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Item Investigation of Anticancer and Antibacterial Metabolites from Seaweeds of the Bay of Bengal and their Associated Endophytic Fungi(© University of Dhaka, 2026-03-02) NOOR, SADIAThe relentless pursuit of novel anticancer and antibacterial agents has directed significant research toward marine ecosystems, with seaweeds and their endophytic fungi emerging as exceptionally promising reservoirs of bioactive metabolites. This research aimed to systematically investigate the bioactive potential of marine endophytic fungi derived from seaweeds of the Bay of Bengal, Bangladesh. Three seaweed species—Ulva sp. (Chlorophyta), Gracilaria sp. (Rhodophyta), and Sargassum sp. (Phaeophyta)—were collected, morphologically identified, and served as hosts for the isolation of endophytic fungi on potato dextrose agar (PDA) media. Twelve fungal strains were isolated and identified through a polyphasic approach combining macroscopic/microscopic morphological characterization and molecular phylogenetic analysis of ITS sequences. The fungal strains comprised Chaetomium globosum (UE-1, SE-1), Nigrospora magnoliae (UE-2), Curvularia sp. (UE- 3), Curvularia moringae (UE-4), Aspergillus terreus (UE-5, GE-2, SE-2), Collariella gracilis or C. virescens (UE-6), Aspergillus subversicolor (GE-1), Cladosporium halotolerans (GE-3), and Curvularia perotidis (SE-3). Critically, all twelve endophytic fungal strains were reported for the first time from the Bay of Bengal, Bangladesh, with Nigrospora magnoliae being reported as a fungal endophyte for only the second time worldwide. Preliminary chemical profiling of crude extracts via Thin Layer Chromatography (TLC) and Gas Chromatography-Mass Spectrometry (GC-MS) indicated a rich diversity of secondary metabolites, including steroids, terpenoids, flavonoids, and fatty acid derivatives. Bioassay-guided fractionation led to the isolation of twelve pure compounds using various chromatographic techniques, and their structures were elucidated through 1D and 2D NMR spectroscopy. From C. globosum, six compounds were isolated: the new natural compounds UC-71 [(E)-1-(2′,4′- dihydroxy-3′,5′-dimethylphenyl)-2-buten-1-one] and UC-58 [1-(3′,4′-dihydroxy-2′,5′- dimethylphenyl)-3-hydroxybutan-1-one], alongside known metabolites chaetoviridin E (UC-59), a mixture of chaetoviridin A epimers (UC-60), chaetoglobosin G (UC-62), and chaetoglobosin B (UC-63). Another six compounds were isolated and characterized for the first time from N. magnoliae: ergosta-4,6,8(14),22-tetraene-3-one (UN-172), sterigmatocystin (UN-171), dihydrosterigmatocystin (UN-176), 11-oxo-(9E)- octadecenoic acid (UN-233), ostopanic acid (UN-235), and cerevesterol (UN-411). Bioactivity screening revealed that the crude extracts possessed significant iv antimicrobial activity against Gram-positive (Staphylococcus aureus, Bacillus megaterium) and Gram-negative bacteria (Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa), and antifungal activity against Aspergillus niger and A. flavus in disc diffusion assays, outperforming their host seaweeds which showed no activity. Antioxidant activity, evaluated via DPPH radical scavenging, was particularly potent in extracts of A. terreus strains (IC50 value as low as 7.88 μg/mL). Brine shrimp lethality bioassay indicated strong cytotoxicity (LC50 value less than 30 μg/mL) for numerous extracts. The isolated pure compounds also exhibited notable bioactivities; for instance, UC-60 showed remarkable antifungal activity against Candida albicans (31.3 mm zone at 100 μg/disc), and several compounds demonstrated significant cytotoxicity in subsequent MTT assay on the HeLa cervical cancer cell line. The MTT assay revealed that compound UN-235 exhibited significant dose-dependent cytotoxicity against HeLa cells (IC50 value 243.04 μg/mL), demonstrating potency comparable to doxorubicin, thereby identifying it as a promising cytotoxic candidate worthy of further investigation. These comprehensive findings underscore the immense, largely untapped potential of endophytic fungi from Bangladeshi seaweeds as prolific sources of novel chemotherapeutic agents, with the isolation of new natural compounds and first-reported metabolites highlighting their critical value in anticancer and antibacterial drug discovery pipelines.Item Chemical and pharmacological profiling of Jatropha pandurifolia and Syzygium reticulatum(© University of Dhaka, 2025-02-19) JAHAN, NISRATJatropha pandurifolia Andrews (family: Euphorbiaceae) and Syzygium reticulatum (Wight) Walp. (family: Myrtaceae) were studied for their secondary metabolites and their biological activities were evaluated both in vitro and in vivo. Through extensive NMR experiments—including ¹H NMR, ¹³C NMR, and ¹H-¹H COSY, HSQC and mass spectrometry, a total of fifteen compounds were identified from the plant extracts. Thirteen of these chemical compounds were isolated specifically from the stem bark of J. pandurifolia including 2-epi-jatrogrossidione, integerrimene, jatrophatrione, citlalitrione, dotriacontyl trans-ferulate, triacontyl trans-ferulate, octacosyl trans-ferulate, hexacosyl trans-ferulate, octacosyl cis-ferulate, β-sitosterol, stigmasterol, n-dotriacontanol, and 1,2-dioleoyl-3-palmitoyl-glycerol. Dotriacontyl trans-ferulate was isolated and characterized as a previously unreported molecule. Bis(2-ethylhexyl) phthalate and stigmast-4-en-3-one were isolated from the leaves of S. reticulatum, marking the first report of these compounds from this plant species. In vivo investigation was carried out to assess CNS depressant, antidiarrheal, and analgesic activities, while in vitro study was assessed for antioxidant, cytotoxic, and thrombolytic properties. Furthermore, in silico investigation of selected pure compounds were conducted using molecular docking and ADMET (absorption, distribution, metabolism, excretion, and toxicity) profiling. CNS depressant activity was carried out by open field and hole cross methods at 100 mg/kg, 200 mg/kg, and 300 mg/kg, where diazepam was used as standard. In open field and hole cross method, highest inhibition of movements were 87.37% (***P<0.001) and 84.78% (*P<0.05), respectively for methanolic extract of J. pandurifolia leaf at 300 mg/kg. Furthermore, at the same dose highest inhibition of locomotor activity were 73.68% (***P<0.001) and 80.43% (**P<0.01) for ethyl acetate extract of S. reticulatum leaf in aforementioned methods. Diazepam exhibited 91.58% and 95.65% movement inhibition, respectively. The antidiarrheal effect was performed