Abstract:
Disclosed is a pharmaceutical composition containing an aminobenzoic acid derivative as an active ingredient represented by the following general formula: ##STR1## wherein  1  R denotes one member selected from the group consisting of the residual groups formed by removing OH at 1(alpha) or 1(beta) position from arabinose, xylose, glucose, galactose, rhamnose and mannose, and  2  R denotes hydrogen or methyl group, or a pharmaceutically acceptable salt thereof.

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This application is a divisional of application Ser. No. 84,467 filed, Oct. 12, 1981, now U.S. Pat. No. 4,322,409 which in turn is a continuation-in-part of Ser. No. 24,092, filed Mar. 26, 1979, now abandoned. 
    
    
     BACKGROUND OF THE INVENTION 
     The present invention relates to a phermaceutical composition containing an aminobenzoic acid derivative as an active ingredient represented by the following general formula: ##STR2## wherein  1  R denotes one member selected from the group consisting of the residual groups formed by removing OH at 1 (alpha) or 1 (beta) position from arabinose, xylose, glucose, galactose, rhamnose and mannose, and  2  R denotes hydrogen or metyl group, or a phermaceutically acceptable salt thereof. 
     The inventors of the present invention, during the course of searching chemical compounds having antitumor activity, have found that chemical compounds represented by the above-mentioned formula (1) have respectively a number of physiological activities such as blood sugar-reducing activity, antihypertensive activity, blood lipid-reducing activity, antiinflammatory activity and central nerve-depressing activity in addition to its antitumor activity. 
     Although the above-mentioned aminobenzoic acid derivatives are known compounds, no report has been found on the physiological activity of the compounds. 
     &#34;Inoue, et al. N-Glycosides. XIX. Preparation of anthranilic acid N-glycosides., Chemical Abstracts, Vol. 48 (1954), Column 2001 i.&#34; and &#34;Inoue, et al. N-Glycosides. XXV. Paper chromatography of N-glycosides., Chemical Abstracts, Vol. 48 (1954), Column 2003 a.&#34; disclose the chemical syntheses of the compounds which are the active ingredients of the phermaceutical composition of the present invention. However, there is no utility disclosed in this prior art and no teaching of pharmaceutical &#34;dosage unit forms&#34;. 
     Furthermore, although U.S. Pat. No. 2,659,689 discloses a p-aldimino benzoic ester and a composition for protecting the human skin from erythema producing rays, the composition comprising a solution of p-aldimino benzoic ester, there is no teaching of pharmaceutical &#34;dosage unit forms&#34;. 
     Accordingly, the present invention has the object of providing a pharmaceutical composition having effectiveness in antitumor activity, blood sugar-reducing activity, antihypertensive activity, blood lipid-reducing activity, antiinflammatory activity and central nerve-depressing activity, based on the discovery of the new medical use of the chemical compounds represented by the above-mentioned formula (1). 
     The present invention will be explained in detail in the following. 
    
    
     BRIEF DESCRIPTION OF THE DRAWING 
     The annexed FIGS. 1 to 13 show respectively the infrared absorption spectra of respective compound No.1 to No.13 in Table 1. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The active ingredient of the pharmaceutical composition of the present invention is a compound represented by the following formula: ##STR3##wherein  1  R and  2  R are as described above, or its pharmaceutically acceptable salt selected from the group consisting of itsNa-, K-, 1/2Mg-, 1/2Ca- and 1/3Al-salts. The sugar moiety of the active ingredient has a structure of six membered heterocyclic ring. 
     The method of preparation of the active ingredient of the present inventionis illustrated as follows: 
     A mixture of 4.5 to 5 g of aminobenzoic acid, 5-6 g of mono saccharide (L-arabinose, D-xylose, D-glucose, D-galactose, L-rhamnose or D-mannose) and 0.1 to 0.5 g of ammonium chloride (formic acid, hydrochloric acid, acetic acid, or magnesium chloride) was heated in 40 to 90 ml of 95 to 100% ethanol or pure methanol under a reflux condenser to induce condensation. After the reaction is over, the reactant is left at room temperature or in a cool place and the crystals separated out are collected by filtering the reactant solution. These crystals are washed with water, ethanol or ethyl ether, and then recrystallized from methanol,ethanol or an aqueous solution of methanol or ethanol. 
     In order to substitute the hydrogen atom of the carboxyl group of the thus prepared compound with a base, it is preferable to follow the known method. The compound, ortho-aminobenzoic acid --N-pyranoside, is dissolvedin an aqueous ethanolic solution and an inorganic salt is added to the solution to effect the substitution. 
     The physical properties of the compounds (the active ingredient of the pharmaceutical composition of the present invention) prepared by the above-mentioned methods are shown in Table 1, and their infrared absorption spectra are respectively shown in FIGS. 1 to 13. Methods of determination of the physical properties are as follows. 
     
