FR2485926A1 - PHARMACEUTICAL COMPOSITION COMPRISING AN AMINOBENZOIC ACID DERIVATIVE TO REGULATE PROSTAGLANDINS - Google Patents

Abstract

PHARMACEUTICAL COMPOSITION INCLUDING A DERIVATIVE OF AMINOBENZOIC ACID TO REGULATE PROSTAGLANDINS. THIS COMPOSITION IS CHARACTERIZED IN THAT IT INCLUDES: A. A COMPOUND RESPONDING TO THE GENERAL FORMULA: (CF DRAWING IN BOPI) IN WHICH R DESIGNATES A MEMBER OF THE GROUP CONSTITUTED BY THE RADICALS FORMED BY REMOVING OH IN POSITION 1 (ALPHA) OR 1 (BETA) OF ARABINOSE, XYLOSE, RHAMNOSE, GLUCOSE, GALACTOSE, MANNOSE AND FRUCTOSE, AND R IS A HYDROGEN ATOM, A CAC ALKYL GROUP OR A PHARMACEUTICALLY ACCEPTABLE METAL, AND B. A PHARMACEUTICALLY ACCEPTABLE CARRIER OR DILUENT FOR THIS.

Description

The present invention relates to a composition

  pharmaceutical regulation for the regulation of

  glandins in mammalian organisms, which comprises (a) as an active ingredient, an aminobenzoic acid derivative having the general formula below COOR-coolrs- ',

I NH-R

  wherein R denotes a member selected from the group consisting of radicals formed by removing OH at position 1 (alpha) or 1 (beta) from arabinose, xylose, rhamnose, glucose, galactose, mannose and fructose, and R2 is a hydrogen atom,

  a C1-C4 alkyl group or a pharmaceutical metal

  acceptable and (b) a pharmaceutical carrier or diluent

  maceutically acceptable for it.

  A group of compounds known generically as prostaglandins (hereinafter referred to as

  PGs or PGs (plural) have recently taken

  because of their varied physiological functions in mammalian organisms. However, as some of the PGs have short half-lives or are

  unstable, their use as pharmaceuticals

  ticks posed problems. There is a process for

  regulate PGs formed in mammalian organisms.

  fires, for example the administration of aspirin (see

Nature, 239: 33-34, 1972).

  However, aspirin is a cyclooxygenase inhibitor that participates in the early stage of PG metabolism, and as a result, aspirin

  should be considered the blocker of all PGs.

  In other words, aspirin should be considered as a regulator of the production of all PGs, and it has no activity of increasing the production of a certain PG or PGs, and Consequently, there is of course a limit to

  the activity of aspirin as a regulator of PGs.

  In addition, aspirin has side effects, it

  causes gastroenteric disturbances in mammals

  fers, its long-term administration therefore raises

problems.

  Having searched for a compound capable of regulating PGs without any side effects as far as possible, the Applicant has found that a series of amino-benzoic acid derivatives represented by general formula (1) above , have effective regulatory activity of PGs and it has

resulted in the present invention.

  The above derivative of amino-benzoic acid

  intended for use as an active ingredient in accordance

  according to the present invention is a chemical compound

  and physically known. Inoue et al have described the chemical synthesis of this compound (see J. Agr.Chem.Soc.Sk., Vol 25, pp. 59-63 and 291-293 (1951), and Chemical Abstracts Vol 48, Columns 2001d and 2001i (1954)) and certain physical properties of the compound (see J. Agr Chem Chem Soc Japan, Vol 26, p.329 - 331 (1952)), and Chemical Abstracts Vol. 48, Column 2003a

(1954)).

  However, in the references above, nothing is indicated regarding the properties

  physiological or pharmaceutical of the above compound.

  In addition, no indication has been found so far regarding the physiological and / or pharmacological properties of the compound of the formula

general <1).

  In one aspect, the invention relates to a pharmaceutical composition capable of regulating prostaglandins PGs in mammalian organisms, based on the novel medical use of the chemical compounds represented by the general formula (1) above: In the drawings Fig. 1 shows the effect of the substance of the invention on platelet aggregation by arachidonic acid; FIG. 2 shows the effect of the substance of the invention on platelet aggregation by adenosine diphosphate (ADP); FIG. 3 shows the effect of the substance of the invention on collagen aggregation by collagen; FIG. 4 shows the effect of the substance of the invention on platelet aggregation by epinephrine; FIG. Fig. 5 shows the effect of the substance of the invention on platelet aggregation by ristocetin; Fig. 6 shows the effect of the substance of the invention on platelet aggregation by thrombin; 7 shows the effect of the substance

  of the invention on the rate of monoDhosDhate of

  nosine cyclic (cyclic AMP) in the wafer, FIG. 8 shows the effect of the substance of the invention on the production of malonedialdehyde (MDA) in the wafer,

  FIG. 9 shows the effect of indom-

  thai on the hypotensive action of the substance of the invention; 10 shows the effect of the substance of the invention on the cyclic AMP level in cancer cells; FIG. 11 shows the effect of the substance of the invention on the production of prostaglandins 12 (PGI2) in 3T3 fibroblasts; FIG. Fig. 12 shows the effect of the substance of the invention on the prostaglandin F 2 (PGF 2) level. Fig. 13 shows the effect of the substance of the invention on the prostaglandin E (PGE) level. 14 shows the effect of the substance of the invention on the level of PGs in the plasma of

  rats administered the substance of the invention.

  tion, FIG. Shows the effect of the substance of the invention on the prevention of platelet aggregation induced by ADP, and FIG. 16 shows the effect of the substance of the invention on reducing the level of PGE in the cerebrospinal fluid after it has been elevated by

pyrogenic microbes.

  The active ingredient of the pharmaceutical composition

  According to the invention, hereinafter the substance of the invention is a compound corresponding to formula (1) below:

COOR

I

> NH'àNH1

  wherein R 1 - denotes a member selected from the group consisting of radicals formed by removing OH at position 1 (alpha) or 1 (beta) from arabinose, xylose, rhamnose, glucose, galactose, mannose and fructose, and R2 is a hydrogen atom,

  a C1-C4 alkyl group or a pharmaceutical metal-

acceptable.

  SR 1367 JA MdT Since the substance of the invention, as will be explained hereinafter, has a toxicity

  extremely low acute oral level, has no muta-

  genicity, does not affect the cellular and humoral immunity of the host and does not disturb the bacterial flora.

  gastrointestinal tract because of the lack of activity

  anti-bacterial, it can be given orally in the long term without risk of mutagenesis

or allergy.

  Accordingly, the substance of the present invention is a pharmaceutical product of great

work security.

  In the general formula (1) above of the substance of the invention, R1 is abbreviated to a radical of a monosaccharide, and the monosaccharide may be either in the D form, or in the L form, or an alpha anomer, either a beta anomer or a mixture

  anomers. Consequently, the substance of the in-

  vention may itself be either an alpha-anomer or

  a beta-anomer, a mixture of the anomers.

  In the general formula (1) above, R2 is a hydrogen atom, a pharmaceutically acceptable metal such as Na, K, 1/2 Ca, 1/2 Mg, 1/3 Al or an alkyl group such as a methyl, ethyl, propyl group

and butyl.

  The process for preparing the substance of the invention may be described by way of example as follows:

  A mixture of 5 g of p-amino acid is heated

  benzoic acid, 5 to 6 g of a monosaccharide such as L-arabinose, lxylose, L-rhamnose, D-glucose, D-galactose,

  D-mannose and D-fructose, and 0.1 to 0.5 g of chlorine

  ammonium chloride, magnesium chloride, formic acid, acetic acid or hydrochloric acid in 90 ml of 95-100% ethanol or pure methanol,

  under a reflux condenser, to initiate condensation.

  When the reaction is complete, the reagent is left at room temperature or in a cold place and the crystals which separate are collected by filtration. They are washed with water, with ethanol or with ethyl ether and then recrystallized from methanol, ethanol or an aqueous solution of

methanol or ethanol.

  To replace the hydrogen atom of the carboxyl group of the compound thus prepared with a base, it is preferable to follow the known method. The compound, N-pyranoside of p-aminobenzoic acid is dissolved in an aqueous ethanolic solution and added to

  the solution a mineral salt to perform the substitution.

  The physical properties of certain substances of

  the invention (the active ingredient of the pharmaceutical composition

  The substances of the invention shown in Table 1 are hereinafter referred to as N samples if at 11.

TABLE 1

  Physical properties of active ingredients

  Sample - Power Point Max.

  Name of the substance of the invention Elemental rotary fusion (%) UV absorption N (C) Ia D20 C: H: N (millimicron) 1 ND-galactoside of sodiu o-aminobenzoate 157 - 163 9 48,5 : 5.024-4 317, 248, 215 (decap) in water

(48, 6: 5.0: -4.4)

152 - 162 + 54 51.0: 5.4: 4.9

  2 N-L-rhamnoside of sodium o-aminobenzoate (decomp.) In water (51, 1: 5.2: 46) 320, 249, 215

(51, 1: 5.2: 416)

-. -2 48.5: 5.0: 4.2

  3 N-D-mannoside of sodium m-aminobenzoate 130 - 14 +5 (48.6: 5.0: 4.4) 274

- 196 - 2 46.1: 5.5: 4.0

  4 ND-mannoside of sodium p-aminobenzoate 185 - 196 - 2 46,15,54,0 274 (decxxp) in water (46.0: 5.3: 4.1) ND-glucoside of p-aminobenzoate Sodium aminobenzoate 145 - 160, 45.8: 5.2: 4.3 274 in water (46.0: 5.3: 4.1) -M r Co Ln% O ru O TABLMU 1 (Continued) 6 n-galactoside of sodium p-aminobenzoate 150 - 162 + 46.0: 5.6: 4.1 275 in water (46.0: 5i3: 4.1) 7 N-p-aminobenzoate d-xyloside Sodium 149 - 158 O 49't3: 4; 9: 48 274 in water (49, 5: 4,8: 4,8) 8 NL-rhamnoside of sodium p-aminobenzoate 173 - 178 + 80: 5f14 , 8273 (deoorp) in water (51.1: 5.2: 4.6) 9 N-fructoside of sodium paminobenzoate 180 - 205 - 4 48.8: 4.9: 4.3 290 in water (48.6: 5, 0: 4; 4) NL-arabinoside of sodium p-aminobenzoate 163 - 173 - 41 50.0: 4.2: 4.7 274 (decapped) in water (49.degree. 8: 4.2: 4., 8) Amino-benzoate amino-benzoate of 177 - 178,54 in 49,1: 6,1: 4,3-n-3-mannoside of o-aminobenzoate methyl 177 - 178 - 54 in 330, 251 1 ethanol (48., 1: 6., 6: 4, O) Note: * Figures in parentheses are the theoretical values of C, H and N (%) rJ Co Ln m0 * 1 1) Melting point: It is determined using a micro-apparatus for determining the melting points produced by the

Yanagimoto Works, Japan.

