Doxorubicin is one of the most widely used chemotherapy drugs in the treatment of various cancers. Despite its high efficacy, it is associated with serious side effects, including toxicity to the reproductive system. The main mechanism of these damages is attributed to increased oxidative stress, decreased antioxidant capacity, tissue destruction, and disruption of reproductive processes, including spermatogenesis in males. On the other hand, the use of plant compounds with antioxidant properties can be considered as an effective strategy to reduce these side effects. Echinacea purpurea is a medicinal plant rich in polysaccharides, flavonoids, and phenolic compounds, and its protective role against oxidative stress has been reported in numerous studies. The present study was designed to investigate the effect of hydroalcoholic extract of Echinacea purpurea on doxorubicin-induced testicular toxicity in NMRI male mice. In this experimental study, 28 male mice with an average weight of 25 to 30 grams were randomly divided into four groups of seven: the control group that received saline, the Echinacea extract group (100 mg/kg intraperitoneally every other day), the doxorubicin group (3 mg/kg intraperitoneally once a week for four weeks), and the combination group that simultaneously received doxorubicin and Echinacea extract by the above methods. After 28 days, the animals were anesthetized and blood and tissue sampling was performed. The indicators studied included testosterone levels, superoxide dismutase (SOD) activity, malondialdehyde (MDA) concentration, spermatogenesis parameters, and testicular histological changes. The obtained data were analyzed using one-way ANOVA and Tukey's post hoc test. The results of this study showed that the use of doxorubicin significantly reduced the percentage of sperm with progressive movement and viability, decreased the level of testosterone hormone and the activity of the antioxidant enzyme superoxide dismutase and increased the percentage of non-motile and non-progressive movement sperm, as well as increased the level of malondialdehyde in the serum of mice (p?0.05). The use of Echinacea extract alone improved the parameters of sperm motility and viability, increased testosterone levels, increased SOD activity and decreased MDA (p?0.05). In the group receiving Echinacea extract and doxorubicin simultaneously, the negative effects of doxorubicin on sperm motility and viability and oxidative stress indices were slightly modified, so that the percentage of live and non-motile sperm and MDA levels improved compared to the doxorubicin group, although the increase in SOD activity was not significant. Histological examination of the testes showed that doxorubicin caused structural damage, but concomitant administration of Echinacea extract was able to significantly modulate this damage, indicating the protective effect of Echinacea extract against doxorubicin toxicity on testicular tissue. Overall, the findings of this study indicate that Echinacea hydroalcoholic extract, with its antioxidant properties, is able to modulate part of the damage caused by doxorubicin on the male reproductive system and can be considered as a potential complementary option to reduce the side effects of chemotherapy.

