JPS591691B2 - Intravenous γ-globulin preparation - Google Patents

Description

[Detailed description of the invention] The present invention relates to intravenous gamma-globulin preparations.

More specifically, the present invention relates to an intravenous γ-globulin preparation containing γ-globulin and a selected dissociating agent.

Among immunoglobulins, which are plasma protein components, γ-globulin preparations, which mainly consist of IgG, have been widely used for the prevention and treatment of various infectious diseases, but due to the high anti-complement value of these preparations, , was limited to intramuscular administration.

However, due to the high medical demand for this preparation, we are hoping for more efficient, larger-scale administration, and faster efficacy.
Attempts have been made to apply this to intravenous administration, and several methods have been proposed.

In order to enable intravenous administration of γ-globulin preparations,
It is necessary to reduce anti-complement levels, which are the cause of side effects.
It has now been confirmed that the anti-complement value of γ-globulin is caused by aggregation of its monomers during the fractionation process of γ-globulin.

Several methods have been proposed to reduce this anti-complement titer, including the following.

(1) Enzyme treatment method [Deutsche Medizini
87.19
43 (1962), Fox Sanguinis (Vox
Sanguinis) 13.71-84 (196
7), l.

(2) Chemical modification method (Fox Sanguinis 28°4
22-437 (1975).

(3) Method for separating and removing aggregates from γ-globulin preparations [Fox Sanguinis 7.157-174
(1962), JP-A-49-101516, JP-A-49-81519].

(4) Method for separating aggregates contained in γ-globulin preparations into monomers [Acta Chemi-Ryoku Scandinapica (A
cta Chemica 5candinavic
a) 22°490-496 (1968)).

However, method (1) shown here, in which immunoglobulin is treated with pepsin to cleave the complement-fixing moiety from the globulin molecule (published in the German Chemedeichen Societies Wotzchenschrift), Even if the complement value is lowered, it has the disadvantage that the duration of the active portion of the antibody in the blood is significantly shortened, and in addition, if pepsin is included in the preparation, it may cause antigenic disease.

The method of treating immunoglobulin with human plasmin and cleaving globulin aggregates into monomers (Fox Sanguinis Shellfish, 71-84) does not cause problems with antigenic diseases;
Although this is an excellent method, it cannot be said to be a perfect method since a portion of the monomer is cleaved to form a truncated small molecule, which lasts only a short time in the blood.

The method (2) has the disadvantage that it differs from the natural state of γ-globulin.

Methods (3) and (4) are the most desirable in terms of obtaining γ-globulin in its natural state, but these methods also produce aggregates during storage and the anti-complement value decreases. The disadvantage of rising remained.

Therefore, the present inventors discovered that γ-globulin in its natural state,
Moreover, by creating γ-globulin with a low anti-complement value in advance using a known method, researching a method to stabilize it for a long period of time, and adding γ-globulin selected from glucose, glycine, and sodium chloride to a relatively high concentration. He discovered that it could be dramatically stabilized and filed a patent application on October 13, 1976 (
Patent Application No. 122570/1970) was filed.

The present inventors further continued their research and found that there are substances other than these stabilizers that exhibit a stabilizing effect, thereby completing the present invention.

That is, the present invention allows medical γ-globulin to contain at a relatively high concentration one or a mixture of two or more selected from fructose, galactose, arabinose, ribose, maltose, and lactose as a substance that dissolves aggregates. The present invention consists of providing a γ-globulin preparation suitable for intravenous injection with a low anti-complement titer.

The human-derived γ-globulin used in the present invention does not need to have a low anti-complement value as long as it is in its natural state.
γ that can be obtained by any method, but can be manufactured using existing equipment and is already used as a medicine for intramuscular injection.
- It is most efficient to use globulins.

The concentration of γ-globulin added in the formulation is between 5 and 20
% w/v is preferred.

This concentration is determined from the amount of dissociating agent added and the dilution factor at the time of administration to the living body.

As the saccharide used as a dissociating agent, monosaccharides such as fructose, galactose, arabinose, and ribose, and trisaccharides such as maltose and lactose are effective, but those having properties as reducing sugars are particularly preferred.

The concentration is greater than or equal to 5% w/v concentration, more preferably 10
~20% w/v, or 40-100% saturation concentration amount.

In addition, when using these dissociating agents together, the total amount is 5 to 2.
As long as it contains a concentration of 0% w/v, there is sufficient effect.

After adding the dissociation agent, it is stored at 4 to 50°C for 2 to 30 days.

Preferably at 4°C for 20-30 days or at 37°C for 3-7 days.
It is best to store it for several days.

Note that when γ-globulin containing no aggregates is used as a raw material, it may be possible to formulate the product directly after adding a dissociating agent.

The preparation thus obtained is in the form of a solution, dissolved in water or a suitable buffer.

