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Biological drugs intended for multi-dose application require the presence of antimicrobial preservatives to avoid microbial growth. As the presence of certain preservatives has been reported to increase protein and peptide particle formation, it is essential to choose a preservative compatible with the active pharmaceutical ingredient in addition t...
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... Both empirical and statistical (DOE) screening studies with hundreds of candidate multi-dose formulations were then performed. Seven different APs found in approved parenteral products were evaluated [6,14,18], both individually and in combination, including chlorobutanol (CB), m-cresol (MC), 2-phenoxyl ethanol (2-PE), benzyl alcohol (BA), phenol, (PH) methylparaben (MP), and propylparaben (PP) [15]. Stable candidate HPV VLP multi-dose formulations included certain APs (CB, 2-PE, BA, or 2-PE + BA), while other APs (MC, PH, and parabens) displayed suboptimal stability [16]. ...
... In comparison to our results with AH-adsorbed HPV16 VLPs, mechanistic studies on the structural destabilization effects of APs used in multi-dose formulations of protein therapeutic candidates have been reported on a variety of smaller, globular proteins (in solution), including recombinant human growth hormone [21], recombinant human interleukin-1 receptor antagonist (rhIL-1ra) [20,44], recombinant human granulocyte colony stimulating factor (rhGCSF) [45], interferon alpha-2a (IFNA2), and cytochrome c [46][47][48]. In these studies, with APs other than TH, the interactions between APs and proteins were hypothesized to occur primarily through non-covalent hydrophobic forces [18,20,44,49]. Many of these APs (e.g., 2-phenoxyethanol, benzyl alcohol, phenol, methyl/propylparaben, and m-cresol) are phenolic or phenyl derivatives and are thus mostly hydrophobic in nature. ...
... Interestingly, the addition of BA increased aggregation rates and caused minor disruptions in the tertiary structures of proteins but did not significantly alter the secondary structure and overall stability of proteins [20,44,50,51]. The relatively low binding affinities of APs to these proteins permit relatively straightforward mitigation strategies such as the addition of preferentially excluded excipients (i.e., sucrose), adjusting the pH and ionic strength of the buffer, or minimizing exposure to elevated temperatures [18]. ...
During the multi-dose formulation development of recombinant vaccine candidates, protein antigens can be destabilized by antimicrobial preservatives (APs). The degradation mechanisms are often poorly understood since available analytical tools are limited due to low protein concentrations and the presence of adjuvants. In this work, we evaluate different analytical approaches to monitor the structural integrity of HPV16 VLPs adsorbed to Alhydrogel™ (AH) in the presence and absence of APs (i.e., destabilizing m-cresol, MC, or non-destabilizing chlorobutanol, CB) under accelerated conditions (pH 7.4, 50 °C). First, in vitro potency losses displayed only modest correlations with the results from two commonly used methods of protein analysis (SDS-PAGE, DSC). Next, results from two alternative analytical approaches provided a better understanding of physicochemical events occurring under these same conditions: (1) competitive ELISA immunoassays with a panel of mAbs against conformational and linear epitopes on HPV16 VLPs and (2) LC-MS peptide mapping to evaluate the accessibility/redox state of the 12 cysteine residues within each L1 protein comprising the HPV16 VLP (i.e., with 360 L1 proteins per VLP, there are 4320 Cys residues per VLP). These methods expand the limited analytical toolset currently available to characterize AH-adsorbed antigens and provide additional insights into the molecular mechanism(s) of AP-induced destabilization of vaccine antigens.
... The use of combinations of benzyl alcohol with chlorobutanol and benzyl alcohol with methylparaben may enhance antifungal activity of the formulation compared to the single use of preservatives. The combination of methylparaben and propylparaben increases the antibacterial effect of the preservative system, but also affects the solubility, and enables the protection of both phases of the emulsions [31]. It is speculated that alcohols (benzyl alcohol and pentyl alcohol) exhibit synergistic effects with benzalkonium chloride [32]. ...