      طب سنتي يكي از علوم تجربي است كه با ظهور و پيدايش علوم مدرن همچنان اهميت و جايگاه خود را مانند گذشته حفظ كرده است. طب سنتي كه در گذشته حاصل كسب تجارب در زمينه­ي استفاده از روش­هاي درماني مختلف بوده است امروزه به صورت آكادميك و سازمان يافته شده تدريس و مورد بهره برداري قرار مي­گيرد. طب سنتي بنابر تعريف ارائه شده توسط سازمان بهداشت جهاني عبارت است از مجموعه­اي از مهارت­ها، دانش و روش­هاي مبتني بر تجربه كه به منظور پيشگيري، كاهش و درمان انواع بيماري­ها مورد استفاده قرار مي­گيرد. كتاب الصيدنه في الطب (داروشناسي در پزشكي) آخرين اثر ابوريحان بيروني است كه به صورت تنها نسخه­ي اصلاح نشده به زبان عربي به ما رسيده ‌است. اين اثر گرانبها منبعي در داروشناسي سده‌هاي ميانه­ي خاورزمين است كه در آن بيش از هزار دارو با منشا گياهي، حيواني و معدني با اشاره به نام‌هاي آن‌ها به بسياري از زبان‌ها و گويش‌ها توصيف شده‌ است. در صيدنه اثري از مدح و اهدا نامه معمول ديده نمي‌شود و از هدف‌هاي نوشتن آن نيز مستقيما سخني نرفته ‌است. بيروني با درك اهميت زياد اين مسئله، حدود ???? نام گياه، حيوان، ماده­ي معدني و فراورده‌هاي آن‌ها را به زبان‌هاي عربي، يوناني، سرياني، هندي، فارسي، خوارزمي، سغدي، تركي و جز اين‌ها گرد آورده و توضيح داده ‌است و بدين ترتيب گام جديدي در راه تنظيم و ترتيب اصطلاحات دارويي زمان خود برداشته ‌است. با توجه به اين كه تا كنون يك بررسي جامع در خصوص اين كتاب انجام نشده است اين پژوهش انجام شد و اميدواريم كه در آينده بتوانيم اطلاعات موجود در اين كتاب ارزشمند را استخراج و در صورت امكان به صورت يك پايگاه داده­ي مناسب درآوريم.    واژگان كليدي: طب سنتي، الصيدنه في الطب، پايگاه داده.       طب سنتي يكي از علوم تجربي است كه با ظهور و پيدايش علوم مدرن همچنان اهميت و جايگاه خود را مانند گذشته حفظ كرده است. طب سنتي كه در گذشته حاصل كسب تجارب در زمينه­ي استفاده از روش­هاي درماني مختلف بوده است امروزه به صورت آكادميك و سازمان يافته شده تدريس و مورد بهره برداري قرار مي­گيرد. طب سنتي بنابر تعريف ارائه شده توسط سازمان بهداشت جهاني عبارت است از مجموعه­اي از مهارت­ها، دانش و روش­هاي مبتني بر تجربه كه به منظور پيشگيري، كاهش و درمان انواع بيماري­ها مورد استفاده قرار مي­گيرد. كتاب الصيدنه في الطب (داروشناسي در پزشكي) آخرين اثر ابوريحان بيروني است كه به صورت تنها نسخه­ي اصلاح نشده به زبان عربي به ما رسيده ‌است. اين اثر گرانبها منبعي در داروشناسي سده‌هاي ميانه­ي خاورزمين است كه در آن بيش از هزار دارو با منشا گياهي، حيواني و معدني با اشاره به نام‌هاي آن‌ها به بسياري از زبان‌ها و گويش‌ها توصيف شده‌ است. در صيدنه اثري از مدح و اهدا نامه معمول ديده نمي‌شود و از هدف‌هاي نوشتن آن نيز مستقيما سخني نرفته ‌است. بيروني با درك اهميت زياد اين مسئله، حدود ???? نام گياه، حيوان، ماده­ي معدني و فراورده‌هاي آن‌ها را به زبان‌هاي عربي، يوناني، سرياني، هندي، فارسي، خوارزمي، سغدي، تركي و جز اين‌ها گرد آورده و توضيح داده ‌است و بدين ترتيب گام جديدي در راه تنظيم و ترتيب اصطلاحات دارويي زمان خود برداشته ‌است. با توجه به اين كه تا كنون يك بررسي جامع در خصوص اين كتاب انجام نشده است اين پژوهش انجام شد و اميدواريم كه در آينده بتوانيم اطلاعات موجود در اين كتاب ارزشمند را استخراج و در صورت امكان به صورت يك پايگاه داده­ي مناسب درآوريم.    واژگان كليدي: طب سنتي، الصيدنه في الطب، پايگاه داده.   

