IPC classes for russian patent N-(4-acetoxybenzoyl)glycine lithium salt possessing tranquilising and nootropic action. RU patent 2505294. (RU 2505294):
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FIELD: medicine.
SUBSTANCE: what is described is N-(4-acetoxybenzoyl)glycine lithium salt of formula I: .
EFFECT: higher tranquilising and nootropic action.
4 tbl, 5 ex
The invention relates to new water-soluble lithium salts derived glycine (aminoacetic acid) with 4-hydroxybenzoic acid and its application as trankviliziruyuschego and nootropic drugs.
Reference that is used to recover violations of intellectual functions in patients with disorders of cerebral circulation, is pyracetam (2-oxo-1-, CAS 932-17-2). The disadvantage of pyracetam as a reference preparation is its low activity: the level of effective doses of the action is 200-500 mg/kg, the effect of pyracetam often poorly reproducible. Another drug, "Mexidol" (etilmetilgidroksipiridina succinate) shows , , anxiolytic, , , alcohol, and anticonvulsant activity etc. However, lethal dose of Mexidol, causes the death of 50% of the animals (LD 50 ) in rats is 820 (625 to 1025) mg/kg for mice 475 (365 to 617) mg/kg, while the inside is more than 3000 mg/kg in rats and 2010 (1608 to 2513) mg/kg in mice. Therapeutic index, calculated by the ratio LD 50 /ED 50 is 16,4 [Voronin, T.A. Domestic preparation of a new generation of Mexidol: the main effects mechanism of action, application. - M: 2004. - 248 S.].
Derivatives of 4- acid (parabens) used in industry as preservatives (, E-214, CAS 120-47-8, propylparaben, E-216, CAS 94-13-3), (2-()ether ethyl 3-amino-4- acid CAS 499-67-2) mestnoanesteziruyuschee [Register of medicines in Russia RLS drug encyclopedia. - 18-th vol./CH. amended .. vyshkovsky. - M: radar-MEDIA, 2009. - .681], nifuroxazide (hydrazide [(5-nitro-2-)methylene]-4-hydroxybenzoic acid) - antibacterial - blocks dehydrogenase and depresses the respiratory chain, the tricarboxylic acid cycle and a number of other biochemical processes in the microbial cell, destroys microbial wall or cytoplasmic membrane, reduces production of toxins microorganisms [Mashkovsky, PPM Medicines: manual for physicians. - T.1. - M: a New wave. - 2002. - .167]. Glycine (aminoacetic acid CAS 56-40-6) as a tool that improves mental performance and reduces emotional stress [Mashkovsky, PPM Medicines: manual for physicians. - Vol.2. - M: a New wave. - 2002. - P.124]. Lithium carbonate () used in manic phase and for the prevention of exacerbations of bipolar affective disorder, disorder, manic States of various origins, affective disorders in chronic alcoholism, drug addiction, sexual deviations, Meniere's syndrome, migraine [Mashkovsky, PPM Medicinal products: a manual for physicians. - T.1. - M: a New wave. - 2002. - .108].
The purpose of the invention consists in obtaining of highly water-soluble derivative of glycine and 4-hydroxybenzoic acid, which combines the main effects inherent in tranquilizing and neuroprotective means and is characterized by low toxicity and acting in small doses.
Summary of the invention consists in the synthesis of N-(4-) lithium formula
and use it as trankviliziruyuschego and nootropic drugs.
In studies trankviliziruyuschego and nootropic effect compounds were used doses of 10 and 50 mg/kg dissolved in izotonicescom 0.9% NaCl at room temperature. Substance is introduced once for 60 minutes before the start of the experiment. Research results subjected to statistical analysis, the significance of differences was estimated with the help of student's criterion adjusted Б.
Example 1. Lithium salt N-(4-)glycine. In the reactor, supplied with a mixer, put glycine and 50 ml DMF. Dropwise poured acid chloride 4-acetoxybenzoic acid within 1.5 hours. The reaction is carried out at cooling 1.5 hours. Then the reaction mixture was stirred for another 2.5 hours (refrigeration), controlling the pH of the medium (pH>7). To maintain protection type 6 n solution of potassium hydroxide. The resulting mixture was poured into ice and acidified with hydrochloric acid to pH=5, loose crystals of N-(4-)glycine filtered and dried. Exit 73%. T p =194-197°C, Rf 0,569 (butanol-1). M+ 237 (mass-spectrometer «Saturn-2100», Varian). Then the resulting N-(4-)glycine mixed with lithium (molar ratio 1:1) in the environment of benzene and toluene or at hashing is heated at a temperature of 100 C for 60 minutes. After cooling, the product separate filtering, washed with a small amount of solvent and dry. Get a white crystalline solid (22,2 g 97%), with a melting point above 270 degrees C, Rf 0,654 (n-propanol: water in a ratio of 7:3). M+ 243. Found: C (54,39%), H (4,09%), Li (2.85 percent), N (Of 5.79%), O (32,88%), calculated: C (54,34%), H (4,15%), Li (2.85 percent), N (USD 5.76%), O (32,90%).
