RU2136303C1 - Method of preparing agent "cheblin sk-1" for treatment of larval and strobillar echinococcosis - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: walnut fruits at the stage of milk ripeness are extracted with kerosene at ratio raw : extractant = (1:0.8)- -(1:1.5). EFFECT: enhanced activity, decreased toxicity of an agent. 5 ex

Description

 The invention relates to medicine, in particular to agents for treating echinococcosis.

 To suppress the development of larval echinococcosis in animals, medamine has been proposed [1].

 The disadvantage of the drug is that after the course of treatment there are viable elements of echinococcus, which after the cessation of treatment begin to develop again. In addition, the maximum non-lethal dose (MNLD) of medamine is 2.5 g / kg, the Cheblin SK-1 preparation we offer is less toxic, the MNLD for white outbred mice of our preparation is 19 g / kg.

 Known drug G-1697/2 /, which is 4 - [(benzo-2,1,3-thiadiazolin-4) amino] -5,6,7, non-lethal dose / MNLD /, established by the authors in the experiment on white outbred mice was 4 g / kg

 The disadvantage of the drug / 2 / is a higher toxicity compared with our drug.

 The objective of the invention is the search for herbal remedies with high anti-echinococcosis activity and lower toxicity compared to known drugs.

 The task is achieved by obtaining funds Cheblin - SK-1 by extraction with purified kerosene fruits of walnut milk ripeness.

 The method is as follows.

Example 1. 100 kg of walnut fruits in the stage of milk maturity are crushed, the resulting mass is poured into 100 kg of purified aviation kerosene. Maintain for 40 days in a dark place at room temperature not higher than 21 ^o C with periodic stirring. The resulting mixture is filtered, the liquid part is squeezed out of the remaining part, which is again filtered, the first and second portion of the extract are combined. Then, spectrophotometric, chromatographic analyzes are carried out, which provide quantitative determination of carotene, chlorophyll, and nut oil.

 Example 2. 100 kg of walnut fruits in the stage of milk maturity are crushed, the resulting mass is poured into 150 kg of kerosene for medical use. Stand for 40 days at room temperature in a dark place. The resulting mixture is filtered, the liquid part is squeezed out of the remaining part, which is again filtered, the first and second portion of the extract are combined.

 Example 3. 100 kg of walnut fruits in the stage of milk maturity are crushed, the resulting mass is poured 80 kg of kerosene for medical use. Stand for 40 days at room temperature in a dark place. The resulting mixture is filtered, the liquid part is squeezed out of the remaining part, which is again filtered, the first and second portion of the extract are combined.

 Raw materials are procured in May, washed from dirt, dust, impurities, separated from the stalk, ventilated in air and finely chopped.

 Cheblin SK-1 was effective in strobilar echinococcosis of dogs and echinococcosis of piglets. A three-week course of treatment is also offered for people with echinococcosis. With strobiliary echinococcosis, a single dose of Cheblin-SK-1 was 1 g / kg, with rat alveolar echinococcosis, hydatidosis echinococcosis of piglets and humans, the first week of treatment was 0.1 g / kg for rats and 0.05 g / kg for piglets and people, the second week of treatment is 0.26 g / kg and 0.1 g / kg, the third week is 0.34 g / kg and 0.25 g / kg.

 Anti-echinococcosis (anthelmintic) efficacy was established in the experiment, 22 dogs infected with strobiliary echinococcosis confirmed by special helminthological studies were divided into two groups: 8 control, 14 received 4 g / kg of Cheblin SK-1 preparation inside. 5 days after giving the drug, the dogs were opened, their intestines were examined by the method of complete helminthological dissections according to K.I.Skryabin. In the intestines of control dogs, from 64 to 620 specimens of echinococci were found (all infected) on average 274.8 ± 69.8 specimens of mature echinococci. Anthelmintic effectiveness reached 100%.

 Cheblin SK-1 showed high anthelmintic efficacy in experimental alveolar echinococcosis of cotton rats. 21 cotton rats were used in the experiment. Animals are infected by a known method [3, 4]. By the start of treatment, the duration of experimental invasion was 10 days, when in 5 opened rats the mass of parasitic larvocysts ranged from 0.09 to 0.49 g and averaged 0.29 ± 0.07 g. Subsequently, 8 infected rats received the Cheblin-SK-1 preparation for three weeks in gradually increasing daily doses.

 Treated and control rats were euthanized 40 days after infection. Studies and calculations were carried out by a known method [3]. The growth inhibition indices of parasitic larvocysts were ITRL-1 = 90.23%. ITRL-2 = 92.15%, ITRL-3 = 92.74. Destruction of all germ elements was detected in 75-95% of parasitic larvocysts among all detected in each treated animal. In control rats, all germ elements are viable morphologically unchanged.