   In the tumor microenvironment, differences in cancer cells' access to nutrients and oxygen modify cellular metabolism. Hypoxic cancer cells turn to glycolytic metabolism to survive and proliferate, producing large amounts of lactate that must be tra  orted out of the cell. During a metabolic symbiosis, oxidative cancer cells import the excess lactate and use it as a preferred fuel instead of glucose. Lactate tra  ort is facilitated by monocarboxylate membrane tra  orters (MCTs) belonging to the solute carrier gene family-16, which is a proton-dependent process and plays a role in intracellular pH regulation. The export of lactate from the cell is mainly facilitated by MCT4, while MCT1 mediates its intracellular uptake. Overexpression of these tra  orters has been shown to be associated with a variety of malignancies, including breast, gastric, lymphoma, brain, lung, skin, and soft tissue cancers, and targeting them could be a potential treatment for some types of cancer. In this study, we used various virtual screening techniques including pharmacophore modeling, molecular docking and molecular dynamics simulation to identify effective and potential drug candidates against human MCT1. The atomic coordinates of the protein in outward-open conformation were downloaded from the protein data bank with the code 6LYY, and a library of chemical compounds including 12 million molecules purchasable from the ZINC database and 4683 FDA-approved drugs was created. After performing the preparation steps, molecular docking calculations were performed based on a consensus approach using AutoDock Vina, Molegro, and DOCK programs. Ligands with high binding energy were analyzed in successive steps, and their interaction with key residues of the protein active site was investigated. Finally, seven ligands that showed promising results were selected for the molecular dynamics simulation study. For each protein-ligand complex in the membrane bilayer, calculations of protein backbone RMSD, ligand RMSD, RMSF, and interaction analyses were performed. The results showed that Olmesartan, an angiotensin II receptor inhibitor, can have an inhibitory effect on human MCT1 with a strong and stable binding and pave the way to inhibit this tra  orter. Since this study is based solely on computational tools, further evaluation in experimental conditions is necessary to confirm its effectiveness.

   Natural products, including herbal formulas and extracts, have been used to treat human diseases for thousands of years, serving as valuable resources for treating diabetes. This study aimed to investigate the simultaneous effect of eugenol and ascorbic acid on the expression of genes involved in the regulation and tra  ort of glucose in diabetic rats treated with alloxan. To carry out this study, first after preparing the rats, keeping them and adapting them to the environmental conditions, the target groups were made diabetic by injecting 180 mg/kg body weight of the chemical substance alloxan and after making sure that they had diabetes; treatment with gavage of 10 mg/kg of body weight of eugenol and injection of 100 mg/kg of body weight of ascorbic acid to the target groups continued for 45 days. In the next step, the rats were anaesthetized, and after dissection, their liver and pancreas tissues were separated. After quick freezing with liquid nitrogen, they were placed in a -80 degree refrigerator. In the continuation of the work, RNA was isolated from all liver and pancreas tissues and cDNA synthesis was performed for all treatment and control samples. Then the expression of genes in pancreatic and liver tissue was checked using the Real-time PCR method. The results showed that the expression of Ins1/2, InsR, Glut2, and Pdx1 genes in the pancreatic tissue of treated rats increased significantly (P ? 0.05) compared to the diabetic control group, and the expression of Tnf? gene in the pancreatic tissue compared to the control group Diabetic showed a significant decrease in expression. In addition, in the liver tissue of the treated group, the expression of Glut1 and Glut2 genes significantly decreased compared to the diabetic and non-diabetic control groups, the expression of Tnf? gene compared to the diabetic control group without significant change, and increased compared to the non-diabetic control group, the expression of InsR gene showed no significant change compared to the diabetic control group and a significant decrease compared to the non-diabetic control group. Therefore, investigating the simultaneous effect of eugenol and ascorbic acid on the expression of genes involved in the regulation and tra  ort of glucose in diabetic rats showed that these natural products have an effect on blood sugar levels and also the antidiabetic capacity of the samples Treated with these products compared to tissue samples of diabetic control, they showed a significant increase in expression and a decrease in expression compared to non-diabetic control. Keywords: eugenol, ascorbic acid, rats, diabetes, glucose tra  ort

