JPS6261927A - Adjuvant for immuno-enhancing agent - Google Patents

Abstract

PURPOSE:To provide an adjuvant containing a substance selected from chitin oligomer, chitosan oligomer and partially deacetylated chitin oligomer as an active component and effective as an adjuvant when administered in combination with an antigen or as an immuno-enhancing agent when used singly. CONSTITUTION:The objective adjuvant of immuno-enhancing agent contains at least one kind of substance selected from chitin oligomer (e.g. di-N-acetyl- chitobiose), chitosan oligomer (e.g. chitobiose) and partially deacetylated chitin oligomer (e.g. di-N-acetyl-chitobiose) as an active component. Since the active component has essentially free from toxicity and has excelent immuno-enhancing activity and adjuvant activity to improve the liquid antibody-productivity and cellular immunity, it is expected to exhibit excellent adjuvant effect as an adjuvant to induce an antibody-producing cell in vaccination.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a substance that increases the immunological effect of an antigen when injected into a host together with an antigen in immunotherapy for various diseases, that is, an adjuvant. . The present invention also relates to immune enhancers that are administered alone.

(Prior Art) Conventional adjuvants include alum and aluminum phosphate as precipitating adjuvants, and mineral oils such as Payol F, Drakeol, Clearol, etc. as antibody-producing cell-inducing adjuvants. Oil-based adjuvants are mentioned (10th edition Japanese Pharmacopoeia Explanation, pages E-73Y to 1 Hiro O, published by Hirokawa Shoyo on February 7, 1929).

(Problems to be Solved by the Invention) However, the oil-based adjuvant used as an antibody-producing cell-inducing adjuvant is very poorly absorbed in the body after injection, and granulomas remain for a long time, resulting in However, the disadvantage is that the induration that appears as induration does not disappear for a long time, so only Myononan and aluminum phosphate have been put into practical use as precipitating adjuvants. It is hoped that a sexual adjuvant will emerge.

(Means for Solving Problems 1) In view of the current situation, the present inventors have conducted intensive research to provide a practical and novel antibody-producing cell-inducing adjuvant. Water-soluble chitin oryzamer (also called N-acetyl chitooligosaccharide) and chitosan oligomer (also called chitooligotang), which are known to be obtained by hydrolyzing chitin.
and that partially deacetylated chitin oligomers are surprisingly useful as immune enhancers when administered alone;
They also discovered that it has excellent adjuvant activity when administered together with an antigen, leading to the completion of the present invention.

The active ingredients of the adjuvant or immune enhancer of the present invention are:
It is made of at least one substance selected from chitin oligomer, chitosan oligomer, and partially deacetylated chitin oligomer. Specific examples of chitin oligomers include di-N-7 cetyl-chitobiose, tri-N
-acetyl-chitotriose, tetra-N-acetyl-
Chitotetraose, penta-N-acetyl-chitohentaose, hexa-N-acetyl-chitohexaose.

Hepta-N-acetyl-chitoheptaose,
Specific examples of chitosan oligomers include chitobiose, chitotriose, chitopentaose, chitohexaose,
Examples of partially deacetylated chitin oligomers include di-N-acetyl-chitobiose, tri-N-acetyl-chitotriose, tetra-N-acetyl-chitotetraose, and penta-N-
Acetyl-chitopentaose, hexa-N-acetyl-
Examples include chitohexaose and penta-N-acetyl-chitopentaose from which a portion of the acetyl group has been removed.

The adjuvant of the present invention can be used in combination with various immunogenic antigens, such as vaccine preparations, to subcutaneously, subcutaneously, intramuscularly, or
Administer by intraperitoneal, intracavitary or oral administration. Also 1
When the active ingredient substance of the present invention is administered alone, it acts as an immune enhancer. On the 7th day, the effective dose per gram of body weight is /~0OIII+, preferably ! of at least one substance selected from chitin oligomers, chitosan oligomers, and partially deacetylated chitin oligomers.
It is 0 to ISO Ki.

(Effects of the present invention) The active ingredients of the adjuvant or immune enhancer of the present invention are chitin oligomers obtained by hydrolyzing naturally occurring chitin, chitosan oligomers, and partially deacetylated chitin oligomers. Since it consists of at least one substance IX and has immunoenhancing activity and adjuvant activity to enhance cellular immunity, antibody production aJ
It is expected to have an excellent adjuvant effect in vaccine administration as a cell-inducing injection adjuvant.

(Example) Formulation Nl/Preparation of injection Hexa-N-acetyl-chitohexaose/Q? was dissolved in an appropriate amount of physiological saline for injection to make an aqueous solution with a total strength of 7,000, and an injection was obtained according to the method specified in the 10th Pharmacopoeia for injections.

Formulation Example 2 Preparation of Injectable Chitohexaose 10f was dissolved in an appropriate amount of physiological saline for injection to make an aqueous solution with a total volume of 1000, and an injection was obtained according to Formulation Example.

Next, the antibody production enhancing effect and fine foam immunity enhancing effect of the adjuvant or immune enhancer of the present invention will be illustrated by experimental examples.

Actual example/Antibody production enhancement test Hexa-N-acetyl-chitohexaose (test Nll/) or chitohexaose prepared in Formulation example/and Formulation example λ was administered into the tail vein of BALB10 male mice aged ψ to 6 weeks. An injection solution containing (Test m2) as an active ingredient was administered 3 times in total for 3 consecutive days at a dose of 00 HI kg of mouse per dose of active ingredient. One day later, sheep erythrocytes were administered to each mouse as an antigen, and the lungs were removed and the lung cells were transformed into Jerne's plaque-forming cells (plaque-forming cells).
The number of antibody-producing cells per lung was determined by a measuring method (que forming cell), and the effect of enhancing antibody production was investigated. The control test was conducted in the same manner as described above, except that only hexa-N-acetyl-chitohexaose and physiological saline containing no chitohexaose were used. The results obtained are shown in Table-/.

Table-/Efficacy of enhancing antibody production ability of mouse lung cells/)
The number of antibody-producing cells/lung shows the mean difference between mice. 7'& 2) I) indicates a risk rate of 7% or less by 5student's t-test.

Test Example 2 Cellular Immunity Enhancement Test Hexa-N-acetyl-kitohexaose (Test N[L3) or chitohexaose (Test N [L3)] or chitohexaose (Test N[L3) prepared in the formulation Reli/and backing example Co.
An injection containing 10 Owy/kit mice of the active ingredient was administered 3 times in total for 3 consecutive days each time. Seven days later, 1×10 sheep red blood cells were administered per mouse as an antigen.

After that day, sheep red blood cells were again administered as an antigen at 108 cells/mouse, and 7 days later, the swelling in the front of the legs was measured to determine the rate of increase in edema, and the effect of enhancing cell-mediated immunity was examined. The control test was conducted in the same manner as described above, except that only hexa-N-acetyl-chitohexaose and physiological saline containing no chitohexaose were used. Table 2 shows the results obtained.
Shown below.

Table-1 Cell-mediated immunity enhancement effect test in mice/ha/
Significant differences were observed for X and Na2 at 5% or less.

Procedural amendment October 7, 1985

Claims (1)

[Claims] An adjuvant or immune enhancer containing as an active ingredient at least one substance selected from chitin oligomers, chitosan oligomers, and partially deacetylated chitin oligomers.