoglobulin preparations in the same manner as described above, all of them were judged as (-) even after 2 weeks.
)Met.

In addition, in Formulation 11, 0.01% of benzalkonium chloride, a cationic surfactant, is added as a preservative, but with this amount, adding an appropriate amount of a nonionic surfactant can suppress precipitates. It does not affect the effect in any way.

(Prescription PEG-treated immunoglobulin 1.0% boric acid
1.0% boric acid
0.1% sodium chloride
0.35% polyoxyethylene hydrogenated castor oil 40 (HLB12.5) 3.0
% Thimerosal 0.02% Sterile, distillate 100.0% (Formulation I!) Alkylated human immunoglobulin 0.50% phosphoric acid
Sodium 0.25% Disodium Phosphate 0.25% Sodium Chloride
0.65% polyoxyethylene sorbitan trioleate ()ILB11) 0.1
% Benzalkonium chloride 0.01% Distilled water ゛ Amount 100.0% Furthermore, to the basic formulation, polyoxyethylene hydrogenated castor oil 60 and polyoxyethylene sorbitan monooleate were added at 0.02% and 0.05%.・0.1%・0.5%・
As a result of measuring the neutralizing antibody titer of large immunoglobulin for each of the immunoglobulin preparations to which 1%, 2%, 5%, and 10% were added, no inactivation or decrease in the neutralizing antibody titer was observed. Ta.

To determine the neutralizing antibody titer, a virus standard strain was used, a 2% solution of virus and large immunoglobulin was diluted with sterile distilled water, and an inactivation reaction was performed at room temperature. The reaction was stopped by dilution, and inactivation was determined by measuring residual infectious virus after the reaction using human fetal lung cells according to Black's method.

[Brief explanation of the drawing]

FIG. 1 is a graph showing the therapeutic effect when an immunoglobulin preparation is instilled into the eyes of a rabbit suffering from epidemic keratoconjunctiva. Patent application Fujirebio Co., Ltd. Japan Eye Drop Research Institute Co., Ltd. Agent

Claims (6)

[Claims] (1) An immunoglobulin preparation for eye drops, which contains immunoglobulin as an active ingredient and is applied to affected areas of bacterial or viral ophthalmological diseases. (2) At least one of an isotonizing agent, a buffering agent, a stabilizer, and a surfactant is appropriately added to the aqueous solution containing 0.1 to 15% of the immunoglobulin. The immunoglobulin preparation for eye drops according to item 1. (3) A patent claim characterized in that the immunoglobulin is a human immunoglobulin or one obtained by subjecting the human immunoglobulin to pepsin treatment, plasmin treatment, sulfonation treatment, polyethylene glycol treatment, acid treatment, or alkylation treatment. The immunoglobulin preparation for eye drops according to item 1. (4) The immunoglobulin preparation for eye drops according to claim 1, wherein the surfactant is a nonionic surfactant with an HLB of 11 to 16. (5) Claim 1, wherein the nonionic surfactant is polyoxyethylene hydrogenated castor oil.
The immunoglobulin preparation for eye drops described in Section 1. (6) The immunoglobulin preparation for eye drops according to claim 1, wherein the nonionic surfactant is polyoxyethylene sorbitan fatty acid ester.