using the castor oil induced diarrheal model at 200, 400, and 600 mg/kg, using loperamide as the standard. The frequency of diarrhea was reduced by 85.95% at 600 mg/kg of Jp-ML, where loperamide showed 98.57% gastric inhibition. At the same dose, Sr-EAL exhibited a delay in diarrheal onset of 87.53% where loperamide showed 94.84% gastric inhibition. The analgesic activity was assessed by acetic acid induced writhing ix method and formalin induced nociception test. Jp-ML showed the highest dosedependent inhibition of writhing by 66.67% (**P<0.01) at 300 mg/kg, whereas indomethacin showed 78.43% pain reduction. In the formalin test, pain inhibition were 63.92% (***P<0.001) in the early phase and 70.35% (***P<0.001) in the late phase at the same dose. On the other hand, Sr-EAL showed the highest inhibition of writhing by 70.59% (***P<0.001) at 300 mg/kg. Inhibition of paw licking were recorded by 54.55% (***P<0.001) and 70.52% (***P<0.001) in the early phase and late phase, respectively. Antioxidant activity was estimated using the DPPH free radical scavenging assay, with BHT as the standard. The lowest IC50 value were 8.11 μg/mL and 10.34 μg/mL for JP-ML and Sr-EAL, respectively. Pure compounds (JP- 10, JP-13, JP-25, and JP-460) exhibited promising antioxidant activity, with highest antioxidant activity of JP-25 (IC50 =13.10 μg/mL). Cytotoxicity was assessed using the brine shrimp lethality assay, with vincristine sulphate as the standard. The LC50 values were 1.60 μg/mL for Jp-EAL and 1.70 μg/mL for Sr-ML. Among the four pure compounds tested, JP-460 exhibited the strongest cytotoxicity, with LC50 value of 1.27 μg/mL. Cytotoxicity test was also carried out on the HeLa cell line both qualitatively and quantitatively. In qualitative test JP-13 and JP-460 showed a cell survival rate of over 95%, whereas JP-200 resulted in a cell survival rate of only 5%, indicating strong cytotoxicity. In quantitative MTT assay, JP-200 exhibited LD50 value of 272.45 μg/mL. Thrombolytic activity was evaluated using streptokinase as standard. The highest clot lysis activity were 63.32% and 67.58% for the Jp-ML and Sr-EAL, respectively. JP-460 demonstrated significant clot lysis activity, with 71.04% clot rupture. Finally, molecular docking studies revealed that compound JP-460 exhibited strong binding affinity (-9.4 kcal/mol) and binding efficacy (0.41 (kcal/mol per non-hydrogen atom) to the human GABA-A receptor. The ADMET assessment confirmed that the isolated compounds have preferable pharmacokinetic and safety profiles, supporting their potential for therapeutic application.Item The potentiation and broadening the effect of low molecular hypoglycemic drugs through interaction with some CNS stimulant molecules(© University of Dhaka, 2025-03-10) Mohiuddin, MohammadItem Cytotoxic and antimicrobial constituents from some Bangladeshi Bridelia and Erythrina species(© University of Dhaka, 2025-03-10) Anjum, AdeebaItem Development of sustainable method for the Quality Control of traditional medicines on the basis of phytoequivalence and chemical fingerprinting(©University of Dhaka, 2023-12-07) Koly, Sabiha FerdowsyBackground: Herbal drugs are composed of single or several types of medicinal plants with additives. This type of preparation is gaining a wide range of popularity among a large a number of people throughout the world. With the increase of usage, it is necessary to maintain the quality of these drug preparations. Therefore, analytical approaches for their intended use in drug quality evaluation need to be validated. This study was planned to develop simple, rapid, selective, precise and economical method for the quality control of herbal preparations. Methods and Materials: Infrared (IR) spectroscopy was used to determine phytoequivalence among the samples. This work outlined a method for identifying herbal drugs on the basis of phytoequivalence. However, as this is a time-consuming and expensive process, the current analysis did not attempt to identify the specific chemical compounds found in the herbs. The marked bands served as a comparative tool for herbal plants and the medications made from them. The quantitative method of comparison was applied for the comparison of crude herbal standard with herbal sample containing formulations. Thin Layer Chromatography (TLC) was used as another analytical method for identification and showing phytoequivalence. It has mostly been used for the qualitative analysis of herbal medicines and to characterize and track the components visually or as an initial separation technique. Single or multiple herbs containing preparation can be analyzed using ultraviolet (UV) spectroscopy. In this technique, it involves the measurement of ultra violet radiation absorption by the substance dissolved in solution. Both qualitative and quantitative analysis can be done through this technique. In the present study, it was aimed at development and validation of UV-spectroscopic technique according to International Conference on Harmonization guidelines which is known as ICH (Q2) guidelines for the analysis of herbs containing polyherbal formulation. The method validation parameters like specificity, precision, accuracy, linearity, range, repeatability and robustness were studied according to ICH (Q2) guidelines. Results and Discussion: Amlaki (Phyllanthus emblica) showed the presence of prominent peak and maximum absorption at 303 nm. The detector response for the Amlaki was linear over the selected concentration range of 1 to 5 μg/mL with a correlation coefficient of 0.998. The absorbance values for intraday precision, found for 1 μg/mL, 3 μg/mL and 5 μg/mL were 0.0082, 0.234 and 0.396 having %RSD of 0.998%, 0.0080% and 0.0058%, respectively. The absorbance values for intermediate precision, found for 1 μg/mL, 3 μg/mL and 5 μg/mL were 0.0084, 0.234 and 0.398 having %RSD 0.6846%, 0.0137% and 0.0038%, respectively. The absorbance value for repeatability was 0.084 having %RSD 0.6901. The accuracy was between 99.348% and 101.478%. Robustness of the method was studied. The %RSD for analyst to analyst variation was 0.4851% and instrument to instrument variation was 0.9726%. The assay results of Amlaki were about 86.588%, 82.150% and 90.828% for three market preparations A, B and C, respectively, indicating insignificant interference from the other ingredients in the formulation. Black plum (Syzygium cumini) showed the presence of prominent peak and maximum absorption at 279 nm. The detector response for the S. cumini was linear over the selected concentration