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DOI 10.26724/2079-8334-2018-4-66-172-175 UDC 615.01.547:615.461.2
INFLUENCE OF OXAMINE ACID DERIVATIVES ON THE SECRETORY
FUNCTION OF KIDNEYS
E-mail: [email protected]
The purpose of this project was to experimentally study the effect of new dicarboxylic acids derivatives on the excretory function of the kidneys. The studied compounds under the water loading conditions have a multidirectional effect on the kidneys excretory function in white rats of the Wistar line. The diuretic activity of the studied compounds was influenced both by the chemical structure of the substances and by its location. As a result of the performed research, substances increasing diuresis with the diuretic activity exceeding that of the reference preparation - hypothiazide - were found. Among the studied compounds, there were substances with an antidiuretic effect. Replaced arensulfoniloxamine acid amides are the perspective group of compounds for further pharmacological study with the aim of development of new medicines with diuretic activity on their basis.
Key words: dicarboxylic acids derivatives, oxamine acid derivatives, excretory function of the kidneys, diuretic activity, antidiuretic effect.
The work was carried out within the framework of the research program of the National Pharmaceutical University on the problem of "Creation of New Drugs" (State Registration Number 0198U007008).
An urgent problem of modern pharmacology is the creation of new, more effective and less toxic Ukrainian drugs for treating diseases of the kidneys and urinary tract [2].
Pharmacological correction of kidneys activity is performed in many diseases of the kidneys and urinary tract, cardiovascular failure, some forms of liver pathology, etc. [6].
There are many groups of diuretic drugs with different acting mechanisms used in practical health care today. The mechanisms of kidney function regulation, intrarenal hemodynamics, inter- and intra-pharyngeal relationships, electrochemical ion transport processes are still being studied [5].
The study of mechanisms regulating kidneys function is the basis for searching new substances to create new highly effective drugs that regulate the function of the kidneys [9].
Perennial synthetic and biological studies at the National Pharmaceutical University have allowed the accumulation of large material on the structural and pharmacological properties of dicarboxylic acid derivatives. The latter comprise substances with anti-inflammatory, antimicrobial, sedative, hypoglycemic, diuretic and other effects [4, 7]. This was the prerequisite for studying diuretic action of new compounds of the sulfamide series synthesized at the National Pharmaceutical University.
The purpose of this study was to experimentally study the effect of new dicarboxylic acids derivatives on the excretory function of the kidneys.
Materials and methods. To solve this problem, 40 new chemical substances were used as the object of research. They were substituted amides of arenesulfonyloxamic acids, first synthesized at the Pharmaceutical Chemistry Department of the National Pharmaceutical University.
The structure of these compounds was confirmed using modern physicochemical methods of elemental analysis, UV, IR, PMR, and mass spectrometry, counter-synthesis, and the purity of the synthesized substances was monitored by thin-layer chromatography.
The studied compounds are white crystalline substances of basic character, odorless, with a clear melting point, soluble in polar organic solvents, solutions of caustic bases, mineral acids. These synthetic derivatives were administered to laboratory animals in the form of aqueous solutions or 3-5% finely dispersed aqueous suspension stabilized by Tween-80, which is a hydroxyethylation product of monooleate sorbitan (VFS-42-167-72). All manipulations with laboratory animals were carried out in accordance with the provision on the use of animals in biomedical research (Strasburg, 2005) and "The general ethical principles of experiments on animals", approved by the Fifth National Congress on Bioethics (Kyiv, 2013).
The effect of these substances on the kidneys excretory function was studied on white Wistar male rats weighing 160-190 g by the method of Berkhin [1,8]. To study the diuretic action, we used series of animals, 7 rats in each group. To study aqueous diuresis, rats were kept on a constant diet with free access to water. Before giving water, the animals were kept for 2 hours without food and water. Then the rats were injected the studied substances with a probe into the stomach. These substances were at a dose of 0.0050.01 LD50 in the form of an aqueous suspension simultaneously with a water loading of 3 ml per 100 g of body weight of the animal. Urine was collected every hour for 4 hours. Comparison drugs were widely
© O.N. Litvinova, V.S. Litvinov, 2018
used in clinical practice, standard drugs Furosemide at a dose of 20 mg/kg, Hypothiazid in a dose of 50 mg / kg and Adiurecrin in a dose of 10 mg/kg.
