RU2712229C2 - Ethers of trihydroxyheptane acid as fpr2 receptor agonists - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: present invention relates to analogues of autocoids, in particular to derivatives of trihydroxyheptane acid ester of formula (I) (in which values of radicals are specified in the claim) and their use as medicinal agents. Present invention also relates to related subjects, including methods for preparing compounds, pharmaceutical compositions containing one or more compounds of formula (I), and in particular to use thereof as FPR2 receptor agonists.

EFFECT: ethers of trihydroxyheptane acid as agonists of FPR2 receptor are described.

5 cl, 1 dwg, 10 ex, 1 tbl

Description

The present invention relates to analogues of autocoids, in particular to derivatives of the trihydroxyheptaenoic acid ester of formula (I) and their use as medicaments. The present invention also relates to related matters, including processes for preparing compounds, pharmaceutical compositions containing one or more compounds of formula (I), and in particular, their use as FPR2 receptor agonists.

Autocoids are local biological signaling molecules, in particular, small lipid molecules synthesized from ω-3 polyunsaturated fatty acids: eicosapentaenoic (C 20: 5, EPA) and docosahexaenoic (C 22: 6, DHA). Compounds with this structure were first identified in 2000 during experimental inflammation in mice using lipidomics (liquid chromatography combined with tandem mass spectrometry) and biometrics (Serhan et al. Novel functional sets of lipid-derived mediators with antiinflammatory actions generated from omega-3 fatty acids via cyclooxygenase 2-nonsteroidal antiinflammatory drugs and transcellular processing // J. Exp. Medicine. - 2000. - T. 192. - No. 8. - C. 1197-1204).

Autocoids of the D-series (RD), which are polyhydroxylated derivatives of unsaturated fatty acids, were first found in exudates during the development of experimental inflammation in mice. The source for their synthesis is DHA. Endogenous DHA is a substrate for the enzyme 15-lipoxygenase, which converts DHA to 17S-hydroperoxy-DHA. This intermediate can then turn into several bioactive components, including RD1, 2, 3, and 4 (Hong et al. Novel docosatrienes and 17S-resolvins generated from docosahexaenoic acid in murine brain, human blood, and glial cells autacoids in anti-inflammation // J. Biol. Chem. - 2003. - T. 278. - No. 17. - C. 14677-14687).

RD1 is a powerful regulator of the activity of polymorphonuclear leukocytes. It inhibits the migration of neutrophils through the endothelium, blocks the transcription of IL-1β in microglial cells, and in experimental peritonitis in mice reduces the peritoneal infiltration by polymorphonuclear leukocytes (Serhan et al. Resolvins // J. Exp.Medicine. - 2002. - T. 196 . - No. 8. - C. 1025-1037; Hong et al, 2003). The introduction of RD1 before or after experimental renal ischemia reduces the morphological signs of damage (Duffield et al. Resolvin D series and protectin D1 mitigate acute kidney injury // J. Immunol. - 2006. - T. 177. - No. 9. - C. 5902 -5911). The use of certain polyhydroxylated derivatives of unsaturated fatty acids (Patent WO 2004014835 A2, published 02.19.2004; Patent US 8586073 B2, published 11.19.2013) and their derivatives for the treatment of pain (Patent CN 103193643 A, 07/10/2013), for scar healing (Patent US 9463177 B2, published 11/10/2016), etc.

An inflammatory response plays a central role in protecting the body against invading pathogens and tissue damage. The initiation of this response is an active process, and recent studies have shown that resolving inflammation, as well as its initiation, is an active process mediating a number of endogenous lipid derived molecules that can “turn off” the inflammatory cascade and promote homeostasis (Serhan, Savill Resolution of inflammation: the beginning programs the end // Nature immunology. - 2005. - T. 6. - No. 12. - C. 1191-1197). These lipids are mainly derived from the enzymatic reaction of endogenous lipoxygenase and cyclooxygenase with C-20 and C-22 LC-PUFA (long chain polyunsaturated fatty acids) to form oxidized products or oxylipins in the area of inflammation. When administered to animals, autocoids have strong anti-inflammatory activity (Gilroy et al. Inflammatory resolution: new opportunities for drug discovery // Nature reviews. Drug discovery. - 2004. -T. 3. - No. 5. - C. 401-416; Serhan , Savill, 2005; Hong et al, 2003; Serhan et al, 2002).

Recently, the search for new compounds based on polyhydroxylated derivatives of unsaturated fatty acids has focused on obtaining stable analogues with an improved pharmacological profile (Guilford et al, 2004; Dangi B. et al. Metabolism and biological production of resolvins derived from docosapentaenoic acid (DPAn-6) // Biochem. Pharmacol - 2010. - T. 79. - No. 2. - C. 251-260).

