Using nano polymeric micelles based on beta cyclodextrin for the targeted delivery of bosentan drug

Using nano polymeric micelles based on beta cyclodextrin for the targeted delivery of bosentan drug | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Using nano polymeric micelles based on beta cyclodextrin for the targeted delivery of bosentan drug Hossein Ameli, Nina Alizadeh This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4227684/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The drugs formulation have been considered based on the micelles of polymeric nanoparticles in the past decades. It has shown that these Nano polymeric micelles (Nano PMs) help to increase accessibility to homeland, decreasing side effects and getting long of drug dissemination. The purpose of this study is optimization of the bosentan drug formula using polymer nanomicelles and cyclodextrins. The pH-responsive copolymer was prepared and the variables of loading time, loading temperature, the ratio of CP5 copolymer to drug and the ratio of cyclodextrin to drug and the effect of these variables on the percentage of drug loading were investigated. In this work, first, a Definitive Screening Design was used to investigate the effect of variables on drug loading percentage. This method provides the possibility of significantly reducing the number of tests required to check systems that are characterized by a large number of variables. In the optimized formula for the drug, the loading temperature was 43 °C and the time was 4.25 hours, as well as the ratio of CP5 copolymer to the drug was 4.7 and the ratio of cyclodextrin to the drug was 2.4 to reach the maximum efficiency of 84.5% drug entrapment. The release was carried out in laboratory conditions at pH 1.2 and 7.4 and at a temperature of 37 °C for 24 hours, 95% of the drug was released, which showed that the release of the drug was completely targeted and controlled. Therefore, according to the obtained results, we can hope that polymer nanomicelles with cyclodextrin are potential and efficient carriers for targeted delivery with sustained release of drugs. Bosentan Nano polymeric micelles Cyclodextrin Definitive Screening Design Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1. Introduction Cardiovascular disease (CVD) causes almost 18 million deaths per annum. Along with strong evidence, hypertension associated with high blood pressure have a high-risk factor for CVD. Among the types of hypertension, pulmonary arterial hypertension (PAH) is perilous with 20% mortality. It results in right coronary failure and death[ 1 – 4 ]. Pulmonary arterial hypertension (PAH) is a life threatening disease that affects about 3 million people per year. It is often fatal with a mean life about three years after diagnosis [ 5 – 7 ]. Symptoms comprise shortness of breath, chest pain, syncope, fatigue and peripheral edema which is more prevalent in females than in males [ 8 ]. Patients with PAH suffer from elevated levels of endothelin (ET1); a potent blood vessel constrictor, in their plasma and lung tissues [ 9 ]. ET1 reduces pulmonary arterial lumen diameter, increases pulmonary vascular resistance, decreases reactivity of the vascular bed and eventually increases pulmonary arterial pressure [ 10 ]. Currently, endothelin receptor antagonists (ERA) are used for the treatment of PAH. Bosentan (tracleer) was the first orally active ERA to reach the clinical investigation stage [ 11 , 12 ]. Bosentan is a dual ERA with high affinity of both ETA and ETB receptors. Bosentan is indicated for the treatment of pulmonary artery hypertension (PAH) by blocking the action of endothelin molecule [ 13 – 15 ] and it can prevent or reverse the associated histological change caused by elevated levels of ET1. Bosentan and its metabolites are eliminated predominantly in the liver by the cytochrome P450 isoenzymes, CYP2C9, CYP3A4 and the terminal half-life after oral administration is 5.4 h [ 6 ]. It is available in two concentrations 62.5 mg and 125 mg (for twice daily administration) and its main side effects are headache, flushing and increased liver aminotransferases. Hepatotoxicity caused by bosentan is dose-dependent in higher dosages [ 16 , 17 ]. The poorly soluble drugs typically limit their absorption after oral administration, ultimately limiting clinical usefulness. Bosentan is a poorly soluble oral dual endothelin receptor antagonist, usually indicated for the treatment of pulmonary arterial hypertension (PAH) [ 18 ]. It shows an absolute bioavailability of about 50% with plasma elimination half-life of 5.4 h [ 19 , 20 ]. Considering its low aqueous solubility, slow dissolution rate in gastrointestinal fluids, short duration of action, frequent oral administration, and dose-dependent hepatotoxicity [ 21 ], nanomicellar -based delivery system may be a better option. Nanomicellar exhibit great promise as drug delivery carriers due to their well-documented thermodynamic stability, improved capacity of solubilization, relatively low viscosity, and steric protection against protein adsorption and increased capability of cellular uptake than the free drug [ 22 ]. The self-assembled nanomicelles are composed of a hydrophobic core surrounded by a hydrophilic shell (corona) of polymer, which provide effective steric protection and enable them to remain in dispersed state, prohibit recognition by mononuclear phagocyte system (MPS), ultimately stabilizing the nanomicellar structures. The prolonged circulation of micellar assists in maintaining the required therapeutic level of drug in the blood over an extended period [ 23 ]. The incorporation of lipophilic drugs into the core of polymeric micellar could enhance water solubility, enhanced permeability across the physiological barriers and cause substantial improvement in oral bioavailability, and therapeutic activity with consequent reduction in dose-dependent side effects associated with the drug. As drug carriers, polysaccharidebased core-shell nanomicellar systems have demonstrated great potential to improve the delivery of hydrophobic drugs through enhanced solubility, stability, and controllable drug release properties. Various hydrophobic moieties have been conjugated to the backbone of these polysaccharides to impart them amphiphilic character. These include but not limited to cholesterol [ 24 – 26 ], deoxycholic acid [ 27 , 28 ], stearic acid [ 29 ], octane or dodecane [ 30 ], oleoyl group [ 31 ], cetyl group [ 32 , 33 ], and so on. Despite a significant progress, the micellar systems are still under development to search for novel biomaterials and meet the clinical need. Cyclodextrins are naturally available water-soluble cyclic oligosaccharides-1,4-linked-glucopyranose composed of six or more glucose units [ 34 ]. Cyclodextrins are well known for their ability to increase solubility, dissolution rate and bioavailability of the loaded drugs [ 35 ]. They have a ring structure with a hydrophilic outer surface and lipophilic cavity which provide them with the ability to form non-covalent inclusion complexes with drug molecules of appropriate size. Inclusion complexes are very well explored to improve the water solubility and chemical stability of the drugs [ 36 ]. The nano polymeric micelles comprise a drug-loading core and a hydrophilic shell. Amphiphilic block copolymer forms micelles when in contact with an aqueous vehicle by self-assembly resulting in hydrophobic interactions wherein hydrophobic drugs can be encapsulated into the central core of micelles through hydrophobic interactions [ 37 – 39 ]. The presence of this hydrophilic corona makes the micelles more stable and stealthy, helping them to avoid the reticular-endothelial system, also providing prolonged circulation and residence time in the bloodstream. These advantages, together with their small size (100 nm), make the polymeric micelles promising carriers for the administration of various insoluble and poorly soluble pharmaceutics that can be incorporated into the hydrophobic core of the micelles [ 40 – 43 ]. Another advantage of the polymeric micelles is their ability to form various types of pH-responsive drug delivery systems depending on the encapsulated drug and physiological destination. The Incorporation of pH-sensitive groups into the core-forming blocks can make the polymeric micelles sensitive to the environmental pH. In the most common strategy, ionization of the pH-sensitive groups of the inner blocks converts the micelle core from hydrophobic to hydrophilic, resulting in the demicellization of the copolymers and rapid release of the encapsulated hydrophobic drug. Polymeric micelles containing acidic groups in their inner core can be used to design smart oral drug delivery systems [ 44 – 48 ]. Such assemblies are stable at the acidic pH of the stomach because their inner blocks are in their unionized and hydrophobic form. Upon a pH increase in the intestine, the acidic groups start deprotonation which increases the electrostatic charge and hydrophilicity of the inner part, leading to the micelle dissociation and drug release. The key element for the pH sensitivity of a polymer in the presence of ionizable pendant groups which attach to the hydrophobic backbone of the polymer chain. pH-sensitive polymers are, thus, a class of polyelectrolytes with ionic functional groups that are weak acidic ( e.g. , carboxylic and sulfonic acids) or basic ( e.g. , amines, imidazole, and pyridine) groups [ 49 – 53 ]. The Copolymer of Maleic and Acrylic Acid is a copolymer of maleic anhydride and acrylic acid. It does not degrade below pH 7 thus providing film coats that are resistant to gastric media but soluble in intestinal fluid. A combination of Nano PMs and β-CD can improve the drug loading capacity and therapeutic efficacy of poorly water soluble drugs. In the analysis of problems affected by several factors with possible interactions, statistical screening methods are often adopted to select the parameters that actually affect the response variable and to eliminate the irrelevant ones. This is particularly useful at the beginning of an investigation when little or no information is available for the system of interest. Recently, a new class of three-level designs, the so-called definitive screening designs (DSD), has been proposed by Jones and Nachtsheim [ 54 ]. DSD allows for assessment of active effects, two-factor interactions and pure-quadratic effects in the presence of effector sparsity. This allows a dramatic reduction in the number of experiments, thus enabling a significant saving in time and cost of materials. The use of these screening methods is increasing in recent years [ 55 – 57 ]. Thus, the objective of this present investigation was to synthesize pH-sensitive nano polymeric micelles as controlled release carriers for bosentan. 