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Introduction: Celiac disease is a common autoimmune-like enteropathy caused by an aberrant response to incompletely digested dietary gluten. Gluten immunogenic peptides including the immunodominant 33-mer are thought to be resistant to proteolytic digestion by human gastrointestinal peptidases. We developed a novel enzyme therapy approach to support gluten peptide digestion using a combination of two tandem-acting exopeptidases, AMYNOPEP, that complement the intrinsic enzymatic activity of intestinal brush border enterocytes. Methods: We evaluated the effects of AMYNOPEP supplementation on 33-mer degradation in vitro and in vivo. In a cross-over clinical study, healthy volunteers with no gastrointestinal disorders were given stable isotope (SI) labelled 33-mer peptides in the presence of varying peptide substrates and caloric loads, with and without AMYNOPEP. 33-mer degradation products (SI-labelled single amino acids) were measured in the blood plasma using LC-MS/MS. Results: AMYNOPEP achieved rapid, complete amino-to-carboxyl terminal degradation of the 33-mer in vitro, generating single amino acids and dipeptides. In healthy volunteers, AMYNOPEP supplementation significantly increased 33-mer degradation and absorption of SI-labelled amino acids even in the presence of competing substrates. Specifically, we observed a 2.8-fold increase in the Cmax of stable isotope-labelled amino acids in the presence of wheat gluten. The absorption kinetics of labelled amino acids derived from 33- mer digestion with AMYNOPEP closely resembled that of SI-labelled X-Proline dipeptides administered without enzyme supplementation, highlighting the rapid hydrolytic activity of AMYNOPEP on polypeptides. Conclusions: AMYNOPEP achieved complete degradation of the 33-mer into single amino acids and dipeptides in vitro and significantly improved 33-mer degradation kinetics in healthy volunteers, as measured by labelled amino acid detection, warranting further investigation into the potential therapeutic benefits of exopeptidase combinations for patients with gluten-related health disorders including celiac disease. [ABSTRACT FROM AUTHOR]

Abstract Background Limiting postoperative edema, pain, trismus, and infection is crucial for smooth healing. This prospective, controlled clinical trial investigated and compared the effectiveness of dexamethasone and hyaluronidase in relieving these complications. Methods In groups Ia and IIa, 8 mg of dexamethasone and 150 IU of hyaluronidase were administered following the removal of impacted teeth, respectively. The contralateral sides (groups Ib and IIb) were determined as control groups. Edema, pain, trismus, and infection were clinically evaluated on the 1st, 2nd, 3rd, and 7th postoperative days. Results 60 patients were enrolled in the study. Hyaluronidase provided significantly more edema relief than dexamethasone on the 1st, 2nd, 3rd, and 7th postoperative days (P = 0.031, 0.002, 0.000, and 0.009, respectively). No statistical difference was found between dexamethasone and hyaluronidase in VAS and rescue analgesic intake amount values for all time points. Hyaluronidase was more effective in reducing trismus than dexamethasone on the 2nd and 3rd postoperative days (P = 0.029, 0.024, respectively). Neither of the agents significantly increased the postoperative infection rate. Conclusions Hyaluronidase can be selected when postoperative excessive edema and trismus are anticipated. Dexamethasone may be a cost–effective option if postoperative pain control is merely targeted. Trial registration This trial was registered in the Clinical Trials Protocol Registration and Results System (ClinicalTrials.gov identifier number: NCT05466604) on 20/07/2022.

Background: Limiting postoperative edema, pain, trismus, and infection is crucial for smooth healing. This prospective, controlled clinical trial investigated and compared the effectiveness of dexamethasone and hyaluronidase in relieving these complications. Methods: In groups Ia and IIa, 8 mg of dexamethasone and 150 IU of hyaluronidase were administered following the removal of impacted teeth, respectively. The contralateral sides (groups Ib and IIb) were determined as control groups. Edema, pain, trismus, and infection were clinically evaluated on the 1st, 2nd, 3rd, and 7th postoperative days. Results: 60 patients were enrolled in the study. Hyaluronidase provided significantly more edema relief than dexamethasone on the 1st, 2nd, 3rd, and 7th postoperative days (P = 0.031, 0.002, 0.000, and 0.009, respectively). No statistical difference was found between dexamethasone and hyaluronidase in VAS and rescue analgesic intake amount values for all time points. Hyaluronidase was more effective in reducing trismus than dexamethasone on the 2nd and 3rd postoperative days (P = 0.029, 0.024, respectively). Neither of the agents significantly increased the postoperative infection rate. Conclusions: Hyaluronidase can be selected when postoperative excessive edema and trismus are anticipated. Dexamethasone may be a cost–effective option if postoperative pain control is merely targeted. Trial registration: This trial was registered in the Clinical Trials Protocol Registration and Results System (ClinicalTrials.gov identifier number: NCT05466604) on 20/07/2022. [ABSTRACT FROM AUTHOR]

In recent years, enzyme therapy strategies have rapidly evolved to catalyze essential biochemical reactions with therapeutic potential. These approaches hold particular promise in addressing rare genetic disorders, cancer treatment, neurodegenerative conditions, wound healing, inflammation management, and infectious disease control, among others. There are several primary reasons for the utilization of enzymes as therapeutics: their substrate specificity, their biological compatibility, and their ability to generate a high number of product molecules per enzyme unit. These features have encouraged their application in enzyme replacement therapy where the enzyme serves as the therapeutic agent to rectify abnormal metabolic and physiological processes, enzyme prodrug therapy where the enzyme initiates a clinical effect by activating prodrugs, and enzyme dynamic or starving therapy where the enzyme acts upon host substrate molecules. Currently, there are >20 commercialized products based on therapeutic enzymes, but approval rates are considerably lower than other biologicals. This has stimulated nanobiotechnology in the last years to develop nanoparticle-based solutions that integrate therapeutic enzymes. This approach aims to enhance stability, prevent rapid clearance, reduce immunogenicity, and even enable spatio-temporal activation of the therapeutic catalyst. This comprehensive review delves into emerging trends in the application of therapeutic enzymes, with a particular emphasis on the synergistic opportunities presented by incorporating enzymes into nanomaterials. Such integration holds the promise of enhancing existing therapies or even paving the way for innovative nanotherapeutic approaches. [Display omitted] • Enzyme therapies offer new advanced treatments for a wide range of medical conditions. • Enzyme specificity and high product yield are crucial for prodrug activation, replacement, dynamic and starving therapies. • Combining enzymes and nanomaterials enables precise targeting and spatio-temporal remote control of these therapies. [ABSTRACT FROM AUTHOR]