Among the many publications on the effects of RD1 (Serhan, Pro-resolving lipid mediators are leading for resolution physiology. // Nature. - 2014. - T. 510.- C. 92-101), RD1 decreases TNF-alpha signaling ( Lee et al. Resolvin D1 stimulates efferocytosis through p50 / p50-mediated suppression of tumor necrosis factor-alpha expression. // J Cell Sci. - 2013 .-- T. 126. - C. 4037-4047), reduces pain (Ji et al. Emerging targets in neuroinflammation-driven chronic pain. // Nat Rev Drug Discov. - 2014.- T. 13 - C. 533-548), plays an important role in bone disorder (McCauley et al. Cutting edge: Parathyroid hormone facilitates macrophage efferocytosis in bone marrow via proresolving mediators resolvin D1 and resolvin D2. // J Immunol. - 2014. - T. 193. - S.26-29), performs a protective function in the experiment lnyh models of ischemia-reperfusion (Martin et al. Resolvin D1 and lipoxin A4 improve alveolarization and normalize septal wall thickness in a neonatal murine model of hyperoxia-induced lung injury. // PLoS One. - 2014. - T. 9. - C e98773), fibromyalgia (Klein et al. Effects of D-series resolvins on behavioral and neurochemical changes in a fibromyalgia-like model in mice. // Neuropharmacology. - 2014.- T. 86C . - C. 57-66), colitis (Bento et al. Omega-3 fatty acid-derived mediators 17 (R) -hydroxy docosahexaenoic acid, aspirin-triggered resolvin D1 and resolvin D2 prevent experimental colitis in mice.// J Immunol . - 2011. - 187: 1957-1969), has an antiallergic effect (Nelson et al. ALX / FPR2 receptor for RvD1 is expressed and functional in salivary glands. // Am J Physiol Cell Physiol. - 2014.- T. 306. - C 178-C185). Taking into account the protective effect of RD1, its extremely high susceptibility to oxidation and high cost, we proposed a new analogue of RD1 with enhanced chemical stability and ease of general organic synthesis, while maintaining biological activity.

The biological effects of all eicosanoids, including autocoids, are mediated by specific eicosanoid receptors. Most eicosanoid receptors transmit a signal through G proteins. Typically, the effects of autocoids are mediated by CMKLR1 receptors (chemokine-like receptor 1, also referred to as ChemR23), GPR32 (G-protein-bound receptor 32), LTB4R (leukotriene B4 receptor) and LXA4R (lipoxin receptor, or FPR2 / ALXR receptor) .

FPR2 - formyl peptide receptor 2 - formulated a chemotaxis-regulating peptide receptor that belongs to the class of G-protein linked receptors. The lipid metabolite, lipoxin A4 (LXA4) and its analogues have been found to bind to the FPR2 receptor with high affinity and enhance the production of arachidonic acid (Chiang et al. The lipoxin receptor ALX: potent ligand-specific and stereoselective actions in vivo // Pharmacol. reviews. - 2006. - T. 58. - No. 3. - C 463-487). LXA4 has high anti-inflammatory activity. On models of skin inflammation, peritonitis, colitis, mesangioproliferative nephritis, pleurisy, asthma, cystic fibrosis, sepsis, periodontitis, etc. (Schwab, Serhan Lipoxins and new lipid mediators in the resolution of inflammation // Cur. Opinion pharmacol. - 2006. - T 6. - No. 4. - C. 414-420).

Biological characteristics of FPR2 agonists include, but are not limited to, migration / activation of monocytes / macrophages / microglial cells / dendritic cells, migration / activation of neutrophils, regulation of activation, proliferation and differentiation of lymphocytes, regulation of inflammation, regulation of production and / or release of cytokines, regulation production and / or release of pro-inflammatory mediators, regulation of the immune response.

The present invention relates to analogues of autocoids - esters of trihydroxyheptaenoic acid, which are non-peptide agonists of the FPR2 receptor. The compounds are useful in the prevention or treatment of diseases that respond to the modulation of the FPR2 receptor, such as inflammatory diseases (preferably metabolic syndrome and type 2 diabetes), obstructive airways diseases, allergic conditions, HIV-mediated retroviral infections, cardiovascular disorders, neuroinflammation neurological disorders, prion-mediated pain and amyloid-mediated disorders (preferably Alzheimer's disease EPA); in addition, they are useful for the prevention or treatment of autoimmune diseases and for modulating immune responses.

Various embodiments of the present invention are described below.

