WO2017181948A1

WO2017181948A1 - Substituted oxazolidone and application thereof

Info

Publication number: WO2017181948A1
Application number: PCT/CN2017/081010
Authority: WO
Inventors: 王义汉; 邢青峰
Original assignee: 深圳市塔吉瑞生物医药有限公司
Priority date: 2016-04-22
Filing date: 2017-04-19
Publication date: 2017-10-26
Also published as: CN108368139A; CN108368139B

Abstract

An oxazolidone compound represented by formula (I), or a crystalline form, pharmaceutically acceptable salt, prodrug, metabolite, stereoisomer, isotopic isomer, hydrate, or solvate thereof, and a pharmaceutical composition comprising the compound. The compound has demonstrated inhibitory activity against a broad spectrum of bacteria and low toxicity, and can be used to prepare an antibiotic.

Description

Substituted oxazolidinone compounds and their applications

Technical field

The invention belongs to the field of medicine. In particular, the present invention relates to deuterated oxazolidinone derivatives and uses thereof, and more particularly to oxazolidinone compounds which are useful as antibiotics.

Background technique

Antibiotics are one of the most frequently used essential drugs in the clinic. The irregular use of antibiotics due to historical reasons has caused the bacteria to be severely resistant to existing antibiotics. Multidrug-resistant (MDR) bacterial infections have become one of the major threats to public health worldwide. Because of multi-drug resistance, the combination of antibiotics is increasing, and drug-drug interactions also increase the risk of adverse reactions. According to the 2013 National Annual Report on Adverse Drug Reaction Monitoring released by the State Food and Drug Administration, the adverse reactions caused by antibiotics rank first in the adverse reactions/incident reports. The "super bacteria" that are resistant to most of the available antibiotics are also spreading around the world at an alarming rate. On the other hand, the development of new and effective antibiotics has not increased with the increase of MDR bacteria. According to the US Center for New Drug Evaluation (CDER), the number of newly approved antibiotics has been decreasing since 1980. Existing antibiotics have been unable to cure the increasing infection and resistance.

Sivextro (common name: tedizolid phosphate, formerly known as TR-701) was developed by Cubist Pharmaceuticals Inc. and is an oxazolidinone antibiotic. Sivextro is a prodrug that is rapidly converted to a biologically active tedizolid by phosphatase in the body. The latter binds to the ribosomal 50S subunit of the bacterium, thereby inhibiting protein synthesis. Although at least 10 similar compounds have entered the clinic since Pfizer's similar antibiotic linezolid was approved by the US FDA in 2000, Sivextro is the first second-generation oxazolidinone antibiotic to be approved by the FDA. Compared with the first-generation product linezolid, Sivextro has 2-8 times higher inhibitory activity against some bacteria in vitro, and the safety is also improved to some extent.

Deuterated modification is a potentially attractive strategy for improving the metabolic properties of drugs. Helium is a safe, stable, non-radiative isotope of hydrogen. Compared with the C-H bond, the C-D bond formed by ruthenium and carbon is stronger because of the lower vibration frequency. In addition, the "heavy hydrogen" version of the drug may be more stable to degradation and longer in the body. Deuterated drugs have a positive impact on safety, efficacy and tolerance, and have excellent research prospects.

Summary of the invention

In view of the above technical problems, the present invention discloses an oxazolidinone compound and a composition comprising the same as an effective antibacterial active compound and/or having better pharmacodynamic/pharmacokinetic properties.

In this regard, the technical solution adopted by the present invention is as follows:

In a first aspect of the invention, there is provided an oxazolidinone compound of the formula (I), or a crystalline form thereof, a pharmaceutically acceptable salt, a prodrug, a metabolite, a stereoisomer or an isotope Body, hydrate or solvent compound:

In the formula:

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently hydrogen, deuterium or halogen. Or trifluoromethyl;

With the proviso that at least one of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ is 氘Generation or embarrassment.

As a further improvement of the present invention, each of R ¹ , R ² and R ^{3 is} independently hydrazine or hydrogen.

As a further improvement of the present invention, R ⁴ , R ⁵ and R ⁶ are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ⁷ , R ⁸ and R ⁹ are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ¹⁰ , R ¹¹ and R ¹² are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ¹³ and R ¹⁴ are each independently hydrazine or hydrogen.