Table
The effect of substituted amides of arenesulfonyloxamic acids on the kidneys excretory
function in white Wistar rats
Compound Number Dose, mg/kg Diuresis
for 2 hours by the control time, in % for 4 hours by the control time, in %
/ M±m / ,ml / M±m / ,ml
1 2 3 4 5 6
1 19.5 2.34±0.22* 243.7 4.98±0.09* 198.4
2 15.4 0.52±0.09* 61.5 1.37±0.09* 54.6
3 17.2 0.78±0.13* 81.2 1.45±0.06* 57.8
4 22.8 1.28 ± 0.12 133.3 2.57±0.06 102.4
Control - 0.96 ± 0.05 100 2.51±0.06 100
5 16.4 1.78±0.11* 148.3 3.44±0.11* 150.2
6 17.0 1.29 ± 0.18 107.5 2.96 ± 0.06 129.2
7 13.4 1.14 ± 0.21 95.0 2.67 ± 0.08 116. 9
8 14.2 1.44 ± 0.17 120.0 2.54 ± 0.28 110.8
9 13.8 0.77±0.11 * 64.2 1.83±0.18 * 79. 9
10 17.2 0.84±0.09 * 70.0 1.92 ± 0.07 83.8
11 21.0 1.21 ± 0.11 100.8 2.44 ± 0.06 106.5
12 22,6 1.44 ± 0.18 120.0 2.36 ± 0.09 103.1
13 20.6 2.37±0.22 * 197.5 4.14±0.11 * 180.8
14 21.4 1.86 ± 0.17 155.0 3.42±0.12 * 149.3
Control - 1.20 ± 0.11 100 2.29±0.14 100
15 45.5 0.91 ± 0.11 96,8 2.11 ± 0.16 81.7
16 15.0 1.04 ± 0.16 110.7 2.96 ± 0.11 114.7
17 25.5 2.04±0.07 * 217.0 4.28±0.17 * 165. 9
18 42.3 2.55±0.09 * 271.3 4.92±0.22 * 190.7
19 22,8 1.59±0.09 * 169.1 2.67 ± 0.18 103.5
20 i, 8 0.84±0.06 89.4 1.92±0.16 * 74.4
21 30.5 1.70±0.08 * 180,8 3.10 ± 0.11 120.2
22 29.3 1.63±0.06 * 173.8 4.36±0.08 * 168.8
23 26.3 1.16±0.08 123.8 3.68±0.11 142.8
Control — 0.94±0.04 100 2.58±0.11 100
24 23.0 1.54 ± 0.13 126.2 2.84 ± 0.17 122. 9
25 53.0 2.14 ± 0.14 175.4 5.44±0.23 * 235.5
25 20.5 1.68 ± 0.12 137.2 2.74 ± 0.22 118.6
27 45.3 1.32 ± 0.07 108.1 2.64 ± 0.13 114.3
28 61.2 1.12 ± 0.13 91.8 2.69 ± 0.13 116.4
29 53.4 1.48 ± 0.17 121.3 3.35 ± 0.19 145.0
30 50.4 1.28±0.08 104 9 2.24±0.11 96 9
31 51.0 0.84±0.06 * 68.8 1.64±0.07 * 71.0
Control - 1.22 ± 0.11 100 2.31±0.09 100
32 41.5 1.53 ± 0.12 130.7 3.14 ± 0.18 123,6
33 40.8 0.88±0.09 * 75.2 1.93±0.13 * 75.9
34 45.0 1.41 ± 0.18 120.5 2.88 ± 0.16 113.4
35 52.0 0.81±0.14 * 69.2 1.69±0.17 * 66.5
36 48.4 1.61 ± 0.13 137.6 2.82 ± 0.13 111.0
37 42.4 1.34 ± 0.18 114.5 3.48 ± 0.07 137.0
Control - 1.17±0.06 100 2.54±0.13 100
38 40.8 1.33 ± 0.12 123.1 3.05 ± 0.19 128.1
39 32.8 1.24 ± 0.09 114.8 3.04 ± 0.16 127.7
40 25.7 2.90±0.27 * 268.5 4.91±0.17 * 206.3
Control - 1.08±0.07 100 2.38±0.11 100
Hypothiazid 50.0 2.01± 0.14* 164.7 4.20±0.21* 164.1
Furosemide 20.0 3.8±0.19* 311.4 8.40±0.27* 328.1
Adiurecrin 10.0 0.7±0.12* 57.3 1.10±0.14* 42.9
Control - 1.22±0.13 100 2.56±0.17 100
Note: the "*" sign indicates the reliability of the differences with the control, p <0.05
The data of experimental studies were processed by common methods of variation statistics using Student's t-test. The difference was considered statistically significant at a confidence level of p <0.05 [ 3].
Results of the study and their discussion. The obtained results analysis shows that most of the studied substances under conditions of water stress cause an increase in the kidneys excretory function (table).
