JPS60172929A - Agent for alleviating amnesia - Google Patents

Abstract

PURPOSE:An agent for alleviating amnesia containing N-benxyl-oxycarbonyl-glycyl-L-prolyl-methane derivative as an active ingredient. CONSTITUTION:A safe agent for alleviating amnesia having low toxicity, containing N-benzyloxycarbonyl-glycyl-L-prolyl-chloromethane shown by the formula I or N-benzyloxycarboxyl-glycyl-L-prolyl-diazomethane shown by the formula II as an active ingredient. Administration can be carried out by various injections such as intravenous injection, hypodermic injection, intramuscular injection, etc. or orally. A dose is 1-900mg/day, especially preferably 5-500mg/ day in case of oral administration, 0.5-500mg/day, especially preferably 1- 200mg/day in case of intravenous injection. >=0.1wt%, preferably 1-50wt% active ingredient compound is contained in the agent for alleviating amnesia.

Description

[Detailed description of the invention] The present invention relates to a novel anti-amnestic agent.

More specifically, the present invention provides an anti-inflammatory drug characterized in that N-benzyloxycarbonyl-glycyl-L-prolyl-chloromethane contains N-benzyloxycarbonyl-glycyl-L-prolyl-diazomethane as an active ingredient. Concerning amnesia drugs.

Although much remains unclear about the mechanisms of memory, it has recently been revealed that ptedohormones such as pasopressin and TRH (thyrotropin releasing hormone) are involved in memory. In other words, when these putidohormones are administered to rats with experimental amnesia, clear memory recovery is observed, and it has been confirmed that they have similar effects in clinical trials on literary figures (N
euroscience, vol. 4, 1529-1537
Baichiji, 1979: 5science, vol. 211,
601-603 Baichiji, 1981). All of these peptide hormones contain proline and have been shown to exist in the brain. It has long been assumed that these proline-containing ptade hormones are metabolized by endogenous butidase, but their existence was actually confirmed in the early 1970s. post-sexual endopeptidase (post-
Due to its specificity, it is presumed that much of the zoroline-containing enzyme (proline cleaving enzyme) is involved in the metabolism of putidohormone.

Searches for inhibitors against proline-specific endopeptidases have been conducted in a wide variety of ways, including endogenous ones. Among these searches, N-benzyloxy carbonyl-k IJ sil- L-prolyl-chloro Methane and Eq.

N-benzyloxycarbonyl-f! J
Sik-L-prolyl-diazomethane has been shown to be a potent inhibitor (Biochem
istry, Vol. 6, No. 13, 2492-2498, 1977].

As a result of various studies, the present inventors have found that N-benzyloxycarbonyl-glycyl-L-prolyl-chloromethane and N-benzyloxycarbonyl-glycyl-L-prolyl-diazomethane each exhibit excellent anti-amnestic effects. I found out. It was discovered for the first time by the present inventors that these compounds exhibit excellent anti-amnestic effects, and the present invention is based on this finding.

As a result of a detailed study of the effects of each of these compounds on passive avoidance learning in rats, the present inventors found that these compounds exhibit remarkable anti-amnestic effects when administered intraperitoneally and orally, and have low toxicity. It was found that it is a safe substance.

Accordingly, the present invention provides an anti-amnestic agent containing N-benzyloxycarbonyl-crysyl-L-prolyl-chloromethane or N-benzyloxycarbonyl-glycyl-L-prolyl-diazomethane as an active ingredient. This is what we provide.

The present invention will be explained in detail below.

First, the anti-amnestic effects and toxicity of these compounds confirmed by the present inventors will be described.

1. Anti-amnestic effect The method for testing anti-amnestic effect is Int, 8ymp, o
nPharmacology of Learning
Kubota and Memory (1982),
The effect of the compound on passive avoidance behavior was measured according to the method of Hayashi et al. The passive avoidance test box used in this test had a grid-like electrode bed (15 cm long, 30 cm wide).
) and the stand placed behind it on the right (vertical 156 rIL, horizontal 15
11! I+! , 4 m high) and 1.7 m high on the floor.
It is a box made of transparent plastic that allows the flow of electric current A and allows the inside to be observed.

