JPS58219116A - Immunological regulating agent - Google Patents

Abstract

PURPOSE:An immunological regulating agent, containing a trithiazolepentamethinecyanine type compound as an active constituent, administrable continuously for a long term with almost no toxicity nor side effects, effective particularly for chronic rheumarithritis and nephritis, and capable of exhibiting the effect by the oral administration. CONSTITUTION:An immunological regulating agent containing a compound of the formula (R is 1-15C alkyl except n-heptyl; X is halogen, perchloric acid residue, nitric acid residue or organic acid residue) as an active constituent. The compound exhibits activating action in the case of reduced immunological function and suppressing action in the case of sthenia. In the case of normal state, the compound will neither act at all nor exhibit side effects, e.g. cytotoxicity such as reduction in granulocytes, vomitus, nausea, pyrexia, rash or myasthenia. Autoimmune diseases, e.g. chronic rheumarithritis, systemic lupus erythematosus or nephritis, and other immunological insufficiency such as instant and delayed allergic diseases, malignant tumors, serious infectious diseases, etc. may be cited as indications.

Description

DETAILED DESCRIPTION OF THE INVENTION In recent years, various compounds have been developed as immunomodulators, such as levamisole, D-veticillamine, and dinasylium CM-<2-carboxyphenyl)-14-p-roanthranilate. At present, there are only a few products that are completely satisfactory, as they are highly toxic to living organisms.

As a result of intensive research in view of the above circumstances, the present inventors found the general formula (I): (wherein, R is an alkyl group having 1 to 15 carbon atoms (excluding n-heptyl group), The present inventors have discovered that trithiazole pentamethine cyanine compounds represented by atoms, perchlorate residues, nitric acid residues, or organic rR residues are effective as immunomodulators without toxicity or side effects, and have completed the present invention. .

That is, the present invention uses a trithiazole pentamethine cyanine compound represented by the general formula (1) as an active ingredient.
0 11111 Regarding the immunomodulator containing the compound represented by the general formula (1), the compound in which R is an n-heptyl group and X is iodine is a known compound called platonin, and its pharmacological action is antibacterial. It is known to have effects such as promoting wound healing, activating reticuloendothelial hematopoietic organs and endocrine glands, and enhancing antibody production.

However, it has not been previously known that this compound has almost no toxicity or side effects and exhibits excellent immunomodulatory effects, especially when administered orally, and this was discovered for the first time by the present inventor.

In other words, platonin has an activating effect when the immune function is weakened for some reason, a suppressive effect when the immune function is at its peak, and has no effect at all under normal conditions. It has immunomodulatory effects. On the other hand, it does not have the side effects of conventional immunomodulators, such as cytotoxicity such as granulocytopenia, vomiting, nausea, fever, skin rash, and myasthenia, but rather has the effect of maintaining normal cell function. Rather, it is an even more ideal immune intermodal agent, making it possible to administer the drug continuously over a long period of time, and it has been found to be particularly effective against autoimmune diseases such as rheumatoid arthritis and nephritis. .

Furthermore, it has been revealed that it exhibits excellent pharmacological effects, especially when administered orally, at an astonishingly small dose of 1/10 to 1/10,000 of that of other immunomodulators. In addition to the fact that there are almost no
It was found to be a very easy-to-use drug compared to other immunomodulators. Generally, in cases of rheumatoid arthritis, it is difficult to go to the hospital, so oral administration is more preferable than in cases of other diseases.

Therefore, an immunomodulating agent containing pu-nin as an active ingredient will greatly contribute as a preventive and therapeutic agent for immune diseases in general, and particularly as a therapeutic agent for rheumatoid arthritis.

As a result of repeated research with various changes in the alkyl group bonded to the nitrogen atom of the thiazole ring of platonin, the present inventor found that the compound represented by the general formula (■) has the same immunomodulating properties as platonin. They found that it has an effect.