  2) Rotatory power: It is determined using a Model OR-50 direct reading polarimeter manufactured by the Yanagimoto Works, Japan, with a thickness of 50 mm of an ethanolic aqueous solution of the acidic active ingredient and an aqueous solution of sodium salt of a

acidic active ingredient.

  3) Molecular Composition: The elemental analysis was carried out using a CHN-Coder Model MT-2 manufactured by the

Yanagimoto Works, Japan.

  4) Ultraviolet Absorption Spectrum:

  It is determined using a spectrophotometer

  Recorder Photometer Model PS-3T made by

  Hitachi Works, Japan, on an aqueous solution

  nolic of the active ingredient acid and on an aqueous solution of the sodium salt of the active ingredient acid

of the drug.

  The following will be given-the physio-

  of the substances of the invention described in the following order: (1) acute toxicity, (2) activity

  antimicrobial, (3) mutagenicity, (4) intra-

  dermal type, (5) anti-

body and (6) effects on the stomach.

  (1) Acute Oral Toxicity: Acute oral toxicity of the substances of the invention was examined using ICR-JCL mouse groups and forcibly administering each of the representatives of the substances of the invention as an oral aqueous solution in distilled water in the form of an aqueous suspension also in distilled water at predetermined doses, respectively. After observing the potential toxicological symptoms for 6 days, and recording mortality to the seventh day after administration, the LD50 (acute oral) was calculated by the Litchfield-Wilcoxon method, and autopsy on dead mice and on

  live mice to get information.

  Table 2 gives the results obtained.

  It can be seen that the LD50 (acute oral) of the representatives of the substances of the invention is very high, which shows that it has an extremely low toxicity.

TABLE 2

  Acute oral toxicity of representative substances Echan-. Ncam of the substance of the invention LD50 (g / kg)

p.o.

  1 N-D-galactoside of sodium o-aminobenzoate 6.10 2 N-L-rhamnoside of sodium o-aminobenzoate 12.50 3 N-D-mannoside of. sodium m-aminobenzoate> 10 4 Na-mannoside of sodium p-aminobenzoate> 10 N-D-glucoside of sodium p-aminobenzoate> 15 6 ND-Galactoside of sodium paminobenzoate 14.55 5-N-amylbenzoate p-aminobenzoate Sodium 11,75 8 NL-rhamnoside sodium p-aminobenzoate 12.80 9 N-D-fructoside sodium p-aminobenzoate> 10 NL-arabinoside sodium paminobenzoate 10.80 11 o-Aminobenzoate ND-mannoside > 7.5 In addition, no abnormal observations were made at autopsy on dead animals and

living animals.

  (2) Anti-microbial activity: The substance of the invention was dissolved in distilled water in a series of double dilution systems. These diluted solutions were mixed with agar-agar medium in nine times their volume and the mixture was poured into a petri dish. A heart-agar infusion medium and a medium were used for the bacteria. Sabouraud agar-agar for mushrooms. After being striped with preculture, the innoculated plates were incubated at 37 ° C. for 20-24 hours for the bacteria and at 25 ° C. for 3-7 days for the mushrooms, and then the growth was examined. The following microorganisms were used to determine

  the antimicrobial activity of the substance of the in-

  Pseudomonas aeruginosa IAM 1514 Escherichia 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 The above tests showed that none of the active ingredients tested had

  growth inhibition of all micro-organisms

at the concentration of 1 mg / ml.

  (3) Mutagenicity: In a first stage, the substances of the invention were subjected to the "rec" test (i), and in a second stage they were subjected to a reversion test (ii). (i) A strain of Bacillus subtilis M 45, a recombinant repair defect, and a wild strain of Bacillus subtilis H 17 retaining recombinant repair activity were innoculated to make their own trait

  crossed on a plate of agar culture

  B-2 agar (prepared 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 pH 7.0). Then placed on the surface of the agar plate was a paper disc 8 mm in diameter, having absorbed 0.04 ml of an aqueous solution of the substance of the invention in distilled water, way to cover the starting point of the aforementioned traits of bacterial culture. The innoculated B-2 agar plate is stored at 37 ° C. overnight and the length of the region whose growth has been inhibited is measured. As a negative control, Kanamycin is used and Mytomycin C as a positive control. The results of the "rec" test are given in FIG.

Table III.

  (ii) TA 98 and TA 100 strains (both require histidine) of Salmonella typhimurium

  were used for the reversion test.

  In 2 ml of a soft agar culture medium (the medium itself contains 6 g of sodium chrolide and 6 g of agar-agar in 1000 ml of distilled water)

  to which a tenth of its volume of a solution has been added.

  0.05 mM biotin and 0.5 mM histidine, 0.1 ml of the bacterial suspension and 0.1 ml of an aqueous solution of the substance of the invention are mixed and the mixture is stirred. the agar-agar culture medium minimum. After two days of incubation at 37 ° C., the number of revertant colonies is counted. As a positive control, furylfuramide (AF-2) is used. The results of the test of

  reversion are given in Table 4.

  As shown in Table 3, the substances of the invention exhibit low mutagenicity only at high concentration (5000 micrograms / disc). Table 4 shows that the rate of occurrence of the mutation by the substance of the invention shows no difference from the tem-

  no substance has been added, even to a concentration

  high level (5000 micrograms / plate). These observations show that the substance of the invention is of a

  safe use with regard to mutagenicity.

TABLE 3

  Result of the test "rec" Sample length, of inhibition - Difference of

  Name of the substance of the invention Concentration of growth zones

ion N sance (mu)

M45 H 17

  (μg / disc) (mm) (m) 1 O-O-O-o-aminobenzoate n-galactoside sodim% SO5dium 000 6 1 5 NL-o-aminobenzoate NL-rhamntoside sodium 5 000 6 2 4 ND-mannoside of 500,000 3 sodium m-aminobenzoate 5 000 7 1 6

N 5000 0 0

  4 N-D-nannoside of p-aminobenzoate 500 O sodium 5 000 O 4 -4. cs N-Dglucoside sodium p-aminobenzoate 000 o O o S, - ,, -, -, i ,,,,,., ..,

500 0 0 0

  6 N-D-galactoside p-aminobenzoate 500 O sodium000 O O O

000 0 0 0

  7 N-D-xyloside of 50 O 7 sodium p-wminobenzoate 5 000 O O O 8 N-Lrhamnoside of 500 o O 8 sodium p-aminobenzoate

_ _ _ _ 5 OQQ 0 0 O

  9 N-D-fructoside p-aminobenzoate 500 O O. sodium

000 0 0 0

  500-O-sodium p-aminobenzoate N-L-arabinoside

000 0 0 0

5000 O O O

  ND-mannoside of o-aminobenzoate 500,000 mthy1e 5000 7 3 4 Kanamy5ine 10 5 4 0 Mit-amyine C 0.05 12 2 10 e 1 Cu x% O (% 4 uLn P-4 _ _ Note: m Difference = length of the inhibition zone of M 45 minus length of the

H 17 inhibition zone.

TABLE 4

  Results of the reversion test 0% t '> 4% O Echan- Number of colonies of

ticlon-

  Name of the substance of the invention Referring concentration <number (g / plate) per plate) Na

TA 100 UA 98

  1 Na-galactoside of sodium o-aminobenzoate 5 000 151 6 2 N-Lrhamnoside of sodium o-aminobenzoate 5 000 61 9 3 ND-mnannoside of sodium half-aminobenzoate 5 000 90 6 4 ND-mannoside of sodium p-aminobenzoate 5 000 138 5 Sodium aminobenzoate ND-glucoside 5 000 134 Table 4 (cont.) sodium p-aminobenzoate ND-galactoside 000 7 sodium p-aminobenzoate ND-xyloside 5 000 58 4

j,, H. ,, ,,,,.

  Sodium p-aminobenzoate NL-rhamnoside 5 000 73 4 9 sodium p-aminobenzoate ND-fructoside 5 000 95 5 sodium paminobenzoate NL-arabinoside 5 000 95 8 11 o-aminobenzoate ND-mannoside methyl 5 000 '51 3 Furylfuramide 0.1 911 167 Control (no addition) - 149 13-m. (4) Intracutaneous Delay-type Reaction For the effects of the substances of the invention on cellular immunity, the footpad reaction test was performed using as animal smarter ICR-JCL and

  as antigens of sheep erythrocytes.

  A mouse was first sensitized by injecting 0.2 ml of an aqueous suspension

  10% sheep erythrocytes in physiological serum

  the caudal vein and, seven days after the first sensitization, 0.05 ml of a 40% aqueous suspension of sheep erythrocytes in physiological saline was injected into the plantar pad.

  for a second sensitization. The thickness of the neck

  Plantar sinet was determined the next day. The D-

  The substance of the invention was administered at a dose of 250 mg / kg / day once daily for five consecutive days focusing on the day

  o the first sensitization was done.

  We found that the increase in thickness

  of the plantar pad of the mice that received

  the administration of the substances of the invention did not

  no significant difference from the increase

  mentation in the group of mice that received no

  administration of active ingredients.

  (5) Antibody production activity To determine the effects of the substances of the invention on humoral immunity, the haemagglutination test was carried out using mice

  ICR-JCL sensitized with sheep erythrocytes.