2. Materials and methods 2.1. Materials β- Cyclodextrin (purity, > 98%) was purchased from Sigma, The drug Bosentan monohydrate (purity, > 99%) was taken from Abmole Company, Acrylic/maleic copolymer (purity, > 92%) With a molar mass of 70,000 g/mol And brand CP 5 powder was purchased from BASF Germany, dialysis membrane (MWCO 12 kDa) was purchased from Sigma, and all other reagents and chemicals were of analytical grade and purchased from sigma. 2.2. Characterization The UV-Vis absorption measurements were taken at room temperature using UV 1800 spectrophotometer (SHIMADZU, Japan). The UV-Vis of the samples were recorded in the frequency range of 200 ~ 700 nm. The FTIR spectra of the Nanocomposites-bosentan nanoparticles and free Bosentan were obtained with a (protege 460) FTIR Spectrometer (Nicolet, USA). For FTIR spectroscopic investigations, 10 mg of the sample were mixed with 100 mg KBr and pressed into a pellet. The measurements were carried out in the mid-infrared range of 4000 cm − 1 to 400 cm − 1 . The resolution was set to 4 cm − 1 ; 100 scans were recorded, averaged for each spectrum. 2.3. Preparation of Bosentan loaded Nano polymeric micelles The inclusion complex of bosentan, cyclodextrin and copolymer was prepared using the evaporation method with solvents. Equivalent amounts of pure drug, cyclodextrin and copolymer were dissolved in 15 ml of methanol and 85 ml of distilled water, respectively. This solution was placed in an ultrasonic bath for 15 minutes. The solution was transferred to a conical flask and stirred using a magnetic stirrer for 4 hours at a suitable temperature at 150 rpm. The resulting solution was evaporated at a temperature of 45°C and the dried complex was passed through sieve No. 44[ 58 ]. 2.4. Determining the amount of drug and entrapment efficiency of bosentan To measure the amount of drug trapped in nano polymeric micelles (Nano polymeric micelles), we take a certain amount of it and dissolve it in methanol, because nano PMs are not dissolved in methanol and only the drug bosentan that is not trapped is dissolved in methanol. Then it is centrifuged and the concentration of the free drug in the supernatant is measured using a UV-Vis device at a wavelength of 271 nm, and as a result, the amount of trapped drug is obtained by Eqs. 1 and 2. C e = C i - C s (1) EE (entrapment efficiency) = \(\frac{\mathbf{C}\mathbf{e}}{\mathbf{C}\mathbf{i}}\times 100\) (2) Where, C e is the Amount of drug trapped (encapsulated); C i is the Initial drug value; C s is the drug value in the supernatant; EE is the drug entrapment efficiency. 2.5. Experimental design The DSD proposed by Jones and Nachtsheim [ 54 ] was adopted to investigate the effects of 4 continuous factors (k = 4) for Bos-loaded Nano polymeric micelles that were identified, in preliminary runs, as potentially important for the drug loading process. They were temperature, time, cyclodextrin to drug ratio and copolymer to drug ratio. For each factor, natural values corresponding to the coded levels of -1, 0 and 1 were selected to cover a range of values of practical interest and based on the results of preliminary experiments conducted to assess their individual effect on the entrapment efficiency. Overall, the experimental design consisted of 13 runs. Of course, it is worth mentioning that there are 13 runs for 4 factors, which were conducted randomly to minimize the effects of uncontrolled factors. The design, layout and observed entrapment efficiency are shown in Table 1 . 2.6. In-vitro drug release The release of bosentan from nano PMs was studied using pH sensitive drug delivery system technique and using dialysis bag inside solutions with different pH. A certain amount of the drug solution trapped in nano PMs is put into the dialysis bag (cellulose membrane with mw cut off 12,400 Da) and then the dialysis bag is placed inside 50 ml of 0.1 normal hydrochloric acid with a pH of 1.2 and at a temperature of 37°C was stirred using a stirrer at 50 rpm for 2 hours and finally the solution was replaced with phosphate buffer with pH 7.4 and stirred in this solution for 24 hours. In a certain period of time, sampling of the solution was done and the amount of drug released was calculated using UV-vis, and fresh solution was added instead of the sample so that the volume of the solution does not change[ 59 – 61 ]. The cumulative release amount of drug (E n ) was calculated according to Eq. 3 . $${E}_{n}={V}_{1}\left({C}_{1}+{C}_{2}+\dots +{C}_{n-1}\right)+{V}_{0}{C}_{n}$$ 3 The cumulative rate of drug release was calculated according to Eq. 4. Cumulative release (%) = \(\frac{{E}_{n}}{{m}_{t}} \times 100\) (4) where V 1 is the volume of the drug-delivery medium, C n is the concentration of the drug in the drug-delivery medium at the n-th replacement, V 0 is the volume of the initial drug-delivery medium, and m t is the total drug amount. 3. Results and discussion 3.1. Characterization Due to the high solubility of bosentan in methanol and the use of this solvent in the drug entrapment phase, the spectrophotometric (UV-vis) spectrum of the drug was taken in methanol, which has a peak at 271 nm and can be seen in Fig. 1. In the FTIR spectrum of CD in the wave number of 3300 cm − 1 to 3400 cm − 1 , there is a broad peak that is related to the stretching of the O-H group and a strong peak at 2854 cm − 1 that is related to the symmetric/asymmetric stretching vibration of C-H, plus a peak At 1650 cm − 1 , it is related to the H-O-H bond, the change of the water in CD, and at 1153 cm − 1 , the stretching vibration related to C-O, and at 1029 cm − 1 , the C-O-C vibration of the glucose unit, and at 841 cm − 1 , the peak related to the C-O-C vibration of the cyclodextrin ring can be seen[ 62 – 65 ]. FTIR of MA–AA copolymer, the peaks at 1721 cm − 1 , 1408 cm − 1 and 2890 cm − 1 represent C = O stretching vibration, combination band of C–O stretching and O–H in plane deformation vibration and aliphatic C–H stretching vibration respectively. The peak at 1639 cm − 1 in MA–AA copolymer represents the C–C stretching vibration. These facts conclude the formation of copolymers [ 66 , 67 ]. And in the FTIR spectrum of the CD + CP composition, the peak at 1639 cm − 1 represents C–O stretching vibration, And the vibration of C-O-C is seen in 1153 cm − 1 , As well as the peak at 1408 cm − 1 represents the combination band of C–O stretching and O–H in plane deformation vibration, respectively, the peaks at 1721 cm − 1 and 1639 cm − 1 relate to the vibration C = O of the COOH group and the stretching vibration of the C-C Which represents the link between CP5 and CD. In the spectrum related to bosentan drug, the peak at 3628 cm − 1 is related to the stretching vibration of the O-H group of the free primary hydroxyl, and the weak signals in the range of 3300–3500 cm − 1 are related to the N-H stretching vibration of the secondary amine groups (-SO 2 NH-). A strong peak at 2954 cm − 1 which corresponds to aliphatic C-H stretching vibration and the peak at 1582 cm − 1 corresponding to aromatic C = C stretching vibration and asymmetric O = S stretching vibration of (-SO 2 NH-) groups at 1343 cm − 1 and 1388 cm − 1 , the peak at 1172 cm − 1 indicates the symmetric O = S stretching vibration. The O-C stretching vibration of alkyl aryl ether was observed at 1206 cm − 1 and 1251 cm − 1 . O-C stretching vibration and O-H deformation due to primary alcohol groups can be seen at 1021 cm − 1 , 1082 cm − 1 and 1113 cm − 1 [ 68 , 69 ]. In the combination of CD + CP5 + BO, the peak related to the O-H stretching vibration of free primary hydroxyl groups was reduced and a broader peak was observed at the center of 3383 cm − 1 .This indicates that the hydroxyl groups of drug molecules interact with the hydroxyl groups of polymer nanomicelles through hydrogen bonds. The peak at 2954 cm − 1 has been moved to 2932 cm − 1 . The peaks at 1343 cm − 1 and 1145 cm − 1 are due to the asymmetric and symmetric O = S stretching vibrations of (-SO 2 NH-) groups, respectively. A significant change in the O = S symmetric stretching vibration peak indicates the participation of O = S groups in hydrogen bonding with the polymer. At 1029 cm − 1 , the C-O-C vibration of the glucose unit is cyclodextrin. The index of each component is clearly visible. 3.2. Drug Entrapment Efficiency (EE) After loading the drug inside the Nano PMs and drying it inside the oven, because Nano PMs are not dissolved inside methanol but the drug is completely dissolved in methanol, 20 mL of methanol was added to it and was stirred for 20 minutes. Then centrifuged at 4500 rpm for 10 minutes, after that the supernatant was separated and UV-Vis was measured at 271 nm (Fig. 3). And by using the calibration curve, the amount of trapped drug was obtained. In the most optimal conditions, this value was 84.5%. 3.3. Calibration curve To draw the calibration curve of bosentan, concentrations from 1 to 50 mg/liter were made in methanol solvent and its absorbance was measured at 271 nm wavelength, each average point was measured three times and the equation of the line and its R 2 is was achieved to obtain the percentage of bosentan entrapment in different combinations. 