Development and Optimization of Thermoreversible in-situ Nasal Gel Loaded With Zavegepant for Treatment of Migraine

Development and Optimization of Thermoreversible in-situ Nasal Gel Loaded With Zavegepant for Treatment of Migraine | 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 Development and Optimization of Thermoreversible in-situ Nasal Gel Loaded With Zavegepant for Treatment of Migraine Anil R. Pawar, Vishal Balme, Ramdas B. Pandhare, Vinayak K. Deshmukh, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7281614/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 Objectives: To develop and optimize a thermoreversible in-situ nasal gel of Zavegepant for effective and rapid treatment of acute migraine, enhancing brain targeting and bioavailability while overcoming limitations of oral formulations. Methods: A 3² full factorial design was employed to evaluate the effects of Pluronic F-127 (X₁) and xanthan gum (X₂) on gelation temperature (Y₁) and mucoadhesive strength (Y₂). Nine formulations (VF1–VF9) were developed and evaluated for physicochemical properties, gelation behavior, mucoadhesion, in-vitro drug release, ex vivo permeation, and in vivo anti-migraine efficacy using a nitroglycerin-induced migraine model in rats. Results: Optimized batch VF2 containing 20% Pluronic F-127 and 0.2% xanthan gum showed a gelation temperature of 34.94 °C and mucoadhesive strength of 5812.2 dyne/cm² with minimal prediction error (<5%). VF2 exhibited sustained ex vivo drug release (83.67% at 8 hours) and steady-state flux of 522.94 μg/cm²/h. In vivo studies demonstrated significant improvement in locomotor activity, photophobia, and mechanical allodynia, with biochemical normalization of CGRP (41.16 pg/mg), MDA, NO, GSH, and SOD levels, comparable to sumatriptan. Stability over 3 months confirmed formulation robustness. Conclusion: The optimized thermosensitive nasal gel (VF2) of Zavegepant presents a promising, non-invasive strategy for acute migraine therapy with sustained drug release, enhanced mucosal retention, and effective CNS delivery. Its clinical potential lies in offering fast, localized treatment with fewer systemic side effects and improved patient compliance. Zavegepant in-situ nasal gel migraine factorial design CGRP antagonist intranasal delivery thermoreversible gel pharmacodynamics Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 INTRODUCTION Migraine is a significant health issue in the world, with a world prevalence level of migraine disability of about 14% of all the world population, with higher risk in a group of population where women are more vulnerable at the rate of 18% and men 9% [ 1 ]. As a neurological disorder, it has a massive economic cost to the United States, of around $ 27 billion a year in direct care costs and productivity loss [ 2 ]. The existing methods of treatment are not fully effective, as the relief provided by the current drugs fails to cope with the necessity in approximately 40% of patients. Other limitations included in these factors are delayed onset of action, large first-pass metabolism, and GI disorders which are relatively problematic during nausea experienced with migraines. Treatment gap is especially high in the middle and low-income countries where proportion of migraine patients who obtain proper treatment is less than 7–14% [ 3 ]. Migraine is also given by periodical attacks of moderate to severe headache, which is always combined with the presence of nausea and vomiting, photosensitivity (sensitivity to light), and phonophobia (sensitivity to sound) [ 4 ]. There are numerous triggers of migraine although it is not clearly understood; these triggers occur in the form of genetic predisposition, environmental factors, hormonal shifts and dysfunction of the brain [ 5 ]. The pathophysiology of migraines has a great dependence on the activation of the trigeminovascular system that subsequently releases neuropeptides such as calcitonin gene-related peptide (CGRP). This leads to vasodilation, neurogenic inflammation and intensification of the nerve endings pain response. Migraines with aura have also been attributed to cortical spreading depression, a neuronal depolarization wave through the cerebral cortex [ 6 ]. Zavegepant is a new calcitonin gene-related peptide (CGRP) receptor antagonist in the acute treatment of migraine structure of zavegepant is depicted in Fig. 1 [ 7 ]. It is the first and only CGRP antagonist as a nasal spray formulation that can be used to obtain immediate relief to symptoms of migraine [ 8 ]. Zavegepant has an advantage of quick absorption via the nose since oral CGRP inhibitors take time to react against nausea and vomiting in migraine patients [ 9 ]. The mechanism of action of the zavegepant drug is inhibition of CGRP receptors and prevention of vasodilation and neuroinflammation of migraine pathophysiology. Recent clinical trials indicate that, zavegepant can initially relieve pain and symptom-free in two hours and has few safety or tolerability concerns [ 10 ]. One of the potentially successful systems to deliver the drug treating migraine is thermosensitive in situ gels applied to the nasal route [ 11 ]. The formulations are in liquid form at the room temperature and change to gel when they are in contact with nasal mucosa at a body temperature. This peculiarity leads to better retention of drugs to the location of drug absorption and better bioavailability and therapeutic effect [ 12 ]. The most widely used thermosensitive polymer is poloxamer 407 (Pluronic F-127), which is a nonionic triblock copolymer synthesised using polyethylene oxide (PEO) and polypropylene oxide (PPO) [ 13 ]. Poloxamer 407 is temperature-responsive to form gels and therefore this properly gels at physiological temperature leaving the drug to stay long in the nasal cavity [ 14 ]. Nasal route provides the multiple advantages such as rapid absorption, ease of administration, avoids first pass and etc. Moreover, the drugs can enter through the central nervous system via the olfactory and trigeminal nerve routes. Thermosensitive in situ nasal gels are specifically appealing to the treatment of acute migraine attacks because of a need of a sweeping speed of action [ 15 ]. This is the major goal of the study; to invent and perfect a thermoreversible in situ nasal gel system that incorporates zavegepant to treat migraines. The pursuit of this study is to improve the bioavailability and residence time of the drug in the nasal cavity by using a unique system of gel that can exist in liquid state at room temperature but become gel at the temperature existing within the nasal cavity. This new method of drug delivery will enhance adherence in the patient, results in the fast onset of action, and continuous slow release of a drug combined with the avoidance of first-pass metabolism and the direct delivery of zavegepant via nose to the brain. MATERIALS AND METHODS Materials Zavegepant (≥98% purity) was procured from Pfizer Inc. (New York, USA). Pluronic F-127 (pharmaceutical grade) and xanthan gum (food grade, particle size <150 μm) were sourced from Research Lab Fine Chem Industries (Mumbai, India). Polyethylene glycol 400 (PEG 400, pharmaceutical grade) and Benzalkonium chloride (0.01% w/v, preservative grade) were purchased from Merck Life Science Pvt. Ltd. (Mumbai, India). Nitroglycerin solution (5 mg/mL injection) was obtained from Sun Pharmaceutical Industries Ltd. (Mumbai, India). Sumatriptan succinate (>99% purity), HPLC-grade acetonitrile, methanol, and water were purchased from Sigma-Aldrich (Mumbai, India). Potassium bromide (KBr) (spectroscopic grade) for FTIR analysis was supplied by Himedia Laboratories (Mumbai, India). All other chemicals and reagents used were of analytical grade and used without further purification. METHODS Calibration Curve of Zavegepant UV-visible spectrophotometer (Shimadzu UV-1800, Shimadzu India, Mumbai) was used to prepare a calibration curve of zavegepant. Zavegepant (10 mg) was reconstituted in 100 mL methanol to produce a stock solution of 100 1259038 g/mL. Standard solutions of 5, 10, 15, 20, 25 and 30 0g/ml were prepared by proper dilution with methanol. The solutions were measured as Absorbance at 239 nm λ max against methanol blank. The plot of the absorbance with respect to the concentration was used to form the calibration curve that was used to get the linear regression equation. Correlation coefficient (R 2 ), slope and y- intercept were used to validate the linearity. Percent standard deviation of response as well as slope was used in calculating LOD and LOQ. Each measurement was done in triplicate (n=3) [16,17]. Solubility Study The saturation shake flask method was used to determine the solubility of zavegepant in five solvents such as methanol, ethanol, distilled water, DMSO, and phosphate buffer at pH 6.8. Surplus quantities of the drug (10 mg) were placed in 10 mL of each of solvents in closed vials. A 24 hour orbital shaking at 100 rpm was carried at 25 ± 2°C in orbital incubator (REMI CIS-24 BL, India). Once in equilibrium, the samples were centrifuged at 5000 rpm and after 15 minutes filtration was performed using 0.45 mm membrane filter. To mix, the filtrate was diluted properly with methanol and analyzed through the UV-visible spectrophotometer at 239 nm. Each of the experiments was repeated three times (n=3) and the means of solubility values were reported as mean±SD [18,19]. Differential Scanning Calorimetry (DSC) The thermal properties of zavegepant and possible drug-excipient interaction were studied using differential scanning calorimetry. Analysis was done through a DSC-60 Plus (Shimadzu Corporation, Kyoto, Japan). Pure zavegepant and physical mixture were weighed accurately (5 mg) and placed in standard aluminum pans and heated between 30-300 o C with heating rate of 10 o C/min with nitrogen atmosphere (50 mL/min flow rate). A reference was an empty sealed aluminum pan. The alteration of endothermic peak characteristics was tested to determine the interaction of drug-excipient and alteration of crystallinity structure. All the measurements were done three-fold (n=3) [20,21]. Fourier Transform Infrared (FTIR) Spectroscopy Analysis of chemical interactions between zavegepant and formulation components Chemical interactions between zavegepant and formulation components were examined by FTIR spectroscopy using FTIR spectrophotometer (IRAffinity-1S, Shimadzu Corporation, Kyoto, Japan) with DTGS detector. Pure zavegepant, single polymers, and physical mixtures were prepared by separately mixing 2 mg of each sample with 100 mg dry KBr and pressed into their transparent pellets using a hydraulic pressure in a vacuum. Spectra in the range of 4000-400 cm -1 were obtained with a resolution of 4 cm -1 . This measurement was done in triplicates (n=3). Frequencies of peak shift, vanishing, or broadening patterns were beforehand revealed in order to identify drug-excipient interactions [22,23]. Optimization of In-situ Nasal Gel A 3² full factorial design was employed to systematically investigate the influence of two critical formulation variables: Pluronic F-127 (X₁), a thermosensitive gelling agent, and xanthan gum (X₂), a mucoadhesive polymer, on the performance of the in-situ nasal gel. Each factor was evaluated at three levels, resulting in nine experimental runs. The independent variables (X₁ and X₂) were assessed for their effects on two key dependent responses: gelation temperature (Y₁) and mucoadhesive strength (Y₂). The design layout, including both actual and coded values, is presented in Tables 1 and 2. The relationship between formulation variables and responses was fitted to a second-order polynomial equation: Y = b₀ + b₁X₁ + b₂X₂ + b₁₂X₁X₂ + b₁₁X₁² + b₂₂X₂² where Y denotes the predicted response, X₁ and X₂ are the coded levels of independent variables, and coefficients represent the magnitude and direction of influence. Analysis of variance (ANOVA) was performed to determine statistical significance (p<0.05). The optimal formulation was identified using desirability function with constraints set as gelation temperature (32-36°C) and maximized mucoadhesive strength [24,25]. Table 1: Design Matrix of 3² Factorial Study Highlighting Independent Factors, Coded Levels, Dependent Measures, and Optimization Targets Factors (Independent Variables) Levels Low (-1) Medium (0) High (+1) A: Pluronic F-127 (%) 18 20 22 B: Xanthan gum (%) 0.1 0.2 0.3 Dependent Variables Goal Y₁: Gelation Temperature (°C) 30-34°C Y₂: Mucoadhesive Strength (N/cm²) Maximize Table 2: Ingredient Profile of Zavegepant-Incorporated In-Situ Nasal Gel Formulations(VF1-VF9) Ingredients VF1 VF2 VF3 VF4 VF5 VF6 VF7 VF8 VF9 Zavegepant (mg) 10 10 10 10 10 10 10 10 10 Pluronic F-127 (%w/v) 18 20 22 18 20 22 18 20 22 Xanthan gum (%w/v) 0.1 0.1 0.1 0.2 0.2 0.2 0.3 0.3 0.3 Benzalkonium chloride (%) 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 PEG 400 (%) 1 1 1 1 1 1 1 1 1 Distilled water q.s. to 10 mL q.s. to 10 mL q.s. to 10 mL q.s. to 10 mL q.s. to 10 mL q.s. to 10 mL q.s. to 10 mL q.s. to 10 mL q.s. to 10 mL Preparation of Zavegepant-loaded In-situ Nasal Gel The cold method was used in the preparation of the in-situ nasal gel. Pre-cooled phosphate buffer (pH 6.4 and 4 C) was gradually added to Pluronic F-127 and this was stirred continuously at 400 rpm (Remi Equipment Pvt. Ltd., Mumbai, India) The dispersion was re-frigerated-after full hydration-at 4±1 o C, over a 24 hours period. Xanthan gum was dispersed individually into phosphate buffer and eventually put in Pluronic solution continuously till the mixture was stirred. The dissolved solution of Zavegepant (10 mg) was mixed into the PEG 400 and polymer solution. Benzalkonium chloride (0.01%) was added as preservative. To complete the final volume, phosphate buffer was added up to 10 mL and the samples were stored at 4±1 o C within 24 hours and then evaluated. Each formulation was prepared in three replicates (n=3) and the pH was set between 6.4±0.2 [26]. Evaluation of Thermosensitive In-situ Nasal Gel pH Determination In order to make sure that the pH in situ nasal gel needed to be suitable to apply in the nasal system, a digital pH meter (Eutech pH 700, Singapore) was used. 1 g of the formulation was dispersed in 10 mL of distilled water and the electrode inserted to measure the pH when it reached its normal level. pH 4.0, pH 7.0 and pH 9.2 buffer solutions of standard concentration were used to calibrate the instrument. Measurements were made three times (n=3) at 25±2 o C [27]. Gelation Temperature The tube inversion method of determining gelation temperature was used. A water bath temperature was pre-set to 20 o C by placing a 5 mL formulation in a test tube. The temperature was raised at a rate of 1 o C/minute with calibrated thermometers being used to track this process. The test tube was turned upside down at 90 o angle at every temperature increment and observation of the formation of gel was made. The point at which the formulation ceased to flow has been noted as the gelation temperature. Each measurement was done three times (n=3) [28]. Gelation Time Gelation time was recorded in a slightly altered method to that of Miller and Donovan method. An amount of 2 mL of gel formulation was placed in 1.0 cm (diameter) test tube and sealed with Parafilm, and vertical setting in a circulating water bath at 37 o C temperature. Following a 10-minutes equilibration period, the tube was placed in a horizontal position to note the formation of gels. The sol-to-gel transition was found at the moment when the formulation ceased to flow. The experimental operations were repeated thrice (n=3) [29]. Viscosity Measurement Measures of viscosity of the in-situ nasal gel pre and post gelation were grabbed using a Brookfield viscometer (LVDV-E, Brookfield Engineering Labs) where spindle No. 64 (Technosys Instruments, Thane, India) was used. The 10 mL formulation was used to measure at 25±0.5 o C and 34 ± 0.5°C. During the test, the spindle speed was kept at 50 rpm. Measurement was carried out after equilibrating in a 34 ± 1°C water bath at 20 minutes after gelation. The values of viscosity were measured in centipoise (cP). All the measurements were carried out three times (n=3) [30]. Spreadability Engineering variables were calculated by slip and drag method to determine the spreadability. Clean glass slides (7.5 x 2.5 cm) were used to put the properly weighed gel in between them. A 100 g weight was pressed on the upper slide to make it evenly distributed by 5 minutes. A wooden block was used to support the lower slide. The automatic separation of the upper slide following the removal of the weights (t) was recorded. Based on the following formula, spreadability (S) was calculated: S = (M × L)/T where M is applied weight (g), L is slide length (cm), and T is time (sec). All measurements were performed in triplicate (n=3) [31]. Drug Content Uniformity Drug content was analyzed as: gel (equivalent to 1 mg zavegepant) was dissolved in 10 mL of methanol with vortexing at 5 minutes. The solution was diluted and then filtered using 0.45 μm membrane filter. UV-visible spectrophotometer was used to examine absorbance in 239 nm, with blank consisting of methanol. The concentration of drugs would be determined based on the calibration curve already obtained. Each of the tests was repeated thrice (n=3) [32]. Mucoadhesive Strength The modified balance (Figure 2) technique was adopted to test the mucoadhesive strength on new excised goat nasal mucus. Mucosal tissues of 1 cm 2 in size were taken and were washed using distilled water and phosphate buffer (pH 6.4) to flake out the connective tissue. The mucosa including the mucosal side were attached to glass with cyanoacrylate adhesive. The support was put in phosphate buffer solution on the right hand pan of a physical balance. The weight placed at the left pan was 5 g in order to get the equilibrium. Gel was used as 1g and it was used to completely cover the mucosa and this was done in 2min. Gradually water was added to the left pan till detachment of mucosa occurred in the gel. Mucoadhesive strength was measured in terms of the weight of detachment, and converted in number of dyne/cm 2 . The experiments were done thrice (n=3) [33]. Ex Vivo Permeation Study Franz diffusion cells were used in permeation studies where goat nasal mucosa was used. The mucosal tissue after cleaning was evenly sliced and placed between donor and receptor chambers and the mucosal surface exposed toward the donor. The receptor compartment was filled with 20 mL of phosphate buffer (pH 6.4) that was kept at 37±0.5 o C and stirred continuously at 100 rpm. The donor chamber was filled with gel formulation 1 g. Aliquots (1 mL) of the receptor medium were removed at specific times (0, 1, 2, 3, 4, 5, 6, 7 and 8 hours) and replaced with fresh buffer to ensure sink conditions. The samples were filtered by 0.45 2003 um membrane filter and UV-visible spectrophotometer was used to analyze with reference to 239 nm. The Jss and the Kp were determined based on the linear part of the permeation curve. Each experiment was done with triplicates (n=3) [34,35]. Animals and Ethical Clearance The male Wistar rats were maintained in polypropylene cages under controlled of temperature (25±2 o C), relative humidity (60±5 percent)) in standard laboratory conditions 12:12 light/dark. Food was standard pellet diet and ad libitum water. The study was done after one week of acclimatization. All animal studies were carried out on the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) and approved by the Institutional Animal Ethics Committee (IAEC) (Approval No. MESCOP/IAEC/2023-24/01). Proper steps were taken that would ensure the minimum suffering of animals and the least number of animals are utilized [36]. Nitroglycerin (NTG)-Induced Migraine Model Nitroglycerin (NTG)-induced migraine model in rats was used, because the developed zavegepant-loaded in-situ nasal gel had been tested on a validated rat model of migraine before. It was found that NTG solution (5 mg/kg, subcutaneously, injected in the dorsal region) caused migraine-like symptoms. In post-NTG animals, the trends of commonly observed migraine behaviour were simulated such as a motor dysfunction, photophobia, phonophobia and allodynia on mechanical stimulus. The results of these responses were quantified by common accepted conductions of behaviors: a locomoter activity was assessed through the open field test, photo phobia was experienced through the light dark box test as well as mechanical allodynia through the von frey filament approach. Symptoms akin to migraine usually appeared in 1 hour after injection of NTG and thereafter interventions were taken to treat it in accordance with the experiment design [37,38]. Experimental Design The animals were randomized into five groups containing 6 animals each (n = 6) and evaluated to assess the anti-migraine activity of: Group I (Normal Control): intravenously administered saline; Group II (Migraine Control): intravenously given nitroglycerin (NTG, 5 mg/kg, s.c.) with no treatment; Group III (Standard Treatment): NTG (5 mg/kg, s.c.) given with an intranasal dose of sumatriptan (6 mg/kg); Group IV given NTG (5 mg/kg, In the case of intranasal administration, 20 μL of respective formulation was administered into each nostril with a micropipette equipped with a low-density polyethylene tube in the animal supine position under mild (2-3% of isoflurane) anesthesia line. The behavioral tests were made at 0 hour (baseline), 1 hour (post-NTG, pre-treatment), 2, 3 and 4 hours after injection of NTG. The experimentation ended with the death of animals by exposing them to CO 2 inhalation, and the brain tissues were harvested in order to be checked on biochemical and pathological level [39,40]. RESULTS AND DISCUSSION Calibration Curve of Zavegepant The calibration plot of zavegepant was a straight line with correlation coefficient (R 2 ) of 0.9991 in the range of 5-30 μg/mL. The quantitative analysis showed strong enlightenment of good linearity and the ability of the equation Y = 0.0182x + 0.0091 to support quantitative analysis (Figure 3). It was established that limit of detection (LOD) and limit of quantification (LOQ) was 0.15 μg/mL and 0.45 μg/mL, respectively. Solubility Study Solubility of Zavegepant in various solvents was largely different. The maximum solubility was seen in DMSO (112.8±3.45 mg/mL), methanol (45.6±1.02 mg/mL), ethanol (12.5±0.43 mg/mL), phosphate buffer pH 6.8 (4.3±0.21 mg/mL), and water (0.06±0.01 mg/mL). Zavegepant was freely soluble in DMSO, soluble in methanol, sparingly soluble in ethanol, slightly soluble in the phosphate buffer and practically insoluble in the water as per the Indian Pharmacopoeia classification (Table 3). Table 3: Solubility study of zavegepant in different solvents Sr. No. Solvent Solubility (mg/mL) Classification 1 Water 0.06 ± 0.01 Practically insoluble 2 Ethanol 12.5 ± 0.43 Sparingly soluble 3 Methanol 45.6 ± 1.02 Soluble 4 Phosphate Buffer pH 6.8 4.3 ± 0.21 Slightly soluble 5 DMSO 112.8 ± 3.45 Freely soluble Values expressed as mean ± SD (n=3) Differential Scanning Calorimetry (DSC) Pure zavegepant showed a sharp endothermic peak at 264.07 o C, which showed that it is crystalline. Here the physical mixture had two endothermic peaks located at 262.13 o C and 275.5 o C which represented zavegepant and excipients respectively (Figure 4). The fact that no major peak shifts, broadening or disappearance occurred was indicative that there was no significant physicochemical interaction between the drug and excipients which implied close compatibility with formulation development. FTIR Spectroscopy Pure zavegepant revealed the characteristic FTIR spectra of 3281 cm -1 (N-H stretching), 2934.3 and 2850.5 cm -1 (C-H stretching), 1680.0 cm -1 (C=O stretching) as well as 1608.5 cm -1 (aromatic C=C stretching). Peaks observed on physical mixture were comparable at 3310.8 cm -1 (N-H), 2932.0 -1 and 2857.7cm -1 (C-H), 1689.9 -1 and 1724.0 cm -1 (C=O) and 1608.0 cm -1 (C=C) (Figure 5, Table 4) with no drastic changes. This ascertained the non-existence of a chemical interaction between zavegepant and excipients. Table 4: FTIR Interpretation of Pure Zavegepant and Physical Mixture Functional Group Standard Wavenumber (cm⁻¹) Observed in Pure Drug (cm⁻¹) Observed in Physical Mixture (cm⁻¹) N-H stretching 3300-3400 3281 3310.8 C-H stretching 2850-2950 2934.3, 2850.5 2932.0, 2857.7 C=O stretching 1650-1750 1680.0 1689.9, 1724.0 Aromatic C=C stretch 1580-1620 1608.5 1608.0 C-N stretching 1200-1350 1245.3, 1280.4 1240.6, 1281.0 C-O stretching 1000-1300 1132.7, 1024.9 1131.7, 1027.6 Physicochemical Evaluation of Thermoresponsive In-Situ Nasal Gel All the nine formulations (VF1-VF9) had clear appearance, homogeneous, and good consistency, which indicated successful drug incorporation and polymer dispersion. The physical Physical Characterization The uniform appearance and a transparent homogeneous texture indicate the successful placement of drugs and dispersion of polymer in all the nine formulations (VF1-VF9) as well as their excellent consistency. This compliances with the formulations suitability in nasal delivery as revealed by physical stability and reproducibility between any two different batches (Table 5). Table 5: Physical characterization of thermosensitive in-situ nasal gel formulations Formulation Appearance Homogeneity Consistency Phase Separation VF1 Transparent Homogeneous Excellent No VF2 Transparent Homogeneous Excellent No VF3 Transparent Homogeneous Excellent No VF4 Transparent Homogeneous Excellent No VF5 Transparent Homogeneous Excellent No VF6 Transparent Homogeneous Excellent No VF7 Transparent Homogeneous Excellent No VF8 Transparent Homogeneous Excellent No VF9 Transparent Homogeneous Excellent No All physicochemical characteristics of formulations showed an appropriate profile of nasal delivery. pH values of the formulations were between 5.1 and 6.4, confirming their suitability as nasal delivery. The degree of spreadability was 7.6 ±0.76 to 11.6±0.78 cm which is good. The viscosity of all gels before the gelation point ranged between 170.25±1.63 and 284.86±1.77 cP as established in the charts of the viscosity obtained, and after the gelation point increased significantly to 11638.1±132.2 and 12342.8±102.4 to substantiate their effective thermosensitive behavior. Content uniformity of drugs was high in all formulations (94.42 to 9877%) and shows good reproducibility of manufacturing, as shown in Table 6. Table 6: Physicochemical characteristics of zavegepant-loaded in-situ gel formulations Formulation pH Spreadability (cm) Viscosity before gelation (cP) Viscosity after gelation (cP) Drug content (%) VF1 5.5 ± 0.06 8.2 ± 0.43 170.25 ± 1.63 11782.5 ± 128.6 94.42 ± 1.23 VF2 5.8 ± 0.08 7.6 ± 0.76 187.91 ± 0.98 11968.4 ± 115.4 97.32 ± 0.89 VF3 6.2 ± 0.03 8.8 ± 0.13 220.45 ± 1.75 12092.7 ± 107.3 97.67 ± 0.76 VF4 5.3 ± 0.06 9.1 ± 0.53 218.22 ± 2.89 11638.1 ± 132.2 96.56 ± 1.04 VF5 5.1 ± 0.02 11.6 ± 0.78 192.32 ± 1.31 11864.3 ± 121.7 94.86 ± 1.34 VF6 6.1 ± 0.07 8.3 ± 0.57 220.15 ± 2.78 12125.6 ± 110.8 97.45 ± 0.92 VF7 5.4 ± 0.08 10.2 ± 0.62 284.86 ± 1.77 12342.8 ± 102.4 98.74 ± 0.65 VF8 5.5 ± 0.03 9.4 ± 0.89 246.12 ± 0.42 12210.5 ± 108.6 98.77 ± 0.58 VF9 6.4 ± 0.05 8.7 ± 0.44 268.51 ± 0.18 12184.9 ± 113.2 96.52 ± 1.12 Values expressed as mean ± SD (n=3) Gelation properties exhibited concentration-dependent behavior whereby the temperature of gelation ranged between 25.22±0.85 o C and 41.02±0.28 o C and the time of gelation was between 19.5±0.5 and 35.3±0.6 seconds. Mucoadhesive strength was 4230.9±0.42 to 6895.7±0.23 dyne/cm 2 which proves sufficient retentive power to be used in the mucosa (Table 7). Table 7: Gelation temperature, gelation time, and mucoadhesive strength of thermosensitive in-situ nasal gel formulations Formulation Gelation Temperature (°C) Gelation Time (sec) Mucoadhesive Strength (dyne/cm²) VF1 39.45 ± 0.54 32.4 ± 0.7 5192.2 ± 0.65 VF2 34.94 ± 0.13 26.2 ± 0.4 5812.4 ± 0.90 VF3 26.52 ± 0.64 22.5 ± 0.2 4230.9 ± 0.42 VF4 41.02 ± 0.28 35.3 ± 0.6 5696.8 ± 0.74 VF5 36.23 ± 0.87 24.1 ± 0.2 6690.7 ± 0.32 VF6 28.04 ± 1.45 21.0 ± 0.8 5219.5 ± 0.18 VF7 39.12 ± 0.72 30.2 ± 0.6 5791.5 ± 0.94 VF8 35.75 ± 0.39 22.8 ± 0.2 6895.7 ± 0.23 VF9 25.22 ± 0.85 19.5 ± 0.5 5797.9 ± 0.56 Values expressed as mean ± SD (n=3) Statistical Optimization using 3² Factorial Design The model for gelling temperature with a quadratic function was statistically significant with a model p value of 0.0012; thus, it appeared to fit the experimental data well on the whole. The model was tested and their correlation with the model is summarized in Table 8 with Adjusted R² of 0.9863 and predicted R² of 0.9460 and this meant the model has a good capability of prediction. The concentration of PF127 (Factor A) proved to be most significant among the model terms (p = 0.0002, F = 546.59) while quadratic terms A² (p = 0.0162, F = 24.07) and B² (p = 0.0476, F = 10.55) were also significant. Nevertheless, the results of Table IX show that the linear term of xanthan gum (B) and the interaction term (AB) were not statistically significant (p > 0.5). The results suggest that gelling temperature is mainly with PF127 due to its nonlinearity. Then the final regression equation of the gelling temperature (Y₁) can be coded variables. Y₁ = 36.7044 − 6.635A − 0.1367B − 0.2425AB − 2.4117A² − 1.5967B² Figure 7 shows that gelling temperature decreased significantly with increasing concentration of PF127 and xanthan gum exhibited minimal linear effect. Following the curve along the PF127 axis, a steep decline was observed, shaping a bowl shaped curvature, supporting that the quadratic effects matter in the gelation behaviors. Effect of Formulation Variables on Mucoadhesive Strength (Y₂) Both the quadratic model for mucoadhesive strength had a highly significant p value for the model (p = 0.0002), an adjusted R² of 0.9887 and predicted R² of 0.9960 (Table 8). Both linear contributors were shown to be significant by PF 127 (F = 133.19; p = 0.0014) and xanthan gum (F = 685.70; p = 0.0001) based on ANOVA data in Table 9. Interaction (AB: F = 91.21, p = 0.0024) and both quadratic terms (A²: F = 1021.21, p < 0.0001; B²: F = 48.27, p = 0.0061) were highly significant as this indicated synergistic and curvature driven effects on mucoadhesion. Thus, the coded form of the regression equation for mucoadhesive strength (Y₂) is: Y₂ = 6632.2 − 238.7A + 541.6B + 241.925AB − 1144.8A² − 248.9B² Figure 7 illustrates that mucoadhesive strength increases with xanthan gum concentration while PF127 variability declined nonspecifically with PF127 concentration. Response surface had curved topology and contour plot depicted elliptical bands corresponding to optimal interaction of the two polymers for effective mucosal adhesion. Table 8: Model Fit Summary for Responses of Thermosensitive In-Situ Nasal Gel Formulation Response Model Type Sequential p-value Adjusted R² Predicted R² Suggested Model Gelling Temperature Linear 0.0003 0.8440 0.9131 2FI 0.8094 0.8971 0.6147 Quadratic 0.0225 0.9863 0.9460 Suggested Cubic 0.5911 0.9857 0.6735 Aliased Mucoadhesive Strength Linear 0.2022 0.2174 -0.2718 2FI 0.5431 0.1345 -1.3116 Quadratic 0.0001 0.9960 0.9887 Suggested Cubic 1.0000 0.9879 0.7242 Aliased Table 9: ANOVA for Quadratic Models of Thermosensitive In-Situ Nasal Gel Formulation Source Sum of Squares df Mean Square F-value p-value Significance Gelling Temperature Model 281.22 5 56.24 116.39 0.0012 Significant A – PF127 264.14 1 264.14 546.59 0.0002 Significant B – Xanthan Gum 0.11 1 0.11 0.23 0.6631 Not Significant AB 0.24 1 0.24 0.49 0.5356 Not Significant A² 11.63 1 11.63 24.07 0.0162 Significant B² 5.10 1 5.10 10.55 0.0476 Significant Residual 1.45 3 0.48 Total 282.67 8 Mucoadhesive Strength Model 5080996.82 5 1016199.36 395.92 0.0002 Significant A – PF127 341866.14 1 341866.14 133.19 0.0014 Significant B – Xanthan Gum 1759983.36 1 1759983.36 685.70 0.0001 Significant AB 234110.82 1 234110.82 91.21 0.0024 Significant A² 2621134.08 1 2621134.08 1021.21 <0.0001 Significant B² 123902.42 1 123902.42 48.27 0.0061 Significant Residual 7700.08 3 2566.69 Total 5088696.90 8 Validation of Statistical Model The results of the predicted value and experimental value of maximized formulation (VF2) were compared and served as the validation factor of the statistical model. Ref. the gelation temperature was expected as 34.12°C as compared to the experimental value of 34.94°C (percent error of 2.35°C). The key parameter to measure mucoadhesive strength; the predicted value was 6060.7 dyne/cm2 as compared to experimental value of 5812.2 dyne/cm 2 (relative error 4.10%). The superior correspondence of less than 5% errors proves the high predictabilities and reliabilities of quadratic equations (Table 10). Table 10: Validation of optimized zavegepant-loaded mucoadhesive in-situ nasal gel formulation Factor/Response Composition (%w/v) Predicted Value Experimental Value Relative Error (%) Formulation Variables Pluronic F-127 20.000 20.000 20.000 - Xanthan gum 0.200 0.200 0.200 - Critical Quality Attributes Gelation Temperature (°C) - 34.12 34.94 2.35 Mucoadhesive Strength (dyne/cm²) - 6060.7 5812.2 4.10 Desirability - 0.785 - - Ex Vivo Drug Permeation Study Ex vivo transfers revealed that the three zavegepant formulation formulations achieved controlled and sustained drug delivery over a period of 8 hours. VF3 also had the maximum cumulative drug release (91.28%), VF7 was next and then VF5. The optimized formulation VF2 had effective drug permeation (83.67 % at 8 hours) exhibiting appropriate release pattern in nasal drug delivery (Table 11, Figure 8). Table 11: In vitro drug release profile of zavegepant in-situ nasal gel formulations Time (hr) VF1 VF2 VF3 VF4 VF5 VF6 VF7 VF8 VF9 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1 12.45 10.86 13.78 11.92 12.84 11.45 13.24 12.93 11.83 2 24.68 22.94 26.93 23.85 25.73 22.86 25.92 24.83 23.74 3 35.92 33.67 38.56 34.76 36.82 33.94 37.83 35.92 34.85 4 46.75 43.92 49.82 45.83 47.93 44.82 48.92 46.83 45.92 5 56.84 54.76 60.94 55.92 58.84 54.93 59.83 57.92 56.83 6 68.93 65.83 72.85 67.84 70.93 65.84 71.92 69.83 67.92 7 79.26 75.92 84.73 77.93 81.74 75.93 82.84 80.74 78.83 8 87.54 83.67 91.28 85.82 89.63 84.72 90.73 88.63 86.74 Steady-state Flux and Permeability Coefficient Analysis To assess the features of drug transports by such parameters as permeation across the nasal mucosa, the calculation of the permeation parameters was performed (Table 12). VF3 exhibited the best steady-state flux (570.50 μg/cm 2 /h), and permeability coefficient (0.114 cm /h), which is indicative of a better permeation potential. The optimised formulation VF2 had steady-state flux of 522.94 μg/cm 2 /h and permeability coefficient of 0.105 cm/h which showed that the formulation met appropriate characteristics of a controlled nasal drug delivery. Table 12: Steady-state flux (Jss) and permeability coefficient (Kp) of zavegepant-loaded thermosensitive in-situ nasal gel formulations Formulation Code Steady-state Flux (μg/cm²/h) Permeability Coefficient (cm/h) VF1 547.13 ± 12.4 0.109 ± 0.003 VF2 522.94 ± 15.2 0.105 ± 0.004 VF3 570.50 ± 18.6 0.114 ± 0.005 VF4 536.38 ± 14.8 0.107 ± 0.003 VF5 560.19 ± 16.2 0.112 ± 0.004 VF6 529.50 ± 13.6 0.106 ± 0.003 VF7 567.06 ± 17.4 0.113 ± 0.004 VF8 553.94 ± 15.8 0.111 ± 0.004 VF9 542.13 ± 14.2 0.108 ± 0.003 Values expressed as mean ± SD (n=3) In Vivo Animal Studies There were notable changes in locomotor activity, photophobia, mechanical allodynia, and biochemical parameters as per nitroglycerin inspired migraine model, among the treatment groups .Induction of migraine and treatment is shown in figure10. The results are that the normal control group sustained stable locomotor activity during the experiment with line crossings of 92.8±5.9 and 96.2±6.1 at the beginning and end of the experiment, respectively whereas the migraine control group has shown significant decrease following the administration of NTG that reduces to 32.7±4.0 at 4 hours. Both locomotor scores after treatment with sumatriptan and zavegepant solution as well as the optimized in-situ nasal gel (VF2) increased substantially with an area under the curves of 82.4±7.1 and 70.2±6.6 and 80.1±7.4, respectively (Table 13). Equally, time on the light side in the photophobia test was abridged drastically in migraine control category (66.4±9.2 to 55.6±8.5 seconds) yet rats administered with sumatriptan had amplified inclination to the light that sustained up to 160.3±15.6 seconds. A significant improvement in the zavegepant solution and VF2 group was also seen, with a decrease to the levels of 130.6±14.8 and 153.1±16.5 seconds, respectively (Table 14) at the 4-hour period or time point. Mechanical allodynia showed that in normal animals the withdrawal thresholds were high (~22 g), but in the rat group subjected to migraine, withdrawal thresholds significantly decreased to 6.1±1.0 g. The zavegepant solution, sumatriptan, and VF2 groups demonstrated signs of progressive normalization of the pain thresholds with a post-operative benefit occurring at 4 hours with VF2 resulting in the threshold restoration of 18.1±2.2 g (Table 15). Biochemical test revealed that the migraine control group depicted high levels of nitric oxide (13.24±1.52 μmol/mg), MDA (7.16±0.79 nmol/mg), and CGRP (87.42±7.46 pg/mg) and reduced antioxidant markers, GSH (17.85±2.12 μmol/mg) and SOD (13.18±1.62 U/ Treatment with VF2 normalized these values a great deal and decreased the nitric oxide to 5.63 +/- 0.72, the MDA to 2.84±0.39 as well as the CGRP to 41.16±3.67 and raised again the GSH to 29.34±3.12 and SOD to 20.51±2.19 (Table 16). These results substantiated the greater neuroprotective and headache prophylactic effects of the enhanced zavegepant-filled in-situ nasal gel (VF2) whose therapeutic performance equaled that of sumatriptan and was tremendously enhanced when compared to bare zavegepant formulation (solution).all parameters are depicted in figure 9. Table 13: Effect of Different Formulations on Locomotor Activity in Nitroglycerin-induced Migraine Model Treatment Groups 0 h (Baseline) 1 h (Post-NTG) 2 h 3 h 4 h Group I: Normal Control 96.2 ± 6.1 93.7 ± 5.8 95.4 ± 6.2 94.1 ± 6.5 92.8 ± 5.9 Group II: Migraine Control 95.6 ± 5.7 40.8 ± 5.1* 36.5 ± 4.4* 34.1 ± 4.2* 32.7 ± 4.0* Group III: Standard Treatment (Sumatriptan) 94.9 ± 5.9 41.2 ± 5.0* 60.7 ± 6.0*# 76.3 ± 6.7*# 82.4 ± 7.1*# Group IV: Zavegepant Solution 96.0 ± 6.2 39.7 ± 5.3* 53.9 ± 5.6*# 64.5 ± 6.0*# 70.2 ± 6.6*# Group V: Optimized Formulation (VF2) 97.1 ± 6.4 41.5 ± 5.4* 57.6 ± 6.2*# 71.8 ± 7.0*# 80.1 ± 7.4*# *p < 0.05 compared to normal control group; #p < 0.05 compared to migraine control group Table 14: Effect of Different Formulations on Photophobia in Nitroglycerin-induced Migraine Model Treatment Groups 0 h (Baseline) 1 h (Post-NTG) 2 h 3 h 4 h Group I: Normal Control 187.4 ± 14.2 185.6 ± 13.7 186.3 ± 14.0 183.9 ± 13.4 184.5 ± 13.9 Group II: Migraine Control 185.3 ± 13.9 66.4 ± 9.2* 60.2 ± 8.7* 58.7 ± 8.3* 55.6 ± 8.5* Group III: Standard Treatment (Sumatriptan) 188.2 ± 13.6 67.1 ± 9.5* 107.2 ± 12.8*# 139.4 ± 14.9*# 160.3 ± 15.6*# Group IV: Zavegepant Solution 186.1 ± 14.3 65.9 ± 8.6* 95.7 ± 11.3*# 118.2 ± 13.6*# 130.6 ± 14.8*# Group V: Optimized Formulation (VF2) 185.8 ± 13.5 64.7 ± 9.1* 102.8 ± 12.0*# 132.4 ± 15.2*# 153.1 ± 16.5*# *p < 0.05 compared to normal control group; #p < 0.05 compared to migraine control group Table 15: Effect of Different Formulations on Mechanical Allodynia in Nitroglycerin-induced Migraine Model Treatment Groups 0 h (Baseline) 1 h (Post-NTG) 2 h 3 h 4 h Group I: Normal Control 22.1 ± 2.2 21.6 ± 2.0 22.0 ± 2.1 21.8 ± 2.3 22.3 ± 2.0 Group II: Migraine Control 22.5 ± 2.1 7.9 ± 1.3* 7.2 ± 1.2* 6.6 ± 1.1* 6.1 ± 1.0* Group III: Standard Treatment (Sumatriptan) 22.3 ± 2.0 8.2 ± 1.4* 13.7 ± 1.9*# 17.3 ± 2.0*# 19.0 ± 2.3*# Group IV: Zavegepant Solution 22.6 ± 2.2 7.8 ± 1.2* 11.6 ± 1.8*# 14.2 ± 2.0*# 15.7 ± 2.1*# Group V: Optimized Formulation (VF2) 22.4 ± 2.1 7.6 ± 1.3* 12.5 ± 1.7*# 15.9 ± 1.9*# 18.1 ± 2.2*# *p < 0.05 compared to normal control group; #p < 0.05 compared to migraine control group Table 16: Effect of Different Formulations on Biochemical Parameters in Brain Tissue in Nitroglycerin-induced Migraine Model Treatment Groups Nitric Oxide (μmol/mg protein) MDA (nmol/mg protein) GSH (μmol/mg protein) SOD (U/mg protein) CGRP (pg/mg protein) Group I: Normal Control 3.18 ± 0.34 1.79 ± 0.25 37.92 ± 3.58 25.07 ± 2.41 25.88 ± 2.96 Group II: Migraine Control 13.24 ± 1.52* 7.16 ± 0.79* 17.85 ± 2.12* 13.18 ± 1.62* 87.42 ± 7.46* Group III: Standard Treatment (Sumatriptan) 5.12 ± 0.66*# 2.63 ± 0.36*# 31.65 ± 3.38*# 21.94 ± 2.28*# 38.92 ± 3.49*# Group IV: Zavegepant Solution 7.56 ± 0.89*# 4.12 ± 0.49*# 27.02 ± 2.87*# 17.48 ± 2.03*# 52.87 ± 5.31*# Group V: Optimized Formulation (VF2) 5.63 ± 0.72*# 2.84 ± 0.39*# 29.34 ± 3.12*# 20.51 ± 2.19*# 41.16 ± 3.67*# *Results are expressed as Mean ± SD (n = 6); *p < 0.05 compared to normal control; #p < 0.05 compared to migraine control Stability Study Physical and chemical stability of the optimized formulation was demonstrated to be excellent as revealed in the accelerated stability study over the three months. The preparation had the quality of transparent appearance during the entire period of study. There was small deviation shown in the gelation temperature that varied between 34.94±0.13 0 and 32.8±0.7 and this did not experience very high rates of variation that could make it too high to apply on the nose. Drug content was good with only 4.01% decrease after three months (97.32 to 93.35%). All other parameters such as viscosity, mucoadhesive strength, and ex vivo permeation had minor changes, which were within the limits and hence affirmed the formulation robustness and potential shelf-life (Table 17). Table 17: Three-month stability study results of optimized zavegepant-loaded mucoadhesive in-situ nasal gel formulation (VF2) Parameter Initial 1 Month 2 Months 3 Months Physical appearance Clear, transparent No change No change Slightly darker pH 5.8 ± 0.08 5.7 ± 0.07 5.6 ± 0.06 5.5 ± 0.07 Gelation temperature (°C) 34.94 ± 0.13 34.2 ± 0.5 33.4 ± 0.6 32.8 ± 0.7 Drug content (%) 97.32 ± 0.89 96.8 ± 0.98 95.2 ± 1.05 93.35 ± 1.18 Viscosity before gelation (cP) 187.91 ± 0.98 185.6 ± 1.2 182.3 ± 1.4 178.5 ± 1.6 Viscosity after gelation (cP) 11968.4 ± 115.4 11756.2 ± 162.4 11542.8 ± 168.6 11328.6 ± 172.8 Mucoadhesive strength (dyne/cm²) 5812.4 ± 0.90 5689.3 ± 0.72 5542.7 ± 0.81 5398.2 ± 0.75 Ex vivo drug permeation at 8 hr (%) 83.67 ± 0.88 82.42 ± 0.73 80.16 ± 0.81 78.94 ± 0.94 Microbial testing No growth No growth No growth No growth Values expressed as mean ± SD (n=3) DISCUSSION To create an effective, non-invasive and patient friendly alternative in the management of acute migraine, a thermoreversible in-situ nasal gel loaded with zavegepant was developed. Zavegepant is a calcitonin gene-related peptide (CGRP) receptor antagonist that has good potential as a treatment and management tool in migraine treatment, since they act precisely and more strategically than other headaches with distinct effects, highly powerful, and fast acting [ 41 ]. But it has a low oral bioavailability with large first-pass metabolism and hence its use as a drug. Another effective alternative to this problem is the nasal route which has direct access to systemic circulation and the brain, and bypasses hepatic metabolism [ 42 ], through the olfactory and trigeminal systems. The present research was able to exploit the strategy of quality by design (QbD) to develop and optimize a formulation of zavegepant-loaded thermoreversible in-situ gel to be delivered intranasally. Optimization was conducted on a 3 factorial design since the concentrations of Poloxamer 407 and Poloxamer 188 were regarded as important factors affecting the gelation temperature and viscosity of the gel [ 43 ]. Poloxamer 407 is a temperature sensitive polymer that gels at nasal cavity temperature but remains in liquid state at room temperature so is easily administerable. The gelling temperature and formulation consistency is regulated by poloxamer 188. Optimized formulation VF2 had an optimal gelation temperature (~ 31.5 o C) which would be favorable in nasal administration, an adequate viscosity of 245 cP, which would result in retention and low mucociliary clearance after any administration. The physical stability and equal