range of 0.1-0.5 μg/mL with a correlation coefficient of 0.9914. The absorbance values for intraday precision found for 0.1 μg/mL, 0.3 μg/mL and 0.5 μg/mL were 0.234, 0.432 and 0.735 having %RSD 0.9245%, 0.5751% and 0.4668%, respectively. The absorbance values for intermediate precision found for 0.1 μg/mL, 0.3 μg/mL and 0.5 μg/mL were 0.233, 0.432 and 0.736 having %RSD 0.8921%, 0.8346% and 0.3421%, respectively. The absorbance value for repeatability was 0.225 having %RSD 0.7950%. The accuracy was between 99.647% and 101.943%. Robustness of the method was studied. The %RSD for analyst to analyst variation was 0.8251% and instrument to instrument variation was 0.3609%. The assay results of S. cumini were about 83.152%, 86.821%, 90.082% and 80.579% for four market preparations A, B, C and D, respectively, indicating insignificant interference from the other ingredients in the formulation. During the method development phase, a number of solvents were used. Among them, methanol was selected in analysis of Amlaki and ethanol was selected in analysis of black plum. These solvents were selected as these solvents satisfied all the conditions relative to peak quality and non-interference at the specified wavelength. The wavelength of maximum absorption (λ max ) was found to be 303 nm and 279 nm in Amlaki and Black plum, respectively. As the values of %RSD is <2%, the method is validated according to ICH (Q2) guidelines. Conclusion: It can be capitulated that this method can be conveniently employed for routine quality control analysis of herbal drugs in bulk drug and other formulations.Item Design, development and evaluation of formulations of poorly water soluble drugs intended for oral delivery(©University of Dhaka, 2023-11-22) Rahman, Md. RezowanurLedipasvir and Daclatasvir, belonging to the BCS class 2, and Velpatasvir, belonging to the BCS class 4, are directly acting anti-viral agents used to treat Hepatitis C virus infections. Owing to poor aqueous solubility and oral bioavailability, development of effective delivery system for these drugs has been enormously challenging. Moreover, suitable dosage forms for pediatric and geriatric patients and patients having difficulty in swallowing as well pose added burden. Therefore, the aim of the present study was to develop a nanosuspension, via solid dispersion technique, based liquid oral suspension using Quality by Design (QbD) approach. Primarily, the compatible polymers for Ledipasvir were screened using FTIR and DSC, and finally the polymers - Poloxamer 188, Poloxamer 407, HPC and HMPC were selected, considering their ability to convert the API into amorphous state in solid dispersions. Design of formulation and analysis with the DOptimal design using Design Expert ® software revealed that Poloxamer 188 and Poloxamer 407 in 0.3:0.7 ratio of Ledipasvir:Polymer produced the optimized nanosuspension formulations with a statistically significant mathematical model. Subsequently, the formulations were stabilized using suspension vehicle optimized via Box Behnken Design using the amount of xanthan gum (gm), avicel ® RC-591 (gm) and citric acid monohydrate (gm) as independent variables whereas viscosity (cp) and zeta potential (mv) as responses. The dissolution profiles revealed that the prepared suspensions of Ledipasvir had much faster dissolution than pure API, suspensions prepared with micronized and nonmicronized API, and the market products available as tablet dosage form. In-vivo simulation studies using PKSolver ® suggested that the absorption of drug from the formulated suspensions was comparable to that of market product up to single dose level (90mg) and superseded in triplicate dose level (270 mg). The formulated suspensions were found to be stable over three- and six-months periods, identified via accelerated stability studies. Interestingly, dissolution profile of the stabilized suspensions was found to be similar after six months. An RP-HPLC method to determine the assay content of Ledipasvir in the finished product has also been developed using 3 2 full factorial design with a Diphenyl column (250 mm X 4.6 mm, 5 µm), the detection wavelength of 330 nm and the injection volume of 20 µL. The optimized method consisted a mobile phase of buffer:acetonitrile at 48:52 ratio and flow rate of 1.7 ml/min. A simple and rapid UV method was developed simultaneously to analyze Ledipasvir and shown to be equivalent to the developed RP-HPLC method. To determine the content of the residual solvent in Ledipasvir solid dispersions, a GC method was developed using the same 3 full factorial design and fused silica GC capillary column (30-m x 0.32-mm x 1.8-µm), Nitrogen with 14.0 psi through head space as carrier gas. Validation of all the developed methods were carried out by following ICH Q2 (R1) guideline. The best results were found with Poloxamer 188 in the Ledipasvir study at a drug:polymer ratio of 0.7:1.3 in terms of in-vivo simulation. Therefore, both Daclatasvir and Velpatasvir were further studied to develop solid dispersion based nanosuspensions and finally a stabilized oral suspension using Poloxamer 188. In case of Daclatasvir, the drug failed to produce amorphous solid dispersion and hence, was not further evaluated for nanosuspension preparation. On contrary, Velpatasvir produced amorphous solid dispersion and thus, nanosuspension was prepared using the same approach applied for Ledipasvir. Afterward, the nanosuspension of Velpatasvir were stabilized using the same method used to stabilize Ledipasvir nanosuspension. The study of the dissolution profiles revealed that stabilized suspension of Velpatasvir had much faster dissolution than its market product available as tablet dosage form. Finally, in-vivo simulation study revealed that single dose of formulated suspension gave the comparable absorption profile to that of the market product.Item Phytochemical and pharmacological evaluation of Pongamia pinnata, Sesbania grandiflora and associated endophytic fungi(©University of Dhaka, 2023-06-11) AFRIN, SEAGUFTASesbania grandiflora (L.) Pers and Pongamia pinnata (L.) Pierre, belonging to the Fabaceae family, have been investigated for isolating new natural compounds with promising pharmacological activities. A total of tweenty five compounds have been isolated from these two plants and endophytic fungi associated with Sesbania grandiflora. The isolated compounds were identified by extensive analysis of high resolution NMR ( 1 H-NMR, 13 C-NMR, 1 H- 1 H COSY, HSQC and HMBC) experiments and the molecular weights were