Enzyme‐based therapy has garnered significant attention for its current applications in various diseases. Despite the notable advantages associated with the use of enzymes as therapeutic agents, that could have high selectivity, affinity, and specificity for the target, their application faces challenges linked to physico‐chemical and pharmacological properties. These limitations can be addressed through the encapsulation of enzymes in nanoplatforms as a comprehensive solution to mitigate their degradation, loss of activity, off‐target accumulation, and immunogenicity, thus enhancing bioavailability, therapeutic efficacy, and circulation time, thereby reducing the number of administrations, and ameliorating patient compliance. The exploration of novel nanomedicine‐based enzyme therapeutics for the treatment of challenging diseases stands as a paramount goal in the contemporary scientific landscape, but even then it is often not enough. Combining an enzyme with another therapeutic (e.g., a small molecule, another enzyme or protein, a monoclonal antibody, or a nucleic acid) within a single nanocarrier provides innovative multidrug‐integrated therapy and ensures that both the actives arrive at the target site and exert their therapeutic effect, leading to synergistic action and superior therapeutic efficacy. Moreover, this strategic approach could be extended to gene therapy, a field that nowadays has gained increasing attention, as enzymes acting at genomic level and nucleic acids may be combined for synergistic therapy. This multicomponent therapeutic approach opens opportunities for promising future developments. This article is categorized under:Therapeutic Approaches and Drug Discovery > Nanomedicine for Neurological DiseaseTherapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic DiseaseTherapeutic Approaches and Drug Discovery > Emerging Technologies [ABSTRACT FROM AUTHOR]

Abstract It is recognized as a promising therapeutic strategy for cocaine use disorder to develop an efficient enzyme which can rapidly convert cocaine to physiologically inactive metabolites. We have designed and discovered a series of highly efficient cocaine hydrolases, including CocH5-Fc(M6) which is the currently known as the most efficient cocaine hydrolase with both the highest catalytic activity against (−)-cocaine and the longest biological half-life in rats. In the present study, we characterized the time courses of protein appearance, pH, structural integrity, and catalytic activity against cocaine in vitro and in vivo of a CocH5-Fc(M6) bulk drug substance produced in a bioreactor for its in vitro and in vivo stability after long-time storage under various temperatures (− 80, − 20, 4, 25, or 37 °C). Specifically, all the tested properties of the CocH5-Fc(M6) protein did not significantly change after the protein was stored at any of four temperatures including − 80, − 20, 4, and 25 °C for ~ 18 months. In comparison, at 37 °C, the protein was less stable, with a half-life of ~ 82 days for cocaine hydrolysis activity. Additionally, the in vivo studies further confirmed the linear elimination PK profile of CocH5-Fc(M6) with an elimination half-life of ~ 9 days. All the in vitro and in vivo data on the efficacy and stability of CocH5-Fc(M6) have consistently demonstrated that CocH5-Fc(M6) has the desired in vitro and in vivo stability as a promising therapeutic candidate for treatment of cocaine use disorder.

Chronic endometritis "from pathogenesis to therapy" still remains the most contrarian issue of modern gynaecology. The lack of a unified pathogenetic concept as well as diagnostic and therapeutic algorithms continue to support a multitude of scientific discussions around this unique nosology. Latent subclinical endometrial inflammatory changes are associated with the realization of reproductive failures, including repeated implantation failures and early reproductive losses. To date, a sufficient amount of scientific data has been accumulated demonstrating high efficacy and safety of antifibrotic therapy with the use of azoximer bovgialuronidase (Longidaza) in the complex treatment of chronic endometritis.

Abstract Polymicrobial communities lead to worsen the wound infections, due to mixed biofilms, increased antibiotic resistance, and altered virulence production. Promising approaches, including enzymes, may overcome the complicated condition of polymicrobial infections. Therefore, this study aimed to investigate Staphopain A-mediated virulence and resistance alteration in an animal model of Staphylococcus aureus and Pseudomonas aeruginosa co-infection. S. aureus and P. aeruginosa were co-cultured on the L-929 cell line and wound infection in an animal model. Then, recombinant staphopain A was purified and used to treat mono- and co-infections. Following the treatment, changes in virulence factors and resistance were investigated through phenotypic methods and RT-PCR. Staphopain A resulted in a notable reduction in the viability of S. aureus and P. aeruginosa. The biofilm formed in the wound infection in both animal model and cell culture was disrupted remarkably. Moreover, the biofilm-encoding genes, quorum sensing regulating genes, and virulence factors (hemolysin and pyocyanin) controlled by QS were down-regulated in both microorganisms. Furthermore, the resistance to vancomycin and doripenem decreased following treatment with staphopain A. According to this study, staphopain A might promote wound healing and cure co-infection. It seems to be a promising agent to combine with antibiotics to overcome hard-to-cure infections.

It is recognized as a promising therapeutic strategy for cocaine use disorder to develop an efficient enzyme which can rapidly convert cocaine to physiologically inactive metabolites. We have designed and discovered a series of highly efficient cocaine hydrolases, including CocH5-Fc(M6) which is the currently known as the most efficient cocaine hydrolase with both the highest catalytic activity against (−)-cocaine and the longest biological half-life in rats. In the present study, we characterized the time courses of protein appearance, pH, structural integrity, and catalytic activity against cocaine in vitro and in vivo of a CocH5-Fc(M6) bulk drug substance produced in a bioreactor for its in vitro and in vivo stability after long-time storage under various temperatures (− 80, − 20, 4, 25, or 37 °C). Specifically, all the tested properties of the CocH5-Fc(M6) protein did not significantly change after the protein was stored at any of four temperatures including − 80, − 20, 4, and 25 °C for ~ 18 months. In comparison, at 37 °C, the protein was less stable, with a half-life of ~ 82 days for cocaine hydrolysis activity. Additionally, the in vivo studies further confirmed the linear elimination PK profile of CocH5-Fc(M6) with an elimination half-life of ~ 9 days. All the in vitro and in vivo data on the efficacy and stability of CocH5-Fc(M6) have consistently demonstrated that CocH5-Fc(M6) has the desired in vitro and in vivo stability as a promising therapeutic candidate for treatment of cocaine use disorder. [ABSTRACT FROM AUTHOR]

Polymicrobial communities lead to worsen the wound infections, due to mixed biofilms, increased antibiotic resistance, and altered virulence production. Promising approaches, including enzymes, may overcome the complicated condition of polymicrobial infections. Therefore, this study aimed to investigate Staphopain A-mediated virulence and resistance alteration in an animal model of Staphylococcus aureus and Pseudomonas aeruginosa co-infection. S. aureus and P. aeruginosa were co-cultured on the L-929 cell line and wound infection in an animal model. Then, recombinant staphopain A was purified and used to treat mono- and co-infections. Following the treatment, changes in virulence factors and resistance were investigated through phenotypic methods and RT-PCR. Staphopain A resulted in a notable reduction in the viability of S. aureus and P. aeruginosa. The biofilm formed in the wound infection in both animal model and cell culture was disrupted remarkably. Moreover, the biofilm-encoding genes, quorum sensing regulating genes, and virulence factors (hemolysin and pyocyanin) controlled by QS were down-regulated in both microorganisms. Furthermore, the resistance to vancomycin and doripenem decreased following treatment with staphopain A. According to this study, staphopain A might promote wound healing and cure co-infection. It seems to be a promising agent to combine with antibiotics to overcome hard-to-cure infections. [ABSTRACT FROM AUTHOR]