distribution of zavegepant was ascertained by the clarity, the pH and the drug content of the formulation being within acceptable limits [ 44 ]. According to the in vitro drug release studies, optimized VF2 formulation had a profile of a sustained release as opposed to that of the pure drug solution. VF2 obeyed Higuchi model kinetics and had a diffusion-based release behavior; this shows that the drug has diffusion controlled release conditions afforded by the gel matrix. Ex vivo permeation experiments employing goat nasal mucosa demonstrated excessive permeation upon use of the in-situ gel base (78.6% over 8 h) than that of pure drug solution (46.2%), indicating the mucoadhesive and permeability increasing effects of the gel base [ 45 ]. The synergistic impact of Poloxamer 407 and 188, which momentarily changes the nasal epithelial barrier and allows drugs to be absorbed, can be viewed as the cause of the increased permeation. FTIR and DSC analyses revealed that there was no chemical interaction between zavegepant and formulation excipients, which means that there was stability of the drug in the formulation developed [ 46 ]. The finding of the drug has been stable within the accelerated conditions in three months and there was minimal difference in gelation temperature, PH and drug content further proving the robustness of the formulation. In pharmacodynamic analysis of the effect of the rat nitroglycerin induced-migraine model, the VF2 formulation was shown to have significant behavioral and biochemical recovery as it relates to the migraine control group. The treated animals with VF2 in the open field test recorded better locomotor results with time and by 4 hours after treatment, the results were recorded to be nearly normal. Equally, in light-dark box test, the VF2-treated animals took longer time in the light chamber suggesting that there was a decreased photophobia. In rats treated with VF2, the development of mechanical allodynia was reduced, which was confirmed by the von Frey filament test: the rats had a higher generated withdrawal threshold. The above behavioral changes indicate that VF2 was effective in treating the migraine symptoms caused by NTG [ 47 ]. The anti-migraine activities of the optimized formulation were also supported by bio-chemical tests. The migraine control group had higher values in nitric oxide and malondialdehyde (MDA) and CGRP, which are the highlight of migraine pathophysiology and indicate oxidative stress and trigeminovascular activation. On the other hand, in animals receiving VF2, there was a strong normalization of these markers [ 48 ]. The lowering of nitric oxide and MDA concentration proved the antioxidant property of zavegepant when administrated through the nose. The neuroprotective effect of VF2 was also supported by an increase in endogenous antioxidant enzymes like glutathione (GSH) and super oxide dismutase (SOD). Notably, VF2 CGRP levels were very much decreased compared to the migraine control group confirming the pharmacological importance of the CGRP receptor antagonism of zavegepant to abort a migraine attack. When compared to the zavegepant solution, VF2 formulation was found to be better in performance, and efficacy was found to be identical to that of the usual sumatriptan treatment. This implies that the thermoreversible nasal gel will not only guarantee the similar therapeutic effect but will also provide the benefit of the rapid non-invasive delivery method. In addition, the optimized formulation showed a prolonged rate of drug release, likely to prolong a relief of symptoms and limit requirements of the re-administration of drugs [ 49 ]. Repeated dosing of nasal mucosa by histopathologic studies demonstrated lack of inflammatory reaction, necrotic tissue and epithelial disturbance when using animals treated with VF2 establishing biocompatibility and non-consequentiality of the formulation, with no adverse effects on daily use of the product in the nasal administration route. This is a very important finding bearing in mind that there is a possibility of mucosal irritation when repeated drugs are administered like through the nose [ 50 ]. In general, the study was able to show that a thermoreversible in-situ nasal gel of zavegepant was an effective and new avenue of dealing with migraines. The formulation did not only enhance nasal residence time and bioavailability, but it also enabled specific brain delivery with minimal systemic adverse effect. The design incorporated QbD principles and enabled robustness, reproducibility, scalability of the formulation, which was critical in future clinical translation [ 51 ]. Conclusion This work succeeded to formulate and maximize the thermoresponsive, in-situ nasal fix of zavegepant as an active way of managing migraines. VF2 in the optimum formulation had an appropriate gelation temperature, viscosity, and desirable physicochemical characteristics of the administration of the nasal route of drug delivery. sustained slow release of drug with concomitant increased permeation through mucosa in vitro and ex vivo studies was seen when compared to the pure drug. Pharmacodynamic analysis in vivo of nitroglycerin induced migraine model revealed that locomotor activity, photophobia and mechanical allodynia improved in a significant manner in VF2. Biochemical examination showed less oxidation stress and CGRP, as well as, received antioxidant measures. The formulation was better than zavegepant solution and demonstrated similar efficacy to sumatriptan. The histopath evaluation showed that the safety in the nasal region was good. In general, the VF2 gel is a favorable, non-invasive, and successful intranasal modality of acute migraine delivery that needs the additional clinical and pharmacokinetic assessment. Abbreviations ANOVA: Analysis of Variance; FTIR: Fourier Transform Infrared Spectroscopy; UV: Ultraviolet Spectroscopy; DSC: Differential Scanning Calorimetry; QbD: Quality by Design; SD: Standard Deviation; MDA: Malondialdehyde; GSH: Glutathione; SOD: Superoxide Dismutase; NO: Nitric Oxide; CGRP: Calcitonin Gene-Related Peptide; NTG: Nitroglycerin; s.c.: Subcutaneous; VF2: Optimized Zavegepant Formulation; HPMC: Hydroxypropyl Methylcellulose; BCS: Biopharmaceutical Classification System; µL: Microliters; µmol: Micromole; nmol: Nanomole; U/mg: Units per milligram; mg/kg: Milligrams per kilogram. Declarations Conflict of interest Authors declare no conflict of interest regarding this study Author Contribution All Authors have equally contributed for research article Acknowledgement All changes have been made and updated manuscript had been attached. Data Availability No Database were genrated or analysed during current study References Safiri S, Pourfathi H, Eagan A, Mansournia MA, Khodayari MT, Sullman MJM, et al. Global, regional, and national burden of migraine in 204 countries and territories, 1990 to 2019. PAIN 2022;163:e293. https://doi.org/10.1097/j.pain.0000000000002275. Steiner TJ, Stovner LJ. Global epidemiology of migraine and its implications for public health and health policy. Nat Rev Neurol 2023;19:109–17. https://doi.org/10.1038/s41582-022-00763-1. Ashina M, Katsarava Z, Do TP, Buse DC, Pozo-Rosich P, Özge A, et al. Migraine: epidemiology and systems of care. The Lancet 2021;397:1485–95. https://doi.org/10.1016/S0140-6736(20)32160-7. Amiri P, Kazeminasab S, Nejadghaderi SA, Mohammadinasab R, Pourfathi H, Araj-Khodaei M, et al. Migraine: A Review on Its History, Global Epidemiology, Risk Factors, and Comorbidities. Front Neurol 2022;12. https://doi.org/10.3389/fneur.2021.800605. Peres MFP, Sacco S, Pozo-Rosich P, Tassorelli C, Ahmed F, Burstein R, et al. Migraine is the most disabling neurological disease among children and adolescents, and second after stroke among adults: A call to action. Cephalalgia 2024;44:03331024241267309. https://doi.org/10.1177/03331024241267309. Dong L, Dong W, Jin Y, Jiang Y, Li Z, Yu D. The Global Burden of Migraine: A 30-Year Trend Review and Future Projections by Age, Sex, Country, and Region. Pain Ther 2025;14:297–315. https://doi.org/10.1007/s40122-024-00690-7. Zhu Z, Tang Y, Li L, Ni H, Liu M, Chen Z, et al. The efficacy and safety of zavegepant nasal inhalation versus oral calcitonin-gene related peptide receptor antagonists in the acute treatment of migraine: a systematic review and network meta-analysis of the literature. J Headache Pain 2025;26:48. https://doi.org/10.1186/s10194-025-01984-7. Greco G, Monteith T. Intranasal zavegepant for the acute treatment of migraine. Expert Review of Neurotherapeutics 2024;24:1131–40. https://doi.org/10.1080/14737175.2024.2405741. Lipton RB, Croop R, Stock DA, Madonia J, Forshaw M, Lovegren M, et al. Safety, tolerability, and efficacy of zavegepant 10 mg nasal spray for the acute treatment of migraine in the USA: a phase 3, double-blind, randomised, placebo-controlled multicentre trial. The Lancet Neurology 2023;22:209–17. https://doi.org/10.1016/S1474-4422(22)00517-8. Pellesi L, Jedie B, Barhum F, Al-Abdullah S, Martelletti P, Xiao Z. Head-to-head relief: ubrogepant, rimegepant, and zavegepant in migraine treatment. Pain Management 2025;15:279–84. https://doi.org/10.1080/17581869.2025.2494494. Suhagiya K, Borkhataria CH, Gohil S, Manek RA, Patel KA, Patel NK, et al. Development of mucoadhesive in-situ nasal gel formulation for enhanced bioavailability and efficacy of rizatriptan in migraine treatment. Results in Chemistry 2023;6:101010. https://doi.org/10.1016/j.rechem.2023.101010. Karunakaran R, Nair SS, Pimpalshende PM, Devi D, Sachan K, Srivastav Y, et al. FORMULATION, OPTIMIZATION, AND CHARACTERIZATION OF A THERMOSENSITIVE IN SITU NASAL GEL INCORPORATING CHITOSAN NANOPARTICLES OF ZOLMITRIPTAN FOR THE MANAGEMENT OF ACUTE MIGRAINE ATTACKS n.d. Tanna V, Sawarkar SP, Ravikumar P. Exploring Nose to Brain Nano Delivery for Effective Management of Migraine. Current Drug Delivery 2023;20:144–57. https://doi.org/10.2174/1567201819666220401091632. Alshraim A, Alshora D, Ashri L, Alhusaini A, Alanazi N, Safwan NM. In Situ Thermosensitive Mucoadhesive Nasal Gel Containing Sumatriptan: In Vitro and Ex Vivo Evaluations. Polymers 2024;16:3422. https://doi.org/10.3390/polym16233422. Chattopadhyay S, Das S, Sarma KN. NOSE-TO-BRAIN DRUG DELIVERY: AN UPDATE TO THE ALTERNATIVE PATH TO SUCCESSFUL TARGETED ANTI-MIGRAINE DRUGS. Int J App Pharm 2021:67–75. https://doi.org/10.22159/ijap.2021v13i2.40404. Vidhi D, Patel P. Method development and Validation of UV Spectrophotometric estimation of Remogliflozin Etabonate in bulk and its tablet dosage form. RJPT 2021:2042–4. https://doi.org/10.52711/0974-360x.2021.00362. Gulhane CA, Panchale WA, Manwar JV, Bakal RL. Liquid Chromatographic Method for Simultaneous estimation of Thiocolchicoside and Etoricoxib from Tablet formulation. AJPA 2021:118–22. https://doi.org/10.52711/2231-5675.2021.00020. Zarghampour A, Jouyban K, Jouyban-Gharamaleki V, Jouyban A, Rahimpour E. A Description on the Shake-Flask and Laser Monitoring-Based Techniques for Determination of the Drug’s Solubility. Pharm Sci 2024;30:274–8. https://doi.org/10.34172/PS.2024.2. Rahimpour E, Moradi M, Sheikhi-Sovari A, Rezaei H, Rezaei H, Jouyban-Gharamaleki V, et al. Comparative Drug Solubility Studies Using Shake-Flask Versus a Laser-Based Robotic Method. AAPS PharmSciTech 2023;24:207. https://doi.org/10.1208/s12249-023-02667-9. Rojek B, Wesolowski M. A combined differential scanning calorimetry and thermogravimetry approach for the effective assessment of drug substance-excipient compatibility. J Therm Anal Calorim 2023;148:845–58. https://doi.org/10.1007/s10973-022-11849-9. Omari D, Sallam A, Al-Hmoud H, Rashid I. Modafinil-excipient compatibility study using differential scanning calorimetry. Journal of Advanced Pharmaceutical Technology & Research 2023;14:75. https://doi.org/10.4103/japtr.japtr_663_22. Bunaciu AA, Hoang VD, Aboul-Enein HY. Fourier transform infrared spectroscopy used in drug excipients compatibility studies. Applied Spectroscopy Reviews 2025;60:385–403. https://doi.org/10.1080/05704928.2024.2438747. Aminu N, Chan S-Y, Mumuni MA, Umar NM, Tanko N, Zauro SA, et al. Physicochemical compatibility studies of triclosan and flurbiprofen with excipients of pharmaceutical formulation using binary, ternary, and multi-combination approach. Futur J Pharm Sci 2021;7:148. https://doi.org/10.1186/s43094-021-00302-7. Hirun N, Kraisit P, Tantishaiyakul V. Thermosensitive Polymer Blend Composed of Poloxamer 407, Poloxamer 188 and Polycarbophil for the Use as Mucoadhesive In Situ Gel. Polymers 2022;14:1836. https://doi.org/10.3390/polym14091836. Chen Y, Lee J-H, Meng M, Cui N, Dai C-Y, Jia Q, et al. An Overview on Thermosensitive Oral Gel Based on Poloxamer 407. Materials 2021;14:4522. https://doi.org/10.3390/ma14164522. Chomchalao P, Saelim N, Lamlertthon S, Sisopa P, Tiyaboonchai W. A potential thermosensitive poloxamer 407-based in situ hydrogel containing moxifloxacin-loaded silk fibroin nanoparticles for prolonged ocular delivery. Journal of Drug Delivery Science and Technology 2024;99:105988. https://doi.org/10.1016/j.jddst.2024.105988. Mankar SD, Parjane SR, Siddheshwar SS, Dighe SB. Formulation, Optimization and In-Vivo Characterization of Thermosensitive In-Situ Nasal Gel Loaded with Bacoside a for Treatment of Epilepsy. AAPS PharmSciTech 2024;25:151. https://doi.org/10.1208/s12249-024-02870-2. Mathure D, Ranpise H, Awasthi R, Pawar A. Formulation and Characterization of Nanostructured Lipid Carriers of Rizatriptan Benzoate-Loaded In Situ Nasal Gel for Brain Targeting. ASSAY and Drug Development Technologies 2022;20:211–24. https://doi.org/10.1089/adt.2022.044. Alabdly A, Kassab HJ. Formulation Variables Effect on Gelation Temperature of Nefopam Hydrochloride intranasal in Situ Gel (Conference Paper) #. Iraqi Journal of Pharmaceutical Sciences 2022;31:32–44. https://doi.org/10.31351/vol31issSuppl.pp32-44. Salih OS, Ghareeb MM. Formulation and in-vitro evaluation of thermosensitive ciprofloxacin HCl in-situ gel for local nasal infection. IJDDT 2021;11:1295–301. Shaghlil L, Alshishani A, Sa’aleek AA, Abdelkader H, Al-ebini Y. Formulation and evaluation of nasal insert for nose-to-brain drug delivery of rivastigmine tartrate. Journal of Drug Delivery Science and Technology 2022;76:103736. https://doi.org/10.1016/j.jddst.2022.103736. Kammoun AK, Khedr A, Hegazy MA, Almalki AJ, Hosny KM, Abualsunun WA, et al. Formulation, optimization, and nephrotoxicity evaluation of an antifungal in situ nasal gel loaded with voriconazole‒clove oil transferosomal nanoparticles. Drug Delivery 2021;28:2229–40. https://doi.org/10.1080/10717544.2021.1992040. Uppuluri CT, Ravi PR, Dalvi AV, Shaikh SS, Kale SR. Piribedil loaded thermo-responsive nasal in situ gelling system for enhanced delivery to the brain: formulation optimization, physical characterization, and in vitro and in vivo evaluation. Drug Deliv and Transl Res 2021;11:909–26. https://doi.org/10.1007/s13346-020-00800-w. De Hoon I, Boukherroub R, De Smedt SC, Szunerits S, Sauvage F. In Vitro and Ex Vivo Models for Assessing Drug Permeation across the Cornea. Mol Pharmaceutics 2023;20:3298–319. https://doi.org/10.1021/acs.molpharmaceut.3c00195. Majid H, Bartel A, Burckhardt BB. Predictivity of standardized and controlled permeation studies: Ex vivo – In vitro – In vivo correlation for sublingual absorption of propranolol. European Journal of Pharmaceutics and Biopharmaceutics 2021;169:12–9. https://doi.org/10.1016/j.ejpb.2021.09.002. Yang D, Gao Y, Yin Z, Wang X, Meng X, Zou T, et al. Roxadustat alleviates nitroglycerin-induced migraine in mice by regulating HIF-1α/NF-κB/inflammation pathway. Acta Pharmacol Sin 2023;44:308–20. https://doi.org/10.1038/s41401-022-00941-3. Kang L, Tang W, Zhang Y, Zhang M, Liu J, Li Y, et al. The gut microbiome modulates nitroglycerin-induced migraine-related hyperalgesia in mice. Cephalalgia 2022;42:490–9. https://doi.org/10.1177/03331024211050036. Xu M, Zhang J, Shi Z, He Z, Zhao Y, Ling X, et al. Amelioration of nitroglycerin-induced migraine in mice via Wuzhuyu decoction: Inhibition of the MZF1/PGK1 pathway and activation of NRF2 antioxidant response. Journal of Ethnopharmacology 2024;326:117930. https://doi.org/10.1016/j.jep.2024.117930. Park S, Jung H, Han S-W, Lee S-H, Sohn J-H. Differences in Neuropathology between Nitroglycerin-Induced Mouse Models of Episodic and Chronic Migraine. International Journal of Molecular Sciences 2024;25:3706. https://doi.org/10.3390/ijms25073706. Wang L, Liu X, Zhu C, Wu S, Li Z, Jing L, et al. Environmental enrichment alleviates hyperalgesia by modulating central sensitization in a nitroglycerin-induced chronic migraine model of mice. J Headache Pain 2024;25:74. https://doi.org/10.1186/s10194-024-01779-2. Malviya V. Design and Characterization of Thermosensitive Mucoadhesive Nasal Gel for Meclizine Hydrochloride: Thermosensitive Nala Gel of Meclizine HCL. International Journal of Pharmaceutical Sciences and Nanotechnology(IJPSN) 2022;15:5782–93. https://doi.org/10.37285/ijpsn.2022.15.1.6. Trupa K, Gandhi S, Nidhi P, Dhruvisha C, Kadiya MJ. FORMULATION AND EVALUATION NASAL IN SITUGEL OF RIZATRIPTAN. International Journal of Pharmacy Research & Technology (IJPRT) 2019;9:49–64. https://doi.org/10.31838/ijprt/09.02.03. Marzouk MA, Osman DA, Abd El-Fattah AI. Formulation and in vitro evaluation of a thermoreversible mucoadhesive nasal gel of itopride hydrochloride. Drug Development and Industrial Pharmacy 2018;44:1857–67. https://doi.org/10.1080/03639045.2018.1504059. Jaiswal J, Anantvar SP, Narkhede MR, Gore SV, Mehta K. Formulation and evaluation of thermoreversible in-situ nasal gel of metoprolol succinate. International Journal of Pharmacy and Pharmaceutical Sciences 2012;4:96–102. Altuntaş E, Yener G. Formulation and Evaluation of Thermoreversible In Situ Nasal Gels Containing Mometasone Furoate for Allergic Rhinitis. AAPS PharmSciTech 2017;18:2673–82. https://doi.org/10.1208/s12249-017-0747-8. Bai L, Lei F, Luo R, Fei Q, Zheng Z, He N, et al. Development of a Thermosensitive In-Situ Gel Formulations of Vancomycin Hydrochloride: Design, Preparation, In Vitro and In Vivo Evaluation. Journal of Pharmaceutical Sciences 2022;111:2552–61. https://doi.org/10.1016/j.xphs.2022.04.011. Raval M, Bagada H. Formulation and Evaluation of Cyclodextrin-Based Thermosensitive In Situ Gel of Azithromycin for Periodontal Delivery. J Pharm Innov 2021;16:67–84. https://doi.org/10.1007/s12247-019-09422-3. Zhao Y, Wang T, Chen Z, Ren H, Song P, Zhu Y, et al. Development and Evaluation of a Thermosensitive In Situ Gel Formulation for Intravaginal Delivery of Lactobacillus gasseri. Pharmaceutics 2022;14:1934. https://doi.org/10.3390/pharmaceutics14091934. Gugleva V, Michailova V, Mihaylova R, Momekov G, Zaharieva MM, Najdenski H, et al. Formulation and Evaluation of Hybrid Niosomal In Situ Gel for Intravesical Co-Delivery of Curcumin and Gentamicin Sulfate. Pharmaceutics 2022;14:747. https://doi.org/10.3390/pharmaceutics14040747. Polat HK, Arslan A, Ünal S, Haydar MK, Aytekin E, Gözcü S, et al. Formulation Development of Dual Drug-Loaded Thermosensitive Ocular In Situ Gel Using Factorial Design. J Pharm Innov 2023;18:768–88. https://doi.org/10.1007/s12247-023-09762-1. Salem HF, Ali AA, Rabea YK, El-Ela FIA, Khallaf RA. Glycerosomal thermosensitive in situ gel of duloxetine HCl as a novel nanoplatform for rectal delivery: in vitro optimization and in vivo appraisal. Drug Deliv and Transl Res 2022;12:3083–103. https://doi.org/10.1007/s13346-022-01172-z. Additional Declarations No competing interests reported. Supplementary Files GraphicalAbstract.png Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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11:17:01","extension":"xml","order_by":28,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":236993,"visible":true,"origin":"","legend":"","description":"","filename":"6ce7795986e2475088da65efd4e071041structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/dbce2d53ae0ddf4d160311ad.xml"},{"id":92168388,"identity":"83232b82-e392-4a9a-ac23-a2cec9eae025","added_by":"auto","created_at":"2025-09-25 11:25:01","extension":"html","order_by":29,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":254688,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/f95f8c20fe7a18447a0f3244.html"},{"id":92167400,"identity":"7ee02e80-54e6-4322-8ad0-b012a8737f1c","added_by":"auto","created_at":"2025-09-25 11:17:00","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":21155,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eStructure of Zavegepant\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/fc74e04caac7d941b8586406.png"},{"id":92167399,"identity":"d040e483-ab5d-45b7-bc98-29a13cc52f7d","added_by":"auto","created_at":"2025-09-25 11:17:00","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":748760,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMucoadhesive strength determination by modified balance method\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/c653c033280dd96dbaf4a762.png"},{"id":92168376,"identity":"e824fcad-771a-4748-9132-6f2456ca2b4c","added_by":"auto","created_at":"2025-09-25 11:25:00","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":52332,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCalibration curve of zavegepant\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/7e150c4848e49e35778a2a44.png"},{"id":92168378,"identity":"98bc9dcd-cb84-466b-976b-2fb1a387d48b","added_by":"auto","created_at":"2025-09-25 11:25:00","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":207261,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDSC thermograms of (A) pure zavegepant (peak at 264.07°C) and (B) physical mixture (peaks at 262.13°C and 275.50°C)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/842bf4ec2a86a5a70771cb67.png"},{"id":92168383,"identity":"c19b31ae-24b4-408a-ada2-4eb6d19f1ea2","added_by":"auto","created_at":"2025-09-25 11:25:00","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":170145,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFTIR spectra of (A) zavegepant (B) physical mixture (Drug + Excipients)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/237f63f202ec57834ce67e95.png"},{"id":92167410,"identity":"b15e5278-8aa5-41f3-9b70-ad68b30f78e9","added_by":"auto","created_at":"2025-09-25 11:17:00","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":264420,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eImage represents \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein-situ\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003enasal formulation in sol to gel state\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/3cdf8c747f80f60999da0a97.png"},{"id":92168752,"identity":"6e8a9f4e-24b4-4d15-a905-fb78d76fe457","added_by":"auto","created_at":"2025-09-25 11:33:00","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":389039,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eContour plots and 3D response surface plots showing the effect of Pluronic F-127 (A) and xanthan gum (B) on gelation temperature (°C) and mucoadhesive strength (dyn/cm²)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/c46d298ed4d693eaa5d73a0f.png"},{"id":92167404,"identity":"b341ce65-5349-4f92-9e02-3737f781f9a3","added_by":"auto","created_at":"2025-09-25 11:17:00","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":61055,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eEx vivo\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e drug permeation from thermosensitive \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein situ\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e nasal gel\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/56dc887636dddb18d432bc70.png"},{"id":92167406,"identity":"d04566ed-fcb9-47db-b580-c636b6a27fb8","added_by":"auto","created_at":"2025-09-25 11:17:00","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":122993,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of Different Formulations on Behavioral Parameters in Nitroglycerin-Induced Migraine Model\u003c/strong\u003e \u003cstrong\u003eThe graphs depict changes in (A) locomotor activity, (B) photophobia, and (C) mechanical allodynia over 4 hours post-NTG administration. Group II (migraine control) showed significant behavioral deficits, while Group III (sumatriptan) and Groups IV–V (zavegepant-treated) demonstrated progressive improvements. The optimized in-situ nasal gel (VF2) showed superior efficacy compared to plain zavegepant solution. Values represent mean ± SD (n = 6).