In this study, we present the lyophilization process development efforts for a vaccine formulation aimed at optimizing the primary drying time (hence, the total cycle length) through comprehensive evaluation of its thermal characteristics, temperature profile, and critical quality attributes (CQAs). Differential scanning calorimetry (DSC) and freeze-drying microscopy (FDM) were used to experimentally determine the product-critical temperatures, viz., the glass transition temperature (Tg\') and the collapse temperature (Tc). Initial lyophilization studies indicated that the conventional approach of targeting product temperature (Tp) below the Tc (determined from FDM) resulted in long and sub-optimal drying times. Interestingly, aggressive drying conditions where the product temperature reached the total collapse temperature did not result in macroscopic collapse but, instead, reduced the drying time by ∼ 45 % while maintaining product quality requirements. This observation suggests the need for a more reliable measurement of the macroscopic collapse temperature for product in vials. The temperature profiles from different lyophilization runs showed a drop in product temperature following the primary drying ramp, of which the magnitude was correlated to the degree of macroscopic collapse. The batch-average product resistance, Rp, determined using the manometric temperature measurement (MTM), decreased with increasing dried layer thickness for aggressive primary drying conditions. A quantitative analysis of the product temperature and resistance profiles combined with qualitative assessment of cake appearance attributes was used to determine a more representative macro-collapse temperature, Tcm, for this vaccine product. A primary drying design space was generated using first principles modeling of heat and mass transfer to enable selection of optimum process parameters and reduce the number of exploratory lyophilization runs. Overall, the study highlights the importance of accurate determination of macroscopic collapse in vials, pursuing aggressive drying based on individual product characteristics, and leveraging experimental and modeling techniques for process optimization.

The focus of this investigation was to determine the mechanism of effective glass transition temperature (TgE) on the crystallization behavior and microstructure of drugs in crystalline solid dispersion (CSD). CSDs were prepared by rotary evaporation using ketoconazole (KET) as a model drug and the triblock copolymer poloxamer 188 as a carrier. The pharmaceutical properties of CSDs, such as crystallite size, crystallization kinetics, and dissolution behavior, were investigated to provide a foundation for studying the crystallization behavior and the microstructure of drugs in CSDs. According to classical nucleation theory, the relationship of treatment temperature-drug crystallite size-TgE of CSD was investigated. Voriconazole, a compound that is structurally similar to KET but with different physicochemical properties, was used to verify the conclusions. The dissolution behavior of KET was significantly enhanced compared to the raw drug due to smaller crystallite size. Crystallization kinetic studies revealed a two-step crystallization mechanism for KET-P188-CSD, in which P188 crystallized first and KET crystallized later. When the treatment temperature was near TgE, the drug crystallite size was smaller and more numerous, which suggests nucleation and slow growth. With the increase of temperature, the drug changed from nucleation to growth, and the number of crystallites decreased and the size of the drug increased. This result suggests it is possible to prepare CSDs with higher drug loading and smaller crystallite size by adjusting the treatment temperature and TgE, so as to maximize the drug dissolution rate. The VOR-P188-CSD maintained a relationship between treatment temperature, drug crystallite size, and TgE. The findings of our study demonstrate that TgE and the treatment temperature can be used to regulate the drug crystallite size and improve the drug solubility and dissolution rate.