1) The present invention relates to esters of trihydroxyheptaenoic acid of the formula (I),

wherein,

R is phenyl, (C ₂ -C ₄ ) alkyl, benzyl, methoxybenzyl or phenethyl; and salts (preferably pharmaceutically acceptable salts) of such compounds.

Compounds of formula (I) according to embodiment (1) may contain one or more stereogenic or asymmetric centers, such as one or more asymmetric carbon atoms. Substituents at the double bond may be in the (Z) - or (E) -configuration, unless otherwise indicated. Thus, the compounds of formula (I) can be in the form of mixtures of stereoisomers, or preferably in the form of pure stereoisomers. Mixtures of stereoisomers can be separated according to methods known to a person skilled in the art.

In the following paragraphs, definitions of various chemical fragments of the compounds of the present invention are given, and they are applicable throughout the specification and claims, unless other definitions provide broader or narrower definitions.

The term “alkyl”, when used alone or in combination, means a linear or branched chain alkyl group containing from 2 to 4 carbon atoms. Typical examples of alkyl groups include ethyl, isopropyl and tert-butyl.

2). Preferred compounds of formula (I) as defined in embodiment (1) are selected from the group consisting of:

(5R, 6S, Z) -Ethyl-5,6,7-trihydroxyhept-2-enoate

(5R, 6S, E) -Ethyl-5,6,7-trihydroxyhept-2-enoate

(5R, 6S, E) -tert-Butyl-5,6,7-trihydroxyhept-2-enoate

(5S, 6R, E) -Benzyl-5,6,7-trihydroxyhept-2-enoate

(5R, 6S, E) -Benzyl-5,6,7-trihydroxyhept-2-enoate

(5S, 6R, E) -Phenyl-5,6,7-trihydroxyhept-2-enoate

(5R, 6S, E) -Phenyl-5,6,7-trihydroxyhept-2-enoate

(5R, 6S, E) - (4-Methoxybenzyl) -5,6,7-trihydroxyhept-2-enoate

(5R, 6S, E) -Phenethyl-5,6,7-trihydroxyhept-2-enoate

The term "pharmaceutically acceptable salts" means non-toxic addition salts with an inorganic or organic acid and / or base (see, for example, Gould PL Salt selection for basic drugs // International Journal of Pharmaceutics. - 1986. - T. 33. - No. 1 -3. - C. 201-217).

The compounds of formula (I) corresponding to any of embodiments (1) to (2), or their pharmaceutically acceptable salts, are suitable for use as medicaments. In particular, the compounds of formula (I) modulate the FPR2 receptor, i.e. they act as agonists of the FPR2 receptor, and are useful in the prevention or treatment of diseases that respond to the activation of the FPR2 receptor.

In particular, the compounds of formula (I) corresponding to any of embodiments (1) to (2), or pharmaceutically acceptable salts thereof, are suitable for the prevention or treatment of diseases selected from the group consisting of inflammatory diseases, preferably metabolic syndrome, diabetes mellitus 2 type, rheumatoid arthritis, acute lung damage, asthma, inflammatory bowel disease and Alzheimer's disease.

The present invention also relates to the use of a compound of formula (I) corresponding to any of embodiments (1) to (2) for the preparation of pharmaceutical compositions for the treatment and / or prophylaxis of the above diseases.

The present invention also relates to pharmaceutically acceptable salts and to pharmaceutical compositions and preparations of compounds of formula (I) corresponding to any of embodiments (1) to (2).

The pharmaceutical composition of the present invention contains at least one compound of formula (I) corresponding to any of embodiments (1) to (2) (or a pharmaceutically acceptable salt thereof) as an active agent and optionally carriers and / or diluents, and / or excipients.

The compounds of formula (I) corresponding to any of embodiments (1) to (2) and their pharmaceutically acceptable salts in accordance with the present invention are preferably intended for oral administration to a human or animal. The pharmaceutical composition may also be intended to be administered by any other method in which the active ingredients can be effectively absorbed and used, for example, intravenously, subcutaneously, intramuscularly, intranasally, rectally, vaginally or topically.

In a specific embodiment of the present invention, the pharmaceutical composition is formed in the form of a capsule, which may also be a microcapsule, a powder generating, or sachet. The capsule may flavor. This embodiment also includes a capsule where both the capsule and the encapsulated composition of the present invention are flavored. Through aromatization, the capsule becomes more attractive to the consumer. For the therapeutic applications discussed above, the dosage administered will, of course, vary with the compound used, with the route of administration, the treatment desired and the disorder indicated.

The pharmaceutical composition may be prepared in a daily dose, for example, from 1 μg to 10 mg of the compound. Formulas (I) Daily dosing refers to dosing within 24 hours.