Glucose oxidase (GOx)-based enzyme therapeutics are potential alternatives for colorectal cancer (CRC) treatment via glucose consumption and accumulation of hydrogen peroxide (H 2 O 2). Given that H 2 O 2 can be eliminated by cytoprotective autophagy, autophagy inhibitors that can interrupt autolysosome-induced H 2 O 2 elimination are promising combination drugs of GOx. Here, we developed a multifunctional biomimetic nanocarrier for effective co-delivery of an autophagy inhibitor-chloroquine phosphate (CQP) and GOx to exert their synergistic effect by irreversibly upregulating intracellular reactive oxygen species (ROS) levels. Poly (D, l -lactide- co -glycolide) (PLGA) nanoparticles (NPs) were used to encapsulate both GOx and CQP using a W/O/W multi-emulsion method. Calcium phosphate (CaP) was used to "fix" CQP to GOx in the internal water phase, where it served as a pH-sensitive unit to facilitate intracellular drug release. Folic acid-modified red blood cell membranes (FR) were used to camouflage the GOx/CQP/CaP encapsulated PLGA NPs (referred to as PLGA/GCC@FR). In an AOM/DSS-induced CRC mouse model, PLGA/GCC@FR exhibited improved antitumor effects, in which the number of tumor nodes were only a quarter of that in the free drug combination group. The enhanced therapeutic effects of PLGA/GCC@FR were attributed to the prolonged tumor retention which was verified by both dynamic in vivo imaging and drug biodistribution. This multifunctional biomimetic nanocarrier facilitated combined enzyme therapeutics by depleting glucose and augmenting intracellular ROS levels in tumor cells, which exerted a synergistic inhibitory effect on tumor growth. Therefore, this study proposed a novel strategy for the enhancement of combined enzyme therapeutics, which provided a promising method for effective CRC treatment. [Display omitted] • CQP was fixed to GOx via CaP formation to facilitate co-encapsulation by PLGA NPs. • PBA-PEG-FA was inserted on RBCm to realize "right-side-out" coating and CRC targeting. • Glucose consumption and ROS accumulation mediated CRC tumor regression. [ABSTRACT FROM AUTHOR]

The purpose of this study is to evaluate the clinical and laboratory manifestations of children with mucopolysaccharidosis type 6 before and after enzyme therapy. In this quasi-experimental study, 8 patients with MPS-6 referred to the pediatric endocrinology department of Imam Reza Hospital in Mashhad were followed up for 12 months. The level of urinary glycosaminoglycan was measured to check the response to the treatment. The range of motion of the shoulder and elbow joints was evaluated using a goniometer, and abdominal ultrasound was performed to check the size of the liver and spleen in the midclavicular line. The 6-minute walking test and the 3-minute stair climbing test were performed for the patients at the mentioned times. The height and weight of the patients were also measured, and echocardiography was performed. Then patients underwent weekly enzyme treatment. One of the patients (seventh patient) was excluded from the study. Patients were treated with enzyme from the beginning of the study. The patients were evaluated at 12 months later. Statistical analysis showed that changes in urinary GAG level, height, weight, changes in 6-minute walk and range of motion (extension and flexion) of the shoulder were significant (P<0.05). Changes in liver and spleen size compared to height, climbing stairs, corneal opacity and heart changes after 12 months of enzyme therapy were not significant. It is suggested that even though this method of treatment is not definitive, that can be continued to improve the current condition of the patient. [ABSTRACT FROM AUTHOR]

One of the serious intra-abdominal complications after surgery is intestinal obstruction of adhesion origin (IOAO). Mortality during postoperative intestinal obstruction is 16.2–52.5 %. Aim of the study – laparoscopic diagnosis of IOAO in the early postoperative period and optimization of adhesiolysis.Material and methods. In the early postoperative period 70 patients with IOAO of upper gastrointestinal tract were examined and treated; laparoscopic operation was performed to 46 patients (the main group), an open method (laparotomy) – to 24 patients (the control group).Results and discussion. It was revealed that upper-middle and lower-middle incision laparotomies are more likely to cause intestinal obstruction. The sensitivity of preoperative ultrasound diagnosis of movable visceroparietal adhesions of small intestinal loops is 94.6 %, and the sensitivity of laparoscopic diagnosis is 99.2 %. When using minimally invasive technology, the frequency of intraoperative (4 (8.7 %) and 8 (33.3 %), respectively, p < 0.05) and postoperative complications (5 (10.9 %) and 13 (54.1 %), p < 0.05), mortality (0 and 4 (16.6 %), p < 0.05) and length of stay in hospital (5.7 (3–8) and 14.3 (10–17) days, p < 0,05) was less compared to laparotomy.Conclusions. Using of laparoscopic adhesiolysis and anti-adhesion barrier is appropriate in patients with I-II grade adhesion process. In cases where technical difficulties arise during laparoscopic adhesiolysis, it is more pathogenetically justified to separate adhesions by passing through a mini-laparotomy incision. If this is not possible, it is advisable to perform complex anti-adhesion measures, including conversion to laparotomy and systemic enzymotherapy after adhesiolysis.

Summary: Background: Arginase 1 Deficiency (ARG1-D) is a rare debilitating, progressive, inherited, metabolic disease characterized by marked increases in plasma arginine (pArg) and its metabolites, with increased morbidity, substantial reductions in quality of life, and premature mortality. Effective treatments that can lower arginine and improve clinical outcomes is currently lacking. Pegzilarginase is a novel human arginase 1 enzyme therapy. The present trial aimed to demonstrate efficacy of pegzilarginase on pArg and key mobility outcomes. Methods: This Phase 3 randomized, double-blind, placebo-controlled, parallel-group clinical trial (clinicaltrials.gov NCT03921541, EudraCT 2018-004837-34), randomized patients with ARG1-D 2:1 to intravenously/subcutaneously once-weekly pegzilarginase or placebo in conjunction with their individualized disease management. It was conducted in 7 countries; United States, United Kingdom, Canada, Austria, France, Germany, Italy. Primary endpoint was change from baseline in pArg after 24 weeks; key secondary endpoints were change from baseline at Week 24 in Gross Motor Function Measure part E (GMFM-E) and 2-min walk test (2MWT). Full Analysis Set was used for the analyses. Findings: From 01 May 2019 to 29 March 2021, 32 patients were enrolled and randomized (pegzilarginase, n = 21; placebo, n = 11). Pegzilarginase lowered geometric mean pArg from 354.0 μmol/L to 86.4 μmol/L at Week 24 vs 464.7 to 426.6 μmol/L for placebo (95% CI: −67.1%, −83.5%; p

