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Own research
A scientific study conducted at the E.D. Goldberg Research Institute of Fipm of the Tomsk NIMC
The aim of the study is to develop a new drug with a highly effective effect aimed at combating Alzheimer's disease.
Materials and methods of research
The object of the study was the amount of polyprenols isolated from Siberian Fir (Abies sibirica Ledeb.), the Pine family (fam. Pinaceae).
The experiments were performed on 108 outbred sexually mature male stock mice with an average body weight of 28-30 g (at the age of 5 weeks) conventional, category 1, obtained from the Department of Experimental Biomodeling of the E.D. Goldberg Research Institute of Tomsk NIMC.
The design of the experiments and the maintenance of the animals were approved by the Ethics Committee of the E.D. Goldberg Research Institute of the Tomsk NIMC.
The antiamnestic activity of the substance - the sum of polyprenols was studied in the dose range 5, 20, 50, 100, 200, 500 mg/kg. An experimental model of Alzheimer's disease was created by intraperitoneal administration of scopolamine at a dose of 2 mg/kg [11] for 20 days. Choline alfoscerate (Gliatilin – Catalent Italy S.P.A.) was used as a comparison drug.
All the animals were divided into 9 groups of 12 individuals each. The first group (intact control) was injected intraperitoneally with saline throughout the experiment. The remaining groups were also injected intraperitoneally with scopolamine for 20 days, and from 21 to 30 days intraperitoneally with saline. In addition to saline, 0.4 ml of water was injected into the stomach of the second group (negative control). The third group received the comparison drug gliatilin at a dose of 90 g/kg from day 21 to 30 (positive control) [12]. Groups from the fourth to the ninth received the claimed remedy according to the indicated doses. On the 31st day of the experiment, 1 hour after the introduction of substances, a conditioned passive avoidance reflex (URPI) was developed in all animals. The reflex was checked 24 hours and 7 days after production.
The technique of the conditioned reflex of passive avoidance (URPI) is based on the suppression of the innate reflex of preference for dark space available in rodents [10]. The experimental setup consisted of a camera consisting of two compartments: a large one – illuminated and a small one - dark. The animal was placed in a light compartment and soon (after 10-20 seconds), due to the innate reflex of preference for dark space, it moved into a small compartment, after which the door connecting both compartments was blocked and an electric current was applied to the floor of the dark compartment, consisting of parallel alternating electrodes, with pulses of 50 Ms duration, frequency 5 Hz and amplitude 50 mA. After 10 seconds, the door was opened and the animal could jump out into a light compartment with an ordinary floor. As a result of the described procedure, the animals developed a conditioned reflex of avoiding dark space. When checking the reproducibility of the reflex, the animals were placed in a light compartment, in the corner opposite from the entrance to the dark compartment and observed for 3 minutes. The time of the first entry into the dark compartment (latent entry time) and the total time spent in the dark compartment were recorded.
Statistical processing of the obtained results was carried out using the software package "Statistica 6.0" for Windows [13].
Results and their discussion
In the course of the conducted studies, it was found that scopolamine administered once at a dose of 2 mg / kg for 20 days caused a distinct violation of the production and reproducibility of URPI. At the same time, the course administration of the substance - the sum of polyprenols for 5 days had an improving effect on the reproducibility of the reflex exceeding the effect of the comparison drug Gliatilin. A decrease in the effect of a substance in large doses, versus its effect in small doses, is a phenomenon quite common in the pharmacology of receptor drugs (see Table 1).
Table 1 – The effect of the course administration of the sum of polyprenols on the production of URPI according to the results of checking the reproducibility of the reflex 24 hours and 7 days after production in mice with impaired cognitive and amnestic processes caused by chronic (20 days) administration of scopolamine at a dose of 2 mg/kg (X±m; n=12)
A scientific study conducted at the E.D. Goldberg Research Institute of Fipm of the Tomsk NIMC
The aim of the study is to develop a new drug with a highly effective effect aimed at combating Alzheimer's disease.
Materials and methods of research
The object of the study was the amount of polyprenols isolated from Siberian Fir (Abies sibirica Ledeb.), the Pine family (fam. Pinaceae).