This pH is preferably 6.4 to 7.4.

After sterile filtration if necessary, the preparation is dispensed according to the packaging unit so as to contain 500-10,000 η of γ-globulin.

The storage method is not particularly limited as long as high temperatures are avoided, but it is particularly desirable to store it at 30°C or below, but it may also be a freeze-dried preparation.

When using this preparation, it is diluted or dissolved in distilled water for injection and administered intravenously.

The dilution ratio is preferably about 5 to 15 times.

The usual dosage for adults is 500-3000η of γ-globulin at one time, and for children, 250-1500η of globulin at one time.

As a safety test, an acute toxicity experiment was conducted.

Maltose 10 in a 10% solution of globulin for intramuscular injection (commercially available)
% and reacted aseptically at 37°C for 5 days,
Diluted 10 times with sterile distilled water and injected into the tail vein of a mouse in a total volume of Q, 5 ml, 1. Orut/mouse was administered to 5 animals per group and observed for 7 days, but no abnormality was observed.

The intravenous γ-globulin preparation according to the present invention thus obtained does not contain γ-globulin that has undergone enzymatic degradation or chemical modification.

Since it consists essentially of a form that exists in nature, it has a long half-life in the blood and is an extremely effective intravenous γ-globulin preparation without antigenicity problems.

In addition, any unmodified γ-globulin that can be obtained by any existing method can be used as a material for this preparation, and the aggregates contained therein can be removed or enzymatically cleaved as before. It is economically advantageous because it is dissociated into naturally occurring 7SIgG and used in its entirety without being fragmented, making it extremely desirable industrially.

This preparation has high storage stability and no increase in anti-complement value during long-term storage, indicating that it is medically extremely safe and highly effective as an intravenous γ-globulin preparation. I can say that.

In the present invention, the measles antibody titer is determined using the Hemagglutination Inhibition Test (Hemagglutination Inhibition Test).
It is measured using the international unit (IU) method.
/100■).

The anti-complement titer was measured using the method of Kabat and Mayer [Experimental Immunochemistry].
tal Immunochemistry), 22
5, (1961)) and the method of Saigoku and Ryogoku [Immunological Biochemistry, 103, 1972 (Kyoritsu Shuppan)].

That is, the number of units reduced by adding the sample from 100 units of complement was measured, and the unit of reduction was expressed as the anti-complement value.

Examples of the formulation of the present invention are shown below.

Example 1 Measles antibody titer is 9■U/100rnJ? The anti-complement titer was 67.
Dissolve 13 g of maltose (Kishida Chemical) in 100 ml of a 10% solution of γ-globulin for intramuscular injection showing 20 to 40 g.
The reaction was carried out aseptically at ℃ for 5 days.

After the reaction, the anti-complement titer decreased to 16, and the measles antibody titer at that time was 9 IU/100~, with no loss at all.

This product is aseptically dispensed as a preparation and once diluted with sterile distilled water before use.

Example 2 Measles antibody titer 8.5I obtained from placenta by ammonium sulfate fractionation method
Add 15 g of fructose (Kishida Chemical) to a 12% solution of placental globulin with an anti-complement value of 58 at U/100 m9 and add 10 to 2
The reaction was carried out aseptically at 5°C for 10 days.

After reaction, the anti-complement titer was 18 and the measles antibody titer was 8.6 IU/1.
It was 00■.

This product is aseptically dispensed as a preparation and used by diluting it once with sterile distilled water.

Example 3 7 g of maltose and 21 ribose were added to 100 U of a 10% solution of γ-globulin for intramuscular injection showing a measles antibody titer of 8 I [J/1007 V and an anti-complement titer of 35, and the mixture was sterilized at 30°C for 5 days. It was preserved.

After the treatment, sterilization was carried out through a Millipore filter, the P solution was dispensed, and freeze-drying was performed.

Comparative Experiment Example Additives (dissociators) to 10% γ-globulin solution 1001711, which has a measles antibody titer of 9 IU/100 ■ and an anti-complement titer of 67, include glucose, fructose, galactose, arabinose, ribose, maltose, lactose, glycine,
Alanine, sodium chloride, potassium chloride, and magnesium chloride were added alone or in combination (the amounts added are specified in the table), and the antibody titer of each test solution (IU/100 ) The anti-complement value was measured and the effects of each additive were compared.

Furthermore, each test solution and each lyophilized product were stored at room temperature for 2 months, and their stability over time was also compared.

The results are shown in the table, and the compositions of the present invention showed significant differences in effectiveness in all cases.

Claims (1)

[Claims] 1 γ-globulin manufactured for medical use 5-20% W/
A γ-globulin preparation for intravenous injection or lyophilization thereof, characterized in that V contains a saccharide selected from fructose, galactose, arabinose, ribose, maltose, and lactose, alone or in combination, in a total amount of 5 to 20% w/v. formulation.