Method Development and Validation of Gallic Acid in Liquid Dosage Form by Using RP-HPLC Method.

A simple, rapid, precise, sensitive, and reproducible reverse-phase high-performance liquid chromatography (RP-HPLC) method has been developed for the quantitative analysis of gallic acid in the pharmaceutical dosage form. Chromatographic separation of gallic acid was achieved on Waters Alliance-e 2695, by using Waters X-Terra RP-18 (150 × 4.6 mm, 3.5 µ) column and the mobile phase containing 0.1% formic acid and ACN in the ratio of 70:30% v/v. The flow rate was 1.0 mL/min; detection was carried out by absorption at 275 nm using a photodiode array detector at ambient temperature. The number of theoretical plates and tailing factor for gallic acid was NLT 2000 and should not be more than 2 respectively. Percentage relative standard deviation of peak areas of all measurements is always less than 2.0. The proposed method was validated according to ICH guidelines. The method was found to be simple, economical, suitable, precise, accurate, and robust method for quantitative analysis of gallic acid.

Screening of Phyllanthus niruri Leaves Phytoconstituents for Antiviral and Antibacterial Activity by Molecular Docking Studies.

The genus Phyllanthus belongs to one of the largest plant families, the Phyllantaceae (L.). Phyllanthus niruri is an annual perennial herb that grows in tropical Asia, America, China, and the islands of the Indian Ocean. Numerous alkaloids, steroids, flavonoids, lignans, coumarins, polyphenols, and lipids are present in Phyllanthus. The effects of plants have been studied for a variety of purposes, including their antioxidant (Giribabu et al., Evid Based Complement Alternat Med, 2014), anti-inflammatory (Porto et al., Revista Brasileira de Pharmacognosy, 2013), antinociceptive (Sathisha et al., Indian Drugs, 2009), analgesic (Mostofa et al., BMC Complement Altern Med, 2017), antiulcer (Mali et al., Biomed Aging Pathol, 2011), antiarthritic (Obidike and Salawu, Planta Medica, 2010), antiplasmodial (Shilpa et al., Environ Dis, 2018), immunomodulatory (Manikkoth et al., Anticonvulsant activity of Phyllanthus amarus in experimental animal models), anticonvulsant (Wasnik et al., Int J Pharm Sci Rev Res, 2014), antidepressant (Venkateswaran et al., Effects of an extract from Phyllanthus niruri on hepatitis B and woodchuck hepatitis viruses: In vitro and in vivo studies (antiviral agent/Marmota monax/DNA polymerase/hepatitis B surface antigen/woodchuck hepatitis surface antigen). In Hepatitis B and The Prevention of Primary Cancer of The Liver: Selected Publications of Baruch S Blumberg, pp 535-539), antiviral (Venkateswaran et al., Effects of an extract from Phyllanthus niruri on hepatitis B and woodchuck hepatitis viruses: In vitro and in vivo studies (antiviral agent/Marmota monax/DNA polymerase/hepatitis B surface antigen/woodchuck hepatitis surface antigen). In Hepatitis B and The Prevention of Primary Cancer of The Liver: Selected Publications of Baruch S Blumberg, pp 535-539), antitumor (Sharma et al., Asian Pac J Cancer Prev, 2009), hyperlipidemia (Khanna et al., J Ethnopharmacol, 2002), and antifertility (Ezeonwu, Inquiries J, 2011). For additional docking investigations with distinct proteins, the leaf chemicals are assessed, that is, the crystal structure of serine protease hepsin in complex with inhibitor [PDB ID:5 CE1] for antiviral activity human topoisomerase II beta in complex with DNA and etoposide [PDB ID:3QX3] and crystal structure of E. coli GyraseB 24 kDa in complex with 4-(4-bromo-1H-pyrazol-1-yl)-6-[(ethylcarbamoyl)amino]-N-(pyridin-3-yl) pyridine-3-carboxamide [PDB ID: 6F86] for antibacterial activity and have been selected. To evaluate the in silico results and grading of virtual screening, or molecular docking, ritonavir antiviral activity and ampicillin for antibacterial activity were used as a benchmark.

Screening of Phyllanthus niruri Plant Active Constituents for Anticancer and Antifungal Activity by Insilico Methods.