In another preferred embodiment, the compound is selected from the group consisting of the compounds or pharmaceutically acceptable salts thereof, but is not limited to the following compounds:

In another preferred embodiment, the cerium isotope content of the cerium in the deuterated position is at least greater than the natural strontium isotope content (0.015%), preferably greater than 30%, more preferably greater than 50%, and even more preferably greater than 75%. Preferably, the ground is greater than 95%, more preferably greater than 99%.

Specifically, in the present invention, each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ The strontium isotope content in the deuterated position is at least 5%, preferably greater than 10%, more preferably greater than 15%, more preferably greater than 20%, more preferably greater than 25%, still more preferably greater than 30%, more preferably More than 35%, more preferably more than 40%, more preferably more than 45%, more preferably more than 50%, more preferably more than 55%, more preferably more than 60%, more preferably more than 65%, more preferably more than 70%, more preferably more than 75%, more preferably more than 80%, more preferably more than 85%, more preferably more than 90%, more preferably more than 95%, more preferably more than 99%.

In another preferred embodiment, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² of the compound of formula (I), R ¹³ and R ¹⁴ , at least one of R contains ruthenium, more preferably two R contains ruthenium, more preferably three R contains ruthenium, more preferably four R contains ruthenium, more preferably five R contains ruthenium, more preferably five R-containing ruthenium, more preferably Preferably, the six R-containing yttrium, more preferably the seven R-containing yttrium, more preferably eight R-containing yttrium, more preferably nine R-containing yttrium, more preferably ten R-containing yttrium, more preferably eleven R contains hydrazine, more preferably twelve R 氘, more preferably thirteen R 氘, more preferably fourteen R 氘.

In another preferred embodiment, the compound does not include a non-deuterated compound.

In a second aspect of the invention, there is provided a method of preparing a pharmaceutical composition comprising the steps of: pharmaceutically acceptable carrier and a compound of the first aspect of the invention, or a crystalline form thereof, pharmaceutically acceptable The accepted salt, hydrate or solvate is mixed to form a pharmaceutical composition.

In a third aspect of the invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of the first aspect of the invention, or a crystalline form thereof, a pharmaceutically acceptable salt, hydrated Or a solvate.

The oxazolidinone compound of the present invention exhibits inhibitory activity against a broad spectrum of bacteria, methicillin-resistant Staphylococcus aureus, and vancomycin-resistant Enterococcus, and has relatively excellent antibacterial activity at a relatively low concentration or in vivo. .

Further, the compound of the present invention can be expressed to include Gram-positive bacteria such as Staphylococcus, Enterococcus and Streptococcus, anaerobic microorganisms such as bacteroids and Clostridium, and acid-tolerant microorganisms such as Mycobacterium tuberculosis and Mycobacterium avium. The potent antibacterial activity of human and animal pathogens.

The composition of the present invention may comprise at least one active ingredient having a function similar to an oxazolidinone derivative.

For formulating the pharmaceutical composition, at least one compound of formula (I) can be combined with at least one pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers can include physiological saline, sterile water, Ringer's solution, physiological saline buffer solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like.

The pharmaceutical composition may contain conventional excipients such as antioxidants, buffers, soil cleaners and the like. The composition is also mixed with a diluent, a diintegrant, a surfactant, a binder, a lubricant, an aqueous solution, a suspension or the like to form an injection, a powder, a capsule, a granule, a tablet or the like. Preferably, the formulation can be prepared by the method described in Remington's Pharmaceutical Science (latest edition) (Mack Publishing Company, Easton PA, etc.) depending on the disease or component.

The compounds of the invention may be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally, topically, and the like. Chemical The dosage of the compound can vary depending on the particular compound employed, the mode of administration, the condition and severity of the condition to be treated, and the various physical factors associated with the subject being treated. When the compound of the present invention is administered to an individual at a daily dose of about 8 to 30 mg, preferably 12 to 21 mg per kg of body weight, satisfactory results can be obtained according to the use of the present invention. More preferably, the above daily dose is divided into several doses per day.

The oxazolidinone derivative of the present invention exhibits inhibitory activity and low toxicity against a broad spectrum of bacteria. A prodrug prepared by reacting a compound having a hydroxyl group with an amino acid or a phosphate has high water solubility.

