RU2469020C1 - (3r,4r,5s)-4-acylamino-5-amino-3-(1-ethyl-propoxy)-cyclohex-1-ene-carboxylic acids, their esters and method for using - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new substituted (3R,4R,5S)-5-amino-4-acylamino-3-(1-ethyl-propoxy)-cyclohex-1-ene carboxylic acids of general formula 1, their esters, pharmaceutically acceptable salts and/or hydrates possessing antiviral activity associated with inhibition of neuraminidase, particularly influenza virus neuraminidase. The invention also refers to pharmaceutical compositions and a based drug preparation, a method for inhibition of neuraminidase, and a method of treating influenza and pneumonia caused by influenza virus. In general formula 1

R1 represents hydrogen, C₁-C₇alkyl, C₂-C₇alkenyl or C₂-C₇alkinyl optionally substituted by C₃-C₆cycloalkyl, phenyl, pyridyl, C₁-C₃alkoxy; R2 represents hydrogen; -NH₂; C₂H₅O-, substituted C₁-C₅alkyl specified in -CH₂OH, -CH₂-NH₂, -CH=CH₂, -CH=CHCl, -C=CH, -CH₂F, -CHF₂, -CF₃, -CH₂CF₃, cyclo-C₃H₇, -CF₂CF₂CF₂CF₃, -CF₂CF₂CF₂CHF₂, -CH₂CF₂CF₂CF₂CF₃.

EFFECT: compounds can find application in treating influenza virus and pneumonia caused by influenza virus.

10 cl, 3 tbl, 33 ex

Description

This invention relates to new compounds - substituted 4,5-diamino-3-alkyloxy-cyclohex-1-ene-carboxylic acids and their esters - inhibitors of neuraminidase activity.

Many of the microorganisms containing neuraminidase are pathogenic to humans and other animals, including birds, horses, pigs and seals. Such pathogens include the influenza virus. Neuraminidase is associated with the pathogenicity of the influenza virus.

In this regard, new substituted 4,5-diamino-3-alkyloxy-cyclohex-1-ene carboxylic acids and their esters are of interest as drug substances for preparations intended for the prevention and treatment of influenza.

The most famous neuraminidase inhibitors are (3R, 4R, 5S) -5-amino-3-alkyloxy-4-acetylamino-cyclohex-1-ene-carboxylic acids of the general formula A1, the most active of which is (3R, 4R, 5S) -5-amino-4-acetylamino-3- (1-ethyl-propoxy) -cyclohex-1-ene-carboxylic acid of the formula A2, which, on the basis of X-ray diffraction data of its complex with influenza virus neuraminidase, effectively binds to the active center of the enzyme ( Oseltamivir Carboxylate) [C.U. Kim, W.Lew, MAWilliams, et al. J. Am. Chem. Soc. 1997, 119, 681-690.].

Oseltamivir carboxylate A3 ethyl ester, known as oseltamivir phosphate or Tamiflu (Oseltamivir Phosphate, Tamiflu) [J.C. Rohloff, K. M. Kent, M. J. Postich, et al. J. Org. Chem. 1998, 63, 4545.], is a precursor of acid A2 and a well-known anti-influenza drug [https://integrity.thomson-pharma.com/].

R = H, C ₁ -C ₇ alkyl

In the source RU 2181357 (publication of international application WO 1996026933) compounds of the general formula A are used

where, inter alia, (a) E1 is —CO ₂ H; G1 is —N (H) (C (NH) (NH ₂ )); T1 is —N (H) (Ac); U1 is —CH2OH; (b) E1 is —CO ₂ H; G1 is —NH2; T1 is —N (H) (Ac); U1 is —CH2OH; (c) E1 is —CH ₂ OH, —CH ₂ OTMS, —CO ₂ H, or —CHO; G1 is —N ₃ ; T1 is —N (H) (Ac); and U1 is —CH ₂ OCH ₂ Ph; (d) E1 is —CO ₂ H or —CO ₂ CH ₃ ; G1 is —NH ₃ ; T1 is —N (H) (Ac); and U1 is —CH ₂ OH.

The resulting compounds are used as inhibitors of viral or bacterial neuraminidases, for the treatment or prevention of influenza virus infection. In vitro studies were carried out according to the described test methods (e.g. Warner and O'Brien, Biochemistry, vol 18, No. 13, pp. 2783-2787, 1979). The authors of RU 2181357 conducted tests on diluted solutions of viruses, and activity was measured by fluorescence of the solution. The experimental data of inhibitory activity are shown in Table 1.

Table 1. Determination of enzymatic activity according to RU No. 2181357. Compound pc50 A2 +++ A4 / A5 (2: 1) ++ A6 ++ A7 ++ A8 ++ A9 ++ A10 + A11 / A12 (1.2: 1) + A13 +++ A14 +++ A15 +++

PK ₅₀ — concentration of the compound at which 50% inhibition of enzymatic activity is observed (+ corresponds to PK ₅₀ = 10–100 µmol, ++ 1–10 µmol, +++ <1.0 µmol).

When carrying out the invention described in RU No. 2181357, in the examples of the invention, the value of T1 is predominantly —NHC (O) CH ₃ , and the preparation and properties of compounds with the value T1 —NHC (O) CH ₂ F, - NHC (O) CHF ₂ , -NHC (O) CF ₃ .

The search for highly effective anti-influenza drugs with increased activity and reduced toxicity is still one of the main directions in creating new pharmacological agents for treating influenza. In this regard, the development of new anti-influenza substances, pharmaceutical compositions and drugs, as well as methods for their preparation and use, is relevant.

The following are definitions of terms that are used in the description of this invention.

“Alkenyl” means an aliphatic linear or branched hydrocarbon group containing from 2 to 7 carbon atoms and including at least one carbon-carbon double bond.

Branched means that one or more lower alkyl groups, such as methyl, ethyl or propyl, are attached to a linear alkenyl chain. An alkyl group may have one or more substituents, for example, such as halogen, alkenyloxy, cycloalkyl, cyano, hydroxy, alkoxy, carboxy, alkynyloxy, aralkoxy, aryloxy, aryloxycarbonyl, alkylthio, heteroaralkyloxy, heterocyclyl, heterocyclylalkyloxyalkyloxycarboxyloxycarbonyl

и

независимо друг от друга представляют собой «заместители аминогруппы», значение которых определено в данном разделе, например атом водорода, алкил, арил, аралкил, гетероаралкил, гетероциклил или гетероарил, или

и

вместе с атомом N, с которым они связаны, образуют через

и

4-7-членный гетероциклил или гетероцикленил. Предпочтительными алкильными группами являются метил, трифторметил, циклопропилметил, циклопентилметил, этил, н-пропил, изо-пропил, н-бутил, трет-бутил, н-пентил, 3-пентил, метоксиэтил, карбоксиметил, метоксикарбонилметил, бензилоксикарбонилметил и пиридилметилоксикарбнилметил. Предпочтительными алкенильными группами являются этенил, пропенил, н-бутенил, изо-бутенил, 3-метилбут-2-енил, н-пентенил, и циклогексилбутенил.Where

and

independently, they are “amino substituents”, the meanings of which are defined in this section, for example, a hydrogen atom, alkyl, aryl, aralkyl, heteroalkyl, heterocyclyl or heteroaryl, or

and

together with the N atom with which they are bonded, form through

and

4-7 membered heterocyclyl or heterocyclenyl. Preferred alkyl groups are methyl, trifluoromethyl, cyclopropylmethyl, cyclopentylmethyl, ethyl, n-propyl, iso-propyl, n-butyl, tert-butyl, n-pentyl, 3-pentyl, methoxyethyl, carboxymethyl, methoxycarbonylmethyl, benzyloxycarbonylmethylmethyl and pyridinyl. Preferred alkenyl groups are ethenyl, propenyl, n-butenyl, iso-butenyl, 3-methylbut-2-enyl, n-pentenyl, and cyclohexylbutenyl.

"Alkenyloxy" means an alkenyl-O-group in which alkenyl is defined in this section. Preferred alkenyloxy groups are allyloxy and 3-butenyloxy.

“Alkenyloxyalkyl” means an alkenyl-O-alkyl group in which alkyl and alkenyl are defined in this section.