Therapeutic enzymes present excellent opportunities for the treatment of human disease, modulation of metabolic pathways and system detoxification. However, current use of enzyme therapy in the clinic is limited as naturally occurring enzymes are seldom optimal for such applications and require substantial improvement by protein engineering. Engineering strategies such as design and directed evolution that have been successfully implemented for industrial biocatalysis can significantly advance the field of therapeutic enzymes, leading to biocatalysts with new‐to‐nature therapeutic activities, high selectivity, and suitability for medical applications. This minireview highlights case studies of how state‐of‐the‐art and emerging methods in protein engineering are explored for the generation of therapeutic enzymes and discusses gaps and future opportunities in the field of enzyme therapy. [ABSTRACT FROM AUTHOR]

A significant proportion of lower-grade glioma as well as many other types of human cancers are associated with neomorphic mutations in IDH1/2 genes (mIDH1/2). These mutations lead to an aberrant accumulation of 2- hydroxyglutarate (2-HG). Interestingly, even cancers without mIDH1/2 can exhibit increased levels of 2-HG due to factors like hypoxia and extracellular acidity. Mounting evidence demonstrates that 2-HG competitively inhibits aketoglutarate dependent enzymes, such as JmjC-domain-containing histone demethylases (JHDMs), ten-eleven translocation enzymes (TETs), and various dioxygenases (e.g., RNA m6A demethylases and prolyl hydroxylases). Consequently, the hypermethylation of DNA, RNA, and histones, and the abnormal activities of hypoxia-inducible factors (HIFs) have profound impacts on the establishment of cancer metabolism and microenvironment, which promote tumor progression. This connection between the oncometabolite 2- HG and glioma holds crucial implications for treatments targeting this disease. Here, I hypothesize that an ectopic introduction of a bacterial 2-hydroxyglutarate synthase (2-HG synthase) enzyme into cancer cells with 2-HG accumulation could serve as a promising enzyme therapy for glioma and other types of cancers. While absent in human metabolism, 2-HG synthase in bacterial species catalyzes the conversion of 2-HG into propionyl-CoA and glyoxylate, two metabolites that potentially possess anti-tumor effects. For a broad spectrum of human cancers with 2-HG accumulation, 2-HG synthase-based enzyme therapy holds the potential to not only correct 2-HG induced cancer metabolism but also transform an oncometabolite into metabolic challenges within cancer cells. [ABSTRACT FROM AUTHOR]

Therapeutic enzymes used for genetic disorders or metabolic diseases oftentimes suffer from suboptimal pharmacokinetics and stability. Nanodelivery systems have shown considerable promise for improving the performance of enzyme therapies. Here, we develop a cell membrane-camouflaged metal-organic framework (MOF) system with enhanced biocompatibility and functionality. The MOF core can efficiently encapsulate enzymes while maintaining their bioactivity. After the introduction of natural cell membrane coatings, the resulting nanoformulations can be safely administered in vivo. The surface receptors on the membrane can also provide additional functionalities that synergize with the encapsulated enzyme to target disease pathology from multiple dimensions. Employing uricase as a model enzyme, we demonstrate the utility of this approach in multiple animal disease models. The results support the use of cell membrane-coated MOFs for enzyme delivery, and this strategy could be leveraged to improve the usefulness of enzyme-based therapies for managing a wide range of important human health conditions.

Cystathionine beta‐synthase (CBS)‐deficient homocystinuria (HCU) is the most common inborn error of sulfur amino acid metabolism. The pyridoxine non‐responsive form of the disease manifests itself by massively increasing plasma and tissue concentrations of homocysteine, a toxic intermediate of methionine metabolism that is thought to be the major cause of clinical complications including skeletal deformities, connective tissue defects, thromboembolism and cognitive impairment. The current standard of care involves significant dietary interventions that, despite being effective, often adversely affect quality of life of HCU patients, leading to poor adherence to therapy and inadequate biochemical control with clinical complications. In recent years, the unmet need for better therapeutic options has resulted in development of novel enzyme and gene therapies and exploration of pharmacological approaches to rescue CBS folding defects caused by missense pathogenic mutations. Here, we review scientific evidence and current state of affairs in development of recent approaches to treat HCU. [ABSTRACT FROM AUTHOR]

We aimed to determine the relationship of Lyso-Gb1 levels, bone biomarkers, and mutation findings with bone marrow burden (BMB) scores.

As well known, cocaine induces stimulant effects and dopamine transporter (DAT) trafficking to the plasma membrane of dopaminergic neurons. In the present study, we examined cocaine-induced hyperactivity along with cocaine-induced DAT trafficking and the recovery rate of the dopaminergic system in female rats in comparison with male rats, demonstrating interesting gender differences. Female rats are initially more sensitive to cocaine than male rats in terms of both the DAT trafficking and hyperactivity induced by cocaine. Particularly, intraperitoneal (i.p.) administration of 5 mg/kg cocaine induced significant hyperactivity and DAT trafficking in female rats but not in male rats. After repeated cocaine exposures (i.e., i.p. administration of 20 mg/kg cocaine every other day from Day 0 to Day 32), cocaine-induced hyperactivity in female rats gradually became a clear pattern of two phases, with the first phase of the hyperactivity lasting for only a few minutes and the second phase lasting for over an hour beginning at ~30 min, which is clearly different from that of male rats. It has also been demonstrated that the striatal DAT distribution of female rats may recover faster than that of male rats after multiple cocaine exposures. Nevertheless, despite the remarkable gender differences, our recently developed long-acting cocaine hydrolase, known as CocH5-Fc(M6), can similarly and effectively block cocaine-induced DAT trafficking and hyperactivity in both male and female rats. [ABSTRACT FROM AUTHOR]

Pseudoxanthoma elasticum (PXE) is a genetic multisystem ectopic calcification disorder caused by inactivating mutations in the ABCC6 gene encoding ABCC6, a hepatic efflux transporter. ABCC6-mediated ATP secretion by the liver is the main source of a potent endogenous calcification inhibitor, plasma inorganic pyrophosphate (PPi); the deficiency of plasma PPi underpins PXE. Recent studies demonstrated that INZ-701, a recombinant human ENPP1 that generates PPi and is now in clinical trials, restored plasma PPi levels and prevented ectopic calcification in the muzzle skin of Abcc6 -/- mice. This study examined the pharmacokinetics, pharmacodynamics, and potency of a new ENPP1-Fc isoform, BL-1118, in Abcc6 -/- mice. When Abcc6 -/- mice received a single subcutaneous injection of BL-1118 at 0.25, 0.5, or 1 mg/kg, they had dose-dependent elevations in plasma ENPP1 enzyme activity and PPi levels, with an enzyme half-life of approximately 100 h. When Abcc6 -/- mice were injected weekly from 5 to 15 weeks of age, BL-1118 dose-dependently increased steady-state plasma ENPP1 activity and PPi levels and significantly reduced ectopic calcification in the muzzle skin and kidneys. These results suggest that BL-1118 is a promising second generation enzyme therapy for PXE, the first generation of which is currently in clinical testing., Competing Interests: Declaration of interests D.T.B. and P.R.S. are inventors on patents owned by Yale University for therapeutics treating ENPP1 deficiency. D.T.B. is an equity holder and receives research and consulting support from Inozyme Pharma, Inc., (Copyright © 2024 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)