                                           TABLE 1__________________________________________________________________________Physical Properties of the Active Ingredients                           Specific                                  Elementary                                          Ultraviolet                  Melting point                           rotation                                  analysis (%)                                          absorption MaximumCompound               (°C.)                           |α|.sub.D.sup.20                                  C:H:N   (millimicron)__________________________________________________________________________  o-Aminobenzoic acid-N--L-arabinoside                  167      -11 (10° C.)                                  50.1:6.0:5.1                                          330, 250, 220                           in ethanol                                   (50.2:6.0:4.9)*  Sodium o-aminobenzoate-N--L-arabinoside                  155-162(decomp.)                           -44    46.6:5.1:4.6                                          315, 246, 212                           in water                                  (46.6:5.2:4.5)  o-Aminobenzoic acid-N--D-xyloside                  168(decomp.)                           +11 (18° C.)                                  53.4:5.7:5.0                                          330, 250, 220                           in ethanol                                  (53.5:5.6:5.2)  Sodium o-aminobenzoate-N--D-xyloside                  160-170(decomp.)                           +1     49.7:4.8:4.7                                          316, 248, 215                           in water                                  (49.5:4.8:4.8)  o-Aminobenzoic acid-N--D-glucoside                  137-138(decomp.)                           +68 (15° C.)                                  49.0:6.1:4.2                                          330, 250, 220                           in ethanol                                  (49.2:6.0:4.4)  Sodium o-aminobenzoate-N--D-glucoside                  145-160  +6     46.1:5.2:4.0                                          318, 249, 215                           in water                                  (46.0:5.3:4.1)  o-Aminobenzoic acid-N--D-galactoside                  152      -16    52.3:6.0:4.8                                          330, 250, 220                           in ethanol                                  (52.2:5.7:4.7)  Sodium o-aminobenzoate-N--D-galactoside                  157-163(decomp.)                           -9     48.5:5.2:4.4                                          317, 248, 215                           in water                                  (48.6:5.0:4.4)  o-Aminobenzoic  acid-N--L-rhamnoside                  165-166(decomp.)                           +52    54.8:5.9:4.9                                          330, 250, 219                           in ethanol                                  (55.1:6.0:4.9)10.  Sodium o-aminobenzoate-N--L-rhamnoside                  152-162(decomp.)                           +54    51.0:5.4:4.9                                          320, 249, 215                           in water                                  (51.1:5.2:4.6)  o-Aminobenzoic acid-N--D-mannoside                  150-165(decomp.)                           -10    51.9:5.8:4.7                                          333, 249, 218                           in ethanol                                  (52.2:5.7:4.7)  Sodium o-aminobenzoate-N--D-mannoside                  148-167(decomp.)                           -2     46.1:5.5:4.0                                          330, 250, 219                           in water                                  (46.0:5.3:4.1)  Methyl o-aminobenzoate-N--D-mannoside                  177-178  -54    49.1:6.1:4.3                                          330, 251,                           in ethanol                                  (48.1:6.6:4.0)__________________________________________________________________________Note:(: : ): theoretical values of C, H and N (%). 
    
     (1) Melting point: determined by the use of micro melting point determination apparatus made by Yanagimoto Works, Japan. 
     (2) Specific rotation: determined by using direct-reading polarimeter ModelOR-50 made by Yanagimoto Works, Japan, with a thickness of 50 mm of an aqueous ethanolic solution of the acidic active ingredient and an aqueous solution of the sodium salt of the acidic active ingredient. 
     (3) Molecular composition: Elementary analysis was carried out by using CHN-Coder Model MT-2 made by Yanagimoto Works, Japan. 
     (4) Ultraviolet absorption spectrum: by using self-recording spectrophotometer Model PS-3T made by Hitachi Works, Japan, on an aqueous ethanolic solution of the acidic active ingredient and on an aqueous solution of the sodium salt of the acidic active ingredient of the medicine. 
     (5) Infrared absorption spectrum: determined by KBr-method using infrated absorption spectrometer Model DS-701G made by Nippon Bunko Co. Ltd. Japan.The chart number of the spectrogram coincides with the number of specimens of the active ingredient of the pharmaceutical composition. 
     The following are the physiological properties of the active ingredient of pharmaceutical composition of the present invention described in the orderof (1) acute toxicity, (2) anti microbial activity, (3) mutagenicity, (4) delayed-type intracutaneous reaction and (5) antibody-producing activity. 
     (1) Acute toxicity 
     Acute toxicity of the active ingredient was examined by respective intraperitoneal and oral (forcible) administration to ICR-JCL mice. The specimen was dissolved in the physiological saline solution in intraperitoneal administration, and dissolved in distilled water in oral administration. 
     Their symptoms were observed after administration until the 7th day of administration, and LD 50  of the specimen was obtained from the mortality accumlated to the 7th day, according to the graphic method of Litchfield-Wilcoxon. The results are shown in Table 2. As is seen in Table2, more than half of the active ingredients are qualified to be highly safeactive ingredients of the medicine. 
     
                       TABLE 2______________________________________Acute toxicity of the active ingredients(LD.sub.50 in g/kg)                Route of                administration                  intraperi-Compound               toneal   oral______________________________________Sodium o-aminobenzoate-N--L-arabinoside                  7.48     6.35Sodium o-aminobenzoate-N--D-xyloside                  9.27     6.35Sodium o-aminobenzoate-N--D-glucoside                  13.38    8.96Sodium o-aminobenzoate-N--D-galactoside                  7.42     6.10Sodium o-aminobenzoate-N--L-rhamnoside                  &gt;15.00   12.50Sodium o-aminobenzoate-N--D-mannoside                  &gt;10.00   &gt;10.00Methyl o-aminobenzoate-N--D-mannoside                  2.5      &gt;7.50______________________________________ 
    