  A mouse was sensiiblized by injecting 0.2 ml of a 10% aqueous suspension of sheep erythrocytes into physiological saline by the tail vein and, seven days after sensililization, the blood of the mouse was sampled for the hemagglutination test for the determination of anticopre production activity. The substance of the invention was administered for five consecutive days focusing on the day of sensitization,

  intraperitoneally at a dose of 250 mg / kg / day.

  No significant difference was observed in the agglutination titre between the group that received the substance of the invention and the control group. (6) Effect of the substance of the invention on the stomach: 5 g of the substance of the invention (each of NOS1, 2, 3, 4, 5, 6, 7, 8, 9, 10 and 11) and aspirin to each Beagle dog with a body weight of 9 to 12 kg in the form of an aqueous suspension in distilled water adjusted to pH 3, and 90 minutes after administration, examined the hemorrhagic state

  of the dog's stomach. Sto hemorrhage was observed.

  only in dogs given aspirin, however, no stomachable haemorrhage was found in dogs

samples ncs 1 to 11.

  The description below is the summary of

  peculiarities of the substances of the invention

  tion, a more detailed description being given in

the examples:

  (1) Prostaglandin regulation The substance of the invention relates to the regulation of PGs such as PGA, PGB, PGC, PGD, PGE, PGF, PGG, PGH, PGI, TXA and TXB and their metabolic products. In addition, the substance of the invention

  is not limited to regulating any of the PGs

  above, but it also regulates several species

of PGs.

  The regulatory function of the substance of the invention on the production of PGs and their metabolites is indicated by the following facts; (a) suppression of platelet aggregation: It is known that platelet aggregation is predominantly induced by PGG2, PGH2 and TXA2 and it has been confirmed that the substance of the invention has

  a function of suppressing the aggregation of plaquet-

tes (see examples 2 and 5).

  (b) Increasing cyclic adenosine monophosphate: It is known that cyclic adenosine monophosphate (cyclic AMP) is closely related to PGs, in particular PGD, PGE and PGI, and the substance of the invention has a function of increasing the level of cyclic AMP in platelets and

culture (see examples 3, 6 and 8).

  (c) suppression of malondialdehyde increase:

  It has been confirmed that the substance of

  has an inhibitory function on the production of malondialdehyde (MDA) in platelets, MDA being known as a metabolic product of PGs (see

example 4).

  (d) acceleration of PGI production: It has been confirmed that the substance of the invention has an accelerating effect of PGI2 production in the cells used in vitro (see

Example 7).

  (e) effect in relation to the production of certain PGs: In accordance with the experimental results on PG metabolism in vitro using arachidonic acid as starting material, it has been shown that the substance of the invention is related to the production of PGD2, PGE2, 6-keto-PGFla and

PGF2_a (see example 1).

  (f) effect on the biosynthesis of PGE and

jPGF 2-a: -

  It has been shown by an in vitro culture experiment that the substance of the invention has an effect on the biosynthesis of PGE and PGF2a (see

Examples 9 and 10).

  (g) Inhibition of tumor cell proliferation: Examination of cell proliferation

  tumors and the inttatumoral cellular PGE level.

  on experimental animals carrying tumors having received the substance of the present invention confirmed the inhibition of proliferation of tumor cells

  and also the increase in the level of PGE in

  Tumors of the animals that received the substance

  of the invention (see Examples 1, 12, 13 and 14).

  (h) enhancement effect of pigment transfer In an experiment in which the substance

  of the invention has been administered to experimental animals.

  Tumor-bearing disease and in which the degree of transfer of a dye into the tumor tissue of the animal was examined, an enhancer effect on the transfer of the exogenous dye was confirmed, suggesting the vasodilator effect of substance of

the invention (see Example 15).

  (i) inhibition of tumor metastases In an experiment in which the substance

  of the invention has been administered to experimental animals.

  before and after cell transplantation:

  to animals, an inhibitory effect has been observed.

  metastasis of transplanted tumor cells

(see example 16).

  (j) reduction of 6-keto-PGF: In tumor bearing animals, it has been observed that the level of 6-keto-PGFl-x in their plasma is remarkably increased compared to that of animals. However, it has been confirmed that the increased level is brought back to normal by administration of the substance of the invention (see Example 17) In general, PGs have been found in several organs of the whole body. , and they have

  close relations with the functions of the organs.

  In addition, it is known that the physiological functions

  and pharmacological aspects of PGs cover an extremely

  vastly That is, the PGs act primarily

  on the expansion and contraction of

  blood vessels with regard to their pharmacological action on the circulatory system, PGE and PGI being stronger

  for dilatation, and the IXA contracting on the contrary the artery. These functions lead to an increase or reduction of blood pressure and are a remission to stenocardia and

arrhythmia.

  Although prolonged platelet aggregation causes arteriosclerosis, cerebral infarction, myocardial infarction and cerebral apoplexy, PGs have pronounced effects on platelets. That is, DMP, EMP and PGI

  have an antagonistic function against the aggregating action

  platelet, and TXA2, PGG2, PGH2 and PGE2 have a priming or acceleration action on aggregation. As a result, it is clear that the treatments and prevention of the above diseases would be made possible by an appropriate adjustment

PGs.

  In addition, in diabetes it has been reported that the production of PGI2 in the diabetic patient's vascular system is limited, or that the PGE and PGF2 content is high, and the relationships between this

  disease and PGs have been progressively clarified.

  The functions of PGs in the respiratory system

  have also been studied and it is well known

  that PGE reduces the resistance of air ducts.

  This suggests an anti-asthma, acceleration of breathing, antitoxic and anti-sputum effect. On the other hand, as a function of immunity, the suppressive action on the release of the slow-reacting substance of anaphylaxis has been found in PGI2 and its metabolites. PGs

* also have an anti-allergic action, an anti-allergic action

  anaphilactic and it is to be presumed that they will be effective in the treatment of various autoimmune diseases, especially incurable diseases

what constitutes rheumatism.

  It is well known that indomethacin, a medi-

  anti-inflammatory drug currently in commerce,

  is related to cyclooxygenase in the metabolic pathway

  that PGs and that several species of PGs concern the inflammatory action. In summary, an anti-inflammatory effect can be expected by the regulation of

metabolism of PGs.

  On the other hand, PGs exert a marked effect on the digestive system: PGE and PGI

  suppress the secretion of gastric juice, and they do not

  therefore, to present an anti-

  ulcer. This action would be particularly strong

for the PGI.

  With regard to the kidney, the PGI would present

  mainly a diuretic action. Among the anti-agents

  commercial hypertensives, the place occupied by diuretics is still important, so that

  the increase in PGI2 production makes it interesting

  as a diuretic and as an antihypertensive agent.

  The pharmacological action of PGs on the system

  genital is also well known, and the PGs intervene

  to facilitate the action of uterine movement and uterine tension or vice versa the suppresive action of uterine tension. These functions are the basis of the evaluation of the PGs as an acceleration agent for delivery, as a contraceptive and as a termination agent for pregnancy. In addition, the action of PGs as a medicine

  psychoneural is under study. In the brain cells

  a large amount of PGD is distributed, and we are now looking with a keen interest

  possible between epilepsy and PGD.

  It is known that the peroxidation of lipids is accelerated by aging and arteriosclerosis, and that consequently the activity of the enzyme synthesizing PGI is suppressed. In addition, it has also been reported that this PGI has a stabilizing action on the lysosome membrane. These facts show the value of PGI as a peroxidized anti-lipid agent, and lead to the possibility of preventing radiation disorders and aging - Recently, PGs have - received a

  special attention about cancers. In part-

  In particular, several studies have been carried out on PGD, PGE and PGI in the above field and it has been discovered that these PGs relate to the suppressor effect on the proliferation of cancer cells, the normalization of cancer cells, the metastases of cancer, etc. These facts go on to suggest the possibility of suppressing cancerous proliferation

  and to avoid the metastasis of cancer.

  As has been indicated, the pharmaceutical functions

  Many PGs cover different areas in different ways, and it is not an exaggeration to say that PGs are involved in some way in all diseases. Therefore, since the substance of the invention regulates PGs, PGs can be expected to be useful in preventing and treating

diseases above.

  For example, with respect to its antitumor function, as described above, anti-tumor activity against L cells has been found for PGE (DR Thomas et al., Experimental Cell Research 84, 40-46). (1974y), and it was found that the increase and decrease of DMP were

  related to the formation of metastases and the proliferation

  tumors (F.A. Fitzpatrik et al., Proc Natl Acad Sci, USA, 76 (4) 1765-1769 (1979)) and that

  PGI was suppressive to proliferation

  tumors. However, this information only shows that each prostaglandin has activity

  anti-tumor is one of these physio-

  logical. On the other hand, as will be seen in Example 1, the Applicant has found that the substance of the invention has the effect of modifying the amount of PGD and PGE in the cells, i.e. it has the effect of regulating the PGs which differ from each other in the same cell. In addition, as shown in another example, it is clear that the substance of the invention has a suppressive or accelerating action on TXA and PGI, and this clearly shows that the substance

  invention is not limited to modifying a prostitu-

  single glandin, but also modifies simultaneously

various PGs.

  Such a substance modifying several species of prostaglandinessimultaneously was not known until now, and this property can be

  considered to be peculiar to the substance of the in-

vention.

  The formulation of the active ingredient to prepare the composition will now be described below.

pharmaceutical composition of the invention.

  When the pharmaceutical composition is used as a regulating agent for PGs, it is possible to use the pharmaceutical composition in the form to obtain efficacy depending on the nature and symptoms of the disease, and furthermore it is possible to use the active ingredient as or in admixture with any acceptable diluent in the pharmaceutical processes and with

other drugs.

  The pharmaceutical composition of the invention

  administered orally, or parenterally, and therefore

  it can take any desired form

  suitable for oral or parenteral administration.

  The pharmaceutical composition of the invention may be presented in unit dose form. It may be in the form of a powder, granules, tablet, sugar-coated tablet, capsule, suppository, suspension, solution, emulsifiable concentrate, ampoule, injectable ampule, etc. As the diluent, any solid, liquid or semi-solid diluent, for example excipients, binders, wetting agents, disintegrating agents, surfactants, demulsifiers, dispersants, buffering agents, perfumes may be used. , preservatives, dilution adjuvants and solvents. In addition, one or more of these adjuvants can be used in combination

or in mixture.

