RU2550948C1 - N,n'-(sulphonyldi-p-phenylene)bis-n",n"- dimethylformamidine, possessing acaricidal action - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: as the substance, possessing the arcaricidal action, applied is N,N'-(sulphonyldi-p-phenylene)bis-N",N"-dimethylformamidine.

EFFECT: application of the claimed substance assists to deal with ticks and mites in an efficient and safe way.

6 dwg, 8 tbl, 6 ex

Description

FIELD OF THE INVENTION

The invention relates to the field of veterinary medicine, medicine and agriculture, in particular to the use of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine as a substance with acaricidal action.

State of the art

The arthropod subclass Acarina is extremely widespread in nature. Many of them parasitize on various species of animals and plants, and also cause various diseases in humans. Some types of ticks not only harm the health of animals and humans as a result of parasitic nutrition (blood), but also suffer many extremely dangerous, often fatal diseases, infecting animals and humans by suction. In the first half of the 20th century, quite active acaricides were proposed - polychlorinated insecticides, the tolerance of which slowly developed in arthropods of the Acarina subclass. This made it possible to significantly control the number of these parasites. However, later on, very dangerous environmental consequences of the use of insecticides of this class were discovered - this is due to their very high stability in the natural environment, in the body of animals and their accumulation in food chains, which led to the threat of the disappearance of whole animal species. In addition, the production and use of most polychlorinated insecticides pollutes the environment with polychlorinated dioxins, which have very high immunotoxicity and are also very stable. As a result, the use of acaricidal insecticides of this group by the end of the 20th century was universally banned or significantly limited. Instead of polychlorinated insecticides, synthetic analogues of natural insecticides, pyrethroids, which showed a significant acaricidal effect, were introduced into practice, but it was soon discovered that arthropods of the Acarina subclass very quickly become tolerant to this type of insecticide, almost any pyrethroid, both natural and synthetic. Although very effective insecticides of new groups were developed by the beginning of the 21st century — chitin synthesis inhibitors, neonicotinoids, juvenile hormones, pyrazolones, avermectins, which are highly active with respect to arthropods of the class Insecta, they were either inactive with respect to arthropods of the Acarina subclass or ticks tolerance quickly developed and they lost sensitivity to all drugs in the group.

In the second half of the 20th century, high acaricidal activity of dimethylformamidine derivatives of arylamines was discovered, of which formate (3 - [(EZ) -dimethylaminomethyleneamino] phenylmethylcarbamate, cas No. 22259-30-9) and chlordimeform (N ′ - (4- chloro-2-methylphenyl) -N, N-dimethylformamidine, cas No. 6164-98-3). However, formethanate was extremely toxic to warm-blooded rats, LD50 (yelling) = 20 mg / kg [World Review of Pest Control. - 1970. - Vol. 9. - P. 119.]. Chlordimeform, although it has less toxicity, rats LD50 (yelling) = 160 mg / kg [Chemistry in agriculture. - 1978. - No. 2. - from. 59.], but is not safe, because it turned out that the products of its decomposition are nephrotoxic and very stable, accumulate in soil and plants, which is environmentally unacceptable [N.N. Melnikov, A.I. Volkov, O.A. Korotkova. Pesticides and the environment - M .: Chemistry, 1977. - 240 s]. Both products have been virtually discontinued. The most successful drug of the formamidine group was N, N-di (2,4-xylyliminomethyl) methylamine (cas No. 33089-61-1) (hereinafter referred to as amitraz), which is now widely used in veterinary medicine and agriculture. However, this drug also has several disadvantages:

- Quite high toxicity, rats LD50 (oral) = 400 mg / kg [Merck Index - 11th ed. - 1989. - Vol. 11. - P. 80], which excludes the use in medicine and complicates its use in veterinary medicine.

- Low chemical resistance, amitraz is easily hydrolyzed and oxidized with the loss of acaricidal activity, which also complicates its use.

- Amitraz exhibits toxicity due to the effects of α ₂ -adrenergic agonism and inhibition of monoamine oxidase in animal brain tissue. This leads already at doses of about 1 mg / kg of the animal’s weight to develop behavioral disorders, and at doses of about 100 mg / kg, intoxication becomes life threatening [J. Rouder. Veterinary toxicology. - M. Aquarium, 2008. - 416 s].

Disclosure of invention

The objective of the invention was to search for a substance of the formamidine group that has high acaricidal activity at low toxicity for warm-blooded and high chemical resistance, including in aquatic environments, but at the same time easily destroyed in the environment without the formation of hazardous products. Of the arylamine derivatives we studied with formamidines, the best in all parameters (high acaricidal activity, low toxicity) was Ν, Ν ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine (Methanimidamide, N ′, N ″ ″) - (sulfonyldi-4,1-phenylene) bis [N, N-dimethyl-, cas No. 3217-65-0).

