UA139049U - N-BENZYLAMINOMETHANSULPHOIC ACID AS AN ANTI-INFLUENZA AGENT - Google Patents

Abstract

N-Benzylaminomethanesulfonic acid of the formula: as an anti-influenza agent.

Description

M-Benzylaminomethanesulfonic acid (VpAM5A) of the formula: / (c) ac / on (c) that exhibits a certain level of antiviral activity and can be used in the pharmaceutical industry for the development of anti-influenza drugs.

Influenza is the most widespread infection among those representing a significant socio-economic problem on an international scale. Its most important characteristics are its epidemic nature and high mortality, especially in pandemic conditions. Despite the successes achieved in the field of chemotherapy and vaccine prevention of influenza, it remains a difficult-to-control infection due to high genetic variability and complications of various etiologies after the acute stage of the disease, which often lead to a fatal outcome. New strains of influenza viruses that constantly appear and circulate in the human population are dangerous human pathogens. Today, the protocol for the treatment of influenza, recommended by the WHO, contains drugs of two groups: the first - derivatives of the adamantane series (amantadine, rimantadine), which are blockers of M2 channels and are effective only against influenza A viruses. The second group of drugs effective against influenza A viruses and B, include neuraminidazk inhibitors (zanamivir, oseltamivir) (see M/NO (2010 Rergiagu) MUNO aScideiipev og RNaptasoiodisai!

Mapadetepi oi Rapadetis (NIMI) 2009 Ipytsepla ap oiShek IpbPepla Migpize5. OVI: pOr/Lymly lupo,Ipu/svg/gezoigsev/rybriyasatopv/vmipeyish/pIp!I ive apii-migaizye20090820/ep/Ipdeh. sweat

Unfortunately, the vast majority of human isolates of influenza viruses quickly acquire resistance to the action of both groups of anti-influenza drugs - rimantadine, oseltamivir (see Vgidni V.A., ZNau 0.,

Zpy V., Soh M.U., Kiitom A.I. Adatapiape geziziapse atopa ipishepla A mogizez isoiiaied vapu digipd

She 2005-2006 ipishepla zeazop ip She Opaieyd biaiez 3. At. Honey. Av5os 2006. Mine. 295, M 8. R. 891-894. NOrz//doi.oga/10.1001/ata.295.8.00660020; Suragemha to them. Naudep B... M2 apa peshatipidaze ippirpogv: api-ipPYashepga asiimpu, tesNapivit5 oi gevibyapse apa siipisa! epesiimepev5.

Ip: Kauzhshaoka U, eayog. IpiYAShepla migoiodu: sitepi (orish. Mutopanat, Opiped Kipddot: Saivieg

Rge55; 2006. R. 169-202.

In 2013, results were published that indicated that the use of neuraminidase inhibitors for the treatment of a patient infected with a drug-resistant H7MO influenza strain contributed to the reproduction and spread of the resistant strain in particular (Mep N.-I.., MeKitt-Vgezsnkip u).

Ko) H., Spou K.-T., Mopd 0. 0. M., Snetspa R. R. N., 7poi U., Ma I. N., 7py N., MUerbu V. u., Scap U .,

M/erzieg V. S., Reigiza 9. 5. M. A Veviviapse o Meshgatipidaze Ippirioge Sopieited Bu ap H2g92K

Mshaiiop ip a Nytap Ipshepga Migy5 H7MO Izoiaie Sap Ve MazkKea bu a Mikhea V/K Miga! Rorshiyop. tvio, 2013; 4 (4): - e00396-13. - Rybriyivney oopipe 2013 shu 16 Asseb55 tod: per//tbryo.azt.Oga/sopiepi/4/4/е00396-13.5НопП1.

Aminoalkanesulfonic acids are an important class of M-,5-containing organic compounds, the interest in which is due to their specific physicochemical properties (in particular, the pKa values of these acids are in the physiological pH range) (see Khoma R.E., Osadchii L .T., Dlubovsky R.M. Aminomethanesulfonic acid and ee

M-products - components of buffer solutions N. Huda. ONU Bulletin. Chemistry. 2015. Vol. 20, Mo. 3.

pp. 66-75) and a wide spectrum of biological activity. It is known that M-derivatives (aminomethanesulfonic acid) AM5BA have antitumor and antiviral effects (see

MeeiaKapiap I.O., Napypud M.N. a-AtipoaiKapezyMopis Asidb. U. Ogu. Spent 1959. MoI24, M 12.R. 1943-1948).

M-derivatives of AM5A, their salts are biologically active compounds that exhibit antimiotic, cytostatic, bactericidal properties and can be used as bactericidal and insecticidal preparations (see Stuagpom R.6I., Kaiaema O.M., Mashtoma O.E., Myvip V.7., AIchopbom

MA. VAeasiop o oi 0 8- itipoaisoyoiv om/yN o by o aiokhide o Bupiyebviv oi 00S(t)-(2-

NuagohuaiKuiatipo)rpnepu(izorgorui)-teiapesiopis asidv5. Ex. 5 U. Sep. Spent 2010. Mine. 80, M 4.