 With hydatidosis echinococcosis, the activity of the SK-1 preparation was established on 26 piglets, divided into 4 groups: the first and fourth, 5, the second third - 8 animals each. At the age of one month, piglets of groups 1, 2, and 3 were infected with protoscolexes and acephalocysts of a single-chamber echinococcus from an operated patient. 5000 protoscolexes and acephalocysts (taken together) from a single-chamber human echinococcus were injected intraperitoneally to each piglet. Piglets of the fourth group in the experiment were not infected and did not receive the drug Cheblin SK-1.

 On the 60th day, the pigs of the first group were killed in order to determine the initial mass of parasitic larvocysts (PL) developed by that time, which varied from 1.8 to 9.7 g and averaged 6.3 ± 1.4 g. Piglets of 3 groups, starting from the 61st day from the moment of infection, received Cheblin - SK-1 preparation with food for three weeks in gradually increasing daily doses. The average daily dose during the entire course of treatment was 0.31 g / kg, and the total course dose of the drug reached an average of 2.8 g / kg. 8 piglets of the second group made up the control group - the drug was not received.

 On 91 days after infection, the animals were opened. All pigs of groups 2 and 3 were infected with echinococcus. Animal studies were conducted according to the methods of Mikhailichin F.S. et al. [3]. The mass of the submarine. per animal in untreated animals was 164.60 ± 22.60 g, and in those treated 15.00 ± 0.03 g (the difference between the compared values was statistically highly reliable, P. <0.01). In treated animals, pronounced destruction of the germinal elements was detected in 75-85% of parasitic larvocysts. In untreated animals, larvocysts were well developed, and protoscolexes were viable, without manifestations of destruction.

 The introduction of the drug Cheblin SK-1 for pigs for three weeks did not cause side effects and toxicity, on the contrary, under the influence of our drug normalization of growth and development of piglets, normalization of the total blood count, serum protein and protein fractions, as well as free amino acids.

 The toxicity of the drug was studied on outbred 142 mice and 82 rats, on 78 chickens of 3 months of age Ala Tau breed and 30 chickens of breed Heisigs brown, on 30 piglets of one month old age, on 14 dogs of 5 months of age. The embryotoxicity and teratogenicity of the drug was studied in 190 nine-day-old chicken embryos, in 6 pregnant pigs with 27 offspring obtained from them.

 It was found that the MLND of the Cheblin SK-1 preparation was 19 g / kg for white mice and 22 g / kg for white rats. Compared with white mice and rats, chickens are more sensitive to the drug: LD 50 = 14.0 ± 1.4 g / kg, at P = 0.05 t = 2.06. The drug does not have a teratogenic property. The drug, introduced into the yolk sac to 9-day-old chicken embryos in doses of 0.5-5.0 - 50.0 mg / kg, did not show embryotoxicity and teratogenicity. Long-term administration to experimental animals two, three times higher than the course dose of the drug did not cause toxic and side effects, which is confirmed by histopathological and pathomorphological and hematological studies.

 The drug Cheblin SK-1 is low toxic when administered intragastrically to mice, rats, chickens, dogs, piglets. The therapeutic index is over 350.

In the surgical clinics of the Samarkand State Medical Institute. I.P. Pavlova, the drug was tested on volunteers. 186 patients with echinococcosis with different localization, who before the operation took Cheblin SK-1, were admitted to the test. As a control, there were 26 patients with echinococcosis who did not take the drug. Before the start of treatment and at the end of the course, all patients underwent special studies along with generally accepted ones:
1. The study of blood serum by the reaction of scolexoprecipitation, the formulation of the reaction of latexagglutination and indirect hemagglutination.

 2. A study of peripheral blood, including the determination of the number of red blood cells, hemoglobin, white blood cells, white blood cell count and ESR.

 3. Biochemical studies: total protein, thymol test, bilirubin, urea, glucose, seromucoids, hydroxyproline, protein fractions of blood serum, free amino acids of blood serum and urine.

 4. Immunological studies: T-lymphocytes, killers, - suppressors, immune complexes, anti-idiotypes (ACE), immune leukocytes, neutrophils, eosinophils, lymphocytes, monocytes.

 After the course of treatment of patients with the drug Cheblin SK-1, a thorough study of parasitic larvocysts of echinococcus, extracted by surgeons during the operation, was conducted. In treated patients, 85-95 cases of destruction were observed with the death of the embryonic elements of parasitic larvocysts in 26 patients (control group) who did not receive the drug, but operated on for echinococcosis, larvocysts contained viable, morphologically unchanged germinal elements.