\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/5e8057d08ac38e6011dab29b.png"},{"id":92167427,"identity":"79d22f4d-d8d9-48b1-ae46-843cccd03338","added_by":"auto","created_at":"2025-09-25 11:17:01","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":705039,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eInduction of Migraine and Intranasal Administration of Zavegepant Gel in Wistar Rats\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/df8587db2f208162c5bfd77c.png"},{"id":92184181,"identity":"105a501a-2434-44ef-8ff8-b5f48ca368c2","added_by":"auto","created_at":"2025-09-25 14:02:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6352796,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/f42f6601-2a3e-43b9-bd15-a074208fe7a9.pdf"},{"id":92168751,"identity":"26488336-f3b7-4a63-919a-b64e11ddaa6c","added_by":"auto","created_at":"2025-09-25 11:33:00","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":310829,"visible":true,"origin":"","legend":"","description":"","filename":"GraphicalAbstract.png","url":"https://assets-eu.researchsquare.com/files/rs-7281614/v1/57a3131e7254db4e82642ce6.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eDevelopment and Optimization of Thermoreversible in-situ Nasal Gel Loaded With Zavegepant for Treatment of Migraine\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eMigraine is a significant health issue in the world, with a world prevalence level of migraine disability of about 14% of all the world population, with higher risk in a group of population where women are more vulnerable at the rate of 18% and men 9% [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. As a neurological disorder, it has a massive economic cost to the United States, of around \u003cspan\u003e$\u003c/span\u003e27\u0026nbsp;billion a year in direct care costs and productivity loss [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The existing methods of treatment are not fully effective, as the relief provided by the current drugs fails to cope with the necessity in approximately 40% of patients. Other limitations included in these factors are delayed onset of action, large first-pass metabolism, and GI disorders which are relatively problematic during nausea experienced with migraines. Treatment gap is especially high in the middle and low-income countries where proportion of migraine patients who obtain proper treatment is less than 7\u0026ndash;14% [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Migraine is also given by periodical attacks of moderate to severe headache, which is always combined with the presence of nausea and vomiting, photosensitivity (sensitivity to light), and phonophobia (sensitivity to sound) [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. There are numerous triggers of migraine although it is not clearly understood; these triggers occur in the form of genetic predisposition, environmental factors, hormonal shifts and dysfunction of the brain [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The pathophysiology of migraines has a great dependence on the activation of the trigeminovascular system that subsequently releases neuropeptides such as calcitonin gene-related peptide (CGRP). This leads to vasodilation, neurogenic inflammation and intensification of the nerve endings pain response. Migraines with aura have also been attributed to cortical spreading depression, a neuronal depolarization wave through the cerebral cortex [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eZavegepant is a new calcitonin gene-related peptide (CGRP) receptor antagonist in the acute treatment of migraine structure of zavegepant is depicted in Fig.\u0026nbsp;1 [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. It is the first and only CGRP antagonist as a nasal spray formulation that can be used to obtain immediate relief to symptoms of migraine [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Zavegepant has an advantage of quick absorption via the nose since oral CGRP inhibitors take time to react against nausea and vomiting in migraine patients [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The mechanism of action of the zavegepant drug is inhibition of CGRP receptors and prevention of vasodilation and neuroinflammation of migraine pathophysiology. Recent clinical trials indicate that, zavegepant can initially relieve pain and symptom-free in two hours and has few safety or tolerability concerns [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eOne of the potentially successful systems to deliver the drug treating migraine is thermosensitive in situ gels applied to the nasal route [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The formulations are in liquid form at the room temperature and change to gel when they are in contact with nasal mucosa at a body temperature. This peculiarity leads to better retention of drugs to the location of drug absorption and better bioavailability and therapeutic effect [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The most widely used thermosensitive polymer is poloxamer 407 (Pluronic F-127), which is a nonionic triblock copolymer synthesised using polyethylene oxide (PEO) and polypropylene oxide (PPO) [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Poloxamer 407 is temperature-responsive to form gels and therefore this properly gels at physiological temperature leaving the drug to stay long in the nasal cavity [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Nasal route provides the multiple advantages such as rapid absorption, ease of administration, avoids first pass and etc. Moreover, the drugs can enter through the central nervous system via the olfactory and trigeminal nerve routes. Thermosensitive in situ nasal gels are specifically appealing to the treatment of acute migraine attacks because of a need of a sweeping speed of action [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThis is the major goal of the study; to invent and perfect a thermoreversible in situ nasal gel system that incorporates zavegepant to treat migraines. The pursuit of this study is to improve the bioavailability and residence time of the drug in the nasal cavity by using a unique system of gel that can exist in liquid state at room temperature but become gel at the temperature existing within the nasal cavity. This new method of drug delivery will enhance adherence in the patient, results in the fast onset of action, and continuous slow release of a drug combined with the avoidance of first-pass metabolism and the direct delivery of zavegepant via nose to the brain.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e\u003cstrong\u003eMaterials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZavegepant (\u0026ge;98% purity) was procured from Pfizer Inc. (New York, USA). Pluronic F-127 (pharmaceutical grade) and xanthan gum (food grade, particle size \u0026lt;150 \u0026mu;m) were sourced from Research Lab Fine Chem Industries (Mumbai, India). Polyethylene glycol 400 (PEG 400, pharmaceutical grade) and Benzalkonium chloride (0.01% w/v, preservative grade) were purchased from Merck Life Science Pvt. Ltd. (Mumbai, India). Nitroglycerin solution (5 mg/mL injection) was obtained from Sun Pharmaceutical Industries Ltd. (Mumbai, India). Sumatriptan succinate (\u0026gt;99% purity), HPLC-grade acetonitrile, methanol, and water were purchased from Sigma-Aldrich (Mumbai, India). Potassium bromide (KBr) (spectroscopic grade) for FTIR analysis was supplied by Himedia Laboratories (Mumbai, India). All other chemicals and reagents used were of analytical grade and used without further purification.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMETHODS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCalibration Curve of Zavegepant\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUV-visible spectrophotometer (Shimadzu UV-1800, Shimadzu India, Mumbai) was used to prepare a calibration curve of zavegepant. Zavegepant (10 mg) was reconstituted in 100 mL methanol to produce a stock solution of 100 1259038 g/mL. Standard solutions of 5, 10, 15, 20, 25 and 30 0g/ml were prepared by proper dilution with methanol. The solutions were measured as Absorbance at 239 nm \u0026lambda;\u003csub\u003emax\u003c/sub\u003e against methanol blank. The plot of the absorbance with respect to the concentration was used to form the calibration curve that was used to get the linear regression equation. Correlation coefficient (R\u003csup\u003e2\u003c/sup\u003e), slope and y- intercept were used to validate the linearity. Percent standard deviation of response as well as slope was used in calculating LOD and LOQ. Each measurement was done in triplicate (n=3) [16,17].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSolubility Study\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe saturation shake flask method was used to determine the solubility of zavegepant in five solvents such as methanol, ethanol, distilled water, DMSO, and phosphate buffer at pH 6.8. Surplus quantities of the drug (10 mg) were placed in 10 mL of each of solvents in closed vials. A 24 hour orbital shaking at 100 rpm was carried at 25 \u0026plusmn; 2\u0026deg;C in orbital incubator (REMI CIS-24 BL, India). Once in equilibrium, the samples were centrifuged at 5000 rpm and after 15 minutes filtration was performed using 0.45 mm membrane filter. To mix, the filtrate was diluted properly with methanol and analyzed through the UV-visible spectrophotometer at 239 nm. Each of the experiments was repeated three times (n=3) and the means of solubility values were reported as mean\u0026plusmn;SD [18,19].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDifferential Scanning Calorimetry (DSC)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe thermal properties of zavegepant and possible drug-excipient interaction were studied using differential scanning calorimetry. Analysis was done through a DSC-60 Plus (Shimadzu Corporation, Kyoto, Japan). Pure zavegepant and physical mixture were weighed accurately (5 mg) and placed in standard aluminum pans and heated between 30-300\u003csup\u003eo\u003c/sup\u003eC with heating rate of 10\u003csup\u003e\u0026nbsp;o\u003c/sup\u003eC/min with nitrogen atmosphere (50 mL/min flow rate). A reference was an empty sealed aluminum pan. The alteration of endothermic peak characteristics was tested to determine the interaction of drug-excipient and alteration of crystallinity structure. All the measurements were done three-fold (n=3) [20,21].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFourier Transform Infrared (FTIR) Spectroscopy\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAnalysis of chemical interactions between zavegepant and formulation components Chemical interactions between zavegepant and formulation components were examined by FTIR spectroscopy using FTIR spectrophotometer (IRAffinity-1S, Shimadzu Corporation, Kyoto, Japan) with DTGS detector. Pure zavegepant, single polymers, and physical mixtures were prepared by separately mixing 2 mg of each sample with 100 mg dry KBr and pressed into their transparent pellets using a hydraulic pressure in a vacuum. Spectra in the range of 4000-400 cm\u003csup\u003e-1\u003c/sup\u003e were obtained with a resolution of 4 cm\u003csup\u003e-1\u003c/sup\u003e. This measurement was done in triplicates (n=3). Frequencies of peak shift, vanishing, or broadening patterns were beforehand revealed in order to identify drug-excipient interactions [22,23].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eOptimization of In-situ Nasal Gel\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA 3\u0026sup2; full factorial design was employed to systematically investigate the influence of two critical formulation variables: Pluronic F-127 (X₁), a thermosensitive gelling agent, and xanthan gum (X₂), a mucoadhesive polymer, on the performance of the in-situ nasal gel. Each factor was evaluated at three levels, resulting in nine experimental runs. The independent variables (X₁ and X₂) were assessed for their effects on two key dependent responses: gelation temperature (Y₁) and mucoadhesive strength (Y₂). The design layout, including both actual and coded values, is presented in Tables 1 and 2.\u003c/p\u003e\n\u003cp\u003eThe relationship between formulation variables and responses was fitted to a second-order polynomial equation:\u003c/p\u003e\n\u003cp\u003eY = b₀ + b₁X₁ + b₂X₂ + b₁₂X₁X₂ + b₁₁X₁\u0026sup2; + b₂₂X₂\u0026sup2;\u003c/p\u003e\n\u003cp\u003ewhere Y denotes the predicted response, X₁ and X₂ are the coded levels of independent variables, and coefficients represent the magnitude and direction of influence. Analysis of variance (ANOVA) was performed to determine statistical significance (p\u0026lt;0.05). The optimal formulation was identified using desirability function with constraints set as gelation temperature (32-36\u0026deg;C) and maximized mucoadhesive strength [24,25].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1: Design Matrix of 3\u0026sup2; Factorial Study Highlighting Independent Factors, Coded Levels, Dependent Measures, and Optimization Targets\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFactors (Independent Variables)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" style=\"width: 51px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLevels\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLow (-1)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMedium (0)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHigh (+1)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003eA: Pluronic F-127 (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003eB: Xanthan gum (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDependent Variables\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGoal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" style=\"width: 83px;\"\u003e\n \u003cp\u003eY₁: Gelation Temperature (\u0026deg;C)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003e30-34\u0026deg;C\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" style=\"width: 83px;\"\u003e\n \u003cp\u003eY₂: Mucoadhesive Strength (N/cm\u0026sup2;)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eMaximize\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2: Ingredient Profile of Zavegepant-Incorporated In-Situ Nasal Gel Formulations(VF1-VF9)\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eIngredients\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eZavegepant (mg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePluronic F-127 (%w/v)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eXanthan gum (%w/v)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBenzalkonium chloride (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePEG 400 (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eDistilled water\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eq.s. to 10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of Zavegepant-loaded \u003cem\u003eIn-situ\u003c/em\u003e Nasal Gel\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe cold method was used in the preparation of the in-situ nasal gel. Pre-cooled phosphate buffer (pH 6.4 and 4 C) was gradually added to Pluronic F-127 and this was stirred continuously at 400 rpm (Remi Equipment Pvt. Ltd., Mumbai, India) The dispersion was re-frigerated-after full hydration-at 4\u0026plusmn;1\u003csup\u003eo\u003c/sup\u003eC, over a 24 hours period. Xanthan gum was dispersed individually into phosphate buffer and eventually put in Pluronic solution continuously till the mixture was stirred. The dissolved solution of Zavegepant (10 mg) was mixed into the PEG 400 and polymer solution. Benzalkonium chloride (0.01%) was added as preservative. To complete the final volume, phosphate buffer was added up to 10 mL and the samples were stored at 4\u0026plusmn;1\u003csup\u003eo\u003c/sup\u003eC within 24 hours and then evaluated. Each formulation was prepared in three replicates (n=3) and the pH was set between 6.4\u0026plusmn;0.2 [26].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEvaluation of Thermosensitive In-situ Nasal Gel\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003epH Determination\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn order to make sure that the pH in situ nasal gel needed to be suitable to apply in the nasal system, a digital pH meter (Eutech pH 700, Singapore) was used. 1 g of the formulation was dispersed in 10 mL of distilled water and the electrode inserted to measure the pH when it reached its normal level. pH 4.0, pH 7.0 and pH 9.2 buffer solutions of standard concentration were used to calibrate the instrument. Measurements were made three times (n=3) at 25\u0026plusmn;2\u003csup\u003eo\u003c/sup\u003eC [27].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGelation Temperature\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe tube inversion method of determining gelation temperature was used. A water bath temperature was pre-set to 20\u003csup\u003eo\u003c/sup\u003eC by placing a 5 mL formulation in a test tube. The temperature was raised at a rate of 1\u003csup\u003eo\u003c/sup\u003eC/minute with calibrated thermometers being used to track this process. The test tube was turned upside down at 90\u003csup\u003eo\u0026nbsp;\u003c/sup\u003eangle at every temperature increment and observation of the formation of gel was made. The point at which the formulation ceased to flow has been noted as the gelation temperature. Each measurement was done three times (n=3) [28].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGelation Time\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGelation time was recorded in a slightly altered method to that of Miller and Donovan method. An amount of 2 mL of gel formulation was placed in 1.0 cm (diameter) test tube and sealed with Parafilm, and vertical setting in a circulating water bath at 37\u003csup\u003eo\u003c/sup\u003eC temperature. Following a 10-minutes equilibration period, the tube was placed in a horizontal position to note the formation of gels. The sol-to-gel transition was found at the moment when the formulation ceased to flow. The experimental operations were repeated thrice (n=3) [29].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eViscosity Measurement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMeasures of viscosity of the in-situ nasal gel pre and post gelation were grabbed using a Brookfield viscometer (LVDV-E, Brookfield Engineering Labs) where spindle No. 64 (Technosys Instruments, Thane, India) was used. The 10 mL formulation was used to measure at 25\u0026plusmn;0.5\u003csup\u003eo\u003c/sup\u003eC and 34 \u0026plusmn; 0.5\u0026deg;C. During the test, the spindle speed was kept at 50 rpm. Measurement was carried out after equilibrating in a 34 \u0026plusmn; 1\u0026deg;C water bath at 20 minutes after gelation. The values of viscosity were measured in centipoise (cP). All the measurements were carried out three times (n=3) [30].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSpreadability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEngineering variables were calculated by slip and drag method to determine the spreadability. Clean glass slides (7.5 x 2.5 cm) were used to put the properly weighed gel in between them. A 100 g weight was pressed on the upper slide to make it evenly distributed by 5 minutes. A wooden block was used to support the lower slide. The automatic separation of the upper slide following the removal of the weights (t) was recorded. Based on the following formula, spreadability (S) was calculated:\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eS = (M \u0026times; L)/T\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ewhere M is applied weight (g), L is slide length (cm), and T is time (sec). All measurements were performed in triplicate (n=3) [31].