Coamorphization has been proven to be an effective approach to improve bioavailability of poorly soluble active pharmaceutical ingredients (APIs) by virtue of solubilization, and also contributes to overcome limitation of physical stability associated with amorphous drug alone. In current work, a co-amorphous formulation of dipyridamole (DPM), a poor solubility drug, with p-hydroxybenzoic acid (HBA) was prepared and investigated. At a molar ratio of 1:2, DPM and HBA were melted result in the formation of a binary co-amorphous system. The DPM-HBA co-amorphous was structurally characterized by powder X-ray diffraction (PXRD), temperature modulated differential scanning calorimetry (mDSC), high performance liquid chromatography (HPLC) and solution state 1H nuclear magnetic resonance (1H NMR). The molecular mechanisms in the co-amorphous were further analysed via Fourier-transform infrared (FTIR) and Raman spectroscopies, as well as density functional theory (DFT) calculation. All the results consistently revealed the presence of hydrogen bonding interactions between -OH of DPM and -COOH on HBA. Accelerated test and glass transition kinetics showed excellent physical stability of DPM-HBA co-amorphous compared with amorphous DPM along with glass transition temperatures (Tg). The phase-solubility study indicated that complexation occurred between DPM and HBA in solution, which contributed to the solubility and dissolution enhancement of DPM in co-amorphous system. Pharmacokinetic study of co-amorphous DPM-HBA in mouse plasma revealed that the DPM exhibited 1.78-fold and 2.64-fold improvement in AUC0‑∞ value compared with crystalline and amorphous DPM, respectively. This current study revealed coamorphization is an effective approach for DPM to improve the solubility and biopharmaceutical performance.

Understanding the heterogeneous nano/microscopic structures of various organic glasses is fundamental and necessary for many applications. Recently, unusual structural phenomena have been observed experimentally in various organic glasses near their glass transition temperatures (Tg), including dibutyl phthalate (DBP). In particular, the librational motion of radical probe in the glass is progressively suppressed upon temperature increase. In this work, we report in-depth molecular dynamics studies of structural anomalies in DBP glass, that revealed insights into the general mechanism of these phenomena. In particular, we have evidenced that the two types of solvation within alkyl chains coexist, allowing only small-angle wobbling of the solute molecule (TEMPO radical), and another favouring large-angle rotations. The former solvation assumes constrained location of the solute near carboxyl groups of DBP, while the latter is coupled to the concerted movement of butyl chains. Remarkably, excellent qualitative and quantitative agreement with previous experimental results were obtained. As such, we are certain that the above-mentioned dynamic phenomena explain the intriguing structural anomalies observed in DBP and some other glasses in the vicinity of Tg.

New anti-octadecaborane(22) laser dyes have been recently introduced. However, their application in solid thin films is limited, despite being very desirable for electronics. Spectroscopic methods, photoluminescence (PL), and infrared reflection-absorption spectroscopy (IRRAS), are here used to reveal structural responses to a temperature change in thin polymer films made of π- and σ-conjugated and non-conjugated polymers and anti-octadecaborane(22) and its tetra-alkylatedderivatives. It has been observed that borane clusters are not firmly fixed within polymer matrices and that their ability for diffusion out of the polymer film is unprecedented, especially at higher temperatures. This ability is related to thermodynamic transitions of polymer macromolecular chains. PL and IRRAS spectra have revealed a clear correlation with β-transition and α-transition of polymers. The influence of structure and molecular weight of a polymer and the concentration and the substitution type of clusters on mobility of borane clusters within the polymer matrix is demonstrated. A solution is proposed that led to an improvement of the temperature stability of films by 45 °C. The well-known spectroscopic methods have proved to be powerful tools for a non-routine description of the temperature behavior of both borane clusters and polymer matrices.

