JPH07258097A - Agent for suppressing excessive production of tnf - Google Patents

Abstract

PURPOSE:To provide an agent for suppressing excessive production of TNF, containing a specific nucleic acid component such as inosine and cytidine as an active component, effective for suppressing the excessive production of TNF by the administration to a patient of inflammatory diseases, etc., and exhibiting excellent effect for the prevention and treatment of systemic inflammation reaction, etc. CONSTITUTION:This suppressing agent contains (A) inosine, (B) cytidine, (C) uridine, (D) guanosine-5'-monophosphate or its salt and (E) thymidine at molar ratios of preferably 4:4:3:4:1. The component D is preferably Na salt, concretely guanosine-5'monophosphate disodium salt, etc.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a TNF overproduction inhibitor,
More specifically, it relates to the above TNF overproduction inhibitor containing a specific nucleic acid component such as inosine and cytidine as an active ingredient.

[0002]

[Prior art and its problems] TNF (tumor necrosis facto)
(R; tumor necrosis factor) is a kind of cytokine secreted mainly from macrophages, and when it is secreted in a normal amount into the living body, it enhances the immune capacity of the living body [Ghiar
a, P., Boraschi, D., Neucioni, L., et al., J. Immuno
l., 139 , 3676-3679 (1987)], for example, Listeria monocytogenes,
Although it has a protective effect against infection by Legionella bacteria, Mycobacterium bacteria, etc., and is beneficial to the living body, if the above TNF is overproduced or secreted for some reason,
The body has systemic inflammatory reaction symptoms [Members of the American
College of Chest Physician Society of Critical Car
e Medicine Consensus Conference Commitee, Crit. Ca
re Med., 20 , 864-873 (1992)], anorexia, catabolism, weight loss, anemia and other pathological conditions are known to occur. From this aspect, TNF is also considered to be a factor inducing inflammation and the like. To be

[0003] Therefore, various studies have hitherto been made for the above-mentioned TNF to suppress its overproduction or to prevent or suppress the action of the excessively produced TNF as the above-mentioned inducing factor. For example, research on monoclonal antibodies against TNF is one of them (Smith, SR, Calzet
ta, A., Bankowski, J., et al., J. Leukoc. Biol., 54 ,
23-29, 1993).

However, such a monoclonal antibody against TNF is not only expensive per se, but its application to a living body also reduces the beneficial effects of TNF such as biological defense. There are drawbacks.

[0005] In addition to the above-mentioned antibodies, substances that have the effect of suppressing TNF overproduction and are considered to be useful as pharmaceuticals have not yet been proposed, and the development of a drug capable of suppressing such TNF overproduction is in the field. It is in the current state of demand.

Therefore, an object of the present invention is to provide a TNF overproduction inhibitor which meets the above-mentioned needs of the art.

[0007]

As a result of intensive studies, the present inventors have found that the above object can be achieved by the combination of the following specific nucleic acid constituents, and have completed the present invention. It was

[0008] That is, according to the present invention, a TNF overproduction inhibitor characterized by containing inosine, cytidine, uridine, guanosine-5'-1 phosphate or a salt thereof and thymidine as an active ingredient, particularly inosine. , Cytidine,
There is provided the above TNF overproduction inhibitor, which comprises uridine, guanosine-5'-1 phosphate or a salt thereof and thymidine in a molar ratio of 4: 4: 3: 4: 1.

The above-mentioned guanosine-5'-1 phosphate salts include various conventional pharmacologically acceptable salts such as sodium salts and potassium salts, and among them, those having particularly high solubility are known. The sodium salt is preferred.

The nucleic acid constituent composition constituting the TNF overproduction inhibitor of the present invention is one that the applicant of the present invention has been engaged in research for a long time, and as described in, for example, Japanese Patent Publication No. 5-34337. It is known. However, although the publication describes that the composition has a nutritional supplementation effect, there is no basis for suggesting that the composition has a medicinal effect of suppressing TNF overproduction, and this fact indicates that the present invention It was first discovered by.

The TNF overproduction inhibitor of the present invention can be easily prepared by simply combining and appropriately mixing the nucleic acid constituent components as the above-mentioned active ingredients, but generally, the administration route, administration method and the like are It is shaped or prepared into an appropriate dosage form (dosage unit form) accordingly. Examples of the dosage unit form include liquid forms such as injections suitable for intravenous administration and oral administration,
Examples thereof include powders, tablets, pills, granules, powders, solutions, suspensions, emulsions, capsules and the like, which are suitable for tube administration, topical administration and the like. Preparation of such a dosage unit form can be carried out using an appropriate pharmaceutical carrier according to a conventional method. Examples of the carrier for the preparation include diluents and excipients such as a filler, a filler, a binder, a moisturizer, a disintegrator, a surface active agent, and a lubricant, which are usually used, depending on the use form of the preparation. it can.