Abstract    Today, infertility is considered one of the most important problems in men, in such a way that clinical and epidemiological evidence has confirmed the increase in infertility in men. Several factors such as ionizing rays, magnetic fields, smoking, and some drugs lead to an increase in infertility in men. On the other hand, due to the effectiveness of some herbal medicines and chemical compounds in increasing male fertility, extensive research has been done to discover biologically active substances that can overcome the problem of male infertility. Also, due to the wide applications of stem cells, especially spermatogonial sperm cells (SSCs), and the use of natural substances, it has provided new hope for the treatment of many human diseases, especially infertility and maintaining fertility in men. vitamin C, due to the glutathione enzyme dependent on dehydroascorbic acid reductase, keeps the amount of vitamin C in the testicular tissue at a high level, in increasing the activity of sperm movement, increasing the quality Semen and fertility of rats play an important role, also Eugenol is a phenolic compound derived from clove extract, which has antioxidant, anti-inflammatory, and anti-cancer effects, Few studies have been done on the effect of these two compounds on SSCs. Therefore, this study was designed with the aim of studying the effect of vitamin C and eugenol treatment on fertility and the expression of genes involved in self-renewal (Plzf, Sox2) and differentiation (Dazl) of spermatogenic stem cells in rats. For this purpose, after treating the animals, their testicular tissue was separated. The expression patterns of different molecular markers in the testis were investigated using RT-PCR methods and histomorphological analyses of the testis tissue and the counting of its spermatogonia, spermatocytes, and spermatids.The results of this study showed an increase in the number of spermatogonial, spermatocytes, and spermatids cells in testicular tissue. In addition, it was found that the genes involved in self-renewal (Plzf, Sox2) and differentiation (Dazl) are expressed at the transcript level. Also, this study indicated that the use of eugenol alone had a greater effect than its treatment with other compounds, in addition, it was determined that vitamin C in the body according to the environmental conditions of the cell and the presence of free transition metals such as copper and iron. In addition to its antioxidant properties, it can also have a pro-oxidant effect.Overal, this study has presented a new perspective on the effect of natural substances and antioxidants on the expression pattern of molecular markers in SSCs and the increase in the number of sperm progenitor cells in rat testicular tissue.  Keywords: Treatment, Vitamin C, Eugenol, Self-renewality, Differentiation, Spermatogonial stem cells, Rat, Fertility  

  Carbonic anhydrase (CAs, EC 4. 2. 1. 1) is a family of zinc metalloenzymes that catalyzes vital reactions including the reversible conversion of carbon dioxide and water to bicarbonate and a proton. Carbonic anhydrase isozymes are involved in various physiological and pathological processes and are considered as important drug targets for the treatment of a wide range of disorders including glaucoma and various types of cancer. In this study, new sulfonamide derivatives resulting from the coupling reaction of sulfanilamide with benzene or its mono-, di-, tri- carboxylic acid compounds were chemically synthetized and their interaction with hCA II isozyme were investigated by various spectroscopic techniques. Kinetic results revealed that new sulfonamide derivatives inhibit the esterase activity of hCA II in a reversible competitive manner. As a result, among the studied compounds, compound 4 had the lowest Ki and IC50 values for hCA II isozyme. Also, fluorescence measurements showed that these compounds quench the intrinsic fluorescence of hCA II by a dynamic quenching mechanism. In addition, analysis of the thermodynamic parameters of the binding revealed that hydrogen bonds and van der Waals interactions play the major role in stabilization of the enzyme–drug complexes. Fluorescence analysis of the CAII-DNSA fluorescent complex in the presence of different concentrations of new sulfonamide derivatives showed the lowest dissociation constant (Kd) for the compound 4, indicating a higher affinity of this compound for the binding to the hCA II isozyme. Overall, the strengthening of the binding power and inhibitory activity of the studied sulfonamide derivatives for the hCA II isozyme, makes these derivatives of great interest for the design of novel hCA inhibitors.