R. 761-764). Some o-aminoalkanesulfonic acids show antistaphylococcal and

BO antifibrinolytic activity (see Ecii A. SbookK E.5. Rgobiois5. Apiiztarpuiosossa! apa apiyirhipoiuis asiimtsievz oi o-atipo-apa o-dpapidipoaiKapezy pis asidv. /). Medicine. Snet 1975. Mine. 18, M 5. P 502-505. BO1:10.1021//t00239a013; Patent of Ukraine for utility model OA 127587.

IPC С07С 309/00 Aminomethanesulfonic acid and its M-alkylated derivatives as antistaphylococcal agents. / Ennan A.A., Khoma R.E., Hrydina T.L., Fedchuk A.S. Mo in 201802703; statement 16.03.2018; published 10.08.2018, Bul. Mo 15.).

For AM5A (I) and its M-methyl- (II), M-(2-hydroxy)ethyl- (PP), M-tert-butyl (IM) and M-benzyl (M) derivatives (in the form of sodium salts) previously, acute toxicity was assessed in male rats after oral and parenteral administration (see Khoma R.E., Znnan A.A., Hrydyna

T.L., Fedchuk A.S, Lozytskyi V.P., Godovan V.V., Antonenko P.B. Synthesis, structure, physicochemical characteristics and biological activity of aminomethanesulfonic acids. Materials

MI of the National Congress of Pharmacists "Pharmacy of the 21st century: trends and prospects". Kharkiv, September 13-16, 2016. Vol. 1. P. 54-55 According to the results of the study of the compound salt !! when administered intraperitoneally in a dose of 2000 mg/kg, it is classified as practically non-toxic substances. Oral administration of Ma salts I - M in a dose of 5000 mg/kg did not cause the death of animals, that is, the studied compounds with this route of administration are classified as practically non-toxic substances.

Ethyl (ZE, 48, 55)-5-amino-4-acetamido-3-(pentan-3-yloxy)cyclohex-1-ene-Icarboxylate (Oseltamivir , Tamiflu) (see the State Register of Medicinal Products of Ukraine.

OVS pirulumlu.a.sot.tsa.). The disadvantages of this compound are the rather high cost, multi-stage synthesis, possible side effects in the children's age group and use in the age group after 5 years.

This compound was selected as the closest analogue.

The choice of the analogue is due to common features with the proposed useful model - common structural fragments (the presence of an amine fragment), biological action (anti-influenza activity).

The proposed useful model is based on the task of expanding the range of compounds that have a certain level of anti-influenza activity.

The task was solved by using VpAM5BA, obtained by us according to the original method (see Patent of Ukraine for a utility model OA 75340, IPC СО7С 309/00, 309/15 Method of obtaining M-derivatives of aminomethanesulfonic acid. Khoma R.E., Ennan A.A .-AND.,

Koroeva L.V., Lavreka O.O., Helmboldt V.O. Me and 201206510; statement 05/29/2012; published 26.11.2012, Bull. Mo 22) (reactions 1, 2) as an anti-influenza agent.

Кк зЗПАМНОЗ(СНоО я MЯзтно no t ТЙ В - 5 5 5 A

Kk

She

Kk and Z "ZOZ -k.:3 va n | x ra aa o) on

Kk Kk

Zo U (2) where I is SeH5СН» (M).

What is new in the useful model is that the obtained compound (VpPAM5A) of the general formula (c) p

FI; And it (c) showed a certain level of anti-influenza activity, comparable to the reference drug (see table).

The causal relationship between the structure of the object, the anti-influenza properties of which are claimed, and the obtained technical result is as follows: the use of practically non-toxic, relatively cheap and easily synthesized (in two stages) VPAM5A allows obtaining a certain level of anti-influenza activity, comparable to the reference - drug oseltamivir, which is currently a pharmaceutical anti-influenza drug. Synthesis

VpAM5A was carried out according to the original method (see Patent of Ukraine for utility model SHA 75340, МПК СО07С 309/00, 309/15 Method of obtaining M-derivatives of aminomethanesulfonic acid. Khoma R.E., Ennan A.A.-A., Koroeva L .V., Lavreka O0.0O., Helmboldt V.O. Mo and 201206510; application 05/29/2012; official gazette 11/26/2012, Bull. Mo 22).

Example 1.

The effect of drugs on the reproduction of influenza viruses A/Hong Kong/1/68 (NZM2) was studied using tissue culture of chorionallantoic membranes (HAOS) of 11-12-day-old chicken embryos. Methodological recommendations:

Ordinance A. V. Stefanov. - K.: Avicenna, 2002. - P.395-420; Maltseva A.I. Reproduction of influenza viruses in the culture of HAO tissue attached to the shell / A.I. Maltseva, V.E. Agranovskaya,

Ya.S. Shvartsman // Lab. business - 1973. - Mo11. - P.689-690), since this culture can be considered the closest to the level of a whole organism, which is a chicken embryo.