Further, the derivatives of the present invention may be shown to include Gram-positive bacteria such as Staphylococcus, Enterococcus and Streptococcus, anaerobic microorganisms such as bacteroids and Clostridium, and acid-tolerant microorganisms such as Mycobacterium tuberculosis, Mycobacterium avium The potent antibacterial activity of human and animal pathogens.

Therefore, a composition containing the oxazolidinone derivative is used in an antibiotic.

The invention also includes isotopically labeled compounds, equivalent to the original compounds disclosed herein. Examples of isotopes which may be listed as compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine isotopes such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, ^{31, respectively.} P, ³² P, ¹⁸ F and ³⁶ Cl. a compound, or an enantiomer, a diastereomer, an isomer, or a pharmaceutically acceptable salt or solvate of the present invention, wherein an isotope or other isotopic atom containing the above compound is within the scope of the present invention . Certain isotopically-labeled compounds of the present invention, such as the radioisotopes of ³ H and ¹⁴ C, are also among them, useful in tissue distribution experiments of drugs and substrates.氚, ie ³ H and carbon-14, ie ¹⁴ C, are easier to prepare and detect and are preferred in isotopes. Isotopically labeled compounds can be prepared in a conventional manner by substituting a readily available isotopically labeled reagent with a non-isotopic reagent using the protocol of the examples.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

Compared with the prior art, the beneficial effects of the present invention are: the substituted oxazolidinone compound disclosed by the present invention and the composition comprising the same for the spectrum bacteria (including Gram-positive bacteria such as Staphylococcus, Enterococcus) With Streptococcus, anaerobic microorganisms such as bacteriophages and Clostridium and acid-tolerant microorganisms such as Mycobacterium tuberculosis, Mycobacterium avium, have excellent inhibition and have better pharmacokinetic parameter characteristics. The dosage can be varied and a long acting formulation can be formed to improve suitability. Replacing a hydrogen atom in a compound with hydrazine can increase the drug concentration of the compound in an animal to improve the efficacy of the drug due to its strontium isotope effect. Substitution of a hydrogen atom in a compound with hydrazine may increase the safety of the compound due to inhibition of certain metabolites.

detailed description

The preparation of the structural compound of the formula I of the present invention is more specifically described below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention may also be conveniently prepared by combining various synthetic methods described in the specification or known in the art, and such combinations are readily made by those skilled in the art to which the present invention pertains.

Example 1 Preparation of (R)-3-(4-(2-(2-d3-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyl Azole Alkan-2-ketophosphate (Compound T-1)

The specific synthesis steps are as follows:

Step 1 Synthesis of 2-(2-d3-methyltetrazol-5-yl)-5-bromopyridine (Compound 2).

5-Bromo-2-(2H-tetrazol-5-yl)pyridine (0.5 g, 2.2 mmol) and K ₂ CO ₃ (0.61 g, 4.42 mmol) were dissolved in 10 mL DMF and slowly added dropwise under ice bath. Methyl iodide (0.42 g, 2.88 mmol). The mixture was stirred for 1 hour in an ice bath, and the material disappeared by TLC. 40 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. EtOAc (EtOAc) -5-yl)-5-bromopyridine (Compound 2) 0.27 g, yield 52.5%. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.83 (dd, J = 2.4, 0.8 Hz, 1H), 8.14 (dd, J = 8.4, 0.8 Hz, 1H), 8.00 (dd, J = 8.4, 2.3 Hz, 1H). ESI-MS: 243 [M ⁺ +1].

Step 2(R)-3-(3-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)- Synthesis of 5-hydroxymethyloxazolidin-2-one (Compound 4).

(R)-3-(4-Bromo-3-fluorophenyl)-5-hydroxymethyl-oxazolidin-2-one (0.58 g, 2.0 mmol), bis-pinacol borate (0.66 g, 2.6 mmol), potassium acetate (0.29 g, 3.0 mmol) and Pd(PPh ₃ ) ₂ Cl ₂ (0.07 g, 0.1 mmol) were added to a 50 mL two-necked flask, 30 ml of dioxane was added, and three times with nitrogen, 90 ° C The reaction was overnight. The reaction solution was cooled to room temperature, then added with water (60 mL), EtOAc (EtOAc) Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-5-hydroxymethyloxazolidine-2- Ketone (Compound 4) 0.55 g, yield 81.6%. ESI-MS: 338 [M ⁺ +1].