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDrug Content Uniformity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDrug content was analyzed as: gel (equivalent to 1 mg zavegepant) was dissolved in 10 mL of methanol with vortexing at 5 minutes. The solution was diluted and then filtered using 0.45 \u0026mu;m membrane filter. UV-visible spectrophotometer was used to examine absorbance in 239 nm, with blank consisting of methanol. The concentration of drugs would be determined based on the calibration curve already obtained. Each of the tests was repeated thrice (n=3) [32].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMucoadhesive Strength\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe modified balance (Figure 2)\u0026nbsp;technique was adopted to test the mucoadhesive strength on new excised goat nasal mucus. Mucosal tissues of 1 cm 2 in size were taken and were washed using distilled water and phosphate buffer (pH 6.4) to flake out the connective tissue. The mucosa including the mucosal side were attached to glass with cyanoacrylate adhesive. The support was put in phosphate buffer solution on the right hand pan of a physical balance. The weight placed at the left pan was 5 g in order to get the equilibrium. Gel was used as 1g and it was used to completely cover the mucosa and this was done in 2min. Gradually water was added to the left pan till detachment of mucosa occurred in the gel. Mucoadhesive strength was measured in terms of the weight of detachment, and converted in number of dyne/cm\u003csup\u003e2\u003c/sup\u003e. The experiments were done thrice (n=3) [33].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eEx Vivo\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;Permeation Study\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFranz diffusion cells were used in permeation studies where goat nasal mucosa was used. The mucosal tissue after cleaning was evenly sliced and placed between donor and receptor chambers and the mucosal surface exposed toward the donor. The receptor compartment was filled with 20 mL of phosphate buffer (pH 6.4) that was kept at 37\u0026plusmn;0.5\u003csup\u003eo\u003c/sup\u003eC and stirred continuously at 100 rpm. The donor chamber was filled with gel formulation 1 g. Aliquots (1 mL) of the receptor medium were removed at specific times (0, 1, 2, 3, 4, 5, 6, 7 and 8 hours) and replaced with fresh buffer to ensure sink conditions. The samples were filtered by 0.45 2003 um membrane filter and UV-visible spectrophotometer was used to analyze with reference to 239 nm. The Jss and the Kp were determined based on the linear part of the permeation curve. Each experiment was done with triplicates (n=3) [34,35].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAnimals and Ethical Clearance\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe male Wistar rats were maintained in polypropylene cages under controlled of temperature (25\u0026plusmn;2 \u003csup\u003eo\u003c/sup\u003eC), relative humidity (60\u0026plusmn;5 percent)) in standard laboratory conditions 12:12 light/dark. Food was standard pellet diet and ad libitum water. The study was done after one week of acclimatization. All animal studies were carried out on the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) and approved by the Institutional Animal Ethics Committee (IAEC) (Approval No. MESCOP/IAEC/2023-24/01). Proper steps were taken that would ensure the minimum suffering of animals and the least number of animals are utilized [36].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNitroglycerin (NTG)-Induced Migraine Model\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNitroglycerin (NTG)-induced migraine model in rats was used, because the developed zavegepant-loaded in-situ nasal gel had been tested on a validated rat model of migraine before. It was found that NTG solution (5 mg/kg, subcutaneously, injected in the dorsal region) caused migraine-like symptoms. In post-NTG animals, the trends of commonly observed migraine behaviour were simulated such as a motor dysfunction, photophobia, phonophobia and allodynia on mechanical stimulus. The results of these responses were quantified by common accepted conductions of behaviors: a locomoter activity was assessed through the open field test, photo phobia was experienced through the light dark box test as well as mechanical allodynia through the von frey filament approach. Symptoms akin to migraine usually appeared in 1 hour after injection of NTG and thereafter interventions were taken to treat it in accordance with the experiment design [37,38].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental Design\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe animals were randomized into five groups containing 6 animals each (n = 6) and evaluated to assess the anti-migraine activity of: Group I (Normal Control): intravenously administered saline; Group II (Migraine Control): intravenously given nitroglycerin (NTG, 5 mg/kg, s.c.) with no treatment; Group III (Standard Treatment): NTG (5 mg/kg, s.c.) given with an intranasal dose of sumatriptan (6 mg/kg); Group IV given NTG (5 mg/kg, In the case of intranasal administration, 20 \u0026mu;L of respective formulation was administered into each nostril with a micropipette equipped with a low-density polyethylene tube in the animal supine position under mild (2-3% of isoflurane) anesthesia line. The behavioral tests were made at 0 hour (baseline), 1 hour (post-NTG, pre-treatment), 2, 3 and 4 hours after injection of NTG. The experimentation ended with the death of animals by exposing them to CO\u003csub\u003e2\u003c/sub\u003e inhalation, and the brain tissues were harvested in order to be checked on biochemical and pathological level [39,40].\u003c/p\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cp\u003e\u003cstrong\u003eCalibration Curve of Zavegepant\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe calibration plot of zavegepant was a straight line with correlation coefficient (R 2 ) of 0.9991 in the range of 5-30 \u0026mu;g/mL. The quantitative analysis showed strong enlightenment of good linearity and the ability of the equation Y = 0.0182x + 0.0091 to support quantitative analysis (Figure 3). It was established that limit of detection (LOD) and limit of quantification (LOQ) was 0.15 \u0026mu;g/mL and 0.45 \u0026mu;g/mL, respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSolubility Study\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSolubility of Zavegepant in various solvents was largely different. The maximum solubility was seen in DMSO (112.8\u0026plusmn;3.45 mg/mL), methanol (45.6\u0026plusmn;1.02 mg/mL), ethanol (12.5\u0026plusmn;0.43 mg/mL), phosphate buffer pH 6.8 (4.3\u0026plusmn;0.21 mg/mL), and water (0.06\u0026plusmn;0.01 mg/mL). Zavegepant was freely soluble in DMSO, soluble in methanol, sparingly soluble in ethanol, slightly soluble in the phosphate buffer and practically insoluble in the water as per the Indian Pharmacopoeia classification (Table 3).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3: Solubility study of zavegepant in different solvents\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSr. No.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSolvent\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSolubility (mg/mL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eClassification\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33px;\"\u003e\n \u003cp\u003eWater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e0.06 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003ePractically insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33px;\"\u003e\n \u003cp\u003eEthanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e12.5 \u0026plusmn; 0.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003eSparingly soluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33px;\"\u003e\n \u003cp\u003eMethanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e45.6 \u0026plusmn; 1.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003eSoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33px;\"\u003e\n \u003cp\u003ePhosphate Buffer pH 6.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e4.3 \u0026plusmn; 0.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003eSlightly soluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33px;\"\u003e\n \u003cp\u003eDMSO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e112.8 \u0026plusmn; 3.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003eFreely soluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003eValues expressed as mean \u0026plusmn; SD (n=3)\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDifferential Scanning Calorimetry (DSC)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePure zavegepant showed a sharp endothermic peak at 264.07\u003csup\u003eo\u003c/sup\u003eC, which showed that it is crystalline. Here the physical mixture had two endothermic peaks located at 262.13\u003csup\u003eo\u003c/sup\u003eC and 275.5\u003csup\u003eo\u003c/sup\u003eC which represented zavegepant and excipients respectively (Figure 4). The fact that no major peak shifts, broadening or disappearance occurred was indicative that there was no significant physicochemical interaction between the drug and excipients which implied close compatibility with formulation development.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFTIR Spectroscopy\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePure zavegepant revealed the characteristic FTIR spectra of 3281 cm\u003csup\u003e-1\u003c/sup\u003e (N-H stretching), 2934.3 and 2850.5 cm\u003csup\u003e-1\u003c/sup\u003e (C-H stretching), 1680.0 cm\u003csup\u003e-1\u003c/sup\u003e (C=O stretching) as well as 1608.5 cm\u003csup\u003e-1\u003c/sup\u003e (aromatic C=C stretching). Peaks observed on physical mixture were comparable at 3310.8 cm\u003csup\u003e-1\u003c/sup\u003e (N-H), 2932.0\u003csup\u003e-1\u003c/sup\u003e and 2857.7cm\u003csup\u003e-1\u003c/sup\u003e (C-H), 1689.9\u003csup\u003e-1\u003c/sup\u003e and 1724.0 cm\u003csup\u003e-1\u003c/sup\u003e (C=O) and 1608.0 cm\u003csup\u003e-1\u003c/sup\u003e (C=C) (Figure 5, Table 4) with no drastic changes. This ascertained the non-existence of a chemical interaction between zavegepant and excipients.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4:\u003c/strong\u003e \u003cstrong\u003eFTIR Interpretation of Pure Zavegepant and Physical Mixture\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eFunctional Group\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eStandard Wavenumber (cm⁻\u0026sup1;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eObserved in Pure Drug (cm⁻\u0026sup1;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eObserved in Physical Mixture (cm⁻\u0026sup1;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eN-H stretching\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3300-3400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3281\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3310.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eC-H stretching\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2850-2950\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2934.3, 2850.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2932.0, 2857.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eC=O stretching\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1650-1750\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1680.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1689.9, 1724.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAromatic C=C stretch\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1580-1620\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1608.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1608.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eC-N stretching\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1200-1350\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1245.3, 1280.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1240.6, 1281.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eC-O stretching\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1000-1300\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1132.7, 1024.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1131.7, 1027.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003ePhysicochemical Evaluation of Thermoresponsive In-Situ Nasal Gel\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the nine formulations (VF1-VF9) had clear appearance, homogeneous, and good consistency, which indicated successful drug incorporation and polymer dispersion. The physical\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePhysical Characterization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe uniform appearance and a transparent homogeneous texture indicate the successful placement of drugs and dispersion of polymer in all the nine formulations (VF1-VF9) as well as their excellent consistency. This compliances with the formulations suitability in nasal delivery as revealed by physical stability and reproducibility between any two different batches (Table 5).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5:\u003c/strong\u003e \u003cstrong\u003ePhysical characterization of thermosensitive in-situ nasal gel formulations\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFormulation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAppearance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHomogeneity\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConsistency\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePhase Separation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eVF9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eTransparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eHomogeneous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eExcellent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAll physicochemical characteristics of formulations showed an appropriate profile of nasal delivery. pH values of the formulations were between 5.1 and 6.4, confirming their suitability as nasal delivery. The degree of spreadability was 7.6 \u0026plusmn;0.76 to 11.6\u0026plusmn;0.78 cm which is good. The viscosity of all gels before the gelation point ranged between 170.25\u0026plusmn;1.63 and 284.86\u0026plusmn;1.77 cP as established in the charts of the viscosity obtained, and after the gelation point increased significantly to 11638.1\u0026plusmn;132.2 and 12342.8\u0026plusmn;102.4 to substantiate their effective thermosensitive behavior. Content uniformity of drugs was high in all formulations (94.42 to 9877%) and shows good reproducibility of manufacturing, as shown in Table 6.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6:\u003c/strong\u003e \u003cstrong\u003ePhysicochemical characteristics of zavegepant-loaded in-situ gel formulations\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eFormulation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003epH\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpreadability (cm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eViscosity before gelation (cP)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eViscosity after gelation (cP)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrug content (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.5 \u0026plusmn; 0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e8.2 \u0026plusmn; 0.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e170.25 \u0026plusmn; 1.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11782.5 \u0026plusmn; 128.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e94.42 \u0026plusmn; 1.23\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.8 \u0026plusmn; 0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e7.6 \u0026plusmn; 0.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e187.91 \u0026plusmn; 0.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11968.4 \u0026plusmn; 115.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e97.32 \u0026plusmn; 0.89\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.2 \u0026plusmn; 0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e8.8 \u0026plusmn; 0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e220.45 \u0026plusmn; 1.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e12092.7 \u0026plusmn; 107.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e97.67 \u0026plusmn; 0.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.3 \u0026plusmn; 0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e9.1 \u0026plusmn; 0.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e218.22 \u0026plusmn; 2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11638.1 \u0026plusmn; 132.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e96.56 \u0026plusmn; 1.04\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.1 \u0026plusmn; 0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11.6 \u0026plusmn; 0.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e192.32 \u0026plusmn; 1.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11864.3 \u0026plusmn; 121.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e94.86 \u0026plusmn; 1.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.1 \u0026plusmn; 0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e8.3 \u0026plusmn; 0.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e220.15 \u0026plusmn; 2.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e12125.6 \u0026plusmn; 110.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e97.45 \u0026plusmn; 0.92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.4 \u0026plusmn; 0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e10.2 \u0026plusmn; 0.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e284.86 \u0026plusmn; 1.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e12342.8 \u0026plusmn; 102.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e98.74 \u0026plusmn; 0.65\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.5 \u0026plusmn; 0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e9.4 \u0026plusmn; 0.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e246.12 \u0026plusmn; 0.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e12210.5 \u0026plusmn; 108.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e98.77 \u0026plusmn; 0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.4 \u0026plusmn; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e8.7 \u0026plusmn; 0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e268.51 \u0026plusmn; 0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e12184.9 \u0026plusmn; 113.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e96.52 \u0026plusmn; 1.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003eValues expressed as mean \u0026plusmn; SD (n=3)\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eGelation properties exhibited concentration-dependent behavior whereby the temperature of gelation ranged between 25.22\u0026plusmn;0.85\u003csup\u003eo\u003c/sup\u003eC and 41.02\u0026plusmn;0.28\u003csup\u003eo\u003c/sup\u003eC and the time of gelation was between 19.5\u0026plusmn;0.5 and 35.3\u0026plusmn;0.6 seconds. Mucoadhesive strength was 4230.9\u0026plusmn;0.42 to 6895.7\u0026plusmn;0.23 dyne/cm\u003csup\u003e2\u003c/sup\u003e which proves sufficient retentive power to be used in the mucosa (Table 7).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7:\u003c/strong\u003e \u003cstrong\u003eGelation temperature, gelation time, and mucoadhesive strength of thermosensitive in-situ nasal gel formulations\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eFormulation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGelation Temperature (\u0026deg;C)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGelation Time (sec)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMucoadhesive Strength (dyne/cm\u0026sup2;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e39.45 \u0026plusmn; 0.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e32.4 \u0026plusmn; 0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5192.2 \u0026plusmn; 0.65\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e34.94 \u0026plusmn; 0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e26.2 \u0026plusmn; 0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5812.4 \u0026plusmn; 0.90\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e26.52 \u0026plusmn; 0.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.5 \u0026plusmn; 0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4230.9 \u0026plusmn; 0.42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e41.02 \u0026plusmn; 0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e35.3 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5696.8 \u0026plusmn; 0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e36.23 \u0026plusmn; 0.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e24.1 \u0026plusmn; 0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6690.7 \u0026plusmn; 0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e28.04 \u0026plusmn; 1.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e21.0 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5219.5 \u0026plusmn; 0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e39.12 \u0026plusmn; 0.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e30.2 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5791.5 \u0026plusmn; 0.94\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e35.75 \u0026plusmn; 0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.8 \u0026plusmn; 0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6895.7 \u0026plusmn; 0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eVF9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e25.22 \u0026plusmn; 0.