Over the past three decades, the increase in the number of people with diabetes has made it one of the most important challenges to all nations. The global prevalence of diabetes mellitus is increasing rapidly following lifestyle changes. Type2 diabetes mellitus is a complex multifactorial disease characterized by the interaction between multiple genetic loci and various environmental factors. Differences in the risk of developing type2 diabetes between different ethnic groups indicate the influence of genetic predisposition to developing it. The OPRM1 gene encodes the mu-opioid receptor, which is the major target for both endogenous and exogenous opioids in the pain relief process. Several studies examining the association between type2 diabetes and the 6q24-q27 region have suggested a role for the OPRM1 gene, based on its chromosomal location, in the risk of developing the disease. Another research suggests that the mu-opioid receptor may modulate genes involved in glucose metabolism, leading to lower plasma glucose levels. The most common single nucleotide polymorphism (  ) in the coding region of the human OPRM1 gene is the A118G variant, which results from the replacement of adenine with a guanine. This variant has been reported to alter both endogenous ligand (?-endorphin) binding and receptor signalling following this binding. The aim of this study was to investigate the association of A118G polymorphism in the OPRM1 gene with type2 diabetes mellitus because according to the above, the study of the gene polymorphisms can provide a new strategy for the prognosis of the disease and improvement of treatment strategies. In this study, 207 diabetic and 201 non-diabetic individuals (as controls) were selected and blood samples were taken from these individuals. Blood samples were used to extract DNA. The extracted DNA was used for PCR reaction to analyze for polymorphism using primers pre-designed by Primer1 online software, and then the PCR product was used for sequencing studies. Finally, the results of sequencing were analyzed to examine the presence of the    in different individuals. Sequencing results showed the presence of the corresponding polymorphism in the diabetic samples, while it was not found in control samples. Therefore, given the role of the OPRM1 gene in glucose metabolism, the presence of this polymorphism may be related to the risk of developing type2 diabetes; However, definitive results require further research.   

  Arginine is a semi-essential amino acid, which participates in many metabolic pathways, such as nitric oxide, creatine and urea synthesis. The availability of arginine is determined by a series of plasma membrane carries called cationic amino acid tra  orters (CATs), which are overexpressed in many types of cancers including Acute myeloid leukemia (AML) that is one of the most common cancers in adults and children; therefore, these tra  orters have been considered as a putative target in therapeutic and medicinal goals. In the present research, the mechanism of L-arginine tra  ort through CAT1 and hotspot amino acids in the tra  ortation was investigated using molecular dynamic simulation techniques. For this purpose, prokaryotic CAT1 (p-CAT) with PDB ID 6F34 was received from Protein Data Bank (PDB). Since eukaryotic CAT1 (e-CAT) has not yet solved, the e-CAT structure was modeled using MODELLER 9.20 based on p-CAT as template. After that, because the CAT proteins are membrane tra  orters, the proteins were inserted in the suitable membranes using CHARMM-GUI server. Then, molecular dynamics simulation was performed for 300 nanosecond (ns) for each system. In the next stage, L-arginine (L-Arg) was docked against both the equilibrated system using AutoDock VINA, and molecular dynamics simulation was run for 600 ns for each system. In order to calculate total free binding energy of L-Arg, MM-  A approach was carried out on both p-CAT and e-CAT trajectories. To determine the contribution of each amino acid residue to the binding energy, the per-residue energy analysis was performed. In the p-CAT/Arg complex, Ile40, Thr43, Asp111, Glu115, Lys191, Phe230, Ile234 and Asp237 had the greatest contribution toward the binding to L-Arg and were formed stable hydrogen bonds with it. The residues Ser44, Glu185, Asp270, Cys271, Ser350, Ser354 and Arg360 were found to be the highest contributing residues that interact with L-Arg in e-CAT/Arg complex. In order to evaluate the tra  ortation processes through CAT tra  orters and also to study more precisely the role of hot spot amino acids during tra  ort, the umbrella sampling method was employed. Hydrogen bond analyses revealed that in p-CAT, the residues Asp237, Glu115, Ser321, Glu245, Lys19 are the most involving residues in interaction with L-Arg during tra  ortation. In e-CAT, the residues Asp270, Glu185, Ser22, Asp20 and Arg27 are the most involving residues in interaction with L-Arg during tra  ortation. To investigate the effects of membrane lipids on the structures of p-CAT and e-CAT, membrane analyzes such as lipid-protein interaction, MSD, SCD, density and membrane thickness were performed for both systems. The results showed that protein loops had reduced flexibility due to interaction with lipids. Further, in eukaryotic membrane, due to the ordering effect of cholestrol, the SCD value and membrane thickness are higher than prokaryotic membrane.Key words: L-Arginine, CAT1 protein, molecular dynamics simulation, umbrella sampling

