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Design and Development of Antibiotics and Synthesis of Modified Nucleosides
Hegde, Pooja Venkatesh ;
Abstract: Tuberculosis (TB) remains a leading cause of infectious disease mortality and morbidity resulting in nearly 1.3 million deaths annually and infecting nearly one-quarter of the population. Chapter 1 discusses the epidemiology, control, and management of TB. Isoniazid (INH) is a cornerstone for the treatment of drug-susceptible TB, yet the quantitative structure-activity relationships for INH are not well documented in the literature. Chapter 2 evaluates a systematic series of INH analogs against contemporary Mycobacterium tuberculosis (Mtb) strains from different lineages and several key species of non-tuberculous mycobacteria (NTM). To assess the specific activity of this series of INH analogs against mycobacteria, we assayed them against a panel of Gram-positive and Gram-negative bacteria, as well as a number of fungi. Our findings provide an updated analysis of the structure-activity relationship of INH that will serve as a valuable resource for the development of a next generation antitubercular compounds. para-Aminosalicylic acid (PAS), is an important second-line agent for treating drug-resistant Mtb. PAS has a moderate bioavailability and rapid clearance that necessitate high doses in order to facilitate an effective treatment. Consequently, such high doses commonly results in gastrointestinal disturbances (presumably by disruption of gut microbiota and host epithelial cells). Chapter 3 discusses the design, synthesis and evaluation of PAS prodrugs and analogs with improved oral bioavailability, thereby preventing intestinal accumulation as well as undesirable bioactivation by the gut microbiome to cytotoxic folate species. The pivoxyl prodrug and fluorination at the 5- position address the primary limitations of PAS and have the potential to revitalize this second-line TB drug. vi Pyrazinamide (PZA) is a critical component of the first-line TB treatment regimen because of its sterilizing activity against non-replicating Mtb, but its mechanism of action has remained enigmatic. PZA is a prodrug converted to the active moiety- pyrazinoic acid (POA), by pyrazinamidase, an amidase within the nicotinamide adenine dinucleotide (NAD) salvage pathway encoded by pncA in Mtb. PZA resistance is most commonly induced via loss-of-function mutations within PncA. It has recently been demonstrated that POA induces targeted protein degradation of the enzyme PanD, a crucial component of the coenzyme A biosynthetic pathway, essential in Mtb. Chapter 4 describes the structure activity relationship (SAR) evaluation of various POA analogs. Further development and mechanistic analysis of these analogs may lead to a next generation POA analog for treating TB. The salicylic acid derived small molecule siderophores known as mycobactins are essential for mycobacterial iron acquisition and mycobactin biosynthesis has been biochemically and genetically validated as essential for survival in vivo. Sal-AMS, a modified nucleoside derivative that mimics an intermediate in the mycobactin biosynthetic pathway, is a potent Mtb inhibitor. Chapter 5 explores polyfluorinated salicylic acid derivatives as antimetabolites, designed to antagonize mycobactin synthesis. Enzymatic studies demonstrated that the tri- and tetra- fluorinated salicylic acid analogs were neither substrates nor inhibitors of MbtA, but the di-fluorinated compounds were readily activated by the bifunctional adenylating enzyme MbtA, responsible for processing salicylic acid moieties for synthesis of mycobactins. However further microbiological analysis revealed that the polyfluorinated derivatives have low potential as anti-TB agents and do not appear to operate by inhibiting mycobactin synthesis. vii Lastly, the chemical synthesis of nucleoside analogs, an important class of compounds with applications as anti-infective, anti-cancer, and diagnostic agents, is extremely challenging, typically requiring linear synthesis, multiple protectiondeprotection sequences, and stereoselective formation of the critical glycosidic linkage. Nucleoside phosphorylases (NPs) have tremendous biocatalytic potential and aid in the selective modification of nucleosides under green conditions. Chapter 6 focuses on the functional characterization of 15 thermostable purine NPs. Four of these were further selected for a comprehensive analysis of their substrate scope. Next, chemoenzymatic methods for the synthesis of modified nucleosides were extensively studied using various coupled systems, and 12 modified nucleosides were synthesized, isolated, and characterized. viii Table of Contents Acknowledgments......................
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Reinvestigation of the structure-activity relationships of isoniazid
Hegde, Pooja ; Boshoff, Helena I. M. ; Rusman, Yudi ; Aragaw, Wassihun Wedajo ; Salomon, Christine E. ; Dick, Thomas , et al.
