Development challenges of high concentration monoclonal antibody formulations.

High concentration monoclonal antibody drug products represent a special segment of biopharmaceuticals. In contrast to other monoclonal antibody products, high concentration monoclonal antibodies are injected subcutaneously helping increase patient compliance and reduce the number of hospital patient visits. It is important to note that a high protein concentration (≥50 mg/mL) poses a challenge from a product development perspective. Colloidal properties, physical and chemical protein stability should be considered during formulation, primary packaging and manufacturing process development as well as optimization of other dosage form-related parameters. The aim of such development work is to obtain a drug product capable of maintaining appropriate protein structure throughout its shelf-life and ensure proper and accurate dosage upon administration.

Dataset for landscape pattern analysis from a climatic perspective.

Revealing the driving forces of changes in landscape pattern is a key question of landscape ecology and landscape analysis. Temperature and precipitation as climatic variables have a dominant role in triggering vegetation changes; thus, a database, which contain their interaction, can support the understanding of spatio-temporal changes in vegetation patterns even on a large scale. The dataset provided in this article contain the R-squared values of bivariate linear regression analysis between the Normalized Difference Vegetation Index (target variable; as a general quantitative descriptor of surface greenness) of the TERRA satellite's MODIS sensor and the climatic variables of the CarpatClim database (predictor variables; maximum monthly temperature, aridification index, evapotranspiration and precipitation). Environmental variables are also included to support further analysis: terrain height, macro regions, land cover classes. The dataset has a spatial projection (i.e. maps) and covers the area of Hungary. Tabular version provides the possibility of traditional statistical analysis, while maps allow the investigation to involve the spatial characteristics of absolute and relative position of the data points. This data article is related to the paper "NDVI dynamics as reflected in climatic variables: spatial and temporal trends - a case study of Hungary" (Szabo et al., 2019).

Electroporation-enhanced transdermal diclofenac sodium delivery into the knee joint in a rat model of acute arthritis.

PURPOSE: Since electroporation (EP) can increase the permeability of biological membranes, we hypothesized that it offers an opportunity to enhance the transdermal delivery of drugs for intra-articular indications. Our aim was to compare the anti-inflammatory and analgesic efficacy of EP-combined topical administration of diclofenac sodium hydrogel (50 mg mL-1 in 230 µL volume) with that of an equivalent dose of oral (75 mg kg-1) and simple topical administration. METHODS: Arthritis was induced with the injection of 2% λ-carrageenan and 4% kaolin into the right knee joints of male Sprague Dawley rats. EP was applied for 8 min with 900 V high-voltage pulses for 5 ms followed by a 20 ms break. Drug penetration into the synovial fluid and plasma was detected by high-performance liquid chromatography. Leukocyte-endothelial interactions were visualized by intravital videomicroscopy on the internal surface of the synovium. Inflammation-induced thermal and mechanical hyperalgesia reactions, knee joint edema, and inflammatory enzyme activities were assessed at 24 and 48 h after arthritis induction. RESULTS: EP significantly increased the plasma level of diclofenac as compared with the topical controls 10 min after the 2% λ-carrageenan and 4% kaolin injection. Increased leukocyte-endothelial interactions were accompanied by joint inflammation, which was significantly reduced by oral and EP diclofenac (by 45% and by 30%, respectively) and only slightly ameliorated by simple topical diclofenac treatment (by 18%). The arthritis-related secondary hyperalgesic reactions were significantly ameliorated by oral and EP-enhanced topical diclofenac treatments. The knee cross-section area (which increased by 35%) was also reduced with both approaches. However, simple topical application did not influence the development of joint edema and secondary hyperalgesia. CONCLUSION: The study provides evidence for the first time of the potent anti-inflammatory and analgesic effects of EP-enhanced topical diclofenac during arthritis. The therapeutic benefit provided by EP is comparable with that of oral diclofenac; EP is a useful alternative to conventional routes of administration.

Penetration of lycopsamine from a comfrey ointment through human epidermis.

Mutagenic and teratogenic pyrrolizidine alkaloids (PAs) have been identified in several plant species. The industrially most important PA-containing plant is Symphytum officinale (common comfrey). The application of its root is restricted in several countries due to its PA content. In medicines, the daily alkaloid quantity and duration of treatment may be limited even in case of topical application. Due to the confirmed good absorption of PAs from the gastrointestinal tract, the prohibition of oral use is justified, however the limitation of external application is not supported by relevant data. Penetration experiments on human skin are not available to be a rational basis for limitation. The aim of our work was to carry out pharmacokinetic studies on the diffusion and penetration of lycopsamine (a main PA of comfrey) from a Symphytum product through a synthetic membrane and human skin. Investigations were carried out on vertical Franz diffusion cell and lycopsamine was quantified by a validated LC-MS method. The amount of lycopsamine diffused through a synthetic membrane varied between 0.11% and 0.72% (within 24 h). On human epidermis, the rate of penetration was lower (0.04-0.22%). Our results may contribute to the more realistic toxicological assessment of externally applied PA-containing products.

[Investigating the predictive value of RMI and ROMA indices in patients with ovarian tumors of uncertain dignity]. / Az RMI- és a ROMA-index petefészekrákelõrejelzõ hatékonyságának vizsgálata adnextumorokban.

