JPH0222047B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a prostaglandin regulator, particularly an antithrombotic agent, which contains an aminobenzoic acid derivative represented by the following general formula () as an active ingredient. Prostaglandins (hereinafter referred to as PGs) have various physiological effects in the body and have recently been recognized as important. However, some types have short half-lives and are difficult to extract in pure form, while others are unstable and have many problems when used as medicines. On the other hand, there is a method of regulating prostaglandins produced in the body using regulating agents. An example of this is aspirin (Nature 239, 33-34, 1972).
However, aspirin is an inhibitor of cyclooxygenase, which is involved in the early stages of PGs metabolism, and is therefore positioned as a blocker for all PGs. In other words, it can be said to be a production inhibitor for all PGs, but it does not have the effect of increasing the production of certain PGs, and has its own limitations as a PGs regulator. Furthermore, aspirin causes gastrointestinal disorders as a side effect, making it difficult to administer it for a long period of time. Therefore, the present inventors conducted extensive research into compounds with few side effects and strong prostaglandin regulating effects, particularly strong antithrombotic effects, and found that aminobenzoic acid derivatives represented by the general formula () are effective. I've reached it. (However, in the formula, R ₁ represents a monosaccharide residue, and R ₂ represents a hydrogen atom, a pharmaceutically acceptable metal, or a C ₁ to _{C 4} lower alkyl group). Although the compound represented by the above general formula () in the present invention (hereinafter referred to as the "substance") has a simple structure, it has extremely low toxicity and has no antibacterial activity, so there is no concern about disturbance of intestinal flora. It can be used continuously for a long period of time. Furthermore, this substance does not have mutagenicity or affect cellular or humoral immunity, and therefore does not pose any risk of teratogenicity or allergic reactions to healthy people, making it an extremely safe drug. In the above formula (), R ₁ represents a monosaccharide, and these sugars can be in the form of D-form, L-form, α-form, β-form, or a mixture thereof. Therefore, the substance can be in the alpha or beta form or a mixture thereof. Examples of the sugars mentioned here include the following. arabinose, xylose,
These include mannose, glucose, galactose, fructose and rhamnose. In the general formula (), R ₁ is particularly preferably a monosaccharide residue with one hydroxyl group removed from the reducing end. In the general formula (), R ₂ is a hydrogen atom, a pharmaceutically acceptable metal such as alkali metals such as Na and K, alkaline earth metals such as Ca and Mg, and aluminum metal, or methyl, ethyl, propyl, butyl. Indicates a lower alkyl group such as. The method for producing this substance and the physicochemical properties of the substance are disclosed in JP-A-54-132239, JP-A-55-92320, and JP-A-56-55397. Next, the toxicological properties of this substance are shown. (1) Acute toxicity Acute toxicity was investigated by forced oral administration using ICR-JCL mice. This substance was dissolved or suspended in distilled water, and the solution was adjusted to a predetermined amount using a stomach tube and administered. After administration, the symptoms of toxicity were continued to be observed, and the LD ₅₀ value was determined from the mortality rate over time up to the 7th day. The findings were obtained through autopsy in both surviving and dead cases. _LD50 value is Litchfield-Wilcoxon (Litchfield-Wilcoxon)
(Wilcoxon) graphic calculation method. Table 1 shows the results.
Shown below. All of them have large LD ₅₀ values, and it is clear that all of them are substances with extremely low toxicity.