  The pharmaceutical composition of the invention can be formulated by any known method, and the proportion of the active ingredient (substance of the invention) contained in the composition (preparation)

  is in general from 0.01% to 100% by weight.

  The pharmaceutical composition of the invention may be administered orally or parenterally to humans or animals, but it is preferable to

  administer it orally. Sublime administration

  guale is included in the oral administration. The D-

  parenteral administration includes subcutaneous injection

  cutaneous, intramuscular and intravenous, and injection

by the drop method.

  The dose of the pharmaceutical composition of the invention depends on the age, the personal differences, and the state of the patient, it also varies according to whether it is a human being or an animal, but it is generally in the following areas: for men, the oral dose is 0 to 1. 1000 mg / kg body weight / day, preferably 1 to 500 mg / kg / day and the parenteral dose is 0.01-200 mg / kg / day, preferably 0.1-100 mg / kg / day, divided into one to four parts, only one part being administered at a time. The following is a more detailed explanation of the formulation, production and

  physiological activities of the pharmaceutical composition

  of the invention, in the form of examples.

  FORMULATION EXAMPLE 1 (in the following, the parts are by weight) The following constituents are uniformly mixed, the mixture is pulverized or granulated

  to transform it into a pulverulent formulation

  rulente of average size less than 350:

(1) N-D-galactoside of p-amino

  sodium benzoate 10 parts (2) heavy magnesium oxide 15 parts (3) galactose 75 parts The formula thus prepared is used as

what or after encapsulation.

EXAMPLE OF FORMULATION 2

  The components below are uniformly mixed, sputtered and converted into wet granules. These granules are dried and sieved to obtain granules of 177 to 1410:

(1) N-D-xyloside of p-amino

  sodium benzoate 45 parts (2) starch 15 parts (3) galactose 16 parts (4) crystalline cellulose 21 parts (5) polyvinyl alcohol 3 parts and (6) water used in processing to prepare the wet granule 30 parts

FORMULATION EXAMPLE 3

  Instead of using N-D-xyloside from p-

  sodium aminobenzoate in the formulation example 2, the sodium n-aminobenzoate NL-rhamnoside is used to prepare a granular formulation as in the formulation example 2. To 96 parts of the granular formulation thus prepared is added 4 parts of calcium stearate and mold the mixture

  compressed into tablets of 10 mm diameter.

FORMULATION EXAMPLE 4

  To 90 parts of granular formulation prepared in Formulation Example 2 are added crystalline cellulose parts and 3 parts calcium stearate, and the

  mixture prepared in tablets of 3 mm diameter.

  The tablets thus prepared are coated by means of a mixed suspension comprising gelatin in syrup and precipitated calcium carbonate for

  turn into sugar-coated tablets.

FORMULATION EXAMPLE 5

  It is intimately mixed by heating the constituents

  below and the mixture is introduced into

  bulbs and then sterilized for injection.

(1) N-L-rhamnoside of m-amino

  sodium benzoate 0.6 part (2) nonionic surfactant 2.4 parts and (3) aqueous solution of serum

physiological 97 parts.

EXAMPLE 1

  Effect of the substance of the invention on the metabolism of PGs of fixed arachidonic acid

in lymphocytes.

  After adjusting the lymphocytes taken from the spleen of a BALB / C mouse to a concentration of 1 × 10 7 cells / ml, 2 μCi of 3H-arachidonic acid is added and the mixture is incubated at a temperature of 370 ° C. for 90 minutes. . We wash three

  once the lymphocytes incubated with the culture medium.

  After again adjusting the cells to a concentration of 1 x 107 cells / ml, the culture is poured into four silicone test tubes at a rate of 2 ml / tube .- Two test tubes are used as controls, and in each of the two remaining test tubes, 500 μg of the substance of the invention is added, in this example sample No. 7, and the four test tubes are incubated at 370 ° C. for 60 minutes. After incubation, the test tubes are centrifuged at

  00C for 5 minutes at 1200 rpm.

  The granules of cells thus obtained are introduced into a flask containing 2 ml of the culture medium, and after adding 5 ml of petroleum ether, the flask is shaken and the ethereal layer is removed, the remaining aqueous layer being adjusted to pH. 3.5 with 0.5N aqueous hydrochloric acid solution. The acidic aqueous solution is extracted three times with 5 ml of ether each time, and the extract is dried.

  ethereal dry. The solid material is subjected to

  rification with a solution of diazomethane. The esterified reaction mixture was subjected to thin layer chromatography and developed with ethyl acetate: isooctane: acetic acid: water 90: 50: 20: 100 by volume. The identification of

  stains that appear on the chromatogram is ef-

  The following authentic samples of PGD 2, PGE 2, PGF 2 and 6-keto-PGF were removed by scraping the silica gel layer from the separated chromatogram and dissolving it in a liquid for counting in a scintillator. liquid. The modification of the PGs is determined from the variation of the counts. Table 5

gives the results obtained.

TABLE 5

  Prostaglandin modification in a culture of

cells in vitro.

| EoPGs aI PGF PGE PGF2-

PGF2 PGE2 -PGD-

Quantity) 6-lt-PG1

2) I

  500 +3) i I4) Notes: 1) added quantity of substance of the invention, sample No. 7 (g / ml), 2) no change is observed, 3) a slight change is observed,

  4) a clear change is observed.

  Similar results are obtained when samples Nos. 1-6 and 8-10 are used at 500 μg / ml. These results show that the various substances of the invention regulate the metabolism

  PGs even in the in vitro test.

EXAMPLE 2

  Effect of the substance of the invention on platelet aggregation: As is known today that platelet aggregation is caused mainly by PGG2, PGH2 and TXA2, all of which are PGs, the action of the substance of the invention on platelet aggregation was examined as follows:

  As an anticoagulant, a solution was used

  aqueous citrate for the determination of erythrocyte sedimentation; to a part of the citrate solution was added 9 parts of blood

  freshly collected human. The mixture was centrifuged

  It was run for 6 minutes at 400 x G and the supernatant was used for the preparation of platelet rich human plasma (PRP). The remainder was centrifuged again for 20 minutes at 700 x G and the supernatant was stored as platelet poor plasma (PPP). The change in PRP transmittance was measured in an aggregometer (PAP-3 type, manufactured by BIODATA, CO) to determine the degree of aggregation of platelets, aggregation being induced

by adding an agglutinating agent.

  The aglutinizing agents used were arachidonic acid (1.64 mmol), adenosine diphosphate (50 pmol), collagen (0.26 mg / ml), epinephrine (0.11 mmol), ristocetin (2.0 mg / ml) and thrombin (0.5 U / ml). Each agent was added to the PRP two minutes after the addition of one of the

  substances of the invention (samples 1 to 8).

  Figures 1 to 6 show the results

  obtained. We see that each of the substances of the invention

  This assay has an inhibitory effect against platelet aggregation caused by each of the aglutinating agents. This phenomenon shows that each of the substances of the invention regulates the production

  PGs, in particular those of PGG2, PGH2 and TXA2.

EXAMPLE 3

  Action of the substance of the invention on the

  cyclic adenosine monophosphate

  Cyclic AMP is known to be closely related to PGs in addition to its intracellular messenger function, and the relationship is closest to PGD, PGE, and PGI. Accordingly, in the present example, the influence of the substance of the invention on cyclic AMP in the platelet cell was examined as follows: PRP (Platelet Rich Plasma) was prepared by method as above and was centrifuged for 700 x G for 20 minutes to obtain a supernatant liquid called PPP (platelet poor plasma). The precipitate was resuspended in 1/3 times by volume of PPP to prepare a PRP

concentrated three times.

  In the concentrated PRP thus prepared, each aqueous solution of the substances of the invention has a pre-established concentration (in this case, samples

  Nos. 4, 7 and 8) and an aqueous solution of physiological serum

  logic, and the mixture was incubated at the same temperature

  room temperature for 5 minutes. Then the boiling, homogenization and centrifugation operations were carried out in that order, and 50 μl of the supernatant was used to assay the cyclic Δ4P. The measurement was made by Gilman's method. As a positive control, PGE1 was used. The results are given in Figure 7. As shown in Figure 7,

  the substance of the invention has an action of

  cation of cellular intra-platelet cyclic AMP

  silence, and this fact suggests that the substance of

  the invention affects the metabolism of PGs.

EXAMPLE 4

  Action of the substance of the invention on the malondialdehyde (MDA) platelet count:

  PRP collected from human blood has been centri-

  leaked and washed to form "washed platelets".

  After adding one of the substances of the invention, (Samples Nos. 4, 7 and 8) to the washed platelets, Caionophore A23187 was further added thereto and the mixture was incubated for 5 minutes at 37 ° C.

  thiobarbic acid was added to the incubated mixture.

  turkey, as a color developing agent and the mixture was extracted with a mixture of methanol and butanol. Absorption at 535 nm was read colorimetrically to determine the amount of malondialdehyde (MDA). Figure 8 gives the results obtained. The substance of the invention suppresses the production of MDA. This fact suggests that he too

  the participation of the substance of the invention in the metabolism

the PGs.

EXAMPLE 5

  Effect of indomethacin on the hypo-

  Tensive of the substance of the invention: It is known that PGs are formed from arachidonic acid and that cyclooxygenase participates in the conversion of arachidonic acid to PGG2, that indomethacin inactivates cyclooxygenase above and further that the metabolism of PGs is completely

  arrested by the administration of indomethacin.

  As a result, the change in blood pressure of spontaneously hypertensive male rats (SHR) 30 to 40 weeks after birth was compared in both cases (1) where only the present substance is administered at 100 mg / kg and 2) Indomethacin is administered twice at 2.5 mg / kg / time before and after administration of the substance of the invention. The substance of the invention, Sample No. 7, was dissolved in distilled water or

  dispersed in a 2% aqueous solution of carboxy

  methylcellulose, then forcibly administered orally. Indomethacin was dispersed in a 2% aqueous solution of carboxymethylcellulose, and the dispersion was also forcibly administered orally one hour before and after administration of the substance.

of the invention.