We were investigating the predictive value of RMI and ROMA indices in patients with ovarian tumors of uncertain dignity, in order to determine whether these methods are suitable for the early detection of ovarian malignancy. Our study included 162 patients treated at the Gynecological Department of the National Institute of Oncology (Budapest, Hungary). These patients were diagnosed with ovarian tumor of uncertain dignity, and were admitted to our Department with the purpose of gynecological surgery. Each of them had CA-125, HE4 blood tests and ultrasound scan in order to calculate RMI and ROMA indices and to study their effectiveness. In every case, the final type of surgery was determined by intraoperative frozen section examination results. Efficacy of RMI and ROMA indices was detected by the final histological examination taken from the same material that was sent for intraoperative frozen section. The sensitivity and specificity of RMI index was 82.0% and 85.1%, respectively, while ROMA index sensitivity and specificity was 88.5% and 72.3%. The results were better in postmenopausal women: RMI sensitivity had increased to 90.9% and specificity to 82.8%. ROMA index sensitivity reached 95.5% with a specificity of 60.7%. Thus premenopausal RMI sensitivity significantly decreased (58.8%), and specificity had surged (88.4%). In case of premenopausal ROMA results sensitivity had declined, though the results are much better than for RMI (70.6% vs. 58.8%), while specificity was 14% less than that of RMI (74.4% vs. 88.4%). According to our study, RMI and ROMA indices are good methods for identifying the dignity of malignant ovarian tumors. The sensitivity and specificity results are in accordance with international literature. Even though the premenopausal and postmenopausal values are different, RMI and ROMA tests complement each other and are excellent for predicting the dignity of a tumor. With the help of these indices 61 cases of malignancy were detected, which means that we have to operate only 3 patients in order to detect 1 case of malignancy.

Electroporation-delivered transdermal neostigmine in rats: equivalent action to intravenous administration.

PURPOSE: Transdermal electroporation has become one of the most promising noninvasive methods for drug administration, with greatly increased transport of macromolecules through the skin. The cecal-contracting effects of repeated transdermal electroporation delivery and intravenous administration of neostigmine were compared in anesthetized rats. METHODS: The cecal contractions were detected with implantable strain gauge sensors, and the plasma levels of neostigmine were followed by high-performance liquid chromatography. RESULTS: Both intravenously and EP-administered neostigmine (0.2-66.7 µg/kg) increased the cecal contractions in a dose-dependent manner. For both the low doses and the highest dose, the neostigmine plasma concentrations were the same after the two modes of administration, while an insignificantly higher level was observed at a dose of 20 µg/kg after intravenous administration as compared with the electroporation route. The contractile responses did not differ significantly after the two administration routes. CONCLUSION: The results suggest that electroporation-delivered neostigmine elicits action equivalent to that observed after intravenous administration as concerning both time and intensity. Electroporation permits the delivery of even lower doses of water-soluble compounds through the skin, which is very promising for clinical practice.

Investigation of the Efficacy of Transdermal Penetration Enhancers Through the Use of Human Skin and a Skin Mimic Artificial Membrane.

The aim of this study was to investigate the behavior of promising penetration enhancers through the use of 2 different skin test systems. Hydrogel-based transdermal formulations were developed with ibuprofen as a nonsteroidal anti-inflammatory drug. Transcutol and sucrose esters were used as biocompatible penetration enhancers. The permeability measurements were performed with ex vivo Franz diffusion cell methods and a newly developed Skin Parallel Artificial Membrane Permeability Assays (PAMPA) model. Franz diffusion measurement is commonly used as a research tool in studies of diffusion through synthetic membranes in vitro or penetration through ex vivo human skin, whereas Skin PAMPA involves recently published artificial membrane-based technology for the fast prediction of skin penetration. It is a 96-well plate-based model with optimized artificial membrane structure containing free fatty acid, cholesterol, and synthetic ceramide analog compounds to mimic the stratum corneum barrier function. Transdermal preparations containing 2.64% of different sucrose esters and/or Transcutol and a constant (5%) of ibuprofen were investigated to determine the effects of these penetration enhancers. The study demonstrated the good correlation of the permeability data obtained through use of human skin membrane and the in vitro Skin PAMPA system. The Skin PAMPA artificial membrane serves as quick and relatively deep tool in the early stages of transdermal delivery systems, through which the enhancing efficacy of excipients can be screened so as to facilitate the choice of effective penetration components.

ATR-FTIR and Raman spectroscopic investigation of the electroporation-mediated transdermal delivery of a nanocarrier system containing an antitumour drug.

The aim of the present work was the optimization of the transdermal delivery of a lyotropic liquid crystal genistein-based formulation (LLC-GEN). LLC was chosen as medium in view of the poor solubility of GEN in water. Membrane diffusion and penetration studies were carried out with a Franz diffusion cell, through a synthetic membrane in vitro, a chick chorioallantoic membrane ex ovo, and ex vivo excised human epidermis. Thereafter, LLC-GEN was combined with electroporation (EP) to enhance the transdermal drug delivery. The synergistic effect of EP was verified by in vivo ATR-FTIR and ex vivo Raman spectroscopy on hairless mouse skin.

The Effect of Electroporation of a Lyotroic Liquid Crystal Genistein-Based Formulation in the Recovery of Murine Melanoma Lesions.

A lamellar lyotropic liquid crystal genistein-based formulation (LLC-Gen) was prepared in order to increase the aqueous solubility of the lipophilic phytocompound genistein. The formulation was applied locally, in a murine model of melanoma, with or without electroporation. The results demonstrated that, when the formulation was applied by electroporation, the tumors appeared later. During the 21 days of the experiment, the LLC-Gen formulation decreased the tumor volume, the amount of melanin and the degree of erythema, but when electroporation was applied, all these parameters indicated a better prognosis even (lower tumor volume, amount of melanin and degree of erythema). Although hematoxylin-eosin (HE) staining confirmed the above events, application of the LLC-Gen formulation by electroporation did not lead to a significant effect in terms of the serum concentrations of the protein S100B and serum neuron specific enolase (NSE), or the tissue expression of the platelet-derived growth factor receptor ß (PDGFRß) antibody.