OBJECTIVE: The present research aimed to characterize and deduce the structure of a novel denture base copolymer containing antimicrobial triazine comonomer by nuclear magnetic resonance (NMR) and energy-dispersive X-ray (EDX) spectroscopies. Also, it aimed to evaluate the glass transition temperature (Tg) with the addition of TATA at different concentrations.
METHODS: The trial groups G10 and G20 were thermo-polymerized with triazine comonomer, whereas the control group G0 was polymerized without the triazine. NMR and EDX spectroscopies assessed copolymerization along with deducing elemental composition in 
mass %. The surface topographies were observed through field-emission scanning electron microscopy (FESEM). The Tg of the resultant copolymer was examined by differential scanning calorimetry. Pertinent statistical tests with relevant multiple comparison tests were exercised to compare the mean Tg of the groups.
RESULTS: The configuration of a new copolymer containing triazine comonomer was manifested with additional protons and carbon atoms. Nitrogen was detected in the EDX spectroscopy of the trial groups. The Tg of the new copolymer was higher than the G0. The triazine comonomer in the copolymer at 20% concentration exhibited the highest Tg.
CONCLUSIONS: The triazine comonomer substitution produced a novel denture base copolymer with enhanced Tg.
CONCLUSIONS: The novel denture base copolymer may possess enhanced biomechanical properties due to the TATA\'s cross-linking capability. Nevertheless, the antimicrobial property of the triazine comonomer incorporated in the denture base composition might be beneficial in inhibiting the microbial colonization on the denture\'s surface.

As the human excreta, urine is often used as one of the test materials in medical research due to its composition and content directly reflecting the health status of the body. Considering that the substances in urine may show different effects on its freezing process, solidification characteristics of sessile urine droplets on a horizontal cold plate surface under natural convection were experimentally investigated by comparing with those of water droplets under same conditions. To make the conclusion analysis more reasonable, the urine of a human without any diseases, especially metabolic diseases, was treated and used. The characteristics include nucleation location, dynamic variation of droplet color, and temperatures at different heights inside the droplet, and so forth. It was found that, similar to that of a water droplet, the solidification process of a urine droplet also experiences the following four stages: supercooling, recalescence, freezing, and cooling, in chronological order. Differently, the urine droplet changes from transparent to blur white at the supercooling stage due to the precipitation of inorganic salts. For nucleation locations, 46.67% cases are at the bottom, while others are at the top and middle of urine droplets. For a 10 μL droplet on a surface of -30 °C, urine has a 0.95 s freezing duration shorter than water, and a 5.31 °C lower phase-transition temperature. Results of this study are expected to reflect the content of substances in urine and thus provide references for urinalysis of patients with metabolic diseases.

Recently, glasses, a subset of amorphous solids, have gained attention in various fields, such as polymer chemistry, optical fibers, and pharmaceuticals. One of their characteristic features, the glass transition temperature (Tg) which is absent in 100% crystalline materials, influences several material properties, such as free volume, enthalpy, viscosity, thermodynamic transitions, molecular motions, physical stability, mechanical properties, etc. In addition to Tg, there may be several other temperature-dependent transitions known as sub-Tg transitions (or β-, γ-, and δ-relaxations) which are identified by specific analytical techniques. The study of Tg and sub-Tg transitions occurring in amorphous solids has gained much attention because of its importance in understanding molecular kinetics, and it requires the combination of conventional and novel characterization techniques. In the present study, three different analytical techniques [modulated differential scanning calorimetry (mDSC), dynamic mechanical analysis (DMA), and dielectric relaxation spectroscopy (DRS)] were used to perform comprehensive qualitative/quantitative characterization of molecular relaxations, miscibility, and molecular interactions present in an amorphous polymer (PVPVA), a model drug (indomethacin, IND), and IND/PVPVA-based amorphous solid dispersions (ASDs). This is the first ever reported DMA study on PVPVA in its powder form, which avoids the contribution of solvent to the mechanical properties when a self-standing polymer film is used. A good correlation between the techniques in determining the Tg value of PVPVA, IND, and IND/PVPVA-based ASDs is established, and the negligible difference (within 10 °C) is attributed to the different material properties assessed in each technique. However, the overall Tg behavior, the decrease in Tg with increase in drug loading in ASDs, is universally observed in all the above-mentioned techniques, which reveals their complementarity. DMA and DRS techniques are used to study the different sub-Tg transitions present in PVPVA, amorphous IND, and IND/PVPVA-based ASDs because these transitions are normally too weak or too broad for mDSC to detect. For IND/PVPVA-based ASDs, both techniques show a shift of sub-Tg transitions (or secondary relaxation peaks) toward the high-temperature region from -140 to -45 °C. Thus, this paper outlines the usage of different solid-state characterization techniques in understanding the different molecular dynamics present in the polymer, drug, and their interactions in ASDs with the integrated information obtained from individual techniques.