In the above general formula, R is, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, 8.C-butyl, tart-butyl, n-hentyl, isopentyl, 8.0-pentyl, neopentyl, tart-pentyl, n-hexyl, isohexyl,
m@(3-hexyl, neohexyl, tart-hexyl, 5-methylhexyl, octyl, nonyl, decyl,
1 carbon atom excluding n-heptyl such as undecyl, dodecyl, tridecyl, tetradecyl or pentadecyl
~15 alkyl groups, I being chloro, p
The organic acid residues include halogen atoms such as silica, nitrate, perchloric acid residues, nitric acid residues, p-)luenesulfonic acid, nicotinic acid, and orotic acid.

Among these, compounds in which R is a hexyl group or an octyl group and X is iodo are particularly preferred.

The immunomodulator and preferred indications of the present invention include rheumatoid arthritis, systemic lupus erythematosus (EIIIm),
autoimmune diseases, including nephritis, autoimmune hemolytic anemia, and other immediate and delayed allergic diseases;
Other conditions include malignant tumors, immunodeficiency diseases including severe infections, and immunocompromised diseases. Among these, it is particularly effective for chronic rheumatoid arthritis and nephritis.

The immunomodulator of the present invention sufficiently exhibits its effects in a very small amount of 10 to 500 μm per dose for adults (active ingredient equivalent, hereinafter the same). In particular, a dosage of 0 to 100μ2 chives for 1 to 2 days is preferred.

The immunomodulator of the present invention exhibits sufficient activity upon oral administration, and can be used, for example, in the form of tablets, capsules, powder Sf1 granules, liquids, and the like. It can also be used as nasal drops and suppositories. The preparation of the immunomodulator of the present invention is not particularly limited and may be carried out in accordance with conventional methods using commonly used carriers.

Compound (1) used in the present invention can be produced as follows.

2,4-dimethylthiazole (b'p 1415-147%) is synthesized from monocluria heptadone and thioacetamide, and when this is heated and reacted with an alkyl halide at 170°0 in an oil bath, 3-alkyl-2,4 is synthesized. -Dimethylthiazolium halide is obtained. Ethyl orthoformate and acetic anhydride are added to this, the reaction is heated to 140-145°0 in an oil bath, and the obtained crude product is recrystallized from ethanol to obtain the desired 4,4'-dimethyl- 3,3'-Sialyl 8-[2-(4-methyl)-3-alkylthiazole) -2,2'-dicarbosia dihalogenide can be converted.

Table 1 shows the melting points of some of the compounds (I) thus obtained.

Table 1 Next, the immunomodulator of the present invention will be explained with reference to Examples.

Example 1 (Pharmacological test) (1) Adjuvant arthritis suppressive effect 7 to 8 weeks old (7) BD system (around 1509)
Mycobacterium putilicum (+
! ! ycobaaterium butyrium
) suspended in liquid paraffin at a concentration of 12 my/ml was injected into the right hind paw at a rate of 0.05 per rat to induce adjuvant arthritis (AA). The volume of both hind paws was measured by the mercury exclusion method until 50 days after the adjuvant injection, and AA was evaluated.

(a) Preventive effect Compounds A1 and 4 and Pra) were used as test compounds to examine their AA preventive effects.
Start the main administration at the same time as the adjuvant administration, and
It was administered orally for 60 days. The results are shown in Figure 1. FIG. 2 shows the results of administering D-penicillamine and levamisole in place of the test compound in the same manner.

As a result, compound (1) has a 0.05
It was found that muA was suppressed at a dose as low as μ9/mouse, but D-penicillamine and levamisole did not suppress AA even at a dose of 1.5 mg/mouse.

(b) Using the therapeutically effective compounds AI, A4 and platonin as test compounds, their AA? In order to have a lateral effect on the therapeutic effect, the main administration was started at 14 sips after administration of the adjuvant and was orally administered once a day for 40 days. The results are shown in Figure 6.

