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Berg, Kaja ; Hegde, Pooja ; Pujari, Venugopal , et al. Eur. J. Med. Chem.,2023,249,115125. DOI: 10.1016/j.ejmech.2023.115125 PubMed ID: 36682292
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Abstract: The electron transport chain (ETC) in the cell membrane consists of a series of redox complexes that transfer electrons from electron donors to acceptors and couples this electron transfer with the transfer of protons (H+) across a membrane. This process generates proton motive force which is used to produce ATP and a myriad of other functions and is essential for the long-term survival of Mycobacterium tuberculosis (Mtb), the causative organism of tuberculosis (TB), under the hypoxic conditions present within infected granulomas. Menaquinone (MK), an important carrier molecule within the mycobacterial ETC, is synthesized de novo by a cluster of enzymes known as the classic/canonical MK biosynthetic pathway. MenA (1,4-dihydroxy-2-naphthoate prenyltransferase), the antepenultimate enzyme in this pathway, is a verified target for TB therapy. In this study, we explored structure-activity relationships of a previously discovered MenA inhibitor scaffold, seeking to improve potency and drug disposition properties. Focusing our campaign upon three molecular regions, we identified two novel inhibitors with potent activity against MenA and Mtb (IC50 = 13-22 μM, GIC50 = 8-10 μM). These analogs also displayed substantially improved pharmacokinetic parameters and potent synergy with other ETC-targeting agents, achieving nearly complete sterilization of Mtb in combination therapy within two weeks in vivo. These new inhibitors of MK biosynthesis present a promising new strategy to curb the continued spread of TB.
Keywords: 1,4-dihydroxy-2-naphthoate prenyltransferase ; MenA ; MenA inhibitors ; Menaquinone ; Mtb ; Mycobacterium tuberculosis ; Piperidine derivatives ; SAR
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CAS No. : | 161975-39-9 | MDL No. : | MFCD02082459 |
Formula : | C12H23NO5S | Boiling Point : | - |
Linear Structure Formula : | - | InChI Key : | RXNQBVRCZIYUJK-UHFFFAOYSA-N |
M.W : | 293.38 | Pubchem ID : | 2765838 |
Synonyms : |
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Chemical Name : | tert-Butyl 4-(((methylsulfonyl)oxy)methyl)piperidine-1-carboxylate |
Signal Word: | Warning | Class: | N/A |
Precautionary Statements: | P261-P280-P301+P312-P302+P352-P305+P351+P338 | UN#: | N/A |
Hazard Statements: | H302-H315-H319-H335 | Packing Group: | N/A |
GHS Pictogram: |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With N,N,N',N'-tetramethylguanidine; In tetrahydrofuran; N,N-dimethyl-formamide; for 24h;Heating / reflux; | 2.2. 1,1-dimethylethyl 4-[(2,4-dioxo-1,3-oxazolidin-3-yl)methyl]piperidine-1-carboxylate A suspension of 13.60 g (46.36 mmol) of 1,1-dimethylethyl 4-[(methylsulphonyl)oxy]methyl}piperidine-1-carboxylate, prepared in step 2.1., 9.37 g (92.72 mmol) of 1,3-<strong>[2346-26-1]oxazolidine-2,4-dione</strong> and 16.02 g (139.08 mmol) of 1,1,3,3-tetramethylguanidine in a mixture of 180 ml of tetrahydrofuran and 30 ml of dimethylformamide is heated at reflux for 24 hours. It is allowed to return to ambient temperature and is concentrated under reduced pressure. The residue is taken up in dichloromethane and water and the aqueous phase is separated off and extracted twice with dichloromethane. The combined organic phases are washed with saturated aqueous sodium chloride solution and dried over sodium sulphate. Following evaporation of the solvent, the residue obtained is purified by chromatography on silica gel, eluding with a 98/2 then 95/5 mixture of dichloromethane and methanol. This gives 12.53 g of product in the form of an orange-brown solid. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