Carbon nanotubes (CNTs) are one of the unique and promising nanomaterials that possess plenty of applications, such as biosensors, advanced drug delivery systems and biotechnology. CNTs bind rapidly with proteins, which result in the formation of a protein coating layer known as a \"protein corona\" around the surface of the nanomaterial. This hinders their applications as a drug carrier and influences the properties of biological macromolecules. The present work focuses on studying the thermal stability and molecular level interactions of two heme proteins, hemoglobin (Hb) and myoglobin (Mb), in the presence of carboxylated functionalized multi-walled CNTs (CA-MWCNTs). Through the current study, the following steps have been taken to distinguish the biocompatibility of the hydrophilic surface CA-MWCNTs for heme proteins via a series of spectroscopic techniques and differential scanning calorimetry (DSC). UV-Visible and steady-state fluorescence spectroscopy were used to reveal changes in the aromatic amino acid residues of heme proteins upon the addition of CA-MWCNTs. Circular dichroism spectroscopy (CD) shows the alteration in the native structure of proteins in the presence of the nanomaterial. A tremendous increase in the size of the protein CA-MWCNTs system is observed in dynamic light scattering (DLS), which clearly manifests the protein corona formation. Unexpectedly, both proteins interact differently with CA-MWCNTs, which is observed in CD spectroscopy and DSC. In the presence of CA-MWCNTs, an increase in the transition temperature (Tm) was observed for Hb, while the Tm value decreases for Mb. Different interactions with proteins at the molecular scale may be the reason for this unexpected behavior. Henceforth, the present results can help in the design of the next-generation drug carrier nanomaterials with the idea of the heme protein corona formation prior to development.

Experimental techniques, such as cryo-electron microscopy, require biological samples to be recovered at cryogenic temperatures (T ≈ 100 K) with water being in an amorphous ice state. However, (bulk) water can exist in two amorphous ices at P < 1 GPa, low-density amorphous (LDA) ice at low pressures and high-density amorphous ice (HDA) at high pressures; HDA is ≈20-25% denser than LDA. While fast/plunge cooling at 1 bar brings the sample into LDA, high-pressure cooling (HPC), at sufficiently high pressure, produces HDA. HDA can also be produced by isothermal compression of LDA at cryogenic temperatures. Here, we perform classical molecular dynamics simulations to study the effects of LDA, HDA, and the LDA-HDA transformation on the structure and hydration of a small peptide, polyalanine. We follow thermodynamic paths corresponding to (i) fast/plunge cooling at 1 bar, (ii) HPC at P = 400 MPa, and (iii) compression/decompression cycles at T = 80 K. While process (i) produced LDA in the system, path (iii) produces HDA. Interestingly, the amorphous ice produced in process (ii) is an intermediate amorphous ice (IA) with properties that fall in-between those of LDA and HDA. Remarkably, the structural changes in polyalanine are negligible at all conditions studied (0-2000 MPa, 80-300 K) even when water changes among the low and high-density liquid states as well as the amorphous solids LDA, IA, and HDA. The similarities and differences in the hydration of polyalanine vitrified in LDA, IA, and HDA are described. Since the studied thermodynamic paths are suitable for the cryopreservation of biomolecules, we also study the structure and hydration of polyalanine along isobaric and isochoric heating paths, which can be followed experimentally for the recovery of cryopreserved samples. Upon heating, the structure of polyalanine remains practically unchanged. We conclude with a brief discussion of the practical advantages of (a) using HDA and IA as a cryoprotectant environment (as opposed to LDA), and (b) the use of isochoric heating as a recovery process (as opposed to isobaric heating).