3.4. Evaluation of significant variables by DSD Analysis of DSD data was carried out by a two-step procedure involving a stepwise regression followed. We could be obtained by using the stepwise selection procedure with a p-value to enter terms in the model of 0.05. The results can be seen in Table 2 . 3.5. Optimization of effective factors in drug loading Here we examine the factors together, starting with temperature and stirring time respectively. As shown in Fig. 5(a), one of the factors that strongly affects the percentage of drug entrapment is the temperature associated with the drug entrapment step. which was used to investigate the effect of temperature on drug entrapment in the temperature range of 25–55°C. According to the results, as the temperature increases up to about 43°C, the percentage of drug entrapment efficiency increases but then decreases, and this is probably due to the effect of temperature degradation on the micelle. Another parameter that has a significant effect on the percentage of drug loading is the reaction time and drug mixing with Nano PMs. This work was done for 1 to 6 hours and the percentage of drug entrapment efficiency was measured. As shown in Figu. 5(a), the loading rate increases with increasing reaction time until the maximum drug entrapment is reached after 4.25 hours of reaction, after which no increase in drug entrapment percentage is observed. Another thing that was investigated was the weight ratio of cyclodextrin to the drug, which was evaluated in ratios of 1 to 3, and the entrapment efficiency percentage was measured. As shown in Fig. 5 (b), the highest percentage of drug loading is obtained at a ratio of 2.4 to 1 cyclodextrin to drug, which is probably related to the fact that the molecular mass of cyclodextrin to drug is almost 2. The next parameter that has had a great impact is the weight ratio of the copolymer to the bosentan drug, which has been investigated in the range of 1 to 6, and as can be seen in Fig. 5 (c), with the increase in the ratio of the copolymer to the drug, the entrapment percentage increases until the ratio 4.7 that the maximum entrapment of the drug is obtained, which is most likely related to the micelle formation conditions for the bosentan drug. The pairwise interaction of the parameters can be seen in Fig. 5 (d-f). In optimal conditions, the amount of drug entrapment was 84.5%. According to the results obtained in the optimal conditions of Table 3 and the value of the standard deviation, it was determined that the design of the experiment performed with the practical test corresponds to the predicted range. 3.6. In-vitro drug release The release of bosentan drug was investigated in the first stage for 2 hours in the acidic environment of 0.1 normal hydrochloric acid solution and then in the alkaline environment of phosphate buffer inside the dialysis bag and at a temperature of 37°C. They were taken out and analyzed using a UV-Visible spectrometer to determine the release rate of bosentan. As can be seen in Fig. 6 (a), the release amount of bosentan drug in the first two hours in the acidic environment (pH 1.2) is less than 3%, and that is because the polymer part of the compound Synthesized in an acidic environment, it becomes a closed sphere that keeps the drug in itself and does not allow the release of the drug. But after these 2 hours, the release takes place in the phosphate buffer environment and pH 7.4. As it is clear in Fig. 6 (b), the release in this environment is done at a faster and controlled rate, and this is because the micelles formed in this environment are opened and the drug is released in a controlled manner, and the amount of 92.34% release takes place in phosphate buffer environment and pH 7.4. As you can see in Fig. 6 (c), further dissolution was done in phosphate buffer with pH 7.4, and after 24 hours, 95% of the drug is released, which indicates the inhibitory effect of the polymer on the formulation, which indicates The high efficiency of the synthesized compound is in the controlled transfer of bosentan drug. It is worth mentioning that for each environment, a separate calibration curve was drawn along with the release conditions, and the release percentage was obtained with the calibration curve of the same environment. 3.7. Drug release mechanism As shown in Fig. 7, the mechanism of the drug is that the micelles formed by the surface layer of which are cyclodextrin, and its internal layer is a copolymer and a drug. At pH 1.2, it is completely closed and does not allow the release of drugs. At pH 7.4, the micelle begins to fully open and the drug is released in a controlled manner. Thus the pH of the surrounding medium relative to respective p K a and p K b values of the pendant groups describe the swelling of polyacids with acidic or basic pendant groups. For the anionic matrix (with − COOH groups), if the medium pH is more than the p K a of acidic groups of polymers, there occurs ionization of acidic groups of the polymer matrix resulting in the formation of set negative charges (− COO − ) on the polymer backbone and the positive charges (H + ) are mobile in the fluid medium. Hence, there exists an electrostatic repulsion between the polymer chains leading to the swelling of the matrix and at the same time de-swelling occurs if pH is less than the p K a [ 70 , 71 ] The drug-release behaviors of nano PMs micelles with different pH values are illustrated in Fig. 7. 4. Conclusion We synthesized pH-sensitive polymer nanomicelles, it was found that it is possible to optimize the formulation of drugs with a simple and cheap method, which is better to be done in a controlled and targeted way to release the drug in order to reduce their side effects and make them more effective. A research was carried out on the transfer of bosentan drug. In the first stage, the entrapment conditions of the drug inside Nano PMs and the effective factors in entrapment were investigated by experimental design, and in the optimal conditions, the entrapment percentage of bosentan drug in Nano PMs reached 84.5%, and in the next stage, the release of the drug was investigated in laboratory conditions at pH 1.2 and 7.4. Due to the pH sensitive micelles, the lowest drug release was performed at acidic pH, which is similar to the conditions of the stomach, and more than 92% of the drug was released at pH 7.4. The orderly and purposeful face was released. Finally, we concluded that bosentan causes serious liver damage with conventional treatment, which could be reduced by developing a sustained-release formulation, which may improve patient compliance. The present research proves that Nano PMs can be a promising and potential formulation for the formulation of bosentan in providing effective and targeted treatment. Declarations Acknowledgement We acknowledge the financial support from University of Guilan (Rasht, Iran). Competing interests All authors, including, Nina Alizadeh and Hossein Ameli, affirm that they have no conflict of interest. Ethical approval This article does not involve any studies with human or animal subjects. Consent to participate Not applicable. Consent for publication Not applicable. References Chopra, H.K., Ram, C.V.S.: Recent Guidelines for Hypertension: A Clarion Call for Blood Pressure Control in India Circulation Research, Lippincott Williams and Wilkins. Circ. Res. 124 (7), 984–986 (2019). 10.1161/CIRCRESAHA.119.314789 Fuchs, F.D., Whelton, P.K.: High Blood Pressure and Cardiovascular Disease Hypertension, Lippincott Williams and Wilkins. Hypertension. 75 (2), 285–292 (2020). 10.1161/HYPERTENSIONAHA.119.14240 Tsai, H., Sung, Y.K., de Jesus, V., Perez: Recent advances in the management of pulmonary arterial hypertension. 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Controlled Release. 104 (2), 289–300 (2005) Gaucher, G.: Polymeric micelles for oral drug delivery. Eur. J. Pharm. Biopharm. 76 (2), 147–158 (2010). https://doi.org/10.1016/j.ejpb.2010.06.007 Ghezzi, M., Pescina, S.: Polymeric micelles in drug delivery: An insight of the techniques for their characterization and assessment in biorelevant conditions. J. Controlled Release. 332 , 312–336 (2021). https://doi.org/10.1016/j.jconrel.2021.02.031 Choi, J.S., Cho, N.H.: Comparison of paclitaxel solid dispersion and polymeric micelles for improved oral bioavailability and in vitro anti-cancer effects. Mater. Sci. Engineering: C. 100 , 247–259 (2019). https://doi.org/10.1016/j.msec.2019.03.002 Wang, X.: Preparation and evaluation of carboxymethyl chitosan-rhein polymeric micelles with synergistic antitumor effect for oral delivery of paclitaxel. Carbohydr. Polym. 206 , 121–131 (2019). 10.1016/j.carbpol.2018.10.096 Epub 2018 Oct 29 Kumar, R., Sirvi, A.: Polymeric micelles based on amphiphilic oleic acid modified carboxymethyl chitosan for oral drug delivery of bcs class iv compound: Intestinal permeability and pharmacokinetic evaluation. Eur. J. Pharm. Sci. 153 , 105466 (2020). https://doi.org/10.1016/j.ejps.2020.105466 Hu, W.Y., Wu, Z.M.: Smart pH-responsive polymeric micelles for programmed oral delivery of insulin. Colloids Surf. B: Biointerfaces. 183 , 110443 (2019). 10.1016/j.colsurfb.2019.110443 Nachtsheim, J.B.: A class of Three-Level Designs for Definitive Screening in the Presence of Second-Order Effects. J. Qual. Technol. 43 , 1–15 (2011) Saha, M., Saha, D.R.: QbD based development of resveratrol-loaded mucoadhesive lecithin/chitosan DOI:10.1016/j.jddst.2021.102480 nanoparticles for prolonged ocular drug delivery. J. Drug Deliv. Sci. Technol. 63 (4), 10248 (2021) Sh, R., Gabbay, R.S., Kenett: Synchronizing the release rates of salicylate and indomethacin from degradable chitosan hydrogel and its optimization by definitive screening design. Eur. J. Pharm. Sci. 125 , 102–109 (2018). Epub 2018 Sep 27 Goldman, J.M., More, H.T.: Optimization of Primary Drying in Lyophilization During Early-Phase Drug Development Using a Definitive Screening Design With Formulation and Process Factors. J. Pharm. Sci. 107 , 2592–2600 (2018). 