CS210659B2 - Method of preparation of amidobenzoic acid derivatives - Google Patents

Abstract

Aminobenzoic acid derivs. of formula (I) and their salts are novel cpds.: (where R1 = H or halo; R2, R3, = H, halo or 1-6C alkyl with the condition that R1, R2 and R3 are not simultaneously H). (I) improve liver function and, strengthen and normalise immunity and antiviral activity and are used to treat immuno-depletion illnesses such as articular rheumatism, auto-depletion illnesses, cancer, bacterial infections and asthma. They are administered in daily doses of 0.5-3000mg (pref. 10-300mg) orally or 0.5-100mg (pref. 1-100mg) parenterally.

Description

The invention relates to novel aminobenzoic acid derivatives, to a process for their preparation and to pharmaceutical preparations containing them.

The aminobenzoic acid derivatives according to the invention can be represented by the general formula I

C1 COOH (I) for improving liver function and as a medicament for the treatment of immune-mediated diseases such as rheumatic arthritis, autoimmune diseases, cancer, bacterial infectious diseases and asthma.

Examples of derivatives falling within the scope of formula I include:

N-2‘-carboxyphenyl-4-chloroanthamic acid,

N-2‘-carboxy-4‘-chlorophenyl-4-chloroanthranilic acid,

N-2‘-carboxy-4‘-methylphenyl-4-chloroanthranilonic acid,

N-2‘-carboxy-4‘-ethylphenyl-4-chloroanthedral acid,

N-2´-carboxy-4´-propylphenyl-4 - ^ (^ H (^ i ^ anthranilic acid,

N-2‘-carboxy-4‘-butylphenyl-4 - [(1 H, 2 H)] - (1 H) anthranilic acid,

^D N-2-carboxy-5'-chlorophenyl-5-chloroanthranilic acid,

N 1 -carboxy-4-methylphenyl-4-chloroanthranilic acid, wherein each of the symbols

R 1 and R ', which are the same or different, represent a hydrogen atom, a halogen atom or an alkyl radical having 1 to 6 carbon atoms,

All aminobenzoic acid derivatives of the invention are novel. The derivatives according to the invention have an excellent effect in improving liver function and strengthening and regenerating immunity and have an antiviral effect.

For these reasons, they are useful as the middle 210659

N-2‘-carboxy-5‘-methylphenyl-4-chloroanthranilic acid,

N-2‘-carboxy-3‘-methylphenyl-4-chloroanthranilic acid,

N-2‘-carboxy-3‘, 6‘-dimethylphenyl-4-chloroanthrylic acid; and

N-2‘-carboxy-5‘-chlorophenyl-4-chloroanthranilic acid.

The present invention relates to a process for the preparation of aminobenzoic acid derivatives of the general formula I, characterized in that the 2-halobenzoic acid derivative of the general formula II

COOH

X

cl (ID where

R1 is as defined above and

X is a halogen atom, reacted with anthranilic acid or a derivative of formula III at a temperature of 80 ° C to 200 ° C

(III) where

Ri and R? are as defined above and in the presence of an alkali and a substance selected from the group consisting of metals, metal and iodine compounds and iodine containing compounds as catalyst.

According to the invention, for example, 2-chlorobenzoic acid, 2,4-dichlorobenzoic acid, 2,4-dibromobenzoic acid and the like can be used as 2-halobenzoic acid derivatives of the formula II according to the invention.

As anthranilic acid derivatives of the general formula III according to the invention, 4-chloroanthranilic acid, 5-chloroanthranilic acid, 4- (4-chloro-3- (4-chloro-3-methyl) -thranilic acid, 3-methylanthranilic acid, 3- methylanthranilic acid, 5-methylanthranilic acid, 3,6-dimethylanthranilic acid and the like.

According to the invention, the reaction of 2-halobenzoic acid derivatives of formula II with anthranilic acid or a derivative thereof of formula III is carried out in a solvent at a temperature above 25 ° C, usually at a temperature of 80 to 200 ° C, for 1 to 10 hours, and preferably at a temperature of 120-150 ° C, for 3-5 hours.

Suitable solvents for the process according to the invention are isoamyl alcohol, dimethylsulfoxide, dimethylformamide, ethanol, butanol, n-amyl alcohol, diethylene glycol dimethyl ether, nitrobenzene, water and the like.

The anthranilic acid or a derivative of formula (III) may advantageously be used in excess, preferably in an amount of 1.2 to 2.0 moles per mole of the 2-halobenzoic acid derivative. Alkali compounds such as potassium carbonate, sodium carbonate or copper carbonate and a metal or a metal containing compound such as powdered copper, copper chloride, copper bromide, copper acetate can advantageously be used to carry out the reaction smoothly and to increase yields of the desired product. and the like as reaction catalyst.