The aim of this study was to indicate the type of preservatives used in selected categories of cosmetic products sold in Poland (part of the EU market) and determine the frequency of their use. The tested products consisted of 200 leave-on cosmetics, viz. body lotions (n = 100) and face creams (n = 100) and rinse-off cosmetics (n = 100) and mascaras (n = 25). The product labels of 325 adult cosmetic products from international brands were analyzed for the presence of preservatives based on the INCI compositions. The survey focused on preservatives included in Annex V of the Regulation (EC) No. 1223/2009 of the European Parliament and Council of 30 November 2009 on cosmetic products. The tested products contained 29 different preservatives belonging to eight chemical groups. Most preservatives were alcohols or their derivatives, carboxylic acids or their salts, or parabens. The most common types were phenoxyethanol, present in 198/325 (60.9%) formulations, followed by sodium benzoate, in 137 (42.2%), potassium sorbate, in 116 (35.7%), benzyl alcohol, in 76 (23.4%), and methylparaben in 33 (10.2%). Also, 33 of the 60 preservatives included in Annex V of Regulation (EC) No. 1223/2009 were not used in any of the tested preparations. In each category of products, the most common were combinations of two preservatives per single product (34.8% of all products), followed by single-preservative products (25.5%) and three-preservative products (19.4%).
... Multidose formulations are drug products packaged to permit the removal of multiple doses from a single container. The multidose format offer benefits over single-dose formats both from economic and patientcentric perspectives, as reviewed by Stroppel et al (Stroppel et al., 2023). In addition to a lower manufacturing cost, multidose formulations help to address potential issues related to dose preparation errors, drug product wastage, cost of goods for each container closure and delivery device. ...
... If gels are prepared following a straightforward freeze-drying method, cryoaerogel structures can be formulated. Due to the freezing of the gel and the subsequent sublimation to remove the solvent, a highly porous solid structure is obtained [10][11][12][13][14][15][16][17]. ...
... In a recent study, a mesoporous tablet-shaped potato starch aerogel was developed as carriers of celecoxib in order to enhance the drug solubility. The obtained data revealed that the prepared aerogels were able to improve the dissolution rate of the loaded celecoxib compared to the pure drug [14]. In another research study, using the Minitab experimental design software, the major factors for the aerogel synthesis along with optimal values of these parameters were specified. ...
In this study, a starch cryoaerogel formulation was developed as a carrier for poorly water-soluble drugs, like atorvastatin. Cryoaerogels were generated through a sol–gel method combined with a freeze-drying technique, and atorvastatin was incorporated into the obtained mesoporous systems during the solvent exchange stage. The formulated drug-loaded polymer structures were characterized in terms of their physicochemical properties, solid-state behavior, and cytotoxicity. They had a pore size of 27.56 nm and a drug loading size of 38.60%. Fourier transform infrared (FTIR) and scanning electron microscopy (SEM) analyses indicated that atorvastatin was successfully incorporated into the cryoaerogel pores. The amorphous nature of the loaded drug was confirmed via X-ray diffraction (XRD). Furthermore, after the atorvastatin incorporation into the cryogel, the volume of nitrogen adsorbed on one gram of cryoaerogel (Vm), as well as the specific surface area (aBET) were reduced. The comparison between the drug release profiles of crystalline atorvastatin and the loaded formulation of atorvastatin showed that by including the drug into the pores of the developed cryoaerogel matrix its solubility was significantly improved—the time for the dissolution of 30% pure atorvastatin (t30%) was approximately 4 h, whereas the determined t30% for the formulated cryoaerogels was only 1 h. Moreover, the data from the MTT assay illustrated that the designed cryoaerogel could be used as a safe oral atorvastatin delivery system. According to obtained results, it could be concluded that the starch cryoaerogel formulation is a promising candidate for oral delivery of poorly water-soluble therapeutic agents.
... In order to ensure sterility, multidose formulations contain antimicrobial preservatives (APs) such as, e.g., phenol, m-cresol, benzyl alcohol, thimerosal or chlorobutanol [135,136] (for a summary of antimicrobial preservative-containing peptide and protein formulations listed in the Physicians' Desk Reference, PDR, see [135]). Some of the common antimicrobial preservatives are susceptible to oxidative degradation, potentially generating radicals in pharmaceutical formulations. ...
... The exposure of benzyl alcohol to air leads to the slow formation of benzaldehyde, Ph-CHO [136]. Benzaldehyde spontaneously oxidizes to benzoic acid [137]. ...