10.1016/j.xphs.2018.06.001 Epub 2018 Jun 8 Zhang, L., Zhang, Q.: Drug-in-cyclodextrin-in-liposomes: A novel drug delivery system for flurbiprofen. Int. J. Pharm. 492 , 40–45 (2015). Epub 2015 Jul 7 Gao, W., Chan, J.M.: pH-Responsive Nanoparticles for Drug Delivery, Molecular Pharmaceutics. 7 1913–1920. (2010). https://doi.org/10.1021/mp100253e Liu, J., Huang, Y.: pH-Sensitive nano-systems for drug delivery in cancer therapy. Biotechnol. Adv. 32 , 693–710 (2014). 10.1016/j.biotechadv.2013.11.009.10 Wang, Z., Deng, X.: Mechanisms of drug release in pH-sensitive micelles for tumor targeted drug delivery system. Int. J. Pharm. 535 , 1–34 (2017). 10.1016/j.ijpharm.2017.11.003 Yallapu, M.M., Jaggi, M.: β-Cyclodextrin-curcumin self-assembly enhances curcumin delivery in prostate cancer cells. Colloids Surf. B: Biointerfaces. 79 (1), 113–125 (2010). 10.1016/j.colsurfb.2010.03.039 Sambasevam, K.P., Mohamad, S.: Synthesis and Characterization of the Inclusion Complex of β-cyclodextrin and Azomethine. Int. J. Mol. Sci. 14 , 3671–3682 (2013) Sivakumar, K., Parinamachivayam, G.: Preparation, characterization and molecular modeling studies of the beta-cyclodextrin inclusion complex with benzoguanamine and its analytical application as chemosensor for the selective sensing of Ce 4+ ,. 200 Spectrochim Acta A Mol Biomol Spectrosc. 212–225. doi: 10.1016/j.saa.2018.04.034. (2018). Epub 2018 Apr 17. Moreira, A., Bittencourt, V.: Hydrophobic nanoprecipitates of β cyclodextrin/avermectins inclusion compounds reveal insecticide activity against Aedes aegypti larvae and low toxicity against fibroblasts. J. Agric. Food Chem. 66 , 7275–7285 (2018). https://doi.org/10.1021/acs.jafc.8b01300 Senthilmurugan, B., Ghosh, B.: Maleic acid based scale inhibitors for calcium sulfate scale inhibition in high temperature application. J. Petrol. Sci. Eng. 75 , 189–195 (2010). https://doi.org/10.1016/j.petrol.2010.11.002 Rivas, B.L., Seguel, G.V.: Poly (acrylic acid-co-maleic acid)–metal complexes with copper, cobalt, and nickel Synthesis, characterization and structure of its metal chelates. Polyhedron. 18 , 2511–2518 (1999) Dangre, P.V., Sormare, B.V.: Improvement in dissolution of bosentan monohydrate by solid dispersions using spray drying technique. Open. Pharm. Sci. J. 4 , 23–31 (2017). 10.2174/1874844901704010023 Krupa, A., Majda, D.: Physicochemical properties of bosentan and selected PDE-5 inhibitors in the design of drugs for rare diseases. AAPS Pharm. Sci. Tech. 18 (4), 1318–1330 (2017). 10.1208/s12249-016-0599-7 Kulkarni, R.V., Inamdar, S.Z.: Polysaccharide-based stimuli- 7 sensitive graft copolymers for drug delivery. Edition: 1st, Chapter: 7 155–177. (2019). 10.1016/B978-0-08-102553-6.00007-6 Ameli, H., Alizadeh, N.: Targeted delivery of Capecitabine to colon cancer cells using nano polymeric micelles based on beta cyclodextrin. RSC Adv. 12 , 4681–4691 (2022). https://doi.org/10.1039/D1RA07791K Tables Table 1 Experimental design layouts and observed response (y) Run A:Temperature B:Time C:CD/B D:CP/B EE Unit °C h ratio ratio % 1 55 3.5 1 6 54.59 2 25 1 3 3.5 35.23 3 55 6 1 3.5 53.02 4 55 1 3 6 44.14 5 25 1 1 6 25.07 6 25 6 2 6 60.34 7 25 6 1 1 20.57 8 55 1 2 1 22.61 9 40 3.5 2 3.5 79.51 10 40 6 3 6 68.09 11 55 6 3 1 44.16 12 40 1 1 1 16.28 13 25 3.5 3 1 43.47 Table 2 . Estimates of the regression coefficients of the model Term Coef p-Value Intercept 78.91 - A-Temperature 3.38 0.0208 B-Time 10.29 0.0004 C-CD/B 6.56 0.002 D-CP/B 10.51 0.0003 Table 3 Results of the repeatability of the test and compliance with the design of experiments Experiment Entrapment Efficiency % Average SD Acceptable range Theory 1 82.67 82.48 2.25 80.23 – 84.72 84.5 2 84.62 3 80.14 Additional Declarations No competing interests reported. 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Alizadeh","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/klEQVRIiWNgGAWjYHACxgMJQKJBgvkAA4OBBVTQAL8eqBY2IGUgQaQWBrAWHpAyCfxKQcCcgfnBgQd/7GT7pXs+v+YpkJAHijz8wFBwD6cWywY2gwMJPMnGM+ec3WbNYyBhuLOBzViCwaAYpxaD+w+AWiSYEzfcyN1mDNTCuOEAgxlQPAG3lgPsHw4kGNQn7r+R8wykxX7DAfZvBLTwAG1JOJy4QSKH+TFQS+KGAzyEbOEpOJBw4LjxjBtpZoxzDCSSNxzmKZZIwO+wjQ9//KmW7Z+R/PjDmz82thuOt2/88OEPbi3IgA0SKcxATJwGoNoPRCocBaNgFIyCEQYAMT1UbwvomugAAAAASUVORK5CYII=","orcid":"","institution":"University of Guilan","correspondingAuthor":true,"prefix":"","firstName":"Nina","middleName":"","lastName":"Alizadeh","suffix":""}],"badges":[],"createdAt":"2024-04-06 13:44:26","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4227684/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4227684/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":54510714,"identity":"d6430d1f-802b-4c17-aa01-f9db9136b33b","added_by":"auto","created_at":"2024-04-11 15:26:56","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":24127,"visible":true,"origin":"","legend":"\u003cp\u003eUV – Vis of Bosentan in methanol\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/9bbd771aed0a5485dcb3e356.png"},{"id":54510016,"identity":"493f4089-196b-4070-9eb1-38c96a2954f6","added_by":"auto","created_at":"2024-04-11 15:18:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":95204,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectra of βCD, copolymer, CD+ Copolymer, Bosentan and CD + Copolymer + Bosentan complex.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/20e0a55d44ed62aac76d1903.png"},{"id":54510018,"identity":"536b3ba1-ee24-42b8-b7fa-64a4bc3787e5","added_by":"auto","created_at":"2024-04-11 15:18:56","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":28721,"visible":true,"origin":"","legend":"\u003cp\u003eUV spectrum of maximum drug entrapment in Nano PMs\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/057e552c44be8ce353ba37d8.png"},{"id":54510019,"identity":"0c962975-f984-4848-bbd5-7a7be8bf38fe","added_by":"auto","created_at":"2024-04-11 15:18:56","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":18673,"visible":true,"origin":"","legend":"\u003cp\u003eBosentan calibration curve in methanol solvent\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/7f55ea6ca186e13968723ba1.png"},{"id":54510020,"identity":"bb32360d-80a7-487f-93dd-57e917fa888c","added_by":"auto","created_at":"2024-04-11 15:18:56","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":180490,"visible":true,"origin":"","legend":"\u003cp\u003eshowing the simultaneous effects and 3D surface and contour plots of R (%) response for Drug Entrapment Efficiency by Nano PMs\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/e382e7a8459805fd01cfec6b.png"},{"id":54510021,"identity":"82c016e6-7e34-466c-b245-701c8f3b8eb8","added_by":"auto","created_at":"2024-04-11 15:18:56","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":28808,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eReleasing the Bosentan drug in (a) acidic environment, (b) alkaline environment and (c) pH 1.2 and pH 7.4.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/ad6a0b9f8dfc30a6b39cd8d9.png"},{"id":54510022,"identity":"8841c443-5c19-4cc3-97cb-119ce40531a1","added_by":"auto","created_at":"2024-04-11 15:18:56","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":17012,"visible":true,"origin":"","legend":"\u003cp\u003eMechanism of Bosentan drug release from the micelles\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/355baa159f0e2223137f3233.png"},{"id":55264666,"identity":"9d7063a1-69f5-440d-8d28-c6f7123880b5","added_by":"auto","created_at":"2024-04-25 01:46:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":995932,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4227684/v1/49446d4d-0650-4848-9332-ac905249b1b6.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Using nano polymeric micelles based on beta cyclodextrin for the targeted delivery of bosentan drug","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCardiovascular disease (CVD) causes almost 18\u0026nbsp;million deaths per annum. Along with strong evidence, hypertension associated with high blood pressure have a high-risk factor for CVD. Among the types of hypertension, pulmonary arterial hypertension (PAH) is perilous with 20% mortality. It results in right coronary failure and death[\u003cspan additionalcitationids=\"CR2 CR3\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePulmonary arterial hypertension (PAH) is a life threatening disease that affects about 3\u0026nbsp;million people per year. It is often fatal with a mean life about three years after diagnosis [\u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Symptoms comprise shortness of breath, chest pain, syncope, fatigue and peripheral edema which is more prevalent in females than in males [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Patients with PAH suffer from elevated levels of endothelin (ET1); a potent blood vessel constrictor, in their plasma and lung tissues [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. ET1 reduces pulmonary arterial lumen diameter, increases pulmonary vascular resistance, decreases reactivity of the vascular bed and eventually increases pulmonary arterial pressure [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Currently, endothelin receptor antagonists (ERA) are used for the treatment of PAH. Bosentan (tracleer) was the first orally active ERA to reach the clinical investigation stage [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Bosentan is a dual ERA with high affinity of both ETA and ETB receptors. Bosentan is indicated for the treatment of pulmonary artery hypertension (PAH) by blocking the action of endothelin molecule [\u003cspan additionalcitationids=\"CR14\" citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] and it can prevent or reverse the associated histological change caused by elevated levels of ET1. Bosentan and its metabolites are eliminated predominantly in the liver by the cytochrome P450 isoenzymes, CYP2C9, CYP3A4 and