Example 2. Method of the study of spontaneous motor activity to the «open field» (table 1).
This method allows to study the spontaneous locomotor activity, approximately-exploratory behavior and the level of emotional response animals. Within three minutes of observing the animals recorded the following indicators: the number of crossing squares (vertical activity), the number of on his hind legs and in the hole (approximately research activity), a number of outputs in the Central zone, the number of acts of grooming and the number of fecal bolus (emotional factor).
Example 3. A method of studying nootropic activity elaboration of the conditional reaction of passive avoidance (passive avoidance reaction).
For the study of nootropic activity compounds have used the methodology of elaboration of passive avoidance reaction. In this work, we used a modification of this technique, which was specially developed for the rats. Production of conditional reaction of avoidance of darkened compartment produced in the experimental chamber, which consisted of two adjoining compartments, large illuminated (60 x 40 cm) and a small dark (15 x 15 cm), with electrode floors. Rat was considered as trained if within 30 seconds after the training session, she had not walked in the dark compartment of the experimental chamber. Test playback memorial trail was carried out 24 hours after the study. The animal is placed in a compartment light camera and within 3 minutes to register 3 indicators: the latent period of the first entry in the dark compartment, the number of calls and total time spent in a dark cell. effect of the analyte of interest was expressed in increase of latency period of the first approach, the animal in the dark compartment, reducing the number of calls into the Bay and time of stay in it in comparison with the animals of the control group (table 3).
Example 4. The method of determining the anxiolytic effect plus - maze.
The method allows to estimate anxiolytic (, antifobicescoe) effect of psychotropic agents. Labyrinth lifted over level of the floor to a height of 70 cm and is a square pad (10 x 10 cm) with four crosswise located sleeves length of 50 cm and a width of 10 see Two sleeves are opaque fence height 40 cm, two arms are open. A rat is placed on the Central site of the maze tail to the experimenter. Within 2 minutes of observation recorded the number of visits to the open arms and the time spent in them. Increase in the number of outputs in the open arms and time of stays there, as compared to those values in the control group was assessed as a manifestation of anxiolytic () actions substances (table 2).
Example 5. Determination of acute toxicity. Acute daily toxicity after acute administration was studied in mice-female weight 25-30 Connection, in izotonicescom solution of sodium chloride animals were injected intraperitoneally in a variety of increasing doses once. Observation of animals was performed during the day, noting the number of stranded animals. Calculation LD 50 carried out by the method of Litchfield and Wilcoxon signed. Studies of acute toxicity of the proposed derivative showed that the hazard classification (harm) substances N-(4-) lithium at intraperitoneal mice belong to the class of low-toxic compounds [, I.V. Criterion of harmfulness in hygiene and toxicology when assessing the danger of chemical compounds / IV , I.P. Ulanova. - M: Medicine, 1975 - 328 S.] (table 4).
Thus, production of N-(4-) lithium dose of 10 mg/kg resulted in the manifestation expressed antiamnestic activity in the test passive avoidance reaction, which was accompanied by the increase of the latent period of the call in the dark compartment and the reduction of residence time in the dark compartment and effect ó in increasing the time of stay in the open sleeves in the test elevated plus-maze». These effects were accompanied by a reduction in acts of grooming in the «open field»test that confirms the anti-anxiety effect of N-(4-) lithium. Increasing doses up to 50 mg/kg resulted in the elimination of antiamnestic activity of this compound, while maintaining a clear trend to the manifestation of antiamnesic effect, strengthen the research and motor activity of the animals in the open field test» and the emergence of weak activity in the test «+labyrinth».
From a comparison of doses, in which the N-(4-) lithium (lithium salt N-(4-)glycine) shows and nootropic properties (10 mg/kg) with LD 50 (3751,11 mg/kg), one can conclude that the drug has a wide range of therapeutic action and low toxicity. Table 4
Comparative characteristics of effective N-(4-) lithium with Mexidol
LD 50 , mg/kg
Therapeutic index
N-(4-) lithium (intraperitoneally)
3751,11,400* (3638,84 to 3866,85) 375,1 475
Mexidol (intraperitoneally)
(365 to 617) 16,4 Attitude 14,42
* - the data are statistically valid, student's criterion adjusted Б, p<0.05
Water soluble lithium salt N-(4-)glycine formula
with tranquilizing and nootropic effects.
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