Diphosphate Choline Research Articles

Advances in the synthesis of cytidine-5'-diphosphate choline

Cytidine-5'-diphosphate choline (CDP-choline) plays a crucial role in the formation of the phospholipid bilamolecular layer in cell membranes and the stabilization of the neurotransmitter system, acting as a precursor to phosphatidylcholine and acetylcholine. CDP-choline has been found effective in treating functional and consciousness disorders resulting from brain injury, Parkinson's disease, depression and glaucoma, and other conditions. As such, CDP-choline is widely utilized in clinical medicine and health care products. The conventional chemical synthesis process of CDP-choline is gradually being replaced by biosynthesis due to the expensive and toxic reagents involved, the production of various by-products, and the high cost of industrial production. Biosynthesis of CDP-choline offers two strategies: microbial fermentation and biocatalysis. Microbial fermentation utilizes inexpensive raw materials but results in a relatively low conversion rate and requires a complex separation and purification process. Biocatalysis, on the other hand, involves two stages: the growth of a living "catalyst" and the conversion of the substrate. Although the synthetic process in biocatalysis is more complex, it offers a higher conversion ratio, and the downstream processing technique for extraction is relatively less costly. Consequently, biocatalysis is currently the primary strategy for the industrial production of CDP-choline. This review aims to summarize the progress made in both chemical synthesis and biosynthesis of CDP-choline, with particular focus on the metabolic pathway and the synthetic processes involved in biocatalysis, in order to provide insights for the industrial production of CDP-choline.

Metabolomics revealed that toxic cyanobacteria stress reduced the flavor quality of Bellamya aeruginosa

The environmental impact of increased eutrophication and frequent cyanobacterial blooms on the growth and reproduction of aquatic organisms has become increasingly prominent in recent years. Bellamya aeruginosa is a nutrient-rich aquatic snail consumed by humans, and environmental changes may decrease its meat quality. However, the specific characteristics of muscle metabolism and flavor quality in freshwater snails and their response patterns to bloom-forming cyanobacterial stress have not been reported. Therefore, in this study, alterations in metabolites in the snail muscle after following feeding on toxic Microcystis aeruginosa were investigated using untargeted metabolomics. A total of 1,128 metabolites were identified, and the metabolic pathways of unsaturated fatty acid biosynthesis and purine, glutathione, and glycerophospholipid metabolism in snails fed toxic cyanobacteria differed from those in snails fed Chlorella vulgaris. Quantitative analysis showed increased levels of bitter-free amino acids, such as tyrosine, phenylalanine, and histidine, after consuming toxic cyanobacteria for 42 d, whereas a relative decrease was observed in the levels of umami- and sweet-free amino acids, such as glutamic acid, aspartic acid, serine, threonine, and glicine. The muscles of snails fed toxic cyanobacteria exhibited higher hypoxanthine and hypoxanthine nucleoside and lower adenosine triphosphate, adenine nucleotide, adenine nucleoside diphosphate, and hypoxanthine nucleotide contents than those of snails fed C. vulgaris. Furthermore, increased metabolites, such as stearic acid, palmitic acid, and cytidine diphosphate choline, and decreased metabolites, such as docosapentaenoic acid, docosahexaenoic acid, adrenoic acid, and L-glutamic acid, reflect the harmful effects of toxic cyanobacteria on the nutritional value and flesh quality of B. aeruginosa. This study comprehensively evaluated the effects of cyanobacterial blooms on freshwater gastropods, providing an important theoretical basis for the quality, safety, and sustainable development of snails as food and related processing industries.

Preventive effects of antenatal CDP-choline in a rat model of neonatal hyperoxia-induced lung injury.