  Human serum albumin nanoparticles as an effective exemestane drug delivery system to treat breast cancer: preparation and analysis

   خانواده سوسماران مانيتور يا   Varanidae يكي از نه خانواده سوسمار­هاي ايران است كه تنها داراي يك جنس (مونوفايلتيك) است اما حدود   73گونه در اين جنس قرار دارد. مانيتور­ها يا بزمچه­ها بزرگترين سوسماران جهان از لحاظ جثه هستند بزمچه­ها داراي بدن كشيده، پوزه بلند، كشيده و دوكي شكل هستند و دم در آنها بلند و حدود 5/1 برابر طول بدن است، زبان بلند و دو شاخه و چشم­ها با پلك متحرك و مردمكي گرد است، بزمچه­ها روز گرد و شكارچي بوده و آرواره بسيار نيرومندي دارند از جنس وارانوس گونه­هاي الف) بزمچه بنگال (1802، Daudin) Varanus Bengalensis ب) بزمچه بياباني (1803، Daudin) Varanus griseus ج) بزمچه نسترو Varanus nesterovi )(B?hme, Ehrlich, Milto Orlov & Scholz, 2015)   در ايران مشاهده شده است. مطالعه روي جمجمه سوسماران بر روي مسائل مربوط به اختلافات بين جمجمه در موجودات متنوع و منشاء آنها و علت اين اختلافات، تمركز كرده است. در ايران مطالعه جامعي در مورد اسكلت كامل سوسماران صورت نگرفته، و اين در حالي است كه اسكلت تاثير بسزايي در زندگي، حركت، شكار، تغذيه و ... دارد و محيط نيز تاثير مستقيم بر اسكلت جاندار مي گذارد. در اين مطالعه به بررسي اسكلت ( Varanus griseus (Daudin, 1803 و B?hme, Ehrlich, Milto Orlov & Scholz, 2015 Varanus nesterovi مي­پردازيم. در اين پروژه براي بررسي اسكلت، از دو روش استفاده شده است. ?. خارج كردن اجزاء اسكلتي و عكسبرداري از آن­ها. ?. استفاده از روش CTscan. نتيجه استخراج 302 قطعه استخوان ازبدن سوسمار كه 28 قطعه متصل به هم جمجمه جانور را تشكيل مي­دهد. در مرحله بعد با استفاده از تصاوير اسكن استخوان­هاي خارج شده نامگذاري و اتصالات اسكلتي شناسايي شد. مقايسه اسكلت با گونه­هاي ديگر جنس وارانوس و خانواده­هاي ديگر سوسماران انجام شد و نتيجه وجود تفاوت اندك بين گونه­اي و اختلافات چشمگير بين خانواده­ها مي­باشد كه اين اختلاف­ها ناشي از عوامل متعدد مثل جدايي تكاملي، تفاوت در نوع محيط زيست، تغذيه و رفتار جانور است.

  Carbonic anhydrases (EC 4.2.1.1) are zinc metalloenzymes that catalyze the reversible hydration of CO2 to HCO3? and proton. These enzymes have different distribution in different tissues and subcellular locations, and found in the archaea, eubacteria, animals and plants.These enzymes contributed in vital physiological processes associated with respiration and transfer CO2, secretion of electrolytes in tissues and lungs, pH adjustment, homeostasis, biosynthetic reactions (such as gluconeogenes and lipogenes) and calcification. Carbonic anhydrase inhibitors are a 20px;">Considering all the above data, it can be concluded that binding of 1-(4-chloro-benzenesulfonyl)-4-hydroxy-pyrrolidine-2-carboxylic acid to human carbonic anhydrase II caused changes   in the function as well as in the secondary and tertiary structure of the protein. Keywords: 1-(4-chloro-benzenesulfonyl)-4-hydroxy-pyrrolidine-2-carboxylic acid, Human Carbonic Anhydrase II, Inhibition, Thermodynamic Stability, Kinetic Stability, Fluorescence, Quenching