the terminal half-life after oral administration is 5.4 h [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. It is available in two concentrations 62.5 mg and 125 mg (for twice daily administration) and its main side effects are headache, flushing and increased liver aminotransferases. Hepatotoxicity caused by bosentan is dose-dependent in higher dosages [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe poorly soluble drugs typically limit their absorption after oral administration, ultimately limiting clinical usefulness. Bosentan is a poorly soluble oral dual endothelin receptor antagonist, usually indicated for the treatment of pulmonary arterial hypertension (PAH) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. It shows an absolute bioavailability of about 50% with plasma elimination half-life of 5.4 h [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eConsidering its low aqueous solubility, slow dissolution rate in gastrointestinal fluids, short duration of action, frequent oral administration, and dose-dependent hepatotoxicity [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], nanomicellar -based delivery system may be a better option. Nanomicellar exhibit great promise as drug delivery carriers due to their well-documented thermodynamic stability, improved capacity of solubilization, relatively low viscosity, and steric protection against protein adsorption and increased capability of cellular uptake than the free drug [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. The self-assembled nanomicelles are composed of a hydrophobic core surrounded by a hydrophilic shell (corona) of polymer, which provide effective steric protection and enable them to remain in dispersed state, prohibit recognition by mononuclear phagocyte system (MPS), ultimately stabilizing the nanomicellar structures. The prolonged circulation of micellar assists in maintaining the required therapeutic level of drug in the blood over an extended period [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. The incorporation of lipophilic drugs into the core of polymeric micellar could enhance water solubility, enhanced permeability across the physiological barriers and cause substantial improvement in oral bioavailability, and therapeutic activity with consequent reduction in dose-dependent side effects associated with the drug. As drug carriers, polysaccharidebased core-shell nanomicellar systems have demonstrated great potential to improve the delivery of hydrophobic drugs through enhanced solubility, stability, and controllable drug release properties. Various hydrophobic moieties have been conjugated to the backbone of these polysaccharides to impart them amphiphilic character. These include but not limited to cholesterol [\u003cspan additionalcitationids=\"CR25\" citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e], deoxycholic acid [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e], stearic acid [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], octane or dodecane [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e], oleoyl group [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], cetyl group [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e], and so on. Despite a significant progress, the micellar systems are still under development to search for novel biomaterials and meet the clinical need.\u003c/p\u003e \u003cp\u003eCyclodextrins are naturally available water-soluble cyclic oligosaccharides-1,4-linked-glucopyranose composed of six or more glucose units [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Cyclodextrins are well known for their ability to increase solubility, dissolution rate and bioavailability of the loaded drugs [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. They have a ring structure with a hydrophilic outer surface and lipophilic cavity which provide them with the ability to form non-covalent inclusion complexes with drug molecules of appropriate size. Inclusion complexes are very well explored to improve the water solubility and chemical stability of the drugs [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The nano polymeric micelles comprise a drug-loading core and a hydrophilic shell. Amphiphilic block copolymer forms micelles when in contact with an aqueous vehicle by self-assembly resulting in hydrophobic interactions wherein hydrophobic drugs can be encapsulated into the central core of micelles through hydrophobic interactions [\u003cspan additionalcitationids=\"CR38\" citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. The presence of this hydrophilic corona makes the micelles more stable and stealthy, helping them to avoid the reticular-endothelial system, also providing prolonged circulation and residence time in the bloodstream. These advantages, together with their small size (100 nm), make the polymeric micelles promising carriers for the administration of various insoluble and poorly soluble pharmaceutics that can be incorporated into the hydrophobic core of the micelles [\u003cspan additionalcitationids=\"CR41 CR42\" citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Another advantage of the polymeric micelles is their ability to form various types of pH-responsive drug delivery systems depending on the encapsulated drug and physiological destination. The Incorporation of pH-sensitive groups into the core-forming blocks can make the polymeric micelles sensitive to the environmental pH. In the most common strategy, ionization of the pH-sensitive groups of the inner blocks converts the micelle core from hydrophobic to hydrophilic, resulting in the demicellization of the copolymers and rapid release of the encapsulated hydrophobic drug. Polymeric micelles containing acidic groups in their inner core can be used to design smart oral drug delivery systems [\u003cspan additionalcitationids=\"CR45 CR46 CR47\" citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Such assemblies are stable at the acidic pH of the stomach because their inner blocks are in their unionized and hydrophobic form. Upon a pH increase in the intestine, the acidic groups start deprotonation which increases the electrostatic charge and hydrophilicity of the inner part, leading to the micelle dissociation and drug release. The key element for the pH sensitivity of a polymer in the presence of ionizable pendant groups which attach to the hydrophobic backbone of the polymer chain. pH-sensitive polymers are, thus, a class of polyelectrolytes with ionic functional groups that are weak acidic (\u003cem\u003ee.g.\u003c/em\u003e, carboxylic and sulfonic acids) or basic (\u003cem\u003ee.g.\u003c/em\u003e, amines, imidazole, and pyridine) groups [\u003cspan additionalcitationids=\"CR50 CR51 CR52\" citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe Copolymer of Maleic and Acrylic Acid is a copolymer of maleic anhydride and acrylic acid. It does not degrade below pH 7 thus providing film coats that are resistant to gastric media but soluble in intestinal fluid. A combination of Nano PMs and β-CD can improve the drug loading capacity and therapeutic efficacy of poorly water soluble drugs. In the analysis of problems affected by several factors with possible interactions, statistical screening methods are often adopted to select the parameters that actually affect the response variable and to eliminate the irrelevant ones. This is particularly useful at the beginning of an investigation when little or no information is available for the system of interest. Recently, a new class of three-level designs, the so-called definitive screening designs (DSD), has been proposed by Jones and Nachtsheim [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. DSD allows for assessment of active effects, two-factor interactions and pure-quadratic effects in the presence of effector sparsity. This allows a dramatic reduction in the number of experiments, thus enabling a significant saving in time and cost of materials. The use of these screening methods is increasing in recent years [\u003cspan additionalcitationids=\"CR56\" citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. Thus, the objective of this present investigation was to synthesize pH-sensitive nano polymeric micelles as controlled release carriers for bosentan.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003e2.1. Materials\u003c/h2\u003e\n \u003cp\u003e\u0026beta;- Cyclodextrin (purity, \u0026gt;\u0026thinsp;98%) was purchased from Sigma, The drug Bosentan monohydrate (purity, \u0026gt;\u0026thinsp;99%) was taken from Abmole Company, Acrylic/maleic copolymer (purity, \u0026gt;\u0026thinsp;92%) With a molar mass of 70,000 g/mol And brand CP 5 powder was purchased from BASF Germany, dialysis membrane (MWCO 12 kDa) was purchased from Sigma, and all other reagents and chemicals were of analytical grade and purchased from sigma.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003e2.2. Characterization\u003c/h2\u003e\n \u003cp\u003eThe UV-Vis absorption measurements were taken at room temperature using UV 1800 spectrophotometer (SHIMADZU, Japan). The UV-Vis of the samples were recorded in the frequency range of 200\u0026thinsp;~\u0026thinsp;700 nm. The FTIR spectra of the Nanocomposites-bosentan nanoparticles and free Bosentan were obtained with a (protege 460) FTIR Spectrometer (Nicolet, USA). For FTIR spectroscopic investigations, 10 mg of the sample were mixed with 100 mg KBr and pressed into a pellet. The measurements were carried out in the mid-infrared range of 4000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e to 400 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The resolution was set to 4 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; 100 scans were recorded, averaged for each spectrum.