More specifically, the drug of the present invention in the form of an injectable solution containing, for example, an infusion solution is typically dissolved in distilled water for injection by mixing and dissolving a predetermined amount of each of the above components in the same manner as an ordinary injectable solution.
If necessary, various additives commonly used, such as hydrochloric acid, acetic acid, lactic acid, malic acid, citric acid, sodium hydroxide, potassium hydroxide, etc. In addition, the resulting aqueous solution is prepared by heat sterilization or sterile filtration.

The thus-prepared liquid preparation of the present invention has a pH similar to that of an injectable solution such as a normal infusion solution, generally in the range of about 3.0 to 9.0, preferably about 5.0 to 8.0. Can have a pH of. The concentration of the active ingredient in the drug is usually about 0.5 to 10 w / v%, preferably about 2 to 8 w / v%.

Further, the preparation of the present invention can be prepared in a solid dosage form such as tablets. For example, tablets are carriers such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose and silica. Excipients such as acids and potassium phosphates; water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, polyvinylpyrrolidone and other binders; sodium carboxymethylcellulose,
Carboxymethyl cellulose calcium, low-substituted hydroxypropyl cellulose, dry starch, sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, and other disintegrants; polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, etc. Surfactants; sucrose, stearin, cocoa butter, hydrogenated oils, etc. disintegration inhibitors; quaternary ammonium base, absorption promoters such as sodium lauryl sulfate; glycerin, starch and other moisturizers, starch, lactose, kaolin, bentonite Adsorbents such as colloidal silicic acid; lubricants such as purified talc, stearates, boric acid powders, polyethylene glycols, etc. can be used and prepared by a conventional method. Further, the tablets are tablets coated with a usual coating as necessary, for example, sugar-coated tablets, gelatin-coated tablets,
It may be an enteric coated tablet, a film coated tablet, a double tablet or a multilayer tablet.

In the case of molding in the form of pills, as a pharmaceutical carrier, for example, glucose, lactose, starch, cocoa butter,
Excipients such as hardened vegetable oil, kaolin and talc, gum arabic powder, tragacanth powder, binders such as gelatin and ethanol, disintegrating agents such as laminaran and agar can be used.
The powder is obtained by sieving the active ingredient crystals, which have been sieved with a sieve, with lactose, sucrose, sodium chloride, glucose, as a carrier (excipient),
It can be prepared by uniformly mixing with urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid and the like. Capsules are usually hard gelatin capsules prepared by mixing the active ingredients of the present invention with the various pharmaceutical carriers exemplified above according to a conventional method,
It can be prepared by filling a soft capsule or the like. Furthermore, in the drug of the present invention, a colorant, a preservative, a fragrance, a flavoring agent, a sweetening agent and the like and other medicinal products can be contained, if necessary.

When preparing the preparation of the present invention in the form of an infusion solution, the preparation of the present invention in the form of the infusion solution contains sugars such as glucose, fructose, xylitol, sorbitol and maltose, lipids, vitamins and electrolytes. , Trace elements, etc. can be added and blended, and if necessary, a stabilizer, p
It is also possible to add and blend an H regulator and the like.

The administration method of the above-mentioned preparation of the present invention is not particularly limited, and it is determined according to various preparation forms, patient's age, sex and other conditions, degree of disease and the like. For example, tablets, pills, solutions, suspensions, emulsions, granules, capsules and the like are orally administered, and injections are intravenously administered alone or mixed with a normal replenisher solution such as glucose and amino acid, and further required. Independently, it is administered intramuscularly, intracutaneously, subcutaneously or intraperitoneally.

The dosage of the preparation of the present invention is appropriately selected depending on the usage, age of the patient, sex and other conditions, degree of disease and the like, but usually the amount of the active ingredient is about 0.
An amount of about 5 to 5.0 g, preferably about 1.5 to 2.5 g can be set as a guide, and this amount can be appropriately increased or decreased. The above-mentioned preparation can be administered in 1 to 4 divided doses per day.

[0019]

INDUSTRIAL APPLICABILITY The TNF overproduction inhibitor of the present invention suppresses the overproduction by administering it to a patient in a state where TNF is excessively produced in the body, for example, a patient with surgical invasion or infection. Thus, it is possible to exert an excellent effect on prevention and treatment of systemic inflammatory reaction and the like.

[0020]

EXAMPLES In order to explain the present invention in more detail, examples of production of the TNF overproduction inhibitor of the present invention will be given below, and then examples of pharmacological tests conducted on the active ingredient of the present invention will be given.
In addition,% in each example shows w / v%, and the molar ratio shows an approximate value. GMP · 2Na is guanosine-5′-1 phosphate disodium salt.