Thus, among the N-acyl-N-arylaminoethylamidoranesulfonyloxamic acids (Compounds 1-15), the most active was Compound 1, which, in a dose of 19.5 mg/kg, increased diuresis by 143.7% for 2 hours, and for 4 hours - by 98.4%. In its structure, this substance has a p-amine and phenyl (Compound 1) radicals, and their replacement by an n-methyl (Compound 4) radical leads to a decrease in diuretic properties. Thus, Compound 4 increases diuresis for 2 hours by 33.3%, and for 4 hours - by only 2.4%. Compounds 11 and 12 were practically inactive. They combine methyl, phenyl radicals and a hydrogen atom (Compound 11), as well as 2-methyl and phenyl (Compound 12) radicals in their structure. The introduction of the chloromethyl radical (Compounds 2 and 3) into the structure of N-acyl-N-arylaminoethyl-amidoranesulfonyloxamic acids led to antidiuretic activity. These substances reduced diuresis in rats for 4 hours by 45.4 and 42.2%, respectively.
The majority of N-R-substituted amides of arenesulfonyloxamic acids (Compounds 16-28) caused an increase in urinary excretion by 14.3-135.5%. The most active were Compounds 18 and 25, combining 4-aminophenyl and g-hydroxypropyl (Compound 18), 4-carbomethoxyaminophenyl and 5-carboxyamyl (Compound 25) radicals in their structure. These compounds increase diuresis for 4 hours by 90.7 and 135.5%, respectively. The introduction of benzyl (Compound 17), heptyl (Compound 20) and carboxymethyl (Compound 21) radicals into the oxamide part of the molecule causes a decrease in diuretic activity. The antidiuretic effect was observed in Compounds 16 and 20, combining 4-aminophenyl and ethyl (Compound 16), 4-aminophenyl and heptyl (Compound 20) in their structure, they reduced urination for 4 hours by 18.3 and 25, 6%, respectively.
Among the N-substituted amides of 4- (R-benzamido) -benzenesulfonyloxamic acids (Compounds 29-40), most substances increased diuresis by 11-106.3%. The most active was Compound 40 containing a nitro group in the benzene ring, and a hydroxyl radical in the side chain. This compound, in a dose of 25.7 mg/kg, increases diuresis by 168.5% for 2 hours, and by 106.3% for 4 hours. Compounds 30, 31, 33 and 35 caused a decrease in diuresis by 3.1-33.5% on average.
Thus, the performed studies of the effect of substituted amides of arenesulfonyl-amino acids in conditions of water loading showed that most of the studied compounds caused an increase in the kidneys excretory function in the rats taking part in the experiment. Among the studied substances, the most pronounced diuretic effects occurred in the Compounds 18 and 40, which in doses of 42.3 mg/kg and 25.7 mg/kg increase the diuresis for 2 hours by 171.3 and 168.5%, respectively. We also found substances (Compounds 2 and 3) that exhibit pronounced antidiuretic activity. These compounds reduce diuresis in rats for 4 hours by 45.4 and 42.2%, respectively.
1. Substituted amides of arenesulfonyloxamic acids under the conditions of water loading have a multidirectional effect on the kidneys excretory function in white rats of the Wistar line. The diuretic activity of the studied compounds was influenced both by the chemical structure of the substituent and by its location.
2. Among all studied substances, the most pronounced diuretic effect was provided by Compounds 18 and 40. Their diuretic effect was significantly superior to the Hypothiazide, but inferior to Furosemide.
3. Among the studied compounds, there were substances with an antidiuretic effect. Compounds 2 and 3 are equal to Adiurecrin in their antidiuretic effect.
Prospects for further research: sufficient diuretic activity of most substituted amides of arenesulfonyloxamic acids makes them promising substances for further targeted synthesis and pharmacological screening with the goal of creating drugs with diuretic properties on their basis.
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ВПЛИВ ПОХ1ДНИХ ОКСАМ1НОВИХ КИСЛОТ НА ЕКСКРЕТОРНУ ФУНКЦ1Ю НИРОК
Литвинова О. М., Литвинов В. С.
Метою роботи було вивчення впливу нових похщних дикарбонових кислот на видшьну функщю нирок у бших щурiв в експеримени. Вивченi сполуки мають рiзноспрямовану дш на видiльну функцiю нирок у тварин. На дiуретичну активнiсть вивчаемих сполук впливала як хiмiчна будова замюника, так i його розташування. Виявленi речовини, якi викликають збшьшення дiурезу, що перевищуе по дiуретичнiй активностi еталонний препарат порiвняння - ппоиазид. Знайденi сполуки, якi виявляли достатню антидиуретичну активнiсть. Замiщенi амщи аренсульфонiлоксаминових кислот е перспективною групою сполучень для подальшого фармакологiчного вивчення з метою створення на !х основi лiкарських препаратiв з дiуретичними властивостями.
Ключовi слова: похiднi дикарбонових кислот, похщш оксамiнових кислот, екскреторна функщя нирок, дiуретична дiя, антидiуретична актившсть.