"Alkyl" means an aliphatic hydrocarbon linear or branched group with 1-12 carbon atoms in the chain. Branched means that the alkyl chain has one or more "lower alkyl" substituents. Alkyl may have one or more identical or different substituents (“alkyl substituents”), including halogen, alkenyloxy, cycloalkyl, aryl, heteroaryl, heterocyclyl, aroyl, cyano, hydroxy, alkoxy, carboxy, alkynyloxy, aralkoxy, aryloxy, aryloxycarbenyl, alkylthio, heteroarylthio, aralkylthio, arylsulfonyl, alkylsulfonylheteroaralkyloxy, annelated heteroaryl ring, alkenyl, annelated heteroarylcycloalkyl, annelated heteroaryl heterocyclenyl, annelated heteroaryl heterocyclyl, annelated aero cycloalkenyl, annelated arylcycloalkyl, annelated arylheterocyclenyl, annelated arylheterocyclyl, alkoxycarbonyl, aralkoxycarbonyl, or geteroaralkiloksikarbonil

где

и

независимо друг от друга представляют собой «заместители аминогруппы», значение которых определено в данном разделе, например атом водорода, алкил, арил, аралкил, гетероаралкил, гетероциклил или гетероарил, или

и

вместе с атомом N, с которым они связаны, образуют через

и

4-7-членный гетероциклил или гетероцикленил. Предпочтительными алкильными группами являются метил, трифторметил, циклопропилметил, циклопентилметил, этил, н-пропил, изо-пропил, н-бутил, трет-бутил, н-пентил, 3-пентил, метоксиэтил, карбоксиметил, метоксикарбонилметил, этоксикарбонилметил, бензилоксикарбонилметил метоксикарбонилметил и пиридилметилоксикарбнилметил. Предпочтительными «алкильными заместителями» являются циклоалкил, арил, гетероарил, гетероциклил, гидрокси, алкокси, алкоксикарбонил, аралкокси, арилокси, алкилтио, гетероарилтио, аралкилтио, алкилсульфонил, арилсульфонил, алкоксикарбонил, аралкоксикарбонил, гетероаралкилоксикарбонил или

аннелированный арилгетероцикленил, аннелированный арилгетероциклил.

Where

and

independently, they are “amino substituents”, the meanings of which are defined in this section, for example, a hydrogen atom, alkyl, aryl, aralkyl, heteroalkyl, heterocyclyl or heteroaryl, or

and

together with the N atom with which they are bonded, form through

and

4-7 membered heterocyclyl or heterocyclenyl. Preferred alkyl groups are methyl, trifluoromethyl, cyclopropylmethyl, cyclopentylmethyl, ethyl, n-propyl, iso-propyl, n-butyl, tert-butyl, n-pentyl, 3-pentyl, methoxyethyl, carboxymethyl, methoxycarbonylmethylmethyloxycarbylmethyloxycarbonylmethyloxycarbonylmethyloxycarbonylmethyloxycarbonylmethyloxycarbonylmethyloxycarbonylmethyloxycarbonylmethylene . Preferred “alkyl substituents” are cycloalkyl, aryl, heteroaryl, heterocyclyl, hydroxy, alkoxy, alkoxycarbonyl, aralkoxy, aryloxy, alkylthio, heteroarylthio, aralkylthio, alkylsulfonyl, arylsulfonyl, alkoxycarbonyl, aralkoxycarbonyl or

annelated arylheterocyclenyl, annelated arylheterocyclyl.

"Alkynyl" means an aliphatic linear or branched hydrocarbon group containing from 2 to 12 carbon atoms and including at least one carbon-carbon triple bond.

где

и

независимо друг от друга представляют собой «заместители аминогруппы», значение которых определено в данном разделе, например атом водорода, алкил, арил, аралкил, гетероаралкил, гетероциклил или гетероарил, или

и

вместе с атомом N, с которым они связаны, образуют через

и

4-7-членный гетероциклил или гетероцикленил. Предпочтительными алкильными группами являются метил, трифторметил, циклопропилметил, циклопентилметил, этил, н-пропил, изо-пропил, н-бутил, трет-бутил, н-пентил, 3-пентил, метоксиэтил, карбоксиметил, метоксикарбнилметил, бензилоксикарбонилметил и пиридилметилоксикарбнилметил. Предпочтительными алкенильными группами являются этенил, пропенил, н-бутенил, изо-бутенил, 3-метилбут-2-енил, н-пентенил, бута-1,3-диин и гекса-1,3,5-триин.Branched means that one or more lower alkyl groups, such as methyl, ethyl or propyl, are attached to the linear alkynyl chain. An alkyl group may have one or more substituents, for example, such as halogen, alkenyloxy, cycloalkyl, cyano, hydroxy, alkoxy, quinloxy, aralkoxy, aryloxy, aryloxycarbonyl, alkylthio, heteroaralkyloxy, heterocyclyl, heterocyclylalkyloxy, alkoxycarbonyl, aalkoxycarbonyl, a

Where

and

independently, they are “amino substituents”, the meanings of which are defined in this section, for example, a hydrogen atom, alkyl, aryl, aralkyl, heteroalkyl, heterocyclyl or heteroaryl, or

and

together with the N atom with which they are bonded, form through

and

4-7 membered heterocyclyl or heterocyclenyl. Preferred alkyl groups are methyl, trifluoromethyl, cyclopropylmethyl, cyclopentylmethyl, ethyl, n-propyl, iso-propyl, n-butyl, tert-butyl, n-pentyl, 3-pentyl, methoxyethyl, carboxymethyl, methoxycarbnylmethyl, benzyloxycarbonylmethylmethyl and pyridinyl. Preferred alkenyl groups are ethenyl, propenyl, n-butenyl, iso-butenyl, 3-methylbut-2-enyl, n-pentenyl, buta-1,3-diine and hexa-1,3,5-triine.

“Alkoxy” means an alkyl-O-group in which alkyl is defined in this section. Preferred alkyloxy groups are methoxy, ethoxy, n-propoxy, iso-propoxy and n-butoxy.

“Aryl” means an aromatic monocyclic or polycyclic system comprising from 6 to 14 carbon atoms, preferably from 6 to 10 carbon atoms. Aryl may contain one or more “cyclic system substituents”, which may be the same or different. Representative aryl groups are phenyl or naphthyl, substituted phenyl or substituted naphthyl. Aryl can be anelated with a non-aromatic ring system or heterocycle.

“Heterocyclyl” means an aromatic or non-aromatic saturated monocyclic or polycyclic system comprising from 3 to 10 carbon atoms, preferably from 5 to 6 carbon atoms, in which one or more carbon atoms are replaced by a heteroatom such as nitrogen, oxygen, sulfur. The prefix "aza", "oxa" or "thia" before heterocyclyl means the presence in the cyclic system of a nitrogen atom, an oxygen atom or a sulfur atom, respectively. Heterocyclyl may have one or more “cyclic system substituents,” which may be the same or different. The nitrogen and sulfur atoms in the heterocyclyl can be oxidized to N-oxide, S-oxide or S-dioxide. Representatives of heterocyclyl are piperidine, pyrrolidine, piperazine, morpholine, thiomorpholine, thiazolidine, 1,4-dioxane, tetrahydrofuran, tetrahydrothiophene, etc.

"Hydrate" means a solvate in which water is a molecule or molecules of a solvent.

“Substituent” means a chemical radical that is attached to a scaffold (fragment), for example, “substituent alkyl”, “substituent of amino group”, “substituent carbamoyl”, “substituent of the cyclic system”, the meanings of which are defined in this section.

"Medicinal substance" (drug substance) (drug substance), drug-substance) means a physiologically active substance of synthetic or other (biotechnological, plant, animal, microbial and other) origin, having pharmacological activity and is the active principle of the pharmaceutical composition used for the production and the manufacture of a drug (agent).

“Medicinal product (preparation)” - a substance (or a mixture of substances in the form of a pharmaceutical composition) in the form of tablets, capsules of injections, ointments and other finished forms, intended to restore, correct or alter the physiological functions in humans and animals, as well as for the treatment and disease prevention, diagnosis, anesthesia, contraception, cosmetology and more.

Neuraminidase (sialidase, acylneuraminyl hydrolase and EC 3.2.1.18) is an enzyme common to animals and a number of microorganisms. It is a glycohydrolase that cleaves alpha-ketosidically linked sialic acids from glycoproteins, glycolipids and oligosaccharides. Many of the microorganisms containing neuraminidase are pathogenic to humans and other animals, including poultry, horses, pigs and seals. Such pathogens include the influenza virus. Neuraminidase is associated with the pathogenicity of the influenza virus. Presumably, it promotes the elution of newly synthesized virions from infected cells and the movement of the virus (due to its hydrolase activity) through the mucus of the respiratory tract.

"Lower alkyl" means a linear or branched alkyl with 1-5 carbon atoms.

A “therapeutic cocktail” is a simultaneously administered combination of two or more drugs with a different mechanism of pharmacological action and aimed at different biological targets involved in the pathogenesis of the disease.

"Pharmaceutical composition" means a composition comprising a compound of formula 1 and at least one of the components selected from the group consisting of pharmaceutically acceptable and pharmacologically compatible excipients, solvents, diluents, carriers, excipients, distributing and perceiving agents, agents delivery, such as preservatives, stabilizers, fillers, grinders, moisturizers, emulsifiers, suspending agents, thickeners, sweeteners, perfumes, flavors, antibacterial agents, fungicides, lubricants, prolonged delivery regulators, the choice and ratio of which depends on the nature and method of administration and dosage. Examples of suspending agents are ethoxylated isostearyl alcohol, polyoxyethylene, sorbitol and sorbitol ether, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, as well as mixtures of these substances. Protection against the action of microorganisms can be provided with a variety of antibacterial and antifungal agents, for example, parabens, chlorobutanol, sorbic acid and the like. The composition may also include isotonic agents, for example, sugars, sodium chloride and the like. The prolonged action of the composition can be achieved using agents that slow down the absorption of the active principle, for example aluminum monostearate and gelatin. Examples of suitable carriers, solvents, diluents and delivery vehicles are water, ethanol, polyalcohols, and also mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Examples of excipients are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate and the like. Examples of grinders and distributors are starch, alginic acid and its salts, silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talc, and high molecular weight polyethylene glycol. The pharmaceutical composition for oral, sublingual, transdermal, intramuscular, intravenous, subcutaneous, local or rectal administration of the active principle, alone or in combination with another active principle, can be administered to animals and humans in a standard administration form, in the form of a mixture with traditional pharmaceutical carriers. Suitable unit dosage forms include oral forms such as tablets, gelatine capsules, pills, powders, granules, chewing gums and oral solutions or suspensions, sublingual and buccal administration forms, aerosols, implants, local, transdermal, subcutaneous, intramuscular, intravenous, intranasal or intraocular administration forms and rectal administration forms.