A large genus of shrubs, trees, and rare plants belonging to the Euphorbiaceae family, Phyllanthus contains 600-700 species. The Phyllanthus niruri (L.) species is a tiny, erect annual herb that can reach heights of 30-40 cm. Its 7-12 cm long, sessile, alternating leaves are native to the Amazon rainforest, but they can also be found in other tropical regions such as South East Asia, Southern India, America, China, and the islands of the Indian Ocean. Phyllanthus contains many classes of alkaloids, steroids, flavonoids, lignin, polyphenols, and lipids. Numerous activities of the plant have been studied, including antidepressant (Wasnik et al., Int J Pharm Sci Rev Res, 6:26-29, 2014), anticancer (Sayuti et al. Studies, 10:17, 2020), anti-inflammatory, antinociceptive (Porto et al., Revista Brasileira de Farmacognosia, 23:138-144, 2013), analgesic (Bhat et al., Pharm Res, 7:378, 2015), antiarthritic (Mali et al., Biomed Aging Pathol, 1:185-190, 2011), immunomodulatory, antibacterial, antifungal (Shilpa et al., Environm Dis, 3:63, 2018), antidiabetic (Kumar et al., Biomed Pharm J, 12:57-63, 2019), antiulcer (Mostofa et al., BMC Complement Altern Med, 17:1-10, 2017), antiviral (Wahyuni et al., Malays Appl Biol, 48:105-111, 2019), antiplasmodial (Ifeoma et al., Asian Pacific J Trop Med, 6:169-175, 2013), anticonvulsant (Amaechina and Omogbai, Nig J Nat Prod Med, 17:61-65, 2013), and hepato human cytochrome P450 CYP17A1 in association with abiraterone [PDB ID: 3RUK] plant extracts. New selective androgen receptor modulators were synthesized, and they were biologically evaluated (SARMs) (Micah et al., J Veter Med Anim Health 5(1):8-15, 2013, Rusmana et al., Indonesian Biomed J 9(2):84-90, 2017, Al Zarzour et al., Nutrients 10(8):1057, 2018, Khanna J Ethnopharmacol 82(1):19-22, 2002). In the present study [PDB ID: 3RUK,5T8E] with anticancer and [PDB ID: 6F0E,1EA1] with antifungal activities were used for docking study. In this study fluconazole's antifungal activity and dacarbazine's anticancer activity were used as benchmarks for molecular docking with Schrodinger 13.0 to compare the activity of Phyllanthus niruri's active constituents.

Screening of Phyllanthus niruri Root Phytoconstituents for Antibacterial, Antifungal, Anticancer, and Antiviral Activities by Molecular Docking Studies.

The systematic exploitation of the structural variety of natural products is made possible by docking studies, which have been shown to be a crucial technique. This study's goal was to evaluate various activities for the chemicals in the root portion of Phyllanthus niruri. This plant's constituents are active in a variety of ways. In order to develop drugs, molecules with such a framework have been utilized as the lead. Schrodinger Maestro (v13.0) software was used to conduct a molecular docking analysis of root components with certain proteins linked to the illnesses. In comparison to commercially available conventional medications, molecular docking data also demonstrated greater scores. For additional docking investigations with distinct proteins, the root chemicals are assessed, that is, crystal structure of serine protease hepsin in complex with inhibitor [PDB ID:5 CE1] for antiviral activity, human topoisomerase II beta in complex with DNA and etoposide [PDB ID:3QX3], and crystal structure of E. coli GyraseB 24 kDa in complex with 4-(4-bromo-1H-pyrazol-1-yl)-6-[(ethylcarbamoyl)amino]-N-(pyridin-3-yl) pyridine-3-carboxamide [PDB ID: 6F86] for antibacterial activity, Cytochrome P450 14 alpha-sterol demethylase (CYP51) from Mycobacterium tuberculosis in complex with fluconazole [PDB ID:1EA1], and structure of yeast Sec14p with a picolinamide compound [PDB ID:6F0E] for antifungal activity and synthesis and biological evaluation of novel selective androgen receptor modulators (SARMs). Part II: Optimization of 4-(pyrrolidin-1-yl) benzonitrile derivatives [PDB ID: 5T8E] and Human Cytochrome P450 CYP17A1 in complex with Abiraterone [PD B ID:3RUK] for anticancer activity have been selected. Ritonavir's antiviral activity, ampicillin's ability to treat bacterial infections, fluconazole's ability to treat fungi, and dacarbazine's ability to treat cancer were utilized as benchmarks to assess the in silico outcomes and grading of virtual screening or molecular docking.

Screening of Spirulina Components for Anti-Parkinson's and Anti-Alzheimer's Activity by in Silico Methods and Docking Studies.