N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine has long been known [A.W. Hofmann. Annals. - 1858. - Vol. 54. - P. 197]. It was studied as an antibacterial, antiprotozoal, antiviral and anthelmintic agent by Steiger [US 3073851, publ.: 01/15/1963] and on immunomodulating and antitumor activity by Lin [EP 354303, publ.: 02/14/1990].

The methyluracil salt of N, N ′ - (sulfonyldi-p-phenylene) bis-N, N-dimethylformamidine (hereinafter referred to as sulphiphone) was proposed as stimulating cell metabolism, exhibiting high immunotropic and antimycobacterial activity [RU 2136668, publ.: 09/10/19991, however, it was not used in medicine and veterinary medicine.

Also proposed is an immunomodulating agent containing sulphiphone and organic acid (ascorbic, succinic or citric) [RU 2292892, publ.: 02/10/2007], which was also not accepted for use in medicine and veterinary medicine.

The extremely high broad-spectrum acaricidal activity of Ν, Ν ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine was unexpected. In the available literature, no indications of this activity of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine were found by us. The proposed compound with high efficiency cures invasive pathologies in animals caused by scabies mites (psoroptosis, sarcoptosis, chorioptosis, etc.), while the clinical effect is significantly more pronounced than when using amitraz (see example 4). With a high degree of probability, it can be assumed that an identical effect will be observed with invasions in humans (sarcoptosis, notohedrosis). N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine proposed as an acaricide also destroys ixodid ticks - dangerous carriers of viral diseases, pathogenic rickettsiae and protozoa (see example 3; 5). The compound has low toxicity, rats LD50 (oral) = 4750 mg / kg (see example 1), this makes it safe to use in veterinary medicine / medicine and makes it possible to use for systemic administration (oral, injectable, transdermal). The compound also exhibits an anti-inflammatory effect (see example 2), reducing the level of inflammatory mediators in all groups; this may also increase the clinical effect, accelerating the healing of damaged animal tissue during tick-borne infestations (see example 4).

In comparison with the widely used amaritraz acaricide, the proposed compound showed significantly greater activity against scabies, ixodidae, phytopathogenic ticks (see examples 3; 4; 5; 6).

In addition, the proposed compound is more stable in aqueous media than amitraz (see example 6). In the environment, N, N ′ - (sulfonyldi-p-phenylene) bis Ν ″, Ν ″ -dimethylformamidine decomposes to the known 4,4′-diaminodiphenylsulfone (dapsone), a low-toxic compound, rat LD50 (oral) = 1000 mg / kg [Logemann W, Miori GP Arzneimittelforschung. - 1955. - Vol. 5. - P. 213-221].

Amidine acaricides in the environment quickly hydrolyze to formamides of the corresponding anilines and then relatively slowly to free aniline. The latter undergoes various transformations depending on the structure. Chlordimiform forms 2-methyl-4-chloraniline, which is stable in the environment and is able to accumulate in plants, while at the same time it possesses nephrotoxicity like all p-chloranilines. Amitraz forms 2,4-xylidine, which is relatively quickly oxidized at the methyl group in the para position and is assimilated by soil microorganisms as an analogue of p-aminobenzoic acid. However, 2,4-xylidine is quite toxic to warm-blooded rats, LD50 (oral) = 467 mg / kg [National Technical Information Service. PB 214-270].

N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine slowly hydrolyzes as a result to 4,4′-diaminodiphenylsulfone (dapsone), which is also a structural analogue of p-aminobenzoic acid and is assimilated by soil microorganisms which eventually oxidize the structure to carbon dioxide and ammonium sulfates. Due to the rapid assimilation of decomposition products of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine by microorganisms in plants, they do not accumulate and do not fall into food chains (unlike decomposition products of chlordimeform).

The structure of the aromatic fragment of the molecule N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine excludes binding to α ₂ -adrenoreceptors. N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine does not affect the activity of monoamine oxidases, because the acceptor effect of the arylsulfonyl group reduces the basicity of the formamidine structure to a level that excludes binding to the enzyme. When studying toxicity, no effect of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine on the central nervous system functions of animals was found.

Thus, a substance was found (N, N ′ - (sulfonyldi-p-phenylene) bis-N, N ″ -dimethylformamidine), which has a wide spectrum of acaricidal action, low toxicity, chemically stable, including in aqueous media, and decomposes under the action of natural factors without the formation of hazardous products. Based on the proposed N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine, acaricidal preparations for use in medicine, veterinary medicine, agriculture, household chemicals, etc. can be obtained by methods well known to specialists. d.