     (2) Anti microbial activity 
     The active ingredient was dissolved in distilled water at a series of two fold dilution system. These diluted solutions were mixed with agar medium in 9 times by volume and the mixture was poured into a petridish. Heartinfusion agar medium was used for bacteria, and Sabouraud&#39;s agar medium was used for fungi. After streaking with the pre-culture, the inoculated plates were incubated at 37° C. for 20 to 24 hours for bacteria and at 25° C. for 3 to 7 days for fungi, and then the growth was examined. The following microorganisms were used for assessing the antimicrobial activity: 
     Pseudomonas aeruginosa IAM 1514 
     Escherchia coli IFO 12734 
     Staphylococcus aureus 209 P 
     Bacillus subtilis IAM 1069 
     Saccharomyces cerevisiae IAM 4207 
     Candida albicans ATCC 752 
     Trichophyton mentagrophytes IFO 6124 
     Aspergillus niger IAM 3001 
     As the result of the above-mentioned tests, it was found that none of the tested active ingredients showed growth inhibition of the microorganisms at a concentration of 1 mg/ml. 
     (3) Mutagenicity 
     As the first stage, the active ingredients were tested by rec-assay (i), and as the second stage, they were tested by reversion assay (ii). 
     (i) A strain of Bacillus subtilis M 45, a defectant of recombination repair, and a wild strain of Bacillus subtilis H 17 keeping recombination repair activity were inoculated to make their own streaks not crossed at the start on a B-2 agar culture plate (made by dissolving 10 g of meat extract, 10 g of polypeptone, 5 g of sodium chloride and 15 g of agar in 1000 ml of distilled water at a pH of 7.0). Then, a circular sheet of filter paper 8 mm in diameter, which absorbed 0.04 ml of an aqueous solution of the active ingredient (using sterilized water) was put on the surface of the agar plate so as to cover the starting point of the above-mentioned streaks of bacterial culture. The inoculated B-2 agar culture was kept at 37° C. for a night and the length of growth-inhibited region was measured. Kanamycin was used as the negative control and Mitomycin C was used as the positive control. The results of the rec-assay are shown in Table 3. 
     (ii) The strains TA 98 and TA 100 (both are histidine requiring) of Salmonella typhimurium were used in the reversion assay. 
     Into 2 ml of a soft agar culture medium (the medium itself contains 6 g of sodium chloride and 6 g of agar in 1000 ml of distilled water) to which one tenth by volume of an aqueous solution of 0.5 mM of biotin and 0.5 mM of histidine had been added, 0.1 ml of the bacterial suspension and 0.1 mlof an aqueous solution of the active ingredient were admixed and the mixture was layered on the minimum agar culture medium. After 2 days of incubation at 37° C., the number of revertant colonies was counted.Furylfuramide (AF-2) was used as the positive control. The results of the reversion assay are shown in Table 4. 
     As is seen in Table 3, the active ingredients showed a weak mutagenicity only at a high concentration of 5000 microgram/disk. And as is seen in Table 4, the rate of occurrence of mutation by the active ingredient of the pharmaceutical composition of the present invention did not shown any difference from that in the control to which no substance was added, even at a high concentration of 5000 microgram/plate. These findings demonstrate that the active ingredient is safe from the standpoint of mutagenicity. 
     
                       TABLE 3______________________________________Result of rec-assay             Length of growth-             inhibition zone          Concen-                *differ-          tration  M 45    H 17  enceCompound       (μg/disk)                   (mm)    (mm)  (mm)______________________________________Sodium o-aminobenzoate-          500      0       0     0N--L-arabinoside          5,000    8       4     4Sodium o-aminobenzoate-          500      0       0     0N--D-xyloside  5,000    7       3     4Sodium o-aminobenzoate-          500      0       0     0N--D-glucoside 5,000    5       2     3Sodium o-aminobenzoate-          500      0       0     0N--D-galactoside          5,000    6       1     5Sodium o-aminobenzoate-          500      0       0     0N--L-rhamnoside          5,000    6       2     4Sodium o-aminobenzoate-          500      0       0     0N--D-mannoside 5,000    7       1     6Methyl o-aminobenzoate-          500      0       0     0N--D-mannoside 5,000    7       3     4Kanamycin      10       5       4     1Mitomycin C    0.05     12      2     10______________________________________Note:*difference = length of inhibition zone of M 45 minus length of inhibition zone of H 17. 
    
     
                       TABLE 4______________________________________Results of Reversion assay                  Number of rever-          Concen- tant colonies            tration   (n/plate)Compound         (μg/plate)                      TA 100    TA 98______________________________________Sodium o-aminobenzoate-N--            5,000      59       4L-arabinosideSodium o-aminobenzoate-N--            5,000     166       4D-xylosideSodium o-aminobenzoate-N--            5,000     151       5D-glucosideSodium o-aminobenzoate-N--            5,000     151       6D-galactosideSodium o-aminobenzoate-N--            5,000      61       9L-rhamnosideSodium o-aminobenzoate-N--            5,000      91       7D-mannosideMethyl o-aminobenzoate-N--            5,000      95       8D-mannosideFurylfuramide    0.1       911       167Control (nothing added)            --        149       13______________________________________ 
    
