RU2613633C1 - Chloride 4-[(1e)-1-(6-chloro-4-oxo-4n-chromen-3-yl)-4-methylpent-1-en-3-yl]morpholine-4-yl, method of producing thereof and its antituberculosis effect - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to organic chemistry, namely to chloride 4-[(1E)-1-(6-chloro-4-oxo-4H-chromen-3-yl)-4-methylpent-1-en-3-yl]morpholine-4-yl of formula I

and method of producing thereof.

EFFECT: new heterocyclic compound, may be used as a potential antituberculosis drug.

3 cl, 5 dwg, 2 tbl, 2 ex

Description

The group of inventions relates to the field of organic and medical chemistry, namely to a new individual multifunctional heterocyclic compound - chloride 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent -1-en-3-yl] morpholin-4-th formula I, the method for its preparation and biological activity, which can be used in organic synthesis and in medicine as a potential anti-tuberculosis drug

The scientific and technical literature describes several methods for producing 4-allyl morpholines containing aromatic substituents. Thus, allyl morpholines (IV) were obtained by the interaction of phenylallenes (II) with morpholine (III) [Nishina, N. Gold-catalyzed hydrofunctionalization of allenes with nitrogen and oxygen nucleophiles and its mechanistic insight / N. Nishina, Y. Yamamoto // Tetrahedron. - 2009. - V. 65. - P. 1799-1808]. The reaction was carried out by heating in toluene under catalysis with a gold complex and silver triflate:

A similar reaction for the preparation of allyl morpholine (VI) was the addition of morpholine (III) at the terminal double bond of the diene (V) [Goldfogel, M.J. Intermolecular hydroamination of 1,3-dienes catalyzed by bis (phosphine) carbodicarbene-rhodium complexes / M.J. Goldfogel, C.C. Roberts, S.J. Meek // Journal of American Chemical Society. - 2014. - V. 136. - P. 6227-6230]. Catalysis according to this method was carried out by a complex of rhodium and silver tetrafluoroborate by heating the reagents in a chlorobenzene medium:

A literature also discloses a method for producing phenyl substituted allyl morpholines (VIII) with catalysis by a complex of palladium and triphenylphosphine [Singh, K. Facile synthesis of Z-alkenes via uphill catalysis / K. Singh, S.J. Staig, J.D. Weaver // Journal of American Chemical Society. - 2014. - V. 136. - P. 5275-5278]. The reaction between ether (VII) and morpholine (III) was carried out in dichloromethane:

As discovered by the authors of the work [Evans, R.W. A simple catalytic mechanism for the direct coupling of α-carbonyls with functionalized amines: a one-step synthesis of plavix / R.W. Evans, J.R. Zbieg, S. Zhu, W. Li, D.W.C. MacMillan // Journal of American Chemical Society. - 2013. - V. 135. - P. 16074-16077], heating a mixture of compound (IX) with aliphatic aldehyde (X) and morpholine (III) in the presence of divalent copper bromide also leads to phenylallyl morpholines (XI)

At sufficiently high yields (46-95%) of the products, the above methods for the preparation of allyl morpholines also have drawbacks that complicate the further industrial application of these syntheses. These include:

- The use of complex and inaccessible complexes of gold and rhodium, as well as silver and copper salts, as catalysts. The presence of heavy metals will complicate the further purification of the reaction products for use as medicines.

- All of these processes are carried out at elevated temperatures. Considering that morpholine (III) used in the syntheses is easily oxidized when heated by atmospheric oxygen, an inert atmosphere (for example, nitrogen) must be created for the reaction.

- These synthesis methods are designed to obtain groups of substances having an aromatic substituent.

From the patent and scientific and technical literature, neither a method of obtaining a new compound claimed by the authors, allyl morpholine with a chromene substituent (I), nor its structure itself have been identified.

The objective of the proposed group of inventions is the creation of a new compound not described in the literature - chloride 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent-1-en-3- il] morpholin-4-Ia (I), which will expand the range of potential anti-TB drugs.

