RU2156621C1 - Neurotropic drug - Google Patents

Abstract

FIELD: medicine, pharmacy. SUBSTANCE: neurotropic drug is a potentiated form of antibody-antiserum raised to brain-specific protein S-100. This form is prepared by successive multiple dilution and vertical shaking of solution antibody-antiserum in neutral solvent by homeopathic procedure. Drug stimulates regenerative process without adverse effects effectively. Invention can be used for correction of impaired immune homeostasis accompanying with alternations of psychosomatic functions and/or biological responses by many-sided effect on brain integrative activity. EFFECT: enhanced effectiveness of drug, absence of adverse effects. 9 ex

Description

 The invention relates to medicine and can be used to correct impaired immune homeostasis, accompanied by a change in psychosomatic functions and / or biological reactions, through a multifaceted effect on the integrative activity of the brain.

 The use of antibodies as a medicine for the treatment of various infectious and viral diseases is known (see R.Ya. Meshkova, L.V. Kovalchuk, M.I. Konovalova "Clinic, diagnosis, treatment of certain forms of immunodeficiency and allergic diseases with the basics of organizing a service clinical immunology ", Smolensk," Polygram ", 1995, pp. 56-64; ed. certificate 1131508, class A 61 K 39/00, 1984) The antiviral effect of these drugs based on antibodies specific to the antigenic determinants of a particular virus, strictly choose no, but they called passive immunity lasts a relatively short time.

 Also known is a drug based on antibodies to the pathogen, modified to increase the effectiveness of the immune response with 1-2 fatty acid residues (see ed. St. 1730144, class C 12 N 7/00, 1992). However, such treatment gives the expected effect. not for all antibodies used for therapeutic purposes.

 Closest to the invention can be considered the use of antiserum antibodies to brain-specific proteins as a neurotropic drug for the treatment of certain demyelinating diseases of the central nervous system / cm. book of M. B. Stark, Brain-specific proteins / antigens / and neuron functions, M., "Medicine", 1985; p. 234-236, 224-225 /.

 However, such drugs are characterized by narrow specificity and, as a rule, insufficient activity, which requires the use of additional substances with a stimulating effect, and can cause side effects.

 The invention is directed to the creation of a neurotropic drug based on a potentiated form of an antibody-antiserum to a brain-specific protein that effectively stimulates regenerative processes without side effects.

 The solution to this problem is provided by the fact that a neurotropic drug based on an antibody-antiserum to a brain-specific protein is a potentiated form of an antibody-antiserum to a brain-specific protein S-100, obtained by repeated dilution and shaking according to the homeopathic method.

For the preparation of a potentiated form of an antibody-antiserum, a brain-specific protein S-100 is used, the physicochemical properties of which are described in an article by M.V. Starostina, S.M. Sviridov, Neurospecific protein S-100, Advances in modern biology, 1977, v. 5, p. 170-178; the book of MB Stark, Brain-specific proteins / antigens / and neuron functions, M., "Medicine", 1985; p.12-14, isolated from tissue of the brain of a bull by the following method:
- fabric frozen in liquid nitrogen is ground into powder in a special mill;
- the extraction of proteins is carried out in a ratio of 1: 3 (weight / volume) in the extraction buffer during homogenization;
- the homogenate is heated for 10 min at 60 ^o C and cooled to 4 ^o C in an ice bath;
- thermolabile proteins are removed by centrifugation;
- carry out stepwise fractionation with ammonium sulfate, followed by removal of precipitated impurity proteins;
- the S-100 protein-containing fraction is precipitated with 100% saturated ammonium sulfate when the pH is reduced to 4.0 and collected by centrifugation;
- the precipitate is dissolved in a minimum volume of a buffer containing EDTA and mercaptoethanol, dialyzed against deionized water and freeze-dried;
- fractionation of acidic proteins is continued by chromatography on ion-exchange media — DEAE cellulose DE-52 and then DEAE-Sephadex A-50;
- collected and dialyzed fractions containing S-100 protein are separated by molecular weight by gel filtration on Sephadex G-100;
- purified S-100 protein is dialyzed and freeze-dried.

