Ethers of phosphatidylcholine and phosphatidylethanolamine, their lyso forms and mixtures thereof for use in the treatment of diseases or disorders caused by plasmalogen deficiency

Abstract

The present disclosure relates to certain compounds that can be ingested by a human subject to regulate plasmalogen levels and improve health outcomes in the subject. The present disclosure also relates to formulations of the compounds, compositions comprising at least one compound, and uses thereof.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 478,289, filed January 3, 2023, and U.S. Provisional Patent Application No. 63 / 589,539, filed October 11, 2023, the entire contents of each of which are incorporated herein by reference.

[0002] The present disclosure relates to certain precursor compounds, such as lysoalkylphosphatidylcholine (LPC(O)), that can be ingested by a human subject to increase plasmalogen levels and result in improved health outcomes. More specifically, the present disclosure relates to formulations of the precursor compounds to provide optimal elevations in circulating and tissue plasmalogens while maintaining an optimal composition of plasmalogen species. [Background technology]

[0003] Plasmalogens, as defined herein, include a group of plasmenyl-phospholipids, which are major cell membrane components. As part of a broader class of phospholipids, plasmanyl-phospholipids and / or plasmenyl-phospholipids are a unique class of ether phospholipids that are major components of cell membranes. While their biophysical role in cell membranes has been studied, knowledge of their biological role is an important area of ​​new research. Plasmalogens exist primarily as alkenylphosphatidylcholine (PC) and alkenylphosphatidylethanolamine (PE) species, also known as PC(P) and PE(P), respectively. These are characterized by a cis-vinyl ether bond connecting an alkyl chain to the sn-1 carbon position of the glycerol backbone. They also optionally contain an acyl-linked fatty acid ester bond at the sn-2 carbon position. Although plasmalogens are often esterified with polyunsaturated fatty acids such as arachidonic acid (20:4) and the omega-3 fatty acid docosahexaenoic acid (22:6, the main component of fish oil), the vinyl ether-linked residues are usually otherwise saturated (i.e., there are no double bonds in the chain other than the vinyl ether group) or otherwise monounsaturated (i.e., there is one double bond in the chain in addition to the vinyl ether group).

[0004] Plasmalogen biosynthesis is a complex process involving multiple enzymes in peroxisomes and the endoplasmic reticulum. The rate-limiting step in this pathway is the formation of long-chain fatty alcohols by fatty acyl-CoA reductases 1 and 2 (Far-1 / 2). Oral administration of naturally occurring alkylglycerols (1-O-alkylglycerol or 1-O-alkyl-2,3-diacylglycerol) can bypass this rate-limiting step in plasmalogen synthesis. These are directly incorporated into the phospholipid pathway, bypassing the peroxisome. This increases circulating and tissue plasmalogen. Although alkylglycerols are present in our diet, typical dietary levels are insufficient to significantly increase plasmalogen levels in humans. Shark liver oil is rich in alkylglycerols and is currently used as a dietary supplement to reduce inflammation and improve immune function. Alkylglycerols can also be synthesized, providing a future avenue for environmentally sustainable sources of these compounds (Magnusson, CD, et al., Tetrahedron (2011) 67:1821-36; Shi, Y., et al., Green Chemistry (2010) 12(12)).

[0005] WO 2021 / 007623 A1 to Baker Heart and Diabetes Institute, which is incorporated herein by reference in its entirety, generally relates to various compositions and methods for maintaining or regulating the mixture of ether lipid molecules in human tissues for use in the treatment of non-disease states as well as certain disease states, including obesity, diabetes, fatty liver disease, cardiovascular disease, and Alzheimer's disease.

[0006] There is an unmet need for improved mixtures or compositions comprising precursor compounds for plasmalogens, which would serve as important contributions to one or more fields, including, but not limited to, medicinal, pharmaceutical, nutraceutical, and nutritional technology. Summary of the Invention

[0007] In one embodiment, a formulation is described that may take the form of one or more of a drug, pharmaceutical, dietary supplement, and food or other nutritional composition, and the formulation comprises, consists of, or consists essentially of one or more ether lipid compounds. It has been found that ingesting lysoalkylphosphatidylcholine (also known as LPC(O)) and alkylphosphatidylcholine (also known as PC(O)) in krill oil increases PE(P) and PC(P) species. This is not obvious, as it is generally believed that LPC(O) and PC(O) are dead ends in the plasmalogen biosynthetic pathway and that there is no active pathway for converting LPC(O) and PC(O) to PE(P) and PC(P). Therefore, it appears that LPC(O) / PC(O) can be shunted into the plasmalogen biosynthetic pathway and / or directly stimulate the natural biosynthetic pathway to increase plasmalogen synthesis.

[0008] Comparing the increase in circulating (plasma) plasmalogen levels after a given treatment with LPC(O) and PC(O) (krill oil) to the increase after a similar treatment with AKDAG (shark liver oil), it was determined that LPC(O) and PC(O) were more bioavailable. That is, it was surprisingly discovered that lower doses of LPC(O) and PC(O) could result in an increase in circulating plasmalogen levels similar to that obtained by administering relatively higher doses of AKDAG (shark liver oil).

[0009] It is noteworthy that the phospholipids found in krill oil are substantially different from those in humans, but there is hope that LPC(O) levels may be related to plasmalogens or at least involved in regulating plasmalogens in humans.

[0010] Furthermore, it was recognized that the alkyl chain composition in LPC(O) and PC(O) in krill oil differs substantially from the alkenyl chain composition in human plasmalogens, potentially affecting the composition of the resulting plasmalogen, as previously reported for alkylglycerols (AKG) and alkyldiacylglycerols (AKDAG) (WO 2021 / 007623). This has led to the present development and disclosure of formulations of LPC(O) and predicted formulations of lysoalkylphosphatidylethanolamine (also known as LPE(O)) or lysoalkylphosphatidic acid (also known as LPA(O)), as described herein, that match the known plasmalogen composition in human plasma.

[0011] Further aspects of the present disclosure will be more readily understood upon review of the following detailed description of various embodiments thereof when considered in conjunction with the accompanying drawings. [Brief explanation of the drawings]

