Challenges and Advances in Pharmaceutical Research Vol. 2

The rise in pathogen resistance is a serious concern for health care in the twenty-first century, and effective vaccines to prevent life-threatening diseases are urgently needed. In order to uncover new antigens with diagnostic and vaccine potential, pathogens have been studied for their potential utility as therapeutic or preventive vaccines. The present investigation was done with five different type of antigens produced from single pathogen. It also identifies the immune complex and DNA antigen as most suitable for vaccine development against pathogens. Antigens may vary within the host during the course of infection or antigenic types of parasites in the population. Antigenic variation is an important mechanism used by pathogenic microorganism for escaping and neutralizing activities of the antibody. Immune complexes and DNA antigen serve as vaccines, and it can be used to develop multi antigen. Furthermore, it is also easy to produce in a large scale. Vaccination is still the best way for prevention of bacterial diseases. The conditions for the preparation of antigens of intact natural composition and conformation from Escherichia coli (whole cell and heat killed), was determined using Swiss albino mice (Balb/C) as experimental species. A novel approach for vaccine design and production is discussed. Components leading to the pathology, together with the prevention of the escape mechanism involved by pathogens are the major idea of vaccine technology.

An abnormal presence of lipids in blood leads to cardiovascular disease (CVD). The  main objective of the present study was formulation of nanoparticles by using ethanolic extract of Cocculus pendulus and evaluation of hypolipidemic activity in rats fed with high fat diet. Preliminary phytochemical analysis of whole plant of Cocculus pendulus revealed the presence of various phytoconstituents like alkaloids, phytosterols, flavonoids, triterpenes and volatile oils. HPTLC fingerprint analysis of ethanolic extract of plant Cocculus pendulus reveals presence of 13 peaks and 10 peaks in 256 nm and 366 nm respectively and GC-MS shows presence \(\beta\)-Sitosterol as a marker compound. Nanotechnology is a promising technique to increase the bioavailability of herbal medicines. The formulation of nanoparticles of ethanolic extract of Cocculus pendulus by using Solvent evaporation technique was employed for the preparation of nanoparticles with different polymers concentration and finally the optimized nanoformulation demonstrated a mean particle size, polydispersity index, zeta potential are 536 nm, 0.256, and - 16.4 mV respectively. Antiatherogenic activity was performed on male Wistar rats with 7 groups 6 animals each with doses of ethanolic extract 200, 400 mg/kg and optimized nanoparticles with 40, 80 mg/kg Effect of ethanolic extract of Cocculus pendulus and  nano formulations with two different doses on 8 weeks with High fat diet were investigate. Finally results shows that per oral route of administration of Cocculus pendulus and Nano formulations at two doses level showed significant reduction in inclined levels of body weight, total cholesterol, low density lipoprotein, phospholipids, triglycerides, VLDL levels and similarly significant increase in high density lipoproteins levels. Hypolipidemic activity of  Cocculus pendulus nanoparticles with dose of 40 mg/kg and 80mg/kg are more effective than the ethanolic extract Cocculus pendulus at a dose of 200 mg/kg, 400 mg/kg.

Objective: The goal of this study was to prepare a bilayer tablet of atenolol for biphasic drug release in order to improve its bioavailability and absorption in the lower gastrointestinal tract.

Methods: Croscarmellose sodium and sodium starch glycolate were used as super disintegrants in the formulation of immediate release crosspovidone, and it was directly compressed. Different grades of HPMC k4 M, HPMC K15 M, Gum Tragacanth, Gum Acacia, Guar gum, and Ethyl cellulose are used for the sustained release portion. Prior to compression, preformulation studies were carried out. Using a USP dissolution apparatus type 2(paddle), the compressed bilayer tablets were evaluated for weight variation, dimension, hardness, friability, drug content, disintegration time, and in-vitro drug release .