 The molecular weight of the purified brain-specific protein S-100 is 21,000 D.

 Due to the high content of aspartic and glutamic acids, the brain-specific protein S-100 is strongly acidic and occupies the extreme anode position during electrophoresis in a discontinuous buffer system in a polyacrylamide gel, which facilitates its identification.

 To obtain a brain-specific antiserum, a purified S-100 protein (antigen) is prepared in combination with methylated bovine serum albumin as a carrier with Freund's complete adjuvant, which is administered subcutaneously to a laboratory rabbit animal in the amount of 1- to a selected brain-specific protein S-100. 2 ml On the 8th, 15th day, re-immunization is carried out. Blood sampling is performed (for example, from an ear vein) on the 26th and 28th day.

 The resulting antiserum has a titer of 1: 500 - 1: 1000, forms a single precipitation band with extract from the nervous tissue, but does not react with extracts of heterologous organs, and forms a single precipitation peak with both pure S-100 protein and extract of nervous tissue, which indicates the monospecificity of the obtained antiserum.

 From the obtained antiserum, which is a substance soluble in a neutral solvent (distilled water, physiological saline, alcohol, etc.), a potentiated form is prepared, while the concentration is uniformly reduced by successive dilution of 1 part of the substance in 9 parts (for decimal dilution ) either in 99 parts (for hundredth dilution) or in 999 parts (for thousandth dilution) of a neutral solvent with multiple vertical shaking (“dynamization”) of each dilution and using separate containers for each subsequent dilution to obtain the desired potency - the dilution ratio according to the homeopathic method (see, for example, W. Schwabe "Homeopathic medicines", M., 1967, S. 14-29). A neurotropic drug thus prepared is used in dosage forms and dilutions accepted in homeopathic practice, predominantly hundred (C1-C10000).

Laboratory studies of the biological activity of the claimed neurotropic drug in the form of a potentiated form of an antibody-antiserum to the brain-specific protein S-100 in dilutions of C6-C200 showed the following:
1) in studies on models of the conditional avoidance reaction (URI), the introduction of a potentiated antibody-antiserum form to the S-100 protein for 5 days inhibited the formation of URI regardless of the modality of the conditioned stimulus and unconditional reinforcement (“punishment” or “reward” reaction) ;
2) when studying rat surviving hippocampal slices on a model of long-term post-genetic potentiation (DPTP), it was found that the potentiated antibody-antiserum form to the brain-specific protein S-100 does not affect the potentiation induction; S-100 protein. The modifying effect of potentiated antiserum on synaptic efficacy is due to the manifestation of the phenomenon of systemic adaptation, previously called "bipatic" (see PCT application WO 97/28776, class A 61 J 3/00, A 61 H 39/00, 1997)
3) when studying the effect of potentiated antibody-antiserum to the brain-specific protein S-100 in dilutions C6 and C200 (conventional concentration 10 ^-12 and 10 ^-400 ) on the passive and active characteristics of the membranes of the subcellular ganglion of H.Pomatia, it was found that all variants of potentiated dilutions they have an identical effect with the starting materials, but to a different degree expressed effect: they depolarize the membrane, reduce the amplitude of the action potential and maximum conductivity, and contribute to the growth of channel inactivation.

 The following are examples of the use of a neurotropic drug.

 Example 1

 Boy S., 28 years old. The diagnosis is chronic alcoholism of the II degree, moderate alcohol syndrome. Monotherapy with potentiated antiserum to the brain-specific protein S-100, 5 drops of an alcohol solution in a dilution of C200 5 times a day for 2 days eliminated all manifestations of alcohol withdrawal: nausea, thirst, sweating, headache. Normalized sleep, appetite, mood.

 Example 2. B-naya P., 40 years old. The diagnosis of chronic alcoholism II tbsp. I haven’t taken alcoholic beverages for 3 years, but recently, B-naya has noted a renewed craving for alcohol, a sleep disorder, and a decrease in mood. Reception of a potentiated antiserum antibody to a brain-specific protein in a dilution of C200 in a dose of 1 tablet of 0.25 g in the morning for 2 weeks completely suppressed the desire to drink alcohol, sleep and mood normalized.