determined by ESI mass spectrometry. The stem bark of Sesbania grandiflora afforded fourteen natural compounds namely, Sesbagrandiflorain A, (-) maackiain, medicarpin, sativan, 4-hydroxy-2methoxybenzaldehyde, nobiletin, kaurenoic acid, betulinic acid, lupeol, β-amyrin, stigmast-4-en-3-one, stigmasta-4, 22-dien-3-one, stigmasterol and linoleic acid. Among them, eight compounds like (-) Maackiain, 4-hydroxy-2methoxybenzaldehyde, nobiletin, kaurenoic acid, β-amyrin, stigmast-4-en-3-one, stigmasta-4, 22-dien-3-one and linoleic acid were isolated for the first time from Sesbania grandiflora. Five flavonoids and lupeol were isolated from Pongamia pinnata and were identified as pongachromene, kanugin, demethoxykanugin, 5’methoxypongapine and karanjin. Seven different endophytic fungi were isolated and identified from the plant Sesbania grandiflora, such as Fusarium solani, Fusarium fujikuroi, Fusarium equisetti, Xylaria bambusicola, Cladosporium sp, Clonostachys rosea and Colletotrichum aeschynomenes. Two endophytic fungal extract were also investigated for chemical constituents and anhyrofusarubin, bostrycoidin and myristic acid were isolated from Fusarium solani and 3β, 5α, 9α-trihydroxy-ergosta-7,22diene-6-one, 3β,5α-dihydroxy-6β-acetoxy-ergosta-7,22-diene and 3β, 5α,dihydroxyergosta-7, 22-diene-6-one were isolated from Xylaria bambusicola. All these compounds were reported for the first time from the associated endophytic fungi of Sesbania grandiflora. Different solvent fractions of the crude extract and the pure compound Sesbagrandiflorain A, were subjected to several biological activities like antimicrobial, antioxidant, brine shrimp lethality and thrombolytic activities. Among the different fractions tested against five microorganisms, the ethylacetate soluble fraction showed the highest anti-bacterial and antifungal activity against Bacillus megaterium and Aspergillus niger with zone of inhibition of 17.67 ± 1.53 mm and 12.67 ± 0.58 mm compared to the standard, kanamycin (30.67 ± 1.15mm) and ketoconazole (28.00± 1.00 mm) respectively. The pure compound displayed mild activity against Bacillus megaterium (11.67 ± 0.58 mm). In vitro antioxidant activity was studied by DPPH radical scavenging method using butylated hydroxy anisole (BHA) as the standard. Among all the samples tested, the pure compound Sesbagrandiflorain A and crude methanolic extract showed very significant antioxidant activity with IC 50 value of 28.26 ± 0.35 μg/ml and 54.59 ± 0.09 μg/mL as compared to the standard BHA (9.21 ± 0.10 μg/ml). In brine shrimp lethality assay the ethylacetate soluble fraction and methanolic extract demonstrated significant cytotoxic activity with LC 50 of 0.68 ± 0.10 and 0.66 ± 0.04 μg/ml respectively compared to Tamoxifen (0.30 μg/ml). The dichloromethane soluble fraction exhibited promising thrombolytic activity with % of clot lysis of 54.29% as compared to the standard Streptokinase, 73.57%. Among the different extractives of P. pinnata, the methanolic extract showed highest anti-bacterial activity against Bacillus megaterium and antifungal activity against Aspergillus niger with zone of inhibition of 20.67 ±1.15 mm and 17.00 ±1.00 mm respectively compared to the standards Kanamycin (30.67 ± 1.15 mm) and Ketoconazole (28.00 ± 1.00 mm). The Highest antioxidant activity was demonstrated by pongachromene with IC 50 values of 19.69 µg/ml. (BHA, 8.70 ± 0.29 µg/ml). Very mild cytotoxic activities were found for different extracts of P. pinnata in the brine shrimp lethality assay. The isolated endophytic fungi from Sesbania grandiflora were evaluated for their antioxidant activity, cytotoxicity and antimicrobial activity. Among the seven samples tested, SGFRE-1 showed highest antibacterial activity with zone of inhibition of 22.67 mm against Bacillus megaterium as compared to the standard antibiotic kanamycin (30.67 mm) and highest antifungal activity with zone of inhibition of 19.33 mm each against A. niger and A. flavus as compared to the standard antifungal Ketoconazole (28.00 mm). Moderate antioxidant and cytotoxic activities were observed by SGFE-1 with IC 50 value of 54.06 ± 0.20 µg/ml and LC 50 value of 1.90 µg/ml as compared to the standards BHA (9.30 ± 0.06) and tamoxifen 0.34 µg/ml respectively.Item Chemical and Biological Studies of Some Litsea Species Available in Bangladesh(©University of Dhaka, 2023-02-23) Bulbul, Israt JahanIn order to discover new phytochemicals as well as searching for evidence based information of traditional therapeutic uses of medicinal plants, four species of the genus Litsea belonging to the Lauraceae family were selected in this study. The selected plants are Litsea glutinosa Lour., L. monopetala Roxb., L. deccanensis Gamble. and L. lancifolia Hook. f. Two compounds were revealed from L. glutinosa and they are 4΄-O-methyl-(2 ̋,4 ̋,-di- E-p-coumaroyl)-afzelin (LGC-26, 95) and quercetin-3-O-(2 ̋,,4 ̋,-di-E-p-coumaroyl)-α- L-rhamnopyranoside [or, 5΄-hydroxyl-(2 ̋,4 ̋-di-E-p-coumaroyl) afzelin] (LGC-45-3, 96). Both of the compounds were reported for the first time from Litsea species. Five compounds were isolated and purified from the leaf extract of L. monopetala and characterized as vomifoliol (LML 363-1, 97), α-amyrin (LML 309, 98), β-amyrin (LML 301, 99), (E)-6,7,8,9,10,11-hexahydro-8,17:10,16-di(metheno)dibenzo- [h,l][1]oxa[5] azacyclotridecine-1,4-diol (LML 339-1, 100) and (Z)-1,2,3,4,5,6- hexahydro-8,11-etheno-2,13:4,12di(metheno)benzo[h][1]-oxa[5]aza-cyclopentadecine (LML 339-2, 101) by 1H and 13C NMR, COSY, HSQC, HMBC spectral data analysis. All these five compounds are reported for the first from L. monopetala, while compounds 101 and 101 appear to be new compounds. The obtained 1H NMR spectral data and the comparison with the reference value helped us to characterize lupeol (LDC-10-3, 102) and a mixture (4:1 ratio) of β-sitosterol and stigmasterol (LDC-10-2, 55 & 56) from L. deccanensis and β-sitosterol (LLC-10-1, 55) from L.lancifolia. Lupeol has been isolated from L. deccanensis for the first time. 4ʹ -O-methyl (2 ̋,4 ̋-di-E-p-coumaroyl) afzelin (95) Quercetin3-O-(2 ̋,4 ̋-di-E-p-coumaroyl)-α-Lrhamnopyranoside [or 5΄-hydroxyl-(2 ̋,4 ̋-di-E-p-coumaroyl) afzelin] (96) The crude extracts of L. glutinosa, L. monopetala, L. deccanensis and L. lancifolia were evaluated for biological activities through in-vitro and in-vivo screenings. For antidiarrheal activity test, 100, 200 and 400 mg/kg bw methanol extract of L. glutinosa (MELG), L. monopetala (MELM), L. deccanensis (MELD) and L. lancifolia (MELL) were administered in two animal models