Results and Conclusion: The formulation IR3 showed regression coefficients value (r²) is 0.994. The data suggested that release kinetics follow first order drug release. IR3 formulation showed 95% drug release in 30 min and regression coefficients value (r2) value was found to be 0.994. It has 5%w/w crosspovidone used. The formulation F9 showed regression coefficients value (r²) is 0.992. The data suggested that release kinetics follow Zero order drug release. F9 formulation, using HPMC K15M and Gum acacia (1:1) showed 91.20% drug release at the end of 12 hrs. Formulation IR3F9 showed drug release for bilayer tablet faster release which containing 5%w/w crospovidone in immediate release layer and Sustained release which containing HPMC and guar gum (1:1). Formulation IR3F9 showed swelling index was found to be 206%, floating lag time was found to be 2 min. and total floating time up to 12 hrs. Dissolution profile for Bilayer tablet IR3F9 formulation a good relation between cumulative drug release and time. The release pattern showed 95.23% up to 12 hrs.

A Nanosponge is a modern and evolving technology that provides both topical and oral drug delivery in a targeted and controlled way. Nanosponges are made from nano-polymer spheres that can suspend or trap several substances in a product that is formulated, such as gel, cream, ointment, fluids, or powder. They are then applied to a formulated product. This technology provides ingredient interlocking and decreases side effects, enhances stability, beauty, and improves versatility in formulations. The advanced distribution of medicines is Nanosponge. It is a basic support device for the targeted delivery, lipophilic or hydrophilic, of all types of drugs. The phrase 'nanosponge' was originally used in the nineties to overcome the limits of the indigenous cyclodextrins (CD), and especially their water solubility by its nanopore, spongy-like structure. In the years after CD NS has been introduced, attempts were made to explain how low molecular weight molecules, high molecular weight, hydrophobic or hydrophilic nature can host their behavior and function and understand their ability to tolerate cyclodextrin changes, cross-connection, and cross-connection molecules. They have made great strides in different fields, such as agribusiness, pharmaceuticals; biomedicine, and biotechnology, and there is no full NS analysis. This analysis offers a summary of the science of the CD NS, which focuses on historical roots and main issues from fairly simple networking's in the 1960s to multifunctional polymers in the last 50 years. The approach adopted in this study was to examine the historic trends in the NSs so that through the analysis of various authors and their reviews, Today, they might understand their role and imagine their future.

Quantitative structure activity relationship (QSAR) study was performed on a series of 3,5-diaryl-4,5-dihydro-1H-pyrazole-1-carbothioamides in order to establish quantitative relationship between epidermal growth factor receptor (EGFR) kinase inhibitory activity and physicochemical / structural properties. Several statistical regression equations were obtained using multiple linear regression (MLR) analysis. Most statistical significant model generated (Model 2), justifies 72% (r2 = 0.7194) of the overall variance in the training set as well as it has internal (q2) and external (pred_r2) predicative ability of ~ 65% (q2 = 0.6472) and 75% (pred_r2 = 0.7530) respectively. This model is further validated and Mean Absolute Error (MAE)-based criteria applied on 95% data showed that MAE+3* Standard Deviation (SD)[= 0.3359 < (0.25 * TrainingSetRange = 0.55250)], indicate that projection quality of the model is good. In this model positive coefficient value of T_C_C_4 [This is the count of number of carbon atoms separated from any carbon atom (single, double or triple bonded) by 4 bond distance in a molecule] and Most+ve&-ve Potentail Distance [This descriptor signifies the distance between points having the highest value of +ve and highest value of –ve electrostatic potential on van der waals surface area of the molecule] on the biological activity indicated that higher value leads to enhanced epidermal growth factor receptor (EGFR) kinase inhibitory activity whereas lower value leads to decrease activity. Contribution chart reveals that the descriptors T_C_C_4 and Most+ve&-ve Potentail Distance contributing 75.97% and 24.03% respectively.

Tape method is most commonly used and is considered as the standard and most commonly used method for measuring spine mobility. The goniometric method, although not the most accurate, seems to be clinically accessible, objective and easy to use of measuring spinal mobility but not many studies have used and tested it for measuring spine ROM in healthy subjects. The chapter consists of discussion on specific, sensitivity and accuracy of goniometer compared with tape method for measuring spinal mobility in all planes (sagittal, frontal and transverse) in healthy adults. it also recommends method which can be used for measuring each movement of spine with their possible reasons.

Objective: The study's intention was to develop floating in situ gel formulations of Ciprofloxacin that have a narrow absorption window and are primarily absorbed in the proximal areas of the GIT. These formulations prolong the targeted action on bacteria and can be used to treat Helicobacter pylori (H. pylori) infections and urinary tract infections.

