JPS5980679A - 2-(4(5)-carboxyimidazole-5(4)-carboxamido)-phenylacetamido) carboxymethyl-6h-2,3-dihydrothiazine-carboxylic acid derivative and phagocytic function activating agent containing said compound as active constituent - Google Patents

Abstract

NEW MATERIAL:A compound expressed by the formula (R is hydroxymethyl or 4-sulfoethylpyridinium-methyl). EXAMPLE:2-[ 4(5)-Carboxyimidazole-5(4)-carboxamido-phenylacetamido ]carboxymethyl-5-hydroxymethyl-6H-2,3-dihydrothiazine-carboxylic acid. USE:A phagocytic function activating agent useful for treating infectious diseases and malignant tumors. PROCESS:7-beta-D-(-)-alpha-(4(5)-Carboxyimidazole-5(4)-carboxamido)phe nylacetamido-3- (4-beta-sulfoethylpyridinium)methyl-3-cephem-4-carboxylic acid is hydrolyzed under basic conditions to give the aimed compound expressed by the formula.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides 2-[(4(5)-carboximidazole-
5(4)-Calphox1namide)-Phenylacetamide]
Carboxymedyl-682,3-dihydrothiazine-carboxylic acid derivative (hereinafter referred to as C
STC derivative 1) Contains a phagocytic cell function activator containing the compound as an active ingredient.

Conventionally, chemotherapeutic agents and antibiotics, which suppress the growth of tumors and pathogenic microorganisms, have been mainly used to treat infectious diseases and malignant tumors caused by bacteria and viruses. On the other hand, during these diseases, the functions of the body's phagocytes and lymphocytes (hereinafter referred to as immune function) are generally reduced, so it is possible that the immune function of the body is involved in the overgrowth of these diseases or the progression of the disease. It became clear that there was an important relationship.

Bacteria, viruses, tumor cells, etc. are originally foreign substances to the living body, and are difficult to eliminate by the host's immune system, but if the immune system is weakened, these foreign substances must be sufficiently eliminated. Because it cannot be covered, it is easy to develop the disease, and it is considered to be a cover that can make the disease more severe. Also,
Pathogenic microorganisms and tumor cells are known to suppress immune function, and this is thought to facilitate the progression of infectious diseases and malignant tumors. The treatment of bacterial infections has made great progress with the remarkable development of chemotherapeutic agents, but since many of these drugs suppress immune function, infections caused by attenuated bacteria, which had never been seen before, have increased significantly. ing. Against this background, efforts are being made to treat not only malignant tumors and viral diseases for which there are few effective therapeutic agents, but also bacterial infections by activating the host's immune system. . In order to meet these demands, medicines that activate lymphocyte function have already been developed and are used to treat malignant tumors and infectious diseases, but there are still no known medicines that activate phagocyte function. . As mentioned above, phagocytes are an important factor in the foreign body elimination mechanism, and play a particularly central role in preventing bacterial infection, so there has been a desire to develop a drug that activates this function.

In view of this weight, the present inventors have conducted many years of research to develop a drug that activates phagocytic cell function, and as a result, CST
C derivatives activate various phagocytic functions, preventing eye ulcers and infectious diseases.
J3 J: The present invention was completed based on the discovery that it is effective in treating aggressive malignant tumors.

The CSTC derivative of the present invention can generally be produced as follows. That is, the known compound 7-β-D
-(-)-α-(4(5)-carboximitazole-5(4)-carboxamide)-phenylacetamide 3-(4-β-sulfonidelpyridinium)methyl-
It can be obtained by hydrolyzing 3-cephem-4-carboxylic acid under basic conditions, neutralizing it, and then fractionating and purifying it by column chromatography.

A cs 1-c g conductor made in this way can be pharmacologically converted into a salt form. Examples of the pharmacologically acceptable salt form include alno metal salts such as thorium and potassium, and J:M addition salts with organic bases such as amines.

Both the CS T C derivatives and their salts of the present invention have strong phagocytic function activation, infectious disease treatment and anti-inflammatory effects, and are soluble in water.
It is a compound with excellent properties as a product. Therefore,
The C3TC derivatives and base addition salts thereof of the present invention can be used alone or together with other pharmaceutically active compounds.
Furthermore, if desired, the U Is I1 can be converted into a usable product by adjustment in any of the usual ways (with binders, fillers, fragrances, etc. used in the pharmaceutical industry).

Next, CS T CL'=conductor effectiveness J3 and toxicity will be explained using experimental examples.