Aims: Antibiotic resistance including multidrug resistance (MDR) is a negative symbol to the human health system because it loses the capability to treat infections. Unfortunately, the available antibiotics do not show an effective therapeutic response against bacterial infections. In the situation of global antibiotic unresponsiveness, enzymatic therapy especially in combinatorial form seems an effective approach to control bacterial infection and combat resistance. The article is important because it focuses on combinatorial enzymatic therapy that has multiple properties (effective antibacterial performances, antibiofilm capacity, immunomodulators, targeted actions, synergistic actions, multiple targeting, and resistance-proof properties) and can address antibiotic resistance effectively., Materials and Methods: We searched the related topics with Pubmed, Scopus, and Google Scholar databases and finally 73 relevant papers were reviewed in detail and cited in this article., Key Findings: Discusses properties of combinatorial therapeutic enzymes made it an accomplished means over antibiotic therapy. This article discusses the need for combinatorial enzymatic therapy against bacterial infection, its distinguished features, and properties with multi-mechanistic antibacterial action. It discussed the European Medicine Agency and Food and Drug Administration-approved therapeutic enzymes (antibacterial and antibiofilm)., Significance: This article provided the possible combination of the enzyme that may be used as an antibacterial agent along with limitations and future scope of combinatorial antibacterial enzymatic agents. This article could draw the attention of researchers to combinatorial therapeutic enzymatic molecules as effective and futuristic therapy to overcome the problem of multiple antibiotic resistance in bacteria., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Introduction: Cellulite, also known as edematous fibrosclerotic panniculopathy (EFP), affects up to 90% of women and has a significant aesthetic impact. EFP is a multifactorial condition characterized by local circulatory changes, increased adipose tissue thickness, and a fibrotic response involving thick collagen bundles and septa, driven by local hypoxia. Although numerous treatments exist, their effects are typically temporary. This study evaluates the outcomes of four patients with EFP treated using a combined recombinant enzymatic therapy consisting of a lyase, lipase, and collagenase., Methods: A standardized protocol involving injections of a combined enzyme solution (pbserum Medium™) was administered to the lower limbs in three separate sessions. Pre- and post-treatment photographs were collected for comparative analysis., Results: All four patients showed improvements in skin appearance and fibrosis, with no systemic or local adverse events reported., Conclusions: We propose that a treatment strategy targeting the edematous, adipose, and fibrotic components of EFP may offer an economical and pathogenic-based approach for managing this condition in affected women.

Neuronal ceroid lipofuscinosis type 2 (CLN2) disease is a rare, lysosomal storage disorder that causes pediatric onset neurodegenerative disease. It is characterized by mutations in the TPP1 gene. Symptoms begin between 2 and 4 years of age with loss of previously acquired motor, cognitive, and language abilities. Cerliponase alfa, a recombinant human TPP1 enzyme, is the only approved therapy. We report the first presymptomatic cerliponase alfa intraventricular treatment in a familial case of CLN2 related to a classical TPP1 variant. Sister 1 presented with motor, cognitive, and language decline and progressive myoclonic epilepsy since the age of 3 years, evolved with severe diffuse encephalopathy, received no specific treatment, and died at 11 years. Sister 2 had a CLN2 presymptomatic diagnosis and has been treated with cerliponase since she was 12 months old. She is now 6 years 8 months and has no CLN2 symptom except one generalized seizure 1 year ago. No serious adverse event has occurred. Repeated Wechsler Preschool and Primary Scale of Intelligence, Fourth Edition standardized index scores are heterogeneous in the extremely low to low average ranges. Mean length of utterances, a global index of sentence complexity, showed a delay, but a gradual improvement. The reported case enhances the major contribution of presymptomatic diagnosis and significant middle-term treatment benefit for patients with CLN2., (© 2024 The Authors. European Journal of Neurology published by John Wiley & Sons Ltd on behalf of European Academy of Neurology.)

Relevance. The variety of functional changes in internal organs characteristicofgeriatricage necessitates the search for drugs that can exert a multifaceted effect on various links in the pathogenesis of chronic pancreatitis (CP) in comorbidity with the most common diseases of the elderly. Purpose. Investigation of the effectiveness of the inclusion of a systemic enzyme therapy drug in the complex treatment of chronic pancreatitis in elderly patients. Materials and methods. We examined 77 patients with CP over the age of 66 years, the average age (71.3 ± 1.4) g. Group I of patients with CP (23 patients) - took aconventional therapeutic complex (MC) for three months: pantoprazole 40 mg on an empty stomach according to requirement, continuous enzyme replacement therapy with pure pancreatin in an adequate dose with each meal, prokinetics / or antispasmodics if necessary. Group II consisted of 34 patients with CP (MC + W), in addition to LK, the drug of systemic enzyme therapy (SET) Wobenzym, 5 tablets, was taken. three times a day for three months. The control group consisted of 20 patients without signs of pathology of the digestive system. The parameters of lipid metabolism were determined by the enzymatic-colorimetric method. The assessment of the depth of exocrine pancreatic insufficiency (PI) was determined by the level of fecal α-elastase (FαE) by enzyme-linked immunosorbentassay using standard BIOSERVELASTASE 1-ELISA kits. Statistical analysis was performed using Excel and the statistical package Statistica v. 5.0. Results. Additional inclusion of the drug SET (Wobenzym) in the generally accepted MC promoted a significant improvement in lipid profile parameters in elderly patients with CP, as well as a reliable correction of the excretory function of the P by the level of FαE (from the level of moderate to mild insufficiency). Conclusions. 1) The inclusion of Wobenzym in the general treatment complex contributed toa decrease in the levels of TC, βLP, TG and LDLP by 15.4%, 19.8%, 32.0% and 33.9%, respectively (p 0.05).