     (4) Delayed-type intracutaneous reaction 
     In order to know the effects of the active ingredients on cellular immunity, the foot pad reaction test was carried out using ICR-JCL mice asexperimental animals and erythrocytes of sheep as an antigen. 
     A mouse was primary-sensitized by injecting 0.2 ml of 10% suspension of sheep erythrocytes in physiological saline solution from the caudal vein and after 7 days of the first sensitization, 0.05 ml of 40% suspension of sheep erythrocytes in physiological saline solution was injected in the foot pad for the second sensitization. The thickness of the foot pad was determined on the next day. The administration of the active ingredient ofthe pharmaceutical composition of the present invention was carried out at the dosage of 250 mg/kg/day once a day for consecutive 5 days centering around the day when the first sensitization was carried out. 
     As the result, the increment of the thickness of the food pad of the mouse administered with the active ingredient showed no significant difference as compared to the increment in the group of mouse not administered with the active ingredient. 
     (5) Antibody-producing activity 
     In order to know the effects of the active ingredients on humoral immunity,the hemagglutination test was carried out using ICR-JCL mice sensitized with sheep erythrocytes. 
     A mouse was sensitized by injecting 0.2 ml of 10% suspension of sheep erythrocytes in physiological saline solution from the caudal vein and after 7 days of sensitization the mouse blood was sampled for the hemagglutination test of determination of the antibody-producing activity.The active ingredient was administered for consecutive 5 days centering around the day of sensitization, intraperitoneally at the dosage of 250 mg/kg/day. 
     As the result, there was no significant difference in agglutination titer between the group administered with the active ingredient and the control group. 
     The following are the pharmacological properties of the active ingredients of the pharmaceutical composition of the present invention described in the order of (1) blood sugar-reducing activity, (2) antihypertensive activity, (3) antitumour activity, (4) analgetic activity, (5) antipyreticactivity, (6) antiinflammatory activity, and (7) blood lipid reducing activity. 
     (1) BLOOD SUGAR REDUCING ACTIVITY 
     Streptozotocin was administered intraperitoneally to a group of Wistar ratsat a dosage of 60 mg/kg and after confirming the positivity of urinary sugar of the animals on the 8th day, regular insulin was further administered to the rats to reduce both the uninary sugar and the blood sugar. Out of thus treated animals, those which certainly showed a higher urinary sugar value and also a higher blood sugar value after a few days of insulin-administration were used as the model animals suffering from artificial diabetes mellitus. The active ingredient was administered to the model animals orally as a solution in distilled water at the respective dosages of 30 and 300 mg/kg. Blood specimens were collected after 3 and 6 hours of the administration, and the determination of glucose in the specimen was carried out by using a RaBA-kit (made by Chugai Pharmaceutical Co., Japan) according to the enzyme method. 
     The results are shown in Table 5. As is seen in Table 5, the difference between the values of blood sugar before and after the administration of every active ingredient (Δ value) was larger than the Δ value of control. 
     Especially, the activity of reducing blood sugar was conspicuous in the case of sodium o-aminobenzoate-N-L-arabinoside, sodium o-aminobenzoate-N-D-glucoside, sodium o-aminobenzoate-N-L-rhamnoside, and sodium o-aminobenzoate-N-D-mannoside, the Δ value of these compoundsbeing 150 to 460 mg/dl at the dosage of as low as 30 mg/kg. 
     
                       TABLE 5______________________________________Blood sugar-reducing activity                    Change .increment. value                   mg/dl of blood            Dose   sugar afterCompound           (mg/kg)  3 hr     6 hr______________________________________Sodium o-aminobenzoate-N--L-              30       -254     -225arabinoside        300      -108     -83Sodium o-aminobenzoate-N--D-              30       -62      -60xyloside           300      -80      -76Sodium o-aminobenzoate-N--D-              30       -220     -200glucoside          300      -134     -217Sodium o-aminobenzoate-N--D-              30       -82      -42galactoside        300      -95      -49Sodium o-aminobenzoate-N--L-              30       -156     -124rhamnoside         300      -462     -397Sodium o-aminobenzoate-N--D-              30       -180     -110mannoside          300      -230     -150Methyl o-aminobenzoate-N--D-              30       -121     -86mannoside          300      -166     -147Control            --       -36      -39______________________________________ 
    
     (2) ANTIHYPERTENSIVE ACTIVITY 
     An aqueous solution of the active ingredient in distilled water was orally administered to rats of spontaneous hypertension at respective dosages of 30 and 300 mg/kg and their blood pressure was determined after 3 and 6 hours of administration by a sphygmomanometer (made by Ueda Works, Japan, Model USM-105R). The difference of blood pressures before and after the administration was used to evaluate the antihypertensive activity of the active ingredients. Mean value of blood pressure of the abovementioned rats in spontaneous hypertension was 200 mmHg. 
     The results are shown in Table 6. As is seen in Table 6, all the tested active ingredients clearly showed that antihypertensive effect. 
     
                       TABLE 6______________________________________Antihypertensive activity                  Reduced amount of                  blood pressure           Dose   after            rate  3 hr     6 hrCompound          (mg/kg)  (mm Hg)______________________________________Sodium o-aminobenzoate-N--L-             30       13        9arabinoside       300      22       25Sodium o-aminobenzoate-N--D-             30       16       16xyloside          300       9        5Sodium o-aminobenzoate-N--D-             30        8        5glucoside         300      26       20Sodium o-aminobenzoate-N--D-             30        9       16galactoside       300       9        8Sodium o-aminobenzoate-N--L-             30       13       19rhamnoside        300      28       20Sodium o-aminobenzoate-N--D-             30       12       14mannoside         300      24       22Methyl o-aminobenzoate-N--D-             30       18       14mannoside         300      26       21Control           --       -2*       2______________________________________Note:-*Blood pressure raised by 2 mm Hg. 
    
     (3) ANTITUMOUR ACTIVITY 
     Sarcoma 180 were transplanted subcutaneously into the right axillary of ICR-JCL mice at the rate of 1×10 6  cells/mouse, and from about 24 hours of transplantation an aqueous solution of the active ingredient is sterilized physiological saline solution was orally administered every other day at a dose rate of 500 mg/kg, 10 times in all. On the 25th day ofthe transplantation, the nodular tumour(s) was extirpated and weighed. 
     The inhibition ratio (I.R.) (%) of the active ingredient was calculated by the following formula: 
     
         (1-T/C)×100=I.R. (%) 
    
     wherein 
     T: mean weight of the tumour(s) in treated group of mice 
     C: mean weight of the tumour(s) in control group* of mice 
    
     The results of the test are shown in Table 7. As is seen in Table 7, all the active ingredients tested exhibited an antitumour activity. 
     
                       TABLE 7______________________________________Antitumour activity against Sarcoma - 180                   InhibitionCompound                ratio (I.R. %)______________________________________Sodium o-aminobenzoate-N--L-arabinoside                   44.4Sodium o-aminobenzoate-N--D-xyloside                   18.9Sodium o-aminobenzoate-N--D-glucoside                   22.4Sodium o-aminobenzoate-N--D-galactoside                   21.6Sodium o-aminobenzoate-N--L-rhamnoside                   57.4Sodium o-aminobenzoate-N--D-mannoside                   32.0Methyl o-aminobenzoate-N--D-mannoside                   59.0______________________________________Note:Amount of administration was 10 × 500 mg/kg p.o. 
    