Abstract: Isoniazid (INH) remains a cornerstone for treatment of drug susceptible tuberculosis (TB), yet the quant. structure-activity relationships for INH are not well documented in the literature. In this paper, we have evaluated a systematic series of INH analogs against contemporary Mycobacterium tuberculosis strains from different lineages and a few non-tuberculous mycobacteria (NTM). Deletion of the pyridyl nitrogen atom, isomerization of the pyridine nitrogen to other positions, replacement of the pyridine ring with isosteric heterocycles, and modification of the hydrazide moiety of INH abolishes antitubercular activity. Similarly, substitution of the pyridine ring at the 3-position is not tolerated while substitution at the 2-position is permitted with 2-methyl-INH 9 displaying antimycobacterial activity comparable to INH. To assess the specific activity of this series of INH analogs against mycobacteria, we assayed them against a panel of gram-pos. and gram-neg. bacteria, as well as a few fungi. As expected INH and its analogs display a narrow spectrum of activity and are inactive against all non-mycobacterial strains evaluated, except for 4, which has modest inhibitory activity against Cryptococcus neoformans. Our findings provide an updated anal. of the structure-activity relationship of INH that we hope will serve as useful resource for the community.
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Keywords: Isoniazid ; SAR
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CAS No. : | 613-94-5 |
Formula : | C7H8N2O |
M.W : | 136.15 |
SMILES Code : | O=C(NN)C1=CC=CC=C1 |
MDL No. : | MFCD00007596 |
InChI Key : | WARCRYXKINZHGQ-UHFFFAOYSA-N |
Pubchem ID : | 11955 |
GHS Pictogram: |
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Signal Word: | Warning |
Hazard Statements: | H302-H315-H319-H335-H351 |
Precautionary Statements: | P201-P202-P261-P264-P270-P271-P280-P301+P312-P302+P352-P304+P340-P305+P351+P338-P308+P313-P330-P332+P313-P362+P364-P403+P233-P501 |
* 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 |
---|---|---|
Step 1: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-(5-phenyl-[1,3,4]oxadiazol-2-yl)-amine 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (1 g, 3.6 mmol), triethylamine (1 mL, 7.2 mmol) and diphenylphosphoryl azide (1.98 g, 7.2 mmol) were placed in toluene (20 mL) and heated to 60 C. for 20 minutes. Benzoic acid hydrazide (0.98 g, 7.2 mmol) was added in a single portion and the reaction was heated and stirred for 2 hours. The reaction was cooled and submitted to aqueous workup. The crude material was dissolved in dioxane (100 mL), phosphorous(V) oxychloride was added and the reaction was heated to 100 C. overnight. The reaction was cooled and worked up and the residue was purified on silica gel (0-80% EtOAc in hexanes) to yield the title compound. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
91% | With sulfuric acid; acetic acid; In methanol; water; for 1h;Reflux; | General procedure: Compounds of series I were synthesized by refluxing <strong>[92-55-7]5-nitro-2-furaldehyde diacetate</strong> 98% (5 mmol) and benzhydrazides (3) (5 mmol) in water, sulphuric acid, acetic acid, and methanol (8:7:8:20 v/v) for 1 h. After cooling, the mixture was poured into cold water to precipitate the azomethine derivatives 20(see structural elucidation of the compounds of series I in Supplementary data, p. S2). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
91.5% | With acetic acid; In ethanol; at 70℃; for 5h; | General procedure: The Compound I (0.79 mmol)was dissolved in 10 mL of absolute ethanol. Then the mixture of R2-CHO (0.66 mmol), 10 mL acetic acid and 3 mL absolute ethanol was added dropwise at 70 °C. The mixture was refluxed for 5 h. The solvent was evaporated. The crude product was purified by flash chromatography (neutral Al2O3). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
76% | With acetic acid; In acetonitrile; for 9.0h;Reflux; | General procedure: A mixture of benzoylhydrazine (1.22 g, 0.01mol), dimethylformamide-dimethylacetal (1.91 g,0.01 mol)) and sulfa-drugs (0.012 mol) in dry acetonitrile(15 mL) containing acetic acid (3 mL) was refluxed for 9 h., then left to cool. The solid product formed was collected by filtration and recrystallized from dioxane to give compounds 1-13, respectively. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
