RU2476424C1 - N-(3-chloro-4-propyloxyphenyl)-3,5-dichlorosalicylamide having anthelmintic activity - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: disclosed is a novel biologically active salicylanilide derivative and specifically

N-(3-chloro-4-propyloxyphenyl)-3,5-dichlorosalicylamide.

EFFECT: obtaining a compound having high anthelmintic activity with low toxicity, which enables to use the disclosed compound in medicine and veterinary.

1 cl, 3 ex, 1 tbl

Description

The invention relates to organic chemistry and relates to a new biologically active substance, and specifically a salicylanilide derivative of the formula 1 (C-1):

,

,

possessing anthelmintic activity.

Salicylanilides with antifungal and bactericidal activity are known [see, for example: Pat. 2,703,332 US. Poly Halo-salicylanilides; Pat. 2.802.029 US. Bromsalicyloyl-chloranilide; Pat. 4.310.682 US. Halo-salicylanilide].

Salicilanilides have been claimed for the treatment of diseases in which an increase in mitochondrial respiration is useful [RF Application 2006116421, 8 IPC C07C 235/64. Salicylic acid anilides; withdrawn].

A class of aromatic derivatives of salicylanilides with various substituents, their salts, including amines, effective against the parasite of the hepatic hepatic, is described [Pat. 3.914.418 US. Method of controlling liver fluke infection].

Compositions with salicylanilides for controlling trematodes and nematodes are proposed [Pat. 3.989.826 US. Method of killing internal Parasites using salicylanilides].

Known antibacterial agent, which is a 3,5-dichlorosalicylic acid, previously used as an intermediate in the production of antibacterial agents [Pat. 10,81173 DE. Antibacterial detergent compositions; GB 848,306 (A)].

Known substituted salicylanilides having anthelmintic activity [Pat. 4.025.647 US. Salicylanilide derivatives having anthelmintic activity].

Markush structures for salicylanilides given in the above documents and other sources available to the authors do not include the compound proposed by the authors.

At present, the anthelmintic drug NIKLOSAMID (Germany) is widely used in medicine and veterinary medicine [see, for example: Pat. 3,074,848 US; claims priority, application Germany 1960.02.02. 2-Hydroxy-5,2'-dichloro-4'-nitrobenzanilide for combating tapeworms; Pat. 2557615 DE, Int. Cl. ² A61K 31/165, 47/00. Neue Niclosamide Suspensionsfor-mulierungen]. In Russia, the drug of the same active substance is known under the trade name FENASAL (USSR) [Lebedeva M.N., Mikhaylitsyn F.S., Sergiev V.P. The creation of domestic antiparasitic drugs [Text] / MN Lebedev and [other] // Med. parasitology and parasitic diseases. 2001. No4. S.13-16; RF patent 1166370, 7 IPC A61K 9/16. Anthelmintic agent; RF patent 2276983, 8 IPC A61K 31/609, A61K 9/08, A61K 47/46, A61P 33/10. Cestodocidal composition for animals].

Phenasal (Phenasalum) - 5,2'-Dichloro-4'-nitrosalicylanilide or N- (2-chloro-4-nitrophenyl) -3-chlorosalicylamide of the formula 2 (C-2):

Synonyms: Yomezan, Yomesan, Aten, Atenase, Biomesan, Cestocid, Copharten, Devermin, Grandal, Helmiantin, Jometan, Kontal, Lintex, Niclosamidum, Radeverm, Teniarene, Vermitin, Yomesan, etc. completely denaturing the structure of cells, paralyzes the neuromuscular system, destroys its integumentary tissues, disrupts the energy exchange of helminths. FENASAL is a non-toxic substance (class IV), its preparations do not exhibit embryotropic, carcinogenic properties.

The disadvantages of the drug include the laborious synthesis of the intermediate product FENASAL 5-chlorosalicylic acid, for which chlorine gas is used, and the completeness of the reaction is controlled by the weight gain of the reaction mixture.