where 0.5 ml castor oil was used for diarrhea induction and 3 mg/kg bw loperamide was used as standard drug. In all the groups treated with MELG, MELM, MELD and MELL extracts wet feces number, total number of feces and total weight of the foecal output were decreased significantly (p<0.05) with rising of doses. The maximum peristaltic inhibition was observed 32.36%, 22.52%, 26.26% and 33.22% for 400 mg/kg by MELG, MELM, MELD and β- amyrin (99) (E)-6,7,8,9,10,11-hexahydro- 8,17:10,16-di(metheno)dibenzo- [h,l][1]oxa[5]azacyclotridecine- 1,4-diol (100) (Z)-1,2,3,4,5,6-hexahydro-8,11-etheno- 2,13:4,12-di(metheno)benzo- [h][1]oxa[5]azacyclopentadecine (101) Stigmasterol (56) Lupeol (102) β-Sitosterol (55) Vomifoliol (97) α- amyrin (98) MELL extracts respectively. The peristaltic indices were 59.0%, 79.0%, 59.1% and 63.0% for 400 mg/kg of MELG, MELM, MELD and MELL respectively compared to the control (90.0%) and standard (66.7%) groups. For all the plant extracts the percentage inhibition of gastrointestinal motility and peristalsis index were comparable to the standard. In analgesic activity test, MELG, MELM, MELD and MELL on acetic acid-induced writhing in mice at two different doses (100 and 200 mg/kg bw) showed significant reduction of squirming (p<0.001, p<0.01 and p<0.05) in a dose dependent manner as paralleled to control. In the second animal model (Eddy’s hot plate method), pain was induced by heat and analgesia was assessed by counting the time required for the initiation of the reaction. Out of the four plants, all the plants at 100, 200 and 400 mg/kg doses, increased the latency time. The pain-relieving activity data (formalin method) are presented as licking and biting time in seconds at early- and late-phases of treatment with plant extracts. In both the early- and late-phase, reaction time for licking and biting hind paw were decreased with the increment of the doses (from 100 mg/kg bw to 200 mg/kg bw) but in the late phase (20-30 min) the reaction time was decreased significantly (p < 0.05) with the increment of doses for all the studied plant extracts as well as standard indomethacin at 10 mg/kg bw. The effects of MELG, MELM, MELD and MELL on blood glucose level in streptozocin (STZ) induced diabetic rats were found to drop the blood glucose level (BGL) significantly (p<0.05) after 7th days of treatment with the plant extracts at 300 and 500 mg/kg/day doses. Percentage inhibition of blood glucose level for MELG, MELM, MELD and MELL were comparable with that of standard metformin and they are 66.69%, 57.06%, 68.16% and 69.33% respectively at 500 mg/kg/day dose as compared to the untreated diabetic control group. Hole cross test was performed to investigate the possible neuropharmacological effects (CNS stimulant or depressant) of MELG, MELM, MELD and MELL and all the extracts at two different doses (300 and 500 mg/kg bw) found to reduce locomotion in the test animals and to decrease the passing number through the hole in between the hole cross chamber by the animals in a dose dependent manner. Three fractions of four different species of Litsea were studied for antimicrobial activity by disc diffusion method. The results of antimicrobial activity test of different fractions showed mild to moderate activity for L. monopetala, very good activity for L. lancifolia, moderate to good activity for L. glutinosa and mild to very good antimicrobial activity for L. deccanensis against the microorganisms selected for this study. The quantities of phenolic compounds were found in ethyl acetate fraction of L. glutinosa (103.04±0.06), followed by ethyl acetate fraction of L. lancifolia (79.94±0.07). Among the plants L. lancifolia and L. glutinosa have shown very good total phenolics compared to L. deccanensis and L. monopetala. In DPPH free radical scavenging activity test, the IC50 for pet-ether, chloroform and ethyl acetate fractions of L. deccanensis were 31.75 μg/ml, 24.62 μg/ml and 31.04 μg/ml, respectively. All the values are comparable with that of ascorbic acid (31.66 μg/ml). For L. lancifolia 63.97 μg/ml pet-ether, 65.91 μg/ml chloroform and 80.46 μg/ml ethyl acetate extractives were required for 50% scavenging of free radicals. The effective concentrations for pet-ether, chloroform and ethyl acetate fractions of L. glutinosa were measured as 25.19 μg/ml, 37.90 μg/ml and 67.41 μg/ml, respectively. The IC50 values were 31.94 μg/ml, 24.91 μg/ml and 31.10 μg/ml for pet-ether, chloroform and ethyl acetate fractions of L. monopetala, respectively. From results, it may be proposed that three different extractives of L. deccanensis, L. lancifolia, L. glutinosa and L. monopetala were able to exhibit the free radical scavenging activity compared to ascorbic acid, a potent antioxidant compound. The docking simulation was conducted against aldose reductase (AKR1B1) protein model with the purified compound 95 and 96 by using Auto Dock Vina software. Compound 95 exposes the higher negative binding affinity (-9.8 kcal/mol) as compared to the compound 96 with binding affinity (-9.4 kcal/mol) for the interaction of the target protein aldose reductase (AKR1B1). Compound 95 exhibited strong connection with eleven hydrophobic bonds, hydrogen bonds and one other bond while compound 96 developed stable interactions by three hydrogen bonds, and eleven hydrophobic bonds. During investigating interaction pattern, binding affinity, and best binding poses of the compounds it can be proposed that both structures might be promising inhibitors against aldose reductase (AKR1B1) protein. Molecular docking analysis of isolated compounds 95 and 96 (−9.4, and −8.9 kcal/mol, respectively) against human pancreatic alpha amylase showed promising docking affinity. Compound 95 formed polar contacts with Tyr-151, Thr-163, Arg-195, Asp-197, His-201, and His-299 residues and compound 96 showed polar contacts with Gln-63, Arg-195, Asp-197, and His-299 residues. These findings suggest that these compounds are promising inhibitors of human pancreatic alpha amylase. The docking results of vomifoliol clearly indicate that it is a better candidate as an analgesic agent. Vomifoliol (97) is a potent binder (-4.9 kcal/mol) to COX-2 than indomethacin (-1.1 kcal/mol) indicating that it is supposed to have better analgesic action.Item The in vitro and in vivo Pharmaceutical equivalence and stability studies of some antihypertensive drugs manufactured in Bangladesh in rat model(©University of Dhaka, 2023-02-09) Begum, RehanaBackground Bangladesh is a densely populated country. To meet the healthcare needs of this