The alkali is preferably used in an amount greater than the total amount of the compounds of formulas II and III in moles, while the metal or metal-containing compound is preferably used in very small amounts. In addition, the product yield can be greatly increased by using a catalytic amount of iodine or an iodine-containing compound, such as sodium iodide or copper iodide, in addition to the catalysts mentioned above.

The aminobenzoic acid derivatives of the formula I can be converted in any conventional manner into the corresponding alkali salts, such as sodium, potassium salts and the like.

Aminobenzoic acid derivatives of general purpose; Formula I can be formulated together with 'excipients and' pharmaceutically 'suitable carrier' pharmaceutical preparations' to 'improve' liver function and 'to pharmaceutical' preparations for the treatment of immune-depleted diseases. These preparations may take a form suitable for oral administration, for example, in the form of tablets, granules, powders, capsules filled with powder or granules and the like, or a form suitable for parenteral administration, such as parenteral injection solutions in water.

Preferably, lactose, starch, dextrin, sucrose, microcrystalline cellulose, kaolin, calcium carbonate, talc, and the like are used as pharmaceutically acceptable carriers for the preparation of tablets, granules, powders or capsules. the solution is isotonized with sodium chloride, potassium chloride and the like.

The aminobenzoic acid derivatives of formula (I) are to be present in the pharmaceutical preparations in an amount which is 'sufficient' to achieve the desired effects, ie.

the effect on improving liver function and the 'immune enhancing effect' and its rege210659 p

not exhausted. The usual dosage is 0.5 to 3000, preferably 10 to 300 mg of active compound per person per day for oral administration, and 0.5 to 1000, preferably 1 to 100 mg of active compound per person per day for administration of parenteral injections.

Fig. 1 is a graph showing the effect of a compound of the invention in the treatment of arthritis with an artificially induced excipient (rheumatic arthritis model).

Fig. 2 is a graph showing the effect of a compound of the invention in the treatment of a model autoimmune disease.

Fig. 3 is a graph depicting the anti-tumor effect of a compound of the invention against transplanted tumor cells (Sarcoma-180).

Fig. 4 is a graph showing the effect of a compound of the invention in promoting lymphocyte blastogenesis [in vitro].

Fig. 5 is a graph showing the effect of a compound of the invention in promoting blastogenesis of lymphocytes (in vivo).

Fig. 6 is a graph showing the effect of a compound of the invention in the regeneration of depleted immunity.

Fig. 7 is a graph showing the effect of a compound of the invention in protecting against Salmonellae infection.

Fig. 8 is a graph showing the effect of a compound of the invention in the treatment of an allergic reaction (IgE formation).

t

In graphs it means - static

Even standard deviation.

The invention is illustrated by the following experiments and examples. These experiments and examples are illustrative only, but do not limit the scope of the invention in any way.

♦

Experiment 1

Effect of decreasing the level of triglycerides in the liver

Female rats (Sprague Dawley) weighing 170 to 200 g without food for 15 hours are divided into groups of 4 rats each. The rats were orally dosed with various test compounds at 100 mg / kg and 30 minutes after the test compound was administered orally with ethanol at 6 g / kg. 24 hours after ethanol administration, the liver of each rat is collected and the level of neutral lipid in the liver is measured in a conventional manner to calculate the% prevention (defined below) against an increase in the level of triglycerides in the liver. One control group is given ethanol, the other control group is given only an aqueous glucose solution with an equivalent calorific value as the same amount of ethanol, otherwise the experiments are carried out in the same way as the test substances. In another positive control group, commercially available 2-mercaptopropionylglycine was used in place of the test compound.

Table 1

Test Compound% Prevention * N-2‘-Carboxyphenyl-4-chloroanthranilic acid67.1

N-2‘-carboxy-4‘-methylphenyl-4-chloroanthranilic acid53,2

N-2‘-carboxy-6‘-methylphenyl-4-chloroanthranilic acid25,2

N-2‘-carboxy-5‘-chlorophenyl-4-chloroanthranilic acid. . 57.2.

N-2‘-carboxy-3‘, 6‘-dimethylphenyl-4-chloroanthranilic acid.45,8 N-2‘-carboxyphenyl-4-chloroanthranilic acid sodium salt63,6 control test:

2-mercaptopropionylglycine 9.3% / prevention is calculated according to the following equation:

amount of triglycerides in the liver group administered ethanol -% prevention = ---------------- amount of triglycerides in the liver group administered ethanol - amount of triglycerides in the liver group test compound which was administered and ethanol

X 100 amounts of triglycerides in the liver group that received glucose

106 5 9

Experiment 2

Effect on bile excretion

a] Bromosulfophthalein (BSP) test

Female rats (Sprague Dawley) weighing 150-190 g without food for 15 hours are divided into groups of 4 rats each. The rats were orally administered various test compounds at a dose of 100 mg / kg and 60 minutes after administration of the test compound, the rats were injected into the tail vein with 28 mg BSP per head, 30 minutes after the rats were injected. with a conventional BSP colorimetric method of concentration. For control, physiological saline was administered orally instead of the test compound.