Previously, the authors synthesized N- (3-chloro-4-methylphenyl) -3,5-dibromosalicylamide of the formula 3 similar in principle to FENASALU:

Compound 3 (C-3) under the conditional name RONTANOKS was tested on farm animals - sheep, cattle (cattle) in the All-Russian Research Institute of Helminthology named after K.I.Skryabin (VIGIS, Moscow) [Sevbo D.P. Anthelmintic agent based on N- (3-chloro-4-methylphenyl) -3,5-di-bromosalicylamide: RF patent No. 2370484 / D.P. Sevbo, E.I. Sakanyan, S.N. Trusov and others; Trusov, S.N. The spectrum of anthelmintic activity of RONTANOKSA / S.N. Trusov, F.S.Mikhailitsyn, D.P. Sevbo, I.A. Arkhipov, A.V. Rodionov, N.I. Koshevarov // Medical parasitology and parasitic diseases. 2007. No4. S.50-51].

The disadvantages of C-3 include the fact that it is a more toxic substance (LD ₅₀ = 178 mg / kg) with intraperitoneal administration compared with FENASAL [Buryakina A.V. Acute toxicity of rontanox with anthelmintic activity / A.V. Buryakina, F.S. Mikhaylitsyn, D.P. Sevbo, A.Yu. Malakhova, S.N. Trusov // Russian Parasitological Journal. 2011. No1. S.108-111]. It should be noted that anthelmintic activity is evaluated by oral administration (Example 3), however, with a similar administration, acute toxicity (LD ₅₀ ) for various halogen-substituted phenylsalicylamides, including phenasal, cannot be calculated due to the limitation of the introduction of maximum allowable volumes of liquid. Therefore, for a comparative assessment of acute toxicity (LD ₅₀ ) of halogen-substituted phenyl salicylamides, the route of administration was changed to intraperitoneal in 1% starch paste (example 2).

In order to reduce toxicity, alkoxyanalogues of rontanox were synthesized - compounds 4 and 5 (C-4; C-5):

R = -CH ₃ (compound 4); —CH ₂ —CH ₂ —CH ₃ (compound 5).

Both compounds exhibit a higher anthelmintic activity in case of hymenolepidosis in comparison with phenasal [Sevbo D.P. Obtaining and studying the antihymenolepidoid activity of the compound MST-13 / D.P. Sevbo, S.N. Trusov, A.Yu. Malakhova, G.A. Gitsu, F.S. Mikhaylitsyn // Medical parasitology and parasitic diseases. 2010. No2. S.45; Sevbo D.P. Obtaining and studying the antihymenolepidoid activity of the compound MST-25 / S.N. Trusov, F.S. Mikhaylitsyn, D.P. Sevbo, A.Yu. Malakhova, V.Yu. Podushkin // Medical parasitology and parasitic diseases. 2011. No2. P.43], but have lower toxicity compared to RONTANOX (C-4: LD ₅₀ > 250 mg / kg; C-5: LD ₅₀ > 700 mg / kg).

Compound 5 (prototype) is the closest in structure and activity to the claimed compound 1 (C-1).

The objective of the invention is to increase the anthelmintic activity in case of hymenolepidosis while maintaining a sufficiently low toxicity and expanding the arsenal of means of influence on a living organism.

The task is implemented by a new derivative of salicylanilide of the formula 1.

Compound 1 is prepared as follows.

Example 1. Obtaining N- (3-chloro-4-propyloxyphenyl) -3,5-dichloro-salicylamide.

1.1. Obtaining 3-chloro-4-propyloxyaniline:

To a solution of 10 g of 3,4-dichloronitrobenzene in 50 ml of propanol-1 at a temperature of from 40 to 45 ° C and stirring for five hours, a solution of sodium propylate obtained from 1.2 g of sodium metal and 20 ml of propanol-1 is gradually added. The completeness of the reaction is controlled by determining the pH of a sample of the reaction mixture with water 1: 1 (pH should be in the range of 7.5 to 8). The criterion for the end of the reaction is the absence of 3,4-dichloronitrobenzene during chromatographic control of the reaction mass. After the reaction, the mass is filtered off from the residual sodium chloride and propyl alcohol is distilled off under vacuum. To the remaining thick yellow-orange liquid was added 20 ml of toluene and again distilled off under vacuum. The product is an oily mass. According to GLC analysis with a mass selective detector, the mass spectrum, m / z 215 (M +), corresponds to 3-chloro-4-propyloxy nitrobenzene with a content of at least 96.5%. The contents of the container are dissolved in 50 ml of toluene, this solution is mixed with 50.0 g of cast iron chips and a solution of 35 g of ammonium chloride in 100 ml of water and heated with stirring for 6 hours. The end of the reaction is determined by TLC by the disappearance of the original nitro derivative in the toluene layer. Then the mass is cooled, the toluene layer is separated, and the remaining reaction mass is filtered under vacuum, the toluene solution of the amino derivative is separated from the aqueous layer on the funnel. The resulting toluene extracts are combined and filtered into a flask for distillation. The toluene is removed in vacuo, and the residue is transferred to boiling benzene in a glass for crystallization. The precipitate formed is filtered off, washed with benzene and dried at 40-50 ° C, and 5.14 g of 3-chloro-4-propyloxyaniline are obtained, which is chromatographically homogeneous with mp. 52-53 ° C. The total yield in stages is 65.0%.