     (4) ANALGETIC ACTIVITY 
     Determination by the mechanical stimulation method (by applying pressure) 
     Female ICR mice which showed a threshold value of pain of 50 to 80 mmHg when their tail base part was pressured by a pressure stimulation apparatus (made by Natsume Works, Japan) of Takagi and Kameyama were chosen as test animals, ten animals comprising a group. 
     After administering the active ingredient, the test was carried out as the time passes and the applied pressure and the time period until the animal showed a quasi-escaping reaction were determined to evaluate the analgeticactivity of the active ingredient. 
     The results are shown in Table 8. As is seen in Table 8, the pressure applied on animals when the animal showed the quasi-escaping reaction was higher in animals to which the active ingredient had been applied than in animals not administered, and the time period from the beginning to the time point when the animal showed the reaction was longer in animals administered with the active ingredient than in animals not administered. Thus, the analgetic activity of the active ingredient was confirmed. 
     Determination by the chemical stimulation method 
     The active ingredient was orally administered to a group (ten animals) of female ICR mice of age of 5 to 6 weeks, and after 30 min of the administration an aqueous 0.6% acetic acid solution was intraperitoneally injected into the mouse at a dose rate of 0.1 ml/10 g of body weight. The number of writhing motion which occurred in the mouse during 10 minutes after 10 minutes of intraperitoneal administration was recorded. The analgetic activity was evaluated from the writhing syndrome inhibiting ratio obtained by the following formula: 
     
         (1-T/C)×100=writhing syndrome-inhibiting ratio (%), 
    
     wherein 
     T: mean number of writhing syndrome in the group administered 
     C: mean number of writhing syndrome in the control group. 
     The results are shown in Table 9. As is seen in Table 9, every active ingredient of the pharmaceutical composition of the present invention showed analgetic activity. The above-mentioned process was carried out following the method of Kostet et al. (1959) 
     
                       TABLE 8______________________________________Analgetic activity by the mechanical stimulation method             Quasi-escape reaction             pressure at                     time until             (mm Hg) (sec)Compound            occurrence______________________________________Sodium o-aminobenzoate-N--L-               82        38arabinosideSodium o-aminobenzoate-N--D-               88        37xylosideSodium o-aminobenzoate-N--D-               88        36glucosideSodium o-aminobenzoate-N--D-               80        36galactosideSodium o-aminobenzoate-N--L-               79        40rhamnosideSodium o-aminobenzoate-N--D-               92        41mannosideMethyl o-aminobenzoate-N--D-               75        35mannosideControl             70        33______________________________________Note:Amount of administration, 1000 mg/kg p.o. 
    
     
                       TABLE 9______________________________________ Analgetic activity by the chemical stimulation methodCompound                 I.R. (%)______________________________________Sodium o-aminobenzoate-N--L-arabinoside                    43.1Sodium o-aminobenzoate-N--D-xyloside                    27.9Sodium o-aminobenzoate-N--D-glucoside                    24.5Sodium o-aminobenzoate-N--D-galactoside                    19.8Sodium o-aminobenzoate-N--L-rhamnoside                    52.1Sodium o-aminobenzoate-N--D-mannoside                    43.4Methyl o-aminobenzoate-N--D-mannoside                    40.5______________________________________Note:Amount of administration was 1000 mg/kg p.o. 
    
     (5) ANTIPYRETIC ACTIVITY 
     Following the method of Winter et al. (1961), a 20% suspension of beer yeast was subcutaneously injected to a group (consisting of 6 animals) of rats, and after 10 hours of fasting, the active ingredient was orally administered to the rats and their rectal temperature was determined. 
     The antipyretic activity is expressed by the ratio of inhibiting pyrexia due to beer yeast (I.R.%) at the time when the antipyretic activity of theactive ingredient is at its maximum according to the following formula: ##EQU1##wherein T: mean rectal temperature of rats to which the active ingredient was administered. 
     C 1  : mean rectal temperature of rats injected beer yeast, without the active ingredient. 
     C 2  : mean rectal temperature of untreated rats (control) 
     The results are shown in Table 10. As is seen in Table 10, all the active ingredients exhibited a considerable antipyretic activity. 
     
                       TABLE 10______________________________________Antipyretic activity                  Antipyretic activity                  (suppressing py-Compound               rexia) I.R. (%)______________________________________Sodium o-aminobenzoate-N--L-arabinoside                  35.7Sodium o-aminobenzoate-N--D-xyloside                  29.8Sodium o-aminobenzoate-N--D-glucoside                  15.6Sodium o-aminobenzoate-N--D-galactoside                  40.6Sodium o-aminobenzoate-N--L-rhamnoside                  66.6Sodium o-aminobenzoate-N--D-mannoside                  19.8Methyl o-aminobenzoate-N--D-mannoside                  25.0______________________________________ 
    
     (6) ANTIINFLAMMATORY ACTIVITY 
     (a) Carrageenin-edema inhibitory activity 
     Following the method of Van Arman et al. (1963), the active ingredient was forcibly and orally administered to each rat of a group consisting of 10 animals at the dose rate of 1000 mg/kg, and after one hour of the administration 0.1 ml of 1% suspension of carrageenin in physiological saline solution was injected to their right food pad. The volume of the foot pad was determined as time passes and the antiinflammatory activity was expressed by the ratio of inhibition of the swelling of the foot pad due to carrageenin by the active ingredient, using the determined value of1-4 hours from the injection and calculating by the following formula: 
     