As a result, compound (1) suppressed muA at a low dose of 0.05 μg/mouse, which is similar to that of pranin, but 1 μg/mouse suppressed muA.
No inhibitory effect was observed at doses of compound (I) of four or more;
It was found that there is an optimal dosage.

(2) Body weight change During the test period for the preventive effect of the adjuvant in (1) (a) above, the arthritis suppressive effect
The body weight of each rat on day 1 and 60 days was measured. The result numbers are shown in Figures 4 and 5. The results showed that no difference was observed in the body weight changes of rats when D-penicillamine and levamisole were administered compared to the control rats, but compound (1)
．． 05μg/l! ! l! Despite the decrease in swelling of the paws, as with pranin, a clear increase in body weight change was observed in the rats compared to the control, and a marked improvement in the overall condition was also observed.

(3) Effect B on PFO response using mouse tile cells
ELlb/l', anti-5RBO using ``rus tile cells''
Compounds A1-4 and planin were used for the PFO reaction.

BRBO 1×10′ lung cells of 13ajb/cv mice
and RPM containing 10% tallow serum with test compound.
00 incubator (57°0
) for 4 days using the Maf'brook method, and the number of direct Pros that appear was calculated using Terne's method (8
cienoe s vol. 140, p. 405, 1965)
It was measured with The results are shown in Figure #I6.

As is clear from Figure M6, the anti-5R of ~1 human compound (I)
BOP? As for the effect on o-response, a biphasic effect was observed, similar to that of Plato and O-1, with an activation effect at low concentrations and an inhibitory effect at high concentrations.

(4) Effects on the rejuvenation reaction using mouse tile cells
mitogen (mitogen) using zbA mouse female cells.
The effect of the test compound used in (8) above on the lymphocyte rejuvenation response induced by LPS (n: LPS) stimulation was investigated.

RPM containing 10% tallow serum with 1X Bajb/c mouse tile cells and test compound
One microculture plate was cultured in Ni 1640 medium in a 002 incubator for 48 hours.

Then %o, 5poi of mono-thymidine (3H-T, I
After further culturing for 18 hours, the cells were collected using a cell harvester (Labo Matshu), and the radioactivity of "a-?(IH) taken into the cells was measured. Electrostatic formation is shown in Figure 7. ., as is clear from the 7th W.J.
11f! Mitogen LPS 20μF1/rnl
When stimulated by pranin, Compound (I) had a significant inhibitory effect similar to that of planin.

(5) Acute toxicity +1 CIY male mouse (5-6 weeks old) (body weight 25-5
09) was used in 8 animals per group, and pranin suspended in 5% gum arabic solution and compounds (1) of Compounds A1 to 4 were administered intraperitoneally, intravenously, and orally with a gastric tube.
Observations were made for 7 days, the number of deaths was determined, and the LD5o value was determined by the van der Wuerden method. The results are shown in Table 2.

Table 2 As shown in Table 2, the LDao value in the case of intraperitoneal administration is 25-4 to 500 my1 kg body weight, and the LD5o value in the case of oral administration is 1.491 kg body weight or more. ) is effective in suppressing arthritis at 0.05μ/mouse,
It can be seen that the immunomodulator of the present invention has an extremely wide safety margin.

The following various dosage forms of immunomodulators were prepared using Compounds A1 to A4 as active ingredients, respectively.

(1) Tablets Tablets with the following formulation were prepared by a conventional method.

Ingredients my/@active ingredient
0.05 Lactose 79.95 Cornstarch 62.50 Sucrose lI
Improper ester 7.50 Total 150 In the case of a gastric soluble agent, it was coated with 5fi% of Tc-5 and then coated with sugar. For enteric coated agents, HP-5
After applying a 10 weight N% coating of No. 5, sugar coating was applied.