The results are shown in Table 2.

Table 2

Test compound% BSP balance control 51.9 + 6.7

N-2‘-carboxyphenyl-4-chloroanthotally 17.3 + 8.4 acid ** (P <0.01) ** Significant effect:

Student's T-test based on mean value

(b) Indocyanogree (ICG) test

Female rats (Sprague D.awley) weighing 180-210 g without food for 15 hours are divided into groups of 4 rats each. The rats were orally administered various test compounds at a dose of 100 mg / kg and 60 minutes after administration of the test compound, 2 mg of ICG per head was injected into the tail vein. 15 minutes after injection, the rats are bled from the heart and the ICG concentration is determined by a conventional colorimetric method. As a control, physiological saline was administered orally instead of the test compound.

The results are shown in Table 3.

Table 3

Test compound% ICG balance control (saline) 7.8 + 1.5

N, N -carboxyphenyl-4-chloroanthranilic acid

Female rats (Sprague Dawley) weighing 160-200 g, without food for 15 hours, are divided into groups of 4 rats each. The rats were orally administered various test compounds at a dose of 100 mg / kg and injected 150 g of bilirubin per head into the tail vein 60 minutes after administration of the test compound. Twenty minutes after injection, rats are bled from the heart and the blood concentration of bilirubin is determined in a conventional manner. As a control, physiological saline was administered orally instead of the test compound.

The results are shown in Table 4.

Table 4

Test Compound% bilirubin balance control (physiological saline) 85.4 + 9.8

N-2‘-carboxyphenyl-

-4-chloroanthranilic acid 39.7 + 13.3 acid ** (P <0.01) ** as in Table 2.

Experiment · 3 ^:

Effect in the treatment of arthritis artificially induced by the excipient (rheumatoid arthritis model)

Male rats (Sprague Dawley) 8 weeks old were used in the experiment. Arthritis is induced by subcutaneous injection of a suspension of killed tuberculosis bacilli in Freund's adjuvant into the tail of each rat. 17 days after sensitization, the degree of swelling of the hindpaws of each rat was determined by measuring their volume and the rats were divided into groups of 10 individuals so that the average degree of swelling was approximately the same for each group. Each ski-rat was dosed orally with a test compound which is N-2'-carboxyphenyl-4-chloroanthamic acid disodium salt, at the dose given in Table 5, once daily for 1 week starting on day 17 after sensitization. The degree of hind paw edema is measured every second or third day from administration of the test compound until day 31 after sensitization. Reduction in hindpaw volume is used as a value to evaluate the pharmacological effect. The results are shown in Fig. ·

1. It has been found that administration of the test compound at a dose of 5 or 10 mg / kg produces a significant effect in the treatment of artificially induced arthritis in rats.

The results of the statistical test performed using the values obtained 24 and 31 days after sensitization are shown in Table 5.

as in Table 2

c) Effect on biiliubin eerecia

5.1 + 0.2 ** (P <0.02)

210859

Table 5

Dose (mg / kg) 24 31 day after sensitization

control

0.1

100 ALIGN!

3.693 + 0.130 (ml)

3.524 + 0.170

3,214 + 0,161

3.175 + 0.100 **

3,027 + 0,126 **

3.350 + 0.168

3.646 + 0.170 (ml)

3.762 + 0.216

3.289 + 0.223

3.102 + 0.141 *

3.121 + 0.176 *

3.482 + 0.170 * significant effect: Student's T-test (₽ <0.05) based on mean ** significant effect: Student's T-test (P <0.01) based on mean

Experiment 4

Effect in treatment of model autoimmune disease

The pharmacological effect of the test compound, which is the disodium salt of N-2‘-carboxyphenyl-4-chloroanthranilic acid, is studied in a model autoimmune disease, which is glomerulonephritis of New Zealand female mice (Black / White Fi). The test animals are divided into groups of 8 each and administered orally with the test compound at a dose of 5 or 50 mg / kg per day, starting at 8 weeks of age. The development of the disease is monitored by measuring the urinary protein level using Combistix assay paper (Ames Co., Inc.). Test paper evaluation was found to be in good agreement with the results obtained by quantitative analysis using the sulfosalicylic acid method. The differences in mean urinary protein levels in each group between 20 weeks of age and 35 weeks of age in mice are shown in Figure 2. Figure 2 shows that urine protein levels in control animals increase significantly around time 27 weeks from birth, whereas in all test compound groups, the urine protein level is relatively stable. Thus, the compound of the invention exhibits a pronounced pharmacological effect in suppressing autoimmune; glomeruloneftrltis. In addition, 3 of the 8 mice in the control group sent glomerulonephritis between 35 and 40 weeks after birth, whereas none in the test compound groups received any.