Antenatal steroid administration to pregnant women at risk of prematurity provides pulmonary maturation in infants, while it has limited effects on incidence of bronchopulmonary dysplasia (BPD), the clinical expression of hyperoxia-induced lung injury (HILI). Cytidine-5'-diphosphate choline (CDP-choline) was shown to alleviate HILI when administered to newborn rats. Therefore, we investigated effects of maternal administration of CDP-choline, alone or in combination with betamethasone, on lung maturation in neonatal rats subjected to HILI immediately after birth. Pregnant rats were randomly assigned to one of the four treatments: saline (1 mL/kg), CDP-choline (300 mg/kg), betamethasone (0.4 mg/kg), or CDP-choline plus betamethasone (combination therapy). From postnatal day 1 to 11, pups born to mothers in the same treatment group were pooled and randomly assigned to either normoxia or hyperoxia group. Biochemical an d histopathological effects of CDP-choline on neonatal lung tissue were evaluated. Antenatal CDP-choline treatment increased levels of phosphatidylcholine and total lung phospholipids, decreased apoptosis, and improved alveolarization. The outcomes were further improved with combination therapy compared to the administration of CDP-choline or betamethasone alone. These results demonstrate that antenatal CDP-choline treatment provides benefit in experimental HILI either alone or more intensively when administered along with a steroid, suggesting a possible utility for CDP-choline against BPD.

Choline and citicoline ameliorate oxidative stress in acute kidney injury in rats.

The purpose of this study is to investigate the effects of cholinergic anti-inflammatory pathway (CAP)-activating drugs, choline and citicoline (Cytidinediphosphate-choline, CDP-choline), on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) parameters and the contribution of NADPH Oxidase4 (NOX4) p22phox. Endotoxemia induces a systemic inflammatory response characterized by the production of pro-inflammatory mediators and reactive oxygen species (ROS), which eventually develops acute kidney injury (AKI). NADPH Oxidase4 (NOX4) p22phox pathway contributes to the development of endotoxemia-induced AKI. Inflammatory response can be controlled by CAP. Expressions levels of KIM-1, TNF-α, NOX4, p22phox and NFκB in the kidney tissues of rats were analyzed via RT-PCR in experimental groups; 1. Control, 2. LPS (10 mg/kg) + saline, 3. LPS + CDP-choline (375 mg/kg) and 4. LPS + choline (90 mg/kg). Choline and ROS levels in kidney tissues were also measured by a spectrofluorometric assay. LPS-induced elevations of ROS levels were decreased by CDP-choline or choline administration (p < 0.001). LPS-elevated KIM-1, TNFα, NOX4, p22 phox, and NFκB expressions were significantly decreased by choline or CDP-choline treatments (p < 0.001). Decreased ROS production in kidney tissues in treatment groups suggests that choline or CDP-choline may have therapeutic potential in endotoxemia-associated AKI via downregulating NOX4 and p22phox expressions (Tab. 1, Fig. 5, Ref. 45). Text in PDF www.elis.sk Keywords: endotoxemia, choline, cytidine diphosphate choline, acute kidney injury, reactive oxygen species.

Effects of cytidine-5′-diphosphate choline on gray matter volumes in methamphetamine-dependent patients: A randomized, double-blind, placebo-controlled study

BackgroundCytidine-5′-diphosphate choline (CDP-choline) has been suggested to exert neuroprotective and neuroreparative effects and may be beneficial for patients with stimulant dependence. This randomized, double-blind, placebo-controlled study in methamphetamine (MA) dependence investigated effects of CDP-choline on the brain structures and their associations with craving and MA use. MethodsMA users (n = 44) were randomized to receive 2 g/day of CDP-choline (n = 22) or placebo (n = 22) for 8 weeks. Patients underwent brain magnetic resonance imaging (MRI) at baseline and 8-week follow-up. Healthy individuals (n = 27) were also examined using brain MRI at the same interval. Voxel-based morphometry analysis was conducted to examine changes in gray matter (GM) volumes and their associations with craving and MA use. ResultsCraving for MA was significantly reduced after the 8 week-treatment with CDP-choline (p = 0.01), but not with the placebo treatment (p = 0.10). There was no significant difference in the total number of MA-negative urine samples between the two groups (p = 0.19). With CDP-choline treatment, GM volumes in the left middle frontal gyrus (p = 0.001), right hippocampus (p = 0.009), and left precuneus (p = 0.001) were significantly increased compared to the placebo and control groups. Increased GM volumes in the left middle frontal gyrus with CDP-choline treatment were associated with reduced craving for MA (Spearman's ρ = −0.56, p = 0.03). In addition, the right hippocampal volume increases were positively associated with the total number of MA-negative urine results in the CDP-choline group (Spearman's ρ = 0.67, p = 0.006). ConclusionOur findings suggest that CDP-choline may increase GM volumes of MA-dependent patients, which may be related to decreases in MA use and craving.