  The results are given in Figure 9.

  The hypotensive effect of the substance of the invention disappears when indomethacin is administered before and after administration of the substance of the invention. In view of the fact that indomethacin is an inhibitor of prostaglandin metabolism, the efficacy of the substance of the invention for the reduction of blood pressure can be attributed to

  to its close relationship with prostaglandins.

EXAMPLE 6

  Action of the substance of the invention on the cyclic AMP level in the tumor cell of sarcoma 180: The substance of the invention was added to 100 μl of a tumor of the ascitic type taken from the abdominal cavity of carrier mice sarcoma 180 at a pre-determined concentration, in this case Sample No. 7, and the mixture was

  incubate at room temperature for 5 minutes.

  After incubation, the culture was boiled, homogenized and centrifuged to obtain a supernatant liquid. The supernatant liquid thus obtained was subjected to the determination of cyclic AMP by the Gilman method. The results are given in

  Figure 10. It has been found that the substance of the invention

  The effect was to increase cyclic AMP levels in sarcoma 180 tumor cells. This fact also suggests that

  substance of the invention to the metabolism of PGs.

EXAMPLE 7

  Influence of the substance of the invention on

  PGI2 productivity of 3T3 fibroblasts.

  The influence of the substance of the invention on the PGI2 productivity of mouse 3T3 fibroblasts was examined. As a witness, we used a

  culture in which precursor arachidonic acid

  PGs, was added to 3T3 mouse fibroblast culture medium, and the mixture was incubated for 5 minutes at 370C to produce

PGI2.

  In addition, 30 mmol was added. of each of the substances of the invention (Sample Nos. 1, 3, 4 and 7-11) to 4 ml of 3T3 cell culture medium, and the mixture was incubated for 2 minutes at 370C. After adding arachidonic acid to the incubated culture, it was incubated again as for the control to produce PGI2. This culture

was called the treated group.

  After obtaining the respective supernatant liquids from the cultures, the temn group and the treated group, their PGI2 productivity was compared using the suppressive action on the aggregation of

  platelets caused by the addition of 2.43 mmol.

  arachidonic acid as an indicator.

  Figure II shows the results obtained; it can be seen that although the suppression of platelet aggregation is noted to a certain extent for the control after 5 minutes after the addition of arachidonic acid as precursor of the PGs, there is a remarkable suppression in the treated group, indicating the increase in productivity

  PGI2 by addition of the substance of the invention.

EXAMPLE 8

  Effect of the substance of the invention on the metabolism of cyclic AMP and cyclic guanosine monophosphate in a cancer-bearing mouse Ehrlich cancer cells were transplanted into the abdominal cavity of ICR / JCL female mice five weeks later their birth

  at a rate of 1 x 106 cells / animal, and while feeding

  both the mice, the substance was intraperitoneally administered

  In the present invention, sample No. 7, mice daily from the day following transplantation, for five days, at a rate of 200 mg / kg / day. On the sixth day, the mice were sacrificed with ether to collect on each 5 ml of ascites. After adding tetrasodium ethylenediaminetetraacetate to the ascites thus collected, the mixture was centrifuged for 10 min at 4 ° C and 1500 rpm to separate the supernatants and the cancer cells. The supernatant liquids were again

  centrifuged at 4 ° C. and 3000 rpm for 15 minutes.

  The separate cancer cells were washed with

  a solution from Hank, then 5 times centrifugally

  repeated at 800 rpm for 5 min each time to remove impurities such as erythrocytes, etc. After homogenizing 1 x 10 8 cancer cells in 3 ml of a 6% aqueous solution of trichloroacetic acid at 0 ° C, the homogenate was centrifuged for 20 minutes at 3000 x G to obtain a liquid extract. 10% by weight of 1 N aqueous hydrochloric acid solution was added to the liquid extract and then extracted five times with each

  2 volumes of ether to remove trichloroacid

  tetraacetic. After complete removal of the ether by heating the residual liquid in a water bath at 80 ° C.,

the residue was lyophilized.

  To the ascites supernatant was added an equal amount of a 10% aqueous solution of trichloroacetic acid and, after standing for 15 minutes at 0 ° C, was centrifuged for 20 minutes. at 3000 x G to obtain a supernatant liquid. To the supernatant liquid thus obtained, was added 0.1 ml of a 1N aqueous solution of hydrochloric acid, and after removing the trichloroacetic acid by two volumes of ether, the ether was completely removed in a water bath. 80 C, and we have

lyophilized the residue.

  Freeze-dried samples from the cancer cells and the ascites supernatant were respectively dissolved in 1 ml of aqueous buffer solution. PH 4.0 containing 50 miol of acetic acid and subjected to

  the radioimmunoassay using anti-

  body anti-cyclic AMP and anti-morno-

  cyclic guanosine phosphate (GMP) to determine cyclic AMP and cyclic GMP respectively in cancer cells and ascites supernatants. Table 5A gives the results obtained. The increase of the cyclic AMP and cyclic GMP contents in the cancer cells was noted, the cancer cells from a rat to which the substance of the invention had been administered. This fact shows the influence of the substance of the invention on the in vivo metabolism of cyclic AMP and cyclic GMP in cancer cells from cancer cells transplanted into the body.

of the mouse.

TABLE 5A

  Cyclic AMP and Cyclic GMP Rates Unit: pM

SR 1367 JA MDT

Cyclic AMP for cyclic GMP for

  Group 108 cells can- 3.08 cells can-

  Figure 19.5 0.59 Substance of 21.2 0.72 antigen

EXAMPLE 9

  Effect of the substance of the invention on the amount of PGs in a culture medium in which cells of human leukemia are cultured: 10 ml each of a culture medium prepared by adding 10% of serum bovine fetus and one of the substance of the invention, Sample No. 7, at a concentration of 50, 500 or 5000 μg / ml to an Eagle culture medium, and placed in a 75 cm polystyrene bottle from the background surface, 105 cultured cells of human dinucleate leukemia strain J-111 are innoculated and cultured at 370 ° C. in a mixed atmosphere of 5% by volume of gaseous carbon monoxide and 95% by volume of air during 7 days. During the culture, the culture medium is replaced on the second day and the fourth day with fresh media, and each of the spent media is subjected to centrifugation as soon as possible at a temperature of

  C at 1500 rpm to obtain each supernatant liquid

  from the culture medium. The PGE and PGF2 content in each supernatant fluid is determined using a radioimmunoassay kit for 3H-Prostaglandin E and a H-Prostaglandin F radioimmunoassay kit 3.

by U.S.A. Clinical Assay Co.).

  The results are given in Figs. 12 and 13. As shown in Fig. 12, the cumulative amount of PGF 2 has increased over time and Fig. 13 shows that the cumulative amount of PGE decreases with time, generally. . However, the speed

  The increase in PGF2 has decreased when the concentration of

  the substance of the invention increases, and the speed of

  The decrease in EMP increases when the concentration of

  tion of the substance of the invention increases. In any case, the participation of the substance of the invention

  to the metabolism of PGs is shown in Figures 12 and 13.

EXAMPLE 10

  Effect of the substance of the invention on the amount of PGs in a culture medium in which human uterine cancer cells are cultured: To 10 ml of a culture medium prepared by adding 10% fetal bovine serum and a of

  this substance, sample No 7, at a concentration

  of 0 (control), 10, 100, 500, 1000, or 5000 μg / ml to Eagle culture medium and placed in a polystyrene flask whose bottom has a cm surface, HeLa cultured cells are innoculated. S3 of human uterine cancer at 5 x 105 cells / vial and cultured at 370C for 2 days in a mixed atmosphere of 5% by volume of carbon monoxide gas and 95% by volume of air. When the incubation is complete, the entire culture medium is centrifuged at a temperature of 40 ° C. at 1500 rpm to obtain a supernatant liquid of the culture medium. The PGE content of the supernatant

  determined using the radio test kit

  Immunological Evaluation of 3H-Prostaglandin E. Results

are given in Table 6.

  As shown in Table 6, the quantity

  of PGE in the culture medium (without cancer cells)

  reuses) is reduced by the addition of the substance of the invention. This fact shows the effect of the substance

  of the invention on the metabolism of PGs.

TABLE 6

  PGE content in the culture medium (without cells)

  Concentration of the Concentration of the Variation of the Con

  substance of the PGE after culture centering of the PGE vention (lg / ml) (pg / ml) during the culture (pg / nl)

0 312 +120

40 -152

100 80 -112

500 48 -144

1000.25 -167

5000 25 -167

EXAMPLE 11

  Effect of the substance of the invention on PGE in cancer cells and in ascites of mice transplanted with Ehrlich cancer cells: To groups of ICR-JCL female mice at five weeks of their birth, intraperitoneal transplants were transplanted intraperitoneally. Erwinich cancer cells at 1 x 10 6 cells / animal and one of the present substances, Sample No. 7, was administered intraperitoneally to the mice daily, from the day following transplantation for five days, at a dose of 200 mg / day. kg / day. The mice are sacrificed with ether on the sixth day after transplantation,

  and we collect the ascites. After addition of ethylene

  Tetrasodium diaminetetraacetate with 1% by weight aspirin to ascites, centrifuged for 10 min at 1500 rpm and at a temperature of 4 C for

  separate the cancer cells from the supernatant fluid.

  The supernatant liquid is again centrifuged at 4 ° C. and 3000 rpm for 15 minutes. The separated cancer cells are washed with Hank's solution and centrifuged five times at 800 rpm for 5 minutes each time to remove impurities such as erythrocytes and the like. The cancer cells thus purified, at a number of 1 × 10 8, are homogenized at 0 ° C. after addition of 7 ml of methanol, and the homogenate is filtered on filter paper. After addition of 14 ml of chloroform to the filtrate and mixture, the mixture is left

* Mix for 30 minutes at a temperature of 40C.