Oxidation represents a major pathway for the chemical degradation of pharmaceutical formulations. Few specific details are available on the mechanisms that trigger oxidation reactions in these formulations, specifically with respect to the formation of free radicals. Hence, these mechanisms must be formulated based on information on impurities and stress factors resulting from manufacturing, transportation and storage. In more detail, this article focusses on autoxidation, metal-catalyzed oxidation, photo-degradation and radicals generated from cavitation as a result of mechanical stress. Emphasis is placed on probable rather than theoretically possible pathways.
Essential oils (EOs) are plant natural products typically composed of a complex mixture of molecules, many of them recognized for their antibacterial potential. EOs have different benefits in contrast to synthetic agrochemicals, like no persistence on the environment, compatibility with classical biocontrol agents and natural enemies, and no toxicity for most non-target organisms. The aim of this work was to evaluate the antibacterial effect of lemon (Citrus limon) and orange (Citrus sinensis) EOs and their combinations with mint (Mentha x piperita) or oregano (Origanum vulgare) EOs against Streptomyces scabiei, the main causal agent of the potato common scab. The EOs were characterized by GC-MS. The antibacterial activity of citrus EOs and combinations were evaluated using the broth microdilution method and the effect of the EOs on the S. scabiei envelope was analyzed by scanning electron microscopy. The lowest EO antibacterial concentrations were obtained for the combination of lemon and mint EOs, mixed in equal quantities. The application of this combination of lemon EO on S. scabiei cultures caused cell envelope damage, causing changes in the superficial hyphae anatomy and narrowing their thickness. Additionally, orange EO efficacy was enhanced in combination with oregano EO, suggesting that this binary mixture could improve their utilization as bactericidal agents against S. scabiei. These results position citrus EOs and some of their combinations as promising sustainable alternatives for potato common scab control.
Due to its involvement in skin maintenance and repair, topical administration of recombinant human growth hormone (rhGH) is an interesting strategy for therapeutic purposes. We have formulated and characterized a topical rhGH-loaded liposomal formulation (rhGH-Lip) and evaluated its safety, biological activity, and preventive role against UVB-induced skin damage. The rhGH-Lip had an average size and zeta potential of 63 nm and −33 mV, respectively, with 70 % encapsulation efficiency. The formulation was stable at 4 °C for at least one year. The SDS-PAGE and circular dichroism results showed no structural alterations in rhGH upon encapsulation. In vitro, studies in HaCaT, HFFF-2, and Ba/F3-rhGHR cell lines confirmed the safety and biological activity of rhGH-Lip. Franz diffusion cell study showed increased rhGH skin permeation compared to free rhGH. Animal studies in nude mice showed that liposomal rhGH prevented UVB-induced epidermal hyperplasia, angiogenesis, wrinkle formation, and collagen loss, as well as improving skin moisture. The results of this study show that rhGH-Lip is a stable, safe, and effective skin delivery system and has potential as an anti-wrinkle formulation for topical application. This study also provides a new method for the topical delivery of proteins and merits further investigation.
The combined effect of food preservatives – sodium benzoate and ascorbic acid on the state of monooxygenase system (MOS) components in the microsomal fraction of rat liver cells are studied in the paper. Changes in the level of cytochrome P450 and cytochrome b5, as well as the rate of reduction-oxidation of these hemoproteins in the microsomal fraction of the rat liver under conditions of administration of sodium benzoate and ascorbic acid were studied. During the experiment, rats were divided into four groups: group I – control (intact animals); group II – rats that were injected with ascorbic acid at a dose of 30 mg per kg of animal weight; group III – rats that were injected with sodium benzoate at a dose of 750 mg per kg of animal weight; group IV – rats that were injected with sodium benzoate 30 minutes before the introduction of ascorbic acid. Sodium benzoate and ascorbic acid were administered per os daily for 21 days. Animals were euthanized under light ether anesthesia on the 21st day after the start of administration of sodium benzoate and ascorbic acid. It was established that the three-week introduction of sodium benzoate into the body of animals leads to a decrease in the level of cytochrome P450, which occurs due to an increase in the rate of transition of cytochrome P450 into its inactive form P420. It was shown that along with the decrease in cytochrome P450 in the microsomal fraction of the liver, the level of cytochrome b5 decreases with a simultaneous increase in the rate of reduction and oxidation of this hemoprotein. Sodium benzoate exhibits a higher destructive effect when it is combined with ascorbic acid, which is expressed by a decrease in the level of MOS hemoproteins and may be a consequence of dangerous metabolites formation in the body - benzoic acid and benzene.