Experiment 5

Antitumor effect in transplanted tumor cells (Sarcoma-180)

Test animals, which are male (ddY) mice weighing 20-25 g, are divided into groups of 9 or 10 individuals. Each mouse is transplanted to the heel of the hind paw by subcutaneous injection of 10 ⁶ tumor cells to induce a solid tumor. The test compound, the disodium salt of the N-2'-carboxyphenyl-4-chlorantranil compound, is administered orally once daily for 11 days from the day of transplantation. 14 days after transplantation, the tumor tissue was harvested from each mouse and its wet weight was determined. This value serves to characterize the antitumor effect of the test compound. The results are apparent from Fig. 3. A dose of 10 mg / kg showed a marked effect on tumor growth suppression. The test compound does not inhibit the growth of the same tumor cells "in vitro" and has no antitumor effect directly on the cells. Therefore, the test compounds are believed to have an effect mediated directly by the host organism.

Experiment 6

Antitumor effect in transplanted cancer cells (AH-13)

8-week-old female rats (Donryu) were used as test animals. The rats are divided into groups of 10 animals each. Each rat is administered intraperitoneally with the disodium salt of N-2'-carboxyphenyl-4-chloroanthranilic acid, as test compound, once daily for a total of 10 days, starting on day 16 before inoculation with tumor cells. 16 days after the first administration of the test compound, 10 ⁶ AH-13 tumor cells are inoculated intraperitoneally to induce abdominal cancer. The mean survival time (days) in rats after tumor cell inoculation is shown in Table 6. It is evident that administration of the test compound at 50 mg / kg achieves an antitumor effect mediated by the host organism.

Table 6

Dose (mg / kg) Survival time (days) Average time

T / K * (%) survival days

200 7 7 7 6 7 6.9 97 7 8 6 7 100 * 7 6 6 7 7 7.3 103 6 6 13 50 7 13 11 15 Dec 11 11.6 163 17 8 10 12 12 control 6 7 7 7 7 7.1 100 ALIGN! 7 6 7 11 6 * Τ / Κ ^f ° / o) __ average time average time survival in survival to the control group X 100

Experiment 7

Effect in promoting blastogenesis of lymphocytes (activation of cellular immunity)

In this experiment, the effect of the test compound, which is the disodium salt of N-2'-carboxyphenyl-4-chloroanthranilic acid, on the blastogenesis of lymphocytes using spleen cells and lymphonoid cells of mice and rats is studied. Male in vitro CsH / He strains are used for the in vitro test and male Wistar-Imammichi in vivo rat tests are used. In the in vitro assay, Concanavallin A, (Sigma) is used as a blastogenetic factor, and the lymphocytes are cultured using RPMI 1640 medium manufactured by Grand Island Biological Co. Cultivation of the lymphocytes was performed in an environment at 37 ° C, 5 ° / o of carbon dioxide for approximately 48 hours then treated with 0.5 μ Ci / ml ³ H-thymidine incorporation, cells were incubated in medium 16 hours and then lymf- the oceans separate.

The radioactivity of the separated cells is measured with a scintillation counter and the value is considered to be an indicator of the degree of blastogenesis. The in vivo test is performed in a similar manner to the in vitro test wherein each rat is dosed orally with a test compound at a dose of 5 or 50 mg / kg. In this case, the rats receiving the test compound are divided into groups of 3 individuals and the cells to be cultured from the rats are maintained in the same culture conditions in each group. The results of the experiments are apparent from Fig. 4 (in vitro) and Fig. 5 (in vivo-).

Giant. 4 clearly shows that the test compound at the concentrations TO '· ¹ to 10 ^_2 g / ml improved activity Concanevallinu A. FIG. 5 shows that the spleen cells and lymphocytes of rats administered the test compound the day before administration of ConcanavaHin A exhibit an SI value of about twice that of control rats, and that ConcanavaHin A activity increases by about 2-fold compared to that of control groups. Groups of rats or mice administered the test compound 4 days prior to the administration or addition of Concanavallin A also show an improvement in activity, but the degree of improvement in activity is lower since the test compound was added or administered earlier.

From the above results, it can be concluded that the test compound improves ConcanavaHin A-based blastogenesis of lymphocytes, not only in vitro but also in vivo, and that it enhances cellular immunity.