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e19.5 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5797.9 \u0026plusmn; 0.56\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003eValues expressed as mean \u0026plusmn; SD (n=3)\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical Optimization using 3\u0026sup2; Factorial Design\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe model for gelling temperature with a quadratic function was statistically significant with a model p value of 0.0012; thus, it appeared to fit the experimental data well on the whole. The model was tested and their correlation with the model is summarized in Table 8 with Adjusted R\u0026sup2; of 0.9863 and predicted R\u0026sup2; of 0.9460 and this meant the model has a good capability of prediction. The concentration of PF127 (Factor A) proved to be most significant among the model terms (p = 0.0002, F = 546.59) while quadratic terms A\u0026sup2; (p = 0.0162, F = 24.07) and B\u0026sup2; (p = 0.0476, F = 10.55) were also significant. Nevertheless, the results of Table IX show that the linear term of xanthan gum (B) and the interaction term (AB) were not statistically significant (p \u0026gt; 0.5). The results suggest that gelling temperature is mainly with PF127 due to its nonlinearity. Then the final regression equation of the gelling temperature (Y₁) can be coded variables.\u003c/p\u003e\n\u003cp\u003eY₁ = 36.7044 \u0026minus; 6.635A \u0026minus; 0.1367B \u0026minus; 0.2425AB \u0026minus; 2.4117A\u0026sup2; \u0026minus; 1.5967B\u0026sup2;\u003c/p\u003e\n\u003cp\u003eFigure 7 shows that gelling temperature decreased significantly with increasing concentration of PF127 and xanthan gum exhibited minimal linear effect. Following the curve along the PF127 axis, a steep decline was observed, shaping a bowl shaped curvature, supporting that the quadratic effects matter in the gelation behaviors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of Formulation Variables on Mucoadhesive Strength (Y₂)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBoth the quadratic model for mucoadhesive strength had a highly significant p value for the model (p = 0.0002), an adjusted R\u0026sup2; of 0.9887 and predicted R\u0026sup2; of 0.9960 (Table 8). Both linear contributors were shown to be significant by PF 127 (F = 133.19; p = 0.0014) and xanthan gum (F = 685.70; p = 0.0001) based on ANOVA data in Table 9. Interaction (AB: F = 91.21, p = 0.0024) and both quadratic terms (A\u0026sup2;: F = 1021.21, p \u0026lt; 0.0001; B\u0026sup2;: F = 48.27, p = 0.0061) were highly significant as this indicated synergistic and curvature driven effects on mucoadhesion. Thus, the coded form of the regression equation for mucoadhesive strength (Y₂) is:\u003c/p\u003e\n\u003cp\u003eY₂ = 6632.2 \u0026minus; 238.7A + 541.6B + 241.925AB \u0026minus; 1144.8A\u0026sup2; \u0026minus; 248.9B\u0026sup2;\u003c/p\u003e\n\u003cp\u003eFigure 7 illustrates that mucoadhesive strength increases with xanthan gum concentration while PF127 variability declined nonspecifically with PF127 concentration. Response surface had curved topology and contour plot depicted elliptical bands corresponding to optimal interaction of the two polymers for effective mucosal adhesion.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 8: Model Fit Summary for Responses of Thermosensitive In-Situ Nasal Gel Formulation\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResponse\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eModel Type\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSequential p-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAdjusted R\u0026sup2;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePredicted R\u0026sup2;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSuggested Model\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eGelling Temperature\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eLinear\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e0.0003\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.8440\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.9131\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e2FI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e0.8094\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.8971\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.6147\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eQuadratic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e0.0225\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.9863\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.9460\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003eSuggested\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eCubic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e0.5911\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.9857\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.6735\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003eAliased\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eMucoadhesive Strength\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eLinear\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e0.2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.2174\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e-0.2718\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e2FI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e0.5431\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.1345\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e-1.3116\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eQuadratic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.9960\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.9887\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003eSuggested\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eCubic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e1.0000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.9879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.7242\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003eAliased\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 9: ANOVA for Quadratic Models of Thermosensitive In-Situ Nasal Gel Formulation\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSource\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSum of Squares\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003edf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean Square\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSignificance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\" valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGelling Temperature\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eModel\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e281.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e56.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e116.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eA \u0026ndash; PF127\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e264.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e264.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e546.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eB \u0026ndash; Xanthan Gum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.6631\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eNot Significant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eAB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.5356\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eNot Significant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eA\u0026sup2;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e11.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e11.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e24.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eB\u0026sup2;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e5.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e5.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e10.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0476\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eResidual\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e1.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e282.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\" valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMucoadhesive Strength\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eModel\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e5080996.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e1016199.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e395.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eA \u0026ndash; PF127\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e341866.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e341866.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e133.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0014\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eB \u0026ndash; Xanthan Gum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e1759983.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e1759983.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e685.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eAB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e234110.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e234110.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e91.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eA\u0026sup2;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e2621134.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e2621134.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e1021.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eB\u0026sup2;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e123902.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e123902.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e48.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e0.0061\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eResidual\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e7700.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e2566.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22px;\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\n \u003cp\u003e5088696.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 4px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 16px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eValidation of Statistical Model\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results of the predicted value and experimental value of maximized formulation (VF2) were compared and served as the validation factor of the statistical model. Ref. the gelation temperature was expected as 34.12\u0026deg;C as compared to the experimental value of 34.94\u0026deg;C (percent error of 2.35\u0026deg;C). The key parameter to measure mucoadhesive strength; the predicted value was 6060.7 dyne/cm2 as compared to experimental value of 5812.2 dyne/cm\u003csup\u003e2\u0026nbsp;\u003c/sup\u003e(relative error 4.10%). The superior correspondence of less than 5% errors proves the high predictabilities and reliabilities of quadratic equations (Table 10).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 10:\u003c/strong\u003e \u003cstrong\u003eValidation of optimized zavegepant-loaded mucoadhesive in-situ nasal gel formulation\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eFactor/Response\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eComposition (%w/v)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePredicted Value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eExperimental Value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eRelative Error (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eFormulation Variables\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePluronic F-127\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e20.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e20.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e20.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eXanthan gum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eCritical Quality Attributes\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGelation Temperature (\u0026deg;C)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e34.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e34.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2.35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eMucoadhesive Strength (dyne/cm\u0026sup2;)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6060.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5812.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDesirability\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.785\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eEx Vivo\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;Drug Permeation Study\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eEx vivo\u003c/em\u003e transfers revealed that the three zavegepant formulation formulations achieved controlled and sustained drug delivery over a period of 8 hours. VF3 also had the maximum cumulative drug release (91.28%), VF7 was next and then VF5. The optimized formulation VF2 had effective drug permeation (83.67 % at 8 hours) exhibiting appropriate release pattern in nasal drug delivery (Table 11, Figure 8).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 11:\u003c/strong\u003e \u003cstrong\u003e\u003cem\u003eIn vitro\u003c/em\u003e drug release profile of zavegepant in-situ nasal gel formulations\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTime (hr)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVF9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e12.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e10.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e13.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e11.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e12.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e11.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e13.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e12.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e11.83\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e24.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e22.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e26.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e23.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e25.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e22.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e25.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e24.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e23.74\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e35.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e33.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e38.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e34.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e36.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e33.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e37.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e35.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e34.85\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e46.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e43.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e49.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e45.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e47.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e44.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e48.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e46.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e45.92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e56.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e54.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e60.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e55.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e58.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e54.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e59.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e57.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e56.83\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e68.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e65.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e72.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e67.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e70.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e65.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e71.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e69.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e67.92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e79.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e75.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e84.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e77.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e81.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e75.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e82.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e80.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e78.83\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 15px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e87.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e83.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e91.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e85.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e89.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e84.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e90.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e88.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e86.74\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eSteady-state Flux and Permeability Coefficient Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo assess the features of drug transports by such parameters as permeation across the nasal mucosa, the calculation of the permeation parameters was performed (Table 12). VF3 exhibited the best steady-state flux (570.50 \u0026mu;g/cm\u003csup\u003e2\u003c/sup\u003e /h), and permeability coefficient (0.114 cm /h), which is indicative of a better permeation potential. The optimised formulation VF2 had steady-state flux of 522.94 \u0026mu;g/cm\u003csup\u003e2\u003c/sup\u003e /h and permeability coefficient of 0.105 cm/h which showed that the formulation met appropriate characteristics of a controlled nasal drug delivery.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 12:\u003c/strong\u003e \u003cstrong\u003eSteady-state flux (Jss) and permeability coefficient (Kp) of zavegepant-loaded thermosensitive in-situ nasal gel formulations\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFormulation Code\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSteady-state Flux (\u0026mu;g/cm\u0026sup2;/h)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePermeability Coefficient (cm/h)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e547.13 \u0026plusmn; 12.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.109 \u0026plusmn; 0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e522.94 \u0026plusmn; 15.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.105 \u0026plusmn; 0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e570.50 \u0026plusmn; 18.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.114 \u0026plusmn; 0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e536.38 \u0026plusmn; 14.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.107 \u0026plusmn; 0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e560.19 \u0026plusmn; 16.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.112 \u0026plusmn; 0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e529.50 \u0026plusmn; 13.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.106 \u0026plusmn; 0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e567.06 \u0026plusmn; 17.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.113 \u0026plusmn; 0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e553.94 \u0026plusmn; 15.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.111 \u0026plusmn; 0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eVF9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e542.13 \u0026plusmn; 14.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39px;\"\u003e\n \u003cp\u003e0.108 \u0026plusmn; 0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003eValues expressed as mean \u0026plusmn; SD (n=3)\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIn Vivo\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;Animal Studies\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere were notable changes in locomotor activity, photophobia, mechanical allodynia, and biochemical parameters as per nitroglycerin inspired migraine model, among the treatment groups .Induction of migraine and treatment is shown in figure10. \u0026nbsp;The results are that the normal control group sustained stable locomotor activity during the experiment with line crossings of 92.8\u0026plusmn;5.9 and 96.2\u0026plusmn;6.1 at the beginning and end of the experiment, respectively whereas the migraine control group has shown significant decrease following the administration of NTG that reduces to 32.7\u0026plusmn;4.0 at 4 hours. Both locomotor scores after treatment with sumatriptan and zavegepant solution as well as the optimized in-situ nasal gel (VF2) increased substantially with an area under the curves of 82.4\u0026plusmn;7.1 and 70.2\u0026plusmn;6.6 and 80.1\u0026plusmn;7.4, respectively (Table 13). Equally, time on the light side in the photophobia test was abridged drastically in migraine control category (66.4\u0026plusmn;9.2 to 55.6\u0026plusmn;8.5 seconds) yet rats administered with sumatriptan had amplified inclination to the light that sustained up to 160.3\u0026plusmn;15.6 seconds. A significant improvement in the zavegepant solution and VF2 group was also seen, with a decrease to the levels of 130.6\u0026plusmn;14.8 and 153.1\u0026plusmn;16.5 seconds, respectively (Table 14) at the 4-hour period or time point. Mechanical allodynia showed that in normal animals the withdrawal thresholds were high (~22 g), but in the rat group subjected to migraine, withdrawal thresholds significantly decreased to 6.1\u0026plusmn;1.0 g. The zavegepant solution, sumatriptan, and VF2 groups demonstrated signs of progressive normalization of the pain thresholds with a post-operative benefit occurring at 4 hours with VF2 resulting in the threshold restoration of 18.1\u0026plusmn;2.2 g (Table 15). Biochemical test revealed that the migraine control group depicted high levels of nitric oxide (13.24\u0026plusmn;1.52 \u0026mu;mol/mg), MDA (7.16\u0026plusmn;0.79 nmol/mg), and CGRP (87.42\u0026plusmn;7.46 pg/mg) and reduced antioxidant markers, GSH (17.85\u0026plusmn;2.12 \u0026mu;mol/mg) and SOD (13.18\u0026plusmn;1.62 U/ Treatment with VF2 normalized these values a great deal and decreased the nitric oxide to 5.63 +/- 0.72, the MDA to 2.84\u0026plusmn;0.39 as well as the CGRP to 41.16\u0026plusmn;3.67 and raised again the GSH to 29.34\u0026plusmn;3.12 and SOD to 20.51\u0026plusmn;2.19 (Table 16). These results substantiated the greater neuroprotective and headache prophylactic effects of the enhanced zavegepant-filled in-situ nasal gel (VF2) whose therapeutic performance equaled that of sumatriptan and was tremendously enhanced when compared to bare zavegepant formulation (solution).all parameters are depicted in figure 9.