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003e2.3. Preparation of Bosentan loaded Nano polymeric micelles\u003c/h2\u003e\n \u003cp\u003eThe inclusion complex of bosentan, cyclodextrin and copolymer was prepared using the evaporation method with solvents. Equivalent amounts of pure drug, cyclodextrin and copolymer were dissolved in 15 ml of methanol and 85 ml of distilled water, respectively. This solution was placed in an ultrasonic bath for 15 minutes. The solution was transferred to a conical flask and stirred using a magnetic stirrer for 4 hours at a suitable temperature at 150 rpm. The resulting solution was evaporated at a temperature of 45\u0026deg;C and the dried complex was passed through sieve No. 44[\u003cspan class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003e2.4. Determining the amount of drug and entrapment efficiency of bosentan\u003c/h2\u003e\n \u003cp\u003eTo measure the amount of drug trapped in nano polymeric micelles (Nano polymeric micelles), we take a certain amount of it and dissolve it in methanol, because nano PMs are not dissolved in methanol and only the drug bosentan that is not trapped is dissolved in methanol. Then it is centrifuged and the concentration of the free drug in the supernatant is measured using a UV-Vis device at a wavelength of 271 nm, and as a result, the amount of trapped drug is obtained by Eqs.\u0026nbsp;1 and 2.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e \u003csub\u003e\u0026nbsp;\u003cstrong\u003ee\u003c/strong\u003e\u0026nbsp;\u003c/sub\u003e \u003cstrong\u003e= C\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003ei\u003c/strong\u003e\u003c/sub\u003e \u003cstrong\u003e- C\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003es\u003c/strong\u003e\u003c/sub\u003e (1)\u003c/p\u003e\n \u003cp\u003eEE (entrapment efficiency) \u003cstrong\u003e=\u003c/strong\u003e \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\frac{\\mathbf{C}\\mathbf{e}}{\\mathbf{C}\\mathbf{i}}\\times 100\\)\u003c/span\u003e\u003c/span\u003e (2)\u003c/p\u003e\n \u003cp\u003eWhere, C\u003csub\u003ee\u003c/sub\u003e is the Amount of drug trapped (encapsulated); \u003cstrong\u003eC\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003ei\u003c/strong\u003e\u003c/sub\u003e is the Initial drug value; \u003cstrong\u003eC\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003es\u003c/strong\u003e\u003c/sub\u003e is the drug value in the supernatant; EE is the drug entrapment efficiency.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003e2.5. Experimental design\u003c/h2\u003e\n \u003cp\u003eThe DSD proposed by Jones and Nachtsheim [\u003cspan class=\"CitationRef\"\u003e54\u003c/span\u003e] was adopted to investigate the effects of 4 continuous factors (k\u0026thinsp;=\u0026thinsp;4) for Bos-loaded Nano polymeric micelles that were identified, in preliminary runs, as potentially important for the drug loading process. They were temperature, time, cyclodextrin to drug ratio and copolymer to drug ratio. For each factor, natural values corresponding to the coded levels of -1, 0 and 1 were selected to cover a range of values of practical interest and based on the results of preliminary experiments conducted to assess their individual effect on the entrapment efficiency. Overall, the experimental design consisted of 13 runs. Of course, it is worth mentioning that there are 13 runs for 4 factors, which were conducted randomly to minimize the effects of uncontrolled factors. The design, layout and observed entrapment efficiency are shown in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003e2.6. In-vitro drug release\u003c/h2\u003e\n \u003cp\u003eThe release of bosentan from nano PMs was studied using pH sensitive drug delivery system technique and using dialysis bag inside solutions with different pH. A certain amount of the drug solution trapped in nano PMs is put into the dialysis bag (cellulose membrane with mw cut off 12,400 Da) and then the dialysis bag is placed inside 50 ml of 0.1 normal hydrochloric acid with a pH of 1.2 and at a temperature of 37\u0026deg;C was stirred using a stirrer at 50 rpm for 2 hours and finally the solution was replaced with phosphate buffer with pH 7.4 and stirred in this solution for 24 hours. In a certain period of time, sampling of the solution was done and the amount of drug released was calculated using UV-vis, and fresh solution was added instead of the sample so that the volume of the solution does not change[\u003cspan class=\"CitationRef\"\u003e59\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e61\u003c/span\u003e]. The cumulative release amount of drug (E\u003csub\u003en\u003c/sub\u003e) was calculated according to Eq.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e\n \u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\n \u003cdiv class=\"mathdisplay\" id=\"FileID_Equ1\" name=\"EquationSource\"\u003e$${E}_{n}={V}_{1}\\left({C}_{1}+{C}_{2}+\\dots +{C}_{n-1}\\right)+{V}_{0}{C}_{n}$$\u003c/div\u003e\n \u003cdiv class=\"EquationNumber\"\u003e3\u003c/div\u003e\n \u003c/div\u003e\n \u003cp\u003eThe cumulative rate of drug release was calculated according to Eq.\u0026nbsp;4.\u003c/p\u003e\n \u003cp\u003eCumulative release (%) = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\frac{{E}_{n}}{{m}_{t}} \\times 100\\)\u003c/span\u003e\u003c/span\u003e (4) where V\u003csub\u003e1\u003c/sub\u003e is the volume of the drug-delivery medium, C\u003csub\u003en\u003c/sub\u003e is the concentration of the drug in the drug-delivery medium at the n-th replacement, V\u003csub\u003e0\u003c/sub\u003e is the volume of the initial drug-delivery medium, and m\u003csub\u003et\u003c/sub\u003e is the total drug amount.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"3. Results and discussion","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1. Characterization\u003c/h2\u003e\n \u003cp\u003eDue to the high solubility of bosentan in methanol and the use of this solvent in the drug entrapment phase, the spectrophotometric (UV-vis) spectrum of the drug was taken in methanol, which has a peak at 271 nm and can be seen in Fig. 1.\u003c/p\u003e\n \u003cp\u003eIn the FTIR spectrum of CD in the wave number of 3300 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e to 3400 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, there is a broad peak that is related to the stretching of the O-H group and a strong peak at 2854 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e that is related to the symmetric/asymmetric stretching vibration of C-H, plus a peak At 1650 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, it is related to the H-O-H bond, the change of the water in CD, and at 1153 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, the stretching vibration related to C-O, and at 1029 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, the C-O-C vibration of the glucose unit, and at 841 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, the peak related to the C-O-C vibration of the cyclodextrin ring can be seen[\u003cspan class=\"CitationRef\"\u003e62\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e65\u003c/span\u003e]. FTIR of MA\u0026ndash;AA copolymer, the peaks at 1721 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 1408 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 2890 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e represent C\u0026thinsp;=\u0026thinsp;O stretching vibration, combination band of C\u0026ndash;O stretching and O\u0026ndash;H in plane deformation vibration and aliphatic C\u0026ndash;H stretching vibration respectively. The peak at 1639 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in MA\u0026ndash;AA copolymer represents the C\u0026ndash;C stretching vibration. These facts conclude the formation of copolymers [\u003cspan class=\"CitationRef\"\u003e66\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e67\u003c/span\u003e]. And in the FTIR spectrum of the CD\u0026thinsp;+\u0026thinsp;CP composition, the peak at 1639 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e represents C\u0026ndash;O stretching vibration, And the vibration of C-O-C is seen in 1153 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, As well as the peak at 1408 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e represents the combination band of C\u0026ndash;O stretching and O\u0026ndash;H in plane deformation vibration, respectively, the peaks at 1721 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1639 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e relate to the vibration C\u0026thinsp;=\u0026thinsp;O of the COOH group and the stretching vibration of the C-C Which represents the link between CP5 and CD. In the spectrum related to bosentan drug, the peak at 3628 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e is related to the stretching vibration of the O-H group of the free primary hydroxyl, and the weak signals in the range of 3300\u0026ndash;3500 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e are related to the N-H stretching vibration of the secondary amine groups (-SO\u003csub\u003e2\u003c/sub\u003eNH-). A strong peak at 2954 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e which corresponds to aliphatic C-H stretching vibration and the peak at 1582 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e corresponding to aromatic C\u0026thinsp;=\u0026thinsp;C stretching vibration and asymmetric O\u0026thinsp;=\u0026thinsp;S stretching vibration of (-SO\u003csub\u003e2\u003c/sub\u003eNH-) groups at 1343 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1388 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, the peak at 1172 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicates the symmetric O\u0026thinsp;=\u0026thinsp;S stretching vibration. The O-C stretching vibration of alkyl aryl ether was observed at 1206 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1251 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. O-C stretching vibration and O-H deformation due to primary alcohol groups can be seen at 1021 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 1082 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1113 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [\u003cspan class=\"CitationRef\"\u003e68\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e69\u003c/span\u003e]. In the combination of CD\u0026thinsp;+\u0026thinsp;CP5\u0026thinsp;+\u0026thinsp;BO, the peak related to the O-H stretching vibration of free primary hydroxyl groups was reduced and a broader peak was observed at the center of 3383 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.This indicates that the hydroxyl groups of drug molecules interact with the hydroxyl groups of polymer nanomicelles through hydrogen bonds. The peak at 2954 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e has been moved to 2932 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The peaks at 1343 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1145 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e are due to the asymmetric and symmetric O\u0026thinsp;=\u0026thinsp;S stretching vibrations of (-SO\u003csub\u003e2\u003c/sub\u003eNH-) groups, respectively. A significant change in the O\u0026thinsp;=\u0026thinsp;S symmetric stretching vibration peak indicates the participation of O\u0026thinsp;=\u0026thinsp;S groups in hydrogen bonding with the polymer. At 1029 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, the C-O-C vibration of the glucose unit is cyclodextrin. The index of each component is clearly visible.