Spirulina platensis was first isolated from Lake Texcoco by Aztecs in the sixteenth century. In 2012, spirulina was considered to be safe dietary supplement by the Food and Drug Administration (FDA). Spirulina is a cyanophytic microalgae that is often considered as single cell protein. It contains many essential amino acids, proteins, fatty acids, antioxidant pigments, carotenoids, and cyanogenic pigments, that is, phycocyanobilins and phycocyanins (Eriksen, Appl Microbiol Biotechnol, 80(1):1-4, 2008). Components of spirulina are investigated for many health benefits and for pharmaceutical uses (Karkos et al., Spirulina in clinical practice: evidence-based human applications). Spirulina has been found to have a role in growth, immunity (Wu et al., Arch Toxicol, 90(8):1817-40, 2016), antioxidant (Wu et al., Arch Toxicol, 90(8):1817-40, 2016), antiviral (Ayehunie et al., J Acquir Immune Defic Syndr Hum Retrovirol, 18(1):7-12, 1998), antitoxicologic, anti-cancerogenic (Hirahashi et al., Int Immunopharmacol, 2(4):423-34, 2002), antidiabetic (Layam and Reddy, Diabetol Croat, 35(2):29-33, 2006), and neuroprotective properties. In this study, we focused on spirulina components in anti-Parkinson's and anti-Alzheimer's activity. Four potential targets, two for each activity, that is, structure of parkinE3 ligase (PDB ID:4I1H) and structure of BACE bound to 5-(3-(5-chloropyridin-3-yl)phenyl)-5-cyclopropyl-2-imino-3-methylimidazolidin-4one (PDBI D:4DJx) for anti-Parkinson's activity and structure of human MAO B in complex with selective inhibitor safinamide (PDB ID:2V5Z) and crystal structure of human BACE-1 in complex with CNP520(PDB ID:6EQM) for anti-Alzheimer's activity, have been selected. The in silico results and scoring of virtual screening, that is, molecular docking, were compared with commonly used marketed drugs such as levodopa for Parkinson's disease (PD) and rivastigmine (Rösler et al., BMJ, 318(7184):633-40, 1999) for Alzheimer's disease.

Molecular Docking Studies of Phyllanthus niruri Root Phytoconstituents for Antibreast Cancer Activity Using Multiple Proteins.

The systematic exploitation of the structural variety of natural products is made possible by docking studies, which have been shown to be a crucial technique. The objective of the current work was to use molecular docking studies with different proteins to identify acceptable and efficient compounds from root phytoconstituents of Phyllanthus niruri plant. Numerous human disorders have been treated using Phyllanthus niruri. Antioxidant, antibacterial, antifungal, anti-inflammatory, antipyretic, antimalarial, antiviral, immunomodulatory, antidepressant, anticonvulsant, antinociceptive, anti-ulcer, anti-arthritic, anticancer, hyperlipidemia, and antifertility were only a few of its many pharmacological effects. One of the most prevalent malignancies in women worldwide, breast cancer is one of the leading causes of mortality worldwide. The most successful breast cancer treatments now on the market have negative side effects and are useless in people. In order to develop drugs, molecules with such a framework have been utilized as the lead. Schrodinger Maestro (v13.0) software was used to conduct a molecular docking analysis of root components with certain proteins linked to the illnesses. In comparison to commercially available conventional medications, molecular docking data also demonstrated greater scores. Dacarbazine's ability to treat cancer was utilized as benchmark to assess the in silico outcomes and grading of virtual screening or molecular docking.

Repurposing Marketed Drugs as Antidepressants with In Silico, In Vivo, and In Vitro Screening Methods.

Drug repurposing is a strategy to identify new therapeutic uses for marketed drugs, discontinued or shelved drugs, and drug candidates in clinical development. Drug repurposing has gained momentum over the past few years. A slow rate of new drug discovery and higher cost of new drug development attracted the attention for repurposing and repositioning of old medications . A deeper understanding of the pathogenesis of depression encourages novel discoveries through drug repurposing to treat depression. In this study, indole- and isatin-containing drugs are going to be repurposed using computational methods for the treatment of depression.

Impurity Profiling and Identification of 2,6-Diisopropylphenol by HPLC Method and Raman Spectroscopy Method.