[Table] (2) Effects on the gastrointestinal tract.Administer 5 g of this substance or aspirin as a suspension of PH3 to beagle dogs weighing 9 to 12 kg.
Gastric bleeding was examined 90 minutes after administration. Bleeding was observed with aspirin, but not with this substance. This substance is a low toxicity substance and is an extremely safe drug. Next, we will show the pharmacological properties of this substance. The following shows that this substance regulates the production of PGs and its metabolites. (1) Platelet aggregation is mainly caused by PGG ₂ , PGH ₂ ,
It is known to be induced by TXA ₂ ,
This substance was confirmed to have a platelet aggregation inhibitory effect. (See Examples 1 and 3) (2) Regarding malondialdehyde (MDA), which is known as one of the PGs metabolites, it was confirmed that this substance has the effect of suppressing the production of platelet MDA. (See Example 2) (3) PGI ₂ using in vitro cultured cells
It was confirmed that this substance has a promoting effect on production. (See Example 3) Generally, PGs are found in various organs throughout the body.
It has an effect closely related to the function of that organ.
Furthermore, the physiological and pharmacological effects of PGs have already been found to be extremely wide-ranging. That is, as a pharmacological action on the circulatory system, it mainly acts on dilation and constriction of blood vessels, PGE and PGI have a large dilation effect, and TXA conversely constricts arteries.
These effects result in increases and decreases in blood pressure, and furthermore, they have therapeutic effects on angina pectoris and arrhythmia. Furthermore, if platelet aggregation progresses excessively, it will cause arteriosclerosis, cerebral infarction, myocardial infarction, and stroke, and PGs have a large effect on platelets. That is,
PGD, PGI, and PGE have an antagonistic effect on this platelet aggregation effect, while TXA ₂ , PGG ₂ , PGH ₂ , and PGE ₂ have an inducing or promoting effect. Therefore, it is clear that the above-mentioned diseases can be treated and prevented by making these adjustments. This substance has low acute toxicity and few other side effects, so it is useful as a medicine for animals and even humans.
As a medicine, it is used for humans as an antithrombotic agent. Next, we will discuss the formulation of this substance. When this substance is used as an antithrombotic agent, it can be used in any form convenient for obtaining the medicinal effect depending on the type and symptoms of the disease, and may be used alone or in a mixture with a pharmaceutically acceptable diluent and other drugs. Can be used. The substances can be provided in dosage unit form.
Contains an effective amount of the active ingredient, and its forms include powders, granules, tablets, sugar-coated tablets, etc. for oral use.
Capsules, syrups, pills, suspensions, liquids,
Emulsions, etc. For parenteral use, it can be in the form of an ampoule or bottle as an injection solution. Suppositories are also available. The diluent may be solid, liquid, or semi-solid;
Examples include: That is, excipients, fillers, binders, wetting agents, disintegrants, surfactants, lubricants, dispersants, buffers, fragrances, preservatives,
These include solubilizing agents and solvents. Specific examples include lactose, sucrose, sorbitol, mannitrate, starch, precipitated calcium carbonate, heavy magnesium oxide, talc, calcium stearate, magnesium stearate, cellulose or its derivatives, amylopectin, polyvinyl alcohol, gelatin, surfactant, water,
These include physiological saline, ethanol, glycerin, propylene glycol, cacao butter, lauric fat, petrolatum, paraffin, and higher alcohols. The bioactive agent of the present invention can be produced by any known method. The active ingredient in the compositions used in the present invention generally comprises from 0.01% to 100wt.%, preferably from 0.05% to 80wt.%. The bioactive agent of the present invention can be administered orally or parenterally to humans and animals, but oral administration is preferred.
Oral administration includes sublingual administration. Parenteral administration includes injection administration (eg, subcutaneous, intramuscular, intravenous injection, infusion), rectal administration, and the like. The dosage of the bioactive agent of the present invention depends on whether it is an animal or a human, and is influenced by age, individual differences, medical conditions, etc. Therefore, in some cases, doses outside the range shown below may be administered, but in general, humans are administered. In this case, the oral dosage of this substance is 0.1 to 500 mg, preferably 1 to 250 mg per day per 1 kg of body weight, and the parenteral dosage is the same.
0.01-200mg, preferably 0.1-100mg once to 4 times
Administer in divided doses. Hereinafter, the present invention will be explained in more detail by showing formulation examples and manufacturing examples of the substance of the present invention. Parts in the examples below indicate weight. Example 1 Effect of inhibiting platelet aggregation due to the PGs regulating effect of this substance It has been found that platelet aggregation is mainly induced by arachidonic acid metabolites PGG ₂ , PGH ₂ , and TXA ₂ . Therefore, the effect of this substance on platelet aggregation was investigated as follows. A titrate solution for blood sedimentation was used as an anticoagulant, and 9 samples of blood were collected from each sample. This is 400
×g, centrifugation for 6 minutes, collect the supernatant, and
PRP (Platelet Rich Plasma) was prepared. The residue after collecting the supernatant was further centrifuged at 700×g for 20 minutes, and the supernatant was used as PPP (Platelet Poor Plasma). This change in the transmittance of PRP was measured using an aggregometer (PAP-3 type, manufactured by Biodata), and the degree of platelet aggregation was determined. Arachidonic acid (1.64mM), adenosine diphosphate (50μM) as flocculants,
This substance (sample no.
1 to 8) were each added to the PRP 2 minutes before adding the flocculant. The results are shown in FIGS. 1 to 6, and the present substance showed an inhibitory effect on platelet aggregation against all aggregating agents. This means that this substance contains PGs, especially PGs, especially PGG ₂ ,
This shows that it regulates the production of PGH ₂ and TXA ₂ . Example 2 Effect of suppressing platelet MDA production due to the PGs regulating effect of this substance PRP (Platelet Rich Plasma) is collected from human blood, and this PRP is further centrifuged and washed to prepare washed platelets. After adding a predetermined amount of this substance (sample No. 4, 7, 8) to the washed platelets, Ca-ionophore A-23187 was added and incubated at 37°C for 5 minutes, and the coloring agent thiobarbituric acid ( Thiobarbituric Acid) was added, and this was extracted and separated using a methanol-butanol mixed solution, and the resulting mixture was colorimetrically determined at 535 nm. The results are shown in Figure 7 _{, and this substance has an effect on the production of MDA using mouse 3T3 fibroblasts capable of producing PGI 2 .} The effects of this substance were investigated. Arachidonic acid, a precursor of PGs, was added to the 3T3 fibroblast culture medium and incubated at 37°C for 5 minutes to produce PGI _2.
was produced and used as a control. On the other hand, 30mM of this substance (sample Nos. 1, 3,
4,7-11) was added to 4 ml of 3T3 culture solution and incubated for 2 minutes, then arachidonic acid was added and cultured in the same manner as the control.
Those that produced PGI ₂ were designated as the group treated with this substance.
Using the culture supernatants of both groups, their PGI ₂ production ability was compared using the platelet aggregation inhibitory effect as an index.
Arachidonic acid (2.43mM) as a platelet aggregating agent
was used. The results are shown in Figure 8. In the culture medium for 5 minutes after addition of arachidonic acid as a precursor of PGs, although inhibition of platelet aggregation was observed in the control, a marked inhibition was observed in the group of this substance, and this substance inhibited PGI. ₂
It was found that the production of Example 4 Inhibitory effect on thromboembolism death due to PGs regulating effect of this substance 300 mg/Kg of this substance was orally administered to mice, and 3 hours later ADP or collagen was intravenously injected to measure the mortality rate. As a control, distilled water was orally administered instead of this substance. The results are shown in Table-2. All of the compounds tested suppressed mortality, but especially p-
It is noteworthy that sodium aminobenzoate-ND-mannoside exhibits excellent effects.