         (1-T/C)×100=I.R.(%)=antiinflammatory activity 
    
     wherein 
     T: Mean value of volumes of planta in administered animals 
     C: Mean value of volume of the foot pad of control (not administered and then injected) 
     The results are shown in Table 11. As is seen in Table 11, all the ingredients tested showed the inhibitory activity against the edema causedby carrageenin. 
     (b) Antigranuloma activity 
     Following the method of Winter et al. (1963), two cotton wool pellets were implanted into the skin of back of each rat of a group consisting of 6 rats at the symmetrical positions having the median line as the axis of symmetry, the weight of one pellet being 30±1 mg. Oral administration of 1000 mg/kg/day of the active ingredient was carried out for consecutive7 days. On the 8th day, the granuloma formed in the rats was extirpated andweighed after drying. The antigranuloma activity expressed by the ratio of inhibition of the growth of the granuloma (I.R.,%) was calculated in a manner as shown in (6)(a), and the results are shown in Table 11. As is seen in Table 11, each active ingredient exhibited the inhibiting activityof growth of the granuloma. 
     (c) Antiexudation activity 
     Following the method of Baris et al. (1965), a volume of air was injected subcutaneously in the back of each rat of a group consisting of 6 rats to make a air pouch, and then 0.5 ml of 1% croton oil solution in sesame oil was injected into the pouch. The oral admnistration of 1000 mg/kg/day of the active ingredient was then began to continue for 5 days. On the 6th day, the amount of exudated liquid into the pouch was determined and the antiexudation activity expressed by the ratio of inhibitory activity to exudation was calculated in a manner as shown in (6) (a). The results are shown in Table 11. As is seen in Table 11, all the active ingredient tested exhibited the antiexudation activity. 
     (d) Antiadjuvant arthritis activity 
     Following the method of Fujiware et al. (1971), Mycobacterium tuberculosis suspended in liquid paraffin was injected subcutaneously into the right foot pad of each rat of group consisting of 6 rats. After 14 days of the injection, rats with similar volume of the foot pad were chosen to form groups (10 animals/group), each active ingredient was orally administered daily from the 15th day for consecutive 7 days. The volume of the foot padof rats was determined, and the antiadjuvant arthritis activity of each active ingredient was calculated as the ratio of inhibiting the swelling of the foot pad by using the formula shown in (6)(a). The results are shown in Table 11. As is seen in Table 11, all the tested active ingredient exhibited the antiadjuvant arthritis activity. 
     
                       TABLE 11______________________________________Antiinflammatory activity expressed                  *granu-  *exuda-                                  *arth-Compound       *edema  loma     tion   ritis______________________________________Sodium o-aminobenzoate-          26.0    6.3      10.5   22.7N--L-arabinosideSodium o-aminobenzoate-          4.6     10.1     18.0   19.0N--L-rhamnosideSodium o-aminobenzoate-          6.9     4.7      12.9   16.1N--D-galactosideSodium o-aminobenzoate-          20.1    12.6     17.8   28.6N--D-xylosideSodium o-aminobenzoate-          4.0     7.9      9.1    12.1N--D-glucosideSodium o-aminobenzoate-          4.8     11.8     7.1    17.8N--D-mannosideMethyl o-aminobenzoate-          21.5    10.7     15.4   11.5N--D-mannoside______________________________________Note:Amount of administration of the active ingredient = 1000 mg/kg 
    
     (7) BLOOD LIPID REDUCING ACTIVITY 
     Japanese male white rabbits were fed for about 3 months with solid feed (CR-1) containing 1% of cholesterol and those animals in which the increase of seral lipid component was confirmed were used as the model animals having experimental arteriosclerosis. 
     An aqueous solution of the active ingredient in distilled water was administered respectively at the dose rates of 30 and 300 mg/kg orally andafter the administration, blood specimen was collected as time passes from the auricular vein and the change of total cholesterol (determined by the enzyme method), phospholipid (determined by the enzyme method) and beta-lipoprotein (determined by turbidmetry) in the serum was observed. 
     The results are shown in Table 12. In Table 12, the values of serum cholesterol (mean value of 550 mg/dl), of phospholipid (mean value of 320 mg/dl) and of beta-lipoprotein (mean value of 2500 mg/kg) before administration were respectively subtracted from the respective values after 3 and 6 hours of the administration, and only the differences are shown, respectively. Therefore, the minus value shows the decrease and theplus value shows the increase of the respective values due to the administration. As is clearly seen in Table 12, generally, the active ingredients exhibited the activity of reducing the lipid components as compared to control. 
     
                                           TABLE 12__________________________________________________________________________Activity of reducing blood lipids                     Phospholipid                            beta-Lipoprotein                                     Cholesterol                Dose (mg/dl)                            (mg/dl)  (mg/dl)Compound             (mg/kg)                     3 hr                        6 hr                            3 hr                                6 hr 3 hr                                         6 hr__________________________________________________________________________Sodium o-aminobenzoate-N--D-mannoside                30   -40                        -40 -132                                -175 -98 -74                300  -43                        -59 -174                                -158 -120                                         -135Sodium o-aminobenzoate-N--D-glucoside                30   -27                        -40 -142                                -169 -96 -77                300  -33                        -43 -200                                -203 -72 -105Sodium o-aminobenzoate-N--D-galactoside                30   +11                        -27 -192                                -113 +3  -25                300  0  -30 -380                                -194 +10 -75Sodium o-aminobenzoate-N--L-arabinoside                30   -26                        -51 -213                                -179 +3  -100                300  -74                        -87 -300                                -202 -5  -210Sodium o-aminobenzoate-N--L-rhamnoside                30   -23                        -23 -148                                -197 -122                                         -115                300  -25                        -19 -220                                - 284                                     -180                                         -175Sodium o-aminobenzoate-N--D-xyloside                30   -19                        -47 -179                                -181 -50 -60                300  -23                        -64 -244                                -320 -100                                         -125Methyl o-aminobenzoate-N--D-mannoside                30   -24                        -32 -144                                -127 -181                                         -163                300  -37                        -39 -201                                -139 -230                                         -215Control                   0  -19 0   +3   +8  -4__________________________________________________________________________ 
    