Experimental Example 1 Effect of N-benzyloxycarbonyl-glycyl-L-prolyl-chloromethane Male Wistar SLC rats (body weight +00~15
0.9) was placed on the platform of a passive avoidance test box, and when the rat landed on the floor, a current was applied and continued until the rat climbed onto the platform.

Once the rat remained on the platform for more than 15 seconds, it was considered to have learned and was removed from the box. The number of electric shocks is usually 2~
It was 6 times. or.

To avoid unevenness of the rats, first f! 1 to get bored
Those that took more than 0 seconds or those that required descending to the floor more than 5 times were excluded. For the rats that underwent learning, 0.5 t CMC solution was administered intraperitoneally to the control group of the first group, and 5 mt/h of scopolamine hydrobromide was administered intraperitoneally to the other groups. It was created. N-benzyloxycarbonyl-crisyl-L-,-1/lolyl-chloromethane and aniracetam (1-p-7 nisoy-2-pyrrolidinone) as a control were administered intraperitoneally at the doses shown in Table 1 one hour after the end of the study. Or administered orally. All of the drugs used were dissolved or suspended in a 5% CMC solution. The results of these tests are shown in Tables 1 and 2, and it showed an anti-amnestic effect equivalent to that of aniracetam when administered intraperitoneally, and a stronger anti-amnestic effect than the control when administered orally.

Experimental Example 2 Effect of N-benzyloxycarbonyl-glycyl-L-prolyl-diazomethane The first group of Con7
Experimental amnesic rats were created by intraperitoneally administering 0.15 q6 CMC bath solution to the Tofour group, and 3 ml of scopolamine hydrobromide to the other five groups. N-benzyloxycarbonyl-glycyl-L-prolyl-diazomethane was administered intraperitoneally or orally at the doses shown in Table 3 one hour after the end of the study. All drugs used were 0.
It is dissolved or suspended in 5-CMC solution. The test results are shown in Table 6, and showed a stronger anti-amnestic effect than the control group when administered intraperitoneally and orally.

2. Toxicity Test Experimental Example 3 LD 50 was determined using five 4-week-old ICR male mice weighing 15 to 20 I. Mice were treated with N-(benzyloxycarbonyl-glycyl-L-prolyl-chloromethane and N-benzyloxycarbonyl-glycyl-L), respectively.
-Prolyl-diazomethane was administered orally as well as intraperitoneally, and the LD 50 was calculated. Next, the results of these tests are shown in Table 4.

The above test results show that each of the above compounds has an excellent anti-amnestic effect and can be used as a low toxicity drug for the treatment of memory disorders such as senile dementia.

Administration of N-benzyloxycarbonyl-glycyl-L-prolyl-chloromethane and N-benzyloxycarbonyl-glycyl-L-prolyl-diazomethane includes various injections such as intravenous injection, subcutaneous injection, and intramuscular injection, or oral administration. Although it can be carried out by various methods, oral administration and intravenous administration are particularly preferred. The dosage of each of these compounds is preferably in the range of 1 to 900 μ per day, especially 5 to 500 μ per day in the case of oral administration, and 0.5 to 500 μ per day, especially 1 to 500 μ per day in the case of intravenous administration. 20
0■ is preferable.

The anti-amnestic agent of the present invention can be prepared by arbitrarily selecting methods commonly used for preparing various preparations according to various dosage forms. For example, if an oily substance is
Suitable examples include forms suitable for absorption from the intestinal tract, such as soft capsules and oral liquid preparations. Soft capsules for oral administration are a fixed dosage form, containing a solubilizing agent such as macrogol, phlopylene glycol, a preservative such as ether paraoxybenzoate, propyl paraoxybenzoate or sorbic acid, and optionally It may contain flavoring agents, dyes, fragrances, etc.

Examples of oral liquid preparations include aqueous or oily suspensions, solutions, syrups, and elixirs. Such liquid preparations may contain commonly used additives, such as suspending agents such as sorbitol syrup, methyl cellulose, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel or hydrogenated edible fats, emulsifying agents such as lecithin, monooleate, and rubitan. Or gum arabic.