The analysis is performed by GLC with a mass selective detector. Mass spectrum, m / z: 185 (M ⁺ ), corresponds to the obtained compound.

1. 2. Obtaining N- (3-chloro-4-propyloxyphenyl) -3,5-dichlorosalicylic amide (C-1):

To a mixture of 6.8 g of 3,5-dichlorosalicylic acid, 150 ml of toluene, 3.4 g of 3-chloro-4-propyloxyaniline, 0.96 ml of phosphorus trichloride was added. The reaction mass is boiled for 2 hours, then evaporated to 1/2 volume, and the residue is poured into a glass for crystallization. The precipitate formed is filtered off and washed with a 5% sodium hydrogen carbonate solution to remove residues of 3,5-dichlorosalicylic acid. Obtain 4.5 g of substance C-1 (yield 66%).

Thus, from the known 3,5-dichlorosalicylic acid and 3-chloro-4-propyloxyaniline, phenyl salicylamide (C-1) is obtained with mp. 153-154 ° C (from benzene), which contains at least 96% of the basic substance (according to HPLC).

HPLC analysis was performed on a Shimadzu LC-20 instrument in gradient elution with a 0.1% phosphoric acid-acetonitrile mixture on a Zorbax Eclips SB-C182.1 * column with parameters: 155 mm, 5 mkm, 40 ° C, autosampler, SU- 20A, sample volume 10 μl. The retention time under these conditions was 16.4 minutes, characteristic of most phenylalkylamides. In the IR spectrum (shot in potassium bromide tablets) of this sample, there are absorption bands characteristic of phenolic hydroxyl ν _о-н = 3420 cm ^-1 ; ν _NHamide = 3310 cm ^-1 ; ν _{C = Oamide} = 1630 cm ^-1 . The PMR spectrum recorded on a ¹ N NMR instrument (400 MHz, DMSO) completely corresponds to the structure (C-1): δ = 12.92 ppm (s, 1H) from phenolic hydroxyl; δ = 10.55 ppm (s, 1H) NH amide; δ = 8.12 ppm (d, J = 2.0 Hz 1H) and δ = 7.8 ppm. (d, J = 2.2 Hz 1H) - two metaprotons of 3,5-dichlorosalicylate; δ = 7.7 ppm (s, 1H), δ = 7.57 ppm. (dd, J = 8.9; 2.2 1H), δ = 7.12 ppm. (d, J = 9.0 Hz, 1H) - three protons of phenylamide; δ = 4.02 ppm (t, J = 6.3 Hz, 2H), δ = 1.86-1.75 ppm. (m, 2H), δ = 1.05 ppm. (t, J = 7.4 Hz, 3H) are the protons of the propyloxy group.

The salicylamide (C-1) thus obtained, which is a white powder, soluble in dioxane, moderately in alcohol, toluene, insoluble in water, was directly used to determine acute toxicity and anthelmintic activity (see examples 2, 3).

Pharmacological studies.

Example 2. Assessment of acute toxicity.

The determination of acute toxicity (calculation of a single semi-lethal dose - LD ₅₀ ) was carried out in mice by the express method of Prozorovsky [Prozorovsky VB The express-method for determining the average effective dose and its error / VB Prozorovsky, MP Prozorovskaya, VM Demchenko // Pharmacology and Toxicology, 1978. No. 4. C.497-502].