Step 3(R)-3-(4-(2-(2-(methyl-d3)tetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyl Synthesis of oxazolidine-2-one (Compound 5).

2-(2-(Methyl-d3)tetrazol-5-yl)-5-bromopyridine (0.15 g, 0.6 mmol), (R)-3-(3-fluoro-4-(4,4, 5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-5-(hydroxymethyl)oxazolidin-2-one (0.21 g, 0.6 mmol) Potassium phosphate (0.27 g, 1.3 mmol) and Pd(dppf)Cl ₂ (0.045 g, 0.06 mmol) were dissolved in 20 mL of dioxane, 2 mL of water was added, and the nitrogen ball was replaced three times, and reacted at 90 ° C overnight. The reaction solution was cooled to room temperature, and then added with 50 mL of water, EtOAc (EtOAc) 2-(2-(Methyl-d3)tetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one (Compound 5) 0.15 g The yield was 64.8%. ^{1 H NMR (400MHz, DMSO-} d6) δ8.94 (s, 1H), 8.26-8.17 (m, 2H), 7.78-7.68 (m, 2H), 7.53 (dd, J = 8.6,2.3Hz, 1H) , 5.25 (q, J = 5.5 Hz, 1H), 4.77 (ddt, J = 9.6, 6.4, 3.5 Hz, 1H), 4.16 (t, J = 9.1 Hz, 1H), 3.91 (dd, J = 9.0, 6.1 Hz, 1H), 3.71 (ddd, J = 12.4, 5.5, 3.3 Hz, 1H), 3.59 (ddd, J = 12.4, 5.8, 3.9 Hz, 1H). ESI-MS: 374 [M ⁺ +1].

Step 4(R)-3-(4-(2-(2-(methyl-d3)tetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyl Synthesis of oxazolidine-2-one phosphate (compound).

(R)-3-(4-(2-(2-(methyl-d3)tetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazole Alkan-2-one (0.15 g, 0.4 mmol) and triethylamine (0.25 g, 2.4 mmol) were dissolved in 20 mL of tetrahydrofuran, and phosphorus oxychloride (0.37 g, 2.43 mmol) was slowly added in an ice bath and reacted in an ice bath. After 3 hours, 5 mL of water was added and stirring was continued for 1 hour. Most of the tetrahydrofuran was distilled off, and 30 mL of a saturated sodium carbonate solution was added to adjust pH = 10, and washed with dichloromethane, and then the aqueous phase was adjusted to pH 1-2 with 3 mol/L hydrochloric acid, and a large amount of solid was precipitated. Filtration, washing the cake with water and drying to give (R)-3-(4-(2-(2-(methyl-d3)tetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl) -5-Hydroxymethyloxazolidin-2-one phosphate 45 mg, yield 24.7%. ^{1 H NMR (300MHz, DMSO-} d6) δ8.94 (s, 1H), 8.22 (d, 2H), 7.71 (d, J = 19.7Hz, 2H), 7.52 (s, 1H), 4.95 (m, 1H ), 3.92-4.23 (m, 4H). ESI-MS: 454 [M ⁺ +1].

Example 2 Preparation of (R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl-3,4,6-d3)-3-fluorophenyl)- 5-hydroxymethyl Oxazolidin-2-one phosphate (Compound T-2)

The specific synthesis steps are as follows:

Step 1 Synthesis of 2-(2-methyltetrazol-5-yl)-5-bromopyridine (Compound 6).

5-Bromo-2-(2H-tetrazol-5-yl)pyridine (0.5 g, 2.2 mmol) and K ₂ CO ₃ (0.61 g, 4.42 mmol) were dissolved in 10 mL of DMF. (0.41 g, 2.88 mmol). The mixture was stirred for 1 hour in an ice bath, and the material disappeared by TLC. 40 mL of water was added to the reaction solution, and the mixture was extracted with EtOAc. EtOAc (EtOAc) ) 5-bromopyridine 0.28 g, yield 52.7%. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.83 (d, J = 2.3 Hz, 1H), 8.14 (dd, J = 8.4, 0.7 Hz, 1H), 8.00 (dd, J = 8.4, 2.3 Hz, 1H), 4.45 (s, 3H). ESI-MS: 240 [M ⁺ +1].