A Procedure to Design One-Pot Multi-enzyme System for Industrial CDP-Choline Production.

Fermentation and chemical methods for industrial cytidine diphosphate choline (CDP-choline) catalytic production both suffer from several disadvantages such as relatively low efficiency and productivity. To overcome these problems, we applied the concept of synthetic biology to develop a new one-pot multi-enzyme system to produce CDP-choline from orotic acid. Enzymes from different sources were selected and optimized as building blocks of the system, and parameters such as oxygen supply were also optimized. This system shows a titer of 37.6 ± 1.1 mM and a reaction rate of 1.6 mM L-1 h-1, both increase 66 % from traditional processes. It also has an efficiency of energy of 25.4%, improves 2-folds. This new one-pot CDP-choline-producing system has a potential for industrial use, and the procedure to design one-pot multi-enzyme system can be applied to build other one-pot system producing energy-rich compounds.

Central Injection of Ghrelin Improves Motor Balance in the Rotarod Test in the Rats: Altering the Expression of Drd1 Gene

Introduction: Motor learning consolidates in adulthood, and its defects begin to appear with aging. Ghrelin, an endogenous peptide, improves memory and learning, targeting dopaminergic circuits. While cytidine diphosphate choline (citicoline) is known as a common drug for enhancing memory and learning in aging, it is not recommended for adults due to its side effects. The current study aimed at investigating if ghrelin treatment would improve motor learning via the expression of a relevant gene. Methods: For this experimental study, adult male Wistar rats were randomly divided into five groups: control group, three groups of ghrelin treatment (0.3, 1.5, and 3 nmol/μL), and one group with citicoline treatment. The injections were done intra-hippocampally. The motor learning rate was determined using the rotarod performance test by measuring the resistance to falling. Then the expression of dopamine receptor type D1 (Drd1) gene in the hippocampus was measured by a real-time polymerase chain reaction (PCR). Results: Ghrelin (3 nmol/μL) and citicoline had similar and significant effects on motor learning improvement (P&lt;0.01). Both drugs significantly increased Drd1 gene expression (P &lt;0.001). Conclusion: Ghrelin, like citicoline, improves motor learning by altering the expression of Drd1 gene in the hippocampus.

The clinical effect of citicoline sodium combined with Shenxiong glucose in the treatment of senile hypertensive cerebral infarction in the elders

Objective To investigate the clinical therapeutic effect of citicoline sodium combined with Shenxiong glucose in the treatment of senile hypertensive cerebral infarction in the elders. Methods 80 elderly patients with senile hypertensive cerebral infarctiontreated in our hospital were selected and randomly divided into the single treatment group and the combination treatment group, 40 cases in each group. Both groups received the routine treatment. The single treatment group additionally received Shenxiong glucose injection (100 ml/d), while patients in the combination treatment group additionally received Shenqiong glucose injection combined with citicoline sodium intravenous infusion therapy (0.5 g /d), both groups were treated for 2 weeks. The levels of serum inflammatory factors, the neurological deficit score, the cognitive function score were compared and analyzed before and after treatment between two groups. Results After systemic treatment, the blood pressure and blood lipid levels of two groups were significantly improved, but there was no significant difference between the two groups (P>0.05); The serum levels of interleukin (IL)-6, IL-8 and tumor necrosis factor-α (TNF-α) of the combination treatment group improved more significantly (P<0.05). After treatment, the oxidative stress indexes were significantly improved in the two groups (P<0.05). The content of malondialdehyde (MDA) was decreased, while the superoxide dismutase (SOD) activity and nitric oxide (NO) content were increased significantly (P<0.05); and the improvement degree in the combination treatment group was better than in the single treatment group (P<0.05). The degree of improvement in the Modified Edinburgh-Scandinavia Stroke Scale (MESSS) and Hasegawa Dementia Scale (HDS) scores of the combination treatment group was more significant than those in the single treatment group (P<0.05). The total effective rate of the combination treatment group was 92.5%, which was significantly higher than that of the single group (75.0%) , with statistically significant difference (P<0.05). No obvious adverse reactions happened in two groups during treatment. Conclusions Combination of citicoline sodium and shenxiang glucose on the basis of routine treatment can significantly reduce oxidative stress and inflammation levels, promote the recovery of neurological and cognitive functions, and improve the clinical efficacy and safety. It is worth popularizing and applying in the clinical treatment of senile hypertensive cerebral infarction. Key words: Hypertension; Brain infarction; DANSHENSU; TETRAMETHYLPYRAZINE; Cytidine diphosphate choline; Aged