“Pharmaceutically acceptable salt” means the relatively non-toxic organic and inorganic salts of the acids and bases of the present invention. These salts can be obtained in situ during the synthesis, isolation or purification of the compounds or prepared specially. In particular, base salts can be prepared on the basis of the purified free base of the claimed compound and a suitable organic or inorganic acid. Examples of salts thus obtained are hydrochlorides, hydrobromides, sulfates, bisulfates, phosphates, nitrates, acetates, oxalates, valeriates, oleates, palmitates, stearates, laurates, borates, benzoates, lactates, tosylates, citrates, maleates, fumarates, succinates, tartrates mesylates, malonates, salicylates, propionates, ethanesulfonates, benzenesulfonates, sulfamates and the like (Detailed description of the properties of such salts is given in Berge SM, et al., Pharmaceutical Salts J. Pharm. Sci. 1977, 66: 1-19). Salts of the claimed acids can also be specially prepared by reacting the purified acid with a suitable base, and metal and amine salts can be synthesized. Metal salts include sodium, potassium, calcium, barium, zinc, magnesium, lithium and aluminum salts, the most desirable of which are sodium and potassium salts. Suitable inorganic bases from which metal salts can be obtained are hydroxide, carbonate, sodium bicarbonate and hydride, potassium hydroxide and bicarbonate, potash, lithium hydroxide, calcium hydroxide, magnesium hydroxide, zinc hydroxide. As organic bases from which salts of the claimed acids can be obtained, amines and amino acids are selected that are sufficiently basic to form a stable salt and are suitable for medical use (in particular, they should have low toxicity). Such amines include ammonia, methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, benzylamine, dibenzylamine, dicyclohexylamine, piperazine, ethylpiperidine, tris (hydroxymethyl) aminomethane and the like. In addition, tetraalkylammonium hydroxides, for example, such as choline, tetramethylammonium, tetraethylammonium and the like, can be used for salt formation. As amino acids, the main amino acids can be used - lysine, ornithine and arginine.

The authors found new neuraminidase inhibitors, which are previously unknown (3R, 4R, 5S) -5-amino-4-acylamino-3- (1-ethyl-propoxy) -cyclohex-1-ene-carboxylic acids and their esters of general formula 1 and their pharmaceutically acceptable salts or hydrates,

where R1 is hydrogen, C ₁ -C ₇ alkyl, C ₂ -C ₇ alkenyl or C ₂ -C ₇ alkynyl optionally substituted with C ₃ -C ₆ cycloalkyl, phenyl, pyridyl, C ₁ -C ₃ alkoxy;

R2 is hydrogen; -NH ₂ ; C ₂ H ₅ O-; substituted C ₁ -C ₃ alkyl selected from —CH ₂ OH, —CH ₂ —NH ₂ , —CH = CH ₂ , —CH = CHCl, —CH≡CH, —CH ₂ F, —CHF ₂ , —CF ₃ , -CH ₂ CF ₃ , cyclo-C ₃ H ₇ , -CF ₂ CF ₂ CF ₂ CF ₃ , -CF ₂ CF ₂ CF ₂ CHF ₂ , -CH ₂ CF ₂ CF ₂ CF ₂ CF ₃ .

According to the invention, preferred compounds are acids of general formula 1.1 and their esters of general formula 1.2,

in which R1 represents C ₁ -C ₅ alkyl, C ₃ -C ₅ alkenyl or C ₃ -C ₅ alkynyl, and R2 has the above meaning.

According to this invention, more preferred compounds are

(3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.1,

(3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.2,

(3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.3,

(3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.4,

(3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.5,

(3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.6,

(3R, 4R, 5S) -5-amino-4- (2,2-difluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.7,

(3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-en-carboxylic acid 1.1.8,

(3R, 4E, 5S) -5-amino-4- (3,3,3-trifluoro-propionylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene carboxylic acid 1.1.9,

(3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5-octafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1-en- carboxylic acid 1.1.10,

(3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,5-nonafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1- en-carboxylic acid 1.1.11,

(3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,6,6,6-undecafluorohexanoylamino) -3- (1-ethylpropoxy) - cyclohex-1-en-carboxylic acid 1.1.12,

(3R, 4R, 5S) -5-amino-4- (cyclopropylcarbonyl-amino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.14,

(3R, 4R, 5S) -5-amino-4- (ethoxycarbonyl-amino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.15,

(3R, 4R, 5S) -5-amino-4-ureido-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.16,

(3R, 4R, 5S) -4- (3-chloroprop-2-enoyl amino) -5-amino-3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.17,

ethyl ether (3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.1,

ethyl ether (3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.2,

ethyl ester of (3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.3,

ethyl ester of (3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.4,

ethyl ester of (3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.5,

ethyl ether (3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.6,

ethyl ester of (3R, 4R, 5S) -5-amino-4- (2,2-difluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.7,

ethyl ether (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.8,

ethyl ester of (3R, 4R, 5S) -5-amino-4- (3,3,3-trifluoro-propionylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.9,

ethyl ester (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5-octafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1- en-carboxylic acid 1.2.10,

ethyl ester (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,5-nonafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex- 1-en-carboxylic acid 1.2.11,

ethyl ester (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,6,6,6-undecafluorohexanoylamino) -3- (1-ethylpropoxy ) -cyclohex-1-ene-carboxylic acid 1.1.12,

ethyl ether (3R, 4R, 5S) -5-amino-4- (cyclopropylcarbonylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.14,

ethyl ester of (3R, 4R, 5S) -5-amino-4-ureido-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.15,

ethyl ether (3R, 4R, 5S) -5-amino-4- (ethoxycarbonylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.16,

ethyl ester of (3R, 4R, 5S) -4- (3-chloroprop-2-enoyl-amino) -5-amino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.17,

allyl ether (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3 - (- ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.18,

(3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid prop-2-ynyl ester 1.2. 19.

(3R, 4R, 5S) -5-amino-4-acylamino-3- (1-ethyl-propoxy) cyclohex-1-ene-carboxylic acids and their esters of general formula 1 are obtained by analogy with the synthesis of the known compounds A2 and A3 [M. Morita et al. Bioorg. Med. Chem. Lett. 18, 600-602 (2008). F. Konno et al. Bioorg. Med. Chem. Lett. 18, 1260-1263 (2008)] according to the scheme below,

where R1 and R2 have the above meaning.

The new substituted 4,5-diamino-3-hydroxy-cyclohex-1-ene-carboxylic acids of general formula 1 and their pharmaceutically acceptable salts and / or hydrates are neuraminidase inhibitors and exhibit anti-influenza activity (Table 1).

According to this invention, the new compounds of General formula 1 are a drug source for the preparation of pharmaceutical compositions and formulations for the prevention and treatment of influenza in warm-blooded animals and humans.

The subject of this invention is a pharmaceutical composition comprising, as a drug principle, the compounds of general formula 1 or their pharmaceutically acceptable salts and / or hydrates in a therapeutically effective amount.

Pharmaceutical compositions may include pharmaceutically acceptable excipients. By pharmaceutically acceptable excipients are meant diluents, excipients and / or carriers used in the pharmaceutical field. The pharmaceutical composition, along with the compound of general formula 1 or their pharmaceutically acceptable salt and / or hydrate of the present invention, may include other active substances, including those having anti-influenza activity, provided that they do not cause undesirable effects.

If necessary, the use of the pharmaceutical composition of the present invention in clinical practice, it can be mixed with traditional pharmaceutical carriers.

Carriers used in the pharmaceutical compositions of the present invention are carriers that are used in the pharmaceutical field to produce common forms, including oral agents using binders, lubricants, disintegrants, solvents, diluents, stabilizers, suspending agents, colorless agents flavoring agents of taste; antiseptic agents, solubilizers, stabilizers are used in injection forms; in local forms, bases, diluents, lubricants, antiseptic agents are used.

The subject of this invention is a method for preparing a pharmaceutical composition by mixing with an inert excipient and / or solvent of at least one drug principle of general formula 1 or a pharmaceutically acceptable salt and / or hydrate thereof in a therapeutically effective amount.

The subject of the present invention is also a medicament for treating influenza in the form of tablets, capsules or injections in a pharmaceutically acceptable package intended for the prophylaxis of treating influenza in humans and warm-blooded animals, comprising an influenza drug principle of the general formula 1 or a new pharmaceutical therapeutically effective amount.