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2. Drug Entrapment Efficiency (EE)\u003c/h2\u003e\n \u003cp\u003eAfter loading the drug inside the Nano PMs and drying it inside the oven, because Nano PMs are not dissolved inside methanol but the drug is completely dissolved in methanol, 20 mL of methanol was added to it and was stirred for 20 minutes. Then centrifuged at 4500 rpm for 10 minutes, after that the supernatant was separated and UV-Vis was measured at 271 nm (Fig. 3). And by using the calibration curve, the amount of trapped drug was obtained. In the most optimal conditions, this value was 84.5%.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003e3.3. Calibration curve\u003c/h2\u003e\n \u003cp\u003eTo draw the calibration curve of bosentan, concentrations from 1 to 50 mg/liter were made in methanol solvent and its absorbance was measured at 271 nm wavelength, each average point was measured three times and the equation of the line and its R\u003csup\u003e2\u003c/sup\u003e is was achieved to obtain the percentage of bosentan entrapment in different combinations.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003e3.4. Evaluation of significant variables by DSD\u003c/h2\u003e\n \u003cp\u003eAnalysis of DSD data was carried out by a two-step procedure involving a stepwise regression followed. We could be obtained by using the stepwise selection procedure with a p-value to enter terms in the model of 0.05. The results can be seen in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003e3.5. Optimization of effective factors in drug loading\u003c/h2\u003e\n \u003cp\u003eHere we examine the factors together, starting with temperature and stirring time respectively. As shown in Fig. 5(a), one of the factors that strongly affects the percentage of drug entrapment is the temperature associated with the drug entrapment step. which was used to investigate the effect of temperature on drug entrapment in the temperature range of 25\u0026ndash;55\u0026deg;C. According to the results, as the temperature increases up to about 43\u0026deg;C, the percentage of drug entrapment efficiency increases but then decreases, and this is probably due to the effect of temperature degradation on the micelle. Another parameter that has a significant effect on the percentage of drug loading is the reaction time and drug mixing with Nano PMs. This work was done for 1 to 6 hours and the percentage of drug entrapment efficiency was measured. As shown in Figu. 5(a), the loading rate increases with increasing reaction time until the maximum drug entrapment is reached after 4.25 hours of reaction, after which no increase in drug entrapment percentage is observed. Another thing that was investigated was the weight ratio of cyclodextrin to the drug, which was evaluated in ratios of 1 to 3, and the entrapment efficiency percentage was measured. As shown in Fig. 5 (b), the highest percentage of drug loading is obtained at a ratio of 2.4 to 1 cyclodextrin to drug, which is probably related to the fact that the molecular mass of cyclodextrin to drug is almost 2. The next parameter that has had a great impact is the weight ratio of the copolymer to the bosentan drug, which has been investigated in the range of 1 to 6, and as can be seen in Fig. 5 (c), with the increase in the ratio of the copolymer to the drug, the entrapment percentage increases until the ratio 4.7 that the maximum entrapment of the drug is obtained, which is most likely related to the micelle formation conditions for the bosentan drug. The pairwise interaction of the parameters can be seen in Fig. 5 (d-f). In optimal conditions, the amount of drug entrapment was 84.5%. According to the results obtained in the optimal conditions of Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e and the value of the standard deviation, it was determined that the design of the experiment performed with the practical test corresponds to the predicted range.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003e3.6. In-vitro drug release\u003c/h2\u003e\n \u003cp\u003eThe release of bosentan drug was investigated in the first stage for 2 hours in the acidic environment of 0.1 normal hydrochloric acid solution and then in the alkaline environment of phosphate buffer inside the dialysis bag and at a temperature of 37\u0026deg;C. They were taken out and analyzed using a UV-Visible spectrometer to determine the release rate of bosentan. As can be seen in Fig. 6 (a), the release amount of bosentan drug in the first two hours in the acidic environment (pH 1.2) is less than 3%, and that is because the polymer part of the compound Synthesized in an acidic environment, it becomes a closed sphere that keeps the drug in itself and does not allow the release of the drug.\u003c/p\u003e\n \u003cp\u003eBut after these 2 hours, the release takes place in the phosphate buffer environment and pH 7.4. As it is clear in Fig.\u0026nbsp;6 (b), the release in this environment is done at a faster and controlled rate, and this is because the micelles formed in this environment are opened and the drug is released in a controlled manner, and the amount of 92.34% release takes place in phosphate buffer environment and pH 7.4. As you can see in Fig.\u0026nbsp;6 (c), further dissolution was done in phosphate buffer with pH 7.4, and after 24 hours, 95% of the drug is released, which indicates the inhibitory effect of the polymer on the formulation, which indicates The high efficiency of the synthesized compound is in the controlled transfer of bosentan drug. It is worth mentioning that for each environment, a separate calibration curve was drawn along with the release conditions, and the release percentage was obtained with the calibration curve of the same environment.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003e3.7. Drug release mechanism\u003c/h2\u003e\n \u003cp\u003eAs shown in Fig.\u0026nbsp;7, the mechanism of the drug is that the micelles formed by the surface layer of which are cyclodextrin, and its internal layer is a copolymer and a drug. At pH 1.2, it is completely closed and does not allow the release of drugs. At pH 7.4, the micelle begins to fully open and the drug is released in a controlled manner. Thus the pH of the surrounding medium relative to respective p\u003cem\u003eK\u003c/em\u003e\u003csub\u003ea\u003c/sub\u003e and p\u003cem\u003eK\u003c/em\u003e\u003csub\u003eb\u003c/sub\u003e values of the pendant groups describe the swelling of polyacids with acidic or basic pendant groups. For the anionic matrix (with \u0026minus;\u0026thinsp;COOH groups), if the medium pH is more than the p\u003cem\u003eK\u003c/em\u003e\u003csub\u003ea\u003c/sub\u003e of acidic groups of polymers, there occurs ionization of acidic groups of the polymer matrix resulting in the formation of set negative charges (\u0026minus;\u0026thinsp;COO\u003csup\u003e\u0026minus;\u003c/sup\u003e) on the polymer backbone and the positive charges (H\u003csup\u003e+\u003c/sup\u003e) are mobile in the fluid medium. Hence, there exists an electrostatic repulsion between the polymer chains leading to the swelling of the matrix and at the same time de-swelling occurs if pH is less than the p\u003cem\u003eK\u003c/em\u003e\u003csub\u003ea\u003c/sub\u003e [\u003cspan class=\"CitationRef\"\u003e70\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e71\u003c/span\u003e] The drug-release behaviors of nano PMs micelles with different pH values are illustrated in Fig. 7.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003eWe synthesized pH-sensitive polymer nanomicelles, it was found that it is possible to optimize the formulation of drugs with a simple and cheap method, which is better to be done in a controlled and targeted way to release the drug in order to reduce their side effects and make them more effective. A research was carried out on the transfer of bosentan drug. In the first stage, the entrapment conditions of the drug inside Nano PMs and the effective factors in entrapment were investigated by experimental design, and in the optimal conditions, the entrapment percentage of bosentan drug in Nano PMs reached 84.5%, and in the next stage, the release of the drug was investigated in laboratory conditions at pH 1.2 and 7.4. Due to the pH sensitive micelles, the lowest drug release was performed at acidic pH, which is similar to the conditions of the stomach, and more than 92% of the drug was released at pH 7.4. The orderly and purposeful face was released. Finally, we concluded that bosentan causes serious liver damage with conventional treatment, which could be reduced by developing a sustained-release formulation, which may improve patient compliance. The present research proves that Nano PMs can be a promising and potential formulation for the formulation of bosentan in providing effective and targeted treatment.