Computation-aided rational design of a halophilic choline kinase for cytidine diphosphate choline production in high-salt condition

Biocatalysis has become the main approach to produce cytidine diphosphate choline (CDP-choline), which has been applied for treatment of acute craniocerebral injury and consciousness after brain surgery. However, salt accumulates with the production and inhibits enzyme activity, and eventually reduces yield and product accumulation rate. Our work provided a possible solution to this problem by applying a computational designed halophilic choline kinase. The halotolerant CKI (choline kinase) was designed following a unique strategy considering the most variable residue positions on the protein surface among target enzymes from different sources. The basic and neutral surface residues were replaced with acidic ones. This approach was enlightened by features of natural halophilic enzymes. Mutants in the work represented higher catalytic activities and IC50 (inhibit activity by 50%) at high salt concentrations (over 1200 mM). Furthermore, when the mutant was used in fed-batch production, the CDP-choline accumulation rate doubled comparing with process using wild-type CKI at acetate concentration of over 700 mM. The maximum titer was 151 ± 3.2 mM, the productivity was 5.8 ± 0.1 mM·L−1 h−1, and molar yield to CMP and utilization efficiency of energy were 85.3 and 63.5%. The idea of computational design in our work can also be applied to modify other enzymes in industry, and sheds light on alleviating effect of salt accumulation during industrial manufacturing process.

Bioinspired carbon quantum dots for sensitive fluorescent detection of vitamin B12 in cell system

Zwitterion-modification, as a bioinspired strategy, provides greatly promising platforms for biological detection and sensor applications. A green, low-cost and straight-forward method for synthesis of highly fluorescent biomimetic carbon quantum dots (BCQDs) has been developed via pyrolysis of cytidine diphosphate choline (CDPC) and ethylenediamine. The BCQDs with a strong emission at wavelength of 450 nm shows ultrasensitive sensing capability for vitamin B12 with high selectivity. Using the fluorometric assay, the detection limit (DL) for vitamin B12 was found to be as low as 81 nM. Meanwhile, the results of 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT), hemolysis measuring and morphological characterization of Red blood cells (RBCs) confirms the excellent biocompatibility of BCQDs. The imaging experiments of human cervical cancer cells (HeLa) certify that BCQDs could be served as an effective fluorescent sensing probe for label-free sensitive and selective detection of vitamin B12 in biological samples on account of their low toxicity and good biocompatibility. The BCQDs, further, were successfully applied to probe vitamin B12 in living cells, which broaden its potential application in vivo system.

Rational Design of an Efficient Halotolerant Enzymatic System for In Vitro One-Pot Synthesis of Cytidine Diphosphate Choline.

Salt accumulation often impedes cytidine diphosphate choline (CDP-choline) in vitro biosynthetic process. In this work a halotolerant in vitro enzymatic system is developed to solve this problem. It applies a halotolerant choline-phosphate cytidylyltransferase (CCT) obtained from rational design instructed by a unique strategy, which refers to one of the features of naturally occurring halophilic enzymes. By increasing acidic residues on protein surface where is most variable with respect to amino acid in the sequence alignment with other CCT, the mutants are obtained. The mutants represent higher catalytic activities and IC50 values (inhibit activity by 50%) at high-salt concentrations. Furthermore, when the halotolerant CCT is applied to in vitro one-pot biosynthesis of CDP-choline, the maximum titer and productivity are 161 ± 3.5 mM and 6.2 ± 0.1 mM L-1 h-1 , respectively. When acetate concentration increases, it still keeps relatively high reaction rate and is 2.2-fold higher than process using wild-type CCT (3.87 mM L-1 h-1 comparing with 1.74 mM L-1 h-1 ). This halotolerant system has great potential for industrial use, and the rational design concept can be applied to modify other enzymes, addressing the salt accumulation problem in in vitro systems, and gives insight into resolving by-product inhibition during reaction.