Methods: In situ gel formulations were created by varying the concentrations of sodium alginate, a bio-degradable polymer that forms in situ gels, and calcium carbonate, a cross-linking agent. Physical appearance, pH, in vitro drug release, viscosity, in vitro floating behavior, in vitro gelling capacity, and drug content were all evaluated in the formulations. Ciprofloxacin, excipients, and formulation optimization were all subjected to FTIR analysis.

Results: All of the formulations demonstrated optimal viscosity, allowing for easy administration and swallowing. All formulations floated for more than 12 hours with a floating lag time of 32-70 seconds. The in vitro gelling capacity increased as the polymer and gelling agent concentrations were increased. The rate and extent of drug release were reduced as polymer concentration increased. Among all formulations, F4 containing 4%w/v sodium alginate and 4%w/v calcium carbonate demonstrated sustained in vitro drug release (95.6%) over a 12-hour period. FTIR studies revealed that there was no interaction between the drug and the excipients used. The formulations' drug release followed first order kinetics with Fickian diffusion.

Conclusion: Ciprofloxacin was successfully developed as a pH-triggered floating in situ gelling system using sodium alginate.  

Plants are major source of human living. Since the beginning of the era plants have been used for medicinal purposes. There is an urgent need to investigate the mechanisms of chemical constituents in plants, particularly saponins and cardiac glycosides, because of their ability to save damaged cells in cardiac muscle. Articles describing the cardioprotective mechanism of medicinal plants were found by searching databases such as Google Scholar, Medline, PubMed, and the Directory of Open Access Journals. Saponin, flavonoids, glycoside, steroid, alkaloids, tannin, phenol, phlobatanin, terpenoids, and anthraquinone are chemical constituents in plants that improve cardioprotective activity and reduce cardiac abnormalities. The current review article provides information on the use of medicinal plants, specifically against cardiac diseases, as well as a study of molecules/phytoconstituents as plant secondary metabolites for their cardioprotective potential. The main objective of this review chemical constituents in herbal or traditional plants can be lifesaving in chronic heart diseases.

Aim: Series of bioactive sulphonamides were synthesized using threonine an essential amino acid and their antioxidant and antimicrobial activities were evaluated.

Methodology: The reaction of L-threonine with sulphonyl chloride afforded sulfamoyl carboxylic acids while their carboxamide derivatives were obtained via ammonolysis. The structures of the synthesized compounds were elucidated using FTIR, ¹H-NMR, ¹³C-NMR and elemental analysis. The antioxidant and antimicrobial activities were determined In vitro and in silico.

Results: it was observed that the best in vitro antibacterial activities against Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Salmonella typhi, Pseudomonas aeruginosa, and antifungal activities against Aspergillus niger and Candida albicans were exhibited by compounds 1b and 1d. Compound 1f (IC₅₀ = 1.150±0.003µg/ml) displayed the best antioxidant activity. Compound 1a showed a higher antibacterial binding energy (-11.51 kcal/mol) than penicillin (-10.89 kcal/mol) while 1c had an antifungal binding energy (-10.48kcal/mol) comparable to ketoconazole (-10.85 kcal/mol).

Conclusion: The sulfonamide derivatives synthesized from threonine were found to be potential antimicrobial and antioxidant agents.

The first objective of this project is to synthesize innovative cyclic peptides and test them as Epidermal Growth Factor Tyrosine Kinase Inhibitor (EGFR-TKI) by Enzyme-Linked Immuno Sorbent Assay (ELISA) experiment. The Cyclic Phe-Phe-Phe-Gly Tetrapeptide (CPTP) exhibited a strong IC₅₀ of 55.6 nM while cyclic heptapeptides, \(\mu M\) level. The second objective is to compare by AutoDock experiment the effectiveness between our most potent CPTP and US Food Drug Administration (FDA) approved erlotinib. The result of a docking study of the CPTP showed a fairy strong inhibition free energy of -7.74 kcal/mol. The two hydrogen bonds were observed between the donor hydrogen of CPTP and the acceptor oxygen of the EGFR residue PHE771, confirming the strong affinity between CPTP and EGFR protein. Cancer reveals the major mortality world-wide. The comprehensive understanding and strategy of cancer is inevitable for the diagnosis, treatment, and prevention of cancer. Various therapeutic techniques aimed at overcoming the resistance to currently available EGFR inhibitors and preventing its onset failed. Our honest goal is that our novel tetracyclic peptide will be a safe and effective treatment for non-small-cell lung cancer without causing resistance. We hope that the future clinical research will elucidate the fact that our novel CPTP is a resistant free and effective drug for the treatment of Non-Small-Cell (NCS) lung cancer. 