The subject of the present invention is also therapeutic cocktails for treating influenza, comprising, as one of the components, a new drug or a new pharmaceutical composition containing, as an active component, at least one compound of general formula 1 or a pharmaceutically acceptable salt and / or hydrate thereof.

A therapeutic cocktail for and treatment of influenza, along with the medicament of the present invention, may include other known preparations for treating influenza or drugs that enhance the patient's immune system.

In accordance with this invention, a method for the prevention and treatment of influenza in animals and humans consists in administering to the patient a new drug, a new pharmaceutical composition or a new therapeutic cocktail.

Medicines can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or locally. The clinical dosage of the agent of general formula 1 in patients can be adjusted depending on the therapeutic efficacy and bioavailability of the active ingredients in the body, their metabolic rate and excretion, as well as depending on the age, sex and stage of the patient’s disease, the daily dose in adults is usually 10 ~ 500 mg, preferably 50 ~ 300 mg, therefore, during preparation from the pharmaceutical composition of the drug of the present invention in the form of dosage units, the above effective dosage must be taken into account, with each dosage unit of the drug containing 10 ~ 500 mg of the drug of general formula 1, preferably 50 ~ 300 mg. can be taken several times during certain periods of time (preferably one to six times).

The subject of the invention is also a method of inhibiting the activity of a neuraminidase in vivo, including an influenza virus neuraminidase, comprising the step of contacting a compound of general formula 1 and a neuraminidase.

The invention is illustrated, but not limited to the following examples.

Example 1. Obtaining (3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.1. 250 mg of (3R, 4R, 5S) -4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-encarboxylic acid 2.2 was dissolved in 15 ml of toluene, 500 mg of formic acid was added and 1 g of molecular sieve 3A and the reaction mixture was refluxed overnight. Then the reaction mass was filtered, the precipitate was washed with ethanol, the mother liquor was evaporated, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Mg ₂ SO ₄ and then evaporated. Purification of the reaction product was carried out by HPLC. Received (3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.1: LSMS m / z 271 (M + 1), C ₁₃ H ₂₂ N ₂ O ₄ .

Example 2. Obtaining (3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.2. To a solution of 1000 mg (2.7 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1, 620 mg ( 3.24 mmol) of N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride, 438 mg (3.24 mmol) of hydroxybenzotriazole and 1150 mg (8.92 mmol) of N, N-diisopropylethylamine in 10 ml of dry THF added 247 mg (3.24 mmol) A) 2-hydroxyacetic acid. The reaction mass was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Mg ₂ SO ₄ and then evaporated. Ethyl ester (3R, 4R, 5S) -4- (2-hydroxy-acetylamino) -5- (tert-butoxycarbonylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 4 (R1 = Et , R2 = CH ₂ OH) with a yield of 93% according to LCMS. The resulting product (100 mg) was stirred in 1 ml of a 5% aqueous solution of lithium hydroxide for 5-10 minutes at room temperature, then the solvent was distilled off, the residue was dissolved in dioxane, filtered, the resulting solution was evaporated and dried in vacuum. Received acid 5 (R ₂ = CH ₂ OH), dissolved in 1 ml of a 3M solution of hydrochloric acid in dioxane. The resulting solution was stirred at room temperature until the reaction was completed. Monitoring the progress of the reaction was carried out by HPLC. The product was purified by HPLC. Received (3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.1.2: LSMS m / z 301 (M +1), C ₁₄ H ₂₄ N ₂ O ₅ .

Example 3. Obtaining (3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.3. To a solution of 500 mg (1.35 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1, 387 mg ( 2.027 mmol) of N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride, 237 mg (1.76 mmol) of hydroxybenzotriazole and 5 ml of dry DMF were added 308 mg (1.76 mmol) of 2- (tert-butoxycarbonylamino) acetic acid. The reaction mass was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Mg ₂ SO ₄ and then evaporated. The resulting compound was used in further reactions without further purification. The yield of crude product was 604 mg. The alkaline hydrolysis of 4 (R ₁ = Et, R2 = CH ₂ NH ₂ ) was carried out as described in Example 2, and the removal of the Boc-protection of compound 5 (R2 = CH ₂ NH ₂ ) was carried out in trifluoroacetic acid. Received (3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid in the form of trifluoroacetate 1.1.3 · CF ₃ CO ₂ H LSMS m / z 300 (M + 1), C ₁₆ H ₂₆ F ₃ N ₃ O ₆ .

Example 4. Preparation of (3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.4. To a solution of (3R, 4R, 5S) ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate in DMF was added 1.5 equivalents of acrylic acid and 1.5 equivalents of TBTU. The reaction mass was stirred at 50 ° C for 12 hours. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% K ₂ CO ₃ and dried in vacuum. Amide 4 (R1 = Et, R2 = CH = CH ₂ ) was purified by HPLC. Alkaline hydrolysis 4 (R1 = Et, R2 = CH = CH ₂ ) was carried out as described in Example 2. Product 5 (R2 = CH = CH ₂ ) (50 mg) was hydrolyzed in 0.5 ml of dioxane and 50 μl of sulfuric acid. The reaction malissa was stirred at room temperature until the reaction was completed. Monitoring the progress of the reaction was carried out by LCMS. At the end of the reaction, sodium bicarbonate was added to the solution until the sulfuric acid was completely neutralized, then the solvent was distilled off and isolated using HPLC (3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) -cyclohex-1- en-carboxylic acid 1.1.4: LSMS m / z 297 (M + 1), C ₁₅ H ₂₄ N ₂ O ₄ . In a similar manner, (3R, 4R, 5S) -4- (3-chloroprop-2-enoyl amino) -5-amino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid was obtained 1.1.17: LSMS m / z 331 (M + 1), C ₁₅ H ₂₃ ClN ₂ O ₄ .

Example 5. Obtaining (3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.1.5. To a solution of 750 mg (2 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate in 7.5 ml of methylene chloride 213 mg (3.04 mmol) of propionic acid and 626 mg (3.05 mmol) of N, N'-dicyclohexylcarbodiimide were added. The resulting mixture was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ether and the precipitate was filtered off. The mother liquor was evaporated and the obtained compound was used in further reactions without further purification. The yield of crude product was 61%. Alkaline hydrolysis 4 (R1 = Et, R2 = С≡СН) was carried out as described in Example 2. Product 5 (R2 = С≡СН) (50 mg) was hydrolyzed in 0.5 ml of dioxane and 50 μl of sulfuric acid. The reaction mass was stirred at room temperature until the reaction was completed. Monitoring the progress of the reaction was carried out by LCMS. At the end of the reaction, sodium bicarbonate was added to the solution until the sulfuric acid was completely neutralized, then the solvent was distilled off and isolated using HPLC (3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) -cyclohex-1- en-carboxylic acid 1.1.5: LSMS m / z 295 (M + 1), C ₁₅ H ₂₂ N ₂ O ₄ .

Example 6. Obtaining (3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.1.6 was carried out by analogy with the synthesis acids 1.1.4, as described in example 4. LSMS m / z 303 (M + 1), C ₁₄ H ₂₃ FN ₂ O ₄ .

Example 7. The preparation of (3R, 4R, 5S) -5-amino-4- (2,2-difluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.7 was carried out by analogy with the synthesis of acid 1.1.4, as described in example 4. LSMS m / z 321 (M + 1), C ₁₄ H ₂₂ F ₂ N ₂ O ₄ .

Example 8. The preparation of (3K, 4K, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.8 was carried out by analogy with the synthesis of acid 1.1.4, as described in example 4. LSMS m / z 339 (M + 1), C ₁₄ H ₂₁ F ₃ N ₂ O ₄ .

Example 9. Preparation of (3R, 4R, 5S) -5-amino-4- (3,3,3-trifluoro-propionylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.9. To a solution of 300 mg (0.81 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1, 186 mg ( 0.972 mmol) of N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride, 132 mg (0.972 mmol) of hydroxybenzotriazole and 345 mg (2.68 mmol) of N, N-diisopropylethylamine in 10 ml of dry THF were added 126 mg (0.972 mmol) ) 3,3,3-trifluoropropionic acid. The reaction mass was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Na ₂ SO ₄ and then evaporated. The product yield according to the LCMS spectrum was 80%. The resulting compound was used in further reactions without further purification. Alkaline hydrolysis 4 (R1 = Et, R2 = CH ₂ CF ₃ ) and acid hydrolysis 5 (R ₂ = CH ₂ CF ₃ ) were carried out as described in example 2. Received (3R, 4R, 5S) -5-amino-4 - (3,3,3-trifluoro-propionylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.1.9: LSMS m / z 353 (M + 1), C ₁₅ H ₂₃ F ₃ N ₂ O ₄ .