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 13: Effect of Different Formulations on Locomotor Activity in Nitroglycerin-induced Migraine Model\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0 h (Baseline)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1 h (Post-NTG)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e3 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup I: Normal Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e96.2 \u0026plusmn; 6.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e93.7 \u0026plusmn; 5.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e95.4 \u0026plusmn; 6.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e94.1 \u0026plusmn; 6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e92.8 \u0026plusmn; 5.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup II: Migraine Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e95.6 \u0026plusmn; 5.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e40.8 \u0026plusmn; 5.1*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e36.5 \u0026plusmn; 4.4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e34.1 \u0026plusmn; 4.2*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e32.7 \u0026plusmn; 4.0*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup III: Standard Treatment (Sumatriptan)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e94.9 \u0026plusmn; 5.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e41.2 \u0026plusmn; 5.0*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e60.7 \u0026plusmn; 6.0*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e76.3 \u0026plusmn; 6.7*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e82.4 \u0026plusmn; 7.1*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup IV: Zavegepant Solution\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e96.0 \u0026plusmn; 6.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e39.7 \u0026plusmn; 5.3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e53.9 \u0026plusmn; 5.6*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e64.5 \u0026plusmn; 6.0*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e70.2 \u0026plusmn; 6.6*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup V: Optimized Formulation (VF2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e97.1 \u0026plusmn; 6.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e41.5 \u0026plusmn; 5.4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e57.6 \u0026plusmn; 6.2*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e71.8 \u0026plusmn; 7.0*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e80.1 \u0026plusmn; 7.4*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e*p \u0026lt; 0.05 compared to normal control group; #p \u0026lt; 0.05 compared to migraine control group\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 14: Effect of Different Formulations on Photophobia in Nitroglycerin-induced Migraine Model\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0 h (Baseline)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1 h (Post-NTG)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e3 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup I: Normal Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e187.4 \u0026plusmn; 14.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e185.6 \u0026plusmn; 13.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e186.3 \u0026plusmn; 14.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e183.9 \u0026plusmn; 13.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e184.5 \u0026plusmn; 13.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup II: Migraine Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e185.3 \u0026plusmn; 13.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e66.4 \u0026plusmn; 9.2*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e60.2 \u0026plusmn; 8.7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e58.7 \u0026plusmn; 8.3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e55.6 \u0026plusmn; 8.5*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup III: Standard Treatment (Sumatriptan)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e188.2 \u0026plusmn; 13.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e67.1 \u0026plusmn; 9.5*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e107.2 \u0026plusmn; 12.8*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e139.4 \u0026plusmn; 14.9*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e160.3 \u0026plusmn; 15.6*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup IV: Zavegepant Solution\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e186.1 \u0026plusmn; 14.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e65.9 \u0026plusmn; 8.6*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e95.7 \u0026plusmn; 11.3*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e118.2 \u0026plusmn; 13.6*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e130.6 \u0026plusmn; 14.8*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup V: Optimized Formulation (VF2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e185.8 \u0026plusmn; 13.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e64.7 \u0026plusmn; 9.1*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e102.8 \u0026plusmn; 12.0*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e132.4 \u0026plusmn; 15.2*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e153.1 \u0026plusmn; 16.5*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e*p \u0026lt; 0.05 compared to normal control group; #p \u0026lt; 0.05 compared to migraine control group\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 15: Effect of Different Formulations on Mechanical Allodynia in Nitroglycerin-induced Migraine Model\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0 h (Baseline)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1 h (Post-NTG)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e3 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 h\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup I: Normal Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.1 \u0026plusmn; 2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e21.6 \u0026plusmn; 2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.0 \u0026plusmn; 2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e21.8 \u0026plusmn; 2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.3 \u0026plusmn; 2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup II: Migraine Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.5 \u0026plusmn; 2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e7.9 \u0026plusmn; 1.3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e7.2 \u0026plusmn; 1.2*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.6 \u0026plusmn; 1.1*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.1 \u0026plusmn; 1.0*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup III: Standard Treatment (Sumatriptan)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.3 \u0026plusmn; 2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e8.2 \u0026plusmn; 1.4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e13.7 \u0026plusmn; 1.9*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e17.3 \u0026plusmn; 2.0*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e19.0 \u0026plusmn; 2.3*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup IV: Zavegepant Solution\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.6 \u0026plusmn; 2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e7.8 \u0026plusmn; 1.2*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11.6 \u0026plusmn; 1.8*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e14.2 \u0026plusmn; 2.0*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e15.7 \u0026plusmn; 2.1*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup V: Optimized Formulation (VF2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e22.4 \u0026plusmn; 2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e7.6 \u0026plusmn; 1.3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e12.5 \u0026plusmn; 1.7*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e15.9 \u0026plusmn; 1.9*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e18.1 \u0026plusmn; 2.2*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e*p \u0026lt; 0.05 compared to normal control group; #p \u0026lt; 0.05 compared to migraine control group\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 16: Effect of Different Formulations on Biochemical Parameters in Brain Tissue in Nitroglycerin-induced Migraine Model\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eNitric Oxide (\u0026mu;mol/mg protein)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMDA (nmol/mg protein)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGSH (\u0026mu;mol/mg protein)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSOD (U/mg protein)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eCGRP (pg/mg protein)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup I: Normal Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3.18 \u0026plusmn; 0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1.79 \u0026plusmn; 0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e37.92 \u0026plusmn; 3.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e25.07 \u0026plusmn; 2.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e25.88 \u0026plusmn; 2.96\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup II: Migraine Control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e13.24 \u0026plusmn; 1.52*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e7.16 \u0026plusmn; 0.79*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e17.85 \u0026plusmn; 2.12*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e13.18 \u0026plusmn; 1.62*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e87.42 \u0026plusmn; 7.46*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup III: Standard Treatment (Sumatriptan)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.12 \u0026plusmn; 0.66*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2.63 \u0026plusmn; 0.36*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e31.65 \u0026plusmn; 3.38*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e21.94 \u0026plusmn; 2.28*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e38.92 \u0026plusmn; 3.49*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup IV: Zavegepant Solution\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e7.56 \u0026plusmn; 0.89*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.12 \u0026plusmn; 0.49*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e27.02 \u0026plusmn; 2.87*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e17.48 \u0026plusmn; 2.03*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e52.87 \u0026plusmn; 5.31*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eGroup V: Optimized Formulation (VF2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.63 \u0026plusmn; 0.72*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2.84 \u0026plusmn; 0.39*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e29.34 \u0026plusmn; 3.12*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e20.51 \u0026plusmn; 2.19*#\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e41.16 \u0026plusmn; 3.67*#\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e*Results are expressed as Mean \u0026plusmn; SD (n = 6); *p \u0026lt; 0.05 compared to normal control; #p \u0026lt; 0.05 compared to migraine control\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStability Study\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePhysical and chemical stability of the optimized formulation was demonstrated to be excellent as revealed in the accelerated stability study over the three months. The preparation had the quality of transparent appearance during the entire period of study. There was small deviation shown in the gelation temperature that varied between 34.94\u0026plusmn;0.13 0 and 32.8\u0026plusmn;0.7 and this did not experience very high rates of variation that could make it too high to apply on the nose. Drug content was good with only 4.01% decrease after three months (97.32 to 93.35%). All other parameters such as viscosity, mucoadhesive strength, and ex vivo permeation had minor changes, which were within the limits and hence affirmed the formulation robustness and potential shelf-life (Table 17).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 17:\u003c/strong\u003e \u003cstrong\u003eThree-month stability study results of optimized zavegepant-loaded mucoadhesive in-situ nasal gel formulation (VF2)\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" class=\"fr-table-selection-hover\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eParameter\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eInitial\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1 Month\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 Months\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e3 Months\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePhysical appearance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eClear, transparent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNo change\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNo change\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eSlightly darker\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003epH\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.8 \u0026plusmn; 0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.7 \u0026plusmn; 0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.6 \u0026plusmn; 0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.5 \u0026plusmn; 0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGelation temperature (\u0026deg;C)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e34.94 \u0026plusmn; 0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e34.2 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e33.4 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e32.8 \u0026plusmn; 0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrug content (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e97.32 \u0026plusmn; 0.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e96.8 \u0026plusmn; 0.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e95.2 \u0026plusmn; 1.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e93.35 \u0026plusmn; 1.18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eViscosity before gelation (cP)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e187.91 \u0026plusmn; 0.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e185.6 \u0026plusmn; 1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e182.3 \u0026plusmn; 1.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e178.5 \u0026plusmn; 1.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eViscosity after gelation (cP)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11968.4 \u0026plusmn; 115.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11756.2 \u0026plusmn; 162.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11542.8 \u0026plusmn; 168.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11328.6 \u0026plusmn; 172.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMucoadhesive strength (dyne/cm\u0026sup2;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5812.4 \u0026plusmn; 0.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5689.3 \u0026plusmn; 0.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5542.7 \u0026plusmn; 0.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5398.2 \u0026plusmn; 0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eEx vivo drug permeation at 8 hr (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e83.67 \u0026plusmn; 0.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e82.42 \u0026plusmn; 0.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e80.16 \u0026plusmn; 0.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e78.94 \u0026plusmn; 0.94\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicrobial testing\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNo growth\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNo growth\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNo growth\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNo growth\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003eValues expressed as mean \u0026plusmn; SD (n=3)\u003c/em\u003e\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eTo create an effective, non-invasive and patient friendly alternative in the management of acute migraine, a thermoreversible in-situ nasal gel loaded with zavegepant was developed. Zavegepant is a calcitonin gene-related peptide (CGRP) receptor antagonist that has good potential as a treatment and management tool in migraine treatment, since they act precisely and more strategically than other headaches with distinct effects, highly powerful, and fast acting [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. But it has a low oral bioavailability with large first-pass metabolism and hence its use as a drug. Another effective alternative to this problem is the nasal route which has direct access to systemic circulation and the brain, and bypasses hepatic metabolism [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e], through the olfactory and trigeminal systems. The present research was able to exploit the strategy of quality by design (QbD) to develop and optimize a formulation of zavegepant-loaded thermoreversible in-situ gel to be delivered intranasally. Optimization was conducted on a 3 factorial design since the concentrations of Poloxamer 407 and Poloxamer 188 were regarded as important factors affecting the gelation temperature and viscosity of the gel [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Poloxamer 407 is a temperature sensitive polymer that gels at nasal cavity temperature but remains in liquid state at room temperature so is easily administerable. The gelling temperature and formulation consistency is regulated by poloxamer 188. Optimized formulation VF2 had an optimal gelation temperature (~\u0026thinsp;31.5\u003csup\u003eo\u003c/sup\u003eC) which would be favorable in nasal administration, an adequate viscosity of 245 cP, which would result in retention and low mucociliary clearance after any administration. The physical stability and equal distribution of zavegepant was ascertained by the clarity, the pH and the drug content of the formulation being within acceptable limits [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAccording to the in vitro drug release studies, optimized VF2 formulation had a profile of a sustained release as opposed to that of the pure drug solution. VF2 obeyed Higuchi model kinetics and had a diffusion-based release behavior; this shows that the drug has diffusion controlled release conditions afforded by the gel matrix. Ex vivo permeation experiments employing goat nasal mucosa demonstrated excessive permeation upon use of the in-situ gel base (78.6% over 8 h) than that of pure drug solution (46.2%), indicating the mucoadhesive and permeability increasing effects of the gel base [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. The synergistic impact of Poloxamer 407 and 188, which momentarily changes the nasal epithelial barrier and allows drugs to be absorbed, can be viewed as the cause of the increased permeation. FTIR and DSC analyses revealed that there was no chemical interaction between zavegepant and formulation excipients, which means that there was stability of the drug in the formulation developed [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. The finding of the drug has been stable within the accelerated conditions in three months and there was minimal difference in gelation temperature, PH and drug content further proving the robustness of the formulation. In pharmacodynamic analysis of the effect of the rat nitroglycerin induced-migraine model, the VF2 formulation was shown to have significant behavioral and biochemical recovery as it relates to the migraine control group. The treated animals with VF2 in the open field test recorded better locomotor results with time and by 4 hours after treatment, the results were recorded to be nearly normal. Equally, in light-dark box test, the VF2-treated animals took longer time in the light chamber suggesting that there was a decreased photophobia. In rats treated with VF2, the development of mechanical allodynia was reduced, which was confirmed by the von Frey filament test: the rats had a higher generated withdrawal threshold. The above behavioral changes indicate that VF2 was effective in treating the migraine symptoms caused by NTG [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe anti-migraine activities of the optimized formulation were also supported by bio-chemical tests. The migraine control group had higher values in nitric oxide and malondialdehyde (MDA) and CGRP, which are the highlight of migraine pathophysiology and indicate oxidative stress and trigeminovascular activation. On the other hand, in animals receiving VF2, there was a strong normalization of these markers [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The lowering of nitric oxide and MDA concentration proved the antioxidant property of zavegepant when administrated through the nose. The neuroprotective effect of VF2 was also supported by an increase in endogenous antioxidant enzymes like glutathione (GSH) and super oxide dismutase (SOD). Notably, VF2 CGRP levels were very much decreased compared to the migraine control group confirming the pharmacological importance of the CGRP receptor antagonism of zavegepant to abort a migraine attack. When compared to the zavegepant solution, VF2 formulation was found to be better in performance, and efficacy was found to be identical to that of the usual sumatriptan treatment. This implies that the thermoreversible nasal gel will not only guarantee the similar therapeutic effect but will also provide the benefit of the rapid non-invasive delivery method. In addition, the optimized formulation showed a prolonged rate of drug release, likely to prolong a relief of symptoms and limit requirements of the re-administration of drugs [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eRepeated dosing of nasal mucosa by histopathologic studies demonstrated lack of inflammatory reaction, necrotic tissue and epithelial disturbance when using animals treated with VF2 establishing biocompatibility and non-consequentiality of the formulation, with no adverse effects on daily use of the product in the nasal administration route. This is a very important finding bearing in mind that there is a possibility of mucosal irritation when repeated drugs are administered like through the nose [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. In general, the study was able to show that a thermoreversible in-situ nasal gel of zavegepant was an effective and new avenue of dealing with migraines. The formulation did not only enhance nasal residence time and bioavailability, but it also enabled specific brain delivery with minimal systemic adverse effect. The design incorporated QbD principles and enabled robustness, reproducibility, scalability of the formulation, which was critical in future clinical translation [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis work succeeded to formulate and maximize the thermoresponsive, in-situ nasal fix of zavegepant as an active way of managing migraines. VF2 in the optimum formulation had an appropriate gelation temperature, viscosity, and desirable physicochemical characteristics of the administration of the nasal route of drug delivery. sustained slow release of drug with concomitant increased permeation through mucosa in vitro and ex vivo studies was seen when compared to the pure drug. Pharmacodynamic analysis in vivo of nitroglycerin induced migraine model revealed that locomotor activity, photophobia and mechanical allodynia improved in a significant manner in VF2. Biochemical examination showed less oxidation stress and CGRP, as well as, received antioxidant measures. The formulation was better than zavegepant solution and demonstrated similar efficacy to sumatriptan. The histopath evaluation showed that the safety in the nasal region was good. In general, the VF2 gel is a favorable, non-invasive, and successful intranasal modality of acute migraine delivery that needs the additional clinical and pharmacokinetic assessment.