Experimental Example 1 Effect on the phagocytic ability of macrophages ICR mice were treated with 2-((4(5)-carboximitazole-5(4)-carboxamide)-
Phenylacetamide] carboxymethyl-5-hydroxymethyl-61-1-2,3-dihydrothiazine-carboxylic acid (C8TC=I) or 2-(
(4(5)-noj) levoxiimidazole-5(4)-
Carboxamide)-Phenylleatamide) Carboxymethyl-5-(4-β-sulfonithylpyridinium)
Methyl-61-1-2,3-dihydrobuazine-carboxylic acid (C8'1-C-If) 10
Injected 7 kg msu intravenously, 24 hours later Hr! Luminal macrophages were isolated. After culturing 1 x 106 cells at 37°C for 20 hours, 2.5 x 1 oll of sheep red blood cells were added and the cells were further cultured for 2 hours. Upon completion of methanol fixation and Giemsa staining, the number of macrophages that had taken up sheep red blood cells was counted, and the ratio of phagocytic macrophages was calculated. The results are not shown in Table 1.

CS-T'C-I and CSTC-If clearly promoted the phagocytic ability of macrophages.

Control 12.8 ± 0.
7C8TC-Ding 22.1 ± 1.1*”★*p
< 0.01 Experimental Example 2 Effect on neutrophil phagocytic ability 1 to peripheral 1f
Neutrophils were collected from i+ in 1!1 minute. The neutrophil 1x105
C3TCI or IC is C8TC-I at 0.1mg/ml
After incubating for 1 hour at 37°C, heat-treated yeast lXl0' was added and further incubated at 37°C for 15 minutes. After methanol fixation and Giemsa staining, neutrophils that had taken up the yeast were counted. and suppressed the proportion of poorly fed neutrophils.

The results are shown in Table 2. J0 cs-rc-r and CS T C-II significantly promoted A-closing of neutrophils.

, 2 table comparison 27.5 ± 1,
0C3T(,-133,,9± 1,5”**p
< 0.01 Experimental Example 3 Effect on the bactericidal ability of neutrophils
bulletin) Volume 24, Page 2175 197
According to 6th grade 1, human neutrophils with polystyrene ladex,
■Nubity (NBT) trial pasting J3J: BiCS -1'
Add C-technique or CS T' C-1 (37℃, 15
After reacting for a minute, 0.5 NICI was added to stop the reaction. Stir at 4°C for 30 minutes, then centrifuge to remove the pellet.
is. Add 1 ml of dimethylformamide to this and bring to 100°C.
1 m of 510 N K 01-1 boiled for 10 minutes at
1 was added, centrifuged, and the absorbance of the upper layer at 710 nm was measured. The results are shown in Table 3.

CSTC-I and C3TC-1t significantly promoted the ability to reduce NB and T, which are indicators of the bactericidal ability of neutrophils.

Control 0.091 ±0. 00
6C8TC-IO, 182±0.007""C3
-IC-1f 0.210±0.009””4*
☆p<0.001 Experimental Example 4 Effect on Pseudomonas aeruginosa infection Group 10 ICR mice were intraperitoneally inoculated with 2.5x 10' of Pseudomonas aeruginosa (IF03445 strain). After 1 hour, CS T C-I or C3TC-II
A dose of 10 ma/ka was intravenously injected, and the survival rate was determined after 12 and 24 hours. The results are shown in Table 4.

CS T' CT and CS T C-II showed a clear protective effect against Pseudomonas aeruginosa infection.

4th ~ Control 50
0C3TC-I 1 70 0 10100 40 C8'1-C-II 1 80 10 10100 50 Experimental Example 5 Acute toxicity experiment Groups of 10 mice and rats each were given solid food and tap water ad libitum. In addition to being fed and reared, C3TC-1 or C3TC-■ aqueous solution was orally administered to examine acute toxicity.

The L Dso value is calculated from the mortality rate for 7 days after one administration,
Litchfield Wilcoxson (1-icMie
It was extracted using the ld-W 1lcoxon) method.

The L D 5o value of CS-C-I and CS-C-n is 5 regardless of whether J3 is in mouse or rat I-! J/kg or more.

As shown in the above experimental examples, C3TC derivatives have the effect of promoting the phagocytosis and bactericidal ability of macrophages and neutrophils, suppress the death of Pseudomonas aeruginosa-infected mice, and have these pharmacological effects. Based on the results of the acute toxicity test, the dose of Expression Test ■ is sufficiently safe. Therefore, C3TC derivatives are extremely useful for the treatment of 8 types of infections and malignant tumors that are closely related to phagocytic cells.