Citicoline in Vascular Cognitive Impairment: Some Latest Evidences

CDP-choline (cytidine-5′-diphosphate choline), also called citicoline, is one of the most frequently pre­scribed drugs for cognitive impairment in several European countries and worldwide [1–3].

Clinical medication characteristics of Shuxuening injection in treatment of cerebral infarction research based on registration

To understand the medication characteristics of Xingxue@Shuxuening solutions in the real world, multi-center, large-sample-size registration design method is adopted in this study. Between October 2012 and October 2015, hospitalized patients in 27 medical institutions who used Shuxuening and diagnosed as cerebral infarction were observed, including their general information, diagnosis information and medication information. Totally 9 473 cases of cerebral infarction were included, with old people as the majority, and males were slightly more men than females; they were complicated with coronary heart disease, vertebrobasilar artery insufficiency, transient brain ischemia and other cerebrovascular diseases; the medication duration was mostly within 14 days; the dosage was mainly 8-20 mL; 0.9%NS, 5%GS were the main solvent; commonly used drug combination was Ozagrel sodium injection, Shuxuetong injection, Alprostadil injection and other medicines for inhibiting platelet aggregation and blood vessel dilatation, and Cytidine diphosphate choline injection, Edaravone injection, Oxiracetam injection and other brain protective agents. The above results indicate that the clinical medication characteristics of Shuxuening are basically consistent with the package insert, except for the solvent; combined medicine is dominated by antiplatelet drugs; for old patients, various anticoagulant drugs shall selected according to their specific conditions to avoid the occurrence of adverse reactions.

Efficient multi-enzyme-catalyzed CDP-choline production driven by an ATP donor module.

Cytidine diphosphate choline (CDP-choline) has been applied for treating acute craniocerebral injury and allowing recovery of consciousness after brain surgery. In this study, an acetate kinase (ACK)/acetyl phosphate system was used to supply ATP and combined with Escherichia coli-overexpressed CMP kinase (CMK), NDP kinase (NDK), choline phosphate cytidylyltransferase (CCT), and choline kinase (CKI) to produce CDP-choline from CMP and choline chloride. Within 1h, 49mM CDP-choline was produced, for a molar yield of 89.9 and 68.4% based on CMP and choline chloride, respectively; the utilization efficiency of energy (UEE) was 79.5%. Acetyl phosphate, sodium acetate, and CTP inhibited the reaction when the concentration exceeded 18.5, 600, and 30mM, respectively. This inhibition could be overcome by controlling the rate of acetyl phosphate, CMP addition or using KOH instead of NaOH to regulate the pH in fed-batch transformation. After 24h, the maximum titer was 124.1±2.7mM, the productivity was 5.1±0.1mMl-1h-1, the molar yield to CMP and choline chloride were 83.8 and 63.7%, respectively, and the UEE was 58.2%. This high yield and productivity of CDP-choline through biocatalysis suggest future application at the industrial scale.

Improved Synthesis of Cytidine Diphosphate Choline (CDP-Choline) Via Selective Phosphorylation

An improved, three-step synthesis of cytidine diphosphate choline (CDP-choline) from cytidine was achieved in 68% overall yield. Selective phosphorylation of cytidine was accomplished by the use of morpholinophosphodichloridate to give cytidine-5′-phosphomorpholide, which was condensed with choline phosphate chloride in the presence of a catalytic amount of H2SO4 to give CDP-choline. The intermediates and products could be efficiently purified by recrystallisation, thus avoiding the use of chromatography at all stages. The reaction could be scaled up to 200 g in 64% overall yield, making this route attractive for industrial application.

1603957: Improved synthesis of cytidine diphosphate choline (CDP-choline) via selective phosphorylation – ESI

1603957: Improved synthesis of cytidine diphosphate choline (CDP-choline) <i>via</i> selective phosphorylation – ESI

Atlantic cod (Gadus morhua) larvae can biosynthesis phospholipid de novo from 2-oleoyl-glycerol and glycerol precursors.