Step 2 Synthesis of 2-(2-methyltetrazol-5-yl)-5-bromo-3,4,6-d3-pyridine (Compound 7).

2-(2-Methyltetrazol-5-yl)-5-bromopyridine (0.28 g, 1.17 mmol) was added to a 15 mL aqueous solution of 10% sodium sulphoxide and reacted at 180 ° C for 5 hours under nitrogen atmosphere. After cooling to room temperature, 10 mL of water was added, and the mixture was evaporated. ¹ H NMR (300 MHz, DMSO-d6) δ 4.44 (s, 3H). ESI-MS: 243 [M ⁺ +1].

Step 3(R)-3-(4-(2-(2-Methyltetrazol-5-yl)pyridin-5-yl-3,4,6-d3)-3-fluorophenyl)-5- Synthesis of hydroxymethyloxazolidin-2-one (Compound 8).

2-(2-Methyltetrazol-5-yl)-5-bromo-3,4,6-d3-pyridine (0.15 g, 0.6 mmol), (R)-3-(3-fluoro-4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-5-(hydroxymethyl)oxazolidin-2-one ( 0.21 g, 0.6 mmol), potassium phosphate (0.27 g, 1.3 mmol) and Pd(dppf)Cl ₂ (0.045 g, 0.06 mmol) were dissolved in 20 mL of dioxane, 2 mL of water was added, and the nitrogen ball was replaced three times, 90 ° C The reaction was overnight. The reaction solution was cooled to room temperature, and then added with 50 mL of water, EtOAc (EtOAc) 2-(2-Methyltetrazol-5-yl)pyridin-5-yl-3,4,6-d3)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one 0.15g The yield was 64.8%. ^{1 H NMR (300MHz, DMSO-} d6) δ7.82-7.66 (m, 2H), 7.53 (d, J = 9.1Hz, 1H), 5.27 (t, J = 5.6Hz, 1H), 4.75 (d, J = 9.0 Hz, 1H), 4.48 (s, 3H), 4.16 (t, J = 9.1 Hz, 1H), 3.91 (t, J = 7.5 Hz, 1H), 3.75-3.53 (m, 2H). ESI-MS: 374 [M ⁺ +1].

Step 4(R)-3-(4-(2-(2-Methyltetrazol-5-yl)pyridin-5-yl-3,4,6-d3)-3-fluorophenyl)-5- Synthesis of hydroxymethyloxazolidine-2-one phosphate (Compound T-2).

(R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl-3,4,6-d3)-3-fluorophenyl)-5-hydroxy Methyloxazolidin-2-one (0.15 g, 0.4 mmol) and triethylamine (0.25 g, 2.4 mmol) were dissolved in 20 mL of tetrahydrofuran, and phosphorus oxychloride (0.37 g, 2.43 mmol) was slowly added in an ice bath. After reacting for 3 hours in an ice bath, 5 mL of water was added and stirring was continued for 1 hour. Most of the tetrahydrofuran was distilled off, and 30 mL of a saturated sodium carbonate solution was added to adjust pH = 10, and washed with dichloromethane, and then the aqueous phase was adjusted to pH 1-2 with 3 mol/L hydrochloric acid, and a large amount of solid was precipitated. Filtration, filter cake washed with water and dried to give (R)-3-(4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl-3,4,6-d3)-3- Fluorophenyl)-5-hydroxymethyloxazolidin-2-one phosphate 46 mg, yield 25%. ^{1 H NMR (400MHz, DMSO-} d6) δ7.80-7.67 (m, 2H), 7.52 (dd, J = 8.7,2.2Hz, 1H), 4.96 (m, 1H), 4.48 (s, 3H), 4.23 (t, J = 9.1 Hz, 1H), 4.18-3.99 (m, 2H), 3.96-3.88 (m, 1H). ESI-MS: 454 [M ⁺ +1].

Biological activity test

(1) Evaluation of antibacterial activity in vivo.