[0012] [Figure 1] Figure 1 shows the change (percent change) in plasma ether lipid classes from baseline / pretreatment after supplementation with krill oil (KO), fish oil (FO), or shark liver oil (SLO). Changes in plasma ether lipid classes were grouped by supplementation. Subjects showed significant percent changes in ether lipid classes after KO (measured on days 15 and 30) and SLO treatment (measured on day 21). FO supplementation did not significantly affect plasma ether lipid classes. Nominal significance of treatment effects was determined using paired t-tests (krill oil baseline vs. krill oil day 15, krill oil baseline vs. krill oil day 30, fish oil baseline vs. fish oil day 15, fish oil baseline vs. fish oil day 30, SLO pretreatment vs. SLO day 21), where * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001. [Figure 2]Figure 1 shows the change (percent change) in plasma ether lipid classes from baseline / pretreatment after KO, FO, or SLO supplementation (grouped by lipid class). The effect of different supplements was grouped by plasma ether lipid class. Subjects showed significant percent changes in ether lipid classes after KO (measured on days 15 and 30) and SLO treatment (measured on day 21). FO supplementation did not significantly affect plasma ether lipid classes. The nominal significance of treatment effects was determined using paired t-tests (krill oil baseline vs. krill oil 15 days, krill oil baseline vs. krill oil 30 days, fish oil baseline vs. fish oil 15 days, fish oil baseline vs. fish oil 30 days, SLO pretreatment vs. SLO 21 days), where * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001. [Figure 3] The effect of KO supplementation on the alkenyl chain composition of PE plasmalogens is illustrated. Data are presented as mean ± SD. KO supplementation did not significantly affect the alkenyl chain composition of PE plasmalogens. The nominal significance of treatment effects was determined using repeated measures ANOVA followed by Tukey's multiple comparison test; * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001. [Figure 4] The effect of FO supplementation on the alkenyl chain composition of PE plasmalogens is illustrated. Data are presented as mean ± SD. FO supplementation did not significantly affect the alkenyl chain composition of PE plasmalogens. The nominal significance of treatment effects was determined using repeated measures ANOVA followed by Tukey's multiple comparison test; * indicates P < 0.05. [Figure 5] The effect of SLO supplementation on the alkenyl chain composition of PE plasmalogens is illustrated. Data are presented as mean ± SD. SLO supplementation significantly altered the alkenyl chain composition of PE plasmalogens. The proportion of 18:1 alkenyl chains containing PE plasmalogens increased, and the proportion of 16:0 and 18:0 alkenyl chains containing PE plasmalogens decreased. The nominal significance of treatment effects was determined using a paired t-test, with ** indicating P<0.01 and *** indicating P<0.001. [Figure 6] The effect of KO supplementation on the acyl chain composition of PE plasmalogens is illustrated. Data are presented as mean ± SD. KO supplementation decreased the proportion of 18:1, 18:2, and 20:4 containing PE plasmalogens and increased the proportion of 20:5 and 22:6 containing PE plasmalogens. The nominal significance of treatment effects was determined using repeated measures ANOVA followed by Tukey's multiple comparison test; ** indicates P<0.01, and *** indicates P<0.001. [Figure 7] The effect of FO supplementation on the acyl chain composition of PE plasmalogens is illustrated. Data are presented as mean ± SD. FO supplementation decreased the proportion of 18:1, 18:2, and 20:4 containing PE plasmalogens and increased the proportion of 20:5 and 22:6 containing PE plasmalogens. The nominal significance of treatment effects was determined using repeated measures ANOVA followed by Tukey's multiple comparison test; * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001. [Figure 8] The effect of SLO supplementation on the acyl chain composition of PE plasmalogens is illustrated. Data are presented as mean ± SD. SLO supplementation reduced 20:4 containing PE plasmalogens and increased the proportion of 18:1 and 22:6 containing PE plasmalogens. Nominal significance of treatment effects was determined using a paired t-test, with ** indicating P<0.01 and *** indicating P<0.001. [Figure 9] HepG2 cells before and after plasmalogen precursor treatment are shown. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursors for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). Images were taken at 4x magnification using an Olympus CKX41 inverted microscope and analyzed using ImageJ. Scale bars for all images are 400 μm. [Figure 10A]Figure 1 shows labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations in HepG2 cells after plasmalogen precursor treatment. A bar graph representation of labeled PE(P) concentrations for 16:0, 18:0, and 18:1 species is shown. Mean ± standard deviation (n = 3 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p > 0.05, ** indicates p ≤ 0.01, *** indicates p ≤ 0.001, and **** indicates p ≤ 0.0001. [Figure 10B] The graph shows the concentration of labeled phosphatidylethanolamine plasmalogen (PE(P)) after plasmalogen precursor treatment in HepG2 cells. The ratio of labeled PE(P) to total labeling (%) is shown. Total labeling refers to the sum of labeled AKG, alkyl-acylglycerol (DG(O)), lysoalkylphosphatidylcholine (LPC(O)), lysophosphatidylcholine plasmalogen (LPC(P)), lysoalkylphosphatidylethanolamine (LPE(O)), lysophosphatidylethanolamine plasmalogen (LPE(P)), alkylphosphatidylcholine (PC(O)), phosphatidylcholine plasmalogen (PC(P)), alkylphosphatidylethanolamine (PE(O)), PE(P), and monoalkyl-diacylglycerol (TG(O)). The mean ± standard deviation (n = 3 per group) is shown, and each symbol represents an individual sample. Controls in which background values ​​are divided by background values ​​were excluded because they can be misleading. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 11]Figure 1 shows label incorporation into lipid classes of the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in HepG2 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). Concentrations were normalized to phosphatidylcholine (PC) 34:1, one of the most abundant endogenous PCs. Mean ± standard deviation (n = 3 per group) is shown, and each symbol represents an individual sample. (A) Bar graph showing label concentration after AKG treatment. (B) Bar graph showing label concentration after LPC(O) treatment. (C) Bar graph showing label concentration after LPE(O) treatment. [Figure 12A] Distribution of label incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in HepG2 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). Label concentration of each lipid class divided by total label is shown as a percentage of AKG treatment. Total labeling refers to the sum of labeled AKG, alkyl-acylglycerol (DG(O)), lysoalkylphosphatidylcholine (LPC(O)), lysophosphatidylcholine plasmalogen (LPC(P)), lysoalkylphosphatidylethanolamine (LPE(O)), lysophosphatidylethanolamine plasmalogen (LPE(P)), alkylphosphatidylcholine (PC(O)), phosphatidylcholine plasmalogen (PC(P)), alkylphosphatidylethanolamine (PE(O)), phosphatidylethanolamine plasmalogen (PE(P)), and monoalkyl-diacylglycerol (TG(O)). [Figure 12B]Distribution of label incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in HepG2 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). The label concentration of each lipid class divided by the total label is shown as a percentage of LPC(O) treatment. Total labeling refers to the sum of labeled AKG, alkyl-acylglycerol (DG(O)), lysoalkylphosphatidylcholine (LPC(O)), lysophosphatidylcholine plasmalogen (LPC(P)), lysoalkylphosphatidylethanolamine (LPE(O)), lysophosphatidylethanolamine plasmalogen (LPE(P)), alkylphosphatidylcholine (PC(O)), phosphatidylcholine plasmalogen (PC(P)), alkylphosphatidylethanolamine (PE(O)), phosphatidylethanolamine plasmalogen (PE(P)), and monoalkyl-diacylglycerol (TG(O)). [Figure 12C]Distribution of label incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in HepG2 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). The label concentration of each lipid class divided by the total label is shown as a percentage of LPE(O) treatment. Total labeling refers to the sum of labeled AKG, alkyl-acylglycerol (DG(O)), lysoalkylphosphatidylcholine (LPC(O)), lysophosphatidylcholine plasmalogen (LPC(P)), lysoalkylphosphatidylethanolamine (LPE(O)), lysophosphatidylethanolamine plasmalogen (LPE(P)), alkylphosphatidylcholine (PC(O)), phosphatidylcholine plasmalogen (PC(P)), alkylphosphatidylethanolamine (PE(O)), phosphatidylethanolamine plasmalogen (PE(P)), and monoalkyl-diacylglycerol (TG(O)). [Figure 13] 3T3 cells before and after plasmalogen precursor treatment are shown. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). Images were taken at 4x magnification using an Olympus CKX41 inverted microscope and analyzed using ImageJ. Scale bars for all images are 400 μm. [Figure 14A]Labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations after plasmalogen precursor treatment in 3T3 cells are shown. Concentrations were normalized to phosphatidylcholine (PC) 34:1, one of the most abundant endogenous PCs. A bar graph representation of labeled PE(P) concentrations for 16:0, 18:0, and 18:1 species is shown. Mean ± standard deviation (n = 3 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p > 0.05, ** indicates p ≤ 0.01, *** indicates p ≤ 0.001, and **** indicates p ≤ 0.0001. [Figure 14B] Figure 1 shows labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations after plasmalogen precursor treatment in 3T3 cells. Concentrations were normalized to phosphatidylcholine (PC) 34:1, one of the most abundant endogenous PCs. A bar graph displaying the log(2) of labeled PE(P) concentrations for 16:0, 18:0, and 18:1 species is shown. Mean ± standard deviation (n = 3 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p > 0.05, ** indicates p ≤ 0.01, *** indicates p ≤ 0.001, and **** indicates p ≤ 0.0001. [Figure 14C]Labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations after plasmalogen precursor treatment in 3T3 cells are shown. Concentrations were normalized to phosphatidylcholine (PC) 34:1, one of the most abundant endogenous PCs. The ratio of labeled PE(P) to total labeling (%) is shown. Total labeling refers to the sum of labeled AKG, alkyl-acylglycerol (DG(O)), lysoalkylphosphatidylcholine (LPC(O)), lysophosphatidylcholine plasmalogen (LPC(P)), lysoalkylphosphatidylethanolamine (LPE(O)), lysophosphatidylethanolamine plasmalogen (LPE(P)), alkylphosphatidylcholine (PC(O)), phosphatidylcholine plasmalogen (PC(P)), alkylphosphatidylethanolamine (PE(O)), PE(P), and monoalkyl-diacylglycerol (TG(O)). The mean ± standard deviation (n = 3 per group) is shown, and each symbol represents an individual sample. Controls in which background values ​​are divided by background values ​​were excluded because they could be misleading. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p > 0.05, ** indicates p ≤ 0.01, *** indicates p ≤ 0.001, and **** indicates p ≤ 0.0001. [Figure 15] Figure 1 shows label incorporation into lipid classes of the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in 3T3 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). Concentrations were normalized to phosphatidylcholine (PC) 34:1, one of the most abundant endogenous PCs. Mean ± standard deviation (n = 3 per group) is shown, and each symbol represents an individual sample. (A) Bar graph showing label concentration after AKG treatment. (B) Bar graph showing label concentration after LPC(O) treatment. (C) Bar graph showing label concentration after LPE(O) treatment. [Figure 16A]Figure 1 shows the distribution of label incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in 3T3 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). The label concentration of each lipid class divided by the total label is shown as a percentage of AKG treatment. [Figure 16B] Figure 1 shows the distribution of label incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in 3T3 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). The label concentration of each lipid class divided by the total label is shown as a percentage of LPC(O) treatment. [Figure 16C] Figure 1 shows the distribution of label incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor treatment in 3T3 cells. Cells were treated with 0.05% ethanol (vehicle control) or 20 μM labeled plasmalogen precursor for 24 hours. Labeled precursors included alkylglycerol (O-16:0) d2 (AKG), LPC (O-16:0) d4 (LPC(O)), and LPE (O-16:0) d5 (LPE(O)). The label concentration of each lipid class divided by the total label is shown as a percentage of LPE(O) treatment. [Figure 17] We describe a procedure for administering a mixture of deuterated precursor molecules to sixty-four 8-week-old C57BL / 6 mice. [Figure 18A]Labeled alkylglycerol (AKG) concentrations after dose mixture A of plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A graph showing the concentrations of labeled AKG O-16:0 in female mice after dose mixture A of treatment is shown at time points 0, 1, 4, 24, and 48 hours. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 18B] Labeled alkylglycerol (AKG) concentrations after dose mixture A of plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the concentrations of labeled AKG O-16:0 in males after dose mixture A of treatment is shown at time points 0, 1, 4, 24, and 48 hours. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 19A]Labeled alkylphosphatidylcholine (PC(O)) concentrations after dose mixture A of plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A graph showing the concentrations of labeled PC(O) 16:0 species at time points 0, 1, 4, 24, and 48 hours after dose mixture A of treatment in females is shown. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 19B] Labeled alkylphosphatidylcholine (PC(O)) concentrations after dose mixture A of plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A graph showing the concentrations of labeled PC(O) 16:0 species at time points 0, 1, 4, 24, and 48 hours after dose mixture A of treatment in males is shown. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 20A]Figure 1 shows labeled alkylphosphatidylethanolamine (PE(O)) concentrations after Dose Mixture A of plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A graph is shown displaying the concentrations of labeled PE(O) 16:0 species at time points 0, 1, 4, 24, and 48 hours after Dose Mixture A of treatment in females. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 20B] Figure 1 shows labeled alkylphosphatidylethanolamine (PE(O)) concentrations after dose mixture A of plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A graph is shown displaying the concentrations of labeled PE(O) 16:0 species at time points 0, 1, 4, 24, and 48 hours after dose mixture A of treatment in males. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 21A]Figure 1 shows labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations after Dose Mixture A of plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A graph is shown displaying the concentrations of labeled PE(P) 16:0 species at time points 0, 1, 4, 24, and 48 hours after Dose Mixture A of treatment in females. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 21B] Figure 1 shows labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations after Dose Mixture A of plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A graph is shown displaying the concentrations of labeled PE(P) 16:0 species at time points 0, 1, 4, 24, and 48 hours after Dose Mixture A of treatment in males. The mean ± standard deviation (n = 4-5 per group) is shown, and each symbol represents an individual sample. [Figure 22A]Figure 1 shows labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations after dose mixture B of plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A graph is shown displaying the concentrations of labeled PE(P) 16:0 species at time points 0, 1, 4, 24, and 48 hours after dose mixture B of treatment in females. The mean ± standard deviation (n=4 per group) is shown, and each symbol represents an individual sample. [Figure 22B] Figure 1 shows labeled phosphatidylethanolamine plasmalogen (PE(P)) concentrations after dose mixture B of plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC(O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE(O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A graph showing the concentrations of labeled PE(P) 16:0 species at time points 0, 1, 4, 24, and 48 hours after dose mixture B treatment in males is shown. The mean ± standard deviation (n=4 per group) is shown, and each symbol represents an individual sample. [Figure 23A]Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the Cmax of treatment dose mixture A in females. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 23B] Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the log(2) Cmax of treatment dose mixture A in females. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 23C]Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the dose of treatment mixture A in males. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 23D] Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the log(2) Cmax of treatment dose mixture A in males. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 23E]Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the dose of treatment mixture B in female mice. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 23F] Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the log(2) Cmax of treatment with Mixture B in female mice. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 23G]Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the dose of treatment mixture B in males. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 23H] Figure 1 shows the maximum concentration (Cmax) of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the log(2) Cmax of treatment dose mixture B in males. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24A]Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displaying the AUC of treatment dose mixture A in female mice is shown. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24B] Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the log(2) AUC of treatment dose mixture A in females is shown. The mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24C]Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displaying the AUC of treatment dose mixture A in males is shown. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24D] Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the log(2) AUC of treatment dose mixture A in males is shown. The mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24E]Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displays the AUC of treatment dose mixture B in female mice. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24F] Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the log(2) AUC of treatment dose mixture B in females is shown. The mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24G]Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours later. A bar graph displaying the AUC of treatment dose mixture B in males is shown. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 24H] Area under the curve (AUC) values ​​of labeled phosphatidylethanolamine plasmalogen after plasmalogen precursor treatment in mice are shown. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the log(2) AUC of treatment dose mixture B in males is shown. The mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA with Tukey's post-hoc test was used. ns indicates p>0.05, * indicates p≦0.05, ** indicates p≦0.01, *** indicates p≦0.001, and **** indicates p≦0.0001. [Figure 25A]Figure 1 shows the area under the curve (AUC) of lipid classes in the plasmalogen biosynthetic pathway after dose mixture A of plasmalogen precursor treatment in female mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), or LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the AUC of AKG treatment is shown. The mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. [Figure 25B] Figure 1 shows the area under the curve (AUC) of lipid classes in the plasmalogen biosynthetic pathway after dose mixture A of plasmalogen precursor treatment in female mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), or LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the AUC of LPC(O) treatment is shown. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. [Figure 25C]Figure 1 shows the area under the curve (AUC) of lipid classes in the plasmalogen biosynthetic pathway after dose mixture A of plasmalogen precursor treatment in female mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), or LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleeding before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying the AUC of LPE(O) treatment is shown. The mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. [Figure 26A] Figure 1 shows the area under the curve (AUC) of lipid classes in the plasmalogen biosynthetic pathway after dose mixture A of plasmalogen precursor treatment in male mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), or LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying AKG treatment is shown. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. [Figure 26B]Figure 1 shows the area under the curve (AUC) of lipid classes in the plasmalogen biosynthetic pathway after dose mixture A of plasmalogen precursor treatment in male mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), or LPE(O) mixture (LPE (O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying LPC(O) treatment is shown. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. [Figure 26C] Figure 1 shows the area under the curve (AUC) of lipid classes in the plasmalogen biosynthetic pathway after dose mixture A of plasmalogen precursor treatment in male mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG (O-16:0, 18:0, 18:1) d2), LPC(O) mixture (LPC (O-16:0, 18:0, 18:1) d2), or LPE(O) mixture (LPE(O-16:0) d5 and unlabeled LPE (O-16:0, 18:0, 18:1)). Blood was collected by tail bleed before treatment and 1, 4, 24, and 48 hours after treatment. A bar graph displaying LPE(O) treatment is shown. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. [Figure 27]Figure 1 shows the O-16:0 species of labeled phosphatidylethanolamine plasmalogen in the brain after supplementation with labeled LPE(O) in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control) or an LPE(O) mixture (LPE(O-16:0, O-18:0, O-18:1, O-16:0-d5)). Brain samples were collected 48 hours after gavage. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. Student's t-test was used to compare mean differences between groups. * indicates p ≤ 0.05, and *** indicates p ≤ 0.001. (A) Bar graph showing treatment dose mixture A in females. (B) Bar graph showing treatment dose mixture A in males. (C) Bar graph showing treatment dose mixture B in females. (D) Bar graph showing treatment dose mixture B in males. [Figure 28] Figure 1 shows the O-16:0 species of labeled phosphatidylethanolamine plasmalogen in the brain after supplementation with labeled AKG or LPC(O) in mice. Eight-week-old C57BL / 6 mice were gavaged with a single dose of lecithin (vehicle control), AKG mixture (AKG(O-16:0-d2, O-18:0-d2, O-18:1-d2)), or LPC(O) mixture (LPC(O-16:0-d2, O-18:0-d2)). Brain samples were collected 48 hours after gavage. Mean ± standard deviation (n = 4 per group) is shown, and each symbol represents an individual sample. One-way ANOVA was used to test mean differences between groups. (A) Bar graph showing treatment dose mixture A in females. (B) Bar graph showing treatment dose mixture A in males. (C) Bar graph showing treatment dose mixture B in females. (D) Bar graph displaying dose mixture B of treatment in males. [Figure 29A] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of precursor compound AKG-dose mixture C. [Figure 29B] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture C. [Figure 29C] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of the precursor compound AKDAG-DHA-dose mixture C. [Figure 29D] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of precursor compound LPC(O)-dose mixture C. [Figure 29E] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of precursor compound AKG-dose mixture D. [Figure 29F] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture E. [Figure 29G] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of the precursor compound AKDAG-DHA-dose mixture D. [Figure 29H] 1 shows the change in tracer alkyldiacylglycerol [TG(O)] concentration after administration of precursor compound LPC(O)-dose mixture D. [Figure 30A] 1 shows the change in the concentration of the tracer lysoalkylphosphatidylcholine [LPC(O)] after administration of the precursor compound AKG-dose mixture C. [Figure 30B] 1 shows the change in the concentration of the tracer lysoalkylphosphatidylcholine [LPC(O)] after administration of the precursor compound AKDAG-oleic acid-dose mixture C. [Figure 30C] 1 shows the change in the concentration of the tracer lysoalkylphosphatidylcholine [LPC(O)] after administration of the precursor compound AKDAG-DHA-dose mixture C. [Figure 30D] 1 shows the change in tracer lysoalkylphosphatidylcholine [LPC(O)] concentration after administration of precursor compound LPC(O)-dose mixture C. [Figure 30E] 1 shows the change in the concentration of the tracer lysoalkylphosphatidylcholine [LPC(O)] after administration of the precursor compound AKG-dose mixture D. [Figure 30F] 1 shows the change in the concentration of the tracer lysoalkylphosphatidylcholine [LPC(O)] after administration of the precursor compound AKDAG-oleic acid-dose mixture E. [Figure 30G] 1 shows the change in the concentration of the tracer lysoalkylphosphatidylcholine [LPC(O)] after administration of the precursor compound AKDAG-DHA-dose mixture D. [Figure 30H] 1 shows the change in tracer lysoalkylphosphatidylcholine [LPC(O)] concentration after administration of precursor compound LPC(O)-dose mixture D. [Figure 31A] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of precursor compound AKG-dose mixture C. [Figure 31B] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture C. [Figure 31C] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of precursor compound AKDAG-DHA-dose mixture C. [Figure 31D] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of precursor compound LPC(O)-dose mixture C. [Figure 31E] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of precursor compound AKG-dose mixture D. [Figure 31F] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture E. [Figure 31G] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of precursor compound AKDAG-DHA-dose mixture D. [Figure 31H] 1 shows the change in tracer alkylphosphatidylcholine [PC(O)] concentration after administration of precursor compound LPC(O)-dose mixture D. [Figure 32A] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of precursor compound AKG-dose mixture C. [Figure 32B] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture C. [Figure 32C] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of the precursor compound AKDAG-DHA-dose mixture C. [Figure 32D] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of precursor compound LPC(O)-dose mixture C. [Figure 32E] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of precursor compound AKG-dose mixture D. [Figure 32F] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture E. [Figure 32G] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of precursor compound AKDAG-DHA-dose mixture D. [Figure 32H] 1 shows the change in tracer alkylphosphatidylethanolamine [PE(O)] concentration after administration of precursor compound LPC(O)-dose mixture D. [Figure 33A] 1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of the precursor compound AKG-dose mixture C. [Figure 33B] FIG. 1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture C. [Figure 33C]1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of the precursor compound AKDAG-DHA-dose mixture C. [Figure 33D] 1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of precursor compound LPC(O)-dose mixture C. [Figure 33E] 1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of precursor compound AKG-dose mixture D. [Figure 33F] FIG. 1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture E. [Figure 33G] 1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of the precursor compound AKDAG-DHA-dose mixture D. [Figure 33H] 1 shows the change in tracer alkenylphosphoethanolamine [PE(P)] concentration after administration of precursor compound LPC(O)-dose mixture D. [Figure 34A] 1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of precursor compound AKG-dose mixture C. [Figure 34B] FIG. 1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture C. [Figure 34C] 1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of the precursor compound AKDAG-DHA-dose mixture C. [Figure 34D] 1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of precursor compound LPC(O)-dose mixture C. [Figure 34E] 1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of precursor compound AKG-dose mixture D. [Figure 34F]1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of the precursor compound AKDAG-oleic acid-dose mixture E. [Figure 34G] 1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of precursor compound AKDAG-DHA-dose mixture D. [Figure 34H] 1 shows the change in tracer alkenylphosphatidylcholine [PC(P)] concentration after administration of precursor compound LPC(O)-dose mixture D. [Figure 35A] Figure 1 shows the maximum plasma concentrations (Cmax) of the major lipid classes after administration of the precursor compound TG(O). [Figure 35B] Figure 1 shows the maximum plasma concentrations (Cmax) of the major lipid classes after administration of the precursor compound LPC(O). [Figure 35C] Figure 1 shows the maximum plasma concentrations (Cmax) of the major lipid classes after administration of the precursor compound PC(O). [Figure 35D] Figure 1 shows the maximum plasma concentrations (Cmax) of the major lipid classes after administration of the precursor compound PE(O). [Figure 35E] Figure 1 shows the maximum plasma concentrations (Cmax) of the major lipid classes after administration of the precursor compound PE (P). [Figure 35F] Figure 1 shows the maximum plasma concentrations (Cmax) of the major lipid classes after administration of the precursor compound PC (P). [Figure 36A] Figure 1 shows the area under the curve (AUC) of the major lipid classes after administration of the precursor compound TG(O). [Figure 36B] Figure 1 shows the area under the curve (AUC) of the major lipid classes after administration of the precursor compound LPC(O). [Figure 36C] Figure 1 shows the area under the curve (AUC) of the major lipid classes after administration of the precursor compound PC(O). [Figure 36D] Figure 1 shows the area under the curve (AUC) of the major lipid classes after administration of the precursor compound PE(O). [Figure 36E] Figure 1 shows the area under the curve (AUC) of the major lipid classes after administration of the precursor compound PE (P). [Figure 36F] Figure 1 shows the area under the curve (AUC) of the major lipid classes after administration of the precursor compound PC (P). [Figure 37A] 1 shows the effect of labeled precursor TG(O) supplementation on major ether lipid classes in the liver. [Figure 37B] 1 shows the effect of labeled precursor LPC(O) supplementation on major ether lipid classes in liver. [Figure 37C] 1 shows the effect of labeled precursor PC(O) supplementation on major ether lipid classes in liver. [Figure 37D] 1 shows the effect of labeled precursor PE(O) supplementation on major ether lipid classes in liver. [Figure 37E] 1 shows the effect of labeled precursor PE(P) supplementation on major ether lipid classes in liver. [Figure 37F] 1 shows the effect of labeled precursor PC(P) supplementation on major ether lipid classes in liver. [Figure 38A] 1 shows the effect of labeled precursor TG(O) supplementation on major ether lipid classes in the spleen. [Figure 38B] 1 shows the effect of labeled precursor LPC(O) supplementation on major ether lipid classes in the spleen. [Figure 38C] 1 shows the effect of labeled precursor PC(O) supplementation on major ether lipid classes in the spleen. [Figure 38D] 1 shows the effect of labeled precursor PE(O) supplementation on major ether lipid classes in the spleen. [Figure 38E] 1 shows the effect of labeled precursor PE(P) supplementation on major ether lipid classes in the spleen. [Figure 38F] 1 shows the effect of labeled precursor PC(P) supplementation on major ether lipid classes in the spleen. [Figure 39A] 1 shows the effect of labeled precursor TG(O) supplementation on major ether lipid classes in the brain. [Figure 39B] 1 shows the effect of labeled precursor LPC(O) supplementation on major ether lipid classes in the brain. [Figure 39C] 1 shows the effect of labeled precursor PC(O) supplementation on major ether lipid classes in the brain. [Figure 39D] 1 shows the effect of supplementation with the labeled precursor PE(O) on major ether lipid classes in the brain. [Figure 39E] 1 shows the effect of supplementation with the labeled precursor PE(P) on major ether lipid classes in the brain. [Figure 39F] 1 shows the effect of labeled precursor PC(P) supplementation on major ether lipid classes in the brain. [Figure 40A] 1 shows the effect of labeled precursor TG(O) supplementation on major ether lipid classes in the kidney. [Figure 40B] 1 shows the effect of labeled precursor LPC(O) supplementation on major ether lipid classes in the kidney. [Figure 40C] 1 shows the effect of labeled precursor PC(O) supplementation on major ether lipid classes in the kidney. [Figure 40D] 1 shows the effect of labeled precursor PE(O) supplementation on major ether lipid classes in the kidney. [Figure 40E] 1 shows the effect of labeled precursor PE(P) supplementation on major ether lipid classes in the kidney. [Figure 40F] 1 shows the effect of labeled precursor PC(P) supplementation on major ether lipid classes in the kidney. [Figure 41A] 1 shows the effect of labeled precursor TG(O) supplementation on major ether lipid classes in visceral adipose tissue. [Figure 41B] 1 shows the effect of labeled precursor LPC(O) supplementation on major ether lipid classes in visceral adipose tissue. [Figure 41C] 1 shows the effect of labeled precursor PC(O) supplementation on major ether lipid classes in visceral adipose tissue. [Figure 41D] 1 shows the effect of supplementation with the labeled precursor PE(O) on major ether lipid classes in visceral adipose tissue. [Figure 41E]Figure 1 shows the effect of labeled precursor PE(P) supplementation on major ether lipid classes in visceral adipose tissue. [Figure 41F] 1 shows the effect of labeled precursor PC(P) supplementation on major ether lipid classes in visceral adipose tissue. [Figure 42A] 1 shows the effect of labeled precursor TG(O) supplementation on major ether lipid classes in skeletal muscle. [Figure 42B] 1 shows the effect of supplementation with the labeled precursor LPC(O) on major ether lipid classes in skeletal muscle. [Figure 42C] 1 shows the effect of labeled precursor PC(O) supplementation on major ether lipid classes in skeletal muscle. [Figure 42D] 1 shows the effect of supplementation with the labeled precursor PE(O) on major ether lipid classes in skeletal muscle. [Figure 42E] 1 shows the effect of supplementation with the labeled precursor PE(P) on major ether lipid classes in skeletal muscle. [Figure 42F] 1 shows the effect of labeled precursor PC(P) supplementation on major ether lipid classes in skeletal muscle. [Figure 43A] 1 shows the effect of labeled precursor TG(O) supplementation on major ether lipid classes in the heart. [Figure 43B] 1 shows the effect of supplementation with the labeled precursor LPC(O) on major ether lipid classes in the heart. [Figure 43C] 1 shows the effect of labeled precursor PC(O) supplementation on major ether lipid classes in the heart. [Figure 43D] 1 shows the effect of supplementation with the labeled precursor PE(O) on major ether lipid classes in the heart. [Figure 43E] 1 shows the effect of supplementation with the labeled precursor PE(P) on major ether lipid classes in the heart. [Figure 43F] 1 shows the effect of labeled precursor PC(P) supplementation on major ether lipid classes in the heart. [Figure 44]Figure 1 shows the effect of LPC(O) supplementation on plasmalogen levels in RAW264.7 cells. RAW264.7 macrophage cells were treated with 20 μM LPC(O) with different SN1 compositions for 24 hours and then harvested for lipidomic analysis. PE-plasmalogen or PE(P) data were normalized to total cellular phosphatidylcholine (PC) levels and presented as mean ± SD (n = 3 / group). Each circle represents an individual data point. Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. * indicates P < 0.05, ** indicates P < 0.01, and **** indicates P < 0.0001 compared to control. [Figure 45] The effect of LPE(O) supplementation on plasmalogen levels in RAW264.7 cells is shown. RAW264.7 macrophage cells were treated with 20 μM LPE(O) with different SN1 compositions for 24 hours and then harvested for lipidomic analysis. PE-plasmalogen or PE(P) data were normalized to total cellular phosphatidylcholine (PC) levels and presented as mean ± SD (n = 3 / group). Each circle represents an individual data point. Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. **** indicates P < 0.0001 compared to control. [Figure 46] The effect of LPC(O) supplementation on plasmalogen composition in RAW264.7 cells is shown. RAW264.7 macrophage cells were treated with 20 μM LPC(O) containing different SN1 compositions for 24 h and then harvested for lipidomic analysis. The relative proportions of different SN1 containing PE(P) are presented as mean ± SD (n = 3 / group). [Figure 47]The effect of LPC(O) supplementation on plasmalogen levels in 3T3-L1 cells is shown. 3T3-L1 preadipocytes were treated with 20 μM LPC(O) with different SN1 compositions for 24 hours and then harvested for lipidomic analysis. PE-plasmalogen or PE(P) data were normalized to total cellular phosphatidylcholine (PC) levels and presented as mean ± SD (n = 3 / group). Each circle represents an individual data point. Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. * indicates P < 0.05, ** indicates P < 0.01, and **** indicates P < 0.0001 compared to control. [Figure 48] The effect of LPE(O) supplementation on plasmalogen levels in 3T3-L1 cells is shown. 3T3-L1 preadipocytes were treated with 20 μM LPE(O) with different SN1 compositions for 24 hours and then harvested for lipidomic analysis. PE-plasmalogen or PE(P) data were normalized to total cellular phosphatidylcholine (PC) levels and presented as mean ± SD (n = 3 / group). Each circle represents an individual data point. Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001, and **** indicates P < 0.0001 compared to control. [Figure 49] The effect of LPC(O) supplementation on plasmalogen composition in 3T3-L1 cells is shown. 3T3-L1 cells were treated with 20 μM LPC(O) containing different SN1 compositions for 24 hours and then harvested for lipidomic analysis. The relative proportions of different SN1 containing PE(P) are presented as mean ± SD (n = 3 / group). [Figure 50] The effect of LPE(O) supplementation on plasmalogen composition in 3T3-L1 cells is shown. 3T3-L1 cells were treated with 20 μM LPE(O) containing different SN1 compositions for 24 h and then harvested for lipidomic analysis. The relative proportions of different SN1 containing PE(P) are presented as mean ± SD (n = 3 / group). [Figure 51A]Figure 1 shows label incorporation (area under the curve) into lipid classes of the plasmalogen biosynthetic pathway after low dose plasmalogen precursor supplementation. Figure 2 shows incorporation after AKG supplementation. [Figure 51B] Figure 1 shows label incorporation (area under the curve) into lipid classes of the plasmalogen biosynthetic pathway after low doses of plasmalogen precursor supplementation. Figure 2 shows incorporation after AKDAG-oleic acid supplementation. [Figure 51C] Figure 1 shows label incorporation (area under the curve) into lipid classes of the plasmalogen biosynthetic pathway after low dose plasmalogen precursor supplementation. Figure 2 shows incorporation after AKDAG-DHA supplementation. [Figure 51D] Figure 1 shows label incorporation (area under the curve) into lipid classes of the plasmalogen biosynthetic pathway after low dose plasmalogen precursor supplementation. Figure 2 shows incorporation after LPC(O) supplementation. [Figure 52A] 1 shows the distribution % of labeled incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor supplementation. Incorporation after AKG supplementation is shown. [Figure 52B] 1 shows the % distribution of labeled incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor supplementation. Incorporation after AKDAG-oleic acid supplementation is shown. [Figure 52C] 1 shows the distribution % of labeled incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor supplementation. 1 shows the incorporation after AKDAG-DHA supplementation. [Figure 52D] Figure 1 shows the distribution % of labeled incorporation in the plasmalogen biosynthetic pathway after plasmalogen precursor supplementation. Figure 2 shows incorporation after LPC(O) supplementation. [Figure 53A] Labeled ether lipids in feces after 24 hours of labeled precursor TG(O) supplementation are shown. [Figure 53B] Labeled ether lipids in feces after 24 hours of supplementation with the labeled precursor LPC(O) are shown. [Figure 53C] Labeled ether lipids in feces after 24 hours of labeled precursor PC(O) supplementation are shown. [Figure 53D] Labeled ether lipids in feces after 24 hours of supplementation with the labeled precursor PE(O) are shown. [Figure 53E] Labeled ether lipids in feces after 24 hours of supplementation with the labeled precursor PE(P) are shown. [Figure 53F] Labeled ether lipids in feces after 24 hours of labeled precursor PC(P) supplementation are shown. [Figure 54A] Labeled ether lipids in feces after 24 hours of labeled precursor TG(O) supplementation are shown. [Figure 54B] Labeled ether lipids in feces after 24 hours of supplementation with the labeled precursor LPC(O) are shown. [Figure 54C] Labeled ether lipids in feces after 24 hours of labeled precursor PC(O) supplementation are shown. [Figure 54D] Labeled ether lipids in feces after 24 hours of supplementation with the labeled precursor PE(O) are shown. [Figure 54E] Labeled ether lipids in feces after 24 hours of supplementation with the labeled precursor PE(P) are shown. [Figure 54F] Labeled ether lipids in feces after 24 hours of labeled precursor PC(P) supplementation are shown. [Figure 55] Figure 1 shows that all LPX(O) treatments suppressed the LPS-induced increase in TLR4 gene expression. Data are normalized to control + LPS. Data are presented as mean ± SD (n = 3 / group). Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. An asterisk (*) indicates P < 0.05 compared to the control group, and a ^ indicates P < 0.05 compared to the control group + LPS. [Figure 56] Figure 1 shows that all LPX(O) treatments suppressed the LPS-induced increase in TLR4 gene expression. Data are normalized to control. Data are presented as mean ± SD (n = 3 / group). Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. * indicates P < 0.05 compared to control, and ^ indicates P < 0.05 compared to control + LPS. [Table 1] DETAILED DESCRIPTION OF THE INVENTION