The dietary requirement of phospholipid (PL) of fish larvae has been suggested to originate in an inefficient ability for de novo biosynthesis of PL based on dietary triacylglycerol (TAG). The main objective of the present study was to investigate whether cod larvae could synthesis PL from sn-2-monoacylglycerol (2-MAG) and glycerol precursors. A tube feeding method was used to deliver equal molar aliquots of 2-oleoyl-[1,2,3-(3)H]glycerol and [U-(14)C] glycerol together with bovine serum albumin (BSA) bound 16:0 (palmitic acid) and 22:6n-3 (docosahexaenoic acid, DHA), with or without choline chloride to the foregut of anesthetized cod larvae and thereafter monitoring the metabolism of these components in the larvae through 4 h following injection. Our results showed that both 2-MAG and glycerol precursors contributed to the de novo synthesis of phosphatidylcholine (PC) and the 2-MAG pathway predominated over the G-3-P (glycerol-3-phosphate) pathway in the synthesis of TAG and PC. The molecular ratio of PC/TAG obtained from the 2-MAG and the G-3-P pathways was 0.44-0.74 and 1.02-2.06 within the first hour of tube feeding, suggesting they might have comparable biosynthesis ability of PC and TAG under the conditions of the present study. Furthermore, supplementation of choline chloride significantly increased PC/TAG ratio (p < 0.05) for both pathways. However, further studies are needed to quantify the enzyme activity involved in the CDP-choline (cytidine diphosphate choline) pathway, and the function of choline either in simulating PC synthesis or TAG catabolism or both needs further investigation.

Efficient synthesis of cytidine diphosphate choline (CDP-choline) and its analogs

GRAPHICAL ABSTRACT

Gemcitabine diphosphate choline is a major metabolite linked to the Kennedy pathway in pancreatic cancer models in vivo.

Background:The modest benefits of gemcitabine (dFdC) therapy in patients with pancreatic ductal adenocarcinoma (PDAC) are well documented, with drug delivery and metabolic lability cited as important contributing factors. We have used a mouse model of PDAC: KRASG12D; p53R172H; pdx-Cre (KPC) that recapitulates the human disease to study dFdC intra-tumoural metabolism.Methods:LC-MS/MS and NMR were used to measure drug and physiological analytes. Cytotoxicity was assessed by the Sulphorhodamine B assay.Results:In KPC tumour tissue, we identified a new, Kennedy pathway-linked dFdC metabolite (gemcitabine diphosphate choline (GdPC)) present at equimolar amounts to its precursor, the accepted active metabolite gemcitabine triphosphate (dFdCTP). Utilising additional subcutaneous PDAC tumour models, we demonstrated an inverse correlation between GdPC/dFdCTP ratios and cytidine triphosphate (CTP). In tumour homogenates in vitro, CTP inhibited GdPC formation from dFdCTP, indicating competition between CTP and dFdCTP for CTP:phosphocholine cytidylyltransferase (CCT). As the structure of GdPC precludes entry into cells, potential cytotoxicity was assessed by stimulating CCT activity using linoleate in KPC cells in vitro, leading to increased GdPC concentration and synergistic growth inhibition after dFdC addition.Conclusions:GdPC is an important element of the intra-tumoural dFdC metabolic pathway in vivo.

Diversity and function of membrane glycerophospholipids generated by the remodeling pathway in mammalian cells

Cellular membranes are composed of numerous kinds of glycerophospholipids with different combinations of polar heads at the sn-3 position and acyl moieties at the sn-1 and sn-2 positions, respectively. The glycerophospholipid compositions of different cell types, organelles, and inner/outer plasma membrane leaflets are quite diverse. The acyl moieties of glycerophospholipids synthesized in the de novo pathway are subsequently remodeled by the action of phospholipases and lysophospholipid acyltransferases. This remodeling cycle contributes to the generation of membrane glycerophospholipid diversity and the production of lipid mediators such as fatty acid derivatives and lysophospholipids. Furthermore, specific glycerophospholipid transporters are also important to organize a unique glycerophospholipid composition in each organelle. Recent progress in this field contributes to understanding how and why membrane glycerophospholipid diversity is organized and maintained.