Example 10. Obtaining (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,4-heptafluorobutyrylamino) -3- (1-ethylpropoxy) cyclohex-1-en- carboxylic acid 1.1.10. To a solution of 150 mg (0.4 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-encarboxylate 2.1 and 113 mg ( 1.22 mmol) of triethylamine in 1.5 ml of dry dichloromethane was added 142 mg (0.6 mmol) of the acid chloride of 2,2,3,3,4,4,4-heptafluorobutanoic acid. The reaction mass was stirred at room temperature until the reaction was completed. Monitoring the progress of the reaction was carried out by LCMS. At the end of the reaction, the obtained compound was used in further reactions without further purification. The yield of crude product is close to quantitative. Alkaline hydrolysis 4 (R1 = Et, R2 = CF ₂ CF ₂ CF ₃ ) and acid hydrolysis 5 (R2 = CF ₂ CF ₂ CF ₃ ) were carried out as described in example 2. Received (3R, 4R, 5S) -5- amino-4- (2,2,3,3,4,4,4-heptafluoro-butyrylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.10: LSMS m / z 439 ( M + 1), C ₁₆ H ₂₁ F ₇ N ₂ O ₄ .

Example 11. Obtaining (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5-octafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex- 1-en-carboxylic acid 1.1.11. To a solution of 150 mg (0.4 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-encarboxylate 2.1 and 113 mg ( 1.22 mmol) of triethylamine in 1.5 ml of dry dichloromethane was added 117 mg (0.45 mmol) of acid chloride 2,2,3,3,4,4,5,5-octafluoropentanoic acid. The reaction mass was stirred at room temperature until the reaction was completed. Monitoring the progress of the reaction was carried out by LCMS. At the end of the reaction, the obtained compound was used in further reactions without further purification. The yield of crude product is close to quantitative. Alkaline hydrolysis 4 (R1 = Et, R2 = CF ₂ CF ₂ CF ₂ CHF ₂ ) and acid hydrolysis 5 (R2 = CF ₂ CF ₂ CF ₂ CHF ₂ ) were carried out as described in example 2. Received (3R, 4R, 5S ) -5-amino-4- (2,2,3,3,4,4,5,5-octafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.11: LSMS m / z 471 (M + 1), C ₁₇ H ₂₂ F ₈ N ₂ O ₄ .

Example 12. Obtaining (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,6,6,6-undecafluorohexanoylamino) -3- (1 ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.12. To a solution of 180 mg (0.486 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1 and 148 mg ( 1.46 mmol) of triethylamine in 1.8 ml of dry dichloromethane was added 194 mg (0.73 mmol) of acid chloride 2,2,3,3,4,4,5,5,5-nonafluoropentanoic acid. The reaction mass was stirred at room temperature until the reaction was completed. Monitoring the progress of the reaction was carried out by LCMS. At the end of the reaction, the obtained compound was used in further reactions without further purification. The yield of crude product is close to quantitative. Alkaline hydrolysis 4 (R1 = Et, R2 = CF ₂ CF ₂ CF ₂ CF ₃ ) and acid hydrolysis 5 (R2 = CF ₂ CF ₂ CF ₂ CF ₃ ) were performed as described in example 2. Received (3R, 4R, 5S ) -5-amino-4- (2,2,3,3,4,4,5,5,6,6,6,6-undecafluorohexanoylamino) -3- (1-ethylpropoxy) cyclohex-1-en- carboxylic acid 1.1.12: LSMS m / z 489 (M + 1), C ₁₇ H ₂₁ F ₉ N ₂ O ₄ .

Example 13. The preparation of (3R, 4R, 5S) -5-amino-4- (cyclopropylcarbonylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.14 was carried out by analogy with the synthesis of acid 1.1.4 as described in example 4. LSMS m / z 311 (M + 1), C ₁₆ H ₂₆ N ₂ O ₄ .

Example 14. Obtaining (3R, 4R, 5S) -5-amino-4- (ethoxycarbonylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.15. To a solution of 300 mg (0.81 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate and 157 mg (1.2 mmol) N, N-diisopropylethylamine in 3 ml of dichloromethane was added 114 mg of ethyl chloroformate. The reaction mass was stirred at room temperature for 1 hour. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with water, dried over Na ₂ SO ₄ , and evaporated. The resulting compound was used in further reactions without further purification. Alkaline hydrolysis 4 (R1 = Et, R2 = OEt) and acid hydrolysis 5 (R2 = OEt) were performed as described in Example 2. (3R, 4R, 5S) -5-amino-4- (ethoxycarbonylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.1.15: LSMS m / z 315 (M + 1), C ₁₅ H ₂₆ N ₂ O ₅ .

Example 15. Obtaining (3R, 4R, 5S) -5-amino-4-ureido-3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.1.16. To a solution of 90 mg (0.243 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-encarboxylate 2.1 in 900 μl of ice-cold acetic acid was added 22 mg (0.267 mmol) of KNCO. The reaction mass was stirred at room temperature for 1 hour. Then, the solvent was distilled off, and the obtained compound was used in further reactions without further purification. Alkaline hydrolysis 4 (R1 = Et, R2 = NH ₂ ) and acid hydrolysis 5 (R2 = NH ₂ ) was carried out as described in Example 2. (3R, 4R, 5S) -5-amino-4-ureido-3- was obtained (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.1.16: LSMS m / z 286 (M + 1), C ₁₃ H ₂₃ N ₃ O ₄ .

Example 16. Obtaining ethyl ester hydrochloride of (3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.1. To a solution of 750 mg (2 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1 in 15 ml toluene was added 242 mg (5.26 mmol) of formic acid and 1 g of molecular sieves 3A. The reaction mass was refluxed overnight. Then the reaction mass was filtered, the mother liquor was washed with 5% soda solution, dried over Mg ₂ SO ₄ and then evaporated. Purification of the reaction product was carried out by HPLC. 262 mg of product 4 was obtained (R1 = Et, R2 = H), Boc protection was removed with hydrochloric acid, purified by HPLC and transferred to hydrochloride. Received (3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.1 · HCl: LSMS m / z 299 (M + 1), C ₁₅ H ₂₇ ClN ₃ O ₄ .

Example 17. Preparation of (3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.2. To a solution of 1000 mg (2.7 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1, 620 mg ( 3.24 mmol) N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride, 438 mg (3.24 mmol) of hydroxybenzotriazole and 1150 mg (8.92 mmol) of N, N-diisopropylethylamine in 10 ml of dry THF were added 247 mg (3.24 mmol) ) 2-hydroxyacetic acid. The reaction mass was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Mg ₂ SO ₄ and then evaporated. The product yield according to the LCMS spectrum was 93%. The resulting product 4 (R1 = Et, R2 = CH ₂ OH) was used in further reactions without further purification, Boc protection was removed with hydrochloric acid, purified by HPLC and transferred to hydrochloride. Received 180 mg of ethyl ester hydrochloride (3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.2.2 · HCl: LSMS m / z 329 (M + 1), C ₁₆ H ₂₈ ClN ₂ O ₅ .

Example 18. Preparation of (3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl dihydrochloride 1.2.3. To a solution of 500 mg (1.35 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1, 387 mg ( 2.027 mmol) of N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride, 237 mg (1.76 mmol) of hydroxybenzotriazole and 5 ml of dry DMF were added 308 mg (1.76 mmol) of 2- (tert-butoxycarbonylamino) acetic acid. The reaction mass was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Mg ₂ SO ₄ and then evaporated. The yield of crude product was 604 mg. The resulting product 4 (R1 = Et, R2 = CH ₂ NH ₂ ) was used in further reactions without further purification, Boc protection was removed with hydrochloric acid, purified by HPLC and transferred to dihydrochloride. Received 180 mg of ethyl ester dihydrochloride (3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.2.3 · 2HCl: LSMS m / z 328 (M + 1), C ₁₆ H ₃₁ C ₁₂ N ₃ O ₄ .

Example 19 Preparation of (3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.4 · HCl. To a solution of (3R, 4R, 5S) ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate in DMF was added 1.5 equivalents of acrylic acid and 1.5 equivalents of TBTU. The reaction mass was stirred at 50 ° C for 12 hours. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% K ₂ CO ₃ and dried in vacuum. Purification of amide 4 (R1 = Et, R ₂ = CH = CH ₂ ) was carried out by HPLC. From the resulting product 4 (R1 = Et, R2 = CH ₂ NH ₂ ), the Boc protection was removed with hydrochloric acid, purified by HPLC and transferred to hydrochloride. (3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.4 · HCl: LSMS m / z 325 (M + 1), C ₁₇ H ₂₉ ClN ₂ O ₄ . In a similar manner, (3R, 4R, 5S) -4- (3-chloroprop-2-enoyl-amino) -5-amino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2 was obtained. 17 HCl: LSMS m / z 359 (M + 1), C ₁₇ H ₂₇ ClN ₂ O ₄ .

Example 20. Obtaining phosphate and hydrochloride of ethyl ether (3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.2.5. To a solution of 750 mg (2 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamine) -3- (pentan-3-yloxy) cyclohex-1-encarboxylate 2.1 in 7.5 ml of chloride 213 mg (3.04 mmol) of propionic acid and 626 mg (3.05 mmol) of N, N'-dicyclohexylcarbodiimide were added to methylene. The resulting mixture was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ether and the precipitate was filtered off. The mother liquor was evaporated and the obtained compound was used in further reactions without further purification. The yield of crude product 4 (R1 = Et, R2 = C≡CH) in the form of hydrochloride was 61%, Boc protection was removed with hydrochloric acid, purified using HPLC and converted to phosphate. 203 mg of (3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester phosphate was obtained, which was converted to 1.2.5 · HCl: LSMS hydrochloride m / z 323 (M + 1), C ₁₇ H ₂₇ ClN ₂ O ₄ .