[0013] The general structure of the phospholipids described herein is represented by the following formula (I): [ka]

[0014] In compounds of formula (I), the sn-1 carbon is labeled as above. 3 R includes phosphates and substituted phosphates. 1 and R 2 can each independently be hydrogen, alkyl, alkenyl, acyl, etc., or a substituted version of these moieties as described herein.

[0015] In Scheme I, some exemplary plasmalogens are shown in the top two structures, with boxes highlighting the sn-1 substitutions from formula (I). [ka] Scheme I. Typical plasmalogens.

[0016] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0017] The term "and / or," e.g., "X and / or Y," shall be understood to mean "X and Y" or "X or Y," and shall be deemed to provide explicit support for both meanings or either meaning.

[0018] As used herein, the words "a" and "an" mean "one or more" and include the plural forms unless the context is inappropriate.

[0019] As used herein, the term "alkyl" refers to a straight-chain or branched group of 1 to 30, 1 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atoms (referred to herein as C1 to C6, respectively). 30 Alkyl, C1-C 18 Alkyl, C 14 ~C 24 Alkyl, C 14 ~C 18 Alkyl, C 16 Alkyl or C 18 "Alkyl" refers to saturated straight chain or branched hydrocarbon groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, isobutyl, t-butyl, isopentyl, neopentyl, and the like.

[0020] As used herein, the term "alkenyl" refers to a straight-chain or branched group of 2 to 30, 2 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atoms (respectively defined herein as C2 to C6). 30 Alkenyl, C2-C 18 Alkenyl, C 14 ~C 24 Alkenyl, C 14 ~C 18 Alkenyl, C 16 Alkenyl, or C 18Alkenyl refers to unsaturated straight-chain or branched hydrocarbons such as olefins (also referred to as alkenyl). Alkenyl compounds may have multiple double bonds. Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, isopropenyl, 2-methyl-1-butenyl, 3-methyl-1-butenyl, 2-methyl-3-butenyl, 2,2-dimethyl-1-propenyl, and 2,2-dimethyl-1-propenyl. Examples of alkenyl include, but are not limited to, 2-methyl-1-pentenyl, 3-methyl-1-pentenyl, 4-methyl-1-pentenyl, 2-methyl-2-pentenyl, 3-methyl-2-pentenyl, 4-methyl-2-pentenyl, 2,2-dimethyl-1-butenyl, 3,3-dimethyl-1-butenyl, 2-ethyl-1-butenyl, isobutenyl, t-butenyl, isopentenyl, neopentenyl, and the like. The term "alkenyl" can also be interpreted as meaning an alkene of structure C18:1 in which the double bond is typically at the n7 or n9 position. The term "alkenyl" can also be interpreted as meaning an alkene of structure C18:1 in which the double bond is between the first and second carbons adjacent to the ether bond. The term "alkenyl" can also be interpreted as meaning an alkene in which one double bond is at the n7 or n9 position and a second double bond is between the first and second carbons.

[0021] The term "acyl," as used herein, refers to a radical of the general formula -C(O)R, where R is hydrogen or a straight or branched chain group of 1 to 30, 1 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atoms (referred to herein as C1 to C5, respectively). 30 Alkyl, C1-C 18 Alkyl, C 14 ~C 24 Alkyl, C 14 ~C 18 Alkyl, C 16 Alkyl or C 18alkyl), or straight or branched groups of 2 to 30, 2 to 18, 14 to 24, 14 to 18, 16 or 18 carbon atoms (referred to herein as C2 to C6, respectively). 30 Alkenyl, C2-C 18 Alkenyl, C 14 ~C 24 Alkenyl, C 14 ~C 18 Alkenyl, C 16 Alkenyl, or C 18 The alkyl groups are saturated or unsaturated, straight chain or branched hydrocarbons, such as alkyl groups (also called alkenyl groups). Exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, isobutyl, t-butyl, isopentyl, neopentyl, and the like.

[0022] The term "ether," as used herein, refers to any organic compound having a structure similar to an ether functional group and having an oxygen atom connecting two alkyl or other organic groups. As used herein, the term "ether" refers to a straight or branched chain group of 1 to 30, 1 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atoms (referred to herein as C1 to C6, respectively). 30 Alkyl, C1-C 18 Alkyl, C 14 ~C 24 Alkyl, C 14 ~C 18 Alkyl, C 16 Alkyl or C 18As used herein, the term "ether" may refer to a functional group having an oxygen atom linking a saturated straight chain or branched hydrocarbon, such as an alkyl, to a second saturated straight chain or branched hydrocarbon. As used herein, the term "ether" refers to a straight chain or branched group of 1 to 30, 1 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atoms (referred to herein as C1 to C6, respectively). 30 Alkyl, C1-C 18 Alkyl, C 14 ~C 24 Alkyl, C 14 ~C 18 Alkyl, C 16 Alkyl or C 18 saturated straight chain or branched hydrocarbons such as alkyl, and straight chain or branched groups of 2 to 30, 2 to 18, 14 to 24, 14 to 18, 16 or 18 carbon atoms (referred to herein as C2 to C6, respectively). 30 Alkenyl, C2-C 18 Alkenyl, C 14 ~C 24 Alkenyl, C 14 ~C 18 Alkenyl, C 16 Alkenyl, or C 18 As used herein, the term "ether" may refer to a functional group having an oxygen atom linking an unsaturated straight-chain or branched hydrocarbon, such as an alkyl group, to a straight-chain or branched group of 2 to 30, 2 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atoms (referred to herein as C2 to C6, respectively). 30 Alkenyl, C2-C 18 Alkenyl, C 14 ~C 24 Alkenyl, C 14 ~C 18 Alkenyl, C 16 Alkenyl, or C 18 It may also refer to a functional group having an oxygen atom linking an unsaturated straight chain or branched hydrocarbon, such as an alkyl group (alkenyl), to a second unsaturated straight chain or branched hydrocarbon.

[0023] As used herein, the term "vinyl ether" refers to a group in which the alkene is adjacent to an ether linkage.