Experiment 7

Effect on strengthening Immunological reactions in normal animals (effect against bacterial infection)

Male animals (Sprague Dawley) weighing 200-220 g were used as test animals and divided into 7 groups of 5 or 6 individuals each.

Rats in six groups were orally administered a single dose of the test compound, which is N-2 kar-carboxyphenyl-4-cliloranthranilic acid at a dose of 100 mg / kg. The test compound is administered to each group at six different dates, one day prior to sensitization, the day of sensitization, one day after, two days after, three days after, and four days after sensitization, using the sheep red blood cell ( SRBC), such as an antigen.

SRBC is injected intravenously into the rat through the tail vein in an amount of 1 ml as a 2% aqueous suspension. Control groups were used on the day of sensitization of physiological saline in the same amount and in the same manner as the test groups.

days after sensitization, the rats are sacrificed and the spleen is removed and blood is analyzed.

The number of platelet-forming cells is determined by the Jerne method and the antibody titer is determined by the hemolytic method.

The results are shown in Tables 7 and 8.

Table 7

Increase in the number of platelet-forming cells

Time of administration (day) related to sensitization

Time of administration (day) related to sensitization

Platelet count per 10 ⁶ cells

Platelet count per 106 cells

- 1 93.2 + 28.6 0 193.4 + 26.6 ‘» (P <0.01) + 1 316.8 + 38.2 ** (P <0.01) + 2 76.5 + 16.3 + 3 101.0 + 11.2 + 4 95.5 + 35.3 control 49.7 + 16.0

** very strong effect

Table 8

Antibody titer in blood

Time of administration (day) related to sensitization Time of administration to sensitization (day) related

Final dilution showing hemolysis Final dilution showing hemolysis - 1 + 1 + 2 + 3 + 4 control

29.5

210.8 * (p <o, O5)

213 ** (p <0.01)

29.5

29.8

28.7

29.0 * strong effect ** very strong effect

Table 7 shows that there is a significant increase in the number of platelet-forming cells when rats are administered the test compound on or after the sensitization day. The increment is about six times that of the control group.

Table 8 shows a significant increase in blood titer when rats are dosed with test compound on or after the day of sensitization of SRBC.

Experiment 9 days. Three days . after the first administration, sensitization is carried out with an antigen, which is a sheep red blood cell (SRBC), by injecting the antigen into the tail vein of the mouse. An oral test compound, which is the disodium salt of N-2‘-carboxyphenyl-4-chloroanthranilic acid, is administered to each mouse the day after sensitization. Five days after sensitization, the number of platelet-forming cells (PFC) in sessile is determined in the usual manner. The results are shown in Figure 6.

The number of PFCs in the spleen of mice in the cortisone-treated group was reduced to one third compared to the non-cortisone-treated control group. On the other hand, the number of PFCs in the 10 mg / kg dose of the test compound increases very significantly. The results clearly show that the test compound acts to counteract the immune suppressing effect of cortisone and cause the regeneration of depleted immunity.

The antibody titer in blood determined by the haemolytic method is shown in Table 9.

Table 9

Treatment of Ti'tr antibody * * ^** without treatment 1012 kortison 103 cortisone + test compound (10 mg / kg) 10 ^J 9 koťtison. + test compound (50 · mg / kg) 10 ⁶ * antibody titer in the blood is determined by the degree of dilution at 50% hemolysis. Experiment 10

Effect in improving bactericidal ability in neutrocytes (effect of protecting against bacterial infection)

Male white rabbits of strain JW / CSK weighing 3.0 · 3.9 kg are used as test animals and are divided into groups of five individuals. Each rabbit was orally administered as a test compound with disodium salt of N-2'-carboxyphenyl-4-chloroanthranilic acid at a dose of 100 mg / kg and 24 hours after administration, the neutrocyte in peripheral blood was reacted with nitrobluetetrazorium (NBT) and then treated. determine by microscope the proportion of cells to formazan. The control value is taken from the same animal 3 days prior to administration of the test compound.

The results are shown in Table 10.

The results show that the proportion of formazan-positive neutrocyte cells increases significantly when treated with the test compound,

Effect in the regeneration of depleted immunity (effect in protecting against bacterial infection)

Male mice of IV CS strain of 5 weeks of age are divided into groups of 5 individuals as test animals.

Each mouse is administered intraperitoneally cortisone at a dose of 60 mg / kg once daily for · 5 i

and that the test compound improves the neutrophyte bactericidal ability.

Table 10

NBT treatment * (%) before treatment with test compound 11.4 + 1.8 after treatment with test compound 33.3 ± 7.8 * (P <0.05) * NBT value is reported as "mean + + standard deviation" .