Brief Description of the Drawings

In FIG. 1 shows a chorioptotic colony before gel treatment with Ν, Ν ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine gel.

In FIG. 2 shows the results of gel treatment based on N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine after 7 days.

In FIG. 3 shows the results of gel treatment based on N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine after 14 days.

In FIG. 4 shows a chorioptotic colony prior to amitrase-based gel treatment.

In FIG. 5 shows the results of gel treatment based on amitrase after 7 days.

In FIG. Figure 6 shows the results of gel treatment based on amitrase after 14 days.

The implementation of the invention

Example 1. (Acute toxicity of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N′-dimethylformamidine)

Materials and methods

The acute toxicity of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine was studied in white rats weighing 200-210 g with intragastric administration in the form of an aqueous solution of the drug in increasing doses. In the experiment, clinically healthy animals were used, which were previously kept in quarantine for 15 days. Statistical groups consisted of 6 animals.

results

It was found that LD50 (v / f) = 4750 mg / kg, which allows this drug to be classified as hazard class 3 (moderately hazardous substances) according to GOST 12.1.007-76.

The results of determining the parameters of the acute toxic effect of the drug with the intragastric route of administration are presented in Table 1.

The clinic of acute animal poisoning was characterized by severe depression, lethargy, and decreased mobility. Death occurred as a result of respiratory arrest.

Example 2. (Evaluation of the anti-inflammatory and immunomodulatory activity of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine)

Anti-inflammatory and immunomodulatory activity of the drug was evaluated by standardized biochemical and hematological methods according to the content of ESR, CRH, sialic acids, fibrinogen, leukocytes and “cytokine status”.

Materials and methods

In the experiment, male rats weighing 180-200 g were used. Each group included 10 individuals. Chronic immune inflammation was modeled by subplanar injection of 0.1 ml of Freund's adjuvant into the right hind paw (BCG suspension 2.5 mg / ml in liquid paraffin). The test drug was started intragastrically from 5 days after adjuvant administration daily for 12 days. In total, 4 experimental groups were investigated:

group - intact + starch suspension;

group - adjuvant + starch suspension (arthritis without treatment);

3 group - adjuvant + N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine (20 mg / kg in starch suspension);

Group 4 - N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine (20 mg / kg in starch suspension). Evaluation was carried out on 10, 16, 19 and 22 days after administration of the adjuvant.

findings

The data obtained indicate the presence of a pronounced anti-inflammatory (see Table 2) and certain immunomodulatory properties in the test preparation under conditions of inflammation (see Table 3). Administration of the drug to intact animals does not affect their “cytokine status”.

Example 3. (Study of the acaricidal activity of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine, the effect on ticks)

Materials and methods

Two acaricidal preparations of different composition were compared.

Composition 1, wt.%:

Isopropyl alcohol thirty Polyvinylpyrrolidone 40 Twin 80 5 N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ - 0.31 Dimethylformamidine Polyethylene glycol up to 100

Composition 2, wt.%:

Isopropyl alcohol thirty Polyvinylpyrrolidone 40 Twin 80 5 Amitraz 0.31 Polyethylene glycol up to 100

The tests were held in the laboratory of parasitology GNU SNIIZHK. The experiments (in vitro) were carried out on isolated ticks Dermacentor pictus, Dermacentor marginatus, Hyalomma plumbeum.

A) 12 × 12 cm tissue wipes were treated with experimental preparations by spraying. Soaked for 1 hour. The processed napkins were placed at the bottom of the Petri dish, where 5 copies of hungry individuals (males and females) Dermacentor pictus, Dermacentor marginatus were planted. Similarly, they acted with control (n = 5), placing ticks on an untreated napkin. The same manipulations were done with sheared dog hair. Acaricidal effect was checked by observation after 1, 4, 6, 24, 48, 72 hours, taking into account the number of dead, paralyzed and living ticks.

B) The experiment was carried out similarly to experiment A, but wipes and wool were not kept for 1 hour before planting ticks. Five copies of hungry individuals (males and females) of Hyalomma plumbeum were planted in each cup. They did the same with control (n = 5), placing ticks on untreated material. Acaricidal action was checked by observation after 5.10 minutes, 1, 6, 24, 48 hours, taking into account the number of dead, paralyzed and living ticks.

findings

A) N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine has acaricidal activity. According to the results of studies, when processing with composition 1 (N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine), mite death occurred in 80% of cases, and when processing with composition 2 (amitraz) - in 60% cases (see Table 4).