The technical results to which the group of inventions is directed are to obtain a new multifunctional compound (I), which could potentially be used in medicine as a medicine; development of a simple method for its synthesis; identified anti-tuberculosis activity of this compound.

The problem is solved by the fact that a new compound is synthesized - chloride 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholine -4-th formula I

The problem is also solved by the fact that the compound has anti-tuberculosis activity.

The problem is also solved by the fact that the method of producing chloride 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholin- The 4th formula I is carried out by reacting 3- (4-oxo-6-chloro-4H-chromen-3-yl) acrylic acid with 4- (2-methylprop-1-en-1-yl) morpholine in the ratio 1: 1.5 in dichloroethane at a temperature of 18-20 ° C, followed by isolation of the target product in the form of a precipitate upon exposure to the reaction mass of 1 M hydrochloric acid.

During the implementation of the task, an unusual method was found for producing 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent-1-en-3-yl chloride] morpholin-4-Ia (I). The method is a two-stage process. The first step is the reaction of 3- (4-oxo-6-chloro-4H-chromen-3-yl) acrylic acid (XII) with 4- (2-methylprop-1-en-1-yl) morpholine (XIII) in ratio of 1: 1.5 in dichloroethane at a temperature of 18-20 ° C. In the second stage, 1 M hydrochloric acid is added to the reaction mass, which leads to the precipitation of the target product.

Presumably at the first stage, the Diels-Alder reaction proceeds with the formation of tetrahydroxantone (XIV). According to available literature, tetrahydroxanthones, similar to compounds (XIV), are unstable and cleave the amine to form 3,4-dihydroxanthones [Bodwell, G.J. Electron deficient dienes 3: rapid access to 2-hydroxybenzophenones via inverse electron demand Diels-Alder-driven domino reactions of a chromone-fused electron deficient diene with enamines / G.J. Bodwell, K.M. Hawco, R.P. da Silva // Synlett. - 2003. - No. 2. - P. 179-182]. However, instead of the expected cleavage of morpholine, a cycle opens, which is a characteristic behavior for Diels-Alder adducts with bulky substituents [Itoh, K. The reaction of b-nitrostyrenes with 2-methoxyfuran: a novel formation of isoxazoline N-oxide together with Michael adducts / K. Itoh, S. Kishimoto // New Journal of Chemistry. - 2000. - V. 24. - P. 347-349]. Disclosure is accompanied by decarboxylation and leads to allylamine (XV). When reacted with hydrochloric acid, this amine forms the corresponding hydrochloride, the target product (I).

The method of producing chloride 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholin-4-Ia (I) studied and carried out in laboratory conditions.

The data of elemental analysis and mass spectrometry, the melting point and the yield of the reaction product are shown in table 1, the spectral characteristics of the obtained compounds are shown in table 2.

The results of a study of anti-tuberculosis activity are presented in FIG. 1-5.

FIG. 1 - MIC of compound (I) in relation to strain H37Rv, where

along the abscissa, the concentration of compound (I), μg / ml; along the ordinate axis, the luminescence intensity of resorufin (a compound formed as a result of the action of living mycobacteria enzymes on resazurin) in conv. units; line 1 - glow in the wells with different concentrations of compound (I), 2 - growth control (glow in the wells without the drug), 3 - growth control 10% of the mycobacteria population; the corresponding dashed lines are the upper and lower boundaries of the confidence interval. MIC = 50 μg / ml.

FIG. 2 - MIC of compound (I) in relation to the clinical strain No. 2067 with multidrug resistance, where

along the abscissa, the concentration of compound (I), μg / ml; along the ordinate axis, the luminescence intensity of resorufin (a compound formed as a result of the action of living mycobacteria enzymes on resazurin) in conv. units; line 1 - luminescence in the wells with different concentrations of compound (I), 2 - growth control (luminescence in the wells without the drug), 3 - growth control 10% of the mycobacteria population, 4 - growth control 1% of the mycobacteria population; the corresponding dashed lines are the upper and lower boundaries of the confidence interval. MIC = 12.5 μg / ml.

FIG. 3 - MIC of compound (I) in relation to clinical strain No. 5227 with multiresistance, where

(see to Fig. 2). MIC = 50.0 μg / ml.