The preparation of pharmaceutical compositions can be carried out according to methods that should be known to a person skilled in the art (see, for example, Remington, The Science and Practice of Pharmacy, 21st Edition (2005), Part 5, "Pharmaceutical Manufacturing" [published by Lippincott Williams & Wilkins]), by introducing the described compounds of formula (I) or their pharmaceutically acceptable salts, optionally in combination with other therapeutically useful substances, in an administered dosage form together with suitable non-toxic inert therapeutically compatible solid or liquid carriers and when necessary with conventional pharmaceutical excipients.

The present invention also relates to a method for preventing or treating a disease or disorder specified in the present invention, comprising administering to a subject a compound of formula (I) corresponding to any one of embodiments (1) to (2), or a pharmaceutically acceptable salt thereof in a pharmaceutically active amount.

Any reference to a compound of formula (I) in this text should be understood as referring to salts (and preferably pharmaceutically acceptable salts) of such compounds, if this is suitable or expedient. Of course, the preferences indicated for the compounds of formula (I), with corresponding changes, apply to the salts and pharmaceutically acceptable salts of the compounds of formula (I). This also applies to these compounds as medicaments, to pharmaceutical compositions containing these compounds as active active agents, or to the use of these compounds for the preparation of a medicament for the treatment of diseases of the invention.

The compounds of formula (I) can be obtained by the methods described below, by the methods described in the examples or by similar methods. The optimal reaction conditions may vary depending on the particular reagents or solvents used, but such conditions can be determined by a person skilled in the art using standard optimization procedures.

Unless otherwise indicated, representative R groups are as defined for formula (I). Other abbreviations used are defined in the experimental section.

Pharmacological properties and a method for producing the trihydroxyheptaenoic acid esters of the formula (I) of the present invention are not described in the literature.

A. Synthesis of end products

The present invention will now be described in more detail using the following examples, which should not be construed as limiting the present invention. Compounds of formula (I) can be prepared from DOR by reaction with the corresponding triphenylphosphoranylideneacetate derivative at the boiling point in a suitable solvent.

Example 1

Compounds of structure 1 or 2 were obtained by the reaction of a solution of DOR (1.00 g, 7.46 mmol) and ethyl 2- (triphenylphosphoranilidene) acetate (2.86 g, 8.2 mmol) in 15 ml of dry THF under reflux for 2 h, after which the reaction mixture was concentrated , the residue was purified by column chromatography on silica gel (eluent CH ₂ Cl ₂ : MeOH, 97: 3). The obtained cis and trans isomers were purified by column chromatography repeatedly and recrystallized from a mixture of MTBE: pentane, 1: 6. The colorless precipitates formed at -15 ° C were filtered off and washed with pentane. Received 15 mg (1%) of compound of structure 1 with a purity (HPLC) of 100%, mass spectrum: calculated for C ₉ H ₁₆ NaO ₅ (M + Na ⁺ ): 227.0895, found 227.0893 and 220 mg (14%) of compound of structure 2 purity (HPLC) 97.6%, mass spectrum: calculated for C9H16NaO5 (M + Na ⁺ ) 227.0895, found 227.0889. Both compounds are obtained in the form of white powders.

Example 2

The compound of structure 3 was obtained by the reaction of a solution of DOR (0.95 g, 7.1 mmol) and tert-butyl 2- (triphenylphosphoranilidene) acetate (2.93 g, 7.8 mmol) in 15 ml of dry THF at boiling point and holding for 2 h, after which the reaction mixture was concentrated, the residue was purified by silica gel column chromatography (eluent CH ₂ Cl ₂ : EtOAc: MeOH, 65: 33: 2). The resulting trans isomer was purified by column chromatography repeatedly and recrystallized from a mixture of MTBE: pentane, 1: 4. The colorless precipitates formed at -15 ° C were filtered off and washed with pentane. Received 300 mg (18%) of compound of structure 3 as a colorless fusible solid with a purity (HPLC) of 100%, mass spectrum: calculated for C ₁₁ H ₂₀ NaO ₅ (M + Na ⁺ ): 255.1203, found 255.1208.

Example 3

The compound of structure 4 can be obtained by the reaction of a solution of DOR (1.34 g, 10 mmol) and benzyl 2- (triphenylphosphoranilidene) acetate (6.16 g, 15 mmol) in 35 ml of dry THF under reflux for 2 hours, after which the reaction mixture was concentrated. the residue was purified by silica gel column chromatography (eluent, CH ₂ Cl ₂ : MeOH, 94: 6). The resulting trans isomer was recrystallized from MTBE. The colorless precipitate formed at 5 ° С was filtered off and washed with chilled MTBE. Received 420 mg (16%) of compound of structure 4 in the form of a crystalline white powder with a purity (HPLC) of 99.6%, mass spectrum: calculated for C ₁₄ H ₁₈ NaO ₅ (M + Na ⁺ ): 289.1046, found 289.1044.