Non-aqueous vehicles such as almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol, preservatives such as methyl roseoxybenzoate or propyl roseoxybenzoate, sorbic acid, optionally containing dyes, fragrances, etc. good.

Injectable preparations can be packaged in fixed-dose ampoules or in multi-dose, four-dose containers with additives such as preservatives and solubilizing agents.

The formulations may take any form such as suspensions, solutions, emulsions in oily or aqueous vehicles.

It may also contain additives such as suspending agents.

The anti-amnestic agent of the present invention usually contains the above-mentioned active ingredient compound in an amount of 0.1 or more, preferably 1 to 50, depending on the various dosage forms.

Next, specific formulation examples are listed, but the formulation of the anti-amnestic agent of the present invention is not limited to the following examples.

Formulation Example 1 Injection N-benzyloxycarbonyl-glycyl-10-L-
Prolyl-chloromethanesorbitan monostearate 2g Propylene glycol 60~ Purified soybean lecithin 2■ Cholesterol 1■ Glucose 50■ Distilled water 1ml (manufactured) N-benzyloxycarbonyl-glycyl = L-prolyl-chloromethane, hydrogenated castor oil polyoxyethylene 6
Mix and melt 0 mole ether, sorbitan monostearate, phlopylene gellicol, purified soybean lecithin, and cholesterol on a water bath at approximately 80°C to make a homogeneous mixture. Add distilled water at 80°C while stirring and discontinue stirring. After solubilizing, add glucose and make 1- with distilled water. After sterile filtration, the mixture was filled into brown ampoules and thawed.

Formulation Example 2 Soft Capsules Macrogol 400 350η Propylene Glycol 68 Dipotassium Quiglycyrrhizinate 1■ Peppermint Oil 1■ Gelatin 122 Quiglycerin 6o, 5■ D-Sorbitol Solution 12.2■ Ethyl Roseoxybenzoate 0.8~ Pa2oxy Propyl benzoate 0.5 kg (manufacturing) N-benzyloxycarbonyl-glycyl-L-prolyl-diazomethane, Macrogol 400, glycyrrhizinate dicaryuno, peppermint oil, and propylene glycol are mixed evenly to prepare a suspension. Next, gelatin, glycerin, D-sorbitol solution 1. e. Prepare a soft capsule coating agent using ethyl paraoxybenzoate and propyl paraoxybenzoate. These are used to form and fill soft capsules.

N-penlyloxycarbonyl-glycyl-L-prolyl-diazomethane and N-benzyloxycarbonyl-glycyl-L-prolyl-chloromethane used in the anti-amnestic agent of the present invention are both known substances, Although they are easily available, examples of preparations for each compound are illustratively shown below.

/) Preparation example of N-benzyloxycarbonyl-crysyl-L-zorolyluriazomethane N-benzyloxycarbonyl-glycyl-L-zoroline (Serpa Fine
Biochemicals (5erva F'ine Hioc
Chemicals), US, 6>, 2vy is dissolved in tetrahydrone ran/3Qml, and after cooling to -/0°C, triethylamine//ml and ethyl chloroformate grrLt are added. Next, 2t of a cooled ethereal solution of diazomethane is added and kept at 0°C for 10 minutes. The reaction solution is washed with 200 ml of O, OiN vinegar and saturated sodium hydrocarbon solution, and the ether layer is separated. At this time, V was dried with crushed magnesium and allowed to stand, and the ether was distilled off to obtain an oily substance.

Sound collection 1g, 3tc yield gs%) J) N-benzyloxycarbonyl-glycyl-L-
To illustrate the production of prolyl-chloromethane, add 3M hydrochloric acid in ethanol, keep at 0°C with a trowel, and immediately wash with 200mL of /N salt solution and 200mL of saturated sodium bicarbonate. The ether layer was separated, dried over valent magnesium, filtered, and then the ether was distilled off to obtain an oily substance.

Collection i1 /9. ! ;f (yield?, t%) Patent applicant: Yakult Honsha Co., Ltd.

Claims (1)

[Claims] N-benzyloxycarbonyl-glycyl-L-prolyl-chloromethane or N-inzyloxycarbonyl-
An anti-amnestic agent characterized by containing glycyl-L-prolyl-diazomethane as an active ingredient.