     The formulation of the active ingredients to make the pharmaceutical composition of the present invention is described below. 
     In the case where the pharmaceutical composition is used as an antiinflammatory agent, it is possible to use the pharmaceutical composition in the form which is convenient to produce the effectiveness according to the kinds and the symptoms of the disease, and moreover, the active ingredient may be used by itself or may be used as mixtures combined with any diluent allowable in pharmaceutical process and with other medicines. 
     The pharmaceutical composition of the present invention is administered orally or parenterally and accordingly, the pharmaceutical composition of the present invention may take any form optionally for the oral or parenteral administration. 
     The pharmaceutical composition of the present invention may be offered as aform of unit administration. The form of the pharmaceutical composition of the present invention may be powder, granule, tablet, sugar-coated tablet,capsulated one, suppository, suspension, solution, emulsifiable concentrate, ampouled one, injection, etc. As a diluent, any one of solids, liquids and semisolids may be utilized, for instance, excipients, fillers, binders, wetting agents, disintegrating agents, surfactants, demulcents, dispersing agents, buffering agents, perfumes, preserratives, dissolution aids and solvents. Moreover, one or more than one of these adjuvants may be used in combination or in mixtures. 
     The pharmaceutical composition of the present invention may be formulated by any known method, and the amount of the active ingredient contained in the composition (preparation) is generally from 0.01% to 100% by weight. 
     The pharmaceutical composition of the present invention may be administeredorally or parenterally to human or animals, however, it is preferably administered orally. Sublingual administration is included in oral administration. Parenteral administration includes subcutaneous-, intramuscular- and intravenous injection and the injection by drop method. 
     The dose of the pharmaceutical composition of the present invention dependsupon the age, the personal difference and the state of disease, and whetherthe object is human or animal and accordingly, an extraordinary amount may be administered other the following dose: Generally, for human, the oral dose is 0.1-1000 mg/kg body weight/day, preferably 1-500 mg/kg/day and theparenteral dose is 0.01-200 mg/kg/day, preferably 0.1-100 mg/kg/day dividedinto 1-4 parts, one part being administered in one time. 
     The followings are the more detailed explanation of the formulation and theproduction of the pharmaceutical composition of the present invention in examples. 
     EXAMPLE 1 (Formulation) 
     10 parts by weight of one of the active ingredients of the present invention (Sodium o-aminobenzoate-N-L-arabinoside), 
     15 parts by weight of (heavy) magnesium oxide and 
     75 parts by weight of lactose were uniformly mixed and formulated into powder or granules. The powder is filled in capsules to be capsulated formulation. 
     EXAMPLE 2 (Formulation) 
     45 parts by weight of one of the active ingredients of the present invention (Sodium o-aminobenzoate-N-D-xyloside), 
     15 parts by weight of starch, 
     16 parts by weight of lacotse, 
     21 parts by weight of crystalline cellulose, 
     3 parts by weight of polyvinyl alcohol and 
     30 parts by weight of water were uniformely mixed, crushed and formulated, and then dried and shifted to be granules. 
     EXAMPLE 3 (Formulation) 
     Granules were prepared as in Example 2, except using sodium o-aminobenzoate-N-D-glucoside instead of sodium o-aminobenzoate-N-D-xyloside, and the mixture of 96 parts by weight of this granule and 4 parts by weight of calcium stearate was compression-formulated to be tablets 10 mm in diameter. 
     EXAMPLE 4 (Formulation) 
     94 parts by weight of one of the active ingredients of the present invention (Sodium o-aminobenzoate-N-L-rhamnoside), 
     6 parts by weight of polyvinyl alcohol and 
     30 parts by weight of water were mixed and the mixture was processed as in Example 2 to be granules. To 90 parts by weight of the thus processed granules 10 parts by weight of crystalline cellulose were mixed and the mixture was compression-formulated to be tablets 8 mm in diameter. The tablets were coated with syrup, gelatine and precipitated calcium carbonate to be coated tablets. 
     EXAMPLE 5 (Formulation) 
     0.6 part by weight of one of the active ingredients of the present invention (sodium o-aminobenzoate-N-D-galactoside), 
     2.4 parts by weight of a non-ionic surfactant and 
     97 parts by weight of physiological saline solution were mixed under heating and then the mixture was sterilized to be an injection. 
     EXAMPLE 6 (Production of o-aminobenzoic acid-N-L-arabinoside and its sodiumsalt) 
     A mixture of 2.3 g of anthranilic acid, 2.5 g of L-arabinose and 0.2 g of ammonium chloride was heated in 30 ml of methanol under a reflux condenser. After the reaction was over, crystals separated out when the reaction mixture was left at room temperature. The crystals thus obtained by filtering were washed with water, methanol and then with ether. The crystals were colorless needles or plates. Yield was 61.3%. 
     Thus obtained anthranilic acid-N-L-arabinoside was dissolved gradually intoan aqueous 1% sodium hydroxide solution containing in total the amount of calculated sodium hydroxide and after filtering, the solution was condensed under reduced pressure. The crystals which separated out by the addition of a large excess of acetone to the condensate was dehydrated anddried. Colorless crystals of sodium salt was obtained at the yield of 100%.The total yield from anthranilic acid was 61.3%. 
     EXAMPLE 7 (Production of o-aminobenzoic acid-N-D-xyloside and its sodium salt) 