  Carbonic anhydrases are well known zinc metalloenzymes involved in the catalysis of carbon dioxide hydration to bicarbonate and proton. The physical and chemical properties of phenolic compounds make these molecules capable of interacting with a wide range of targets, such as the Carbonic anhydrases. In this study the inhibition of two human carbonic anhydrase isozymes II and IX, with a series of phenol derivatives was investigated by using the esterase assay, with p- nitrophenyl acetate as substrate. The IC50 values of quercetin (Q) and its sulfonamide derivative (QD) were 15.99 and 5.13, for carbonic anhydrase II, 54.45 and 28.52 for carbonic anhydrase IX, respectively. These ligands can quench the intrinsic fluorescence of CAII by dynamic quenching mechanism. As the conclusion, binding of these phenolic compounds to the active site of CAII is accompanied by a competitive inhibition of the enzyme. According to the results, these phenolic compounds were more potent inhibitors for CAII Compared to CIX.

In the present investigation, we analyzed the interaction of resorcinol and resveratrol and their sulfonamide derivatives with human carbonic anhydrase II (hCA II) using fluorescence spectroscopy, molecular docking and molecular dynamics simulation techniques. Fluorescence data obtaining at three temperatures indicated that resveratrol and its sulfonamide derivative quenched intrinsic fluorescence of the enzyme through a static mechanism but resorcinol and its sulfonamide derivative increased intrinsic fluorescence of the enzyme again through a static mechanism. Thermodynamic analysis of the quenching data indicated that hydrogen bonding and van der Waals interactions play important roles in the ligand binding. Based on computational data obtained by molecular docking and molecular dynamics simulation studies, hydrogen bonds are the main intermolecular forces for the ligand-hCA II interactions. In comparison with resveratrol and resorcinol, their sulfonamide derivatives bind stronger to hCA II. According to an assay method basing on fluorometric measurements, the sulfonamide derivatives had a greater inhibitory effect than the original compounds.

Serum Albumin is the most abundant protein in the blood circulatory system. Its main duty determinant of plasma oncotic pressure. According to Crystal structure analysis of Human Serum Albumin (HSA) has revealed that it consist of a 585 amino acid residue and its orders three homologous domains (I–III) that each one divided into subdomains A and B. HSA has an extraordinary ligand-binding capacity, making a depot and carrier for many endogenous and exogenous ligands. As for capable HSA of binding to a wide variety of drugs. Thiazide-type diuretics are a first-line therapeutic for patients suffering from hypertension Hydrochlorothiazide is one of these that developed and introduced for more than 50 years. This study was designed to examine the interaction of Hydrochlorothiazide (HCTZ) as a ligand with Human Serum Albumin (HSA) under physiological conditions .To this aim, we are using different biophysical techniques such as UV-vis absorption, circular dichroism (CD), FT-IR Spectroscopy and also fluorescence spectroscopic methods to investigate the binding HCTZ.The results of fluorescence titration revealed that the HCTZ strongly quench the intrinsic fluorescence of HSA through a dynamic quenching mechanism. Binding constants ( ) and the number of binding sites were calculated by using Stern-Volmer equations. Subsequently, the values of   and   were also calculated, which revealed that the positive enthalpy and entropy values of the interaction of HCTZ and HSA indicate that the hydrophobic interactions played a major role in drug binding process. Computations of the protein surface hydrophobicity (PSH) index by using 1-anilinonaphtalene-8-sulfonate (ANS), also indicated an increased in PSH value upon drug binding. The binding site has been identified by using Warfarin and Ibuprofen as site marker. Also Molecular Docking results indicated HCTZ   is placed at subdomain IIA. The results of study far- and near-UV CD experiments showed some variations in the secondary and tertiary structures of protein upon ligation. Also the results of FTIR illustratived that the drug interacted with HSA and induced a partial compactness in the secondary structure of protein. The results identified by the FTIR are in good agreement with CD spectra.