B) The ticks placed in the centers of the Petri dishes instinctively began to look for a shadow in the wool, which did not scare them away, they buried and crawled over it, indicating a low repellent effect of the studied drugs. Ticks placed on wipes already lost activity during the first 2 minutes. In both cases, the death of all experimental subjects occurred in 10-15 minutes (see Table 5).

Example 4. (Study of acaricidal activity of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine, effect on scabies mites)

Materials and methods

The experiments were carried out on the basis of the SEC "Kobralovsky" (Leningrad region, Gatchinsky district). For the experiments, 14 cows were selected at the age of 3 to 7 years with clinical signs of chorioptosis. The disease was manifested by a scab in the area of the root of the tail and the udder of the mirror. Chorioptotic colonies reached 15 cm in length (see Fig. 1; 4). In scrapings taken from the lesion sites, Chorioptes bovis ticks were found at all stages of development.

Investigated 2 types of gel:

Composition 1, wt.%:

N, N ′ - (sulfonyldi-p-phenylene) bis- 5 N ″, N ″ dimethylformamidine Complex 3- (2,2,2- 10 trimethylhydrazinium) propionate-2-ethyl- 6-methyl-3-hydroxypyridine disuccinate Chitosan Succinamide 10 Polyoxyethylene tocopherol one Water for injections up to 100

Composition 2, wt.%:

Amitraz 5 Complex 3- (2,2,2- 10 trimethylhydrazinium) propionate-2-ethyl- 6-methyl-3-hydroxypyridine disuccinate Chitosan Succinamide 10 Polyoxyethylene tocopherol one Water for injections up to 100

The animals were divided into 2 groups:

group - gel treatment of Composition 1 (7 animals);

group - gel treatment of Composition 2 (7 animals).

Treatments were carried out once a week for two weeks by rubbing gels on the affected areas of the skin. The result was taken into account 7 and 14 days after the first treatment visually and by taking a scraping.

findings

It was found that skin cleansing during gel treatment based on N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine occurs more intensively than gel treatment based on amitrase (see Table 6).

Example 5. (A study of the acaricidal efficacy of an ethanolic solution of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine on ticks)

Materials and methods

Test Solutions

Solution 1 - 0.01% solution of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N′-dimethylformamidine in 95% ethanol

Solution 2 - 0.01% solution of amitrase in 95% ethanol

Solution 1a - 0.1% solution of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine in 95% ethanol

Solution 2a - 0.1% solution of amitraz in 95% ethanol

The experiment (in vitro) was carried out on isolated ticks Dermacentor pictus, Dermacentor marginatus, Hyalomma plumbeum. For this, 10 copies of hungry adults were placed on fabric (calico or satin) napkins 12 × 12 cm in size, tied with a thread and immersed for 1 min in the test solution. Then they were placed in Petri dishes and in a thermostat at a temperature of 27-30 ° C and a humidity of 95%. They did the same with control, immersing the same number of ticks in 95% ethanol.

Acaricidal effect was checked by observation after 1, 4, 6, 24, 48, 72 hours, taking into account the number of dead, paralyzed and living.

findings

All experimental ticks already lost the ability to move after an hour, died in a day, which indicates a high acaricidal effect of the studied drugs. A 0.1% solution of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine in ethanol has a greater acaricidal effect than a similar solution of amitraz (when using amitraz solution, one of 10 ticks retained active life) ( see Table 7).

Example 6. (Study of the action of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine on phytopathogenic ticks)

Materials and methods

Three plants of Fatsia japonica of approximately the same size, heavily infested with Tetranychus cinnabarinus ticks, were treated. Plant leaves were pale yellow with necrotic spots on the lower side coated with a continuous layer web, wherein the actively moved mites per 1 cm ² were 5 to 20 individuals. As acaricides, 2% alcohol solutions of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine, amitrase and malathion were used, which were diluted with water to a concentration of 0.1% before treatment. Each plant was treated with an aerosol dispenser (100 ml of the test solution per plant). The condition of the plants was evaluated after 1 day, 14 days, 45 days after treatment.

The results are presented in Table 8.

findings

N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine provided effective and long-term protection of plants from phytopathogenic ticks, its phytotoxicity was not detected.

The moderate effect of amitrase, most likely, is associated with rapid decontamination in a humid environment, including on the surface of the leaves. The inventive compound N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine under these conditions is stable and provides a long-term protective effect (after 45 days the acaricidal effect did not stop).

Claims (1)

The use of N, N ′ - (sulfonyldi-p-phenylene) bis-N ″, N ″ -dimethylformamidine as a substance with acaricidal action.

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