Example 4

The compound of structure 5 can be obtained by the reaction of a solution of DOR (1.50 g, 11.2 mmol) and benzyl 2- (triphenylphosphoranilidene) acetate (6.90 g, 16.8 mmol) in 40 ml of dry THF by boiling for 2 h, after which the reaction mixture was concentrated, the residue purified by KX (eluent CH ₂ Cl ₂ : MeOH, 97: 3). The resulting isomers were recrystallized from MTBE. Precipitations at 5 ° C were filtered off and washed with chilled MTBE. Received 615 mg (20%) of compound of structure 5 in the form of a white powder with a purity (HPLC) of 99.9%, mass spectrum: calculated for C ₁₄ H ₁₈ NaO ₅ (M + Na ⁺ ): 289.1046, found 289.1040.

Example 5

Obtaining an intermediate product 1 (phenyl-2-iodoacetate). 13.49 g (90 mM) of sodium iodide and 14.5 g (85 mM) of phenyl-2-chloroacetate were dissolved in 54 ml of CH ₃ COCH ₃ , the mixture was stirred for 3 hours. The precipitate was filtered off and washed with CH ₃ COCH ₃ . The solution was evaporated, 30 ml of methylene chloride was added to the residue, the mixture was filtered through FS. The solvent was evaporated in vacuo, the reaction product was recrystallized from ethanol. The yield of intermediate 1 was 18.16 g (82%).

Preparation of Intermediate 2 (phenyl-2- (triphenylphosphoranylidene) acetate). 18.16 g (69.3 mmol) of triphenylphosphine was dissolved in 200 ml of C ₆ H ₆ , the mixture was filtered through FS. 18.16 g (69.3 mmol) of intermediate 1 was added to the solution. After 16 hours, 100 ml of PE was added to the mixture, the mixture was cooled to 3-5 ° C, the precipitate was filtered off, washed with PE and dried. The dried precipitate was added to a mixture of 100 ml of a 0.5 M NaOH solution with 200 ml of CH ₂ Cl ₂ cooled to 0-5 ° C. The organic layer was separated, the aqueous layer was washed with 20 ml of CH ₂ Cl ₂ . The combined organic extracts were dried with Na ₂ SO ₄ , the solvent was distilled off. The yield of intermediate 2 as a yellowish crystallizing gummy substance was 18.41 g (67%).

The compound of structure 6 can be obtained by the reaction of a solution of DOR (1.0 g, 7.5 mmol) and intermediate 2 (3.96 g, 10.0 mmol) in 25 ml of dry THF during boiling, after which the reaction mixture was concentrated, the residue was purified by KC (eluent CH ₂ Cl ₂ : MeOH, 97: 3). The resulting trans isomer was recrystallized from MTBE. The colorless precipitate formed at 5 ° С was filtered off and washed with chilled MTBE. Received 50 mg (26%) of compound of structure 6 in the form of a white powder with a purity (HPLC) of 98.5%, mass spectrum: calculated for C ₁₃ H ₁₆ NaO ₅ (M + Na ⁺ ): 275.0895, found 275.0897.

Example 6

Obtaining an intermediate product 1 (see for structure 6).

Preparation of Intermediate 2 (phenyl-2- (triphenylphosphoranylidene) acetate). 18.16 g (69.3 mmol) of triphenylphosphine was dissolved in 200 ml of C ₆ H ₆ , the mixture was filtered through FS. 18.16 g (69.3 mmol) of intermediate 1 was added to the solution. After 16 h, 100 ml of petroleum ether was added to the mixture, the mixture was cooled to 3-5 ° C, the precipitate formed was filtered off, washed with PE and dried. The dried precipitate was added to a mixture of 100 ml of a 0.5 M NaOH solution with 200 ml of CH ₂ Cl ₂ cooled to 0-5 ° C. The organic layer was separated, the aqueous layer was washed with 20 ml of CH ₂ Cl ₂ . The combined organic extracts were dried with Na ₂ SO ₄ , the solvent was distilled off. The yield of intermediate 2 as a yellowish crystallizing gummy substance was 18.41 g (67%).