【table】

[Table] Formulation example 1 This substance (p-aminobenzoic acid-N-D-galactoside-Na salt) 10 (parts) Heavy magnesium oxide 15% Lactose 75% is uniformly mixed and powdered or finely divided into 350μ The following powder is used. This powder was also put into a capsule container to make a capsule. Formulation example 2 This substance (p-aminobenzoic acid-N-D-xyloside Na salt) 45 (parts) Starch 15 Lactose 16 Crystalline cellulose 21 Polyvinyl alcohol 3 Water After uniformly mixing, crush and granulate. It is dried and then sieved to form granules with a size of 1410μ to 177μ. Formulation Example 3 p-Aminobenzoic acid-N- in Formulation Example 2
Granules were made in the same manner using o-aminobenzoic acid ester-N-L-rhamnoside instead of D-mannoside Na salt, and 4 parts of calcium stearate was added to 96 parts of the granules, and the mixture was compression molded to give a diameter
Take a 10mm tablet. Formulation Example 4 10 parts of crystalline cellulose and 3 parts of calcium stearate were added to 90 parts of the granules obtained by the method of Formulation Example 2, and compression molded to form tablets with a diameter of 3 mm, which were mixed with syrup gelatin and precipitated calcium carbonate. Add the suspension to make sugar-coated tablets. Formulation example 5 This substance (m-aminobenzoic acid-N-L-rhamnoside Na salt) 0.6 (parts) Nonionic surfactant 2.4 Physiological saline 97 After heating and mixing, place in an ampoule, sterilize, and prepare as an injection. do.

[Brief explanation of drawings]

Figure 1 shows the effect of this substance on arachidonic acid aggregation, Figure 2 shows the effect of this substance on adenosine diphosphate aggregation, and Figure 3 shows the effect of this substance on collagen aggregation. Figure 4 shows the effect of this substance on epinephrine aggregation, and Figure 5 shows the effect of the substance on epinephrine aggregation.
Showing the effect of this substance on ristocetin aggregation,
Figure 6 shows the effect of this substance on thrombin aggregation, Figure 7 shows the effect of this substance on platelet MDA production, and Figure ₈ shows the effect of this substance on thrombin aggregation.
It is a figure showing the influence of this substance on production.

Claims (1)

[Claims] 1 General formula () (wherein R ₁ represents a monosaccharide residue and R ₂ represents a hydrogen atom, a pharmaceutically acceptable metal, or a C ₁ to C ₄ lower alkyl group) Antithrombotic agents.