Broad-Spectrum In Vitro Antiviral Activity of ODBG-P-RVn: An Orally-Available, Lipid-Modified Monophosphate Prodrug of Remdesivir Parent Nucleoside (GS-441524).

The necessity for intravenous administration of remdesivir confines its utility for treatment of coronavirus disease 2019 (COVID-19) to hospitalized patients. We evaluated the broad-spectrum antiviral activity of ODBG-P-RVn, an orally available, lipid-modified monophosphate prodrug of the remdesivir parent nucleoside (GS-441524), against viruses that cause diseases of human public health concern, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). ODBG-P-RVn showed 20-fold greater antiviral activity than GS-441524 and had activity nearly equivalent to that of remdesivir in primary-like human small airway epithelial cells. Our results warrant in vivo efficacy evaluation of ODBG-P-RVn. IMPORTANCE While remdesivir remains one of the few drugs approved by the FDA to treat coronavirus disease 2019 (COVID-19), its intravenous route of administration limits its use to hospital settings. Optimizing the stability and absorption of remdesivir may lead to a more accessible and clinically potent therapeutic. Here, we describe an orally available lipid-modified version of remdesivir with activity nearly equivalent to that of remdesivir against emerging viruses that cause significant disease, including Ebola and Nipah viruses. Our work highlights the importance of such modifications to optimize drug delivery to relevant and appropriate human tissues that are most affected by such diseases.

Synergistic Effect and Antibiofilm Activity of a Skin and Wound Cleanser.

INTRODUCTION: Biofilm in chronic wounds impedes the wound healing process. Each biofilm has differing characteristics requiring a multifaceted approach for removal while maintaining a surrounding environment conducive to wound healing. OBJECTIVE: In this study, 3 of the components in a wound cleanser are tested to determine synergy in eradicating biofilms of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa in vitro. MATERIALS AND METHODS: The 3 components assessed for synergy were ethylenediamine tetraacetic acid sodium salts (EDTA), vicinal diols (VD; ethylhexylglycerin and octane-1,2-diol), and polyhexamethylene biguanide (PHMB). Each component was assessed individually and in combination while dissolved in a base solution. The Calgary assay method was used for biofilm growth and treatment. Kull Equation analysis for synergy was conducted using viable count results. RESULTS: Synergy is defined as the interaction of components to produce a combined effect greater than the sum of their separate effects. The base solution containing all 3 components (EDTA, VD, and PHMB) reduced biofilm viability by more than 5 logs, demonstrating statistically significant synergy. The 3 components tested individually in the base solution resulted in the following: EDTA did not reduce bacteria viability; VD reduced viability by about 1 log; and PHMB reduced P aeruginosa viability by about 2.5 logs and MRSA viability by about 4 logs. Of importance, the MRSA biofilm failed to regrow in the recovery plates after combined treatment, indicating complete elimination of the biofilm bacteria. CONCLUSIONS: The experimental and calculated results indicate the 3 components (VD, EDTA, and PHMB) when used together act synergistically to eradicate MRSA and P aeruginosa biofilms in vitro.

Neuroinflammation and adult hippocampal neurogenesis in neuropathic pain and alkyl glycerol ethers treatment in aged mice.

Neuropathic pain is a condition characterized by unpleasant sensory and emotional experiences associated with a number of diseases or injuries affecting the sensory system through various mechanisms. In this study, we focused on the impact of chronic neuropathic pain on the microglial state and hippocampal neurogenesis in aged mice. In addition, we examined the effects of alkyl glycerol ethers (AGE) treatment on behavioral parameters, hippocampal neuronal and microglial plasticity in aged C57BL/6 mice with neuropathic pain. For the induction of neuropathic pain, we used the model of chronic constriction injury (CCI) of the sciatic nerve. We observed painful behavior in animals subjected to CCI, expressed as a decrease in locomotor activity and the development of cold allodynia. A violation of working and long­term memory was also observed. AGE administration reduced the severity of cold allodynia and prevented memory impairment. In addition to behavioral changes, neuropathic pain was accompanied by microglial activation, changes in the hippocampal production of pro­ and anti­inflammatory cytokines, as well as a decrease in neurogenesis. The administration of AGE prevented the neuropathic pain­derived effects, including M1 microglial activation and neurogenesis disruption. However, in vitro experiments demonstrated the pro­inflammatory activation of microglial cells, emphasizing the complexity of the mechanisms underlying the pharmacological effects of AGE. On the whole, the findings of this study demonstrate that AGE treatment prevented behavioral effects of neuropathic pain in mice, and AGE may thus have potential for use in the prevention or treatment of neuropathic pain cognitive and emotional effects. However, as the mechanisms underlying this type of pain are complex, further studies are required to determine the detailed pharmacological effects of AGE.