The compound of structure 7 was obtained by the reaction of a solution of DOR (1.50 g, 11.2 mmol) and intermediate 2 (6.66 g, 16.8 mmol) in 40 ml of dry THF during boiling, after which the reaction mixture was concentrated, the residue was purified by KC (eluent CH ₂ Cl ₂ : MeOH, 97: 3). The resulting trans isomer was recrystallized from MTBE. The colorless precipitate formed at 5 ° С was filtered off and washed with chilled MTBE. Received 315 mg (20%) of compound of structure 7 in the form of a white powder with a purity (HPLC) of 96.6%, mass spectrum: calculated for C ₁₃ H ₁₆ NaO ₅ (M + Na ⁺ ): 275.0895, found 275.0885.

Example 7

Preparation of Intermediate 1 ((4-methoxybenzyl) -2-chloroacetate). 10.9 ml (12.0 g, 87 mmol) of anise alcohol and 7 ml (6.9 g, 87 mmol) of abs. pyridine was dissolved in 80 ml of methylene chloride, after which the mixture was cooled in an ice bath. Upon cooling, 6.9 ml (9.8 g, 87 mmol) of chloroacetyl chloride were added dropwise to the mixture, and kept until a CT was reached. After 16 hours, the precipitate was filtered off and washed with CH ₂ Cl ₂ . The solution was washed with a 1% HCl solution and dried with anhydrous Na ₂ SO ₄ . The solvent was evaporated in vacuo. The yield of intermediate 1 was 17.91 g (91%) as a colorless oily substance.

Preparation of Intermediate 2 ((4-Methoxybenzyl) -2-iodoacetate). 3.75 g (25 mmol) of sodium iodide and 4.29 g (20 mmol) of intermediate 1 were dissolved in 15 ml of CH ₃ COCH ₃ , the mixture was stirred for 3 hours. The precipitate was filtered off and washed with CH ₃ COCH ₃ . The solution was evaporated, 30 ml of CH ₂ Cl _{2 was} added to the residue, the mixture was filtered through FS. The solvent was evaporated in vacuo. Yield of intermediate 2 5.20 g (85%), yellowish oily substance.

Preparation of Intermediate 3 ((4-Methoxybenzyl) -2- (triphenylphosphoranilidene) acetate). 4.20 g (16 mmol) of triphenylphosphine was dissolved in 200 ml of C6H6, and the mixture was filtered through FS. 4.81 g (16 mmol) of intermediate 2 was added to the solution. After 16 h, 100 ml of PE was added to the mixture, the mixture was cooled to 3-5 ° C, the precipitate formed was filtered off, washed with petroleum ether and dried. The dried precipitate was added to a mixture of 100 ml of a 0.5 M NaOH solution with 200 ml of CH ₂ Cl ₂ cooled to 0-5 ° C. The organic layer was separated, the aqueous layer was washed with 20 ml of CH ₂ Cl ₂ . The combined organic extracts were dried with Na ₂ SO ₄ , the solvent was distilled off. The yield of intermediate 3 as a yellowish crystallizing gummy substance was 6.61 g (94%).

The compound of structure 8 was obtained by the reaction of a solution of DOR (1.34 g, 10 mmol) and intermediate 3 (6.61 g, 15 mmol) in 25 ml of dry THF under reflux for 2 h, after which the reaction mixture was concentrated, the residue was purified by KC (eluent CH ₂ Cl ₂ : MeOH, 97: 3). The resulting trans isomer was recrystallized twice from MTBE. The colorless precipitate formed at 5 ° С was filtered off and washed with chilled MTBE. Received 296 mg (10%) of compound of structure 8 in the form of a white powder with a purity (HPLC) of 99.4%, mass spectrum: calculated for C ₁₅ H ₂₁ O ₆ (M + H ⁺ ): 297.32, found 297.30.

Example 8

Obtaining an intermediate product 1 (phenethyl-2-iodoacetate). 3.75 g (25 mmol) of sodium iodide and 3.97 g (20 mmol) of phenethyl-2-chloroacetate were dissolved in 15 ml of CH ₃ COCH ₃ , the mixture was stirred for 3 hours. The precipitate was filtered off and washed with CH ₃ COSH ₃ . The solution was evaporated, 30 ml of CH ₂ Cl _{2 was} added to the residue, the mixture was filtered through FS. The solvent was evaporated in vacuo. The yield of intermediate 1 was 4.81 g (83%) as a yellow oily substance.