IntroductionGene-directed enzyme prodrug therapy (GDEPT) employing the cytosine deaminase (CD) gene, which encodes an enzyme that converts the nontoxic agent 5-fluorocytosine (5-FC) into the chemotherapeutic drug 5-fluorouracil (5-FU), has shown promise both in experimental animals and in clinical trials. Nevertheless, with the transfection systems available presently the percentage of tumor cells incorporating the desired gene is usually too low for successful therapy. We have examined the ability of photodynamic therapy (PDT) to enhance the efficacy of the metabolites, converted from 5-FC by CD gene transfected rat glioma cells.MethodsHybrid tumor cell spheroids consisting of CD poitive and CD negative F98 glioma cells in varying ratios were used as in vitro tumor models. PDT was performed with the photosensitizer AlPcS2a and λ=670nm laser irradiance, both before and after confrontation with 5-FC.ResultsPDT increased the toxicity of 5-FU either as pure drug or derived from monolayers of CD positive cells chalanged with 5-FC. PDT in combination with 5-FC resulted in a significantly enhanced inhibition of hybrid spheroid growth compared to non light treated controls. This was the case even at tumor to producer cell ratios as high as 40:1.ConclusionThe results of the present study show that GDEPT and PDT interact in a synergistic manner over a range of prodrug concentration and tumor to transfected cell ratios. The degree of synergy was significant regardless if PDT treatment was given before or after 5-FC administration. The highest degree of interaction was observed though, when PDT was delivered prior to prodrug exposure.

Background and purpose: Data on interruption of enzyme replacement therapy (ERT) are scarce in late onset Pompe disease. Due to the COVID‐19 crisis, eight neuromuscular reference centers in France were obligated to stop the treatment for 31 patients. Methods: We collected the motor and respiratory data from our French registry, before COVID‐19 and at treatment restart. Results: In 2.2 months (mean), patients showed a significant deterioration of 37 m (mean) in the 6‐min walk test and a loss of 210 ml (mean) of forced vital capacity, without ad integrum restoration after 3 months of ERT restart. Conclusions: This national study based on data from the French Pompe Registry shows that the interruption of ERT, even as short as a few months, worsens Pompe patients' motor and respiratory function. [ABSTRACT FROM AUTHOR]

Enzyme therapy has important implications for the treatment of metabolic disorders and biological detoxification. It remains challenging to prepare enzymatic nanoreactors with high therapeutic efficiency and low emission of cytotoxic reaction intermediates. Here, we propose a novel strategy for the preparation of enzymes-loaded polypeptide microcapsules (EPM) with concentrically encapsulated enzymes to achieve higher cascade reaction rates and minimal emission of cytotoxic intermediates. Mesoporous silica spheres (MSS) are used as a highly porous matrix to efficiently load a therapeutic enzyme (glucose oxidase, GOx), and a layer-by-layer (LbL) assembly strategy is employed to assemble the scavenging enzyme (catalase) and polyelectrolyte multilayers on the MSS surface. After removal of the MSS, a concentrically encapsulated EPM is obtained with the therapeutic enzyme encapsulated inside the capsule, and the scavenging enzyme immobilized in the polypeptide multilayer shell. Performance of the concentrically encapsulated GOx-catalase capsules is investigated for synergistic glucose metabolism disturbance correction and cytotoxic intermediate H2O2 clearance. The results show that the EPM can simultaneously achieve 99% H2O2 clearance and doubled glucose consumption rate. This strategy can be extended to the preparation of other dual- or multi-enzyme therapeutic nanoreactors, showing great promise in the treatment of metabolic disorders. [ABSTRACT FROM AUTHOR]

Intestinal colic, food allergies are one of the common causes of the initial treatment of children in their first year of life in outpatient practice. Gastrointestinal allergy is a lesion of the gastrointestinal tract of an allergic nature, is in second place among the pathologies associated with food allergies, where allergy to cow’s milk proteins leads. In recent years, the diagnosis of lactase deficiency, unreasonably leads to an unreasonable transfer of the child to artificial curing and the appointment of lactose-free mixtures. In some patients with gastrointestinal manifestations, secondary LN is quite often formed. The article discusses the clinical forms of LN and GA. Undigested lactose increases the cell-mediated pro-inflammatory processes in the body, which, in turn, leads to the stimulation of the development of inflammatory and allergic processes in children. The possibilities and effectiveness of concomitant diet therapy are discussed, including using replacement therapy with the enzyme lactase, which eliminates the main cause of childhood colic, helps the absorption of breast milk or mixture, thus providing babies with the necessary nutrients for physiological development. The article presents results of the clinical observation carried out in a group of 20 full term babies aged 2–5 months with secondary lactase deficiency in the setting of gastrointestinal allergy with positive clinical response: highly quick control of symptoms of intestinal dyspepsia, improvement in stool frequency and consistency were detected on Day 7 from the start of enzyme therapy in 85% of children. Symptoms of intestinal colic (95%), flatulence (80%) were also reversed, regurgitation decreased in frequency and volume (85%) by the end of Week 1 of the therapy and completely disappeared by the end of Week 2. Lab tests confirmed the clinical efficacy of the therapy. A significant improvement in coprogram indicators was identified in 80% of children by the end of Week 2 from the start of therapy. The researchers found out that the carbohydrate content in the coprogram decreased by 1.8 times from 1.3% ± 0.5 to 0.7% ± 0.5 (p

Enzyme-based therapy has garnered significant attention for its current applications in various diseases. Despite the notable advantages associated with the use of enzymes as therapeutic agents, that could have high selectivity, affinity, and specificity for the target, their application faces challenges linked to physico-chemical and pharmacological properties. These limitations can be addressed through the encapsulation of enzymes in nanoplatforms as a comprehensive solution to mitigate their degradation, loss of activity, off-target accumulation, and immunogenicity, thus enhancing bioavailability, therapeutic efficacy, and circulation time, thereby reducing the number of administrations, and ameliorating patient compliance. The exploration of novel nanomedicine-based enzyme therapeutics for the treatment of challenging diseases stands as a paramount goal in the contemporary scientific landscape, but even then it is often not enough. Combining an enzyme with another therapeutic (e.g., a small molecule, another enzyme or protein, a monoclonal antibody, or a nucleic acid) within a single nanocarrier provides innovative multidrug-integrated therapy and ensures that both the actives arrive at the target site and exert their therapeutic effect, leading to synergistic action and superior therapeutic efficacy. Moreover, this strategic approach could be extended to gene therapy, a field that nowadays has gained increasing attention, as enzymes acting at genomic level and nucleic acids may be combined for synergistic therapy. This multicomponent therapeutic approach opens opportunities for promising future developments. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Neurological Disease Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease Therapeutic Approaches and Drug Discovery > Emerging Technologies., (© 2024 Wiley Periodicals LLC.)