The experiments were performed on 108 outbred sexually mature male stock mice with an average body weight of 28-30 g (at the age of 5 weeks) conventional, category 1, obtained from the Department of Experimental Biomodeling of the E.D. Goldberg Research Institute of Tomsk NIMC.
The design of the experiments and the maintenance of the animals were approved by the Ethics Committee of the E.D. Goldberg Research Institute of the Tomsk NIMC.
The antiamnestic activity of the substance - the sum of polyprenols was studied in the dose range 5, 20, 50, 100, 200, 500 mg/kg. An experimental model of Alzheimer's disease was created by intraperitoneal administration of scopolamine at a dose of 2 mg/kg [11] for 20 days. Choline alfoscerate (Gliatilin – Catalent Italy S.P.A.) was used as a comparison drug.
All the animals were divided into 9 groups of 12 individuals each. The first group (intact control) was injected intraperitoneally with saline throughout the experiment. The remaining groups were also injected intraperitoneally with scopolamine for 20 days, and from 21 to 30 days intraperitoneally with saline. In addition to saline, 0.4 ml of water was injected into the stomach of the second group (negative control). The third group received the comparison drug gliatilin at a dose of 90 g/kg from day 21 to 30 (positive control) [12]. Groups from the fourth to the ninth received the claimed remedy according to the indicated doses. On the 31st day of the experiment, 1 hour after the introduction of substances, a conditioned passive avoidance reflex (URPI) was developed in all animals. The reflex was checked 24 hours and 7 days after production.
The technique of the conditioned reflex of passive avoidance (URPI) is based on the suppression of the innate reflex of preference for dark space available in rodents [10]. The experimental setup consisted of a camera consisting of two compartments: a large one – illuminated and a small one - dark. The animal was placed in a light compartment and soon (after 10-20 seconds), due to the innate reflex of preference for dark space, it moved into a small compartment, after which the door connecting both compartments was blocked and an electric current was applied to the floor of the dark compartment, consisting of parallel alternating electrodes, with pulses of 50 Ms duration, frequency 5 Hz and amplitude 50 mA. After 10 seconds, the door was opened and the animal could jump out into a light compartment with an ordinary floor. As a result of the described procedure, the animals developed a conditioned reflex of avoiding dark space. When checking the reproducibility of the reflex, the animals were placed in a light compartment, in the corner opposite from the entrance to the dark compartment and observed for 3 minutes. The time of the first entry into the dark compartment (latent entry time) and the total time spent in the dark compartment were recorded.
Statistical processing of the obtained results was carried out using the software package "Statistica 6.0" for Windows [13].
Results and their discussion
In the course of the conducted studies, it was found that scopolamine administered once at a dose of 2 mg / kg for 20 days caused a distinct violation of the production and reproducibility of URPI. At the same time, the course administration of the substance - the sum of polyprenols for 5 days had an improving effect on the reproducibility of the reflex exceeding the effect of the comparison drug Gliatilin. A decrease in the effect of a substance in large doses, versus its effect in small doses, is a phenomenon quite common in the pharmacology of receptor drugs (see Table 1).
Table 1 – The effect of the course administration of the sum of polyprenols on the production of URPI according to the results of checking the reproducibility of the reflex 24 hours and 7 days after production in mice with impaired cognitive and amnestic processes caused by chronic (20 days) administration of scopolamine at a dose of 2 mg/kg (X±m; n=12)
Note: * - the probability of a null hypothesis is less than 0.05 compared to group 2 (scopolamine control)
The substance - the sum of polyprenols of Siberian Fir (Abies sibirica Ledeb.), the Pine family (fam. Pinaceae) has an antiamnestic effect on an experimental model of Alzheimer's disease caused by chronic (for 20 days) administration of scopolamine. The most pronounced maximum effect was observed at doses of 20 and 50 mg/kg.