Adult hippocampal neurogenesis in neuropathic pain and alkyl glycerol ethers treatment.

Neuropathic pain manifested by a number of sensory symptoms is often accompanied by disorders of higher nervous activity, such as memory impairment, depression, anxiety, anhedonia, etc. This emphasizes the involvement of supraspinal structures including the hippocampus in neuropathic pain pathogenesis. In the present study, we focused on the impact of chronic neuropathic pain on hippocampal neurogenesis and microglial state. In addition, we test the effect of alkyl glycerol ethers on hippocampal neuronal and microglial plasticity as well as behavioral parameters. Neuropathic pain was induced using the model of sciatic nerve chronic constriction injury. We found an impairment of working memory and locomotor activity in animals with neuropathic pain, which was prevented by alkyl glycerol ethers treatment. Sciatic nerve ligation in mice contributed to the decrease in hippocampal neurogenesis intensity. Alkyl glycerol ethers administration significantly reduced this effect. Neuropathic pain-associated neurogenesis reduction was accompanied by an increased percentage of Iba1-labeled area in the CA1 hippocampal region on the 14th and 28th days after surgery. In addition, we observed a decrease in hippocampal pro-inflammatory microglia marker CD86 immunostaining on day 28 after surgery in alkyl glycerol ethers-treated mice with sciatic nerve ligation. These results are consistent with data on pro- and anti-inflammatory cytokines expression in the hippocampus. Alkyl glycerol ethers administration increased IL-10 and decreased IL-1ß hippocampal expression in animals with neuropathic pain. Taken together, these data suggest that neuropathic pain-behavior in rodents is accompanied by changes in microglia polarization, thereby contributing to neurogenesis impairment and cognitive disturbances. Alkyl glycerol ethers prevented M1 microglial activation, contributing to the maintenance of normal neurogenesis levels within the hippocampus and normalizing working memory.

Functions of plasmalogen lipids in health and disease.

Plasmalogens are a unique class of membrane glycerophospholipids containing a fatty alcohol with a vinyl-ether bond at the sn-1 position, and enriched in polyunsaturated fatty acids at the sn-2 position of the glycerol backbone. These two features provide novel properties to these compounds. Although plasmalogens represent up to 20% of the total phospholipid mass in humans their physiological roles have been challenging to identify, and are likely to be particular to different tissues, metabolic processes and developmental stages. Their biosynthesis starts in peroxisomes, and defects at these steps cause the malformation syndrome, Rhizomelic Chondrodysplasia Punctata (RCDP). The RCDP phenotype predicts developmental roles for plasmalogens in bone, brain, lens, lung, kidney and heart. Recent studies have revealed secondary plasmalogen deficiencies associated with more common disorders and allow us to tease out additional pathways dependent on plasmalogen functions. In this review, we present current knowledge of plasmalogen biology in health and disease.

An update on the therapeutic role of alkylglycerols.

Scandinavian folk medicine used shark liver oil for the treatment of cancers and other ailments based on the rarity of tumors in sharks and their ability to resist infections. Shark liver oil is a source of alkylglycerols which have been studied as anti-cancer agents in several clinical trials. Moreover, alkylglycerols have been investigated for the treatment of radiation induced side effects and for their ability to boost the immune system. Several experimental studies have shown the ability of alkylglycerols to open the blood brain barrier to facilitate the access of therapeutic drugs to the central nervous system. This review covers the most important studies of alkylglycerols in both animals and humans.

A Pex7 hypomorphic mouse model for plasmalogen deficiency affecting the lens and skeleton.

Rhizomelic chondrodysplasia punctata type 1 is a peroxisome biogenesis disorder with the clinical features of rhizomelia, abnormal epiphyseal calcifications, congenital cataracts, and profound growth and developmental delays. It is a rare autosomal recessive disorder, caused by defects in the peroxisome receptor, PEX7. The pathology results from a deficiency of plasmalogens, a critical class of ether phospholipids whose functions are largely unknown. To study plasmalogens in an animal model, avoid early mortality and facilitate therapeutic investigations in this disease, we engineered a hypomorphic mouse model in which Pex7 transcript levels are reduced to less than 5% of wild type. These mice are born in expected ratios, are fertile and have a normal life span. However, they are petite and develop early cataracts. Further investigations showed delayed endochondral ossification and abnormalities in lens fibers. The biochemical features of reduced Pex7 function were reproduced in this model, including tissue plasmalogen deficiency, phytanic acid accumulation, reduced import of Pex7 ligands and consequent defects in plasmalogen biosynthesis and phytanic acid oxidation. Dietary supplementation with batyl alcohol, a plasmalogen precursor, recovered ether phospholipids in blood, but did not alter the clinical phenotype. The relatively mild phenotype of these mice mimics patients with milder PEX7 defects, and highlights the skeleton and lens as sensitive markers of plasmalogen deficiency. The role of plasmalogens in the normal function of these tissues at various ages can now be studied and additional therapeutic interventions tested in this model.

