JPS5892619A - Stable composition containing interferon - Google Patents

Abstract

PURPOSE:To provide the titled composition containing interferon as active component and a specific sugar alcohol as stabilizing agent. CONSTITUTION:The objective composition containing stably compounded interferon is prepared by compounding >=1X10<4> IU of interferon (having a titer of >=1X10<5> IU/mg-protein) per 100g of the composition and >=15wt%, preferably >=35wt% stabilizer comprising >=3-hydric sugar alcohol (e.g. glycerin, erythritol, arabitol, etc.) based on the whole composition. The stability of interferon can be further improved by adding an acidic sugar, an organic acid buffering agent, a sulfur-containing mild reducing agent having sulfuhydryl selectivity, a human serum albumin, etc. to the composition. It can be administered in the oral cavity, to the skin, or in the rectum, vagina, urethra, eye, ear, or nose, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to compositions stably containing interferon.

Interferon is a type of protein that is produced by animal cells in response to stimulation by viruses, double-stranded RNA, etc. and has the effect of suppressing the proliferation of viruses, and its effect is known to be specific to animal species.

In recent years, it has become increasingly clear that interferons derived from human cells and microorganisms incorporating human interferon genes exhibit therapeutic effects on various human diseases, and clinical applications are being attempted.

Because of the therapeutic effects of interferon, it is expected that interferon can be used as a medicinal agent to be incorporated into various compositions including pharmaceuticals, quasi-drugs, and cosmetics. However, interferon is an extremely unstable substance, and in particular, when it is purified to a clinically applicable level, its activity is significantly reduced by temperature (high temperature) or physical pressure, and it easily expires. However, unless the interferon is stabilized in the composition, the effects of its combination cannot be expected.

Therefore, the present inventors continued their intensive research in order to obtain a composition containing human-derived interferon in a stable manner, and found that, unexpectedly, when a certain sugar alcohol is present in the composition, the amount of interferon in the composition increases. It was discovered that the stability was significantly improved, and the present invention was completed.

That is, the present invention provides a composition stably containing interferon, which contains human-derived interferon as an active ingredient and contains a trivalent or higher valent sugar alcohol as a stabilizer. According to the present invention, the activity of interferon can be improved even if interferon is incorporated into various compact compositions for oral administration, external skin administration, rectal, vaginal or intraurethral administration, ocular, ear or intraepinal administration. is maintained stably over a long period of time.

Thus, the interferon incorporated in the composition of the present invention may be of any human origin, for example, human leukocytes, normal diploid cells, or microorganisms into which human interferon genes have been incorporated using recombinant DNA techniques. are used. The blending amount is not particularly limited and can be selected appropriately depending on the actual shape, etc., but from the viewpoint of effectiveness, it is generally lXIO3 international unit /
Composition 1 contains interferon with a titer higher than da protein.
It is preferable to mix it at a ratio of 1X10' international unit or more per 00g.

Examples of trihydric or higher sugar alcohols used as stabilizers include glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol, etc.
These may be used alone or in combination of two or more, and from the viewpoint of stabilizing effect on interferon, the blending amount is 15% (wt%) based on the total amount of the composition as a sugar alcohol.
The same applies hereinafter) or more, preferably 85% or more.

As mentioned above, the composition of the present invention can be used in small forms for oral application such as pasta, toothpaste, and mouthwash, in small forms for external use on the skin such as gels and ointments, and for rectal and vaginal applications such as gels and mixtures. It can be in a variety of forms, including forms for intraurethral administration, forms for ocular, otic or intranasal administration such as solutions, gels, ointments, and sprays. These can be manufactured according to conventional formulation techniques, and interferon may be added in the final step of formulation. Other ingredients are not particularly limited as long as they do not affect the stability of interferon, and any of those commonly used in this type of preparation may be used. In addition, along with trivalent or higher sugar alcohols, iduronic acid, galacturon 111, ri) V-curonic acid,
Currona, mannuronic acid, ketogluconic acid, ketogluconic acid, ketogulonic acid, ascorbic acid, etc. are sugars and their salts, citrate buffers, succinate buffers,
Organic acid buffers such as tartrate buffers, fumarate buffers, gluconate buffers, oxalate buffers, lactate buffers, acetate buffers, thioctic acid, N-acetylcysteine, N- Sulfhydryl group-selective mild sulfur-containing reducing agents such as acetylhomocysteine, glutathione, thiodiglycol, thioethanolamine, monothioglycerol, dithiothreitol, thioalkanoic acids having 1 to 7 carbon atoms, and human serum albumin. Since the stability of interferon is further improved by blending one or more substances selected from the following, these may be blended as appropriate.

Next, the present invention will be described and explained in more detail with reference to actual examples and examples. The interferon used in these experimental examples and examples was obtained from human foreskin-derived fibroblasts, and its activity was measured using Sindbis virus (3indbi 5virus) and human amnion-derived cell lines. Cells (FL cells) were measured by the cytopathic effect (CPE) method.
l, E, A, and Vi lεek, J
(1972).