Example 21. Preparation of (3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.6 · HCl carried out by analogy with 1.2.4 · HCl, as described in example 19. Received the ethyl ester hydrochloride (3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) - cyclohex-1-ene-carboxylic acid 1.2.6 HCl: LSMS m / z 331 (M + 1), C ₁₆ H ₂₈ ClFN ₂ O ₄ .

Example 22 Preparation of (3R, 4R, 5S) -5-amino-4- (2,2-difluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.7 . To a solution of 300 mg (0.81 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1, 186 mg ( 0.972 mmol) of N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride, 132 mg (0.972 mmol) of hydroxybenzotriazole and 345 mg (2.68 mmol) of N, N-diisopropylethylamine in 10 ml of dry THF were added 93 mg (0.972 mmol) ) 2,2-difluoroacetic acid. The reaction mass was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Na ₂ SO ₄ and then evaporated. The yield of crude product 4 (R1 = Et, R2 = CHF ₂ ) was 307 mg. The resulting compound was hydrolyzed without further purification in 1 ml of trifluoroacetic acid. The reaction mass was stirred at room temperature for 30 minutes. Then, the solvent was distilled off and purified using HPLC. Received (3R, 4R, 5S) -5-amino-4- (2,2-difluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.7 · HCl: LSMS m / z 349 (M + 1), C ₁₆ H ₂₇ ClF ₂ N ₂ O ₄ .

Example 23. Obtaining ethyl ester hydrochloride of (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.2 .8. To a solution of 300 mg (0.8 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1 and 158 mg ( 1.22 mmol) of N, N-diisopropylethylamine in 3.6 ml of dry dichloromethane was added 388 mg (1.46 mmol) of 2,2,3,3,4,4,5,5,5-non-naphthoropentanoic acid chloride. The reaction mass was stirred at room temperature until the reaction was completed. Monitoring the progress of the reaction was carried out by LCMS. At the end of the reaction, the obtained compound was used in further reactions without further purification. The yield of crude product 4 (R1 = Et, R2 = CF ₃ ) is close to quantitative. The resulting compound was hydrolyzed without further purification in 1 ml of trifluoroacetic acid. The reaction mass was stirred at room temperature for 30 minutes. Then, the solvent was distilled off. The resulting product was subjected to HPLC. (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid ethyl ester hydrochloride 1.2.8- HCl: LS MS m / z 367 (M + 1), C ₁₆ H ₂₆ ClF ₃ N ₂ O ₄ .

Using starting compound 2.1 (3R, 4R, 5S) -allyl-4-amino-5- (tert-butoxycarbonylamine) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate and (3R, 4R, 5S) -prop-2-ynyl-4-amino-5- (tert-butoxycarbonylamine) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate similarly obtained allyl ether hydrochloride (3R, 4R, 5S) -5-amino 4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.18 · HCl: LSMS m / z 379 (M + 1), C ₁₇ H ₂₆ ClF ₃ N ₂ O ₄ and (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex prop-2-ynyl ester hydrochloride -1-en-carboxylic acid 1.2.19 HCl: LSMS m / z 377 (M + 1), C ₁₇ H ₂₄ ClF ₃ N ₂ O ₄ .

Using as the starting compound the corresponding (3R, 4R, 5S) -4-amino-5- (tert-butoxycarbonylamine) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1,

(3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid hydrochloride of 2-cyclohexyl ethyl ester 1.2.20 HCl: LS MS m / z 449 (M + 1), C ₂₂ H ₃₆ ClF ₃ N ₂ O ₄ ;

(3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid hydrochloride 1.2.21 HCl: LS MS m / z 443 (M + 1), C ₂₂ H ₃₀ ClF ₃ N ₂ O ₄ ;

2-pyridin-3-yl-ethyl ester hydrochloride (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-en-carboxylic acids 1.2.22 HCl: LS MS m / z 444 (M + 1), C ₂₁ H ₂₉ ClF ₃ N ₃ O ₄ ;

(3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene carboxylic acid hydrochloride 1.2.23 HCl: LS MS m / z 397 (M + 1), C ₁₇ H ₂₈ ClF ₃ N ₂ O ₅ .

Example 24. Obtaining ethyl ester hydrochlorides of (3R, 4R, 5S) -5-amino-4- (acylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.9 · HCl-1.2.12 · HCl. To a solution of 300 mg (0.81 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamine) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1, 186 mg ( 0.972 mmol) of N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride, 132 mg (0.972 mmol) of hydroxybenzotriazole and 345 mg (2.68 mmol) of N, N-diisopropylethylamine in 10 ml of dry THF were added 126 mg (0.972 mmol) ) 3,3,3-trifluoropropionic acid. The reaction mass was stirred at room temperature overnight. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with 5% soda solution, dried over Na ₂ SO ₄ and then evaporated. The product yield according to the LCMS spectrum was 80%. The resulting compound 4 (R1 = Et, R2 = CH ₂ CF ₃ ) was hydrolyzed without further purification in 1 ml of trifluoroacetic acid. The reaction mass was stirred at room temperature for 30 minutes. Then, the solvent was distilled off. The resulting product was subjected to HPLC. Received the ethyl ester hydrochloride of (3R, 4R, 5S) -5-amino-4- (3,3,3-trifluoro-propionylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.9 HCl: LSMS m / z 381 (M + 1), C ₁₆ H ₂₆ ClF ₃ N ₂ O ₄ .

Similarly received the following.

(3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,4-tert-butyrylamino) -3- (1-ethylpropoxy) cyclohex-1-ene ethyl ester hydrochloride -carboxylic acid 1.2.10 HCl: LSMS m / z 467 (M + 1), C ₁₈ H ₂₆ ClF ₇ N ₂ O ₄ .

(3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5-octafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1 ethyl ester hydrochloride en-carboxylic acid 1.2.11 HCl: LSMS m / z 499 (M + 1), C ₁₉ H ₂₇ ClF ₈ N ₂ O ₄ .

Ethyl ester hydrochloride (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,6,6,6-undecafluorohexanoylamino) -3- (1- ethyl propoxy) cyclohex-1-ene-carboxylic acid 1.2.12 HCl: LSMS m / z 517 (M + 1), C ₁₉ H ₂₆ ClF ₉ N ₂ O ₄ .

Example 25. Obtaining ethyl ester hydrochloride (3R, 4R, 5S) -5-amino-4- (cyclopropylcarbonyl-amino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.2.14 · HCl was carried out according to analogy with 1.2.4 · HCl, as described in example 19. Received the ethyl ester hydrochloride (3R, 4R, 5S) -5-amino-4- (cyclopropylcarbonyl-amino) -3- (1-ethylpropoxy) -cyclohex-1- en-carboxylic acid 1.2.14 HCl: LSMS m / z 339 (M + 1), C ₁₈ H ₃₁ ClN ₂ O ₄ .

Example 26. Preparation of (3R, 4R, 5S) -5-amino-4-ureido-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.15 · HCl. To a solution of 90 mg (0.243 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamino) -3- (pentan-3-yloxy) cyclohex-1-encarboxylate 2.1 in 900 μl of ice-cold acetic acid was added 22 mg (0.267 mmol) of KNCO. The reaction mass was stirred at room temperature for 1 hour. Then, the solvent was distilled off, and compound 4 (R1 = Et, R2 = NH ₂ ) was obtained, Boc protection was removed with hydrochloric acid, purified by HPLC to obtain ethyl ester hydrochloride (3R, 4R, 5S) -5-amino-4-ureido- 3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.2.15 HCl: LSMS m / z 343 (M + 1), C ₁₇ H ₃₁ ClN ₂ O ₅ .

Example 27. Preparation of (3R, 4R, 5S) -5-amino-4- (ethoxycarbonyl-amino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.16 · HCl. To a solution of 300 mg (0.81 mmol) of the starting (3R, 4R, 5S) -ethyl-4-amino-5- (tert-butoxycarbonylamine) -3- (pentan-3-yloxy) cyclohex-1-enecarboxylate 2.1 and 157 mg ( 1.2 mmol) N, N-diisopropylethylamine in 3 ml of dichloromethane was added 114 mg of ethyl chloroformate. The reaction mass was stirred at room temperature for 1 hour. At the end of the reaction, the solvent was distilled off, the dry residue was dissolved in ethyl acetate, washed with water, dried over Na ₂ SO ₄ , and evaporated. From the obtained compound 4 (R1 = Et, R2 = OEt), the Boc-protection was removed with hydrochloric acid, purified by HPLC to obtain 198 mg of (3R, 4R, 5S) -5-amino-4- (ethoxycarbonyl-amino) ethyl hydrochloride -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.2.16 · HCl: LSMS m / z 314 (M + 1), C ₁₅ H ₂₈ ClN ₃ O ₄ .