[0024] The term "ester" as used herein refers to any organic compound having a structure similar to an ester functional group. As used herein, the term "ester" refers to any compound or functional group that can be represented by the general formula -RCOOR', where R is hydrogen or a straight or branched chain group of 1 to 30, 1 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atoms (referred to herein as C1 to C5, respectively). 30 Alkyl, C1-C 18 Alkyl, C 14 ~C 24 Alkyl, C 14 ~C 18 Alkyl, C 16 Alkyl or C 18 alkyl), or straight or branched groups of 2 to 30, 2 to 18, 14 to 24, 14 to 18, 16 or 18 carbon atoms (referred to herein as C2 to C6, respectively). 30 Alkenyl, C2-C 18 Alkenyl, C 14 ~C 24 Alkenyl, C 14 ~C 18 Alkenyl, C 16 Alkenyl, or C 18 R' is a saturated or unsaturated straight chain or branched hydrocarbon group such as 1 to 30, 1 to 18, 14 to 24, 14 to 18, 16, or 18 carbon atom straight chain or branched group (referred to herein as C1 to C6, respectively). 30 Alkyl, C1-C 18 Alkyl, C 14 ~C 24 Alkyl, C 14 ~C 18 Alkyl, C 16 Alkyl or C 18 alkyl), or straight or branched groups of 2 to 30, 2 to 18, 14 to 24, 14 to 18, 16 or 18 carbon atoms (referred to herein as C2 to C6, respectively). 30 Alkenyl, C2-C18 Alkenyl, C 14 ~C 24 Alkenyl, C 14 ~C 18 Alkenyl, C 16 Alkenyl, or C 18 The alkyl groups are saturated or unsaturated, straight chain or branched hydrocarbons, such as alkyl groups (also called alkenyl groups).

[0025] The terms "substituted" and "optionally substituted" with respect to a phosphate group, an alkyl group, an alkenyl group, or an acyl group mean that the group is optionally substituted with an additional moiety. In each case, the substituted or additional moiety may be hydrogen, C 1-3 It can be independently selected from alkyl, and halogen (e.g., Cl, F, Br, or I). In certain embodiments, the group is not substituted, i.e., unsubstituted.

[0026] The lipid nomenclature used here follows the guidelines established by the Lipid Maps Consortium and the abbreviations of Liebisch et al. (Liebisch et al., Fahy et al. (2009), Fahy et al. (2005)). Alternatively, the nomenclature is based on "Nomenclature of Lipids" in Biochemical Nomenclature and Related Documents, 2nd ed., Portland Press (London, 1992), edited by C. Liebecq, ISBN 1-85578-005-4. Phospholipids typically contain two fatty acid chains and, in the absence of detailed characterization, are expressed as the total composition of carbon atoms and double bonds (e.g., PC(38:6)). However, if the acyl chain composition has been determined, the nomenclature will indicate this (e.g., PC(38:6) is changed to PC(16:0 / 22:6)).

[0027] The present disclosure relates to lipid molecules that use the numbering system X:Y, where the number X represents the number of carbon atoms present in the chain.

[0028] In the context of alkylglycerols, alkylacylglycerols, and alkyldiacylglycerols, the number Y represents the number of double bonds present in the chain. For example, an alkylglycerol numbered 16:0 contains a hydrocarbon group with a 16-carbon chain and no double bonds. As a further example, an alkylglycerol numbered 18:1 contains a hydrocarbon group with an 18-carbon chain and one double bond.

[0029] In the context of plasmalogens / plasmenyl phospholipids, the number Y in the first listed alkenyl chain (i.e., PE(PX:Y / X:Y)) represents the number of double bonds present in the alkenyl chain in addition to the vinyl ether group. For example, in plasmalogens numbered PE(P-16:0 / 20:4), the 16:0 alkenyl group contains a hydrocarbon group with 16 carbon atoms and no double bonds other than the vinyl ether group (i.e., a double bond exists between the first two carbon atoms, and the remaining 14 carbon atoms are saturated). As another example, in plasmalogens numbered PE(P-18:1 / 20:4), the 18:1 alkenyl group contains a hydrocarbon group with an 18-carbon chain with one double bond in addition to the vinyl ether group (i.e., a double bond exists between the first two carbon atoms, and one other double bond exists between two of the remaining 16 carbon atoms). As shown in Scheme I above, plasmenyl PE plasmalogen is represented by the structure PE(P-16:0 / 22:6).

[0030] When an ether lipid contains one or more double bonds, the double bonds may be at various positions in the hydrocarbon chain. For example, an alkylglycerol numbered 18:1 may contain a mixture of species with cis-n1 and cis-n9 double bonds. As another example, a plasmalogen numbered 18:1 (e.g., PE(P)) may contain a mixture of species with cis-n1 and cis-n9 double bonds.

[0031] As used herein, the term "plasmanyl" is understood to refer to a phospholipid having an ether bond at the sn-1 position to the alkyl group.

[0032] As used herein, the term "plasmenyl" is understood to refer to a phospholipid having an ether bond at the sn-1 position relative to an alkenyl group. Plasmenyl phospholipids are also called "plasmalogens."

[0033] Plasmalogens with "15:0" alkenyl groups are molecules that contain a double bond (typically a cis-vinyl ether group) between carbon 1 and carbon 2 and an ether bond at the sn-1 position to a 15-carbon chain that contains no other double bonds in the chain.

[0034] Plasmalogens with "16:0" alkenyl groups are molecules that contain a double bond (typically a cis-vinyl ether group) between carbon 1 and carbon 2 and an ether bond at the sn-1 position to a 16-carbon chain that contains no other double bonds in the chain.

[0035] Plasmalogens with "17:0" alkenyl groups are molecules that contain a double bond (typically a cis-vinyl ether group) between carbon 1 and carbon 2 and an ether bond at the sn-1 position to a 17-carbon chain that contains no other double bonds in the chain.

[0036] Plasmalogens with "18:0" alkenyl groups are molecules that contain a double bond (typically a cis-vinyl ether group) between carbon 1 and carbon 2 and an ether bond at the sn-1 position to an 18-carbon chain that contains no other double bonds in the chain.

[0037] Plasmalogens with "18:1" alkenyl groups are molecules that contain a double bond (typically a cis-vinyl ether group) between carbons 1 and 2, typically one additional double bond between carbons 7 and 8 (e.g., n7), between carbons 9 and 10 (e.g., n9), or between carbons 11 and 12 (e.g., n11), and typically have an ether bond at the sn-1 position relative to an 18-carbon chain with a cis double bond.

[0038] Plasmalogens with "20:0" alkenyl groups are molecules that contain a double bond (typically a cis-vinyl ether group) between carbon 1 and carbon 2 and an ether bond at the sn-1 position to a 20-carbon chain that contains no other double bonds in the chain.

[0039] Plasmalogens with "20:1" alkenyl groups are molecules containing a double bond (typically a cis-vinyl ether group) between carbons 1 and 2, typically one additional double bond between carbons 7 and 8 or between carbons 9 and 10, and typically an ether bond at the sn-1 position to a 20-carbon chain with a cis-double bond.

[0040] Plasmalogens with "18:2" acylalkenyl groups are molecules with an ester bond at the sn-2 position to an 18-carbon chain, typically with two double bonds between carbons 9 and 10 and between carbons 11 and 12, and typically with a cis-double bond.

[0041] Plasmalogens with a "20:4" acylalkenyl group are molecules with four double bonds, typically between carbons 5 and 6, between carbons 8 and 9, between carbons 11 and 12, and between carbons 14 and 15, and an ester bond at the sn-2 position of a 20-carbon chain with a cis-double bond.

[0042] Plasmalogens with a "22:6" acylalkenyl group are molecules with six double bonds, typically between carbons 4 and 5, between carbons 7 and 8, between carbons 10 and 11, between carbons 13 and 14, between carbons 16 and 17, and between carbons 19 and 20, and an ester bond at the sn-2 position to a 22-carbon chain with a cis-double bond.

[0043] As used herein, "acylalkenyl" refers to a straight or branched chain hydrocarbon, e.g., containing 2 to 30 carbons and containing at least one carbon-carbon double bond covalently linked to an acyl group. For example, the use of nomenclature such as 22:6 or 18:2 in the context of acylalkenyl groups refers to acylalkenyl groups having 22 carbons or 18 carbons, respectively, and 6 or 2 double bonds, respectively. Examples of acylalkenyl groups are: [ka]

[0044] Acylalkenyl groups can occur (as acyl groups) in species such as alkylacylglycerols or alkyldiacylglycerols, or as acyl groups in plasmanyl-phospholipids or plasmenyl-phospholipids. Typically, when present in these species, there is no double bond between the carbons α- and β-to the acyl group.

[0045] As used herein, "acylalkyl" refers to a straight or branched chain hydrocarbon, e.g., containing 1 to 30 carbons, covalently attached to an acyl group. For example, the use of the nomenclature 22:0 or 18:0 in the context of acylalkyl groups refers to acylalkyl groups having 22 carbons or 18 carbons, respectively. Examples of acylalkyl groups are: [ka]

[0046] It will also be recognized that the compounds described herein may have asymmetric centers and, therefore, can exist in multiple stereoisomeric forms. Accordingly, the present disclosure also relates, at least in some embodiments, to compounds that are substantially pure isomeric at one or more asymmetric centers, e.g., having an enantiomeric purity of 90% enantiomeric excess ("ee") or greater, e.g., 95% ee, 97% ee, 99% ee, or greater than 99% ee. The present disclosure also relates to compounds that exist as mixtures of stereoisomeric forms, including racemic, diastereomeric, or scalemic mixtures. Such isomers may be naturally occurring or may be prepared by asymmetric synthesis, including, but not limited to, synthetic methods using chiral intermediates, or by chiral resolution. Certain compounds contained in the compositions of the present disclosure may exist in particular geometric or stereoisomeric forms. The present disclosure contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are intended to be included. For example, if a specific enantiomer of a compound of the present disclosure is desired, the enantiomer may be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary; the resulting diastereomeric mixture is separated; and the auxiliary is cleaved to provide the pure desired enantiomer. Alternatively, if the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts can be formed with an appropriate optically active acid or base, followed by resolution of the formed diastereoisomers by fractional crystallization or chromatographic means well known in the art, followed by recovery of the pure enantiomers.

[0047] The present disclosure also relates to derivatives of glycerol. While glycerol is achiral, the derivatives are typically chiral. Typically, the glycerol utilized has a stereochemical configuration corresponding to that found in nature. In some embodiments, the glycerol derivative utilized has the stereochemical configuration shown above in formula (I).

[0048] As referred to herein, the term "alkylglycerol" refers to R 1 The group is a hydrocarbon chain, and R 2 Groups and R 3 The term "alkylglycerol" is further understood to mean a compound of formula (IA), wherein R A is the hydrocarbon chain and R x Each group is independently hydrogen or C 1-3 The term "alkylglycerol" is further intended to mean compounds of formula (I-A1), (I-A2) and (I-A3). Although the term "alkylglycerol" is used, this term does not include alkyl groups containing unsaturation in the hydrocarbon chain, such as R 1 Those skilled in the art will understand that "alkylglycerol" compounds encompass species having hydrocarbon groups at positions 1 and 2 of the hydrocarbon chain. However, as used herein, "alkylglycerol" compounds do not contain a double bond between carbons 1 and 2 of the hydrocarbon chain, i.e., proximal to the ether linkage.

[0049] Alkylglycerols with a "16:0" group are molecules with an ether bond at the sn-1 position to a saturated hydrocarbon chain of 16 carbon atoms and no double bonds in the chain, and may also be known as chimyl alcohols.

[0050] Alkylglycerols with an "18:0" group are molecules with an ether bond at the sn-1 position to an 18-carbon saturated hydrocarbon chain and no double bonds in the chain, and may also be known as batyl alcohols.

[0051] Alkylglycerols with an "18:1" group are molecules with an ether bond at the sn-1 position to an 18-carbon saturated hydrocarbon chain that typically contains one double bond, typically a cis double bond, between carbons 9 and 10, and may also be known as selachyl alcohols.

[0052] As referred to herein, the term "alkyl acylglycerol" means R 1 The group is a hydrocarbon chain, and R 2 Groups and R 3 One of the groups is hydrogen and R 2 Groups and R 3 The term "alkyl acyl glycerol" refers to a compound of formula (I) in which the other of the groups is an acyl group independently selected from acyl alkyl and acyl alkenyl groups. Although the term "alkyl acyl glycerol" is used, this term does not include alkyl acyl glycerols containing unsaturation in the hydrocarbon chain. 1 It will be understood by those skilled in the art that the term "alkyl acylglycerol" encompasses species having hydrocarbon groups at the R 1 The hydrocarbon chain does not contain a double bond between carbon 1 and carbon 2, i.e., proximal to the ether bond.

[0053] As referred to herein, the term "alkyl diacylglycerol" means R 1 The group is a hydrocarbon chain, and R 2 Groups and R 3 The term "alkyl diacylglycerol" refers to a compound of formula (I) where the R groups are acyl groups independently selected from acyl alkyl and acyl alkenyl. Although the term "alkyl diacylglycerol" is used, this term does not include alkyl diacylglycerols containing unsaturation in the hydrocarbon chain. 1 It will be understood by those skilled in the art that the term "alkyl diacylglycerol" encompasses species having hydrocarbon groups at the R 1 The hydrocarbon chain does not contain a double bond between carbon 1 and carbon 2, i.e., proximal to the ether bond.

[0054] As used herein, the term "LPX(O)" refers to LPC(O) or LPE(O) as used elsewhere herein. In some embodiments, LPX(O) is selected from one or more of LysoPAF-C and LysoPAF-O. In some embodiments, LysoPAF-C is selected from one or more of LysoPAF-C16, LysoPAF-C18, and LysoPAF-C18:1.

[0055] The term "fatty acid" may refer to, for example, stearic acid, palmitic acid, or oleic acid.

[0056] As used herein, the terms "subject" and "patient" refer to a human or animal treated by the methods of the present disclosure. In one embodiment, the subject is a human. In one embodiment, the subject is an animal, such as a domestic animal, a working animal, or a farm animal. Domestic animals include, but are not limited to, rabbits, birds, cats, dogs, fish, rats, turtles, reptiles (lizards, snakes), and the like. Working animals include, but are not limited to, cattle, yaks, and horses. Farm animals include, but are not limited to, sheep, pigs, cows, chickens, goats, geese, ducks, llamas, and the like. In some embodiments, it is contemplated that the compositions described herein are formulated as animal feed or animal nutritional supplements. In some embodiments, administration may be oral. In some embodiments, administration may be by any suitable route, including liquid, capsule, tablet, or lozenge.

[0057] As used herein, the term "non-disease state" refers to a state in which the subject is not suffering from a plasmalogen-related disease or deficiency that requires treatment.

[0058] As used herein, the term "treating" includes any effect, e.g., alleviating, reducing, increasing, maintaining, modulating, improving, or eliminating, that results in the improvement of a condition, disease, disorder, etc., or that ameliorates the symptoms thereof.

[0059] As used herein, the term "effective amount" refers to an amount of a compound (e.g., a compound of the present disclosure) sufficient to produce a beneficial or desired result. An effective amount can be administered in one or more administrations, applications, or dosages, and is not intended to be limited to a particular formulation or route of administration.

[0060] As used herein, the term "composition" refers to a product containing specified ingredients in specific amounts, and any product resulting directly or indirectly from the combination of specified ingredients in specific amounts. In some embodiments, a composition includes an active ingredient and an inactive ingredient. In some embodiments, a composition is a formulation. In some embodiments, a formulation is a composition suitable for administration to and / or ingestion by a subject, such as a human. A composition may be a pharmaceutical composition, and a formulation may be a pharmaceutical formulation. In some embodiments, a pharmaceutical composition or pharmaceutical formulation includes a combination of an active agent and a pharmaceutically acceptable carrier, diluent, excipient, solubilizer, or vehicle (inactive or active). As used herein, the term "pharmaceutically acceptable" means that the carrier, diluent, excipient, solubilizer, or vehicle is compatible with the other ingredients of the formulation and non-toxic to the subject. Thus, it is understood that a "pharmaceutical composition" or a "pharmaceutical formulation" is suitable for administration to and / or ingestion by a subject, such as a human, and may be approved for such administration and / or ingestion by, for example, the U.S. Food and Drug Administration and / or the European Medicines Agency. It should also be understood that compositions or formulations may not necessarily be approved, for example, by the U.S. Food and Drug Administration and / or the European Medicines Agency, for administration to and / or consumption by subjects, such as humans.

[0061] Pharmaceutical compositions include products containing an active ingredient and an inactive ingredient that constitutes a carrier, and include any product that results directly or indirectly from the combination, complexation or aggregation of any two or more ingredients, or the dissociation, or other type of reaction or interaction of one or more ingredients. Thus, pharmaceutical compositions of the present disclosure include any composition prepared by mixing at least one compound of the present disclosure with a pharmaceutically acceptable carrier, diluent, excipient, solubilizer, or vehicle.