Identification of 2,6-diisopropylphenol by Raman spectroscopy using the TruScan Raman spectrum is validated by specificity test and robustness. HPLC assay method is developed to detect the 2,6-diisopropylphenol and its impurities A = determination of 2,6-diisopropylphenol; its main impurities related compound A = 2,6-diisopropylphenyl isopropyl ether, related compound B = 2,6-diisopropylquinone, and related compound C = 3,3'-5,5'-tetraisopropyl diphenol; and unknown impurities done by HPLC method. Related compound A and C determination has been developed by normal phase HPLC; good resolution peak shapes have resulted from the peak results. The limit of impurities A and C is reported to be 0.05% and 0.03%, and the limit of impurity B is also detected by using the same above procedure, but the column has been changed for the maximum wavelength detection and better elution from the peak results. The reported impurity level was 0.02%, the unknown impurity limit was 0.0149%, and the total impurity level of 2,6-diisopropylphenol was reported to be 0.06% which are in the threshold limit level. It specifies that the drug is safe and efficient without any toxicity.

Design, Characterization, and Docking Studies of Some Novel Isatin Derivatives for Anticonvulsant and Antidepressant Activity.

Isatin (indoline-2,3-dione) derivatives are derived from plant origin indole derivatives by the Sandmeyer method and characterized by IR, NMR, and mass spectrometric method. Molinspiration is used to calculate the molecular properties of all the synthesized compounds and to generate bioactivity scores (GPCR ligand, ion channel inhibitor, kinase inhibitor, nuclear receptor ligand, protease inhibitor, enzyme inhibitor) of the series of compounds. ADME predicted parameters are lipophilicity, P-gp substrate, GI absorption, bioavailability, lead-likeness, and blood-brain barrier (BBB) permeability by boiled egg model. Molecular docking is performed for the synthesized compounds they compared with the standard drugs.

Synthesis, Molecular Docking Studies and Biological Evaluation of N-Acylarylhydrazones as Anti-Inflammatory Agents.

In the present work a series of N'-arylidene-2-(benzamido)-3-(naphthalen-2-yl)acrylohydrazides were synthesized by refluxing the intermediate 2-(benzamido)-3-(naphthalen-2-yl)acrylohydrazide with various substituted benzaldehyde in the presence of glacial acetic acid. The intermediate 2-(benzamido)-3-(naphthalen-2-yl)acrylohydrazide 2 was prepared by stirring 4-((naphthalen-2-yl)methylene)-2-phenyloxazol-5(4H)-one with hydrazine hydrate in the presence of absolute ethanol. The chemical structures of the compounds were established by IR, 1H NMR and mass spectral data. All the compounds were evaluated for anti-inflammatory (in vivo, in vitro) activity and performed docking against COX-2. The compounds 3a, 3c and 3o showed good inhibition of COX-2 in in vitro studies (0.75 µM, 0.5 µM and 0.7 µM as IC50, respectively). The compounds 3c, 3e and 3f were found to be more active than standard drug phenylbutazone at equidose. Molecular docking studies showed that compound 3 m exhibited good binding affinity against COX-2 with docking score 9.328 kcal/mol, when compared to the standard celecoxib.

Synthesis and Characterization of Biologically Significant 5-[N,N-dialkylamino alkoxy] azaindole 2-one, 3-thiosemicarbazones and 5-[N,N-dialkylamino alkoxy] azaindole 3-hydrazone, 2-ones.

In the present work, new indole derivatives, i.e., 5-[N,N-di alkyl amino alkoxy] azaindole 2,3- di-one derivatives, are synthesized and characterized. These compounds were subjected to acute toxicity and then screened for antiepileptic activity on maximal electroshock seizure (MES) model in albino Wistar rats. In that study 5-[2-dimethyl amino ethoxy] Azaindole-3-hydrazone,2-one and 5-[2- dimethyl amino ethoxy] Azaindole 2-one,3-thiothiosemicarbazone(IIIa) showed good antiepileptic activity and less neurotoxicity compared to phenytoin. The purpose of the present study is to investigate the effect of 5-[2-dimethyl amino ethoxy] Indole 2,3- di one and 5-[2-dimethyl amino ethoxy] Azaindole 2-one,3-thiosemicarbazone(IIIa) derivatives on biogenic amines concentrations in rat brain after induction of seizures by MES method. The aim of study was relationship between seizure activities and altered the monoamines such as Noradrenaline (NA), Dopamine (DA), Serotonin (5-HT) in forebrain of rats in MES seizure models. In MES model, study of 5-[2-dimethyl amino ethoxy] Azaindole 3-hydrazone,2-one(Va) and 5-[2-dimethyl amino ethoxy]Azaindole 2-one,3-thiosemicarbazone(IIIa) (100 mg/kg) showed significant restoration of the decreased levels of brain monoamines such as noradrenaline, dopamine, and 5-hydroxytryptamine. Thus, this study suggests that 5-[2-Dimethyl amino ethoxy] Azaindole 3-hydrazone,2-one (V) and 5-[2-dimethyl amino ethoxy] Azaindole 2-one,3-thiosemicarbazone (IIIa) increased the monoamines on rat brain, which may decrease the susceptibility to MES-induced seizure in rats.