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe acknowledge the financial support from University of Guilan (Rasht, Iran).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors, including,\u0026nbsp;Nina Alizadeh and Hossein Ameli,\u0026nbsp;affirm that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis article does not involve any studies with human or animal subjects.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eChopra, H.K., Ram, C.V.S.: Recent Guidelines for Hypertension: A Clarion Call for Blood Pressure Control in India Circulation Research, Lippincott Williams and Wilkins. 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RSC Adv. \u003cb\u003e12\u003c/b\u003e, 4681\u0026ndash;4691 (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/D1RA07791K\u003c/span\u003e\u003cspan address=\"10.1039/D1RA07791K\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cdiv align=\"Left\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"425\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"6\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Experimental design layouts and observed response (y)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003eRun\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003eA:Temperature\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003eB:Time\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003eC:CD/B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003eD:CP/B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003eEE\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003eUnit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026deg;C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003eh\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003eratio\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003eratio\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e54.59\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e35.23\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e53.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e44.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e25.07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e60.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e20.57\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e22.61\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e79.51\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e68.09\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e44.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e16.28\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.705882352941178%\" valign=\"top\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.294117647058824%\" valign=\"top\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16%\" valign=\"top\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.529411764705882%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.176470588235293%\" valign=\"top\"\u003e\n \u003cp\u003e43.47\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cdiv align=\"Left\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"309\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"4\" valign=\"bottom\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e Estimates of the regression coefficients of the model\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.65695792880259%\" valign=\"bottom\"\u003e\n \u003cp\u003eTerm\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.359223300970875%\" colspan=\"2\" valign=\"bottom\"\u003e\n \u003cp\u003eCoef\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"44.983818770226534%\" valign=\"bottom\"\u003e\n \u003cp\u003ep-Value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.65695792880259%\" valign=\"top\"\u003e\n \u003cp\u003eIntercept\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.359223300970875%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e78.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"44.983818770226534%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.65695792880259%\"\u003e\n \u003cp\u003eA-Temperature\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.359223300970875%\" colspan=\"2\"\u003e\n \u003cp\u003e3.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"44.983818770226534%\"\u003e\n \u003cp\u003e0.0208\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.16129032258065%\" colspan=\"2\"\u003e\n \u003cp\u003eB-Time\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e10.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"44.83870967741935%\"\u003e\n \u003cp\u003e0.0004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.16129032258065%\" colspan=\"2\"\u003e\n \u003cp\u003eC-CD/B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e6.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"44.83870967741935%\"\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.16129032258065%\" colspan=\"2\"\u003e\n \u003cp\u003eD-CP/B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20%\"\u003e\n \u003cp\u003e10.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"44.83870967741935%\"\u003e\n \u003cp\u003e0.0003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\n \u0026nbsp;\n\u003c/p\u003e\n\u003cdiv align=\"Left\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"505\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"6\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 3\u003c/strong\u003e Results of the repeatability of the test and compliance with the design of experiments\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.648221343873518%\"\u003e\n \u003cp\u003eExperiment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.019762845849803%\"\u003e\n \u003cp\u003eEntrapment Efficiency %\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.079051383399209%\"\u003e\n \u003cp\u003eAverage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.521739130434782%\"\u003e\n \u003cp\u003eSD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.272727272727273%\"\u003e\n \u003cp\u003eAcceptable range\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.458498023715414%\"\u003e\n \u003cp\u003eTheory\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.648221343873518%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.019762845849803%\"\u003e\n \u003cp\u003e82.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.079051383399209%\" rowspan=\"3\"\u003e\n \u003cp\u003e82.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.521739130434782%\" rowspan=\"3\"\u003e\n \u003cp\u003e2.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.272727272727273%\" rowspan=\"3\"\u003e\n \u003cp\u003e80.23 \u0026ndash; 84.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.458498023715414%\" rowspan=\"3\"\u003e\n \u003cp\u003e84.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"45.714285714285715%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"54.285714285714285%\"\u003e\n \u003cp\u003e84.62\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"45.714285714285715%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"54.285714285714285%\"\u003e\n \u003cp\u003e80.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Bosentan, Nano polymeric micelles, Cyclodextrin, Definitive Screening Design","lastPublishedDoi":"10.21203/rs.3.rs-4227684/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4227684/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe drugs formulation have been considered based on the micelles of polymeric nanoparticles in the past decades. It has shown that these Nano polymeric micelles (Nano PMs) help to increase accessibility to homeland, decreasing side effects and getting long of drug dissemination. The purpose of this study is optimization of the bosentan drug formula using polymer nanomicelles and cyclodextrins. The pH-responsive copolymer was prepared and the variables of loading time, loading temperature, the ratio of CP5 copolymer to drug and the ratio of cyclodextrin to drug and the effect of these variables on the percentage of drug loading were investigated. In this work, first, a Definitive Screening Design was used to investigate the effect of variables on drug loading percentage. This method provides the possibility of significantly reducing the number of tests required to check systems that are characterized by a large number of variables. In the optimized formula for the drug, the loading temperature was 43 °C and the time was 4.25 hours, as well as the ratio of CP5 copolymer to the drug was 4.7 and the ratio of cyclodextrin to the drug was 2.4 to reach the maximum efficiency of 84.5% drug entrapment. The release was carried out in laboratory conditions at pH 1.2 and 7.4 and at a temperature of 37 °C for 24 hours, 95% of the drug was released, which showed that the release of the drug was completely targeted and controlled. Therefore, according to the obtained results, we can hope that polymer nanomicelles with cyclodextrin are potential and efficient carriers for targeted delivery with sustained release of drugs.\u003c/p\u003e","manuscriptTitle":"Using nano polymeric micelles based on beta cyclodextrin for the targeted delivery of bosentan drug","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-11 15:18:51","doi":"10.21203/rs.3.rs-4227684/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b4d4df72-a70f-4fee-a438-f4d1b64835dc","owner":[],"postedDate":"April 11th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-04-22T07:15:06+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-11 15:18:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4227684","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4227684","identity":"rs-4227684","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}