Thus, according to the experimental data, the drug freely passes through the blood-brain barrier (BBB) and serves as a direct donor of the neurotransmitter acetylcholine in the presynaptic membranes of cholinergic neurons. When ingested by enzymes, it is broken down into choline and glycerophosphate. The resulting choline, due to its electrical neutrality, penetrates the blood-brain barrier and enters the brain, where it serves as the basis for the formation of acetylcholine. In addition, glycerophosphate, being a precursor of phospholipids of the neuronal membrane, stimulates the formation of phosphatidylcholine, which restores the phospholipid composition of neuronal membranes and improves their plasticity, positively affects the functional state of cell microstructures, improving the cytoskeleton of neurons, increasing the mass of mitochondria and ribosomes. The drug normalizes the transmission of nerve impulses, potentiates anabolic processes in neurons responsible for membrane phospholipid and glycerolipid synthesis, improving the state of cognitive and behavioral functions [1].
Conclusion
The above study shows that a new remedy with an antiamnestic effect has been found based on the sum of polyprenols of Siberian Fir (Abies sibirica Ledeb.), the Pine family (fam. Pinaceae). This drug can be used to treat diseases of neurodegenerative pathological processes, in particular Alzheimer's disease.
List of literary sources
1. Dzyak L.A., Tsurkalenko E.S. The role of cholinergic deficiency in the pathogenesis of neuropsychiatric diseases // International Neurological Journal. – 2019. – vol. 105, No. 3. – pp. 39-47.
2. Bartus R.T. The cholinergic hypothesis of geriatric memory disfunction / R.T. Bartus, R.L. Dean, B. Beer, A. Lippa. // Science. – 1982. – Vol. 217, № 4558. – P. 408–414.
3. Bullock R. New drugs for Alzheimer’s disease and other dementias // Br. J. Psychiatry. – 2002. – Vol. 180. – P. 135-139.
4. Galasko D. New approaches to diagnose and treat Alzheimer’s disease: a glimpse of the future // Clin. Geriatr. Med. – 2001. – Vol. 17, № 2. – P. 393 – 410.
5. Doggrell S.A. Treatment of dementia with neurotransmission modulation / S.A. Doggrell, S. Evans // Expert opin. Investig. Drugs. – 2003. – Vol. 12. – P. 1633 – 1654.
6. Bond M. The effectiveness and cost-effectiveness of donepezil, galantamine, rivastigmine and memantine for the treatment of Alzheimer's disease / M.Bond, G, Rogers, J. Peters, R Anderson, et al. (review of Technology Appraisal № 111): a systematic review and economic model // Health technology assessment (Winchester, England). – 2012. – Vol. 16, № 21. – P. 1–470.
7. Amenta F., Tayebati S.K., Vitali D., Di Tullio M.A. Association with the cholinergic precursor choline alphoscerate and the cholinesterase inhibitor rivastigmine: an approach for enhancing cholinergic neurotransmission // Mech. Ageing. Dev. – 2006. –Vol. 127. –P. 173–179
8. Sato N., Sakamori M., Haga K., Takehara S., Setoguchi M. Antagonistic activity of Y-25130 on 5-HT3 receptors // Jpn. J. Pharmacol. – 1992. –Vol. 59,№ 4. – 443-448.
9. Shabanov P. D., Borodkin Yu. S. Memory disorders and their correction. – L.: Nauka. – 1989. – 127 c.
10. Buresh Ya. Methods and basic experiments for studying the brain and behavior. (translated from English. edited by prof. A. S. Batuev) / Ya Buresh, O. Bureshova, J. P. Houston). M.: Higher School, 1991. -398 P.
11. Guidelines for conducting preclinical studies of medicines. Part one. — Moscow : Grif and K, 2012. – p. 291.
12. Hyun Mi Kim Effects of Choline Alfoscerate and Memantine on Memory Improvement of Scopolamine-induced Memory Impairment animal Model of Alzheimer’s Disease / Hyun Mi Kim, SeongHee Kang, Seong-Wan Cho, Bo Sun Kang // YakhakHoeji. – 2017. – Vol. 61 № 6. – P. 292-300.
13. Lakin G. F. Biometrics: Textbook for biol. spec. universities. Moscow: Higher School, 1990. – p. 352.
The substance - the sum of polyprenols of Siberian Fir (Abies sibirica Ledeb.), the Pine family (fam. Pinaceae) has an antiamnestic effect on an experimental model of Alzheimer's disease caused by chronic (for 20 days) administration of scopolamine. The most pronounced maximum effect was observed at doses of 20 and 50 mg/kg.