[0062] In some embodiments, the formulations of the present disclosure are in the form of a beverage or food. In some embodiments, the beverage or food is formulated for public consumption, such as food grade. In some embodiments, the beverage or food is formulated as a dietary supplement or other nutritional composition. In some embodiments, the beverage or food is pharmaceutical grade. For the avoidance of doubt, it is understood that in some embodiments, the formulations of the present disclosure may be suitable for consumption by a subject, such as a human, but need not necessarily be pharmaceutical grade (e.g., food grade or dietary supplement grade, but not necessarily pharmaceutical grade).

[0063] In some embodiments, the formulation may contain one or more of a solubilizer, emulsifier, stabilizer, dispersant, antifoaming agent, or diluent. The formulation may further contain one or more antioxidant compounds. It is anticipated that any antioxidant suitable for oral administration, such as vitamin A, vitamin E, vitamin C, retinal, tocopherol, L-ascorbyl palmitate, riboflavin, and carotenoids, including lutein, beta-carotene, zeaxanthin, and lycopene, and combinations thereof, may be formulated into the embodiments described herein. In some embodiments, the composition is inherently insensitive to oxidation and does not require the addition of antioxidant compounds.

[0064] As used herein, the term "excipient" is intended to mean an inactive ingredient used as a vehicle (e.g., water, capsule shell, etc.), diluent, or component of a dosage form or pharmaceutical composition containing a drug, such as a therapeutic agent. The term also includes inactive ingredients that impart cohesive functions (e.g., binders), disintegration functions (e.g., disintegrants), lubricating functions (e.g., lubricants), and / or other functions (e.g., solvents, surfactants, etc.) to the composition.

[0065] As used herein, the term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt (e.g., acid or base) of a compound of the present disclosure, which, when administered to a subject, can provide a compound of the present disclosure or its active metabolite or residue. For example, Berge et al. provide a detailed description of pharmaceutically acceptable salts in J. Pharmaceutical Sciences (1977) 66:1-19. As known to those skilled in the art, "salts" of compounds of the present disclosure can be derived from inorganic or organic acids and bases. Salts can be prepared in situ during the final isolation and purification of compounds of the present disclosure, or separately by reacting a free base function with a suitable organic acid. Examples of acids include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, ethanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Other acids, while not themselves pharmaceutically acceptable, may be used in the preparation of salts useful as intermediates to obtain the compounds of the present disclosure and their pharmaceutically acceptable acid addition salts. Examples of bases include alkali metal (e.g., sodium) hydroxides, alkaline earth metal (e.g., magnesium) hydroxides, ammonia, and bases of the formula NW4 + wherein each W is H or C 1-4alkyl). Basic nitrogen-containing groups can be quaternized with agents such as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and diamyl sulfate; long chain alkyl halides, such as decyl, lauryl, myristyl, and stearyl; chlorides, bromides, and iodides; aryl alkyl halides, such as benzyl bromide and phenethyl bromide, and others. This can result in products with improved solubility and dispersibility.

[0066] Examples of salts include, but are not limited to, acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, palmitate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, etc. Other examples of salts include those containing an appropriate cation, e.g., Na + , NH4 + , and NW4 + (where each W is H or C 1-4 Included are anions of the compounds of the present disclosure combined with alkyl, alkyl groups, and the like. For therapeutic use, salts of the compounds of the present disclosure are contemplated as being pharmaceutically acceptable. However, salts of acids and bases that are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

[0067] As used herein, the term "pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers, such as phosphate buffered saline, water, emulsions (e.g., oil / water or water / oil emulsions), and various types of wetting agents. The composition may also contain stabilizers and preservatives. For examples of carriers, stabilizers, and adjuvants, see Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA).

[0068] As used herein, in some embodiments, the term "carrier" refers to a pharmaceutically acceptable carrier. In some embodiments, the term "carrier" refers to any suitable carrier, which may not be pharmaceutical grade. For example, in some embodiments, the carrier may be food grade, but not pharmaceutical grade.

[0069] As used herein, the term "day" is intended to mean a 24-hour period.

[0070] As a general matter, compositions expressing percentages are by weight unless otherwise specified.

[0071] Ether lipids and compositions The present disclosure demonstrates that ether lipids, such as plasmanyl-phospholipids and plasmenyl-phospholipids (plasmalogens), have in vivo profiles that are associated with health and pathological conditions such as diabetes. It has also been found that the in vivo ether lipid profile can be influenced by administering to a subject a composition comprising the ether lipids, e.g., by supplementing with krill oil.

[0072] The formulations of the present disclosure are useful for maintaining and / or altering in vivo ether-lipid levels to levels and / or ratios associated with a natural, non-disease state (i.e., the natural state of a healthy human subject) and / or for altering or modulating in vivo ether-lipid levels toward levels and / or ratios associated with a natural, non-disease state (i.e., the natural state of a healthy human subject). An illustrative, but non-limiting example, of the term "healthy human subject" is taken to mean a subject or patient not suffering from a disease or disorder associated with plasmalogen deficiency.

[0073] Examples of compounds useful for maintaining and / or modifying in vivo ether lipid levels and / or their ratios include alkylglycerols, alkylacylglycerols (compounds that are ethers derived from glycerol alcohol and alkyl alcohol, with an acyl group derived from another glycerol alcohol and an acid), alkyldiacylglycerols, and ether phospholipids, including, but not limited to, LPC(O), LPE(O), LPA(O), plasmanyl-phospholipids, and plasmenyl-phospholipids. In some embodiments, the formulations are for maintaining and / or modifying in vivo plasmanyl-phospholipid and / or plasmenyl-phospholipid levels and / or their ratios. In some embodiments, the plasmanyl-phospholipid and / or plasmenyl-phospholipid include those having a phosphatidylcholine group and / or a phosphatidylethanolamine group. In some embodiments, the formulations are for maintaining and / or modifying in vivo plasmenyl-phospholipid (plasmalogen) levels and / or their ratios. In some embodiments, the formulation is for the in vivo maintenance and / or alteration of phosphatidylethanolamine plasmenyl-phospholipid (plasmalogen) levels and / or ratios thereof. [Table 2]

[0074] As described herein, healthy subjects tend to have plasmanyl-phospholipid and plasmenyl-phospholipid profiles in which specific alkenyl ether and alkyl ether groups are present. For example, a group of healthy subjects was found to have a high proportion of ether lipids (e.g., plasmanyl-phospholipids and / or plasmenyl-phospholipids containing plasmalogens) with 18:1 alkenyl ether groups, 18:0 alkyl ether groups, and 16:0 alkyl ether groups. Note that the listed alkyl / alkenyl groups also contain a double bond (cis-vinyl ether group) between carbon 1 and carbon 2, as described above for some exemplary plasmalogen compounds.

[0075] In some embodiments, the formulations are for the in vivo maintenance and / or alteration of ether lipids having 18:1 alkenyl ether groups, 18:0 alkyl ether groups, and 16:0 alkyl ether groups (e.g., plasmanyl-phospholipids and / or plasmenyl-phospholipids). In some embodiments, the formulations are for the in vivo maintenance and / or alteration of levels of plasmalogens having 18:1 ether groups, 18:0 ether groups, and 16:0 ether groups. In some embodiments, the formulation is for in vivo maintenance of an ether lipid (e.g., plasmanyl-phospholipid and / or plasmenyl-phospholipid) in a total ether lipid (e.g., plasmanyl-phospholipid and / or plasmenyl-phospholipid) profile in vivo, or for in vivo modification of an ether lipid (e.g., plasmanyl-phospholipid and / or plasmenyl-phospholipid) in a total ether lipid (e.g., plasmanyl-phospholipid and / or plasmenyl-phospholipid) profile in vivo, wherein the ether lipid (e.g., plasmanyl-phospholipid and / or plasmenyl-phospholipid) has a molar ratio of 18:1 alkenyl ether groups to 18:0 alkyl ether groups to 16:0 alkyl ether groups of about 1:1.7:1.4.In some embodiments, the formulation is for in vivo maintenance of ether lipids (e.g., plasmanyl-phospholipids and / or plasmenyl-phospholipids) in a total ether lipid (e.g., plasmanyl-phospholipids and / or plasmenyl-phospholipids) profile in vivo, or for in vivo alteration of ether lipids (e.g., plasmanyl-phospholipids and / or plasmenyl-phospholipids) in a total ether lipid (e.g., plasmanyl-phospholipids and / or plasmenyl-phospholipids) profile in vivo, wherein the total ether lipid (e.g., plasmanyl-phospholipids and / or plasmenyl-phospholipids) in vivo In the plasmenyl-phospholipid) profile, the ether lipid (e.g., plasmenyl-phospholipid and / or plasmenyl-phospholipid) has a molar percentage of 18:1 alkenyl ether groups in the range of 18.6% to 27.9%, a molar percentage of 18:0 alkyl ether groups in the range of 32.6% to 45.8%, and a molar percentage of 16:0 alkyl ether groups in the range of 26.8% to 37.4%, or a molar percentage of 18:1 alkenyl ether groups of about 23.3%, a molar percentage of 18:0 alkyl ether groups of about 39.2%, and a molar percentage of 16:0 alkyl ether groups of about 32.1%.

[0076] In some embodiments, the formulation is for in vivo maintenance of an ether lipid in an in vivo plasmalogen lipid profile or for in vivo alteration of an ether lipid to an in vivo plasmalogen lipid profile, wherein the ether lipid has a molar ratio of 18:1 ether groups to 18:0 ether groups to 16:0 ether groups of about 1:1.7:1.4. In some embodiments, the formulation is for in vivo maintenance of an ether lipid at, or in vivo alteration of, an in vivo plasmalogen lipid profile, wherein the ether lipid has a molar percentage of 18:1 ether groups in the range of 18.6% to 27.9%, a molar percentage of 18:0 ether groups in the range of 32.6% to 45.8%, and a molar percentage of 16:0 ether groups in the range of 26.8% to 37.4%, or a molar percentage of 18:1 ether groups of about 23.3%, a molar percentage of 18:0 ether groups of about 39.2%, and a molar percentage of 16:0 ether groups of about 32.1%.

[0077] In some embodiments, the composition comprises at least one isolated compound. In some embodiments, the at least one isolated compound is found to be greater than 99% pure. In preferred embodiments, the at least one isolated compound is found to be greater than 99.9% pure.

[0078] In some embodiments, the composition comprises a mixture of at least two compounds, where the % (w / v) of one compound is at least 90%.

[0079] In one aspect, dietary plasmalogens have been studied after krill oil (KO) supplementation in humans, as described in Sung, et al., "Enrichment of n-3 containing ether phospholipids in plasma after 30 days of krill oil compared with fish oil supplementation," Lipids (2022) 57:115-124, the entire contents of which are incorporated herein by reference. It is recognized that ether-containing phospholipids are minor components in plasma and include either alkyl or alkenyl phosphatidylcholines and phosphatidylethanolamines (the latter alkenyl class is referred to as plasmalogens). Furthermore, it should be noted that although plasmalogen levels are lower than diacyl lipids, plasmalogens are still highly abundant. These have been studied more recently as techniques such as lipidomics have enabled the detection and quantification of these and other minor lipid classes (see, e.g., Meikle, et al., “Postprandial plasma phospholipids in men are influenced by the source of dietary fat,” J. Nutr. (2015) 145(9):2012-2018). Methods for making certain ether lipid and / or plasmalogen compositions are described in WO 2021 / 007623 A1, Baker Heart and Diabetes Institute, inventor Meikle, which is incorporated herein by reference in its entirety. There are numerous approaches to synthesizing these ether lipid compounds, using either exclusively chemical synthesis or a combination of chemical and enzymatic synthetic strategies.The synthesis of AKG compounds has been reported in the following papers: Carlos D. Magnusson, et al., "Chemoenzymatic synthesis of a focused library of enantiopure structured 1-O-alkyl-2,3-diacyl-sn-glycerol type ether lipids," Tetrahedron (2011) 67: 1821-1836; and Arnar Halldorsson, et al., "Lipase catalyzed kinetic resolution of 1-O-alkylglycerols by sequential transesterification," Tetrahedron: Asymmetry (2004) 15: 2893-2899. Similarly, the synthesis of lysoalkenylphosphatidylethanolamine (LPE(P)) from commercially available 2,3-O-isopropylidene-sn-glycerol is described in Guanghui Ni, et al., "Synthesis and evaluation of immunostimulant plasmalogen lysophosphatidylethanolamine and analogues for natural killer T cells," Bioorg. Med. Chem. (2014) 22(11):2966-73. Gomes, MAGB, et al. describe the synthesis of a number of alkyl ether lipids (Gomes MAGB, Bauduin A, Le Roux C, Fouinneteau R, Berthe W, Berchel M, Couthon H, Jaffres PA. "Synthesis of ether lipids: natural compounds and analogues," Beilstein J Org Chem. 2023 Sept. 8;19:1299-1369). Figure 2 of the Gomes reference discusses a synthetic route that can be adapted to produce the desired alkyl LPC(O) compounds, providing the R group of the desired sn-1 fatty acid and terminating in the lyso PAF step. One skilled in the art will recognize that different protecting groups and / or deprotection schemes can also be used.Figure 6 of Gomes and further references disclosed therein describe methods for producing the starting compound 1-O-alkylglycerol (i.e., compound 2.1 in Figure 2 of Gomes). Additionally, U.S. Patent No. 10,900,063, the entire contents of which are incorporated herein by reference, describes the use of lipases to produce LPC(O).

[0080] LPC(O) synthesis pathway [ka] Scheme II. Synthesis of C18.0 LPC(O)

[0081] Starting material 1 (R-solketal or R-(-)-2,3-O-isopropylidene-sn-glycerol; CAS 14347-78-5; available from CombiBlocks (98% purity) or Fluorochem (95% purity)) is condensed with either an alkyl bromide or mesylate to form the corresponding alkyl-substituted acetonide (compound 2). TBAB, shown in Scheme II above, is tetrabutylammonium bromide, a commonly used phase transfer catalyst. The isopropylidene group can then be removed by acid hydrolysis (compound 3), and an alternative protecting group can be applied to give compound 6. Separately, 2-bromoethyl phosphorodichloridate (compound 8) is prepared and then condensed with compound 6 to give compound 9. The terminal bromine atom is then replaced with trimethylamine in a final alcohol deprotection step to give compound 11.

[0082] As will be readily apparent to those skilled in the art, the reagents and conditions proposed above are merely illustrative and not limiting. The synthesis of other SN-1 substituted LCP(O)s can be achieved, for example, by providing the appropriate alkyl bromide or alkyl mesylate compound in Step 1. For example, 1-bromooctadecane (CAS 112-89-0, available from TCI Chemicals, purity greater than 97.0% as determined by gas chromatography), oleyl mesylate (prepared by mesylation of oleyl alcohol: CAS 143-28-2, available from Sigma-Aldrich, purity 85% as determined by gas chromatography), and 1-bromohexadecane (CAS 112-82-3, available from TCI Chemicals, purity 96% as determined by gas chromatography) can be used to synthesize C18:0, C18:1, and C16:0 LCP(O), respectively.

[0083] Suitable protecting groups for alcohol moieties are well known in the art (see, for example, Greene's Protective Groups in Organic Synthesis, Wiley, DOI:10.1002 / 9781118905074).

[0084] LPE(O) synthesis route The synthesis of the corresponding LPE(O) compound can be carried out in a similar manner using a substituted ammonium compound in place of trimethylamine in step 8. The ammonium compound is then deprotected and reacted with NH + Such deprotection may occur in conjunction with the alcohol deprotection in step 9 or via an additional synthetic step, depending on the nature of the selected substituents and protecting groups. An alternative exemplary, non-limiting synthesis is: (i) saturating the vinyl double bond of the LPE(P) alkenyl compound produced by the method of the Ni et al. reference by chemical (typically nickel) or enzymatic reduction to obtain an alternative synthesis of LPE(O); and (ii) Commercially available alkyl PE compounds, such as Avanti Polar Lipids (<avantilipids.com> ) by chemical or enzymatic means, for example using an appropriate lipase, to give C16.O alkyl LPE(O).