[Effect of 1-O-alkyl-glyceride ethers isolated from lipids of the squid Berrytteuthis magister liver on lipid metabolism and hematological parameters of rats with experimental dislipidemia].

On the white Wistar rats with alimentary dyslipidemia investigated influence 1-O-alkyl-glycerides ethers (AGE), received by a method of hydrolysis 1-O-alkyl-diacylglycerides from lipids of the squid Berryteuthis magister liver, on a lipid metabolism, hepatobiliary functions of liver, antioxidant systems and parameters of blood. Are revealed antioxidant, antianemia and immunoactive properties of AGE. AGE raise a level of glucose and activity of enzymes hepatobiliary systems in blood, interfere the decrease of a cholesterol in blood.

[Experience with glyciphonic ointment therapy for primary multiple basal cell carcinoma of the skin]. / Opyt lecheniia pervichno-mnozhestvennogo bazal'nokletochnogo raka kozhi glitsifonovoi maz'iu.

The effectiveness of glycifone ointment treatment for primary multiple basal cell carcinoma of the skin was evaluated in 28 patients. 27 cases were cured following one or two courses the duration of which was determined by the number of foci, pattern of tumor and its total surface area.

[Experience with glyciphonic ointment for malignant and precancerous lesions of the skin]. / Opyt primeneniia glitsifonovoi mazi pri rake i predrakovykh zabolevaniiakh kozhi.

Skin was smeared with glyciphonic ointment containing 30% of methylphosphonic diglycidyl ether (Tatkhimfarmpreparaty Company) in 495 patients with histologically confirmed basalioma and 36 patients with senile keratosis. During daily application necrotic tissue was removed. Considerable decomposition of tumor occurred on day 6 or later. It took the wound 9-12, seldom, 15 days to heal, with scars forming on days 15-20. Stable cure was registered (5-7 yrs) in 492 patients with skin basalioma and all cases of senile keratosis. No changes in peripheral blood count or any skin-resorption side-effects were recorded. Several patients suffered hyperemia, edema, itching or local pain.

[Glyciphonic ointment in the treatment of radiation-induced basal cell carcinoma of the skin]. / Lechenie glitsifonovoi maz'iu bazal'nokletochnogo raka kozhi, indutsirovannogo radiatsionnym izlucheniem.

Skin basalcell carcinoma of the head and neck occurring within 1.5-23 yrs on the site of radiation fibrosis were smeared daily with glyciphonic ointment without dressing the wound. Tumor disintegrated after 2-6 applications followed by edema tumor tissue development on days 22-24. After a 2-3 week interval, perifocal edema subsided, and tumor deed shrank leaving a small scab. Basalioma cells were detected underneath. Complete cure was recorded after another 2-3 week therapy in all cases. Only one patient relapsed three years later.

Myocardial salvage by 1-O-hexadecyl-Sn-glycerol: possible role of peroxisomal dysfunction in ischemia reperfusion injury.

A recent study demonstrated biochemical and structural alterations of peroxisomes in rat kidney after ischemia/reperfusion. We examined whether peroxisomes play any role in the pathophysiology of myocardial ischemia/reperfusion injury. Isolated perfused rat heart was made ischemic for 30 min by terminating coronary flow (CF), followed by 30-min reperfusion. Experiments were divided into two groups; the experimental group received 1-O-hexadecyl-Sn-glycerol (chimyl alcohol) (25, 50, and 100 microM) before ischemia, and the control group received an equivalent amount of saline. Two of the experimental groups (50 and 100 microM) demonstrated improved postischemic myocardial performance, as demonstrated by accelerated recovery in left ventricular developed pressure (LVDP) and CF, as well as reduction in the incidence of ventricular fibrillation (VF). However, because the heart rate (HR) was significantly reduced in the 100-microM chimyl alcohol group, subsequent studies were performed with 50 microM chimyl alcohol as the optimal dose. Chimyl alcohol (50 microM) also reduced cellular injury, as evidenced by reduced creatine kinase (CK) release, and decreased development of oxidative stress, as evidenced by reduced formation of malonaldehyde (MDA). Peroxisomal catalase activity was decreased in the control group after ischemia/reperfusion, and chimyl alcohol treatment restored the activity of the enzyme. Our results indicate that chimyl alcohol, a precursor of ether-linked phosphoglyceride biosynthesis, can reduce myocardial ischemia/reperfusion injury, possibly by restoring catalase activity and reducing oxidative stress through synthesis of ether lipids, suggesting a possible role of peroxisomal disorder in ischemia/reperfusion injury.