CS T C-I and CS T C-I+ of the present invention are usually used as tJ:04 agents or oral preparations, but they can also be used as inhalation preparations, rectal preparations, or external preparations. The therapeutic dose for adults is 500 to 5000 m (] /day, but it can be adjusted as appropriate depending on the symptoms or route of administration. Oral dosage forms include capsules, tablets, j19. preparations, liquid preparations, injections, etc. For inhalants, for example, lyophilized preparations are prepared by dissolving in distilled water for injection before use; rectal preparations are rectal suppositories; external preparations are liquids and softeners. desirable.
Examples are shown below, but the present invention is not limited thereto.

Example 1 7-β-1,)-(-)-α-(4(5)-carboximidazole-5(4)-carboxinamide)-phenylarethamide 3-(4-β-sulfA edyl pyridinium) knob-ru-3-cephem-4-carboxylic acid 100-9 to 0.1NNa 0+-14,,
The solution was dissolved in Oml and left in a room for 1 hour. 0.5NI
0.5 ml of CI was added and the mixture was compressed under reduced pressure to obtain a 1.5 ml aqueous solution. This solution was applied as a carrier to a column with a diameter of 25 mm and a height of 300 mm.
150ml (solvent methanol) was loaded into a pack and eluted using methanol. The eluate was separated into 101 fractions, the 11th and 12th fractions were combined, and the methanol was distilled off to give slightly yellowish white crystals. 19 more
White crystals were obtained from the second fraction. These two crystals are Ill and C3TC from NMR spectra, respectively.
=I[su1-lium salt and CS T' C-Isu1-
It was determined to be a lithium salt.

C3TC-I storium salt: Melting point 〉300℃ r R (Kf3 r , cmol)i
580.1 390, NMR (DMSO, -1-M5 internal 4F?i*, δ)5
, 1 ・-5, 7 (21-1, 11n,
7, 2-7, 7 (6H, m) CS'IC-II sodium salt: Melting point >300°C 1 F< (KBr, cm') 15ε
30.1390, 1200.105ONMR(1)～+
SO1TMS internal standard, δ)2, 7-2.8 (41
-1, m, 5, 2~5, 7
(2H,m), 7. 3-7.
7 (6t1, nl), 8.0.8.4 (
41,1, dd) Example 2 NoJ Puhir agent CS T C-'I 5000 milk
Sugar 485g Magnesium stearate 15g 000g Weigh the above ingredients and mix them uniformly. Mixed powder to No. 1 hard biratin burel 50011
Fill 1jJ each to make capsules.

Example 3. Tablet CS T C-H 250g Lactose 520 Ribotide starch 150 (+ Polyvinyl alcohol 15g 7-r 7!J Otsu) Add m-soil to τ and 000g After weighing each of the above ingredients, C3TC-I[ Mix lactose J5 and potato starch uniformly. Add an aqueous solution of polyvinyl alcohol to this mixture and prepare wet granules using Method #1. Dry the granules and mix with magnesium stearate. After that, it is compressed into tablets with a weight of 1200 mg.

Example 4. Powder CS l-C-I 100g milk
Sugar 890g - 1 sugar 1>gwj 000g After controlling each of the above ingredients, mix them uniformly to make a 10% defeating agent.

Example 5. Freeze-dried injection CS T C-i Thorium salt 100 (l) was dissolved in 00 ml of distilled water for injection, filtered aseptically using a membrane filter, filled in 10 ml portions into glass containers, and freeze-dried. This is tightly capped to form a lyophilized powder preparation.

Example 6. Freeze-dried inhaler CS T C-IT 100g in distilled water 1000+n
The solution was dissolved in 1 liter of water, filtered aseptically using a membrane filter, filled in 10 ml portions into glass containers, and freeze-dried. Seal this tightly and call it a lyophilized powder preparation.

Applicant: Mochi11 Pharmaceutical Co., Ltd. Ajinomoto Co., Inc. 0 shots Akira Koji Kohji 4-1002-10 Honmoku-cho, Naka-ku, Yokohama 0 shots clear person Yasuo Suzuki 234-29 Sama, Oaza, Kawaguchi City 0 shots Akira Mochida 2-5-4 Komagome, Toshima-ku, Tokyo 0 applicants Ajinomoto Co., Inc. 1-5-8 Kyobashi, Chuo-ku, Tokyo issue Procedural amendment December 1st, 1981 3. Person who corrects Relationship to the incident: Patent applicant Name Mochida Pharmaceutical Co., Ltd. Corrected as ``Rust conductors and their uses''.

Claims (2)

[Claims] (1) 2-[(4(5)-carboxyl Imidazole-5 (4)
-carboxamide)-phenylarethamide J carboxymedyru 6F+-2,3-dihydrothiazine-carboxylic acid M conductor. (2) 2-[(4(5)-) represented by general formula (1)
Carboximidazole-5(4)-carboxamide>
-phenylacetamide] carboxymethyl-6-rubo4-silicic acid derivative.