[0085] In certain embodiments, combinations of lysoalkylphosphatidylcholine (LPC(O)) or lysoalkylphosphatidylethanolamine (LPE(O)) compounds are contemplated. Derivatives of the foregoing compounds are also contemplated. One exemplary embodiment includes a specific formulation of three species of lysoalkylphosphatidylcholine (LPC(O)). These species may include LPC(O-18:1), designated below as compound (I-A1), LPC(O-18:0), designated below as compound (I-A2), and LPC(O-16:0), designated below as compound (I-A3). These ether lipids may be R 1 The molar ratio of the groups may be, for example, about 1.0:1.7:1.4. These ratios may vary between different cell target populations and different subjects.

[0086] In this preferred embodiment, the compound is shown as follows, as LPC(O-18:1): [ka]

[0087] As LPC(O-18:0): [ka]

[0088] As LPC(O-16:0): [ka]

[0089] Other compounds and combinations are also contemplated, such as lysoalkylphosphatidylethanolamine (LPE(O)) combinations including LPE(O-18:1), shown below as compound (I-A4), LPE(O-18:0), shown below as compound (I-A5), and LPE(O-16:0), shown below as compound (I-A6). 1 The molar ratios of groups, as well as mole percentages and weight percentages, can vary.

[0090] In this embodiment, the compound is shown as follows, as LPE(O-18:1): [ka]

[0091] As LPE(O-18:0): [ka]

[0092] As LPE(O-16:0): [ka]

[0093] Other compounds and combinations are also contemplated, such as the combination of lysoalkylphosphatidic acid (LPA(O)), a phosphate compound containing LPA(O-18:1), shown below as compound (I-A7), LPA(O-18:0), shown below as compound (I-A8), and LPA(O-16:0), shown below as compound (I-A9). 1 The molar ratios of groups, as well as mole percentages and weight percentages, can vary.

[0094] In this embodiment, the compound is shown as follows, as LPA(O-18:1): [ka]

[0095] As LPA(O-18:0): [ka]

[0096] As LPA(O-16:0): [ka]

[0097] Upon ingestion, these phosphocholine species, particularly LPC(O), are absorbed in the intestine and metabolized into a series of ether phospholipids, including alkylphosphatidylcholine (PC(O)), alkylphosphatidylethanolamine (PE(O)), alkenylphosphatidylcholine (plasmalogen, PC(P)), and alkenylphosphatidylethanolamine (plasmalogen, PE(P)). Studies have shown that this results in increased levels of both PC(P) and PE(P) species, bioactive species with several desirable health properties. The bioavailability of LPC(O) and its conversion to PC(P) and PE(P) is 5-12 times higher than that of alkylglycerol (AKG) species, which are also commonly used to increase plasmalogen levels. Without wishing to be bound by theory, this is thought to mean that a lower dose can be used to achieve the same increase in plasmalogen levels.

[0098] A further embodiment contemplated by the present disclosure is a formulation, which may be in the form of a food product such as a dietary supplement, that, upon ingestion, elevates plasmalogen levels in the recipient's blood and tissues, thereby improving or overcoming any plasmalogen deficiency that may exist in the individual. Increased plasmalogen levels may lead to improved health outcomes. Such formulations, for example, LPC(O) formulations, may also be incorporated into a range of foods to facilitate delivery to the recipient. The intended recipient may be anyone who is plasmalogen deficient and / or at risk for any of a variety of metabolic diseases for which plasmalogens may play a protective role.

[0099] As used herein, the term "dietary supplement" refers to a food product intended to enhance a subject's diet, thereby improving nutrition. Dietary supplements may include the compositions described herein, alone or in combination with other ingredients intended to supplement the diet, such as vitamins and minerals, dietary fiber, herbal and other botanical extracts (including flower remedies, homeopathic remedies, amino acids, enzymes and live microorganisms, probiotics, prebiotics, or any combination thereof). Dietary supplements may be formulated in a variety of ways, including as oils, gummies, drops, capsules, fast-dissolving formulations, lozenges, oral sprays, chewing gum, gels, powders, premixed drinks, meal replacement shakes, and bars.

[0100] An example of a food product that can incorporate the formulations described herein is infant formula. Food products that can incorporate formulations containing, for example, LPC(O) include infant formula, follow-on formula, medical foods, and foods for special medical purposes.

[0101] An example of a food product into which the compositions or formulations described herein can be incorporated is any food product formulated for human consumption.

[0102] As used herein, the term "medical food" or "food for special medical purposes" refers to a food specially formulated and intended for the dietary management of a disease, disorder, or condition with unique nutritional needs that cannot be met through regular diet alone. Medical foods assist patients who are temporarily or permanently unable to achieve adequate nutritional intake from regular foods or by modifying their regular diet and who are malnourished or at risk of becoming malnourished. Medical foods are used under medical supervision and can be administered orally or via tube feeding (e.g., nasogastric tube). As used herein, these terms are referred to under Regulation (EU) No. 609 / 2013 and (Food and Drug Administration (FDA)) 21 CFR 101.9(j)(8)(ii). They are therefore distinguished from dietary supplements, which are generally available for consumption without medical supervision.

[0103] The term "infant" as used herein is intended to mean a person 12 months of age or younger. The term "infant" is further intended to mean a "premature infant" as a person 12 months of age or younger born before 36 weeks of gestation. The term "toddler" as used herein is intended to mean a person between 1 and 3 years of age. The terms "child" or "children" as used herein refer to a person between 3 and 12 years of age. The term "infant formula" as used herein, unless otherwise specified, refers to liquid, semi-liquid, solid, and semi-solid human breast milk substitutes or substitutes suitable for consumption by infants. Synthetic infant formula includes ingredients of semi-purified or purified origin. As used herein, the terms "semi-purified" and "purified" refer to substances prepared by purification of natural substances or synthesis. The term "infant formula" is not intended to include unaltered human breast milk.

[0104] Infant formulas may include liquid and powder nutritional supplements, liquid and powder human breast milk fortifiers, liquid and powder premature infant formulas, liquid and powder infant formulas, liquid and powder elemental and semi-elemental formulas, liquid and powder toddler formulas, and powder follow-on formulas suitable for use by infants and children. The compositions may be in any product form that contains the ingredients described herein and is safe and effective for oral administration.

[0105] Infant formulas may further include ingredients including protein, fat, carbohydrates, vitamins, minerals, anti-caking agents, and emulsifiers. In some embodiments, the formula may include purified cow's milk whey and / or casein as a protein source, a vegetable oil blend as a fat source, lactose as a carbohydrate source, a vitamin-mineral mixture, and other ingredients including, but not limited to, vitamin A, vitamin E, vitamin C, retinal, tocopherol, L-ascorbyl palmitate, riboflavin, and any antioxidant suitable for oral administration, including carotenoids, including lutein, beta-carotene, zeaxanthin, and lycopene, and combinations thereof. Infant formulas may include oils, such as vegetable oils, including high oleic sunflower oil, coconut oil, canola oil, sunflower oil, or fish oil, and combinations thereof. Infant formulas may contain dairy products and milk-derived derivatives, including, for example, lactose, milk proteins, galactooligosaccharides, whey concentrate, fructooligosaccharides, and additional compounds concentrated from milk(s) secreted by mammals, including, but not limited to, humans, cows, etc.

[0106] Infant formulas may contain anti-caking agents (such as tricalcium phosphate, potassium chloride, sodium citrate, and potassium citrate), magnesium hydrogen phosphate, and coagulants (e.g., magnesium chloride, choline chloride, L-ascorbic acid, emulsifiers such as ferrous sulfate and zinc sulfate).

[0107] Although the powder may be reconstituted with water prior to use to a calorie density tailored to the nutritional needs of the end user, in most cases the powder is reconstituted with water to form a composition containing at least 19 kcal / fl oz (660 kcal / liter), more typically about 20 kcal / fl oz (675-680 kcal / liter) to about 25 kcal / fl oz (820 kcal / liter), and even more typically about 20 kcal / fl oz (675-680 kcal / liter) to about 24 kcal / fl oz (800-810 kcal / liter). Generally, formulas with 22-24 kcal / fl oz are more commonly used in preterm or low birth weight infants, while formulas with 20-21 kcal / fl oz (675-680-700 kcal / liter) are often used in full-term infants. In some embodiments, the reconstituted powder may have a caloric density of about 50-100 kcal / liter to about 660 kcal / liter, including about 150 kcal / liter to about 500 kcal / liter. In some specific embodiments, the emulsion may have a caloric density of 25, or 50, or 75, or 100 kcal / liter.

[0108] When the nutritional product is a powdered infant formula, the protein component is present in an amount of from about 5% to about 35%, including from about 8% to about 12%, and including from about 10% to about 12%, by weight of the infant formula; the fat component is present in an amount of from about 10% to about 35%, including from about 25% to about 30%, and including from about 26% to about 28%, by weight of the infant formula; and the carbohydrate component is present in an amount of from about 30% to about 85%, including from about 50% to about 55%, including from about 45% to about 60%, by weight of the infant formula.

[0109] The infant formulas contemplated herein may be formulated to include at least one of fat, protein, and carbohydrates, and preferably also include vitamins, minerals, and at least one compound of formula (I), (IA), (I-A1), (I-A2), (I-A3), (I-A4), (I-A5), and / or (I-A6), or combinations thereof.

[0110] There are numerous disease conditions in which plasmalogens have been demonstrated to play a protective role, including, but not limited to, metabolic disorders (obesity, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis), immune-related diseases (asthma, atopic dermatitis, type 1 diabetes, infectious diseases), cardiovascular diseases (atherosclerosis, cardiac remodeling, hypertension), neurological diseases (Alzheimer's disease, Parkinson's disease, autism, amyotrophic lateral sclerosis, frontotemporal dementia, multiple sclerosis, schizophrenia), cancer, myalgic encephalomyelitis / chronic fatigue syndrome, Barth syndrome, and peroxisomal disorders (Zellweger spectrum disorders, rhizomelic chondrodysplasia punctata). These are reviewed in several recent papers, including: Tremblay, et al., “Plasmalogens and platelet-activating factor roles in chronic inflammatory diseases,” BioFactors (2022) 1-14; Schooneveldt, et al., “Ether lipids in obesity: from cells to population studies,” Frontiers in Physiology (March 2022) 13:1-11; Boselli, Jr., et al., “Plasmalogen replacement therapy,” Membranes (2021) 11:838; and S. Paul, GI Lancaster, and P. Meikle, “Plasmalogens: a potential therapeutic target for neurodegenerative and cardiometabolic disease,” Progress Lipid Res. (2019) 74:186-195.

[0111] Supplement formulations are based on proportional levels of the corresponding PE(P) species in the circulation or tissues, and therefore, specific formulations can be used to target specific fluids (e.g., plasma), tissues such as the liver, heart, or adipose tissue, or specific cell types such as immune cells. For plasma targeting, one contemplated formulation contains ether lipids with a molar ratio of 18:1 alkenyl / alkyl R1 groups to 18:0 alkyl R1 groups to 16:0 alkyl R1 groups of about 1.0:1.7:1.4, with the molar percentage of 18:1 ether groups ranging from 18.6% to 27.9%, the molar percentage of 18:0 ether groups ranging from 32.6% to 45.8%, or the molar percentage of 16:0 ether groups ranging from 26.8% to 37.4%.

[0112] With respect to targeting immune cells, one embodiment of a contemplated formulation comprises an ether lipid having a molar ratio of 18:1 alkenyl / alkyl R1 groups to 18:0 alkyl R1 groups to 16:0 alkyl R1 groups of about 1.0:3.4:3.3.

[0113] Contemplated formulation embodiments include at least one compound of formula (I). In certain embodiments, the at least one compound of formula (I) is a compound of formula (IA). In certain embodiments, the at least one compound of formula (IA) is one or more of formula (I-A1), formula (I-A2), and / or formula (I-A3). In certain embodiments, the at least one compound of formula (I) is a mixture of two compounds of formula (I). In certain embodiments, the at least one compound of formula (I) is a mixture of two compounds of formula (IA). In certain embodiments, the at least one compound of formula (IA) is a mixture of two compounds selected from formula (I-A1), formula (I-A2), and formula (I-A3). In certain embodiments, the at least one compound of formula (IA) is a mixture of formula (I-A1), formula (I-A2), and formula (I-A3).

[0114] In certain embodiments, the mixture of Formula (I-A1), Formula (I-A2), and Formula (I-A3) comprises at least 50% of the ether lipids on a molar percentage basis. In certain embodiments, the mixture has a molar ratio of (I-A2) to (I-A1) of 1.2:1 to 2.5:1. In certain embodiments, the mixture has a molar ratio of (I-A2) to (I-A1) of 1.5:1 to 2.1:1. In certain embodiments, the mixture has a molar ratio of (I-A2) to (I-A1) of 1.7:1. In certain embodiments, the mixture has a molar ratio of (I-A2) to (I-A3) of 0.9:1 to 1.7:1. In certain embodiments, the mixture has a molar ratio of (I-A2) to (I-A3) of 1:1 to 1.5:1. In certain embodiments, the mixture has a molar ratio of (I-A2) to (I-A3) of 1.22:1. In certain embodiments, the mixture has a molar ratio of (I-A1) to (I-A3) of 0.5:1 to 1:1. In certain embodiments, the mixture has a molar ratio of (I-A1) to (I-A3) of 0.6:1 to 0.9:1. In certain embodiments, the mixture has a molar ratio of (I-A1) to (I-A3) of 0.72:1. In certain embodiments, the mixture has a molar percentage of (I-A1) of 18.6% to 27.9%, a molar percentage of (I-A2) of 32.6% to 45.8%, and a molar percentage of (I-A3) of 26.8% to 37.4%. In certain embodiments, the mixture has a molar percentage of (I-A1) of 23.3%, a molar percentage of (I-A2) of 39.2%, and a molar percentage of (I-A3) of 32.1%. In certain embodiments, the mixture has a molar ratio of (I-A1):(I-A2):(I-A3) of 1:1.7:1.4.

[0115] In certain embodiments, at least one compound of formula (I), (IA), (I-A1), (I-A2) and / or (I-A3) is converted in vivo to at least one plasmalogen.

[0116] Contemplated formulation embodiments include at least one compound of formula (I). In certain embodiments, the at least one compound of formula (I) is a compound of formula (IA). In certain embodiments, the at least one compound of formula (IA) is one or more of formula (I-A4), formula (I-A5), and / or formula (I-A6). In certain embodiments, the at least one compound of formula (I) is a mixture of two compounds of formula (I). In certain embodiments, the at least one compound of formula (I) is a mixture of two compounds of formula (IA). In certain embodiments, the at least one compound of formula (IA) is a mixture of two compounds selected from formula (I-A4), formula (I-A5), and formula (I-A6). In certain embodiments, the at least one compound of formula (IA) is a mixture of formula (I-A4), formula (I-A5), and formula (I-A6).

[0117] In certain embodiments, the mixture of Formula (I-A4), Formula (I-A5), and Formula (I-A6) comprises at least 50% of the ether lipids in the composition on a molar percentage basis. In certain embodiments, the mixture has a molar ratio of (I-A5) to (I-A4) of 1.2:1 to 2.5:1. In certain embodiments, the mixture has a molar ratio of (I-A5) to (I-A4) of 1.5:1 to 2.1:1. In certain embodiments, the mixture has a molar ratio of (I-A5) to (I-A4) of 1.7:1. In certain embodiments, the mixture has a molar ratio of (I-A5) to (I-A6) of 0.9:1 to 1.7:1. In certain embodiments, the mixture has a molar ratio of (I-A5) to (I-A6) of 1:1 to 1.5:1. In certain embodiments, the mixture has a molar ratio of (I-A5) to (I-A6) of 1.29:1. In certain embodiments, the mixture has a molar ratio of (I-A4) to (I-A6) of 0.5:1 to 1:1. In certain embodiments, the mixture has a molar ratio of (I-A4) to (I-A6) of 0.6:1 to 0.9:1. In certain embodiments, the mixture has a molar ratio of (I-A4) to (I-A6) of 0.76:1. In certain embodiments, the mixture has a molar percentage of (I-A4) of 18.6% to 27.9%, a molar percentage of (I-A5) of 32.6% to 45.8%, and a molar percentage of (I-A6) of 2.8% to 37.4%. In certain embodiments, the mixture has a molar percentage of (I-A4) of 24.8%, a molar percentage of (I-A5) of 42.4%, and a molar percentage of (I-A6) of 32.8%. In certain embodiments, the mixture has a molar ratio of (I-A4):(I-A5):(I-A6) of 1:1.7:1.4.

[0118] In certain embodiments, at least one compound of formula (I), (IA), (I-A4), (I-A5) and / or (I-A6) is converted in vivo to at least one plasmalogen.