Aim: Although the polyherbal siddha formulation veppampoo Mathirai is effective at lowering blood pressure, its safety is unknown. Heavy metals, aflatoxins, pesticide residue, and microbial count have not yet been evaluated. The parameters listed above were evaluated in the current study. The present study was aimed to evaluate the safety parameters (heavy metal, aflatoxin, pesticide residue and microbial profile) of Veppampoo Mathirai and the objective was to determine whether the above parameters of VPM were within normal limits as per AYUSH guidelines.

Materials and Methods: According to AYUSH [Ayurveda, yoga, unani, siddha, naturopathy] Pharmacopoeial laboratory for Indian medicine (PLIM) guidelines, the formulation was evaluated for its safety parameters at Noble research solutions, kolathur, Chennai, accredited with ISO 9001: 2015. Heavy metals were tested using an Atomic Absorption Spectrometer (AAS), and aflatoxins were tested using Thin Layer Chromatography (TLC). Pesticide residues were determined using GC/MS, and microbial counts were determined using the pour plate method.

Results: The presence of heavy metals mercury, arsenic, lead, and cadmium was found to be within the recommended limit as per AYUSH Pharmacopoeial Laboratory for Indian Medicine Guidelines, while Aflatoxin, pesticide residues, and microbes were not found in the sample, indicating that the formulation Veppampoo Mathirai (VPM) was toxicity-free.

Conclusion: According to AYUSH's PLIM criteria, heavy metal level was below the acceptable limit in VPM. Aflatoxins and pesticide residues were not found in the current batch of VPM, whereas microorganisms and particular pathogens were not present. As a result, the current research confirms that the formulation is safe for therapeutic use.

This paper includes the most relevant analytical methodology used for the determination of Cefadroxil antibiotic.

Cefadroxil is an orally active semi-synthetic \(\text { B-lactam }\)  antibiotic from the cephalosporin group, characterized by its prolonged action. This compound is effective against susceptible bacteria causing infections of the urinary tract, skin and soft tissue as well as pharyngitis and laryngitis. A variety of analytical methods have been proposed for the determination of cefadroxil in biological fluids and pharmaceutical samples. The spectrophotometric method using oxidative coupling reaction has been performed in which chloranillic acid & MBTH has been used as reagent at 535 nm & 420 nm absorbance with linear range of 15-415  \(\\mu \mathrm{g} / \mathrm{ml} \& 1-12 \mu \mathrm{g} / \mathrm{ml} .\)  Speed of analysis has become of paramount importance in many application areas, such as in pharmaceutical and clinical analysis, in order to increase throughput and reduce costs. In recent years, it is becoming a powerful analytical tool for pharmaceuticals in general and also for CFL determination because of the low detection limit and wide linear dynamic range. In this way, the recently invented methods offer new practical possibilities for increasing their efficiencies.

Chemistry or Medicinal chemistry is a discipline at the Interchange ofchemistry and pharmacology engaged with designing, methodize andcreating drug chalcones is a conventional term given tointensifies bearing the (E)- 1,3-diphenylprop-2-en-1-one, which can befunctionlized in the propane chain by the presence of olefinic, keto as well ashydroxyl group.Chalcones are a significant class of regular items and are considered as the forerunners of flavonoids and isoflavonoids. Synthetically, is (E)-1,3-diphenylprop-2-en-1-one in which two aromatic ring rings are joined by a three carbon bond having a carbonyl moiety and \(\alpha, \beta\) unsaturation. The compounds with the backbone of chalcones have been accounted for to have different pharmacological activities for example, antimicrobial, anti- inflammatory, analgesic, anti-platelet, anti-ulcerative, antimalarial, anticancer, antiviral, anti-leishmanial, antioxidant, antitubercular, anti-hyperglycemic, immunomodulatory, inhibition of chemical mediators release, inhibition of leukotriene B4, inhibition of tyrosinase and inhibition of aldose reductase activities. This paper mainly focuses on chalcones synthesized by Claisen Schmidt condensation which involves the condensation between an aromatic aldehyde or ketone with an aliphatic ketone or aldehyde catalysed by the presence of dilute alkali or acid to form alpha beta unsaturated compound. through reviewing different biological significance of chalcones andtheir derivatives have been reported along with their antibacterial movement against, Bacillus pumilis, Bacillus subtilis (gram-positive) and Escherichia coli, Proteus vulgaris (gram-negative). The antiinflammatory action of the sixteen chalcones has been assessed byutilizing carrageenan-actuated rodent paw oedema strategy . 