42% | General procedure: Trifluoromethyl ketones (1; 0.30 mmol, 1 equiv), acylhydrazines (2; 0.45 mmol, 1.5 equiv) and TsOH (20 mol%) were added to a dried round-bottom flask (50 mL) fitted with a magnetic bar. 1,4-Dioxane 4 mL) was then added and the mixture was stirred and heated to reflux. After the formation of acylhydrazones (monitored by TLC), tin powder (1.35 mmol, 4.5 equiv) and ethyl 2-(bromomethyl)acrylate (3; 1.20 mmol, 4 equiv) were added to the flask. When acylhydrazones had essentially disappeared (monitored by TLC), the reaction mixture was cooled to r.t., then 1,4-dioxane was removed under vacuum. Saturated NH4Cl solution (10 mL) was added and the mixture was stirred for 10 min. The mixture was extracted with EtOAc (3 × 10 mL) and the combined organic phases were dried (MgSO4) and concentrated. Purification of the residue by silica gel column chromatography (petroleum ether-EtOAc, 4:1) furnished the pure products 4. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
18% | In isopropyl alcohol; at 90℃; | General procedure: A mixture of 2-phenylacetohydrazide (1) (0.10?g, 0.67?mmol) and 1,1,1-trifluoro-5-phenylpentane-2,4-dione (3a) (0.14?g, 0.67?mmol) in a solution of i-PrOH (5?mL) was heated at 90?C for 48?h. After cooling to room temperature, EtOAc and water were added. The EtOAc extract was washed with water, brine and dried (Na2SO4). Flash chromatography (petroleum ether/EtOAc; 100:0 to 93:7) followed by recrystallization from Et2O/petroleum ether gave 4 (0.17?g, 71%), mp 122-123?C (Et2O/petroleum ether). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
10% | In isopropyl alcohol; at 90℃; | General procedure: A mixture of 2-phenylacetohydrazide (1) (0.10?g, 0.67?mmol) and 1,1,1-trifluoro-5-phenylpentane-2,4-dione (3a) (0.14?g, 0.67?mmol) in a solution of i-PrOH (5?mL) was heated at 90?C for 48?h. After cooling to room temperature, EtOAc and water were added. The EtOAc extract was washed with water, brine and dried (Na2SO4). Flash chromatography (petroleum ether/EtOAc; 100:0 to 93:7) followed by recrystallization from Et2O/petroleum ether gave 4 (0.17?g, 71%), mp 122-123?C (Et2O/petroleum ether). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
37% | In isopropyl alcohol; at 90℃; | General procedure: A mixture of 2-phenylacetohydrazide (1) (0.10?g, 0.67?mmol) and 1,1,1-trifluoro-5-phenylpentane-2,4-dione (3a) (0.14?g, 0.67?mmol) in a solution of i-PrOH (5?mL) was heated at 90?C for 48?h. After cooling to room temperature, EtOAc and water were added. The EtOAc extract was washed with water, brine and dried (Na2SO4). Flash chromatography (petroleum ether/EtOAc; 100:0 to 93:7) followed by recrystallization from Et2O/petroleum ether gave 4 (0.17?g, 71%), mp 122-123?C (Et2O/petroleum ether). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
18% | In isopropyl alcohol; at 90℃; | General procedure: A mixture of 2-phenylacetohydrazide (1) (0.10?g, 0.67?mmol) and 1,1,1-trifluoro-5-phenylpentane-2,4-dione (3a) (0.14?g, 0.67?mmol) in a solution of i-PrOH (5?mL) was heated at 90?C for 48?h. After cooling to room temperature, EtOAc and water were added. The EtOAc extract was washed with water, brine and dried (Na2SO4). Flash chromatography (petroleum ether/EtOAc; 100:0 to 93:7) followed by recrystallization from Et2O/petroleum ether gave 4 (0.17?g, 71%), mp 122-123?C (Et2O/petroleum ether). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
83% | With potassium carbonate; dimethyl sulfate; In water; at 50℃; for 16h;Green chemistry; | General procedure: A suspension of thiourea (1) (0.076 g, 1 mmol), hydrazides 3a-j (1 mmol), dimethyl sulfate (2) (0.063 g, 0.5 mmol) and potassium carbonate (0.069 g, 0.5 mmol) in 4 mL water was heated at 50 C for 15-20 h. The reaction progress was checked with TLC. The mixture was cooled in an ice bath in order to increase in total precipitation. The solid was fltered of, washed with cold water and ethanol, and dried in an oven at 60 C without need for further purifcation. 3(5)-Phenyl-1H-1,2,4-triazol-5(3)-amine (4a): White solid; FTIR (KBr) nu: 3415, 2981, 2364, 1643, 1551, 1364, 1290, 1142, 694 cm-1; 1H NMR (400 MHz, DMSO-d6) delta: 11.86 (1H, br, NH), 7.91-7.78 (2H, m, H-2,6), 7.53-7.25 (3H, m, H-3,4,5), 6.80 (2H, br, NH2) ppm; 13C NMR (100 MHz, DMSO-d6) delta: 161.6, 163.6 (C-3, C-5), 132.2 (C-1), 128.4 (C-3,5), 128.0 (C-4), 127.1 (C-2,6) ppm. Anal. Calcd for C8H8N4: C 59.99, H 5.03, N 34.98; found: C 60.02, H 5.04, N 34.94. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
97% | With C83H64AuF6N3O6S2; In chlorobenzene; at 60℃; for 42h;Schlenk technique; | General procedure: In a small Schlenk flask equipped with a small stirring bar the corresponding alkyne (0.5 mmol) and the corresponding benzhydrazide (0.5 mmol) were mixed with anisole or chlorobenzene (2 mL). For the internal standard mesitylene (69 μL, 0.5 mmol) was added. A stock solution of the corresponding gold-triflimide catalyst (0.005 M in anisole or chlorobenzene, 200 μL, 0.001 mmol, 0.2 mol %) was introduced to the mixture. The reaction mixture was stirred at 60 C or 80 C for the respective time and the progress of the reaction was monitored by GC. After the reaction was completed, the solvent was removed in vacuo and the residue triturated in pentane. Column chromatography (DCM/ethyl acetate 5:1) afforded the respective benzohydrazone in good to excellent yields. |