The effect of drugs on the reproduction of influenza viruses was studied by first dissolving the chemical compounds in DMSO (the final concentration was 1 95), and then in a glucose-gelatin support medium. Before conducting the experiments, the pH of the solutions of the studied drugs was checked and this indicator was adjusted to the physiological value of pH 7.2-7.4. After adjusting the pH of the solutions to a physiological value, the anti-influenza activity of the drugs was determined.

The effect of drugs on the reproduction of influenza viruses was studied by preparing a dilution of the virus-containing liquid on a glucose-gelatin medium that contained the drug under study (experiment) or did not contain this drug (control). The obtained dilutions were used to infect the fragments of CHAOS attached to the shell and placed in the holes of the polystyrene panels.

After thermostating for 24 hours at 37 "С, the titer of the infectious virus was determined in the control and experimental samples.

Titration of the influenza virus after exposure to the virus of the drug under study was carried out on the tissue culture of HAO. Tenfold dilutions of the samples were used to infect HAO fragments placed in the wells of polystyrene panels. After 48 hours of thermostating at 37", the presence of the virus in the wells was determined with the results of the hemagglutination reaction (HA).

The calculation of TID5o in experiments was carried out according to Kerber's modified method

Ashmarin | Ashmarin I.P. Calculates EdDvo with a small number of domestic animals /

From I.P. Ashmarin // J. microbiol. - 1959. - Mo 2. - P. 102-108) according to the formula: 19TIDvo--Y -4(5-0.5), where I is the initial dilution in the experiment; a- the difference between consecutive 19 dilutions; 5- the sum of the proportions of test objects that gave a positive result.

The statistical significance of antiviral action in tissue culture was determined using the non-parametric sign criterion (E.V. Gubler, A.A. Genkin. Application of non-parametric statistics criteria in medical and biological research. - L., 1973.

The obtained results (Table 1) indicate that VpAM5A at a final concentration of 10 mmol/l (which is 201.25 μg/ml) statistically significantly inhibits the reproduction of human influenza virus A/Hong Kong/1/68 (НЗМ2) in culture CHAOS tissues by 4.08 TaTiIDv»o compared to the control.

The reference drug (oseltamivir) at a final concentration of 10 mmol/l (which is 410 μg/ml) inhibited virus reproduction by 4.07 1aTIDvo. That is, anti-influenza activity

VpAM5A on KAOS tissue culture is similar to the action of the reference drug.

Example 2.

The effect of VpAM5BA and the reference drug on the reproduction of influenza virus A/RA/8/34 (NIMI) was determined in the culture of cells of the Khaos tissue similarly to Example 1.

In relation to influenza virus A/RA/8/34 (NIMI), the activity of VpAM5A was lower. This drug suppresses reproduction by only 1.67 TOTID»o compared to the control. While the reference drug inhibited the reproduction of this virus strain by 4.07 1aTIDvo compared to the control. That is, the effect of the studied drug was two times lower than the reference drug.

Example 3.

The effect of the drugs on the extracellular influenza virus A/RH/8/34 (NIMI) was determined as follows. The virus-containing liquid was diluted to 107 on a glucose-gelatin medium that contained (test) or did not contain (control) the drug. After that, the samples were kept at 4 "C for 20 hours, then for 2 hours at 37 "C, and the infectious titer was determined in them by infecting with tenfold dilutions the fragments of HAO tissue attached to the shell and located in polystyrene panels. After 48 hours of thermostating at 37", the titer of the virus was determined in RHA.

A.M. Reproduction of influenza viruses in the culture of HAO tissue attached to the shell /

A.I. Maltseva, V.E. Agranovskaya, Y.S. Shvartsman // Lab. business - 1973. - Mo 11. - P.689-690)1.

The obtained results indicate that in this experimental model, VpAM5A showed a slight effect on the extracellular influenza virus in the studied concentrations, in contrast to the reference drug, which did not exhibit virulicidal properties at all, without affecting the extracellular virus.

Table

Antiviral activity of VPAM5A against influenza viruses A/Hong Kong/1/68 (NZM2) and

A/RNA/V/ZA (NIMI) on tissue culture of CHAOS in comparison with the reference drug (oseltamivir)

Molar mass, ATatiID»o

MTV, minor

VpAM5A 201.25 4.08--0.5 1.6750.43 0.6720.3 4.07-0.562 | 40740.5 | -0.1740.3 "АТатИДё(1) - average indicator of inhibition of viral reproduction of strain A/Hong Kong/1/68 (НЗМ2); 7" А1Т9ТИДё(2) - average indicator of inhibition of viral reproduction of strain A/РА/8/34 ( DUMB); ""A1T9TIDvo(3) - the average indicator of exposure to the extracellular virus A/R H/8/34 (NIMI).

Claims (1)

FORMULA OF THE USEFUL MODEL M-Benzylaminomethanesulfonic acid of the formula: / (c) le Her / he (c) as an anti-influenza agent.