 Example 3

 Boy P., 20 years old. The diagnosis of opium addiction, II tbsp. For 2 years, uses increasing doses of heroin, intravenously. Repeatedly treated in hospitals, usually the detoxification course reached 2-3 weeks. However, after discharge from the hospital, he immediately resumed drug use. At the next hospitalization in December 1998, a potentiated drug was included in the complex therapy of heroin withdrawal - a potentiated antiserum to the brain-specific protein S-100, 5 drops of an alcohol solution in a dilution of C100 3 times a day. Withdrawal syndrome is stopped within 7 days. After discharge for 1 month, heroin does not use, when signs of attraction appear, it takes 1 tablet (0.5 g) of the C1000 of the claimed drug - a potentiated antibody-serum to the brain-specific protein S-100.

 Example 4. Patient D., 45 years old, diagnosed with psycho-organic syndrome due to traumatic brain injury, asthenic variant. Disabled person of group II. In spring and autumn, the condition worsens: headaches, irritability, meteorolability increase, sleep is disturbed. The patient quickly gets tired, with difficulty caring for himself. The last period of exacerbation for 2 months took only the claimed neurotropic drug in a dilution of C500, 1 tablet of 0.25 g once a day. His condition improved, the patient was more active, his sleep and mood normalized. No headaches. He is engaged in physical therapy. Subjectively assesses his condition as satisfactory.

 Example 5

 B-naya K., 43 years old. Is in a psychiatric hospital with a diagnosis of endogenous depression of unspecified origin. Conducted for 3 weeks, antidepressant and tranquilizer therapy is ineffective, complains of lethargy, insomnia, and "insensibility." After the potentiated antibody-antiserum to the brain-specific protein S-100 was added to the therapeutic doses of antidepressants in a dilution of C1000, 5 drops of an alcohol solution 8 times a day, the mood was leveled for 10 days. Discharged in satisfactory condition.

 Example 6

 B. Naya, 32 years old. The diagnosis is neurasthenia. Complains of weakness, fatigue, headache, irritability. On an outpatient basis, I took in the morning 1 tablet of C200 of the claimed neurotropic drug - a potentiated antibody to the brain-specific protein S-100. The state within 5 days has changed dramatically: the former is stenic, active, mood, sleep, no irritability.

 Example 7

 Boy P., 19 years old, student. Complains of fatigue, inability to concentrate, academic performance has declined, she is afraid not to pass the next session. An effective course of nootropil in such cases did not bring an increase in efficiency. Reception during the week of the claimed neurotropic drug 1 tablet (0.25 g) 2 times a day of a potentiated antibody-antiserum to the brain-specific protein S-100 in a dilution of C50 quickly restored performance, increased attention. Bboy successfully passed the exams.

 Example 8

 B-naya T., 80 years old. Lives with a disabled daughter. Due to memory impairments, she can’t cope with homework, forgets the way to the store. The course of treatment for 6 months with the claimed neurotropic drug of 5 drops of alcohol solution C200 led to a noticeable improvement in memory. G-naya began to care for her daughter, walks the streets, does homework.

 Example 9

 Boy S., 49 years old. Diagnosis of inoperable cerebellar neuroblastoma. Sick for 3 years, underwent a course of chemo- and radiation therapy. Despite treatment, the condition is serious: due to impaired gait and weakness, it is not capable of independent movement, cachexia is increasing. Prescribed: potentiated antiserum to the brain-specific protein S-100B, which is one of the fractions of the brain-specific protein S-100, in the dilution of C1000 5 drops of an alcohol solution 2 times a day. Against the background of therapy, after 4 months, the patient’s condition stabilized somewhat: gait disturbances decreased (he began to move with a stick) and weakness, appetite appeared. The condition has stabilized, the tumor does not grow.

Claims (1)

 A neurotropic drug based on an antiserum to a brain-specific protein, characterized in that it is a potentiated form of an antiserum in the form of a solution of antibodies, an antiserum to a brain-specific protein S-100 in a neutral solvent, obtained by successive repeated dilution and vertical shaking.