Aplicaçäo do triglicerídeos de cadeia média (TCM) na prevençäo de úlceras de decúbito / The using of MCT in the ulcer prevent

Aproximadamente 75.000 das 1.000.000 pessoas hospitalizadas a cada ano desenvolvem Ulceras de Decúbito. Graças à tecnologia avançada, hoje temos várias opçöes de tratamento destas úlceras, ainda que isto seja dispendioso para os pacientes e hospitais. Porém, melhor que desenvolver um processo curativo, é poder desenvolver um processo preventivo eficáz. Com o objetivo de avaliar a eficácia do uso tópico de triglicerídeos de cadeia média (TCM) na prevençäo de Ulceras de Decúbito, a autora utilizou o método duplo-cego em 86 pacientes, onde 50 por cento foi submetido ao uso de soluçäo com TCM e 50 por cento ao uso de Soluçäo Glicerinada. Em ambos os casos, houve diminuiçäo da instalaçäo de Ulceras de Decúbito porém, dos pacientes que se submeteram ao uso da soluçäo com TCM, apenas 2 (dois) desenvolveram Ulcera de Decúbito (Grau 1); e dos pacientes que se submeteram ao uso de Soluçäo com Glicerina, 12 (doze) desenvolveram Ulcera de Decúbito (Graus 1 e 2). Concluiu-se que o TCM é de grande auxílio na prevençäo de Ulceras de Decúbito, tem excelente absorçäo para uso tópico, forma uma película protetora à pele, previne escoriaçöes devido à sua alta capacidade de hidrataçäo e proporciona nutriçäo celular local.

[Shark liver oil (alkoxyglycerol) and cancer treatment]. / Hajleverolie (alkoxyglycerol) og cancerbehandling.

Alkoxyglycerol derived from shark liver oil is marketed in Denmark and mentioned in popular articles as a supplementary agent in the treatment of cancer. A questionnaire investigation carried out in the Department of Oncology and Haematology in Odense Hospital revealed that approximately 1/3 of the patients in active neoplastic therapy employed shark liver oil preparations. The clinical investigations of alkoxyglycerol were all carried out on patients with cancer of the uterine cervix. All of the investigations were carried out by the same Swedish research group. Only a minority of the experimental material was blinded. No documentation was found for inhibited tumour growth or reduced mortality resulting from treatment with alkoxyglycerol. The number of cases of irradiation damage were found to be fewer in the groups treated with alkoxyglycerol, but the difference may be partially explained by different subdivision into stages. Alkoxyglycerol results in increase in the leukocyte and thrombocyte counts while higher or lower doses have, apparently, the opposite effect. The available literature concerning the clinical effect of alkoxyglycerol is limited and unsystematic and does not support the employment of alkocyglycerol in the treatment of cancer.

Synthesis and antineoplastic properties of ether-linked thioglycolipids.

Ether-linked glycero-alpha- and beta-D-glucopyranosides and glycero-1-thio-alpha- and beta-D-glucopyranosides have been synthesized by modifications of the Königs-Knorr procedure, and their antitumor activities have been evaluated. The bioactivities of these compounds have been evaluated in five different cell lines (WEHI 3B, C653, X63/OMIL3, R6X-B15, and HL-60) and compared with the activities of 1-O-hexadecyl-2-O-methyl-sn-3-glycerophosphocholine (GPC) and its enantiomer, 3-O-hexadecyl-2-O-methyl-sn-1-GPC. The results indicate that a alpha-D-thioglucopyranoside [1-O-hexadecyl-2-O-methyl-3-S-(alpha-D-1'- thioglucopyranosyl-sn-glycerol)] is selective with respect to its action on target cells, with high activity for killing of WEHI 3B and C653 cells as determined by inhibition of [3H]thymidine incorporation into DNA and HL-60 cell cytotoxicity, but unable to induce aggregation of rabbit platelets at 10(-5) M. The corresponding beta-linked thioglycolipid was ineffective with respect to cytotoxicity against each cell line tested, indicating the importance of configuration at the anomeric position; the beta-thioglycoside was also ineffective with respect to inducing platelet aggregation. 1-O-Hexadecyl-2-O-methyl-sn-3-GPC and 3-O-hexadecyl-2-O-methyl-sn-1-GPC were potent inhibitors of growth of each cell line tested but also caused rabbit platelet aggregation at concentrations greater than or equal to 10(-7) M. Thus, 3-S-(alpha-thioglycopyranosyl)-sn- glycerols bearing a long-chain O-alkyl group at the sn-1 position and a methoxy group at the sn-2 position of glycerol appear to be a promising class of antineoplastic agents with lower risk of inducing thrombosis than the widely studied platelet activating factor analogue, 1-O-octadecyl-2-O-methyl-rac-3-GPC.