Example 28. Obtaining a drug in the form of tablets. 1600 mg of starch, 1600 mg of ground lactose, 400 mg of talc and 1000 mg of ethyl ether (3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) -cyclohex-1-ene-carbon were mixed acids 1.2.1 HCl. The resulting bar was crushed into granules and sieved through sieves, collecting 14-16 mesh granules. The granules obtained were tabletted into a suitable tablet form weighing 560 mg each.

Example 29. Preparation of the drug in capsule form. (3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene-carboxylic acid 1.2.2 · HCl mixed thoroughly with powder lactose in a ratio of 2: 1. The resulting powdery mixture was packaged in 300 mg in a suitable size gelatin capsule.

Example 30. Obtaining a medicinal product in the form of injection compositions for intramuscular, intraperitoneal or subcutaneous injection. 500 mg of (3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) -cyclohex-1-ene carboxylic acid ethyl ester hydrochloride 1.2.6 · HCl with 300 mg of chlorobutanol, 2 ml of propylene glycol and 100 ml of injection water. The resulting solution was filtered and placed in 1 ml ampoules, which were then sealed.

Example 31. Determination of the anti-influenza activity of the compounds of the general formula 1. The anti-influenza activity of the preparations in vitro was determined on cultures of MDCK cells infected with the H1N1 influenza virus (infection multiplicity ~ 100 CCID ₅₀ / well). Test drugs were dissolved in DMSO (20 mg / ml) and stored at -20 ° C. Before use, eight 3.3-fold dilutions of the test drugs (100 to 0.03 μg / ml) were prepared in MEM medium containing 50 mg / ml gentamicin and a 0.25% trypsin / EDTA solution. Each dilution was added to 5 wells with a monolayer of MDCK cells (96-well plates). Three out of every 5 wells were infected with strain A influenza virus (isolate California / 07/2009), the remaining two wells were used to determine the cytotoxicity of the preparations. Ribavirin and Oseltamivir carboxylate were used as standard inhibitors. Cell cultures were incubated for 3-4 days until the maximum cytopathic effect was achieved in infected wells containing no inhibitors. After this, the cultures were stained with neutral red for two hours, extracted with a mixture of equal volumes of Sorensen citrate buffer and ethanol, and the absorbance in the wells was read using a spectrophotometer. The concentration of the drug, reducing the cytopathic effect of the virus by 50%, was taken as an inhibitory concentration of 50 (EC ₅₀ ). The concentration of the drug, which reduces the survival of uninfected cells by 50%, was taken as the cytotoxic concentration of 50 (CC ₅₀ ). Table 1 presents data on activity (EC ₅₀ ) and toxicity (CC ₅₀ ) for some new compounds of general formula 1, confirming their high anti-influenza activity: + corresponds to EC ₅₀ > 10 µM, ++ corresponds to EC ₅₀ from 1 to 10 µM and +++ corresponds to EC ₅₀ <1 µM.

Most of the new compounds of general formula 1 show a higher anti-influenza activity than the control compound at the same toxicity level.

Table 2. Influenza activity (EC ₅₀ ) and toxicity (CC ₅₀ ) of compounds of the general formula 1 in vitro on MDCK cell cultures infected with the H1N1 influenza virus Connections EU ₅₀ SS ₅₀ , tsM A3 (Tamiflu Control) ++ (1.5 µM) > 100 1.1.1 +++ > 100 1.1.2 +++ > 100 1.1.3 ++ > 100 1.1.4 +++ > 100 1.1.5 +++ > 100 1.1.6 ++ > 100 1.1.7 +++ > 100 1.1.8 +++ > 100 1.1.9 +++ > 100 1.1.10 ++ > 100 1.1.11 ++ > 100 1.1.12 ++ > 100 1.1.14 +++ > 100 1.1.15 +++ > 100

Table 2. 1.1.16 +++ > 100 1.1.17 +++ > 100 1.2.1 +++ > 100 1.2.2 +++ > 100 1.2.3 ++ > 100 1.2.4 +++ > 100 1.2.5 +++ > 100 1.2.6 ++ > 100 1.2.7 +++ > 100 1.2.8 +++ > 100 1.2.9 +++ > 100 1.2.10 ++ > 100 1.2.11 ++ > 100 1.2.12 ++ > 100 1.2.14 +++ > 100 1.2.15 +++ > 100 1.2.16 +++ > 100 1.2.17 +++ > 100 1.2.18 ++ > 100 1.2.19 ++ > 100

Most of the new compounds of the general formula 1 show a higher anti-influenza activity compared to the control compound and are on average 1000 times higher than the activity of the compounds from the source RU 2181357 (Table 1).

Example 32. Determination of the activity of the compounds of general formula 1 in relation to the neuraminidase of the allantoic influenza virus A / California / 07/09 (H1N1) in accordance with the method of [WHO COLLABORATING CENTER FOR REFERENCE & RESEARCH ON INFLUENZA, AUSTRALIA, Standard Operating Procedure WHO - 025. Reviewed by: Aeron Hurt, Senior Scientist Review Date: 13/3/2009]. In preliminary experiments, working dilutions for the strain of the allantoic influenza virus A / California / 07/09 (H1N1) were determined. For this, 60 μl of a double-diluted virus was prepared in the reaction buffer mixture (RBS, 50 mM MES, 5 mM CaCl ₂ , pH 6.5) in a 96-well round-bottom plate. From this plate, 50 μl of the double-diluted virus was transferred to 96-well flat-bottom plates for measuring fluorescence (FluoroNunc, black, Cat. No. 237105), then an equal volume of substrate buffer was added to them (SB, 12.5 mM 2 '- (4-methylumbelliferyl) -α-DN-acetylneuraminic acid, Sigma, in 40 mM acetate buffer pH 5.8). Wells were used as a control, to which 50 μl of RBS was added instead of the virus. After the plate was incubated at 37 ° for 1 hour, with gentle swaying, 100 μl of stop solution (2.225 mL 0.824 M NaOH in 11.0 mL ethanol) was added to each well of the tablet, after which the fluorescence was measured on a Varioskan Flach instrument ( Thermo Scientific) at λ _ex = 360 nm and λ _em = 448 nm. For further work, those virus dilutions were selected that corresponded to the middle of the linear portion of the curve of the virus dilution versus fluorescence value. To determine the antineuraminidase activity of compounds in 96-well flat-bottom plates for measuring fluorescence (FluoroNunc, black, Cat. No.237105), 50 μl of dilutions of the compounds, Oseltamivir carboxylate, Zanamivir (50 ml) were added to the RBS wells concentration of 0.03; 0.3; 3; 30; 300; 3000; 30000 nm - each concentration per row). As a viral control, wells of row A were used, to which 50 μl of RBS was added. Next, 50 μl of the selected working dilutions of each virus on RBS was added to the corresponding wells. As a control, wells were used to which the same volume of RBS was added instead of the virus. After stirring and incubation at room temperature for 45 minutes, an equal volume of SB was added to all wells. After mixing and incubating the plate at 37 ° C for 1 hour, 100 μl of stop solution was added to each well of the plate. Fluorescence was measured using a Varioskan Flach instrument (Thermo Scientific) at λ _ex = 360 nm and λ _em = 448 nm. All determinations were carried out in at least two replicates (two wells of the plate). The percentage inhibition of neuraminidase activity by the studied substance of the general formula 1 was determined by the formula: percentage inhibition = 100- (UEF experience - UEF control / UEF virus control in the absence of compound - UEF control). The concentration of the drug, reducing the value of the UEF by 50%, was taken as the inhibitory concentration of 50 (IC ₅₀ ). Table 2 presents the activity data (IC ₅₀ ) for some of the new compounds of general formula 1, confirming their high activity against neuraminidase of the allantoic virus A / California / 07/09 (H1N1): + corresponds to an IC ₅₀ > 10 nM , ++ corresponds to an IR ₅₀ of 1 to 10 nM and +++ corresponds to an IR ₅₀ <1 nM.