Aim: The objectives of this work is to develop and validate a simple, specific, accurate, and precise HPLC method for simultaneously estimating Curcumin (CUR) and Thymoquinone (THQ) in bulk and formulation, in accordance with ICH principles for analytical method development and validation.

Study Design: Developing an RP-HPLC method for estimating Thymoquinone and Curcumin in bulk and formulation utilizing a C-18 Inertsil column and optimizing variables.

Place and Duration of Study: The current study was conducted at Shri D. D. Vispute College of Pharmacy and Research Center, Panvel in year 2021.

Methodology: The RP-HPLC method was developed using an isocratic mobile phase consisting of acetonitrile and water in a ratio of (82:18) v/v, at a flow rate of 0.9 mL/minute over Inertsil ODS, 250× 4.6 mm, 5 µm column, at 30°C column oven temperature. Detection was done using a photodiode array at 256 nm.  

Results: Curcumin and thymoquinone had retention times of 3.5 and 4.3 minutes, respectively. For thymoquinone and curcumin, the method demonstrated excellent linear response in concentration ranges of 4-18 \(\mu \mathrm{g} / \mathrm{ml}\) and 10-45 \(\mu \mathrm{g} / \mathrm{ml}\), respectively, with correlation coefficient (R2) values of 0.999 for both. System precision and method precision studies were less than the maximum allowable limit percentage of relative standard deviation \(\leq 2.0 \text { i.e., } 1.61 \%\) and 1.62 % for curcumin and 0.47 % and 0.42 % for thymoquinone respectively. Both medications' mean percent recovery was within acceptable ranges. Because all of the results satisfied the acceptance criteria, the designed and validated HPLC method is simple, accurate, precise, and acceptable for analysis.

Conclusion: The developed RP-HPLC method at single wavelength was verified according to ICH guidelines for system appropriateness, specificity, linearity, accuracy, precision, and robustness, and can be used for routine quality monitoring of drugs in pharmaceutical dosage forms. This method can be employed for routine quality control analysis of curcumin and thymoquinone samples simultaneously.  

Background: Hepatic disorders or degeneration remain an important health problem till today as liver is the major organ of biotransformation for xenobiotics. Even so many antioxidant and hepatoprotectives are available, once the degeneration of liver occurred beyond the repair capability results in liver failure and death.

Objective: To study the hepatoprotective activity of an aqueous and alcoholic extract of Acorus calamus rhizomes against the paracetamol induced hepatotoxicity in rats.

Methods: Hepatotoxicity was induced by oral administration of paracetamol and chemical parameters such as Glutathione peroxidase, Glutathione reductase, Glutathione, Catalase, lipid peroxidation and histopathological changes in liver was studied by comparing with Silymarin, a standard hepatoprotective drug.

Results: Treatment of rats with aqueous and alcoholic extract of Acoruscalamus rhizome after paracetamol administration normalized the altered levels of above parameters which may comparable with Silymarin and Vit-E. The hepatoprotective activity was confirmed by histopathological examination of the liver tissue of control and treated animals.

Conclusion: Based on the result it can be concluded that Acorus calamus rhizome possesses hepatoprotective effect against paracetamol-induced hepatotoxicity in rats. The use of Acoruscalamusmay be recommended for lifestyle-related diseases including hepatic diseases and also to improve the general health condition in animals including human beings. Extensive studies in target species should be preceeded before the clinical application of Acoruscalamus as an antioxidant agent.