In vitro and in vivo cytotoxicity of alkyl lysophospholipid ET-18-OCH3 and thioether lipid BM 41.440.

Screening for cytotoxicity in the clonogenic assay in human tumor xenografts and L1210 mouse leukemia revealed comparable dose-dependent effects of the alkyl lysophospholipid ET-18-OCH3 and the thioether lipid BM 41.440. The efficacy in human tumors only was marginal at low doses. In vivo tests of both agents were carried out in nude mice bearing two of the tumors that proved most sensitive in vitro and in mice inoculated with L1210 leukemia. Only small effects on the growth of the human tumors and no effects on L1210 leukemia were observed. In view of clinical rules for definition of remission, no convincing antitumor effects were obtained.

Biosynthesis and biotransformation of ether lipids.

Some naturally occurring as well as synthetic ether lipids are biologically active. In certain cases, the effects of these substances are enhanced, in others, they are inhibited by compounds that were isolated from natural sources or prepared by chemical synthesis. The biotransformation of natural or "unnatural" ether lipids in microorganisms, plant or animal tissue also can lead to substances that elicit biological effects. The production of such compounds through various biotechnological techniques is a field wide open for future exploration. In addition to animal cell cultures, plant cell cultures may become useful tools in biomedical studies concerned with ether lipids.

Clinical phase I pilot study of the alkyl lysophospholipid derivative ET-18-OCH3.

Sixteen patients suffering from widespread malignant disease, the majority pretreated and found in poor general health, were treated in a phase I pilot study with the alkyl lysophospholipid derivative 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH3). Eleven patients were treated intravenously, and five were given oral therapy. Prolonged IV administration of 15-20 mg/kg/day at a concentration of 5 mg ET-18-OCH3 per 1 ml 20% human serum albumin could be continued safely. The maximum-tolerated dose was either 50 mg/kg as a single injection or 20 mg/kg during daily dispensions. Grade 2-4 toxicity, as pulmonary edema and impairment of hepatic function, then occurred during daily treatment. Toxicity was reversible. Mitogen stimulation and mixed lymphocyte culture studies revealed possible immunosuppressive effects of higher doses of ET-18-OCH3. There were no chromosomal changes in cytogenetic studies. Frequent post-mortem examinations revealed no further toxicity. IV and oral treatment showed few encouraging response data since there were two partial remissions in non-small cell lung cancers and a reduction of leukemic blasts to less than 10% in an acute myelomonocytic leukemia.

Ether lipids and analogs in experimental cancer therapy. A brief review of the Munich experience.

This review covers the work of our laboratory on the antineoplastic activity of some ether lipids and derivatives that are related to platelet-activating factor (PAF). Various 1-O-alkyl lysophospholipid derivatives (ALP) show therapeutic activity in mouse transplant tumor models and in metastatic 3-Lewis lung carcinoma in vivo. However, certain autochthonous mouse leukemias and radiation-induced lymphomas are resistant to ALP treatment. The therapeutic effects of these compounds are partially due to the activation of cytotoxic macrophages and direct cytotoxicity. Approximately 20 ether lipids and derivatives were tested for direct cytotoxicity in cells from human solid tumors and leukemias using [3H]thymidine uptake, trypan blue dye exclusion, human tumor clonogenic assays (HTCA) and cell morphology as criteria. Certain ALP, thioether lysophospholipid-derivatives (TLP), ether-linked lipoidal amines, sn-2 analogs of PAF, and conjugates of ether lipids and cytosine arabinoside were found cytotoxic in a dose- and time-dependent fashion. Cytotoxicity of some of the ether lipids tested is based on destruction of cell membranes. Structure-activity studies were performed to better understand the mechanisms leading to accumulation and cytotoxicity of ALP. Comparative studies with normal bone marrow cells and leukemic blasts from humans revealed preferential anti-leukemic cytotoxicity of three ether lipids.