The term "effective dose" refers to the amount of an agent for treating or preventing a condition that can be treated by administering a composition based on a given substance. The indicated amount is an amount sufficient to exhibit a noticeable therapeutic, prophylactic or alleviating effect. The specific effective amount for the subject will depend on the body weight, nature and severity of the condition to be treated. For correct comparison with the base drug, the proposed tool was asked once in comparable doses from 0.5 g / kg to 0.063 g / kg.

Outbred white mice weighing 19-22 g at the age of 10-12 weeks at the time of administration of the test compound were used in the experiment. 3-6 mice were selected in the experimental groups without deviations in appearance and with deviations in weight. Each animal was assigned an individual number.

The acute toxicity of compound 1 was studied over a wide range of doses from 100 to 1580 mg / kg with a suspension of C-1 in a 1% starch paste once intraperitoneally. When choosing doses, they were guided by the maximum volume of fluid allowed for intraperitoneal administration to mice. The condition of the mice was evaluated during the first hour, as well as 1, 3 and 7 days after administration. The criteria for assessing acute toxicity were: the number of dead animals, the time of their death, LD ₅₀ after 24-72 hours, a picture of intoxication. Accounting for the death and general condition of animals was carried out once a day daily throughout the entire experiment.

500 мг/кг), что значительно ниже, чем у С-3 (ЛД₅₀=178 мг/кг) [Бурякина А.В. Острая токсичность ронтанокса, обладающего антигельминтной активностью / А.В.Бурякина, Ф.С.Михайлицын, Д.П.Севбо, А.Ю.Малахова, С.Н.Трусов // Российский паразитологический журнал. 2011. №1. С.108-111].The toxicity of the claimed compounds (LD ₅₀

500 mg / kg), which is significantly lower than that of C-3 (LD ₅₀ = 178 mg / kg) [Buryakina A.V. Acute toxicity of rontanox with anthelmintic activity / A.V. Buryakina, F.S. Mikhaylitsyn, D.P. Sevbo, A.Yu. Malakhova, S.N. Trusov // Russian Parasitological Journal. 2011. No1. S.108-111].

Example 3. Evaluation of anthelmintic activity:

Evaluation of the anthelmintic activity of the claimed compound C-1 was carried out on an experimental model of cestodoses - hymenolepidosis (white mice invaded with Hymenolepis nana at a dose of 200 invaded eggs per animal), the compounds were used in micronized form with a particle size of 10-15 μm (obtained by grinding). For each test dose of substances, 8 mice were used; they were administered orally in a starch paste, once on day 13 after infection. The results of the experiment were taken into account one day after treatment according to the number of animals completely freed from invasion (see table).

Research results

The test results of the anthelmintic activity of the claimed compound C-1 during cestodosis in the experimental model of Hymenolepis nana indicate that the drug is highly effective in the studied dose range (see table).

The basic drug - FENASAL does not have anti-cestosis activity at a dose of 250 mg / kg, 125 mg / kg, and the claimed compound 1 (C-1) in the same doses showed 100% effectiveness and showed significant activity at a dose of 0.063 mg / kg. Thus, the results of evaluating the anthelmintic effect of the claimed drug show that C-1 is significantly more effective in anthelmintic action compared with the drug FENASAL. The inventive compound 1 is less toxic compared to C-3, C-4 and more effective compared to C-3, C-4 and C-5.

The specified property allows us to suggest the possibility of using the claimed compounds in medicine and veterinary medicine.

Table Test results of anthelmintic activity (Hymenolepis nana model) of compound C-1 in comparison with C-3, C-4, C-5 and phenasal (comparison drug). Compound Dose g / kg The effectiveness of the treatment -% of animals (mice), completely freed from cestodes Fenasal (C-2) 1,0 one hundred 0.5 62 S-4 0.5 one hundred 0.25 0 S-3 0.5 Death of all animals 0.25 one hundred 0.125 83 Prototype (C-5) 0.5 one hundred 0.25 one hundred 0.125 0 The inventive compound (C-1) 0.25 one hundred 0.125 one hundred 0,063 40.8

Claims (1)

N- (3-Chloro-4-propyloxyphenyl) -3,5-dichlorosalicylamide of the formula 1

possessing anthelmintic activity.