5-week-old male ddY mice, divided into groups of 7 members, are used as test animals. Each mouse is inoculated intraperitoneally with bucillus pyocyaneus, Psendomonas aeruginosa and Pa-П. The test compound, N-2'-carboxyphenyl-4-chloroanthranilic acid disodium, is administered orally once a day - at a dose of 2 mg / head - 6, 5, 4 and 1 days before inoculation and for 54 hours after inoculation the survival time is followed. Inoculation of 2 X 10 5 bacteria per head reveals the activity of the test compound to protect against bacterial infection. Obviously, the test compound reduces the amount of bacillary toxin.

Experiment 11

Efficacy in protecting against infection caused by Bucillus pyocyaneus

Table 11

Number of inoculated bacteria (number per head)

Survival control time number (hours) of dead animals / / number of test animals

Test compound survival time number (hours) of dead animals / / number of test animals

2 X 10® 22 24 24 24 24 7/7 24 24 24 24 24 7/7 26, 26 29, 29 2 X 105 45 45 45 45 48 6/7 46, S, S, S, S 1/7 52, S S, S 2- X 104 52, S, S, S, S, S 1/7 with, with, with, with, with, with 0/7

10.48

1Q5.7

LD50 (number per capita)

S: Live animals 54 hours after inoculation with bacteria.

Experiment 12

Efficacy in protecting against infection caused by Bucillus pyocyaneus

5-week-old male ddY mice were used as test animals and divided into groups of 9 members. 10 mice of Pseudomonas aeruginosa Pa-II are intraperitoneally administered to each mouse and given a single dose of the antibiotic carbenicillin subcutaneously 1.5 hours after administration. The test substance, N-2'-carboxyphenyl-4-chloroanthranilic acid disodium salt, is administered once a day at a dose of 2 mg / head, one and two days before inoculation.

The activity of the test compounds is evaluated by monitoring the test animals for 54 hours of no inoculation. The results are shown in Table 12. The test results show that an improved bactericidal effect is achieved using an antibiotic with the test compound.

Table 12 Dose of carbenicillin Number of surviving animals / number of test animals (mg / head) carbenicillin carbenicillin + tested compound 40 9/9 9/9 20 May 2/9 7/9 10 0/9 4/9 ED05 (mg / head) 24.1 10.4

Experiment 13

Activity to protect against Salmonellae infection

Male animals of the 5 week old ddY strain were used as test animals and divided into 7 groups of 7 members each. Each mouse is inoculated intraperitoneally with 10 ⁵ Salmonella enteritidis TO-1 bacteria and a single dose of 0.5 mg / head is administered subcutaneously 3 hours after inoculation. The test compound, which is the disodium salt of N-2'-carboxyphenyl-4-chloroanthranilic acid, is administered orally at a dose of 2 mg / head on five different days, three days, two days and one day before inoculation, the day of inoculation and a day after inoculation. The results are shown in Table 7. The results show that an improved bactericidal effect is achieved using an antibiotic with the test compound.

Experiment 14

Activity in the treatment of allergic reaction (IgE-producing system) (anti-bronchial asthma effect)

Male mice (Wistar-Imamichi) of 7 weeks of age were used as test animals and divided into groups of 5 individuals. Each rat is sensitized with an antigen, which is an ascaris protein bound to the rest of the dinitrophenol (DNP-AS), with the aid of pertussis vaccine as an adjuvant, which is administered subcutaneously in the back of the animal. The IgE antibody titer is determined 3 hours after intravenous injection of antigen and Evans blue solution inducing an immunological reaction.

The animal is irradiated with 400 X-rays prior to sensitization. The test compound is administered orally once daily for 15 days to 30 days after sensitization. IgE antibody titers determined on 17, 20,

On days 24 and 30 after sensitization, which is determined by a cutaneous response based on a passive cutaneous anaphylactic reaction, they are shown in Figure 8. The results clearly show that the test compound suppresses IgE antibody production and can be used to radically treat allergic diseases.

Experiment 15

Acute toxicity

The acute toxicity of N-2‘-carboxyphenyl-4-chloroanthranilic acid was determined using male and female Wistar-Imamichi rats of 11 weeks of age as test animals. The test compound is administered orally as a suspension in a 5% aqueous gum arabic solution. The results are as follows:

Animal sex LD 50 male 2,100 µg / l <g female 2,600 mg / kg

Example 1

Isoamyl alcohol (250 ml), 2,4-dichlorobenzoic acid (12 grams), anthranilic acid (17.5 g) potassium carbonate · (18 g) are introduced into a 500-ml round-bottomed three-necked flask equipped with a mechanical stirrer and a condenser. ), powdered copper (500 milligrams) and a very small amount of iodine and the mixture was boiled under reflux for 5 hours with stirring. The reaction mixture is allowed to cool to room temperature. Water (500 ml) is added. The mixture is filtered. The filtrate is concentrated under reduced pressure, adding water to remove isoamyl alcohol, then cooled in an ice-water bath and acidified with 6 N hydrochloric acid. The precipitated crystals are separated, suspended in ethanol and the suspension is boiled under reflux for 3-4 hours and then cooled. The precipitated crystals are filtered off, dissolved in tetrahydrofuran, activated carbon is added to the solution and then heated under reflux for 4-5 hours. The activated carbon was separated and tetrahydrofuran was evaporated from the solution under reduced pressure. The resulting residue was resuspended in methanol, boiled under reflux for 3-4 hours and then allowed to cool to room temperature to precipitate crystals. The crystals obtained are filtered to give 14 g of N-2 &apos; -carboxyphenyl-4-chloroanthranilic acid, m.p. &gt; 306 [deg.] C.

For CuH 10 ClNO 2 calculated:

% C, 57.64;% H, 3.45;% N, 80.80;

found:

57.8.1% С, 3.39% Η, 4.50 ^ο / θΝ.

Example 2

The procedure of Example 1 was repeated using 2,4-dichlorobenzoic acid and 5-methylanthranilic acid. There was obtained N-2'-carboxy-4'-methylphenyl-4-chloroanthranilic acid having a melting point of 316-318 ° C (decomposition). Yield 71.5%.

For C 15 H 12 ClNO 4 calculated:

% C, 58.93;% H, 3.95;% N, 4.58; find e n o *

H, 3.89; N, 4.58.

Example 1 a d 3

The procedure of Example 1 was repeated using 2-chlorobenzoic acid and 5-methylanthranilic acid. N-2 &apos; -carboxy-4 &apos; -methylphenylanthranilic acid is obtained having a melting point of 304 DEG C. (yield 72%).

Calcd for C12H18NOt:

H, 4.83; N, 5.16. found:

H, 4.78; N, 5.14.

Example 4

The procedure of Example 1 was repeated using 2,4-dichlorobenzoic acid and 3-methylanthranilic acid. N-2 &apos; -carboxy-6 &apos; -methylphenyl-4-chloroanthranilic acid, m.p. 302 DEG-304 DEG C. (decomposition). Yield = 66%.

For C1SH12CINO4 calculated:

% C, 58.93;% H, 3.95;% N, 4.58; found:

% H, 4.04;% N, 4.59.

Example 1 a d 5

The procedure of Example 1 was repeated using 2,4-dichlorobenzoic acid and 4-chloroanthranilic acid. There was obtained N-2'-carboxy-5'-chlorophenyl-4-chloroanthranilic acid having a melting point above 340 ° C. Yield 59%.

For C 14 H 9 Cl 2 NO 4 calculated:

% C, 51.55;% H, 2.78;% N, 4.29; found:

% C, 51.56;% H, 2.87;

Example 6

The procedure of Example 1 was repeated using 2,4-dichlorobenzoic acid and 3,6 dimethylanthranilic acid. N-2‘-carboxy-3‘, 6‘-dimethylphenyl-4-chloroanthranilic acid is obtained, m.p. 283 ° C. (Yield 45%).

For C 16 H 14 ClNO 4 calculated:

H, 4.41; N, 4.38. found:

H, 4.42; N, 4.39.

Example 1 a d 7

The procedure of Example 1 was repeated using 2-chlorobenzoic acid and 4-chloro-3-methylanthranilic acid. N-2‘-carboxy-5‘-chloro-6‘-methylphenylanthranilic acid, having a melting point of 315 ° C, is obtained. (Yield 65%).

For C 15 H 12 ClNO 4 calculated:

% H, 3.95;% N, 58.53;

1 Я I P7PΠ П ’

% H, 3.97;% N, 4.60.

Example 8

The procedure of Example 1 was repeated using 2,5-dichlorobenzoic acid and anthranilic acid. There was obtained N-2'-carboxyphenyl-5-chloroanthranilic acid having a melting point of 310 ° C. (Yield 60%).

For C 11 H 12 OClNO 1 calculated:

C 57.64, H 3.45, N 4.80; found:

% C, 57.78;% H, 3.52;% N, 4.61.

Example 9

Isoamy lay (50 ml), 2,4-dichlorobenzoic acid (2.4 g), anthranilic acid (3.5 g), anhydrous potassium carbonate (3.6 g) and powdered copper (0.4 g) are introduced. into a 200 ml egg-shaped flask and the mixture is refluxed with stirring with a stirrer for 5 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, water was added to the mixture, and the resulting mixture was filtered with suction.