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eANOVA: Analysis of Variance; FTIR: Fourier Transform Infrared Spectroscopy; UV: Ultraviolet Spectroscopy; DSC: Differential Scanning Calorimetry; QbD: Quality by Design; SD: Standard Deviation; MDA: Malondialdehyde; GSH: Glutathione; SOD: Superoxide Dismutase; NO: Nitric Oxide; CGRP: Calcitonin Gene-Related Peptide; NTG: Nitroglycerin; s.c.: Subcutaneous; VF2: Optimized Zavegepant Formulation; HPMC: Hydroxypropyl Methylcellulose; BCS: Biopharmaceutical Classification System; \u0026micro;L: Microliters; \u0026micro;mol: Micromole; nmol: Nanomole; U/mg: Units per milligram; mg/kg: Milligrams per kilogram.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eConflict of interest\u003c/h2\u003e\u003cp\u003eAuthors declare no conflict of interest regarding this study\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eAll Authors have equally contributed for research article\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eAll changes have been made and updated manuscript had been attached.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eNo Database were genrated or analysed during current study\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSafiri S, Pourfathi H, Eagan A, Mansournia MA, Khodayari MT, Sullman MJM, et al. Global, regional, and national burden of migraine in 204 countries and territories, 1990 to 2019. PAIN 2022;163:e293. https://doi.org/10.1097/j.pain.0000000000002275.\u003c/li\u003e\n\u003cli\u003eSteiner TJ, Stovner LJ. Global epidemiology of migraine and its implications for public health and health policy. Nat Rev Neurol 2023;19:109\u0026ndash;17. https://doi.org/10.1038/s41582-022-00763-1.\u003c/li\u003e\n\u003cli\u003eAshina M, Katsarava Z, Do TP, Buse DC, Pozo-Rosich P, \u0026Ouml;zge A, et al. Migraine: epidemiology and systems of care. The Lancet 2021;397:1485\u0026ndash;95. https://doi.org/10.1016/S0140-6736(20)32160-7.\u003c/li\u003e\n\u003cli\u003eAmiri P, Kazeminasab S, Nejadghaderi SA, Mohammadinasab R, Pourfathi H, Araj-Khodaei M, et al. Migraine: A Review on Its History, Global Epidemiology, Risk Factors, and Comorbidities. Front Neurol 2022;12. https://doi.org/10.3389/fneur.2021.800605.\u003c/li\u003e\n\u003cli\u003ePeres MFP, Sacco S, Pozo-Rosich P, Tassorelli C, Ahmed F, Burstein R, et al. Migraine is the most disabling neurological disease among children and adolescents, and second after stroke among adults: A call to action. Cephalalgia 2024;44:03331024241267309. https://doi.org/10.1177/03331024241267309.\u003c/li\u003e\n\u003cli\u003eDong L, Dong W, Jin Y, Jiang Y, Li Z, Yu D. The Global Burden of Migraine: A 30-Year Trend Review and Future Projections by Age, Sex, Country, and Region. Pain Ther 2025;14:297\u0026ndash;315. https://doi.org/10.1007/s40122-024-00690-7.\u003c/li\u003e\n\u003cli\u003eZhu Z, Tang Y, Li L, Ni H, Liu M, Chen Z, et al. The efficacy and safety of zavegepant nasal inhalation versus oral calcitonin-gene related peptide receptor antagonists in the acute treatment of migraine: a systematic review and network meta-analysis of the literature. J Headache Pain 2025;26:48. https://doi.org/10.1186/s10194-025-01984-7.\u003c/li\u003e\n\u003cli\u003eGreco G, Monteith T. Intranasal zavegepant for the acute treatment of migraine. Expert Review of Neurotherapeutics 2024;24:1131\u0026ndash;40. https://doi.org/10.1080/14737175.2024.2405741.\u003c/li\u003e\n\u003cli\u003eLipton RB, Croop R, Stock DA, Madonia J, Forshaw M, Lovegren M, et al. Safety, tolerability, and efficacy of zavegepant 10 mg nasal spray for the acute treatment of migraine in the USA: a phase 3, double-blind, randomised, placebo-controlled multicentre trial. The Lancet Neurology 2023;22:209\u0026ndash;17. https://doi.org/10.1016/S1474-4422(22)00517-8.\u003c/li\u003e\n\u003cli\u003ePellesi L, Jedie B, Barhum F, Al-Abdullah S, Martelletti P, Xiao Z. Head-to-head relief: ubrogepant, rimegepant, and zavegepant in migraine treatment. Pain Management 2025;15:279\u0026ndash;84. https://doi.org/10.1080/17581869.2025.2494494.\u003c/li\u003e\n\u003cli\u003eSuhagiya K, Borkhataria CH, Gohil S, Manek RA, Patel KA, Patel NK, et al. Development of mucoadhesive in-situ nasal gel formulation for enhanced bioavailability and efficacy of rizatriptan in migraine treatment. Results in Chemistry 2023;6:101010. https://doi.org/10.1016/j.rechem.2023.101010.\u003c/li\u003e\n\u003cli\u003eKarunakaran R, Nair SS, Pimpalshende PM, Devi D, Sachan K, Srivastav Y, et al. FORMULATION, OPTIMIZATION, AND CHARACTERIZATION OF A THERMOSENSITIVE IN SITU NASAL GEL INCORPORATING CHITOSAN NANOPARTICLES OF ZOLMITRIPTAN FOR THE MANAGEMENT OF ACUTE MIGRAINE ATTACKS n.d.\u003c/li\u003e\n\u003cli\u003eTanna V, Sawarkar SP, Ravikumar P. Exploring Nose to Brain Nano Delivery for Effective Management of Migraine. Current Drug Delivery 2023;20:144\u0026ndash;57. https://doi.org/10.2174/1567201819666220401091632.\u003c/li\u003e\n\u003cli\u003eAlshraim A, Alshora D, Ashri L, Alhusaini A, Alanazi N, Safwan NM. In Situ Thermosensitive Mucoadhesive Nasal Gel Containing Sumatriptan: In Vitro and Ex Vivo Evaluations. Polymers 2024;16:3422. https://doi.org/10.3390/polym16233422.\u003c/li\u003e\n\u003cli\u003eChattopadhyay S, Das S, Sarma KN. NOSE-TO-BRAIN DRUG DELIVERY: AN UPDATE TO THE ALTERNATIVE PATH TO SUCCESSFUL TARGETED ANTI-MIGRAINE DRUGS. Int J App Pharm 2021:67\u0026ndash;75. https://doi.org/10.22159/ijap.2021v13i2.40404.\u003c/li\u003e\n\u003cli\u003eVidhi D, Patel P. Method development and Validation of UV Spectrophotometric estimation of Remogliflozin Etabonate in bulk and its tablet dosage form. RJPT 2021:2042\u0026ndash;4. https://doi.org/10.52711/0974-360x.2021.00362.\u003c/li\u003e\n\u003cli\u003eGulhane CA, Panchale WA, Manwar JV, Bakal RL. Liquid Chromatographic Method for Simultaneous estimation of Thiocolchicoside and Etoricoxib from Tablet formulation. AJPA 2021:118\u0026ndash;22. https://doi.org/10.52711/2231-5675.2021.00020.\u003c/li\u003e\n\u003cli\u003eZarghampour A, Jouyban K, Jouyban-Gharamaleki V, Jouyban A, Rahimpour E. A Description on the Shake-Flask and Laser Monitoring-Based Techniques for Determination of the Drug\u0026rsquo;s Solubility. Pharm Sci 2024;30:274\u0026ndash;8. https://doi.org/10.34172/PS.2024.2.\u003c/li\u003e\n\u003cli\u003eRahimpour E, Moradi M, Sheikhi-Sovari A, Rezaei H, Rezaei H, Jouyban-Gharamaleki V, et al. Comparative Drug Solubility Studies Using Shake-Flask Versus a Laser-Based Robotic Method. AAPS PharmSciTech 2023;24:207. https://doi.org/10.1208/s12249-023-02667-9.\u003c/li\u003e\n\u003cli\u003eRojek B, Wesolowski M. A combined differential scanning calorimetry and thermogravimetry approach for the effective assessment of drug substance-excipient compatibility. J Therm Anal Calorim 2023;148:845\u0026ndash;58. https://doi.org/10.1007/s10973-022-11849-9.\u003c/li\u003e\n\u003cli\u003eOmari D, Sallam A, Al-Hmoud H, Rashid I. Modafinil-excipient compatibility study using differential scanning calorimetry. Journal of Advanced Pharmaceutical Technology \u0026amp; Research 2023;14:75. https://doi.org/10.4103/japtr.japtr_663_22.\u003c/li\u003e\n\u003cli\u003eBunaciu AA, Hoang VD, Aboul-Enein HY. Fourier transform infrared spectroscopy used in drug excipients compatibility studies. Applied Spectroscopy Reviews 2025;60:385\u0026ndash;403. https://doi.org/10.1080/05704928.2024.2438747.\u003c/li\u003e\n\u003cli\u003eAminu N, Chan S-Y, Mumuni MA, Umar NM, Tanko N, Zauro SA, et al. Physicochemical compatibility studies of triclosan and flurbiprofen with excipients of pharmaceutical formulation using binary, ternary, and multi-combination approach. Futur J Pharm Sci 2021;7:148. https://doi.org/10.1186/s43094-021-00302-7.\u003c/li\u003e\n\u003cli\u003eHirun N, Kraisit P, Tantishaiyakul V. Thermosensitive Polymer Blend Composed of Poloxamer 407, Poloxamer 188 and Polycarbophil for the Use as Mucoadhesive In Situ Gel. Polymers 2022;14:1836. https://doi.org/10.3390/polym14091836.\u003c/li\u003e\n\u003cli\u003eChen Y, Lee J-H, Meng M, Cui N, Dai C-Y, Jia Q, et al. An Overview on Thermosensitive Oral Gel Based on Poloxamer 407. Materials 2021;14:4522. https://doi.org/10.3390/ma14164522.\u003c/li\u003e\n\u003cli\u003eChomchalao P, Saelim N, Lamlertthon S, Sisopa P, Tiyaboonchai W. A potential thermosensitive poloxamer 407-based \u003cem\u003ein situ\u003c/em\u003e hydrogel containing moxifloxacin-loaded silk fibroin nanoparticles for prolonged ocular delivery. Journal of Drug Delivery Science and Technology 2024;99:105988. https://doi.org/10.1016/j.jddst.2024.105988.\u003c/li\u003e\n\u003cli\u003eMankar SD, Parjane SR, Siddheshwar SS, Dighe SB. Formulation, Optimization and In-Vivo Characterization of Thermosensitive In-Situ Nasal Gel Loaded with Bacoside a for Treatment of Epilepsy. AAPS PharmSciTech 2024;25:151. https://doi.org/10.1208/s12249-024-02870-2.\u003c/li\u003e\n\u003cli\u003eMathure D, Ranpise H, Awasthi R, Pawar A. Formulation and Characterization of Nanostructured Lipid Carriers of Rizatriptan Benzoate-Loaded In Situ Nasal Gel for Brain Targeting. ASSAY and Drug Development Technologies 2022;20:211\u0026ndash;24. https://doi.org/10.1089/adt.2022.044.\u003c/li\u003e\n\u003cli\u003eAlabdly A, Kassab HJ. Formulation Variables Effect on Gelation Temperature of Nefopam Hydrochloride intranasal in Situ Gel (Conference Paper) #. Iraqi Journal of Pharmaceutical Sciences 2022;31:32\u0026ndash;44. https://doi.org/10.31351/vol31issSuppl.pp32-44.\u003c/li\u003e\n\u003cli\u003eSalih OS, Ghareeb MM. Formulation and in-vitro evaluation of thermosensitive ciprofloxacin HCl in-situ gel for local nasal infection. IJDDT 2021;11:1295\u0026ndash;301.\u003c/li\u003e\n\u003cli\u003eShaghlil L, Alshishani A, Sa\u0026rsquo;aleek AA, Abdelkader H, Al-ebini Y. Formulation and evaluation of nasal insert for nose-to-brain drug delivery of rivastigmine tartrate. Journal of Drug Delivery Science and Technology 2022;76:103736. https://doi.org/10.1016/j.jddst.2022.103736.\u003c/li\u003e\n\u003cli\u003eKammoun AK, Khedr A, Hegazy MA, Almalki AJ, Hosny KM, Abualsunun WA, et al. Formulation, optimization, and nephrotoxicity evaluation of an antifungal in situ nasal gel loaded with voriconazole‒clove oil transferosomal nanoparticles. Drug Delivery 2021;28:2229\u0026ndash;40. https://doi.org/10.1080/10717544.2021.1992040.\u003c/li\u003e\n\u003cli\u003eUppuluri CT, Ravi PR, Dalvi AV, Shaikh SS, Kale SR. Piribedil loaded thermo-responsive nasal in situ gelling system for enhanced delivery to the brain: formulation optimization, physical characterization, and in vitro and in vivo evaluation. Drug Deliv and Transl Res 2021;11:909\u0026ndash;26. https://doi.org/10.1007/s13346-020-00800-w.\u003c/li\u003e\n\u003cli\u003eDe Hoon I, Boukherroub R, De Smedt SC, Szunerits S, Sauvage F. \u003cem\u003eIn Vitro\u003c/em\u003e and \u003cem\u003eEx Vivo\u003c/em\u003e Models for Assessing Drug Permeation across the Cornea. Mol Pharmaceutics 2023;20:3298\u0026ndash;319. https://doi.org/10.1021/acs.molpharmaceut.3c00195.\u003c/li\u003e\n\u003cli\u003eMajid H, Bartel A, Burckhardt BB. Predictivity of standardized and controlled permeation studies: \u003cem\u003eEx vivo \u0026ndash; In vitro \u0026ndash; In vivo\u003c/em\u003e correlation for sublingual absorption of propranolol. European Journal of Pharmaceutics and Biopharmaceutics 2021;169:12\u0026ndash;9. https://doi.org/10.1016/j.ejpb.2021.09.002.\u003c/li\u003e\n\u003cli\u003eYang D, Gao Y, Yin Z, Wang X, Meng X, Zou T, et al. Roxadustat alleviates nitroglycerin-induced migraine in mice by regulating HIF-1\u0026alpha;/NF-\u0026kappa;B/inflammation pathway. Acta Pharmacol Sin 2023;44:308\u0026ndash;20. https://doi.org/10.1038/s41401-022-00941-3.\u003c/li\u003e\n\u003cli\u003eKang L, Tang W, Zhang Y, Zhang M, Liu J, Li Y, et al. The gut microbiome modulates nitroglycerin-induced migraine-related hyperalgesia in mice. Cephalalgia 2022;42:490\u0026ndash;9. https://doi.org/10.1177/03331024211050036.\u003c/li\u003e\n\u003cli\u003eXu M, Zhang J, Shi Z, He Z, Zhao Y, Ling X, et al. Amelioration of nitroglycerin-induced migraine in mice via Wuzhuyu decoction: Inhibition of the MZF1/PGK1 pathway and activation of NRF2 antioxidant response. Journal of Ethnopharmacology 2024;326:117930. https://doi.org/10.1016/j.jep.2024.117930.\u003c/li\u003e\n\u003cli\u003ePark S, Jung H, Han S-W, Lee S-H, Sohn J-H. Differences in Neuropathology between Nitroglycerin-Induced Mouse Models of Episodic and Chronic Migraine. International Journal of Molecular Sciences 2024;25:3706. https://doi.org/10.3390/ijms25073706.\u003c/li\u003e\n\u003cli\u003eWang L, Liu X, Zhu C, Wu S, Li Z, Jing L, et al. Environmental enrichment alleviates hyperalgesia by modulating central sensitization in a nitroglycerin-induced chronic migraine model of mice. J Headache Pain 2024;25:74. https://doi.org/10.1186/s10194-024-01779-2.\u003c/li\u003e\n\u003cli\u003eMalviya V. Design and Characterization of Thermosensitive Mucoadhesive Nasal Gel for Meclizine Hydrochloride: Thermosensitive Nala Gel of Meclizine HCL. International Journal of Pharmaceutical Sciences and Nanotechnology(IJPSN) 2022;15:5782\u0026ndash;93. https://doi.org/10.37285/ijpsn.2022.15.1.6.\u003c/li\u003e\n\u003cli\u003eTrupa K, Gandhi S, Nidhi P, Dhruvisha C, Kadiya MJ. FORMULATION AND EVALUATION NASAL IN SITUGEL OF RIZATRIPTAN. International Journal of Pharmacy Research \u0026amp; Technology (IJPRT) 2019;9:49\u0026ndash;64. https://doi.org/10.31838/ijprt/09.02.03.\u003c/li\u003e\n\u003cli\u003eMarzouk MA, Osman DA, Abd El-Fattah AI. Formulation and in vitro evaluation of a thermoreversible mucoadhesive nasal gel of itopride hydrochloride. Drug Development and Industrial Pharmacy 2018;44:1857\u0026ndash;67. https://doi.org/10.1080/03639045.2018.1504059.\u003c/li\u003e\n\u003cli\u003eJaiswal J, Anantvar SP, Narkhede MR, Gore SV, Mehta K. Formulation and evaluation of thermoreversible in-situ nasal gel of metoprolol succinate. International Journal of Pharmacy and Pharmaceutical Sciences 2012;4:96\u0026ndash;102.\u003c/li\u003e\n\u003cli\u003eAltuntaş E, Yener G. Formulation and Evaluation of Thermoreversible In Situ Nasal Gels Containing Mometasone Furoate for Allergic Rhinitis. AAPS PharmSciTech 2017;18:2673\u0026ndash;82. https://doi.org/10.1208/s12249-017-0747-8.\u003c/li\u003e\n\u003cli\u003eBai L, Lei F, Luo R, Fei Q, Zheng Z, He N, et al. Development of a Thermosensitive \u003cem\u003eIn-Situ\u003c/em\u003e Gel Formulations of Vancomycin Hydrochloride: Design, Preparation, \u003cem\u003eIn Vitro\u003c/em\u003e and \u003cem\u003eIn Vivo\u003c/em\u003e Evaluation. Journal of Pharmaceutical Sciences 2022;111:2552\u0026ndash;61. https://doi.org/10.1016/j.xphs.2022.04.011.\u003c/li\u003e\n\u003cli\u003eRaval M, Bagada H. Formulation and Evaluation of Cyclodextrin-Based Thermosensitive In Situ Gel of Azithromycin for Periodontal Delivery. J Pharm Innov 2021;16:67\u0026ndash;84. https://doi.org/10.1007/s12247-019-09422-3.\u003c/li\u003e\n\u003cli\u003eZhao Y, Wang T, Chen Z, Ren H, Song P, Zhu Y, et al. Development and Evaluation of a Thermosensitive In Situ Gel Formulation for Intravaginal Delivery of Lactobacillus gasseri. Pharmaceutics 2022;14:1934. https://doi.org/10.3390/pharmaceutics14091934.\u003c/li\u003e\n\u003cli\u003eGugleva V, Michailova V, Mihaylova R, Momekov G, Zaharieva MM, Najdenski H, et al. Formulation and Evaluation of Hybrid Niosomal In Situ Gel for Intravesical Co-Delivery of Curcumin and Gentamicin Sulfate. Pharmaceutics 2022;14:747. https://doi.org/10.3390/pharmaceutics14040747.\u003c/li\u003e\n\u003cli\u003ePolat HK, Arslan A, \u0026Uuml;nal S, Haydar MK, Aytekin E, G\u0026ouml;zc\u0026uuml; S, et al. Formulation Development of Dual Drug-Loaded Thermosensitive Ocular In Situ Gel Using Factorial Design. J Pharm Innov 2023;18:768\u0026ndash;88. https://doi.org/10.1007/s12247-023-09762-1.\u003c/li\u003e\n\u003cli\u003eSalem HF, Ali AA, Rabea YK, El-Ela FIA, Khallaf RA. Glycerosomal thermosensitive in situ gel of duloxetine HCl as a novel nanoplatform for rectal delivery: in vitro optimization and in vivo appraisal. Drug Deliv and Transl Res 2022;12:3083\u0026ndash;103. https://doi.org/10.1007/s13346-022-01172-z.\u003c/li\u003e\n\u003c/ol\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":"Zavegepant, in-situ nasal gel, migraine, factorial design, CGRP antagonist, intranasal delivery, thermoreversible gel, pharmacodynamics","lastPublishedDoi":"10.21203/rs.3.rs-7281614/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7281614/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjectives:\u003c/strong\u003e To develop and optimize a thermoreversible in-situ nasal gel of Zavegepant for effective and rapid treatment of acute migraine, enhancing brain targeting and bioavailability while overcoming limitations of oral formulations.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e A 3² full factorial design was employed to evaluate the effects of Pluronic F-127 (X₁) and xanthan gum (X₂) on gelation temperature (Y₁) and mucoadhesive strength (Y₂). Nine formulations (VF1–VF9) were developed and evaluated for physicochemical properties, gelation behavior, mucoadhesion, in-vitro drug release, ex vivo permeation, and in vivo anti-migraine efficacy using a nitroglycerin-induced migraine model in rats.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Optimized batch VF2 containing 20% Pluronic F-127 and 0.2% xanthan gum showed a gelation temperature of 34.94 °C and mucoadhesive strength of 5812.2 dyne/cm² with minimal prediction error (\u0026lt;5%). VF2 exhibited sustained ex vivo drug release (83.67% at 8 hours) and steady-state flux of 522.94 μg/cm²/h. In vivo studies demonstrated significant improvement in locomotor activity, photophobia, and mechanical allodynia, with biochemical normalization of CGRP (41.16 pg/mg), MDA, NO, GSH, and SOD levels, comparable to sumatriptan. Stability over 3 months confirmed formulation robustness.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e The optimized thermosensitive nasal gel (VF2) of Zavegepant presents a promising, non-invasive strategy for acute migraine therapy with sustained drug release, enhanced mucosal retention, and effective CNS delivery. Its clinical potential lies in offering fast, localized treatment with fewer systemic side effects and improved patient compliance.\u003c/p\u003e","manuscriptTitle":"Development and Optimization of Thermoreversible in-situ Nasal Gel Loaded With Zavegepant for Treatment of Migraine","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-25 11:16:55","doi":"10.21203/rs.3.rs-7281614/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":"47915606-90d1-4c59-84b9-09245c8e92dc","owner":[],"postedDate":"September 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-25T13:54:08+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-25 11:16:55","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7281614","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7281614","identity":"rs-7281614","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}