  Then the precipitated protein is removed by suction

  filtration, and the filtrate thus obtained is evaporated to dryness in a rotary evaporator. The dried solid material is placed in a separatory funnel, with chloroform, methanol and a dilute aqueous solution of hydrochloric acid at pH 2, and after thorough mixing, is dissolved in the lower aqueous layer of a solution. 2 mil solution. The PGE content of the solution and supernatant fluid of ascites is determined using a radioimmunoassay kit for 3H-Prostaglandin E. Table 7 gives the results obtained. In the animal to which cancer cells were transplanted and administered the substance of the invention, the PGE is contained in a larger amount in the cancer cells and less in the ascites supernatant fluid than the animals to which transplanted the same cancer cells, but not administered the substance of the invention. This fact strongly suggests a relationship between the administration of this substance and

the metabolism of PGs.

TABLE 7

  PGE content in cancer cells and ascites (supernatant fluid)

EXAMPLE 12

  Effect of the substance of the invention on the amount of PGE in Erhlich cancer cells and on the proliferation of tumors

  Erhlich cancer cells are trans-

  planted subcutaneously to groups of female C57BL / 6 mice nine weeks after birth at 1 x 106 and. while feeding the mice, the substance, sample No. 7, is administered intraperitoneally to the mice every day after the day following the transplant, at a rate of 100 mg / kg. On the fourteenth day after transplantation, one sacrifices

  the mice with ether to remove the tissue. tumor.

  After homogenizing the finely cut tumor tissue with scissors, adding 7 ml of methanol per gram of tumor tissue at a temperature of 0 ° C, the homogenate is filtered with filter paper to provide a filtrate. After adding two volumes of chloroform to the filtrate, the mixture is thoroughly mixed and left for 30 minutes at 40C. Then we submit the mixture

  cooled to the same operations as in example 11 to determine

  undermine the PGE content in the tumor tissue. Table 8 shows the results obtained. The PGE content in the tumor tissue is higher in the animal to which cancer cells have been transplanted and the PGE content in the PGE content is administered.

108 cells the supernatant fluid

  Group (ng) ascites (ng) Control 2.36 0.26 Substance of 3.06 0, 12 the invention only substance in the control to which the same cancer cells were transplanted but not administered the substance of the invention. This fact gives information on the participation of the substance of the invention in the metabolism of PGs.

TABLE 8

PGE content in tumor tissue

EXAMPLE 13

  Repetition of the experience of Example 12

  in slightly different conditions.

  Erb-cancer cells are transplanted

  lich subcutaneously to groups of C57BL / 6 female mice eight weeks after birth and, while feeding the mice, the substance of the invention, Sample No. 7, is administered orally,

  to each mouse, each day, at a rate of 1 g / kg / day.

  The seventh or fourteenth day after the transplan-

  The mice are sacrificed with ether to remove the tumor. The tumor is treated according to the same procedure as in Example 11 to determine its PGE content. Table 9 gives the results obtained. The concentration of PGE in the tumor is higher in the animal to which cells have been transplanted and administered the substance of the invention than in the control to which the same cells were transplanted but not administered the substance of the invention. In addition, it is observed that the PGE content decreases in both groups over time, and that the rate of decrease of tissue weight PGE content in tumor group tumor tissue (g) (rig / g) Control 1.30 1 , 77 Having received the compound of 0.75 3.98 the invention

  is lower in the group that received the transplan-

  tion and administration of the substance of the invention than in the control group

  but not the administration of the substance of the invention.

  These facts show that the substance of the invention

  participates in the metabolism of PGE.

TABLE 9

PGE content in the tumor

EXAMPLE 14

  Effect of the substance of the invention on the PGE content of the tumor of adenocarcinoma 755 and its proliferation Subcutaneous transplantation of adenocarcinoma 755 tumor cells to groups of female C57BL / 6 mice at 8 days of age. birth and, while feeding the mice, are given orally every day starting the following day

  transplantation, the substance of the invention,

  No. 7, at a rate of 1 g / kg / day. The mice are sacrificed with ether at the sixth or fourteenth

  day after transplantation to root out the tumor.

  The tumor thus eradicated is treated according to the same procedure as in Example II to determine its PGE content. Table 10 gives the results obtained. As can be seen, the PGE content in the tumor decreases over time in the two groups, one of which has received the transplantation and the date of delivery. - in the transplant transplantation slug (g) (ng / g) Control 7th day 0.09 2.59 14th day 0.84 1.16 Having received the 7th day 0.08 2.80 substance of the invention 14th day 0 , 67 1.65

  and the other has received the transplant but not the

  therefore, the PGE content is higher in the group administered the compound of the invention than in the control. These facts show the participation of the compound of the in-

  the metabolism of prostaglandins.

TABLE 10

PGE content of the tumor

ETXIPLE 15:

  Effect of the substance of the invention on the migration of a dye from the injection region to the transplanted tumor: On the back of a donryu rat, a piece of 5 mm Sato lung cancer is transplanted by

subcutaneous route.

  In the axillary part of an ICR mouse, sarcoma 180 cells were transplanted by the subcutaneous route. While feeding the two animals, the substance of the invention, sample No. 7, was orally injected once with 1000 mg / kg at sixteenth day after transplantation, and a 2% aqueous solution of Lissamine Green (dye manufactured by Imperial Chem Ind. Co.) was injected into the tail vein of the

  1 ml for unrat and 0.5 ml for a mouse.

  Expiration Date PGE Weight in Group pation after tumor tumor transplantation (g) (ng / g) Control 7th day 1.34 0.67 14th day 4.01 0.34 Having received 7th day 1 , 04 2.83 the substance of the invention 14th day 3.10 1.15 tion

  One hour after the injection, the animal is sacrificed

  badly to extirpate the proliferating tumor. The rat tumor is finely cut and homogenized with a 50% aqueous solution of ethanol. After diluting the homogenate by adding the solution to a total volume of 50 ml, the liquid is centrifuged for 15 minutes at 1000 rpm to separate the supernatant liquid. The dye content of the supernatant liquid itself or after dilution by spectrophotometry is determined using its maximum absorption of 630 nm. With regard to the tumor of the mouse, it is cut out and the presence of dye is observed

  with the naked eye on the cross section of the tumor.

  The amount of dye that has sedimented in the lung cancer of Sato per gram of cancer is

shown in Table 11.

TABLE 11

  Group Amount of dye (pg / g of tumor)! Witness 30.0 Having Received the Substance of 51.0 the Invention

  The visual examination shows the remarkable

  dye in the central part of the tumor in the case of sarcoma 180 in the mouse

  received the administration of the substance of the invention.

  These results show that the substance of the invention facilitates the access of an anticancer drug to a tumor and suggests the participation of the substance.

  of the invention to the metabolism of PGE.

EXAMPLE 16

  Effect of the substance of the invention to prevent the metastasis of a transplanted tumor: MH-134 tumor cells were transplanted to a C3H / He mouse by its tail vein at 2 x 10 6 cells / animal. The substance of the invention, Sample No. 7, is orally administered to the mouse, respectively, 6, 3 and 1 hour before and 1, 3 and 6 hours after transplantation, at a rate of 1000 mg / ml. kg. The mouse is sacrificed on the fourteenth day after transplantation to extirpate the lungs

  to examine the number of metastatic lesions.

  Table 2 gives the rate of positive metastases

  and the number of metastatic bonds.

TABLE 12

  Metastasis rate Number of metastatic positive group linkages (%)

Having received ad-

  Administration 6 hours before 10/10 (100%) 2.3 3 hours before 8/9 (88.9%) 1.9 1 hour before 7/9 (77.8%) 2.2 1 hour after 7/10 (70%) 2.6 3 hours after 9/10 (90%) 3.0 6 hours after 10/10 (100%) 3.5 Not having received tH 6/6 (100%) 4, Administration 6/6 (100%) 4.5 -With many information is available regarding the involvement of PGs, particularly PGD2 in the metastasis of a transplanted tumor, the above results are considered to be suggestive. an inhibitory effect of the substance of the invention

  on the metastasis of a tumor implanted by

mediator of PGs.

EXAMPLE 17

  Variation of the level of PGs in the blood of the animal to which the substance of the invention has been administered: Methylcholanthrene-induced sarcoma cells are transplanted on the back of a group of spontaneously hypertensive rats (SHR) by injection. subcutaneous

  at a rate of 1 x 106 cells / animal and, while at the same time

  In rats, the substance of the invention, Sample No. 7, is administered daily intraperitoneally for 6 consecutive days from the day following transplantation, at a dose of 500 mg / kg / day. Two weeks after the end of the administration, whole blood is collected to obtain a fraction

  of plasma. The ethereal extract is separated from the plasma by

  thin layer chromatography and after trans-

  The extract was formed into a methyloxymésilyl derivative and the variation of 6-keto-PGF1a in the extract was examined by gas chromatography and mass spectroscopy. Figure 14 gives the results

obtained.

  It is seen that, although in general the level of 6-keto-PGF1 in the blood of the rat infected with cancer is higher than that in the blood of a rat in the normal state, the rate in the rat bearing a cancer administered with the substance of the invention is almost the same as in the normal rat. It is presumed that the substance of the invention has reduced the

increased to its normal value.

EXAMPLE 18

  Effect of the substance of the invention on the proliferation of cancer and on the amount of PGE in cancer cells: By the same method as in Example 12, the anti-cancer action against cancer was studied. Transplanted Erhlich in the mouse C57BL / 6, and the amount of PGE in the proliferating cancer cells in the body of the mouse, in the case of the administration of each of the compounds of the invention, samples n 1, 2, 3, 4, 5, 8, 9, 10 and 11, intraperitoneally every two days from the day after transplantation

  a total of six times at the dose of 100 mg / kg.

  The results of the determination of the weight of the cancer extirpated from the sacrificed mouse fourteen days after the transplantation of cancer cells, 1 x 10 per animal, and the PGE content of the extirpated cancer are shown in Table 13 below:

TABLE 13

  Cancer Weight (g) and Cancer PGE Content (ng / g) Cancer Weight PGE Content of Compound (g) Cancer (ng / g) Control1) 1.30 1.77 Sample n 1 0.63 4, 10

2 0.81 3.88

3 0.74 4.23

4.0.56 4.19

0.70 3.20

0.65 3.66

72) 0.75 3.98

8 0.72 4.03

9 0.80 4.32

0.70 4.14

11 0.69 4.20

  Notes: 1) animals that have received the transplant,

but not the administration.