Preparation of Intermediate 2 (phenethyl-2- (triphenylphosphoranilidene) acetate). 4.35 g (16.6 mmol) of triphenylphosphine was dissolved in 200 ml of C6H ₆ , and the mixture was filtered through FS. 4.81 g (16.6 mmol) of intermediate 1 was added to the solution. After 16 h, 100 ml of PE was added to the mixture, the mixture was cooled to 3-5 ° C, the precipitate formed was filtered off, washed with PE and dried. The dried precipitate was added to a mixture of 100 ml of a 0.5 M NaOH solution with 200 ml of CH ₂ Cl ₂ cooled to 0-5 ° C. The organic layer was separated, the aqueous layer was washed with 20 ml of CH ₂ Cl ₂ . The combined organic extracts were dried with Na ₂ SO ₄ , the solvent was distilled off. The yield of intermediate 2 was 6.9 g (98%) as a yellowish crystallizing gummy substance. The compound of structure 9 was obtained by the reaction of a solution of DOR (1.43 g, 10.7 mmol) and intermediate 2 (6.9 g, 16.3 mmol) in 20 ml of dry THF during boiling for 2 h, after which the reaction mixture was concentrated, the residue was purified by KC (eluent CH ₂ C1 ₂ : MeOH, 97: 3). The resulting trans isomer was recrystallized from MTBE. The colorless precipitate formed at 5 ° С was filtered off and washed with chilled MTBE. Received 325 mg (11%) of compound of structure 9 in the form of a white powder with a purity (HPLC) of 99.0%, mass spectrum: calculated for C ₁₅ H ₂₁ O ₅ (M + H ⁺ ): 281.14, found 282.20.

I. Chemical data

General Provisions All temperatures are in degrees Celsius (° C). Unless otherwise indicated, reactions occur during CT.

Flash column chromatography (PF) was performed using 60 Merck silica gel (0.063-0.200 mm) or Macherey-Nagel silica gel (0.063-0.200 mm): CH ₂ Cl ₂ : MeOH in the ratio 97: 3 was used for elution.

LC-MS conditions (unless otherwise indicated): Analytical: pump for supplying two components Dionex HPG-3000, MS: Thermo MSQ, DDM: Dionex PDA 3000, IDSR: PolymerLab ELS 2100. Column: Ascentis Express C18 2.7 μm , 2.1 × 30 mm ID (internal diameter), manufactured by Sigma-Aldrich, thermostatically controlled in a Dionex chamber TCC-3200. Eluents: A: H ₂ O + 0.05% NH ₄ OH + 2% AcCN; B: AcCN. Method: Gradient mode: 5% V → 95% V in 2.00 min. Flow rate: 1.8 ml / min. Detection: UV / view, t _R are given in minutes.

HPLC conditions (unless otherwise indicated): Column: Luna C18 (2) 5 μm, 4.6 × 150 mm, manufactured by Phenomenex. Eluent: 0.03% TFA + AcCN 88:12. Method: Gradient mode: 5% V → 95% V in 2.00 min. Flow rate: 1.0 ml / min. Detection: UV / view, t _R are given in minutes.

NMR: Bruker Avance 400 (400 MHz); Varian Mercury 300 (300 MHz); chemical shifts are given in ppm relative to the solvent used; multiplicity: s = singlet, d = doublet, t = triplet, q = quadruplet, p = pentiplet, hex = sextet, hept = heptet, m = multiplet, br = wide, spin-spin coupling constants are given in hertz.

II. Biological research

DESCRIPTION OF FIGURES

In FIG. 1 shows the dependence of the inhibitory effect on the concentration of a compound of structure 2 on the COX-2 enzyme

On the abscissa, the concentration of compound of structure 2 in μg / ml; along the ordinate axis, inhibition of COX 2 enzyme activity in% of control.

In vitro study

Example 9. The agonistic activity of the compounds of formula (I) with respect to the FPR2 receptor was determined by the experimental procedure described below.

The synthesis of pro-inflammatory cytokines is initiated by intracellular signaling, one of the links of which are p38, JNK1 / 2 and ERK1 / 2 MAP kinases. When cells are stimulated with inflammatory factors (bacterial agents, cytokines, viruses, etc.), MAP kinases are phosphorylated and activate factors that trigger the transcription of inflammatory mediators (TNFa, IL-1, IL-6, IL-8, COX-2, arachidonic acid and et al.) (Yang et al, 2003; Dumitru et al, 2000; Kyriakis et al, 2001).

In accordance with this, it was suggested that a decrease in the phosphorylation of MAP kinases, in particular, p38 kinase, can provide an anti-inflammatory effect of the tested substances by reducing the production of pro-inflammatory cytokines.

Experimental procedure: Determination of the effect of compounds of formula (I) on the LPS-induced phosphorylation of p38 MAP kinase in a transplanted culture of human monocytes of the U937 line.