(2) Capsules Capsules with the following formulation were prepared using a conventional method. Ingredients mgl gas/active ingredients
0.05 Lactose 146.95 Days W Narrow Ester 5.00 Total 150 (8) Powder A powder with the following formulation was prepared by a conventional method.

Town Active ingredient 0.05 Milk Sugar Sugar 499.95 Total 500 (4) Suppositories The following two types of suppositories were prepared by conventional methods.

Suppositories A Ingredients mp/drug active ingredients
0.05 PEG + 1000 1440 PEG
4000 59.95 Total 1500 Suppository B Ingredients 4 active ingredients
0.05 Uitepzoil H-151280 Uitepsol H-85519.95 Total 1600 (5) Siltubu Siltubu of the following prescription (one dose: 50p915ml was prepared by a conventional method).

Ingredient Content 〇〇 mj Active ingredient 1 ■ Sugar 60g Glycerin
10g Sodium citrate 0,
1) Sodium benzoate 0.
6g saccharin sodium
0.1g purified water appropriate amount

[Brief explanation of the drawing]

Fig. 1 is a graph showing the preventive effects of test compounds on adjuvant-mediated arthritis in rats; Fig. 2 is a graph showing the preventive effects of D-nidiramine and levamisole on adjuvant arthritis in rats; Fig. 3 is a graph showing the preventive effects of test compounds on adjuvant arthritis in rats; Figure 4 is a graph showing the therapeutic effect of the test compound on arthritis. Figure 4 is a graph showing the effect of the test compound on the body weight of rats with adjuvant arthritis.
Figure 6 is a graph showing the effects of D-penicillamine and levamisole on the body weight of rats with adjuvant arthritis.
The figure is a graph showing the effect of the test compound on the pya response using mouse tile cells, and FIG. 7 is a graph showing the effect of the test compound on the rejuvenation reaction using mouse tile cells. target@enemy e wo Matata Hemorrhoid (×) Ichiyokokan C Fude Kimpu (X) 4φki C personnel 稗 (×) Fu −N (cI−) −! 4 ya (ka) 26 countries/Platonin. Methyl test sample A and amount of compound Q-Regular procedure amendment (voluntary) July 9, 1980 Director of the Patent Office Kazuo Wakasugi 1 Indication of case 1989 Patent Application No. 102595 2 Name of invention Immunomodulator 3.Relationship with the case of the person making the amendment Patent applicant address: 4, Hanajiri Kikiyo-cho, Okayama City, Okayama Prefecture Agent address: 540 5. Subject of amendment (1) Drawing 6wI Correct contents (1) Drawing (#j5 drawing) shall be amended as shown in the attached amended drawing (Figure 5). 7 List of attached documents (1) Amended drawings (Fig. 5) 1 written amendment (toge) August 7, 1980 Commissioner of the Patent Office Kazuo Wakasugi 1 Indication of case 1988 Patent Application No. 102393 2. Name of the invention: Immunomodulator 3. Relationship with the person making the amendment Patent applicant 4. Attorney
540 Subject of 5 amendments (1) Drawing Contents of 6 amendments (1) Drawing (Fig. 3) attached as an attached amended drawing (Fig. 3)
Correct as shown in the figure). 7 List of attached documents

Claims (1)

[Claims] 1 General formula (1): (wherein R is an alkyl group having 1 to 15 carbon atoms (excluding niheptyl group), x is a hapgen atom, a perchloric acid residue, An immunomodulator containing a tri-thiazole pentamethine cyanine compound represented by nitric acid residue or organic acid residue as an active ingredient. 2. The immunomodulator according to claim 1, wherein X in the general formula (1) is iodine. 3. The immunomodulator according to claim 1 or 2, wherein R in the general formula (1) is an octyl group. 4. The immunomodulator according to claim 1, 2, or 3, wherein the indication is rheumatoid arthritis. 5. The immunomodulator according to claim 1, 2, 3, or 4, wherein the administration form is an oral dosage form.