95.05% | With potassium carbonate; In N,N-dimethyl-formamide; at 80℃; for 16h;Inert atmosphere; | Under nitrogen atmosphere, potassium carbonate (74.12 g, 536.28 mmol) was added into a solution of intermediate 7A (94.40 g, 321.77 mmol) and <strong>[4983-28-2]2-chloropyrimidin-5-ol</strong> (35.00 g, 268.14 mmol) in DMF (1.00 L). The reaction solution was left at 80 C. for 16 hours, and thin layer chromatography was used to detect the completion of the reaction. Then the reaction solution was cooled to room temperature and concentrated, then water (500 mL) was added into the residue and extracted with ethyl acetate (300 mL*3). The organic phase was washed with brine (400 mL*2) and dried over anhydrous sodium sulfate, then filtered and concentrated. Then the residue was purified by column chromatography to give the intermediate 7B (pale yellow solid, 84.00 g, 95.05% yield). LCMS (ESI) m/z: 327.7 (M+1). 1HNMR (400 MHz, DMSO-d6) delta ppm 1.08-1.25 (m, 2H) 1.40 (s, 9H) 1.69-1.78 (m, 2H) 1.88-2.03 (m, 1H) 2.58-2.88 (m, 2H) 3.89-4.05 (m, 4H) 8.50-8.57 (m, 2H) |
94% | With potassium carbonate; In N,N-dimethyl-formamide; at 80℃; for 5h; | tert-Butyl 4-((methylsulfonyloxy)methyl)piperidin-l-carboxylate (Step 2 of Intermediate 1, 2.00 g, 6.82 mmol) was dissolved in DMF (80 mL). K2C03 (3.33 g, 10.23 mmol) was added thereto, followed by stirring for 5 minutes. 2-Chloropyrimidin-5-ol (890 mg, 6.82 mmol) was added thereto, followed by stirring at 80 C for 5h. To the reaction mixture, water was added, and the mixture was extracted with EtOAc. The organic layer was washed with saturated NH4C1 aqueous solution, dried with anhydrous MgS04, and then concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (EtOAc hexane = 30 % ~ 70 %) to obtain white solid (2.10 g, 94%). |
94% | t-butyl 4-((methylsulfonyloxy)methyl)piperidin-1-carboxylate (the product of synthesis step 2 of compound 431; 2.00 g, 6.82 mmol) was dissolved in DMF (80 mL). K2CO3 (3.33 g, 10.23 mmol) was added thereto, and stirred for 5 minutes. <strong>[4983-28-2]2-chloropyrimidin-5-ol</strong> (890 mg, 6.82 mmol) was added thereto, following with stirring at 80 C. for 5 hours. The reaction mixture was added with water, and extracted with EtOAc. The organic layer was washed with saturated NH4Cl aqueous solution, dried over anhydrous MgSO4, and concentrated under reduced pressure. The obtained concentrate was purified by silica gel column chromatography (EtOAchexane=30%?70%) to yield the title compound as white solid (2.10 g, 94%) |
With potassium carbonate; In N,N-dimethyl-formamide; at 80℃; | To a solution of 4-methanesulfonyloxymethylpiperidine-l-carboxylic acid tert-butyl ester (Preparation 23, 1.47g, 5.0mmol) and <strong>[4983-28-2]2-chloropyrimidin-5-ol</strong> (0.65g, 5.0mmol) in DMF (80mL) was added potassium carbonate (0.83g, 6.0mmol) and the reaction was heated to 80C until complete. The solvent was removed in vacuo, and the resulting residue was re- dissolved in EtOAc (300mL). The solution was washed with 1M NaOH solution (200mL), brine (200mL), then dried (MgS04) and the solvent was removed in vacuo. Purification by column chromatography (IH:EtOAc, 70:30) afforded the title compound: lH NMR delta?(400MHz, CDCI3): 8.30 (s, 1H), 4.18 (br. s., 2H), 3.92 (dd, J=6.25, 3.51 Hz, 2H), 2.78 (t, J=12.30 Hz, 2H), 2.09 - 1.95 (m, 1H), 1.84 (d, J=12.89 Hz, 2H), 1.49 (s, 9H), 1.41 - 1.24 (m,2H). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