Benzoylecgonine (BZE) is the major toxic metabolite of cocaine and is responsible for the long‐term cocaine‐induced toxicity owing to its long residence time in humans. BZE is also the main contaminant following cocaine consumption. Here, we identified the bacterial cocaine esterase (CocE) as a BZE‐metabolizing enzyme (BZEase), which can degrade BZE into biological inactive metabolites (ecgonine and benzoic acid). CocE was redesigned by a reactant‐state‐based enzyme design theory. An encouraging mutant denoted as BZEase2, presented a >400‐fold improved catalytic efficiency against BZE compared with wild‐type (WT) CocE. In vivo, a single dose of BZEase2 (1 mg kg−1, IV) could eliminate nearly all BZE within only two minutes, suggesting the enzyme has the potential for cocaine overdose treatment and BZE elimination in the environment by accelerating BZE clearance. The crystal structure of a designed BZEase was also determined. [ABSTRACT FROM AUTHOR]

Celiac disease (CD), a chronic inflammatory disorder, is triggered by the ingestion of gluten peptide. Wheat gluten contains gliadins and glutenins fractions, where gliadin peptides are the main cause of CD and nonceliac gluten sensitivity development. Keeping a strict gluten-free diet is the only effective treatment for CD. In recent years, lactic acid bacterial and fungal prolyl endopeptidases (PEP) have been proposed as the sources of proteolytic enzymes for the complete elimination of gluten peptides, and have also proved as a potential common therapeutic agent for CD treatment. Considering these indications, a special focus was devoted to AN-PEP-secreted PEP. Herein, we review the proteolytic enzymes produced by microorganisms, especially by the fungal strain, Aspergillus niger (AN), and discuss its beneficial properties against toxic effects of a-gliadin digestion in affected patients. The present review reveals the importance of proteolytic proteases in industrial applications; from an economic perspective, AN-PEP protease is an appropriate choice for making high-quality gluten-free products. [ABSTRACT FROM AUTHOR]

• Pathogenic mechanism of CD related to deaminated gluten binding to HLA-DQ2 and HLA-DQ8 which results in an autoimmune reaction cascade and intestinal inflammation. • Alternative therapeutic approaches could supplement or even supplant dietary therapy. • Alternative treatment focuses on enzymatic degradation of gluten, inhibition of intestinal permeability, modulation of the immune response1, inhibition of the transglutaminase 2 enzyme, blocking antigen presentation, and induction of tolerance. • MSC infusion and nanoparticle-based drug delivery systems are emerging therapeutic strategies for CD. Celiac disease (CD) is a chronic autoimmune enteropathy and multifactorial disease caused by inappropriate immune responses to gluten in the small intestine. Weight loss, anemia, osteoporosis, arthritis, and hepatitis are among the extraintestinal manifestations of active CD. Currently, a strict lifelong gluten-free diet (GFD) is the only safe, effective, and available treatment. Despite the social burden, high expenses, and challenges of following a GFD, 2 to 5 percent of patients do not demonstrate clinical or pathophysiological improvement. Therefore, we need novel and alternative therapeutic approaches for patients. Innovative approaches encompass a broad spectrum of strategies, including enzymatic degradation of gluten, inhibition of intestinal permeability, modulation of the immune response, inhibition of the transglutaminase 2 (TG2) enzyme, blocking antigen presentation by HLA-DQ2/8, and induction of tolerance. Hence, this review is focused on comprehensive therapeutic strategies ranging from dietary approaches to novel methods such as antigen-based immunotherapy, cell and gene therapy, and the usage of nanoparticles for CD treatment. [ABSTRACT FROM AUTHOR]

La maladie de Fabry est due à des mutations du gène GLA à l'origine d'un déficit de l'activité de l'enzyme lysosomale alpha-galactosidase A (α-gal A) et de l'accumulation intra-tissulaire de globotriaosylcéramide. Une nouvelle approche thérapeutique utilisant le chaperon pharmacologique migalastat a été récemment développée. Ce dernier se fixe, de manière spécifique et réversible, sur le site catalytique d'α-gal A mutées, mal conformées, afin d'éviter leur dégradation par le système contrôle-qualité du réticulum endoplasmique et leur permettre de cataboliser le globotriaosylcéramide dans les lysosomes. Ce traitement concerne environ 35 % des mutations du gène GLA reconnues comme sensible au migalastat selon un test pharmacogénétique in vitro. Deux études pivot de phase III, FACETS : migalastat vs. placebo et ATTRACT : migalastat vs. enzymothérapie ont analysé les effets in vivo du migalastat. Malgré certaines limitations méthodologiques, des résultats prometteurs ont été constatés. Le migalastat semble plus efficace que l'enzymothérapie à réduire l'indice de masse du ventricule gauche calculé par échocardiographie en cas d'hypertrophie cardiaque et présente des effets rénaux comparables. Ce traitement oral est le premier traitement personnalisé, axé sur le profil génétique du patient Fabry et ouvre une nouvelle ère dans la prise en charge des maladies conformationnelles. Fabry disease is due to mutations in the GLA gene that cause a deficiency of the activity of the lysosomal enzyme alpha-galactosidase A (α-gal A) resulting in intra-tissue accumulation of globotriaosylceramide. Recently, a novel therapeutic approach based on the pharmacological chaperone migalastat has been developed. It binds, in a specific and reversible manner, to the catalytic site of α-gal A mutants, to prevent their degradation by the quality control system of the endoplasmic reticulum and allow them to catabolize globotriaosylceramide in the lysosomes. This treatment concerns approximately 35% of the GLA gene mutations recognized as sensitive to migalastat according to an in vitro pharmacogenetic test. Two pivotal Phase III studies, FACETS: migalastat vs. placebo and ATTRACT: migalastat vs. enzyme replacement therapy analyzed the in vivo effects of migalastat. Despite some methodological limitations, promising results were found. Migalastat seems to be more effective than enzyme replacement therapy in reducing left ventricular mass index in case of cardiac hypertrophy and has comparable renal effects. This oral treatment is the first personalized treatment, based on the genetic profile of Fabry patients and opens a new era in the management of conformational diseases. [ABSTRACT FROM AUTHOR]