[0119] In certain embodiments, the mixture of at least one compound of formula (I) or (IA) is a mixture of compounds of formula (I-A1), (I-A2), (I-A3), (I-A4), (I-A5) and / or (I-A6).

[0120] The disclosed ratios of the formulation are expected to produce optimized ratios of plasmalogens PE(P) and PC(P) in the serum and tissues of human subjects administered the formulation.

[0121] Other useful phosphatidyl classes, such as phosphatidylserine and phosphatidylinositol, can be prepared according to the principles herein.

[0122] In further embodiments, LPC(O), LPE(O), and LPA(O) can be used in various advantageous combinations that can be effective in increasing plasmalogen levels. The plasmalogens that are enhanced or modulated are generally represented by those listed above.

[0123] In certain embodiments, the formulation comprises one or more liquid or gel-based carriers, including but not limited to those selected from the group consisting of water and saline, urea, alcohols and their derivatives (e.g., methanol, ethanol, propanol, butanol), glycols (e.g., ethylene glycol, propylene glycol), and the like; natural or synthetic flavors and food-quality colorants; and thickeners, including but not limited to those selected from the group consisting of corn starch, guar gum, xanthan gum, and the like. In certain embodiments, the one or more liquid or gel-based carrier(s) may be added to the formulation at a weight / volume percentage of about 0.5% to about 95% weight / volume of the formulation. In certain embodiments, the natural or synthetic flavor(s) may be added to the formulation at a weight / volume percentage of about 3.0% to about 10.0% weight / volume of the formulation. In certain embodiments, the colorant(s) may be added to the formulation at a weight / volume percentage of about 1.0% to about 10.0% weight / volume of the formulation. In certain embodiments, a thickening agent may be added to the formulation at a weight / volume percentage of about 2% weight / volume of the formulation.

[0124] Delivery System The formulations disclosed herein can be delivered via dosage forms including, but not limited to, tablets, capsules, solutions, suspensions, powders, gums, and confectioneries. The formulations disclosed herein can be delivered via sublingual delivery systems including, but not limited to, sublingual and epilingual dissolving tablets, drops, beverages, and the like. Alternatively, or in addition, edible films, hydrophilic polymers, orally dissolving films, or orally dissolving strips can be used.

[0125] For oral administration, the formulations disclosed herein may be further combined with one or more solid inactive ingredients to prepare tablets, capsules, pills, powders, granules, or other suitable dosage forms. For example, the formulation components may be combined with at least one excipient, including but not limited to, fillers, binders, humectants, disintegrants, dissolution retarders, absorption accelerators, wetting agents, absorbents, and lubricants. Other useful excipients include, but are not limited to, magnesium stearate, calcium stearate, mannitol, xylitol, sweeteners, starch, carboxymethylcellulose, microcrystalline cellulose, silica, gelatin, silicon dioxide, etc. In some embodiments, formulations according to the present disclosure may include one or more of beeswax (such as Beeswax E901), carnauba wax (such as Carnauba Wax E903), shellac (such as Shellac E904), candelilla wax (such as Candelilla Wax E902), microcrystalline wax (such as Microcrystalline Wax E905), paraffin wax, and diacylglycerol.

[0126] The components of the formulations administered according to the methods of the present disclosure can be administered in a wide variety of oral dosage forms. It will be apparent to those skilled in the art that suitable dosage forms may, in certain embodiments, contain one or more compounds of the present disclosure and / or one or more pharmaceutically acceptable salts of the compounds of the present disclosure.

[0127] For preparing pharmaceutical formulations or compositions to be administered according to the methods of the present disclosure, pharmaceutically acceptable carriers can be either solid or liquid.Solid preparations include powders, tablets, pills, capsules, and cachets.A solid carrier can be one or more substances that can also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or encapsulating materials.

[0128] In powders, the carrier is a finely divided solid that is mixed with the finely divided active compound. In tablets, one or more compounds of the present disclosure and / or pharmaceutically acceptable salts thereof are mixed with one or more carriers having the necessary binding capacity in suitable proportions and then compacted to the desired shape and size.

[0129] In certain embodiments, powders and tablets administered according to the methods of the present disclosure may preferably contain, in total, about 1 to about 99 percent, e.g., 5 to about 70 percent, or 10 to about 70 percent, of one or more compounds of the present disclosure and / or pharmaceutically acceptable salts thereof. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low-melting waxes, cocoa butter, and the like. The term "preparation" is intended to include formulating one or more compounds of the present disclosure and / or pharmaceutically acceptable salts thereof with an encapsulating material as a carrier, with or without additional carrier, to provide a capsule in which one or more compounds of the present disclosure and / or pharmaceutically acceptable salts thereof are surrounded by, and thus associated with, the carrier. Cachets and lozenges are also included. Tablets, powders, capsules, pills, cachets, and lozenges are included. Tablets, powders, capsules, pills, cachets, lozenges, and fast-dissolve formulations can be used as solid forms suitable for oral administration.

[0130] Capsules can be prepared with an additional coating for sustained release. The thickness of the coating can be varied to provide delayed release of the capsule contents. Capsules can be prepared to be targeted release capsules. In some embodiments, capsules can be targeted to the stomach. In some embodiments, capsules targeted for delivery to the stomach are coated with a film coating. In some embodiments, capsules can be targeted to the small intestine. In some embodiments, capsules targeted for delivery to the small intestine are coated with an enteric coating.

[0131] As used herein, the term "fast dissolve" refers to a pharmaceutical formulation that dissolves upon contact with saliva, requiring little or no chewing.

[0132] Liquid preparations include, but are not limited to, solutions, suspensions, and emulsions, for example, water or water-propylene glycol solutions. The formulated preparations may take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending agents, stabilizing agents, solubilizing agents, and / or dispersing agents. Alternatively, one or more compounds of the present disclosure and / or pharmaceutically acceptable salts thereof may be in powder form, such as by aseptic isolation of a sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, for example, sterile, pyrogen-free water, before use.

[0133] Aqueous solutions suitable for oral use can be prepared by dissolving one or more compounds of the present disclosure and / or their pharmaceutically acceptable salts in water and adding suitable colorants, flavors, stabilizers and thickeners as needed. Aqueous suspensions suitable for oral use can be prepared by solubilizing and / or dispersing the finely divided active ingredient in water with viscous substances such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

[0134] Formulations suitable for topical, buccal, or sublingual administration in the mouth include, but are not limited to, lozenges comprising the active ingredient in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0135] In some embodiments, the formulation includes a solubilizing agent. As used herein, the term "solubilizing agent" refers to any agent that promotes the solubilization or dispersion of a composition when placed in a liquid. In some embodiments, a "solubilizing agent" may be a dispersing agent. A solubilizing agent may further improve the stability of the formulated composition. Suitable solubilizing agents include, but are not limited to, sodium carboxymethylcellulose, hypromellose, proline, xanthan gum, maltodextrin, alginate, wax, lipids, oils, alcohol, sugar, microcrystalline cellulose, starch, calcium phosphate, mannitol, sorbitol, erythritol, food-grade solvents, phospholipids such as lecithin (including, but not limited to, egg yolk L-α-lecithin, such as egg yolk L-α-lecithin available from Sigma-Aldrich, St. Louis, Missouri, USA), DMSO, ethanol, ethyl acetate, isopropanol, and the like. In some embodiments, a solubilizing agent is used to improve the separation of the compounds that make up the formulation and prevent precipitation or aggregation of the compounds in the formulation. In some embodiments, the formulation comprises a solubilizing agent and a suitable carrier. In some embodiments, such formulations are pharmaceutical-grade products. In some embodiments, such formulations are food-grade products.

[0136] Pharmaceutical formulations or preparations are preferably in unit dosage form.In this form, the preparation is subdivided into unit doses containing appropriate amounts of active ingredients.Unit dosage form can be a packaged preparation, and this package contains discrete amounts of preparations, such as packaged tablets, capsules, and powders in vials or ampoules.Also, unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or any of these in the appropriate number in packaged form.

[0137] Oral tablets, capsules, and lozenges, as well as oral liquids, are preferred formulations.

[0138] Further details on techniques for formulation and administration may be found in the latest edition of Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA).

[0139] Route of administration The compounds of the present disclosure and / or pharmaceutically acceptable salts thereof may be administered by any route, including, but not limited to, oral, sublingual, buccal, or as an oral spray.

[0140] The above-described methods may be further understood in connection with the following examples. The following non-limiting examples are provided to further illustrate the present disclosure. However, one of ordinary skill in the art will understand that it may be necessary to modify the procedures for any given embodiment of the present disclosure, e.g., to change the order or steps.