70% | With potassium carbonate; In N,N-dimethyl-formamide; at 80℃; for 5.0h; | tert-butyl 4-((methylsulfonyloxy)methyl)piperidin-1-carboxylate ( 2.00 g, 6.81 mmol), 2-bromo-5- hydroxybenzonitrile (1.35 g, 6.87 mmol) and K2CO3 (1.88 g, 13.63 mmol) were dissolved in DMF (50 mL) at 80 C, following with stirring at the same temperature for 5 hours. The reaction mixture was added with water, and extracted with EtOAc. The organic layer was washed with saturated NH4Cl aqueous solution, dried over anhydrous MgSO4, and filtered. The filtrate was concentrated under reduced pressure. The concentrate was purified by column chromatography (Si02, 12 g cartridge; EtOAc / hexane = 0 % to 30 %), and concentrated to yield the title compound as white solid (1.90 g, 70%). |
70% | With potassium carbonate; In N,N-dimethyl-formamide; at 80℃; for 5.0h; | tert-Butyl 4-((methylsulfonyloxy)methyl)piperidin-l-carboxylate (Step 2 of Intermediate 1, 2.00 g, 6.81 mmol), <strong>[189680-06-6]2-bromo-5-hydroxybenzonitrile</strong> (1.35 g, 6.87 mmol) and K2C03 (1.88 g, 13.63 mmol) were dissolved in DMF (50 mL) at 80C. The solution was stirred at the same temperature for 5 h. To the reaction mixture, water was added, and the mixture was extracted with EtOAc. The organic layer was washed with saturated NH4C1 aqueous solution, dried with anhydrous MgS04, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (Si02, 12 g cartridge; EtOAc / hexane = 0 % to 30 %), and concentrated to obtain the desired compound (1.90 g, 70%) as white solid . |
70% | With potassium carbonate; In N,N-dimethyl-formamide; at 80℃; for 5.0h; | t-butyl 4-((methylsulfonyloxy)methyl)piperidin-1-carboxylate (2.00 g, 6.81 mmol), <strong>[189680-06-6]2-bromo-5-hydroxybenzonitrile</strong> (1.35 g, 6.87 mmol) and K2CO3 (1.88 g, 13.63 mmol) were dissolved in DMF (50 mL) at 80 C., following with stirring at the same temperature for 5 hours. The reaction mixture was added with water, and extracted with EtOAc. The organic layer was washed with saturated NH4Cl aqueous solution, dried over anhydrous MgSO4, and filtered. The filtrate was concentrated under reduced pressure. The concentrate was purified by column chromatography (SiO2, 12 g cartridge; EtOAchexane=0% to 30%), and concentrated to yield the title compound as white solid (1.90 g, 70%) |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
59 g | With caesium carbonate; In N,N-dimethyl-formamide; at 80℃; for 2h; | To a mixture of tert-butyl 4-[(methylsulfonyl)oxy]methyl}piperidine-1 -carboxylate (I-48) (54.6 g, 186 mmol) and Cs2C03 (1 10 g, 339 mmol) in DMF (400 mL) was added <strong>[39903-01-0]2-amino-5-bromopyridin-3-ol</strong> (I-53) (32.0 g, 169.3 mol) at room temperature. The resulting mixture was stirred at 80 °C for 2 h. TLC (petroleum ether/ EtOAc = 1 /1) showed the reaction was complete. The reaction mixture was cooled to room temperature and concentrated under high vacuum to dryness. The residue was diluted with 200 mL of water and extracted with CH2CI2 (3 x 200 mL). The combined organic layers were washed with brine (100 mL), dried over Na2S04. The mixture was concentrated under vacuum to give the crude product, which was purified by silica gel chromatography (petroleum ether/ EtOAc = 3/1 to 1 /2) to give tert-butyl 4-[(2-amino-5-bromopyridin-3-yl)oxy]methyl}piperidine-1 -carboxylate (I-54) (59.0 g, 90percent) as a white solid. |
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