The cell culture was incubated with compounds of formula (I) at various concentrations for 1 h at 37 ° C and 5% CO ₂ , then lipopolysaccharide (LPS) of the E. coli bacteria cell wall was added and kept for 1 h under the same conditions. After this period, the content of phosphorylated forms and the total amount (phosphorylated and non-phosphorylated forms) of p38 MAP kinase were determined in the culture in order to determine the level of activation (phosphorylation) of p38 MAP kinase. The method of determination is immunoblot. The study was conducted in comparison with a specific inhibitor of MAP kinase activation p38 SB203580 (5 μM (1.88 μg / ml))

The agonistic activity of the exemplified compounds with respect to the FPR2 receptor (amount of phospho-MAPK,% of control) is presented in table 1.

Estimation of the amount of phosphorylated forms of p38 MAP kinase in the implemented model showed that structures 2, 3 and 9 have the most pronounced effect on p38 MAP kinase activation. These compounds have an inhibitory effect on p38 MAP kinase activation in a wide range of concentrations. Substances of structures 5 and 6 influenced the activation of p38 MAP kinase in the concentration range from 4 μg / ml to 1 μg / ml.

Example 10. The activity of the compounds of formula (I) with respect to the enzymes cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) was determined by the experimental procedure described below.

The development of inflammation is ensured by the synthesis of a complex of pro-inflammatory mediators that stimulate most of the further processes in the development of the inflammatory response and activation of various types of cells involved in the maintenance and regulation of inflammation, including all types of leukocytes, dendritic cells, T and B lymphocytes, endothelial and epithelial cells, fibroblasts et al. (Holgate S.T. et al. Roles of cysteinyl leukotrienes in airway inflammation, smooth muscle function, and remodeling // J. Allergy Clin Immunol. 2003. Vol. 111. No. 1. P. 18-34.)

One of the possible mechanisms of the anti-inflammatory effect of various drugs can be the inhibition of the arachidonic acid cascade, the synthesis of prostaglandins and leukotrienes (Holgate S.T. et al., 2003). One of the key enzymes of this cascade of cyclooxygenase 1/2 (COX1 / 2) are promising targets for the search for new non-steroidal anti-inflammatory drugs (NSAIDs) (Kumar K.A. High-through screening assays for cyclooxygenase-2 and 5-lipoxygenase, the targets for inflammatory disorders // Ind. J. Biochem. Biophysics. 2011. Vol. 48. P. 256-261).

The effect of the synthesized compounds on the enzymatic activity of COX-1/2 was evaluated using a commercially available test system COX (human) inhibitory screening assay kit (Cayman Chemicals, USA).

Some of the compounds covered by the present invention are effective COX 1/2 enzyme inhibitors (compounds 2, 5, and 9). Moreover, the compound of structure 2 is a selective inhibitor of COX 2 and its inhibitory effect is similar to indomethacin, which is a non-selective inhibitor, which can be a mechanism of anti-inflammatory action of drugs based on it (see Fig. 1). The IC50 for the compound of structure 2 was about 0.21 ± 0.01 μg / ml (1.0 ± 0.03 μM), which is comparable to the non-specific COX inhibitor indomethacin: IC50 = 0.69 μM.

Claims (15)

1. The compound of formula (I),

in which R is benzyl, methoxybenzyl, phenyl or phenethyl; or (5R, 6S, E) -tert-Butyl-5,6,7-trihydroxyhept-2-enoate. 2. The compound according to claim 1, selected from the group including: (5R, 6S, E) -tert-Butyl-5,6,7-trihydroxyhept-2-enoate (5S, 6R, E) -Benzyl-5,6,7-trihydroxyhept-2-enoate (5R, 6S, E) -Benzyl-5,6,7-trihydroxyhept-2-enoate (5S, 6R, E) -Phenyl-5,6,7-trihydroxyhept-2-enoate (5R, 6S, E) -Phenyl-5,6,7-trihydroxyhept-2-enoate (5R, 6S, E) - (4-Methoxybenzyl) -5,6,7-trihydroxyhept-2-enoate (5R, 6S, E) -Phenethyl-5,6,7-trihydroxyhept-2-enoate. 3. The compound of claim 1 for use as a medicament for the prevention or treatment of a disease that responds to activation of the FPR2 receptor. 4. A pharmaceutical composition having agonistic activity against the FPR2 receptor and containing, as an active substance, a compound according to any one of claims. 1, 2 in pharmaceutically active amount and at least one therapeutically inert excipient. 5. The use of compounds according to any one of paragraphs. 1, 2 or (5R, 6S, E) -ethyl-5,6,7-trihydroxyhept-2-enoate for the preparation of a medicament for the prevention or treatment of a disease responding to modulation of the FPR2 receptor selected from metabolic syndrome, diabetes mellitus 2 types, rheumatoid arthritis, acute lung damage, asthma, inflammatory bowel disease and Alzheimer's disease.

Images