2) Results of Example 12.

  As shown in the table, although there are some differences between the compounds, they all effectively inhibit cancer growth and all effectively raise the PGE content of the cancer cells.

EXAMPLE 19

  Effect of Aspirin on the Hypotensive Action of the Substance of the Invention: It is known that PGs are formed from arachidonic acid and that conversion of arachidonic acid to PGG2 passes through cyclooxygenase. It is also known that aspirin inactivates cyclooxygenase, and that the metabolism of PGs is completely stopped by aspirin administration.

to a mammal.

  Therefore, the following experiment was performed: at groups of male 30- to 40-week-old SHR rats, (a) one of the substances of the invention was administered orally at 100 mg / kg. as a solution in distilled water or an aqueous suspension in a 2% solution of carboxymethylcellulose, or (b) forcibly aspirated aspirin, one hour before and one hour after the administration of the substance of the invention at 200 mg / kg. The blood pressure of the rats thus treated was measured as a function of time. The results are given in

Table 14.

  As Table 14 shows, by adminis-

  Inhibition of the metabolism of PGs, aspirin, the hypotensive effect of

  the invention (samples 4, 7 and 8) which appear

  knows in the group (a) did not appear in the

  group (b). The results show that the hypo-

  tensor of the substance of the invention is due to the PGs.

TABLE 14

Hypotensive effect inhibited by

the aspirine-

  Notes: 1) 6 hours after the administration of the

of the invention.

  2) Only distilled water was administered at the same time as administration of

  aspirin and the substance of the invention.

EXAMPLE 20

  Effect of the substance of the invention on the

  rate of PGs in the blood of a rat spontaneously hyper-

  Tensif (SHR): After forcing the substance of the invention (samples 4, 7 and 8) daily for 14 consecutive days at a rate of 100 mg / kg / day orally to groups of male rats SHR, their whole blood was collected on the fifteenth day to obtain the plasma fraction. The ether extract of the plasma fraction was separated and isolated by thin layer chromatography, and after

  transformed the isolated fraction into a methyloxymesi-

  lysed, the alteration of 6-keto-PGFlc by gas chromatography and spectroscopy was studied.

  massive. The results are given in Table 15.

  In the group to which the substance of the invention was administered, the 6-keto-PGF ia content was higher. Modification of the blood pressure (mmHg) i Compound Case (a) Case (b) Witness OO Sample N 4 20 - 5

N 7 - 20 O

N 8 - 15 O

  and the blood pressure is lower than in the control.

  This parallelism of the effects leads to suppose that

  the effect of the present invention passes through the

Prostaglandins.

TABLE 15

  Blood pressure and PGs content of the blood collected 14 days after the administration of the substance of the invention (samples N 4, 7 and 8)

EXAMPLE 21

  Effect of the substance of the invention on PGI2 production in a rat with diabetes mellitus: Groups were used for these experiments

  Male Sprague-Dowley rats with a body weight of

  viron 200 g. In some groups of rats streptozotocin was administered intravenously at 80 mg / kg one to three months before the start of the present experiment so that they were in a state of diabetes mellitus. . Rats of the same strain, same sex, and age were used as controls

  and who have not received streptozotocin administration.

  The substances of the invention (samples N 4, 7 and 8) were forcibly administered to groups of rats with artificial diabetes mellitus orally, at 100 mg / kg in the form of

  aqueous solution in distilled water.

  TESTIMUM Sample-Sample-Sample-Sample

  lon n 4 1Ion 7 lon n 8, t Blood pressureJ Blood pressure 195 155 160 150 I (imffg) Blood content in 6-keto-PGF 1-O 3 7 6.5 7 (ng / ml) Six hours after the administration of the substance of the invention, a sample of blood was taken from the rat under anesthesia with ether, at a rate of 0.5 ml / animal, and the sugar content of the blood was measured by the enzymatic method. .

  The amount of prostitution was then measured

  I2 glandin produced in the arterial tissue as follows: The thoracic aorta was rapidly removed from the rat, washed with Krebs buffer solution at a temperature of 40C and cut out

  in thin rings of 1 to 2 mg. Then, in 200 micro-

  of Krebs buffer solution, mg of the thus cut aorta was introduced and incubated.

  mixing for 3 minutes at 220C.

  The amount of prostaglandin 12 (PGI 2) produced by the cut aorta was determined in the supernatant fluid (1 to 10 microliters) of the above incubated culture in the same manner as in Example 2, using as an index the suppressive action of platelet aggregation, but using as agglutinant diphosphate

  adenosine. After preparing a yardstandard

  Using an authentic PGI2 sample, the PGI2 content of the above supernatant was determined from the calibration curve in ng / mg wet weight of the arterial tissue. Table 16

gives the results obtained.

TABLE 16

  Quantity of PGI2 and Blood Sugar Although PGI2 production in rat aortic tissue artificially induced in a state of diabetes mellitus is lower than that of normal rat, administration of the substance of the invention to a rat brought artificially into a state of diabetes mellitus increases the ability of the tissue to produce PGI2

  while reducing the blood sugar level.

EXAMPLE 22

  Another example showing the effect of the substance of the invention on the production of PGI2: After extracting the cervical aorta from an SD rat under anesthesia, it was washed with a Krebs-Linger bicarbonate buffer solution and it was cut into thin rings, and the chopped pieces were stored in the above buffer solution at the rate of 1 mg of wet cut pieces per 0.5 ml to produce

the PGI2.

  PGI2 Sugar in Group of rats (ng / mg wet tissue blood) (mg / 100 ml) Normal rat 0.26 0.06 76.7 - 5.0 Rats with artificial diabetes mellitus Not treated with + substance of 0.07 - 0.01 412.1 + 15, 1 treatment treated with e-0.19 0.04 133.6 + 8.3 sample 4 9 0.04

Treated by the

  Sample No. 7 0.22 + 0.02 165.0 + 11.0 Treated with e-0.18 + 0.04 110, 1 + 5.5 Sample No. 8 At 500 microliters of the mixture thus prepared, 50 microliters of a physiological saline solution containing 2% by weight of the substance of the invention (each of samples Nos. 4, 7 and 8) are added and, as PGI 2 production generally reaches a maximum after After 10 minutes of storage, a supernatant liquid (2.5 μl) of the mixture was prepared 10 minutes after the addition, and then the amount of PGI 2 in the supernatant was determined using the suppressive action of platelet aggregation. as an index and using ADP as an agglutinating agent, the control being a simple saline solution. Figure 15 gives the results obtained. The addition of the substance of the invention increases the production of PGI2 by

  the cut pieces of the cervical aorta.

  In addition, although various studies have been conducted on the cause of arteriosclerosis, agglutination of aggregated pieces of the vessel wall is indicated as a cause of particular importance. In other words, preventing the agglutination of platelets on

  the vessel wall is the first treatment.

  It is known that PGI2 has a pronounced inhibitory effect on platelet agglutination, so that the substance of the invention which increases the possibility of producing PGI2 on the aortic wall has a treatment effect and a preventive effect.

arteriosclerosis.

EXAMPLE 23

  Elevating effect of the substance of the invention on the level of PGE1 in the mammalian organism: (1) In the case of artificial foot edema A three groups of male Sprague-Dowley rats out of a total of six groups, forcibly administered

  orally, the substances of the invention,

  samples Nos. 4, 7 and 8, at the rate of 1000 mg / kg, the other three groups of the same species of rats

receiving nothing at this time.

  Sixty minutes after administration, 0.1 ml of an aqueous solution of carcinoin (1 mg) and PGE1 (0.1 μg) were injected into the hind paws of the four groups of rats including the rats.

  three groups having received the substance of the invention.

  To one of the two remaining groups not treated with the substance of the invention, 0.1 ml of an aqueous solution containing only carrageenan was injected.

  1%, and to the other group that did not receive the

  of the invention, an aqueous solution containing no

than PGE1.

  The volume of the swollen foot of the groups of rats thus treated was measured by the water displacement method, and the results are given in Table 17, expressed by the increase in the percentage of the volume of the foot relative to the volume.

of the foot before treatment.

  (2) In the case of skin stimulation To the three groups of male Sprague-Dowley rats out of the total of 6 groups, the substances of the invention, samples Nos. 4, 7 and 8, were administered by force. 1000 mg / kg, the other three groups receiving nothing at this time. Sixty minutes after administration, a mixture of PGE (2.5 x 10 11 mol / animal) and histamine (2 x 10-8 mol / animal) was injected intracutaneously under mild anesthesia. ether in the shaved skin of an abdominal portion of four groups of rats comprising three groups having received the substance of the invention. To a group of the two groups remaining intact, only histanine was injected at 2 x 10-8 mol / animal and to the other group

  only PGE, at a rate of 2, 5 x 10 11 mol / animal.

  Immediately after the above injection, Evans blue was injected as an aqueous solution in physiological saline solution, into the lateral vein of the tail of all experimental rats at 2 ml / kg corresponding to mg of Evans blue / kg. All animals were sacrificed 30 minutes after dye injection, and the extent of dye leakage was studied spectrophotometrically by the Harada method.

  et al. using as an index the absorbance at 620 nm.

  The results are given in Table 18.

TABLE 17

  Effect of the substance of the invention on carrageenan induced swelling

Percentage increase

  Groups of rats treated with foot volume

  Groups 60 minutes after administration

  by carrageenan Carragenin 37.0

PGE19,3

  Carragenin + PGE1 66.2 Sample n 4 + 32.5 Carrageenan + PGE1 Sample n 7 + 28.2 Carrageenan + PGE1 Sample n 8 + 29.7 Carrageenan + PGE1

TABLE 18

  Effect of the substance of the invention on dye leakage. The results in Table 17 indicate that carrageenan-mediated inflammation enhanced by PGE1 is inhibited by

  the action of the substance of the invention.