Patients transferred the prescription of the drug without complaints and clinical complications. It was established that the drug was tolerated by patients without adverse reactions, has a beneficial effect on patients, restores the protein-amino acid composition in patients with echinococcosis, the immune status improves, which was not observed in control patients
In 17 volunteers with echinococcosis, the anti-echinococcosis effect of Cheblin-SK-1 was studied in increasing doses in a three-week course.

 Example 1. Patient K., 29 years old. Diagnosis: echinococcosis of the brain and left lung. The duration of the disease is 2 years. The clinical picture is dominated by complaints of headaches, nausea, loss of appetite, impaired coordination of movement.

 The reaction of scolexoprecipitation (RSCP), latex agglutination, and indirect hemagglutination are positive.

 Computed tomography of the head revealed a ring-shaped, with clear boundaries, zone of increased density in the region of the pole of the left frontal lobe of the brain with a diameter of 14 mm, a reduced density inside the pole, a similar formation is symmetrically located in the right frontal lobe of the brain with a diameter of 12 mm In addition, 4 spherical formations with a density close to the density of water with a diameter of 4-5 mm were found in various parts of the brain.

 On a radiograph in the upper lobe of the left lung, an echinococcus of spherical shape, with a diameter of about 30 mm, is clearly visible.

 The patient underwent a three-week treatment with Cheblin SK-1. The patient drug was well tolerated, she only felt an unpleasant odor. After the course of treatment, the general condition of the patient began to improve, nausea and headaches disappeared, coordination of movement was restored.

 After 6 months, the course of treatment was repeated. The general condition of the patient has improved dramatically. A year after the second course of treatment, the patient gave birth to a girl weighing 3600 g, the girl grows and develops normally.

 Two years after the second course of treatment, computed tomography of the brain and radiography of the lung were done. On the tomogram in the places where the echinococcus is detected, calcified areas, on the roentgenogram in the lung, the calcified echinococcus was asleep.

 Immunological reactions: RSKP, latexagglutination, indirect hemagglutination negative.

 Example 2. Patient M. 36 years. Diagnosis: multiple recurrent pulmonary echinococcosis. The duration of the disease is 6 years. Immunological reactions: RSKP, latexagglutination, indirect hemagglutination positive.

 The patient was first operated on for echinococcosis of the spleen, during the operation there was a rupture of 7 × 8 cm larvocysts. Four years later, the patient was operated on for liver echinococcosis. A year after the operation, a multiple echinococcal lesion of the lungs was revealed on the radiograph: in the left lung there were 14 echinococci 6-8 mm in diameter, in the right lung 11 echinococci of the same size. The patient is demoralized. A course of psychotherapy was conducted, a meeting was organized with patients who recovered. The patient arbitrarily doubled the dose of the drug. The patient endured the course of treatment well. The indicators of the general analysis of blood and urine remained at the initial level. Biochemical parameters of blood and urine also remained unchanged. After 6 months, the second course of treatment was carried out (double dose of the drug). The treatment went without complications. The general health of the patient began to improve.

 Two years after the second course of treatment, control studies were performed, which calcified areas were found on the radiograph of the lung. Immunological reactions: RSKP, latexagglutination, indirect hemagglutination, negative. The patient recovered.

 Based on the tests, it was found that the drug Cheblin SK-1 is easily tolerated by patients, without adverse reactions, has a beneficial effect on patients with echinococcosis, first there is inhibition of the growth of parasitic larvocysts with destruction and death of 85-95% of the germinal elements of the larvocyst with subsequent death of the parasites. After a course of treatment in patients with echinococcosis, protein-amino acid metabolism is restored.

List of references
1. Kovalenko F.P., Pereverzeva E.I., Dzhabarova V.I. et al. / RF Patent N 1813442 published in 17 bullets. for 1993 g. Means for suppressing larval echinococcosis in animals.

 2. Mikhailicin F.S., Kovalenko F.P., Kozyreva N.P. and other. The search for new antiparasitic drugs. 17. Synthesis and study of anti-echinococcosis activity of the new drug G-1697 // Honey. parasitology and parasitic diseases. - 1996. - N 3 - S. 38-42.

Claims (1)

 A method of obtaining funds for the treatment of larval and strobiliary echinococcosis, which consists in extracting walnut fruits with purified kerosene in the stage of milk ripeness with a ratio of raw materials: extractant 1: 0.8 - 1: 1.5.