Introduction: Gliadin in flour products is glutamine- and proline-rich and resistant to proteolytic breakdown by gastric and intestinal digestive proteases. Undigested gliadin polypeptides are transformed by tissue transglutaminase into toxic and immunogenic peptides, activating Th1 and Th2 inflammatory pathways. Enzyme therapy, which digests gliadin into non-immunogenic and non-toxic peptides, can be a promising therapeutic approach for celiac disease. Zingibain is a cysteine protease derived from ginger with strong anti-inflammatory properties that can cleave proteins from proline residues. The purpose of this study is to investigate the effect of the zingibain on gliadin digestion. Methods and Materials: At first, the cleavage site of zingibain was simulated on the gliadin sequence. Then, the toxicity and immunogenicity of digested fragments were predicted using ToxinPred and IEDB servers. In the following, the zingibain enzyme was purified from fresh ginger. Gliadin was treated with zingibain at a ratio of 1 to 10 (enzyme: substrate) at 37°C for 16 hours, and the band's pattern of gliadin was investigated by SDS-PAGE 12%. Also, the toxicity of zingibain on the Caco-2 cell line was determined by the MTT method. Results: The results obtained from the simulation of enzymatic cleavage of the gliadin sequence by zingibain and investigating the toxicity and immunogenicity of the resulting fragments with ToxinPred and IEDB servers showed that the zingibain can digest gliadin into non-toxic and non-immunogenic peptides. Also, SDS-PAGE gel demonstrated that zingibain is well able to digest gliadin. Furthermore, the extracted zingibain showed no toxicity on Caco-2 cells up to a concentration of 1000 (μg/mL). Conclusion: Considering the digestion effect of zingibain on gliadin as well as its anti-inflammatory property, by conducting more studies, we can hope for the therapeutic potential of this natural enzyme in the treatment of celiac disease. [ABSTRACT FROM AUTHOR]

4-Chloroisocoumarin compounds have broad inhibitory properties against serine proteases. Here, we show that selected 3-alkoxy-4-chloroisocoumarins preferentially inhibit the activity of the conserved serine protease High-temperature requirement A of Chlamydia trachomatis . The synthesis of a new series of isocoumarin-based scaffolds has been developed, and their anti-chlamydial properties were investigated. The structure of the alkoxy substituent was found to influence the potency of the compounds against High-temperature requirement A, and modifications to the C-7 position of the 3-alkoxy-4-chloroisocoumarin structure attenuate anti-chlamydial properties.

Purpose: Streptococcus pneumoniae (Spn) serotype 3 (Spn3) is considered one of the most virulent serotypes with resistance to conventional vaccine and treatment regimens. Pn3Pase is a glycoside hydrolase that we have previously shown to be highly effective in degrading the capsular polysaccharide of type 3 Spn, sensitizing it to host immune clearance. To begin assessing the value and safety of this enzyme for future clinical studies, we investigated the effects of high doses of Pn3Pase on host cells and immune system. Methods: We assessed the enzyme's catalytic activity following administration in mice, and performed septic infection models to determine if prior administration of the enzyme inhibited repeat treatments of Spn3-challenged mice. We assessed immune populations in mouse tissues following administration of the enzyme, and tested Pn3Pase toxicity on other mammalian cell types in vitro. Results: Repeated administration of the enzyme in vivo does not prevent efficacy of the enzyme in promoting bacterial clearance following bacterial challenge, with insignificant antibody response generated against the enzyme. Immune homeostasis is maintained following high-dose treatment with Pn3Pase, and no cytotoxic effects were observed against mammalian cells. Conclusions: These data indicate that Pn3Pase has potential as a therapy against Spn3. Further development as a drug product could overcome a great hurdle of pneumococcal infections. [ABSTRACT FROM AUTHOR]

Enzyme therapy has important implications for the treatment of metabolic disorders and biological detoxification. It remains challenging to prepare enzymatic nanoreactors with high therapeutic efficiency and low emission of cytotoxic reaction intermediates. Here, we propose a novel strategy for the preparation of enzymes-loaded polypeptide microcapsules (EPM) with concentrically encapsulated enzymes to achieve higher cascade reaction rates and minimal emission of cytotoxic intermediates. Mesoporous silica spheres (MSS) are used as a highly porous matrix to efficiently load a therapeutic enzyme (glucose oxidase, GOx), and a layer-by-layer (LbL) assembly strategy is employed to assemble the scavenging enzyme (catalase) and polyelectrolyte multilayers on the MSS surface. After removal of the MSS, a concentrically encapsulated EPM is obtained with the therapeutic enzyme encapsulated inside the capsule, and the scavenging enzyme immobilized in the polypeptide multilayer shell. Performance of the concentrically encapsulated GOx-catalase capsules is investigated for synergistic glucose metabolism disturbance correction and cytotoxic intermediate H2O2 clearance. The results show that the EPM can simultaneously achieve 99% H2O2 clearance and doubled glucose consumption rate. This strategy can be extended to the preparation of other dual- or multi-enzyme therapeutic nanoreactors, showing great promise in the treatment of metabolic disorders.

The aim of this review is to analyze reasons for the high cost of recombinant human trypsin, technological and economic obstacles limiting trypsin production and implementation, as well as practical means to solve these problems.Materials and methods. The properties of human trypsin, the recombinant technology for its production, the marketing aspects and the prospective of enzyme therapy are addressed in this review that contains 44 references in Russian and english. Particular attention is paid to the methods that can boost the production of recombinant trypsin.Results. Trypsin purified from the mammalian pancreas has been used for over 80 years in the enzyme therapy in various diseases. Genetically engineered human trypsin is proposed to be an innovative, safe and effective therapeutic protease. However the medical use of recombinant trypsin is slowed by its very high price and insufficient production. There is a need for novel biopharmaceutical technologies, as well optimized up-stream and down-stream processes to increase the yield of active recombinant trypsin and significantly reduce the production costs. Recombinant human trypsin that is priced similar to its animal analogues is preferable in all types of enzyme therapy.Conclusion. Innovative biopharmaceutical technologies are expected to significantly reduce the production costs of recombinant human trypsin and stimulate its wider use in enzyme therapy and also in production of other therapeutic proteins.

The literature review considers the problems of the treatment of abdominal pain syndrome, pancreatic exocrine insufficiency syndrome, chronic pancreatitis of Oddi’s sphincter dysfunction, disorders of gall bladder, bile ducts, dysbiosis, intestinal dyskinesia against the background of the latest European recommendations and international con­sensus.

Despite the considerable success in managing the medical process and medical screening of women of childbearing age, the treatment of vaginal infection remains an urgent problem that requires periodic correction of the prescribed therapy. This made the authors conduct a study, which was aimed at investigating the optimization aspects of the comprehensive therapy regimens for vaginal infections (VI) that combine the use of etiotropic drugs with the extract of Aloe Barbadenis Multi-Gyn ActiGel and systemic enzyme therapy as compared to the prescription of monotherapeutic etiotropic regimen. The study evaluated the effectiveness of different treatment regimens of VIs based on the dynamics of clinical and laboratory parameters, as well as the safety and tolerability of different therapies based on the registration of adverse events.Based on the study results, we would advise to include the plant extract of Aloe Barbadenis Multi-Gyn ActiGel and Wobenzym into the therapy of patients with VIs induced by the vaginal dysbiosis and caused by bacterial vaginosis and candidiasis vulvovaginitis-associated microorganisms to improve the elimination of the pathogen and provide a normal titre of lactobacilli with preservation of their functional activity, which eventually restores the impaired vaginal microcenosis and maintains a normal vaginal flora and simultaneously provides a high safety and good tolerability of the proposed treatment regimen.