The antibacterial activity of the oxazolidinone compound was tested by the method described in Chemotheraphy, 29(1), 76, (1981). The dilute solution of agar was used to include methicillin-resistant Staphylococcus aureus (MRSA) and resistant to vancomycin. The antibacterial activity of the enterococci (VRE) is expressed as the minimum inhibitory concentration (MIC50, μg/m Rise). The results are shown in Table 1 below.

Table 1 Example compound antibacterial activity test results

化合物Compound	MRSA最小抑制浓度(MIC50，μM)MRSA minimum inhibitory concentration (MIC50, μM)	VRE最小抑制浓度(MIC50，μM)VRE minimum inhibitory concentration (MIC50, μM)
T-1T-1	<1<1	<1<1
T-2T-2	<1<1	<1<1

The experimental results show that the compound of the present invention exhibits sufficient antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Thus, the compounds of the invention are useful as antibiotics.

(2) Pharmacokinetic evaluation in rats

Eight male Sprague-Dawley rats, 7-8 weeks old, weighing 210 g, were divided into 2 groups, 4 in each group, and a single oral dose of 5 mg/kg (a) control group: (R)-3-( 4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one; (b) Test group: Example compounds were compared for differences in pharmacokinetics.

Rats were fed a standard diet and given water. Fasting began 16 hours before the test. The drug was dissolved with PEG400 and dimethyl sulfoxide. Blood was collected from the eyelids at a time point of 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours after administration.

Rats were briefly anesthetized after inhalation of ether, and 300 μL of blood samples were collected from the eyelids in test tubes. There was 30 μL of 1% heparin salt solution in the test tube. The tubes were dried overnight at 60 ° C before use. After the blood sample collection was completed at a later time point, the rats were anesthetized with ether and sacrificed.

Immediately after the blood sample is collected, gently invert the tube at least 5 times to ensure adequate mixing and place on ice. Blood samples were centrifuged at 5000 rpm for 5 minutes at 4 ° C to separate plasma from red blood cells. Pipette 100 μL of plasma into a clean plastic centrifuge tube, indicating the name and time of the compound. Plasma was stored at -80 °C prior to analysis. The concentration of the compound of the invention in plasma was determined by LC-MS/MS. Pharmacokinetic parameters were calculated based on the plasma concentration of each animal at different time points.

The experimental results show that the compound of the present invention has better pharmacokinetics in animals relative to the control compound, and thus has better pharmacodynamics and therapeutic effects.

(3) Metabolic stability evaluation

Microsomal experiments: human liver microsomes: 0.5 mg/mL, BD Gentest; rat liver microsomes: 0.5 mg/mL, Xenotech; mouse liver microsomes: 0.5 mg/mL, Xenotech; coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

Preparation of stock solution: A certain amount of the compound powder of the example was accurately weighed and dissolved to 5 mM with DMSO.

Preparation of phosphate buffer (100 mM, pH 7.4): Mix 150 mL of pre-formed 0.5 M potassium dihydrogen phosphate and 700 mL of 0.5 M potassium dihydrogen phosphate solution, and adjust the mixture with 0.5 M potassium dihydrogen phosphate solution. The pH was adjusted to 7.4, diluted 5 times with ultrapure water before use, and magnesium chloride was added to obtain a phosphate buffer (100 mM) containing 100 mM potassium phosphate, 3.3 mM magnesium chloride, and a pH of 7.4.

A solution of NADPH regeneration system (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was prepared and placed on wet ice before use.

Formulation stop solution: acetonitrile solution containing 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard). Take 25057.5 μL of phosphate buffer (pH 7.4) into a 50 mL centrifuge tube, add 812.5 μL of human liver microsomes, and mix to obtain a liver microsome dilution with a protein concentration of 0.625 mg/mL. Take 25057.5 μL of phosphate buffer (pH 7.4) into a 50 mL centrifuge tube, add 812.5 μL of rat liver microsomes, and mix to obtain a liver microsome dilution with a protein concentration of 0.625 mg/mL. 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, and 812.5 μL of mouse liver microsomes were added and mixed to obtain a liver microsome dilution having a protein concentration of 0.625 mg/mL.

Incubation of the sample: The stock solution of the corresponding compound was diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile as a working solution, and was used. 398 μL of human liver microsomes or rat liver microsomes or mouse liver microsome dilutions were added to 96-well incubation plates (N=2), and 2 μL of 0.25 mM working solution was added and mixed.