Стаття надiйшла 28.02.18 р.
ИЗУЧЕНИЕ ВЛИЯНИЯ ПРОИЗВОДНЫХ ОКСАМИНОВЫХ КИСЛОТ НА ЭКСКРЕТОРНУЮ ФУНКЦИЮ ПОЧЕК Литвинова О. Н., Литвинов В. С.
Целью работы было изучение влияния новых производных дикарбоновых кислот на выделительную функцию почек в эксперименте у белых крыс. Изученные вещества оказывают разнонаправленное действие на выделительную функцию почек у животных. На диуретическую активность изучаемых соединений влияла как химическая структура заместителя, так и его расположение. Выявлены вещества, вызывающие увеличение диуреза и превышающие по диуретической активности эталонный препарат сравнения - гипотиазид. Обнаружены также соединения, проявляющие достаточную антидиуретическую активность. Замещенные амиды аренсульфонилоксаминовых кислот являются перспективной группой соединений для дальнейшего фармакологического изучения с целью создания на их основе лекарственных препаратов с диуретическими свойствами.
Ключевые слова: производные дикарбоновых кислот, производные оксаминовых кислот, экскреторная функция почек, диуретическое действие, антидиуретическая активность.
Рецензент: Костенко В.О.
DOI 10.26724/2079-8334-2018-4-66-175-180 УДК: 611.451-018:547.96]-019-013:616.441-008.6
1МУНОГ1СТОХ1М1ЧНЕ ДОСЛ1ДЖЕННЯ НАДНИРКОВИХ ЗАЛОЗ ПОТОМСТВА ЩУР1В, ЩО РОЗВИВАЛОСЯ ЗА УМОВ ЕКСПЕРИМЕНТАЛЬНОГО Г1ПО- ТА Г1ПЕРТИРОЗУ
МАТЕРИНСЬКОГО ОРГАН1ЗМУ
E-mail: [email protected]
З використанням iмуногiстохiмiчних методiв - визначення MapKepiB клтнно! прoлiферацii та апоптозу Ki-67, VEGF та Casp3 вiдповiдно - дослщжено вплив експериментального rino- та ппертирозу материнського органiзму на надниркoвi залози потомства щурiв 1-1 та 10-1 доби постнатального розвитку. Продемонстровано високу штенсившсть прoцесiв як прoлiферацii, так i апоптозу, що супроводжували постнатальний морфогенез надниркових залоз; при цьому найбшьш активна перебудова iдентифiкoвана у пучковш зoнi кори. На 1-у добу постнатального онтогенезу на rai гтотирозу виявлено затримку розвитку мозково! речовини, тoдi як гiпертирoз супроводжувався пригшченням прoлiферативнol активнoстi клiтин клубочково! зони наднирникiв. На 10-у постнатальну добу на rai тиро!дного дисбалансу задокументовано посилення процеЫв прoлiферацil i апоптозу у складi юрково1 речовини у поеднанш з пригнiченням штенсивносп обох прoцесiв у мoзкoвiй речовиш надниркових залоз.
Ключовi слова: щури, онтогенез, надниркoвi залози, материнський ппо- та гiпертирoз, iмунoгiстoхiмiчне дослщження.
Робота е фрагментом НДР «Лектино- та iмуногiстохiмiчний aHan.i3 вуглеводних детермшант нормальних та патологiчно змiнених клтин i тканин» (№ державноi реестрацн 0117U001076).
Порушення функци щитоиод16ио1 залози належать до найпоширешших захворювань, охоплюючи близько 3% населения св1ту [1]. Численш сиостережеиия свщчать про вагомий вплив тиро1дних гормошв иа розвиток i фуикцюиуваиия надниркових залоз - як безпосередньо, так i через гшоталамо-гшоф1зарио-адреиалову вюь [12]. Гормоии щитоиод16ио1 залози i иадиирииюв вщграють ключову роль у забезиечеиш виутршиьоматкового гомеостазу, диференщацп i дозр1ваиия оргашв плода у вщповщносп до часу гестаци [10, 3].
У иоиередшх дослщженнях [4-8] з використанням метод1в класично1 пстологи, морфометрп та лектииово1 пстох1ми 6уло показано, що гшотироз материнського оргашзму шдукуе затримку розвитку надниркових залоз потомства, тод1 як ппертироз ирискорюе 1хиш розвиток, обумовлюючи гшертрофда юрково1 речовини. Аиал1з достуиио1 л1тератури виявив вщсутшсть иублшацш, як 6 характеризували змши ирол1феративио1 активиост i явищ аиоитозу у надниркових залозах потомства, що розвивалося за умов дисбалансу тиро1дних гормошв материнського оргашзму.
© С.О. Луцик, А.М. Ященко, 2018