huge population, huge amounts of medicines are required. Again, the first objective of National Drug Policy 2005 was to ensure that common people of Bangladesh should have easy access to effective, safe and good quality drug products at affordable prices. As hypertension is a very common disorder in Bangladesh, many pharmaceutical companies are now producing antihypertensive drugs from each class. But most of the companies do not conduct bioequivalence studies and for clinical trial and bioequivalence studies, even now we depend on another country like Malaysia, India and. No data are available in regard to pharmaceutical equivalence and bioequivalence studies of antihypertensive drugs manufactured in Bangladesh. The present study is carried out to perform in vitro and in vivo pharmaceutical equivalence and stability studies in comparison with reference innovator brands of some antihypertensive drugs manufactured in Bangladesh to compare the quality, efficacy and safety of these drug products by taking reference innovator brands as standard brands. This study will also help the physicians to choice a suitable brand which is easily available, have standards of quality, efficacy and safety. Methods In vitro pharmaceutical equivalence of some antihypertensive drugs was determined by comparing general quality assessment parameters such as weight variation, hardness, % friability, disintegration time, dissolution time and the amount of active substance between test brands and their respective reference innovator brands. Times required for 50% dissolution (T50%) and 90% dissolution (T90%) were also compared between test brands and their respective reference innovator brands. Mean of % dissolution versus time graph and statistical difference factor (f1) and similarity factor (f2) were also compared using dissolution profiles of test brands and their respective reference innovator brands. In vivo pharmaceutical equivalence of some antihypertensive drugs was done by plotting plasma concentration- time curves of test brands with their respective reference innovator brands after administration of drug in rat models. Stability testing was compared between test brands and their respective reference innovator brands under stress conditions in acidic and basic conditions at different temperatures (290C, 600C and 700C). Results Experimental three brands of tablet atenolol 50 mg were randomly designated as AA, AB, AC and reference innovator brand as ARI. Eight brands of tablet Carvedilol 6.25 mg were randomly designated as CA, CB, CC, CD, CE, CF, CG, CH and reference innovator brand as CRI. Ten brands of tablet Losartan potassium 50 mg were randomly designated as LA, LB, LC, LD, LE, LF, LG, LH, LI, LJ and reference innovator brand as LRI. Four brands of tablet ramipril 5 mg were randomly designated as RA, RB, RC, RD and reference innovator brand as RRI. All test brands including their respective reference innovator brands passed the general quality assessment parameters such as weight variation, hardness, % friability, disintegration time, dissolution and % potency. Still significant variations were observed in disintegration time of test brands of tablet losartan potassium and tablet ramipril with their respective reference innovator brands. A correlation was observed between disintegration time and the rate of dissolution in this study. All test brands including their respective innovator brands were found within % weight variation test acceptance limit. Test brands of atenolol showed weight variation percentage limit between - 2.39% and + 2.77%, whereas reference innovator brand showed weight variation percentage limit between - 1.84% and + 1.40%. Test brands of carvedilol showed weight variation percentage limit between - 2.72% and + 5.87%, whereas reference innovator brand showed weight variation percentage limit between - 1.08% and + 0.92%. Test brands of losartan potassium showed weight variation percentage limit between - 4.66% and + 4.08%, whereas reference innovator brand showed weight variation percentage limit between -2.60% and +2.14%. Test brands of ramipril showed weight variation percentage limit between -2.00% and + 2.85%, whereas reference innovator brand showed weight variation percentage limit between -1.85% and +1.49%. All test brands of tablet atenolol, tablet carvedilol, tablet losartan potassium and tablet ramipril including their respective reference innovator brands were found satisfactory for hardness testing. Test brands of atenolol showed lowest hardness value hardness between 4.55 kg and 6.13 kg, whereas reference innovator brand showed hardness 5.32 kg. Hardness of test brands of carvedilol were found between 3.88 kg and 7.68 kg, whereas 6.26 kg was found for reference innovator brand. Hardness of test brands of losartan potassium were found between 6.28 kg and 9.96 kg, whereas 6.89 kg was found for reference innovator brand. Test brands of ramipril showed hardness between 7.51 and 13.19, whereas reference innovator brand showed 7.16 kg All test brands of tablet atenolol, tablet carvedilol, tablet losartan potassium and tablet ramipril including their respective reference innovator brands met the acceptance criteria for % friability test. They had % friability values less than 1%. All test brands of tablet atenolol, tablet carvedilol, losartan potassium and tablet ramipril including their respective reference innovator brands met the acceptance criteria for disintegration time. No major variations were found in disintegration time of different test brands of atenolol. They were found to disintegrate between 0.43 and 1.36 minutes, whereas reference innovator brand disintegrated in 1.44 minutes. No momentous variations were found in disintegration time of test brands of carvedilol. They disintegrated between 0.39 and 5.33 minutes, whereas innovator brand disintegrated in 0.78 minutes. Test brands with higher disintegration time were CE, CH and CG. Significant variations were found in disintegration time of test brands of losartan potassium. They were found to disintegrate between 6.52 and 15.22 minutes, whereas reference innovator brand disintegrated in 7.19 minutes. Test brands with higher disintegration time were LA, LC, LF, LH and LI, having values ˃10 minutes. Test brands of ramipril showed significant variations in disintegration time. All test brands of ramipril disintegrated between 0.71 and 10.90 minutes, whereas innovator brand disintegrated in 1.09 minutes. Test brands with higher disintegration time were RA, RB having values ˃5 minutes and