Antimicrob, Agents Chemot
her, 2, 476; Oie, H, K, (1
977), Texas Rep, 'Bio1. Med
, , 85154). Furthermore, the interferon titer was compared with the activity of standard interferon measured at the same time and converted into international units (hereinafter abbreviated as IU).

Experimental Example 1 Lyophilized interferon I
An interferon solution with a concentration of 105 IU/- was prepared. 01 l of this solution was mixed with 0.9 l of physiological saline solution containing various sugar alcohols and heated at 45°C for 24 hours.
I left it for a while. After 24 hours, the interferon activity of each mixture was measured, and the initial activity (I
The interferon activity retention rate was calculated using U/x, t ) as 100%.

Table 1 shows the relationship between the type and concentration of the sugar alcohol used and the residual activity rate.

Experimental Example 2 Same as Experimental Example 1, left at 37℃ for 1 month and 4℃
I left it alone for 6 months. The results are shown in Table 2.

Table 2 As is clear from the results in Tables 1 and 2, all sugar alcohols have interferon stabilizing effects;
In particular, glycerin, xylitol, and erythritol have shown excellent effects.

Example 1 Pasta was prepared according to a conventional method using the following recipe.

Pasta prescription ingredients % Setanol 2.0 glycerin! Lemonostearate 9.27 Tween 80 2.0 Hydroxyethylcellulose 5.5 Caliy
0.09 sugar alcohol (shown in Table 3) 40 Purified water The remaining components were mixed, interferon was added at a ratio of 1X10IU/100y pasta, and the mixture was thoroughly mixed to obtain interferon self-compound pasta.

After storing the obtained pasta at 37°C for 1 month and at 4°C for 6 months, its interferon activity was measured, and the residual rate of interferon activity was calculated by setting the activity immediately after production as 100%. The results are shown in Table 3.

Table 8 Example 2 A toothpaste was prepared according to the conventional method according to the formulation shown in Table 4. After mixing each component, finally add 1X1 interferon.
The mixture was added at a ratio of 0'IU/100N toothpaste and mixed well to obtain an interferon-containing toothpaste.

This was prepared at 37°C for 1 hour in the same manner as in Example 1 for the pasta.
After storage for 6 months and 6 months at 4°C, the residual activity of the interferon was calculated. The results are shown in Table 4.

Table 4 Example 8 A gel preparation for external use on the skin was prepared according to a conventional method using the following formulation.

Gel formulation ingredients % Sugar alcohol (shown in Table 5) 45 Tween 3Q O,2 Carbopol 940 2 Purified water
The remaining components were mixed, and finally interferon was added and mixed at a ratio of IX to 7 IU/100 f gel to obtain an interferon-containing gel for external use on the skin.

This was heated to 87°C in the same way as in the pasta of Example 1.
After storage for 1 month and 6 months at 4°C, the residual activity of interferon was calculated. The results are shown in Table 5.

Table 5 Example 4 A gel preparation for external use on the skin was prepared according to a conventional method using the following formulation.

Gel formulation ingredient % Erythrit-I 45
Carbopol 9-40 2 Purified water Mix the remaining ingredients,
Finally, add interferon lX106IU/100g
were added and mixed in the following proportions to obtain an interferon-containing gel for external use on the skin. Apply this gel at 37°C for 1 month.
! The residual activity rate of interferon when stored at 4°C for 6 months was 30% and 45%, respectively. In addition,
As a control, gels obtained by replacing erythritol with purified water were similarly subjected to a storage test, and the residual activity rates were 0% and 1%, respectively.

Patent applicant Sunstar Co., Ltd. and 1 other patent attorney Aoyama Naked wire or 2 people

Claims (1)

[Scope of Claims] (1) A composition stably containing interferon, which contains interferon as an active ingredient and contains a sugar alcohol of octavalent or higher valence as a stabilizer. (2) The composition according to item (1) above, wherein the sugar alcohol is one or more sugar alcohols selected from the group consisting of glycerin, erythritol, arabitol, xylitol, sorbitol, and mannitol. (8) The composition according to item (1) above, which contains 15% by weight or more of a sugar alcohol based on the total amount of the composition. (4) Furthermore, as a stabilizer, acidic sugar, organic acid buffer,
The composition according to any one of items (1) to (8) above, which contains one or more substances selected from the group consisting of a sulfhydryl group-selective mild sulfur-containing reducing agent and human serum albumin. . (5) Items (1) to (4) above, which are narrow shapes for intraoral application.
) The composition of any one of the items. (6) Items (1) to Water (4), which are thin forms of skin external preparations.
) The composition of any one of the items. (7) Paragraph (1) of llJ, which is a narrow form for intrarectal administration.
The composition of any one of paragraph (4). (8) Items (1) to (4) above, which are in a narrow form for intravaginal administration.
) The composition of any one of the items. (9) Items (1) to (1) above, which are in a narrow form for intraurethral administration.
4) The composition of any one of item 4). (lO) Items (1) to (4) above in a narrow form for ocular administration.
) The composition of any one of the items. (11) The above-mentioned items (1) to (4) are in a narrow form for ear administration.
) The composition of any one of the items. (12) Items (1) to (4) above, which are narrow for nasal administration.
) The composition of any one of the items.