[0021]

[Production Example 1] Inosine 1.06%, cytidine 1.46
%, GMP · 2Na 2.44%, uridine 1.10%,
Thymidine 0.36%, purified sucrose 20.00%, ethyl paraoxybenzoate 0.009% and butyl paraoxybenzoate 0.006% Weigh each component so as to have a composition, and first heat purified water, Purified sucrose as a sweetness amount was added to this and dissolved by stirring, and after cooling, each nucleic acid constituent component, ethyl paraoxybenzoate and butyl paraoxybenzoate as a preservative dissolved in a small amount of ethanol were added and dissolved by stirring. . Then, after adjusting the liquid volume with purified water, it is filtered,
After filling a glass container and purging with nitrogen, the container was closed and heat sterilized to prepare a TNF overproduction inhibitor of the present invention in a liquid formulation.

Inosine: Cytidine: GMP
The molar ratio of 2Na: uridine: thymidine is about 4: 4 :.
It was 4: 3: 1 and the total free nucleic acid concentration was 6.8%.

[0023]

[Production Example 2] Inosine 2.4 g, cytidine 2.2 g, GMP · 2Na 3.7 as pure crystals of each nucleic acid constituent
g, 1.7 g of uridine and 0.5 g of thymidine, each of which was sieved through a 60-mesh sieve and mixed, then 89.5 g of starch was added as an excipient and mixed evenly, and filled in a glass container. Then, a powdered form of the TNF overproduction inhibitor of the present invention was prepared. The molar ratio of inosine: cytidine: GMP.2Na: uridine: thymidine of this product is
It was about 4: 4: 4: 3: 1.

[0024]

[Test Example 1] Preventive effect test for disorders caused by overproduction of TNF 20 Wistar male rats (7 weeks old) were divided into 4 groups, each of which was cannulated in the jugular vein for blood collection, and then metabolized for 24 hours. They were allowed to freely eat and drink water in the cage.

15 ml of physiological saline was added to one group of rats.
/ Kg intraperitoneally, and 60 minutes after that, lipopolysaccharide (LPS, manufactured by Difco, E. coli O111: B4) was used.
5 mg / kg was intraperitoneally administered.

Rats in the second group were treated with physiological saline in the same manner as in the first group, and 60 minutes after the administration, LPS was intraperitoneally administered at 10 mg / kg.

In the 3 groups of rats, a nucleic acid component composition (inosine: cytidine: uridine: GMP · 2Na: thymidine = 4:
4: 3: 4: 1 (molar ratio)) 15 ml / kg
Administered intraperitoneally, and 60 minutes later LPS 2.5 mg / k
g intraperitoneal administration.

The rats of the 4 groups were intraperitoneally administered with 15 ml / kg of the nucleic acid component composition as the active ingredient of the inhibitor of the present invention, and 60 minutes after that, LPS was intraperitoneally administered with 10 mg / kg.

For each group of rats, immediately before LPS administration and 30 minutes, 60 minutes, 90 minutes and 120 minutes after LPS administration
Minutes and 180 minutes, blood was collected from a jugular cannula, plasma was separated from the blood, and the amount of TNF-α in the plasma was measured by EL.
ISA (Enzyme linked immunosorbent assay, Parki
nson et al., J. Immunol. Methods, 15 , 105, 1988)
Was measured according to.

FIG. 1 shows the time course of plasma TNF of rats in each group.
(Horizontal axis = time (minutes), vertical axis = plasma TNF amount (pg / m
l)).

In the figure, (1) shows one group, (2) shows two groups,
(3) shows 3 groups, (4) shows 4 groups, respectively. Further, in the figure, the results of the significance test (by Tukey-test) of the values of the 4 groups against the values of the 2 groups as a control are shown by the following symbols.

* ... p <0.01, ** ... p <0.05 According to the figure, the plasma TNF amount shows a peak 90 minutes after LPS administration in each group, but the composition of the active ingredient of the TNF overproduction inhibitor of the present invention is the same. It can be seen that the administration of the product can significantly suppress the overproduction of TNF after the administration of a high dose of LPS.

From the above, it is clear that by the perioperative administration of the inhibitor of the present invention, it is possible to prevent a disorder in which TNF is excessively produced due to surgical invasion or infection, which adversely affects the living body. Became.

[Brief description of drawings]

FIG. 1 is a graph showing changes over time in plasma TNF levels of rats in each group in the test according to Test Example 1.

Claims (2)

[Claims] 1. A TNF overproduction inhibitor comprising inosine, cytidine, uridine, guanosine-5'-1 phosphate or a salt thereof and thymidine as an active ingredient. 2. An excess of TNF according to claim 1, which contains inosine, cytidine, uridine, guanosine-5'-1 phosphate or a salt thereof and thymidine in a molar ratio of 4: 4: 3: 4: 1. Production inhibitor.