RC ˃10 minutes. All test brands of tablet atenolol, tablet carvedilol, tablet losartan potassium and tablet ramipril including their respective reference innovator brands met the acceptance limit for % of dissolution. Test brands of tablet atenolol including their respective reference innovator brand attained more than 90% of dissolution within 30 minutes. Test brands of tablet carvedilol including their respective reference innovator brand achieved more than 90% of dissolution except brand CH which got more than 80% within 30 minutes. Test brands of tablet losartan potassium LA, LC, LE, LF, LG, LH and reference innovator brand LRI did more than 90% of dissolution except brands LB, LD, LI and LJ which got more than 80% within 30 minutes. Test brands of tablet ramipril including their respective reference innovator brand attained about 100% of dissolution within 30 minutes. All test brands of tablet atenolol, tablet carvedilol, tablet losartan potassium and tablet ramipril including their respective reference innovator brands met the acceptance limit for assay content. They had % potency between 99% and 103%. The mean % of drug dissolved of tablets of different test brands were compared with that of their respective innovator brands graphically by plotting the mean % of drug dissolved against time. All test brands including reference innovator of tablet atenolol released more than 80% of drug within 10 minutes. Except test brands CB, CC, CG, CH; all other brands including reference innovator brand of tablet carvedilol released more than 80% of drug within 20 minutes. Reference innovator brand and brands LC, LE, LF, and LG of tablet losartan potassium released more than 80% of drug in 20 minutes. Test brands LA, LB, LD, LH, LI, and LJ released more than 80% of drug in 30 minutes. Except brand RC; all test brands and reference innovator brand of tablet ramipril 5mg released more than 80% of drug in 10 minutes. The time required for 50% dissolution (T50%) and 90% dissolution (T90%) were determined. All test brands of tablet atenolol and also tablet ramipril including their reference innovator brands showed T50% values less than 10 minutes and T90% values less than 30 minutes. For tablet carvedilol; all test brands including reference innovator brand showed T50% values less than 10 minutes and T90% values less than 30 minutes except test brand CH. Test brand CH had T50% less than 10 minutes but T90% greater than 30 minutes. For tablet losartan potassium; test brands LA, LB, LD, LH, LI, LJ showed T50% values greater than 10 minutes whereas, other brands less than 10 minutes. Test brands LB, LD, LI showed T90% values greater than 30 minutes whereas, other brands less than 30 minutes. The mean percentage of drug dissolved of tablets of test brands and their respective reference innovator brands were used to calculate difference factor(f1) and similarity factor (f2) using the respective equations. All test brands of antihypertensive drugs showing f1 values less than 15 are acceptable in comparison with reference innovator brands. For test brands of tablet carvedilol CB and CH; f2 values were less than 50. For test brands of tablet losartan potassium LB, LD and LI; f2 values were less than 50. For test brand of tablet ramipril RC; f2 values were less than 50. Test brands with f2 values less 50 may not be equivalent to their respective reference innovator brands. In vitro dissolution profiles showed variations in availability of drug substances from test brands and reference innovators brands. All test brands of tablet atenolol; all test brands of tablet carvedilol except two brands CB & CH; all test brands of tablet losartan potassium except brands LB, LD & LI and all test brands of tablet ramipril except brand RC were observed to have T50% values less than 10 minutes, T90% values less than 30 minutes, f1 values less than 15 and f2 values more than 50. They appeared to have very good bioavailability. Test brands CB and RC showing f2 values less than 50 but T50% values less than 10 minutes, T90% values less than 30 minutes and f1 values less than 15 also seemed to have very good bioavailability. Test brands CH and LB, LD, LI having T50% values greater than 10 minutes, T90% values greater than 30 minutes and f2 values less than 50 were not equivalent to reference innovator brands in availability of drug substances. In vivo pharmaceutical equivalence study was done by plotting plasma concentration- time curves of test brands with their respective reference innovator brands after administration of drug products in rat models. The curves indicated that the tmax value for test brands and innovator brand of atenolol was 2.5 hrs and Cmax values for brands AA, AB, AC, ARI were 0.123, 0.128, 0.113, 0.129 μg/mL respectively. The tmax value for test brands and innovator brand of carvedilol was 1.5 hrs and Cmax values for brands CA, CB, CC, CD, CE, CF, CG, CH, CRI were 0.106, 0.106, 0.102, 0.103, 0.099, 0.096, 0.096, 0.098, 0.090, 0.106 μg/mL, respectively. The tmax value for test brands and innovator brand of losartan potassium was 1.5 hrs and Cmax values for brands LA, LB, LC, LD, LE, LF, LG, LH, LI, LJ, LRI were 0.122, 0.123, 0.126, 0.118, 0.122, 0.123, 0.118, 0.123, 0.117, 0.120, 0.124 μg/mL, respectively. The tmax value for test brands and innovator brand of ramipril was 2.5 hrs and Cmax values for brands RA, RB, RC, RD, RRI were 0.047, 0.061, 0.058, 0.053, 0.063 μg/mL, respectively. Comparing in vivo Cmax and tmax values of test brands with their respective innovators, all antihypertensive testing brands may be considered equivalent to their respective reference innovator brands. Stability studies of test brands of tablet atenolol, tablet carvedilol, tablet losartan potassium and tablet ramipril including their respective innovator brands were done by stress degradation in acidic and basic conditions at different temperatures (290C, 600C and 700C). Data showed no significant degradation of test brands and also their respective reference innovator brands. So, all antihypertensive test brands also may be considered equivalent to respective reference innovator brands regarding stability. In conclusion, this study indicated that except test brands CH, LB, LD and LI; all other test brands may be considered in vitro and in vivo pharmaceutically equivalent to their respective reference innovator brands and also equivalent in case of stability. These brands may be similar in quality, efficacy, safety and may be used interchangeably. But test brands CH, LB, LD and LI are not similar to