FIG. 4 - MIC of compound (I) in relation to the clinical strain No. 5307 with multiresistance, where:

(see to Fig. 2). MIC = 50.0 μg / ml.

FIG. 5 - MIC of compound (I) in relation to the clinical strain No. 7106 with multidrug resistance, where (see to Fig. 2). MIC = 50.0 μg / ml.

The proposed group of inventions is illustrated by examples of practical implementation.

Example 1. Preparation of 4 - [(1E) -1- (4-oxo-6-chloro-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholin-4-yl chloride ( I)

50 ml of dichloroethane (chemically pure, TU 2631-085-44493179-02) and 5 g (20 mmol) of 3- (4-oxo-6-chloro-4H-chromen-3-yl) acrylic acid are placed in a conical flask then, with stirring, 4.2 g (30 mmol) of 4- (2-methylprop-1-en-1-yl) morpholine are added. The reaction mixture is stirred at a temperature of 18-20 ° C for 3 days. Next, 50 ml of 1 M hydrochloric acid is added to the reaction mass, the resulting emulsion is intensively stirred for 4 hours. The precipitated precipitate of the product is filtered off, washed with dichloroethane and water. The precipitate was recrystallized from water and then dried for 2 hours at 40-45 ° C.

The target product is a white or beige powder. The mass of the product is 5.2 g (68%, based on 3- (4-oxo-6-chloro-4H-chromen-3-yl) acrylic acid).

The composition of the synthesized compound was confirmed by elemental analysis and mass spectrometry data (table. 1). Gross formula: C ₁₉ H ₂₃ Cl ₂ NO ₃ . Found,%: C - 59.14, H - 5.48, N - 3.53. Calculated%: C - 59.38, H - 6.03, N - 3.63. In the mass spectrum (electrospray ionization (ESI), registration of positive ions), peaks of [М-Сl ^- ] ⁺ cations with m / z 348.1343 and 350.1318 with an intensity ratio of 3: 1 are observed, which is associated with the isotopic composition of chlorine.

The structure of the synthesized substance was proved by ¹ H- and ¹³ C-NMR spectroscopy (Table 2).

The NMR spectrum of a ¹ N solution of the test compound in DMSO-d ⁶ in a weak field contains a broadened acid proton singlet in the region of 11.27 ppm. (1H). The proton signals of the aromatic system were recorded as a singlet of 8.72 ppm. (C ² H, 1 H) and peak groups 7.98 ppm. (d, 1H, J 2.3 Hz, C ⁵ N), 7.83 ppm. (dd, 1H, J 2.3 and 9.0 Hz, C ⁷ N), 7.73 ppm. (d, 1H, J 9.0 Hz, C ⁸ N). Signals 6.81 ppm (dd, 1H, J 10.3 and 15.8 Hz) and 6.57 ppm. (d, 1H, J 15.8 Hz) correspond to olefin protons. The spin-spin interaction constant of 15.8 Hz indicates the trans configuration of the double bond. In a strong field is a series of multiplet signals of protons of the morpholine cycle of 4.07 ppm. (2H), 3.92 ppm. (2H), 3.44 ppm. (2H) and 3.07 ppm. (2H) and CH-N groups 3.75 ppm. (1H). The isopropyl fragment is detected as a multiplet of 2.42 ppm. (1H) and two doublets of methyl groups 1.06 ppm. (3H, J 6.8 Hz) and 0.93 ppm. (3H, J 6.8 Hz).

The ¹³ C NMR spectrum in a weak field contains signals of carbon atoms C ⁴ = O (174.7 ppm) and C _sp2- O (157.2 and 154.3 ppm). Signals with chemical shifts of 134.6, 130.6, 130.1, 125.0, 124.6, 122.9, 121.5 and 119.6 ppm. belong to sp ² -hybrid carbon atoms of the molecule. The signals of the carbon atoms of the morpholine cycle and CH-N have a chemical shift of 75.0, 63.4, 63.3, 50.7 and 49.9 ppm. The isopropyl fragment is recorded as peaks of 26.5, 21.0 and 17.2 ppm.