     A mixture of 2.3 g of o-aminobenzoic acid, 2.5 g of D-xylose and 0.2 g of ammonium chloride was heated in 35 ml of ethanol under a reflux condenser.After the reaction was over, the reaction mixture was condensed under reduced pressure to half of the original volume, and after leaving at roomtemperature crystalline, needles separated out. After washing with water, methanol and then ether, the crystals were recrystallized from ethanol. Colorless needles were obtained at the yield of 74.6%. In the case where ammonium sulfate was used instead of ammonium chloride in the above-mentioned experiment, quite a similar result was obtained. 
     Thus obtained anthralinic acid-N-D-xyloside was gradually dissolved in an aqueous 1% sodium hydroxide solution containing the alkali in an calculated amount. After filtering and condensing the solution, a large excess of acetone was added to the condensate to obtain wet crystals. Colorless crystals were obtained after dehydration and drying at the yieldof 100% based on the anthranilic acid-N-D-xyloside. Total yield from anthranilic acid was 74.6% of theoretical. 
     EXAMPLE 8 (Production of o-aminobenzoic acid-N-D-glucose and its sodium salt) 
     A mixture of 4.6 g of anthranilic acid, 6.0 g of D-glucose and 0.5 g of ammonium chloride was heated in 40 ml of 95% ethanol under a refux condenser. After the reaction was over, the reaction mixture was condensedto about one third of the original volume and left in a refrigerator for a night to form of crystals. After collecting crystals by filtration of the reactant solution, and washing the filtered crystals with water, methanol and then ether and further recrystalizing the washed crystals two times from methanol, colorless crystals were obtained at the yield of 4.6%. 
     By dissolving thus obtained crystals into an aqueous 1% sodium hydroxide solution in stoichiometric proportions, and filtering the solution, then condensing the filtrate and at last adding a large excess of acetone into the condensate, crystals separated out from the acetonic solution. After dehydration and drying, colorless needle-like crystals were obtained at the yield of 100% of the anthranilic acid-N-D-glucoside. Total yield from anthranilic acid was 4.6%. 
     EXAMPLE 9 (Production of o-aminobenzoic acid-N-D-galactoside and its sodiumsalt) 
     A mixture of 2.4 g of anthranilic acid, 3.0 g of D-galactose and 0.2 g of ammonium chloride was heated in 30 ml of 95% ethanol under a reflux condenser. After the reaction was over, the reactant was condensed to one half of the original volume under reduced pressure and left at room temperature to separate crystals. After filtration of the reactant and washing the collected crystals with water, methanol and then ether and further recrystallizing from 95% ethanol, colorless needle-like crystals were obtained at the yield of 16.4%. 
     Thus obtained crystals of anthranilic acid-N-D-galactoside were dissolved in 1% sodium hydroxide solution in stoichiometric proportions, and after filtration of the solution, and condensation of the filtrate to one half in volume, a large excess of acetone was added to the condensate. Thus separated crystals were dehydrated and dried. Colorless crystals were obtained at the yield of 100% based on the anthranilic acid-N-D-galactoside and at the total yield of 16.4% based on anthranilic acid. 
     EXAMPLE 10 (Production of o-aminobenzoic acid-N-L-rhamnoside and its sodiumsalt) 
     A mixture of 2.3 g of anthranilic acid, 2.8 g of L-rhamnose and 0.2 g of ammonium chloride was heated in 25 ml of methanol under a reflux condenser. After the reaction was over, the reactant was left at room temperature to separate crystals. After filtering the reactant and washingthe collected crystals with water, methanol and then recrystallizing the washed crystals from 50% methanol, colorless needle-like crystals were obtained at the yield of 9.8%. 
     Thus obtained anthranilic acid-N-L-rhamnoside was slowly dissolved in 1% sodium hydroxide solution in stoichiometric proportions. After filtering the solution and condensing the filtrate to one half of the original volume, a large excess of acetone was added to the condensate to obtain crystals. By dehydrating and drying the wet crystals, colorless crystals were obtained at the yield of 100% based on the anthranilic acid-N-L-rhamnoside, the total yield based on anthranilic acid being 9.8%. 
     EXAMPLE 11 (Production of methyl o-aminobenzoate-N-D-mannoside) 
     A mixture of one gram of methyl anthranilate and one gram of D-mannose was heated in 10 ml of ethanol in the presence of 0.1 g of ammonium chloride for about one hour to effect condensation. After the reaction was over, the reactant was left at room temperature to separate crystals. The crystals were recrystallized from 95% ethanol to be colorless crystals with a yield of 60%. 
     EXAMPLE 12 (Production of o-aminobenzoic acid-N-D-mannoside and its sodium salt) 
     A mixture of 2 g of anthranilic acid, 3 g of D-mannose was heated in 10 ml of ethanol in the presence of 0.2 g of ammonium chloride under a reflux condenser, in a waterbath at 95° to 96° C. After a while, thick crystals separated out. After collecting the crystals by filtration and washing the crystals with water and methanol thoroughly, the crystals were recrystallized from methanol to be colorless needles. The yield was 53.0% based on anthranilic acid. 
     Thus obtained anthranilic acid-N-D-mannoside was slowly dissolved in 1% sodium hydroxide solution in stoichiometrical proportions. Undissolved matter, if any, was removed by filtration, and the solution (or the filtrate) was condensed under reduced pressure and then a large excess of ethanol was added to the condensate. The crystals separated were collectedby filtration and they were dehydrated and dried to give colorless crystalsat a yield of 100% based on the anthranilic acid-N-D-mannoside and at the total yield of 53.0% based on anthranilic acid.