their respective reference innovator brands and cannot be used interchangeably.Item Study on Health and Drug Policies of Bangladesh to Ensure Health for All(©University of Dhaka, 2022-04-13) Rahman, Md. AknurBackground National Health and Drug policy are the standard guidelines to ensure the healthy life of the citizens of a country. According to World Health Organization, “Health is the physical, social and mental wellbeing and not merely absence of any disease”. Health is one of the basic needs of people in Bangladesh and improvement of nutritional and public health status is a Constitutional commitment of the Government of Bangladesh according to the Constitution, Article 15 and 18, respectively. The present study is focusing on the critical analysis of National Health Policy (NHP), perceptions of the service providers and service receivers regarding NHP, and Primary Health Care (PHC) in Netrokona Sadar Upazila - A Case Study on Expanded Program on Immunization (EPI), National Drug Policy (NDP), availability, affordability and price variation of essential antibiotics in Bangladesh. All issues, in the purview of NHP and NDP, have been analyzed to have an endeavor in contributing to ensure health for all. The objectives of the study were-(i) to check the consciousness of people in health sector about NHP and NDP, analyze the national budget for research and education on health and drugs, contribute on improvement of the NHP and NDP by taking some valuable suggestions from the respondents, find out the drawbacks that are in the policies and find out the strategies to deal with them; (ii) to find the areas of cooperation between Government Organizations (GOs) and Non-Government Organizations (NGOs) in the arena of PHC focusing the EPI and find the weaknesses in providing the PHC services; (iii) to measure prices of essential antibiotics for treating prevalent conditions in Bangladesh, assess the affordability of standard treatment regimens using these medicines, compare the prices of medicines found in the country with international reference prices and compare the prices of medicines of different years with statistical significance testing and (iv) to inform the policy makers to address those issues. Methods The study was conducted as survey as well as case study. Both primary and secondary sources of data were used in the study. NHP, NDP, EPI and availability, affordability and price variation of essential antibiotics of Bangladesh were analyzed. A total of 1100 respondents were selected for the study. The data were collected by random sampling method. Results NHP was consulted with the doctors, health personnel and researchers and important feedbacks were obtained. Majority of the respondents informed that they were acquainted with NHP and put some suggestions regarding the appropriateness of NHP. Role of public servant was addressed by the respondents. Some areas were identified which were not properly addressed in NHP such as management of diseases caused by Zika virus, Nipah virus, Dengue and Chikungunya, non-communicable disease, epidemic disease control. Some suggestions were also given by the respondents. Those were - need revision in NHP; update of NHP at a regular basis; involve civil society, and public representatives; ensure more punishment in case of false drug manufacture. The study showed that most of the service receivers did not know about NHP. It was observed that the percentage of people who knew about NHP was increasing with respect to the educational level. If the people are well informed about NHP, they can get better health service. The respondents put some important comments- the NHP does not ensure enough number of doctors, pharmacists (virtually absent / non-existent), nurses, and other stuffs; some medicine especially antibiotics should be prescribed by the doctors only; the NHP cannot resolve the communication gap between the service providers and the service receivers; does not ensure enough necessary supplies; does not ensure skilled doctors, pharmacists and the health workers in primary and secondary level hospitals; does not utilize the referral system properly etc. In case of service providers, acquaintance with NHP was not satisfactory. In fact, the scenario for service providers was worse than service receivers. In both the cases, it did not cross 50% and most of the service providers were not aware about the policy and as a result, they could not provide better treatment to the patients. It was evident from the study that diseases like child tuberculosis, diphtheria, whooping cough, tetanus, hepatitis B virus, Hib V, polio and measles can be prevented by vaccines through EPI as part of the endeavors of primary health care services of the government. In the journey of primary health care services through EPI, collaboration of GOs and NGOs works effectively to a great extent. The study also found some areas to be improved such as more coordination and endorsement of contributions of the players in the smooth running of the EPI. Maximum respondents (99%) opined that right use of drug practice can prevent abuse of antibiotics. Awareness and training needs were soughed in this respect. It was found that some of the respondents (22%) did not know about essential drug program. It was found that only 26 (5%) respondents out of 500 knew the exact number of essential drugs in Bangladesh. In surveyed pharmacies, the essential medicines were less available in general medicine stores as compared to the Model pharmacies. A comparative study on price variation across years had shown a insignificant increase in price. The median prices of surveyed medicines were obtained, and a gross comparison was done that indicated percent increase or decrease in price. In the studied programs, the median price ratios of surveyed medicines varied from 0.36% - 2.56% and 0.33% - 2.39% in year 2015 and 2019, respectively. While noting the WHO target that consumer should pay no more than four times the IRPs, we observed that medicine prices were lower in Bangladesh compared to IRPs. The study confirmed that the essential antibiotics were affordable in Bangladesh. Conclusion Based on our findings, it can be inferred that the idea of ensuring health for all is not a very difficult task. The major plus point has been the positive gesture of all the related stakeholders of health and medication. Contribution and cooperation of all - politicians, GOs, NGOs, civil society etc. to the NHP and NDP can make them fullest and confident to ensure health for all in Bangladesh.