Example 2. Tuberculostatic activity of chloride 4 - [(1E) -1- (4-oxo-6-chloro-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholin-4-ya (I) (in vitro studies)

The study of tuberculostatic activity was carried out by the method of double serial micro-dilutions in Middlebrook’s liquid synthetic medium (Middlebrook 7Н9 BrothBase, Sigma-Aldrich, cat. No. MO 178) with 10% OADC growth supplement (BBL Middlebrook OADCEnrichment, Becton Dickinson, cat. No. 245116) a well plate showing mycobacteria growth using 0.01% resazurin (Resazurinsodiumsalt, Sigma, Cat. No. R7017) (REMA method - Resazurin Microdilution Assay; Palomino J.-C., Martin A., Camacho M., Guerra H., Swings J., Portaels F. Resazurin Microtiter Assay Plate Simple and Inexpensive method for Detection of Dgug Resistance in Mycobacterium tuberculosis. // Antimicrob. Agents Chemotherap. - 2002. - Vol. 46. - No. 8. - P. 2720-2722) and registration grew and using a tablet fluorimeter FLUO Star Optima (excitation wavelength - 520 nm radiation - 590 nm). As test strains used:

1. Mycobacterium tuberculosis H ₃₇ Rv strain sensitive to anti-tuberculosis drugs (TVS # 1/47, source - Institute of Hygiene and Epidemiology, Prague, 1976, obtained from the Federal State Budgetary Institution Scientific Center for Expert Advice of Medical Applications of the Russian Ministry of Health);

2. 4 clinical strains isolated from patients with newly diagnosed pulmonary tuberculosis (collection of the St. Petersburg Research Institute of Phthisiopulmonology of the Russian Ministry of Health):

- clinical strain No. 2067 with multidrug resistance, resistance spectrum - isoniazid, rifampicin, kanamycin, mutations - in the ahp C gene (T10), in the rpoB gene (His526 → Arg);

- clinical strain No. 5227 with multi-resistance, resistance spectrum: isoniazid, streptomycin; mutations - in the kat G gene (Ser315 → Thr), ahp C (A 10);

- clinical strain No. 5307 with multiresistance, resistance spectrum: isoniazid, streptomycin, ethionamide; mutation - in the inhA gene (T15);

- clinical strain No. 7106 with multidrug resistance, resistance spectrum: isoniazid, streptomycin, rifampicin, ethambutol, kanamycin, ethionamide, capreomycin, amikacin; mutations - in the katG gene (Ser315 → Thr), in the rpoB gene (Ser531 → Leu).

The minimum inhibitory concentration (MIC) of compound (I) in relation to the H37Rv strain sensitive to anti-TB drugs, as well as drug-resistant clinical isolates Nos. 5227, 5307, 7106 was 50 μg / ml, in relation to clinical strain No. 2067 - 12.5 μg / ml (Fig. 1-5).

The advantage of compound (I) is that it has a similar degree of tuberculostatic activity both in relation to sensitive and drug-resistant strains, regardless of the nature and spectrum of their resistance to anti-TB drugs, or from the set of mutations associated with resistance to isoniazid and rifampicin - the main anti-TB drugs.

Thus, 4 - [(1E) -1- (4-oxo-6-chloro-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholin-4-yl chloride (I ) possesses tuberculostatic activity against drug-resistant clinical strains of Mycobacterium tuberculosis and is promising for use in medical practice.

Claims (6)

1. Chloride 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholin-4-th formula I

2. The compound according to claim 1, having anti-tuberculosis activity. 3. The method of producing chloride 4 - [(1E) -1- (6-chloro-4-oxo-4H-chromen-3-yl) -4-methylpent-1-en-3-yl] morpholin-4-Ia of the formula I

by reacting 3- (4-oxo-6-chloro-4H-chromen-3-yl) acrylic acid with 4- (2-methylprop-1-en-1-yl) morpholine in a ratio of 1: 1.5 in dichloroethane at a temperature of 18-20 ° C, and the target product is obtained in the form of a precipitate upon exposure to the reaction mass of 1 M hydrochloric acid.

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