  The results in Table 18 indicate that dye leakage due to histamine and enhanced

PGE1 is inhibited.

EXAMPLE 24

  Diuretic effect of the substance of the invention

  depending on the level of 6-keto-PGFla in the blood of rats.

  Groups of spontaneously hypertensive female rats (SHR) kept fast for 12 hours and deprived of water for 2 hours were treated with

  forced oral administration of the substance of the invention

  (each of samples 4, 7 and 8), with the respective values of 1 and 5 g / kg dissolved in 8 ml (for

  by weight) of an aqueous solution of physiological serum

  the control group receiving only the aqueous solution of physiological saline at a rate of

8 ml / 200 g by weight.

  Group of rats treated with 620 Histamine Absorbance 0.192

PGE1 | 0.066

  Histamine + PGE10,259 Sample n 4 + histamine + 0,155

PGE 0.155

  PGE1 Sample n 7 + histamine + 0.162 PGE1 Sample n 8 + histamine +0.6, 16 The amount of urine excreted by the rats was measured every hour from one hour after administration, six consecutive times, and the percentages of the volume of the urine to the volume of saline solution containing the substance of the invention initially administered (8 ml, per 200 g by weight) are given in

table 19.

  Then, six hours after the administration, all the animals were sacrificed to collect

  their whole blood to obtain its plasma fraction.

  The ether extract of the plasma fraction was separated by thin layer chromatography and, after transforming the thus separated substances into methyloxymésilyl derivatives, the variation of 6-keto-PGF1 was studied. by gas chromatography-mass spectrography. The results are

  also given in Table 19.

  As shown in Table 19, the amount of urine, after conversion to the normal recovery value, was greater in the rats that received the substance of the invention than in the controls, and a dose-response relationship was found about the amount of urine among the rats that received the substance of the invention. In other words, a diuretic effect has been observed for the substance

of the invention.

  In addition, the level of 6-keto-PGF1a in the blood of the rats which received the substance of the invention is a little higher than for the control, although the difference is very small. Although it is natural that the increase of 6-keto-PGF1a, which is the metabolite of PGI2, results in an increase in the PGI2 level, as the diuretic function is recognized as one of the physiological functions of PGI2, it is is to assume that the substance of the invention must

  its diuretic effect to the regulation of PGI2.

TABLE 19

  Quantity of urine and 6-keto-PGFlo in the blood Volume of urine1)% Rate of 6-keto-PGF1a groups 1 2 3 4 5 6 in the blood hours (ng / ml) Control 2 9 22 26 3.0 Substance of the invention Sample N 4 g / kg 7 40 50 52 78 91 3.5 1 g / kg 6 40 46 50 60 63 3.3 Sample N 7 g / kg 5 50 66 80 92100 3 , 9 1 g / kg 4 38 45 48 63 72 3.5 Sample N 8 g / kg 5 43 60 70 77 93 3.7 1 g / kg 5 39 47 50 65 68 3.2 _ Note: 1) Percentage volume of accumulated urine at the time of determination based on the volume of saline solution

  containing the substance of the invention

istered.

EXAMPLE 25

  Effects of the substance of the invention on the threshold of pain caused by repeated injections of

PGE2:

  To a group of Wistar male rats with a body weight of 230 to 250 g was injected daily for 2 weeks, in both legs, 2 μg of

  PGE2 dissolved in 0.1 ml of a sterile aqueous solution

  0.9% sodium chloride. At the sixteenth day, the substance of the invention (each of samples 4, 7 and 8) was forcibly administered by

1000 mg / kg.

  The "pain threshold" was measured one hour after administration of the substance of the invention. The term "pain threshold" refers to

  the minimum pressure applied to the rat paw

  voicing anxiety in the rat. The results are

given in Figure 16.

  As shown in FIG. 16, the reduction of the "pain threshold" caused by the injection of PGE 2 is reduced to almost zero by administration of the substance of the invention. In other words,

  the substance of the invention has an analgesic effect.

EXAMPLE 26

  Effect of preventing the occurrence of stress ulceration in the stomach of rats by the substance of the invention: Three groups of male Donryu rats with a body weight of 220 to 250 g were administered orally after 23 days. , 5 hours of fasting, the substance of the invention (each of the samples No. 4, 7 and 8), at a rate of 1 g / kg dissolved in 8 ml (for 200 g of

  body weight) of saline solution.

  These three groups of rats and another group of rats also fasted for 23.5 hours and then received only the saline solution were placed in a stress cage, and immersed in water at a temperature of 230C until 'to the

* chest to cause stress in rats.

  After seven hours of immersion, the rats are removed from the water and are immediately sacrificed to collect their whole blood, and their stomachs are extracted. The plasma fraction prepared from the blood is extracted with ether and the ethereal extract is separated by thin layer chromatography. The separated extract after transformation into the respective methyloxymésilyl derivatives, is subjected to a gas chromatography-mass spectrography to determine its 6-keto-PGF 1a content. A 1% aqueous solution of formaldehyde is injected into the extracted stomach and, ten minutes after the injection, the stomach is cut to open on the side of the largest curvature to examine the mucous membrane of the gastric body. . The greatest extent of each erosion of the mucous membrane of the rat, possibly present, is measured under a microscope, and its sum expressed in millimeters is taken as the ulceration coefficient of the rat. The results are

given in Table 20.

  As shown in Table 20, the substance of the invention effectively suppresses the onset of stress ulcer, and the amount of 6-keto-PGF1a in plasma is higher in rats having

  received the substance of the invention only in the control.

  Since the increase in 6-keto- PGF1i, which is a product of PGI2 metabolism, means an increase in PGI2, and PGI2 is recognized as active in suppressing secretion of gastric juice and in suppressing ulcer formation, the substance of the invention is considered to be present

  an anti-ulcer effect by regulating the level of PGI2.

TABLE 20

  Stress ulcer and 6-keto-PGFla level: Note: 1) Sum of the largest extents of the mucosal membrane erosions of the treated rat groups divided by that

of the witness, multiplied by 100.

EXAMPLE 27

  Effect of the substance of the invention on

  pyrexia caused by a pyrogenic bacterium.

  The experiments were carried out on cats weighing 3 to 3.5 kg. In aseptic conditions,

  sodium pentobarbitone administered intravenously

  36 mg / kg peritoneal cavity, a Collison cannula was implanted in the rostral part of the third

ventricle of the cat.

  A few days after implantation of the cannula, while the cat had recovered from the operation, cerebrospinal fluid samples were collected without anesthesia, while the cat was kept free.

  in his cage, and we recorded his rectal temperature.

  The cerebrospinal fluid (csf) samples were tested immediately after collection, or after being stored at -2 for 24-48 hours, Coefficient Amount of Ulceration Group1) 6ceto-PGFa (%) (ng / ml). ..., Control -100 3.3 Substance of the invention Sample N 4 62 3.8 Sample N 7 58 3.9 Sample N 8 55 3.6

  and the PGE assay was carried out in the samples

  as in Example 9 using a radioimmunoassay kit for H-prostaglandin E

  (manufactured by the Clinical Assay Co. USA). The temperature

  rectal rature was recorded at room temperature (21 to 230C), the thermistor probe being inserted at a depth of about 10 cm into the rectum. As pyrogen, a somatic antigen of Shigella dysentriae was used, and after

  have dissolved 75 ng of pyrogen in 0.15 ml of cs.f.

  artificial pyrogen-free, the solution was injected into the third ventricle by the implanted cannula

to provoke fever.

  Two hours after administration, the rectal temperature was measured to confirm the onset of pyrexia and then forcibly injected.

  the substance of the invention (each of the samples

  Nos. 4, 7 and 8) as an aqueous solution in distilled water at a rate of 100 mg / kg, the control receiving only distilled water at this time. The rectal temperature of all animals was measured one hour after administration of the

substance of the invention.

  The c.s.f. for determination of its PGE content was collected two hours before and two hours after pyrogen injection and one hour after the administration of the substance.

of the invention.

  The results are given in Table 21.

TABLE 21

  Rectal temperature and amount of PGE in

the c.s.f.

  Temperature Quantity of Rectal Group PGE (ng / ml (C) cs.s.) Control before administration of pyrogen 39.0 ± 2.5 2 hours after adm. of 41.0 12 pyrogen 41, 1 hour after adm. of water I 40.7 10

having received the substance of the in-

  Sample N 4 before adm. pyrogen 39.0 2.6 2 hours after adm. pyrogen 41.3 1 hour after adm. sample 39.9 Sample N 7 before adm. pyrogen 38.8 2.4 2 hours after adm. pyrogen 41.1 16 1 hour after adm. of sample 39.5 Sample N 8 before adm. pyrogen 39.1 2.5 2 hours after adm. pyrogen 41.5 13 1 hour after adm. of the sample 40.0 8 I 40.0 8

Note: distilled water.

  As shown in Table 21, the rectal temperature of the cat which was increased by the pyrogen injection is reduced by administering the substance of the invention, the variation of the PGE content in the cs.s. f. presenting the same trend. Consequently, the substance is considered

  of the invention administered orally to a mammal

  has a symptom of pyrexia due to inno-

  culation of a microbial pyrogen has antipyretic activity probably related to the reduction of

of the PGE in the c.s.f.

Claims (4)

  1. Composition to regulate the production   and the metabolism of prostaglandins in mammals   characterized in that it comprises: (a) a compound of the general formula: ## STR2 ## - H-1.   wherein R denotes a member of the group consisting of radicals formed by removing OH at position 1 (alpha) or 1 (beta) arabinose, xylose, rhamnose, glucose, galactose, mannose and fructose, and R is a hydrogen atom, a C1-C4 alkyl group or a pharmaceutically acceptable metal, and (b) a pharmaceutically acceptable carrier or diluent acceptable for this one.   2. The following composition characterized in that this compound sodium p-aminobenzoate.   3. The following composition characterized in that this compound sodium p-aminobenzoate.   4. The following composition characterized in that this compound sodium p-aminobenzoate.   Claim 1 is N-D-mannoside Claim 1 is N-Dxyloside Claim 1 is N-L-rhamnoside