Table 3 The activity (IC ₅₀ ) of the compounds of general formula 1 with respect to the neuraminidase of the allantoic virus A / California / 07/09 (H1N1). Connections IR ₅₀ Oseltamivir carboxylate A2 (Control) ++ 1.1.1 +++ 1.1.2 ++ 1.1.3 ++ 1.1.4 ++ 1.1.5 ++ 1.1.6 +++ 1.1.7 +++ 1.1.8 +++ 1.1.9 +++ 1.1.10 ++ 1.1.11 ++ 1.1.12 ++ 1.1.14 +++ 1.1.15 ++ 1.1.16 +++ 1.1.17 +++ 1.2.1 +++ 1.2.2 ++ 1.2.3 ++ 1.2.4 ++ 1.2.5 ++ 1.2.6 +++ 1.2.7 +++ 1.2.8 +++ 1.2.9 +++

Table 3. 1.2.10 ++ 1.2.11 ++ 1.2.12 ++ 1.2.14 +++ 1.2.15 +++ 1.2.16 +++ 1.2.17 +++ 1.2.18 ++ 1.2.19 ++

Example 33. The study of the influenza activity of the compounds of General formula 1 in a model of influenza pneumonia in mice. Pre-weighed mice (non-linear females, average weight 12-15 g) were infected intranasally under mild ether anesthesia with the influenza virus A / Aichi / 2/69 (H3N2) (10 LD ₅₀ in 50 μl). In a preliminary experiment, LD ₅₀ was determined by titration, an allantoic virus in the same mice, which were then used in the main experiment. The following treatment regimen for the compounds was used: 24 hours before infection, 1 hour before infection, 24 hours later, and then 1 time per day for 5 days. For oral administration, a disposable insulin syringe with a special needle (lavage) was used, the effect of the following doses was studied: 25 mg / kg / day of compounds in a volume of 100 μl. As a comparison drug, Tamiflu was used at a dose of 25 mg / kg / day. In the group of “virus control” there were 10 animals, in the groups “treated with compounds” of general formula 1 (1.2.1 · HCl, 1.2.2 · HCl, 1.2.5 · HCl, 1.2.8 · HCl, 1.2.9 · HCl) and Tamiflu had 10 mice each. The treated and control animals were monitored daily; in the first 5 days after infection, mice were weighed every day, then every other day. The chemotherapeutic activity of the compounds in the mouse influenza pneumonia model was evaluated by the protection against fatal viral infection, and weight loss in the animal groups treated with the drug, compared with the control group. The decrease or increase in weight was calculated separately for each mouse and expressed as a percentage. Moreover, the weight of the animal before infection was taken as 100%. For all mice of the same group, the average percentage loss or weight gain was determined.

In a preliminary experiment, the dose of the virus is determined containing 10 LD ₅₀ in a volume of 100 μl. All animals in the experiment are infected with this dose of the virus. The efficacy of compounds of the general formula 1 (1.2.1 · HCl, 1.2.2 · HCl, 1.2.5 · HCl, 1.2.8 · HCl, 1.2.9 · HCl) in the mouse influenza pneumonia model was evaluated by the number of animals surviving after infection virus, life expectancy, and weight change in infected animals.

It was found that on the 7th day of observation, all mice infected with the virus and not treated with these compounds (group "virus control") died.

The experiments showed that by the last day of the death of animals from the group of "viral control" treatment with these compounds of General formula 1 (1.2.1 · HCl, 1.2.2 · HCl, 1.2.5 · HCl, 1.2.8 · HCl, 1.2.9 · HCl) and Tamiflu animals from the groups "treated with compounds" prevented their death completely.

The anti-influenza efficacy of the studied preparations of general formula 1 (1.2.1 · HCl, 1.2.2 · HCl, 1.2.5 · HCl, 1.2.8 · HCl, 1.2.9 · HCl) and Tamiflu is expressed in a decrease in the rate of weight loss in the treated mice groups compared to the viral control group. Animal weight loss is one of the clinical signs of the manifestation of influenza pneumonia. A greater decrease in animal weight indicates a more severe course of the disease. Mice were weighed at 1, 2, 3, 4, 5 days after infection, and then every other day until the 15th day of observation. It was found that in the viral control group, the animals lost the most weight 5 days after infection (about 10%). In contrast to viral control, in groups of animals treated with all of these compounds of the general formula 1 (1.2.1 · HCl, 1.2.2 · HCl, 1.2.5 · HCl, 1.2.8 · HCl, 1.2.9 · HCl) and Tamiflu , on average, no weight loss was observed. Starting from day 9, all animals in the groups "treated with compounds" actively and stably gained weight.

Thus, it has been shown that the treatment of influenza pneumonia in mice is highly effective with compounds of the general formula 1.

Claims (10)

1. (3R, 4R, 5S) -5-amino-4-acylamino-3- (1-ethyl-propoxy) cyclohex-1-ene-carboxylic acids, their esters of general formula 1 and their pharmaceutically acceptable salts and / or hydrates

where R1 is hydrogen, C ₁ -C ₇ alkyl, C ₂ -C ₇ alkenyl or C ₂ -C ₇ alkynyl optionally substituted with C ₃ -C ₆ cycloalkyl, phenyl, pyridyl, C ₁ -C ₃ alkoxy;
R2 is hydrogen; -NH ₂ ; C ₂ H ₅ O-; substituted C ₁ -C ₅ alkyl selected from —CH ₂ OH, —CH ₂ —NH ₂ , —CH = CH ₂ , —CH = CHCl, —C≡CH, —CH ₂ F, —CHF ₂ , —CF ₃ , -CH ₂ CF ₃ , cyclo-C ₃ H ₇ , --CF ₂ CF ₂ CF ₂ CF ₃ , -CF ₂ CF ₂ CF ₂ CHF ₂ , -CH ₂ CF ₂ CF ₂ CF ₂ CF ₃ . 2. The compounds according to claim 1, which are acids of general formula 1.1 and their esters of general formula 1.2

in which R1 represents C ₁ -C ₅ alkyl, C ₃ -C ₅ alkenyl or C ₃ -C ₅ alkynyl, and R2 has the above meaning. 3. The compounds according to claim 2, which are
(3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.1,
(3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.2,
(3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.3,
(3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.4,
(3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.5,
(3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.6,
(3R, 4R, 5S) -5-amino-4- (2,2-difluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.7,
(3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.8,
(3R, 4R, 5S) -5-amino-4- (3,3,3-trifluoro-propionylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.9,
(3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5-octafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1-en- carboxylic acid 1.1.10,
(3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,5-nonafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1- en-carboxylic acid 1.1.11,
(3R, 4S, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,6,6,6-undecafluorohexanoylamino) -3- (1-ethylpropoxy) - cyclohex-1-ene-carboxylic acid 1.1.12,
(3R, 4R, 5S) -5-amino-4- (cyclopropylcarbonyl-amino) -3- (1-ethylpropoxy) cyclohex-1-en-carboxylic acid 1.1.14,
(3R, 4R, 5S) -5-amino-4- (ethoxycarbonyl-amino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.15,
(3R, 4R, 5S) -5-amino-4-ureido-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.1.16,
(3R, 4R, 58) -4- (3-chloroprop-2-enoyl amino) -5-amino-3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.1.17.
ethyl ether (3R, 4R, 5S) -5-amino-4-formylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.1,
ethyl ether (3R, 4R, 5S) -5-amino-4- (2-hydroxy-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.2,
ethyl ester of (3R, 4R, 5S) -5-amino-4- (2-amino-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.3,
ethyl ester of (3R, 4R, 5S) -4-acryloylamino-5-amino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.4,
ethyl ester of (3R, 4R, 5S) -5-amino-4-propinoylamino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.5,
ethyl ether (3R, 4R, 5S) -5-amino-4- (2-fluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.6,
ethyl ester of (3R, 4R, 5S) -5-amino-4- (2,2-difluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.7,
ethyl ether (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.8,
ethyl ester of (3R, 4R, 5S) -5-amino-4- (3,3,3-trifluoro-propionylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.9,
ethyl ester (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5-octafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex-1- en-carboxylic acid 1.2.10,
ethyl ester (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,5-nonafluoro-pentanoylamino) -3- (1-ethylpropoxy) cyclohex- 1-en-carboxylic acid 1.2.11,
ethyl ester (3R, 4R, 5S) -5-amino-4- (2,2,3,3,4,4,5,5,6,6,6-undecafluorohexanoylamino) -3- (1-ethylpropoxy ) -cyclohex-1-ene-carboxylic acid 1.2.12,
ethyl ether (3R, 4R, 5S) -5-amino-4- (cyclopropylcarbonyl-amino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.14,
ethyl ester of (3R, 4R, 5S) -5-amino-4-ureido-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.15,
(3R, 4R, 5S) -5-amino-4- (ethoxycarboylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid ethyl ester 1.2.16,
ethyl ester of (3R, 4R, 5S) -4- (3-chloroprop-2-enoyl-amino) -5-amino-3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid 1.2.17,
allyl ether (3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene carboxylic acid 1.2.18,
(3R, 4R, 5S) -5-amino-4- (2,2,2-trifluoro-acetylamino) -3- (1-ethylpropoxy) cyclohex-1-ene-carboxylic acid prop-2-ynyl ester 1.2. 19. 4. The drug beginning with activity against neuraminidase, which is a compound according to any one of claims 1 to 3. 5. A pharmaceutical composition having antiviral activity and intended for the treatment of influenza and pneumonia due to influenza virus, comprising a drug onset according to claim 4 in a therapeutically effective amount. 6. The drug is in the form of tablets, capsules or injections, placed in a pharmaceutically acceptable package containing the pharmaceutical composition according to claim 5 in an effective amount. 7. The drug according to claim 6, for the treatment of pneumonia due to influenza virus. 8. A method of inhibiting neuraminidase activity, comprising the step of contacting it with a compound according to any one of claims 1 to 3. 9. The method according to claim 8, in which the neuraminidase is a flu virus neuraminidase. 10. A method for the prevention and treatment of influenza and pneumonia due to influenza virus, which consists in the introduction of a therapeutically effective amount of the drug substance according to claim 4, or a pharmaceutical composition according to claim 5, or a drug according to claim 6 or 7.