Determination of metabolic stability: 300 μL of pre-cooled stop solution was added to each well of a 96-well deep well plate and placed on ice as a stop plate. The 96-well incubation plate and the NADPH regeneration system were placed in a 37 ° C water bath, shaken at 100 rpm, and pre-incubated for 5 min. 80 μL of the incubation solution was taken from each well of the incubation plate, added to the stopper plate, and mixed, and 20 μL of the NADPH regeneration system solution was added as a sample of 0 min. Then, 80 μL of the NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and start timing. The corresponding compound had a reaction concentration of 1 μM and a protein concentration of 0.5 mg/mL. 100 μL of the reaction solution was taken at 10, 30, and 90 min, respectively, and added to the stopper, and the reaction was terminated by vortexing for 3 min. The plate was centrifuged at 5000 x g for 10 min at 4 °C. 100 μL of the supernatant was taken into a 96-well plate to which 100 μL of distilled water was previously added, mixed, and sample analysis was performed by LC-MS/MS.

Data analysis: The peak area of the corresponding compound and the internal standard was detected by LC-MS/MS system, and the ratio of the peak area of the compound to the internal standard was calculated. The slope is measured by the natural logarithm of the percentage of the remaining amount of the compound versus time, and t _1/2 and CL _{int are} calculated according to the following formula, where V/M is equal to 1/protein concentration.

The Tedizolid, Compounds T-1 and T-2 were analyzed according to the above procedure, and the results are shown in Table 2.

Table 2 Determination of metabolic stability of the compound of the example

As shown in Table 2, the compounds of the present invention exhibited excellent metabolic stability in human liver microsomes, rat liver microsomes, and mouse liver microsome experiments, and were significantly superior to the undeuterated compound Tedizolid. This demonstrates that the deuterated compounds T-1 and T-2 of the present invention significantly improve the metabolic stability of the non-deuterated compounds.

It is to be understood that the examples are merely illustrative of the invention and are not intended to limit the scope of the invention, and the experimental methods in which the specific conditions are not indicated in the examples, usually in accordance with conventional conditions or in accordance with the conditions suggested by the manufacturer. Parts and percentages are parts by weight and percentage by weight unless otherwise stated.

The above is a further detailed description of the present invention in connection with the specific preferred embodiments, and the specific embodiments of the present invention are not limited to the description. It will be apparent to those skilled in the art that the present invention may be made without departing from the spirit and scope of the invention.

Claims

A substituted oxazolidinone compound characterized by having an oxazolidinone compound of the formula (I), or a crystalline form thereof, a pharmaceutically acceptable salt, a prodrug, a metabolite, a stereoisomer Isotope variant, hydrate or solvent compound,

Wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 and R 14 are each independently hydrogen and deuterium. , halogen or trifluoromethyl;

With the proviso that at least one of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 is 氘Generation or embarrassment.
The oxazolidinone compound according to claim 1, wherein each of R 1 , R 2 and R 3 is independently hydrazine or hydrogen.
The oxazolidinone compound according to claim 1, wherein each of R 4 , R 5 and R 6 is independently hydrazine or hydrogen.
The oxazolidinone compound according to claim 1, wherein each of R 7 , R 8 and R 9 is independently hydrazine or hydrogen.
The oxazolidinone compound according to claim 1, wherein each of R 10 , R 11 and R 12 is independently hydrazine or hydrogen.
The oxazolidinone compound according to claim 1, wherein each of R 13 and R 14 is independently hydrazine or hydrogen.
The oxazolidinone compound according to claim 1, wherein the compound is selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof, but is not limited to the following compounds:
A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a substituted oxazolidinone compound according to any one of claims 1 to 7, or a crystalline form thereof, a pharmaceutically acceptable salt thereof A pharmaceutical composition of a hydrate or solvate, stereoisomer, prodrug, metabolite or isotopic variant is used as an active ingredient and contains a conventional pharmaceutical carrier.
Use of an oxazolidinone compound according to any one of claims 1 to 7, characterized in that as a spectrobacterium, including Gram-positive bacteria such as Staphylococcus, Enterococcus and Streptococcus, anaerobic microorganisms Antibiotics such as bacteroids and Clostridium and acid-resistant microorganisms such as tuberculosis, avian mycobacteria, and human pathogens.