Thus, according to the experimental data, the drug freely passes through the blood-brain barrier (BBB) and serves as a direct donor of the neurotransmitter acetylcholine in the presynaptic membranes of cholinergic neurons. When ingested by enzymes, it is broken down into choline and glycerophosphate. The resulting choline, due to its electrical neutrality, penetrates the blood-brain barrier and enters the brain, where it serves as the basis for the formation of acetylcholine. In addition, glycerophosphate, being a precursor of phospholipids of the neuronal membrane, stimulates the formation of phosphatidylcholine, which restores the phospholipid composition of neuronal membranes and improves their plasticity, positively affects the functional state of cell microstructures, improving the cytoskeleton of neurons, increasing the mass of mitochondria and ribosomes. The drug normalizes the transmission of nerve impulses, potentiates anabolic processes in neurons responsible for membrane phospholipid and glycerolipid synthesis, improving the state of cognitive and behavioral functions [1].
Conclusion
The above study shows that a new remedy with an antiamnestic effect has been found based on the sum of polyprenols of Siberian Fir (Abies sibirica Ledeb.), the Pine family (fam. Pinaceae). This drug can be used to treat diseases of neurodegenerative pathological processes, in particular Alzheimer's disease.
List of literary sources
1. Dzyak L.A., Tsurkalenko E.S. The role of cholinergic deficiency in the pathogenesis of neuropsychiatric diseases // International Neurological Journal. – 2019. – vol. 105, No. 3. – pp. 39-47.
2. Bartus R.T. The cholinergic hypothesis of geriatric memory disfunction / R.T. Bartus, R.L. Dean, B. Beer, A. Lippa. // Science. – 1982. – Vol. 217, № 4558. – P. 408–414.
3. Bullock R. New drugs for Alzheimer’s disease and other dementias // Br. J. Psychiatry. – 2002. – Vol. 180. – P. 135-139.
4. Galasko D. New approaches to diagnose and treat Alzheimer’s disease: a glimpse of the future // Clin. Geriatr. Med. – 2001. – Vol. 17, № 2. – P. 393 – 410.
5. Doggrell S.A. Treatment of dementia with neurotransmission modulation / S.A. Doggrell, S. Evans // Expert opin. Investig. Drugs. – 2003. – Vol. 12. – P. 1633 – 1654.
6. Bond M. The effectiveness and cost-effectiveness of donepezil, galantamine, rivastigmine and memantine for the treatment of Alzheimer's disease / M.Bond, G, Rogers, J. Peters, R Anderson, et al. (review of Technology Appraisal № 111): a systematic review and economic model // Health technology assessment (Winchester, England). – 2012. – Vol. 16, № 21. – P. 1–470.
7. Amenta F., Tayebati S.K., Vitali D., Di Tullio M.A. Association with the cholinergic precursor choline alphoscerate and the cholinesterase inhibitor rivastigmine: an approach for enhancing cholinergic neurotransmission // Mech. Ageing. Dev. – 2006. –Vol. 127. –P. 173–179
8. Sato N., Sakamori M., Haga K., Takehara S., Setoguchi M. Antagonistic activity of Y-25130 on 5-HT3 receptors // Jpn. J. Pharmacol. – 1992. –Vol. 59,№ 4. – 443-448.
9. Shabanov P. D., Borodkin Yu. S. Memory disorders and their correction. – L.: Nauka. – 1989. – 127 c.
10. Buresh Ya. Methods and basic experiments for studying the brain and behavior. (translated from English. edited by prof. A. S. Batuev) / Ya Buresh, O. Bureshova, J. P. Houston). M.: Higher School, 1991. -398 P.
11. Guidelines for conducting preclinical studies of medicines. Part one. — Moscow : Grif and K, 2012. – p. 291.
12. Hyun Mi Kim Effects of Choline Alfoscerate and Memantine on Memory Improvement of Scopolamine-induced Memory Impairment animal Model of Alzheimer’s Disease / Hyun Mi Kim, SeongHee Kang, Seong-Wan Cho, Bo Sun Kang // YakhakHoeji. – 2017. – Vol. 61 № 6. – P. 292-300.
13. Lakin G. F. Biometrics: Textbook for biol. spec. universities. Moscow: Higher School, 1990. – p. 352.
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