Water was added to the filtrate, and then the filtrate was freed from isoamyl alcohol by concentration under reduced pressure and acidified with 6 N hydrochloric acid to precipitate crystals. The separated crystals are dissolved in tetrahydrofuran, the solution is treated with activated carbon, then the activated carbon is separated and the solution is concentrated to remove tetrahydrofuran. Methanol was added to the residue, the solution was boiled under reflux and then cooled to room temperature. 1.1 g of N-2‘-carboxyphenyl-4-chloroanthranilic acid are obtained in the form of a yellow amorphous substance. Melting point above 340 ° C. This product is identical to that of Example 1.

Example 10

N-2'-carboxyphenyl-4-chloroanthranilic acid (110 g) was dissolved in a solution of sodium hydroxide (30 g) in water (2 liters) and ethyl alcohol was added. 88 g of the sodium salt of N-2-carboxyphenyl-4-chloroanthranilic acid, mp 388 ^and C (decomposition).

Calcd for CnHeNazCINCl2:

% H, 2.40;% N, 4.17; found:

H, 2.26; N, 4.07.

They did

Pharmaceutical preparations

(a) Tablets. Powdered N-2'-carboxyphenyl-4-chloroalithranilic acid (100 g) was thoroughly mixed with lactose (46 g), microcrystalline cellulose (27 g), corn starch (5 g) and magnesium stearate (2 g), and the resulting mixture was mixed. on a tablet machine it processes into tablets with an average of 8 mm and a weight of 180 mg.

(b) Tablets

The powdered N-2--carboxy-4‘-methylphenyl-4-chloroanthranilic acid (200 g) was sieved through a 50 mesh sieve and then mixed with lactose (173 g) and carboxymethylcellulose calcium (20 g). The mixture is kneaded and the resulting dough is granulated in granulate and dried.

The granules are passed through a 14-mesh sieve and, after addition and mixing with magnesium stearate (3 g), the mixture is formed into tablets having a diameter of 8 mm and a weight of 200 mg.

c) Powder

Lactose (149 g) and magnesium stearate (1 g) are intimately mixed with powdered N-2‘-carboxyphenyl-4-chloroanthranilic acid (250 g) per powder.

(d) Capsules

Lactose (358 g) and magnesium stearate (2g) are added to powdery N-2‘-carboxyphenyl-4-chloroanthranilic acid, mixed thoroughly and filled with hard gelatin capsules. The weight of the capsule is 65 mg, the weight of the powder mixture contained therein is 230 mg.

(e) Granules

The pulverulent N-2‘-carboxy-5 chlor-chlorophenyl-4-chloroanthranilic acid was passed through a 50 mesh sieve and then 200 g of this product was mixed with lactose (173 g) and carboxymethylcellulose calcium (28 g). The mixture is kneaded with a paste made from corn starch (4 g) and water and the resulting dough is granulated in a granulator and dried. The granules are passed through a 14 mesh screen.

f) Suspension

Crystalline cellulose (10 g) and sodium carboxymethylcellulose (0.75 g) were suspended in a solution (400 mL) of 200 g sucrose to form a uniform suspension. Separately, N2‘-carboxyphenyl-4-chloroanthranilic acid (5 g) was pulverized together with sucrose (0.5 g) and water (20 ml) in a ball mill. The originally prepared suspension is added to the powder, then water is added to a total volume of 500 ml. Stir the mixture to homogenise.

g) Parenteral injection

The purified sodium salt of N-2boxy-carboxyphenyl-4-chlorathranilic acid (10 g) and sodium chloride (9 g) were dissolved in distilled water for injection solutions and the total volume was adjusted to 1 liter with distilled water. The solution is filtered with suction through a glass frit (G4) and then filled with 2 ml of transparent injection. The injections are sealed and heat sterilized at 100 ° C for 30 minutes.

Claims (4)

SUBJECT OF THE INVENTION A process for the preparation of aminobenzoic acid derivatives of the general formula I (I) wherein each of the symbols Ri and R? which are the same or different, represents a hydrogen atom, a halogen atom or an alkyl radical of 1 to 6 carbon atoms, characterized in that the 2-halobenzoic acid derivative of the general formula II COOH Cl. (II) where R1 is as defined above-a X represents a halogen atom at 80 to 200 ° C. react with a compound of formula III wherein R1 and R2 are as defined above and in the presence of an alkali and a substance. selected from the group consisting of metals, metal and iodine compounds and iodine containing compounds as catalyst. 2. A process according to claim 1, wherein the reaction is carried out for 1 to 10 hours. 3. A process according to claim 1 wherein the reaction is carried out. at 120 to 150 ° C. pp for 3 to 5 hours. 4. A process according to claim 1 wherein the solvent is a solvent selected from the group consisting of. including isoamyl alcohol, - - dimethyl-;