Evaluation of Anti-Epileptic Effect of New Indole Derivatives by Estimation of Biogenic Amines Concentrations in Rat Brain.

The new heterocyclic compounds are used to treat epilepsy. In the present work new indole derivatives i.e. 5-[2(3)-di alkyl amino alkoxy] Indole 2,3-di-one derivatives are synthesized and characterized and these compounds was subjected to acute toxicity and then screened for antiepileptic activity on Maximal Electroshock (MES) seizures model in albino wistar rats. In that study 5-[2-dimethyl amino ethoxy] Indole 2,3 dione and 5-[2-dimethyl amino ethoxy] Indole 2-one,3-semicarbazone(IVa) showed good antiepileptic activity and less neurotoxicity compared to phenytoin. The purpose of the present study is to investigate the effect of 5-[2-dimethyl amino ethoxy] Indole 2,3-di one and 5-[2-dimethyl amino ethoxy] Indole 2-one,3-semicarbazone(IVa) derivatives on biogenic amines concentrations in rat brain after induction of seizures by Maximal Electro Shock(MES) method. Our aim of study was relationship between seizure activities and altered the monoamines such as noradrenaline (NA), dopamine (DA), serotonin (5-HT) in forebrain of rats in MES seizure models. In MES model, study of 5-[2-dimethyl amino ethoxy] Indole 2,3 dione(IIIa) and 5-[2-dimethyl amino ethoxy] Indole 2-one,3-semicarbazone(IVa) (100 mg/kg) showed significantly restored the decreased levels of brain monoamines such as Noradrenaline, Dopamine & 5-Hydroxy Triptamine. Thus, this study suggests that study of 5-[2-Dimethyl amino ethoxy] Indole 2,3-dione(IIIa) and 5-[2-dimethyl amino ethoxy] Indole 2-one,3-semicarbazone(IVa) increased the monoamines on rat brain, which may be decreased the susceptibility to MES induced seizure in rats.

Development of Novel Indole Molecules for the Screening of Anti-Inflammatory Activity.

In the present work, some new 5-[2(3)-dialkylamino alkoxy] Indole 2,3-diones were prepared from 5-hydroxy isatin. A mixture of 5-hydroxy isatin, dialkylamino alkylhalide in alcoholic potassium hydroxide was stirred at room temperature for 6 h to get the 5-[2(3)-dialkylamino alkoxy] Indole 2,3-diones. The structures of the products were characterized by IR, NMR, MASS Spectral studies. All the compounds were evaluated for anti-inflammatory activity. Some of these compounds showed good anti-inflammatory activity compared with standard compound Indomethacin.

Synthesis and screening of biologically significant 5-hydroxy isatin derivatives for antioxidant activity.

In the present work, some new 5-Hydroxyindole 3-thiosemicarbazone 2-ones and 5-[2(3)-dialkylaminoalkoxy] Indole 3-hydrazone 2-ones were prepared from 5-hydroxy isatin. The structures of the products were characterized by IR, NMR, and MASS Spectral studies. All the compounds were examined for antioxidant activity by using 1,1-diphenyl-2-picryl-hydrazyl and hydrogen peroxide (H2O2), and the total antioxidant capacity by a phosphomolybdenum assay. In general, the derivatives were found to exhibit antioxidant activity. Further, the compounds with dialkyl amino alkoxy at the C5 position demonstrated significant antioxidant activity.

Synthesis and antiepileptic activity of schiff's bases of dialkylamino alkoxy isatin derivatives.

In the present work, some new 5-[2(3)-dialkylamino alkoxy] Indole 3-thiosemicarbazone 2-ones and 5-[2(3)-dialkylamino alkoxy] Indole 3-hydrazone 2-one were prepared from 5-hydroxy isatin. The structures of the products were characterized by IR, NMR, and MASS Spectral studies. All the compounds were examined for antiepileptic activity by maximal electroshock seizure (MES) and pentylenetetrazole (PTZ) induced convulsion method. These compounds were also evaluated for their neurotoxicity study by rotarod method. Some of these compounds showed good antiepileptic activity when compared with standard drug Phenytoin and all the compounds showed less neurotoxicity when compared with standard drug Diazepam.