[0141] Numbered Embodiments 1. A composition comprising at least one compound of formula (I) or a pharmaceutically acceptable salt thereof [ka] 2.R 3 is selected from phosphates and substituted phosphates. 3.R 3The composition of embodiment 2, wherein is a substituted phosphate. 4. The composition of embodiment 3, wherein the substituted phosphate is substituted with an alkylamine or an inositol. 5. Alkylamines are [ka] and [ka] The composition of embodiment 4, selected from: 6.R 1 and R 2 are each independently hydrogen, optionally substituted C 1-30 alkyl groups, optionally substituted C 2-30 Alkenyl groups and optionally substituted C 1-30 The composition of any one of embodiments 1-5, wherein the acyl group is selected from: 7.R 1 and R 2 each independently represents an optionally substituted C 14-24 alkyl groups, optionally substituted C 14-24 Alkenyl groups and optionally substituted C 14-24 The composition of embodiment 6, wherein the acyl group is selected from: 8.R 2 is hydrogen, R 1 is an optionally substituted C 1-30 alkyl groups, optionally substituted C 2-30 alkenyl group, or optionally substituted C 1-30 The composition of embodiment 6, wherein the group is an acyl group. 9.R 1 is an optionally substituted C 14-24 alkyl groups, optionally substituted C 14-24 alkenyl group, or optionally substituted C 14-24 The composition of embodiment 8, wherein the group is an acyl group. 10.R 1 is an optionally substituted C 14-18 alkyl groups, optionally substituted C 14-18alkenyl group, or optionally substituted C 14-18 The composition of embodiment 9, wherein the group is an acyl group. 11.R 1 is the unsubstituted C 16 The composition of embodiment 10, wherein the group is an alkyl group. 12.R 1 is the unsubstituted C 18 The composition of embodiment 10, wherein the group is an alkyl group. 13.R 1 is the unsubstituted C 18 The composition of embodiment 10, wherein the alkenyl group. 14. The composition of embodiment 1, wherein at least one compound of formula (I) is a compound of formula (IA), or a pharmaceutically acceptable salt thereof. [ka] 15.Each R x is hydrogen or C 1-3 The composition of embodiment 14, wherein the alkyl is independently selected from: 16. The composition of embodiment 14 or 15, wherein n is 2 or 3. 17.R x The composition of embodiment 14, wherein is methyl and n is 3. 18.R A The composition of any one of embodiments 14-17, wherein is an optionally substituted hydrocarbon chain containing 14 to 24 carbon atoms. 19.R A The composition of embodiment 18, wherein is an optionally substituted hydrocarbon chain containing 16 to 18 carbon atoms. 20.R A is an optionally substituted C 16-18 alkyl groups, optionally substituted C 16-18 alkenyl group, or optionally substituted C 16-18 The composition of embodiment 19, wherein the group is an acyl group. 21.R A is an optionally substituted C 16-18 The composition of embodiment 20, wherein the group is an alkyl group. 22.R A is an optionally substituted C16-18 The composition of embodiment 20, wherein the alkenyl group. 23.R A is the unsubstituted C 16 The composition of embodiment 19, wherein the group is an alkyl group. 24.R A is the unsubstituted C 18 The composition of embodiment 19, wherein the group is an alkyl group. 25.R A is the unsubstituted C 18 The composition of embodiment 19, wherein the alkenyl group. 26. The composition of any of embodiments 1-25, wherein the composition comprises a mixture of at least two compounds of formula (I) and / or (IA). 27. The composition of any of embodiments 1-26, wherein the composition comprises a mixture of three compounds of formula (I) and / or (IA). 28. The composition of embodiment 14, wherein the compound of formula (IA) is of structure (I-A1), (I-A2), or (I-A3). [ka] 29. The composition of embodiment 28, wherein the composition comprises a mixture of at least two compounds selected from the group consisting of (I-A1), (I-A2), and (I-A3). 30. The composition of embodiment 28, wherein the composition comprises a mixture of (I-A1), (I-A2), and (I-A3). 31. The composition of embodiment 28, wherein the composition comprises a mixture of (I-A1), (I-A2), and (I-A3), wherein the mixture of (I-A1), (I-A2), and (I-A3) accounts for at least 50% of the ether lipids in the composition on a molar percentage basis. 32. The composition of embodiment 28, wherein the mixture has a molar ratio of (I-A2) to (I-A1) of 1.2:1 to 2.5:1. 33. The composition of embodiment 32, wherein the mixture has a molar ratio of (I-A2) to (I-A1) of 1.5:1 to 2.1:1. 34. The composition of embodiment 33, wherein the mixture has a molar ratio of (I-A2) to (I-A1) of 1.7:1. 35. The composition of embodiment 28, wherein the mixture has a molar ratio of (I-A2) to (I-A3) of 0.9:1 to 1.7:1. 36. The composition of embodiment 35, wherein the mixture has a molar ratio of (I-A2) to (I-A3) of 1:1 to 1.5:1. 37. The composition of embodiment 36, wherein the mixture has a molar ratio of (I-A2) to (I-A3) of 1.22:1. 38. The composition of embodiment 28, wherein the mixture has a molar ratio of (I-A1) to (I-A3) of 0.5:1 to 1:1. 39. The composition of embodiment 38, wherein the mixture has a molar ratio of (I-A1) to (I-A3) of 0.6:1 to 0.9:1. 40. The composition of embodiment 39, wherein the mixture has a molar ratio of (I-A1) to (I-A3) of 0.72:1. 41. The composition of embodiment 30, wherein the mixture has a molar percentage of (I-A1) between 18.6% and 27.9%, a molar percentage of (I-A2) between 32.6% and 45.8%, and a molar percentage of (I-A3) between 26.8% and 37.4%. 42. The composition of embodiment 30, wherein the mixture has a molar percentage of (I-A1) of 23.3%, a molar percentage of (I-A2) of 39.2%, and a molar percentage of (I-A3) of 32.1%. 43. The composition of embodiment 30, wherein the mixture has a molar ratio of (I-A1):(I-A2):(I-A3) of 1:1.7:1.4. 44. The composition of embodiment 14, wherein the compound of formula (IA) is of structure (I-A4), (I-A5), or (I-A6). [ka] 45. The composition of embodiment 44, wherein the composition comprises a mixture of at least two compounds selected from the group consisting of (I-A4), (I-A5), and (I-A6). 46. ​​The composition of embodiment 44, wherein the composition comprises a mixture of (I-A4), (I-A5), and (I-A6). 47. The composition of embodiment 44, wherein the composition comprises a mixture of (I-A4), (I-A5), and (I-A6), wherein said mixture of (I-A4), (I-A5), and (I-A6) accounts for at least 50% of the ether lipids in the composition, on a molar percentage basis. 48. The composition of embodiment 44, wherein the mixture has a molar ratio of (I-A5) to (I-A4) of 1.2:1 to 2.5:1. 49. The composition of embodiment 48, wherein the mixture has a molar ratio of (I-A5) to (I-A4) of 1.5:1 to 2.1:1. 50. The composition of embodiment 49, wherein the mixture has a molar ratio of (I-A5) to (I-A4) of 1.7:1. 51. The composition of embodiment 44, wherein the mixture has a molar ratio of (I-A5) to (I-A6) of 0.9:1 to 1.7:1. 52. The composition of embodiment 51, wherein the mixture has a molar ratio of (I-A5) to (I-A6) of 1:1 to 1.5:1. 53. The composition of embodiment 52, wherein the mixture has a molar ratio of (I-A5) to (I-A6) of 1.29:1. 54. The composition of embodiment 44, wherein the mixture has a molar ratio of (I-A4) to (I-A6) of 0.5:1 to 1:1. 55. The composition of embodiment 54, wherein the mixture has a molar ratio of (I-A4) to (I-A6) of 0.6:1 to 0.9:1. 56. The composition of embodiment 55, wherein the mixture has a molar ratio of (I-A4) to (I-A6) of 0.76:1. 57. The composition of embodiment 46, wherein the mixture has a molar percentage of (I-A4) between 18.6% and 27.9%, a molar percentage of (I-A5) between 32.6% and 45.8%, and a molar percentage of (I-A6) between 26.8% and 37.4%. 58. The composition of embodiment 57, wherein the mixture has a molar percentage of (I-A4) of 24.8%, a molar percentage of (I-A5) of 42.4%, and a molar percentage of (I-A6) of 32.8%. 59. The composition of embodiment 46, wherein the mixture has a molar ratio of (I-A4):(I-A5):(I-A6) of 1:1.7:1.3. 60. A formulation, such as a pharmaceutical formulation, comprising a composition as defined in any one of embodiments 1-59 and at least one excipient. 61. The formulation of embodiment 60, wherein the formulation is formulated for oral administration. 62. The formulation of embodiment 61, wherein the oral dosage form is a tablet, capsule, solution, mouthwash, suspension, powder, gum, confectionery, lozenge, sublingual delivery system, or fast-dissolving formulation. 63. The formulation of any one of embodiments 60-62, wherein the pharmaceutical formulation is a food product. 64. The formulation of embodiment 63, wherein the formulation is a dietary supplement. 65. The formulation of embodiment 63, wherein the food is a medical food. 66. The formulation of embodiment 63, wherein the food is an infant formula. 67. A formulation according to any one of embodiments 60 to 66 for maintaining or regulating plasmalogen levels in a human subject in need thereof. 68. A formulation according to any one of embodiments 60 to 66 for maintaining or regulating plasmalogen levels in an animal subject in need thereof. 69. The formulation of embodiment 68, wherein the animal is selected from domestic animals, working animals and farm animals. 70. A formulation according to any one of embodiments 67 to 69, wherein said maintaining or regulating requires maintaining or regulating the level of plasmalogens at levels and / or ratios associated with a non-disease state. 71. A composition as defined in any one of embodiments 1-59, or a formulation as defined in any one of embodiments 60-70, for use in increasing the level of a plasmalogen compound in the blood or tissue of a subject. 72. The composition for use according to embodiment 71, wherein the subject is a human. 73. The composition for use according to embodiment 71, wherein the subject is an animal. 74. The composition for use according to embodiment 73, wherein the animal subject is selected from domestic animals, working animals and farm animals. 75. A composition as defined in any one of embodiments 1-59, or a formulation as defined in any one of embodiments 60-70, for use in therapy. 76. A composition as defined in any one of embodiments 1-59, or a formulation as defined in any one of embodiments 60-70, for use in treating a disease or disorder associated with a deficiency of plasmalogens. 77. The composition or formulation for use according to embodiment 76, wherein the disease or disorder is a neurological disease. 78. The composition or formulation of embodiment 77, wherein the neurological disease is selected from Alzheimer's disease, Parkinson's disease, autism, amyotrophic lateral sclerosis, frontotemporal dementia, multiple sclerosis, and schizophrenia. 79. A composition or formulation for use according to embodiment 76, wherein the disease or disorder is a metabolic disorder. 80. The composition or formulation of embodiment 79, wherein the metabolic disorder is selected from the group consisting of obesity, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, immune-related diseases, cardiovascular diseases, neurological diseases, cancer, myalgic encephalomyelitis / chronic fatigue syndrome, Barth syndrome, and peroxisomal disorders. 81. The composition or formulation of embodiment 80, wherein the immune-related disease is selected from the group consisting of asthma, atopic dermatitis, type 1 diabetes, and infectious diseases. 82. The composition or formulation of embodiment 80, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cardiac remodeling, and hypertension. 83. The composition or formulation of embodiment 80, wherein the peroxisomal disorder is Zellweger syndrome spectrum disorder or rhizomelic chondrodysplasia punctata. 84. A composition or formulation for use according to any one of embodiments 71-83, wherein the compound of formula (I) or (IA) is administered in a dose of 0.1 to 4000 mg per day. 85. The composition or formulation for use according to embodiment 84, wherein the compound of formula (I) or (IA) is administered in a dose of 0.1 to 2000 mg per day. 86. The composition or formulation for use according to embodiment 85, wherein the compound of formula (I) or (IA) is administered in a dose of 25 to 1600 mg per day. 87. The composition or formulation for use according to embodiment 86, wherein the compound of formula (I) or (IA) is administered in a dose of 400 mg, 800 mg, or 1600 mg per day. 88. The composition or formulation for use according to embodiment 86, wherein the compound of formula (I) or (IA) is administered in a dose of 200 mg per day. 89. The composition or formulation for use according to embodiment 86, wherein the compound of formula (I) or (IA) is administered in a dose of 25 to 100 mg per day. 90. A method for increasing the level of a plasmalogen compound in the blood or tissue of a subject in need thereof, comprising administering to the subject an effective amount of at least one compound of formula (I) or a pharmaceutically acceptable salt thereof, and / or at least one compound of formula (IA) or a pharmaceutically acceptable salt thereof, as defined in embodiments 1 and 14, respectively. 91. The method of embodiment 90, wherein the subject is a human. 92. The method of embodiment 90, wherein the subject is an animal. 93. The method of embodiment 92, wherein the animal subject is selected from domestic animals, working animals, and farm animals. 94. The method of embodiment 90, wherein the bioavailability of the plasmalogen compound in the blood or tissues of the subject is improved after administration of at least one compound of formula (I) or a pharmaceutically acceptable salt thereof, and / or at least one compound of formula (IA) or a pharmaceutically acceptable salt thereof, compared to administration of AKG (alkylglycerol). 95. A method for treating a disease or disorder associated with a deficiency of plasmalogens, comprising administering to a subject in need thereof an effective amount of at least one compound of formula (I) or a pharmaceutically acceptable salt thereof, and / or at least one compound of formula (IA) or a pharmaceutically acceptable salt thereof, as defined in embodiments 1 and 14, respectively. 96. The method of embodiment 95, wherein the subject is a human. 97. The method of embodiment 95, wherein the subject is an animal. 98. The method of embodiment 97, wherein the animal subject is selected from domestic animals, working animals, and farm animals. 99. The method of embodiment 95, wherein the disease or disorder is a neurological disease. 100. The method of embodiment 99, wherein said neurological disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, autism, amyotrophic lateral sclerosis, frontotemporal dementia, multiple sclerosis, and schizophrenia. 101. The method of embodiment 95, wherein the disease or disorder is a metabolic disorder. 102. The method of embodiment 101, wherein the metabolic disorder is selected from the group consisting of obesity, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, immune-related diseases, cardiovascular diseases, neurological diseases, cancer, myalgic encephalomyelitis / chronic fatigue syndrome, Barth syndrome, and peroxisomal disorders. 103. The method of embodiment 102, wherein the immune-related disease is selected from the group consisting of asthma, atopic dermatitis, type 1 diabetes, and infectious diseases. 104. The method of embodiment 102, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cardiac remodeling, and hypertension. 105. The method of embodiment 102, wherein the peroxisomal disorder is Zellweger syndrome spectrum disorder or rhizomelic chondrodysplasia punctata. 106. The method of any one of embodiments 90-105, wherein the compound of formula (I) or (IA) is administered at a dose of 0.1 to 4000 mg per day. 107. The method of embodiment 106, wherein the compound of formula (I) or (IA) is administered at a dose of 0.1 to 2000 mg per day. 108. The method of embodiment 107, wherein the compound of formula (I) or (IA) is administered at a dose of 25 to 1600 mg per day. 109. The method of embodiment 108, wherein the compound of formula (I) or (IA) is administered at a dose of 400 mg, 800 mg, or 1600 mg per day. 110. The method of embodiment 108, wherein the compound of formula (I) or (IA) is administered at a dose of 200 mg per day. 111. The method of embodiment 108, wherein the compound of formula (I) or (IA) is administered at a dose of 25 to 100 mg per day. 112. The method of embodiment 111, wherein the compound of formula (I) or (IA) is administered at a dose of 25 mg, 50 mg, or 100 mg per day. 113. Use of a composition described in any one of embodiments 1 to 59 or a formulation described in any one of embodiments 60 to 70 in the manufacture of a medicament for increasing the level of a plasmalogen compound in the blood or tissue of a subject in need thereof. 114. Use of a composition according to any one of embodiments 1 to 59 or a formulation according to any one of embodiments 60 to 70 in the manufacture of a medicament for treating a disease or disorder associated with a deficiency of plasmalogens. 115. The use according to embodiment 114, wherein the disease or disorder is a neurological disease. 116. The use according to embodiment 115, wherein the neurological disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, autism, amyotrophic lateral sclerosis, frontotemporal dementia, multiple sclerosis, and schizophrenia. 117. The use according to embodiment 116, wherein the disease or disorder is a metabolic disorder. 118. The use according to embodiment 117, wherein the metabolic disorder is selected from the group consisting of obesity, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, immune-related diseases, cardiovascular diseases, neurological diseases, cancer, myalgic encephalomyelitis / chronic fatigue syndrome, Barth syndrome, and peroxisomal disorders. 119. The use according to embodiment 118, wherein the immune-related disease is selected from the group consisting of asthma, atopic dermatitis, type 1 diabetes, and infectious diseases. 120. The use according to embodiment 118, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cardiac remodeling, and hypertension. 121. The use according to embodiment 118, wherein the peroxisomal disorder is Zellweger syndrome spectrum disorder or rhizomelic chondrodysplasia punctata. 122. A kit for use in the method of any one of embodiments 90 to 112, comprising at least one compound of formula (I) or formula (IA), or a pharmaceutically acceptable salt thereof, as defined in any one of embodiments 1 and 14, respectively. 123. The composition of embodiment 28, wherein at least one compound is selected from compounds of formula (I-A1), (I-A2) or (I-A3). 124. The composition of embodiment 29, wherein the mixture of at least two compounds is a 50:50 mixture. 125. A mixture of (I-A1), (I-A2) and (I-A3) LPC(O-16:0)26.8%+LPC(O-18:0)46.1%+LPC(O-18:1)27.1%, LPC(O-16:0)46%+LPC(O-18:0)21%+LPC(O-18:1)33%, LPC(O-16:0)42%+LPC(O-18:0)51%+LPC(O-18:1)7%, LPC(O-16:0)62%+LPC(O-18:0)23.9%+LPC(O-18:1)14.1%, LPC(O-16:0)19.8%+LPC(O-18:0)66%+LPC(O-18:1)14.2%, LPC(O-16:0) 32.5% + LPC(O-18:0) 39.6% + LPC(O-18:1) 27.9%, or 31. The composition of embodiment 30, wherein the LPC is selected from: 34.1% LPC(O-16:0) + 41.5% LPC(O-18:0) + 24.4% LPC(O-18:1). 126. The composition of embodiment 44, wherein at least one compound is selected from compounds of formula (I-A4), (I-A5) or (I-A6). 127. The composition of embodiment 45, wherein the mixture of at least two compounds is a 50:50 mixture. 128. A mixture of (I-A4), (I-A5) and (I-A6) LPE(O-16:0)26.8%+LPE(O-18:0)46.1%+LPE(O-18:1)27.1%, LPE(O-16:0)46%+LPE(O-18:0)21%+LPE(O-18:1)33%, LPE(O-16:0)42%+LPE(O-18:0)51%+LPE(O-18:1)7%, LPE(O-16:0)62%+LPE(O-18:0)23.9%+LPE(O-18:1)14.1%, LPE(O-16:0)19.8%+LPE(O-18:0)66%+LPE(O-18:1)14.2%, LPE(O-16:0)32.5%+LPE(O-18:0)39.6%+LPE(O-18:1)27.9%, or 47. The composition of embodiment 46, wherein the LPE is selected from: 34.1% LPE(O-16:0) + 41.5% LPE(O-18:0) + 24.4% LPE(O-18:1). 129. The composition of any one of embodiments 1-59 or 123-128, wherein the composition maintains or modulates the plasmalogen compound ratio observed in healthy human subjects. 130. The composition of any one of embodiments 1-59 or 123-128, wherein the composition results in a reduction in inflammation or an improvement or reduction in symptoms associated with an inflammatory disease. 131. The composition of embodiment 130, wherein the reduction in inflammation is associated with a reduction in the levels of inflammatory cytokines. 132. The composition of any one of embodiments 1-59 or 123-128, wherein the composition maintains or regulates the levels of inflammatory cytokines at levels observed in healthy human subjects. 133. The composition of embodiment 132, wherein the inflammatory cytokines that are maintained or regulated are selected from IL-6, NFE2L2, TLR4. 134. A formulation comprising the composition according to any one of embodiments 1 to 59 or 123 to 128 and at least one excipient and / or solubilizer. 135. The formulation of embodiment 134, wherein the formulation is formulated for oral administration. 136. The formulation of embodiment 135, wherein the oral dosage form is a drink or food. 137. The formulation of embodiment 135, wherein the oral dosage form is a food product. 138. The formulation of embodiment 135, wherein the oral dosage form is an infant formula. 139. The formulation of embodiment 134, wherein the formulation comprises a solubilizer. 140. The formulation of embodiment 139, wherein the solubilizer is selected from the group consisting of sodium carboxymethylcellulose, hypromellose, proline, xanthan gum, maltodextrin, alginate, wax, lipid, oil, alcohol, sugar, microcrystalline cellulose, starch, calcium phosphate, mannitol, sorbitol, erythritol, food grade solvents, phospholipids, DMSO, ethanol, ethyl acetate, and isopropanol. 141. The formulation of embodiment 139, wherein the solubilizer is lecithin. 142. The formulation of embodiment 141, wherein the lecithin is purified egg yolk L-α-lecithin. 143. The formulation of embodiment 139, wherein the formulation has improved stability compared to a formulation comprising a composition defined in any one of embodiments 1 to 59 or 123 to 128 that does not contain a solubilizer. 144. The formulation of embodiment 139, wherein the formulation is less susceptible to degradation during freezing and thawing compared to a formulation comprising a composition as defined in any one of embodiments 1 to 59 or 123 to 128 that does not contain a solubilizer. 145. The composition defined in any one of embodiments 28 or 44, wherein the composition comprises one compound having a purity of at least 99.9%. 146. A formulation comprising the composition defined in embodiment 145 and at least one excipient. 147. The formulation defined in any one of embodiments 134-138 or embodiment 146, wherein the formulation comprises at least one of a solubilizer, an emulsifier, a stabilizer, a dispersant, an antifoaming agent, or a diluent. 148. The formulation defined in any one of embodiments 134-138 or embodiments 146-147, wherein the formulation further comprises at least one solubilizer. 149. The formulation of embodiment 148, wherein the solubilizer is selected from the group consisting of sodium carboxymethylcellulose, hypromellose, proline, xanthan gum, maltodextrin, alginate, wax, lipid, oil, alcohol, sugar, microcrystalline cellulose, starch, calcium phosphate, mannitol, sorbitol, erythritol, food grade solvents, phospholipids, DMSO, ethanol, ethyl acetate, and isopropanol. 150. The formulation of embodiment 149, wherein the solubilizer is selected from the group consisting of DMSO, ethanol, ethyl acetate, and isopropanol. 151. A formulation as defined in any one of embodiments 134-144 or embodiments 146-150, wherein the formulation further comprises at least one antioxidant compound. 152. A formulation as defined in any one of embodiments 134-144 or embodiments 146-151, wherein the formulation further comprises at least one antifoaming agent. 153. The formulation defined in any one of embodiments 134-144 or embodiments 146-152, wherein the formulation is formulated for oral administration. 154. The formulation of embodiment 153, wherein the oral dosage form is a tablet, capsule, solution, mouthwash, suspension, powder, gum, confectionery, lozenge, sublingual delivery system, or fast-dissolving formulation. 155. The formulation of embodiment 154, wherein the oral dosage form is a tablet. 156. The composition of embodiment 30, wherein the mixture of (I-A1), (I-A2) and (I-A3) comprises 90% w / v of (I-A1). 157. The composition of embodiment 30, wherein the mixture of (I-A1), (I-A2) and (I-A3) comprises 90% w / v of (I-A2). 158. The composition of embodiment 30, wherein the mixture of (I-A1), (I-A2) and (I-A3) comprises 90% w / v of (I-A3). 159. The composition of embodiment 46, wherein the mixture of (I-A4), (I-A5) and (I-A6) comprises 90% w / v of (I-A4). 160. The composition of embodiment 46, wherein the mixture of (I-A4), (I-A5) and (I-A6) comprises 90% w / v of (I-A5). 161. The composition of embodiment 46, wherein the mixture of (I-A4), (I-A5) and (I-A6) comprises 90% w / v of (I-A6).

[0142] The following is a non-limiting example: Two studies were compared to determine the effect of precursor compounds and various formulations of the precursor compounds on plasmalogen levels in humans.

[0143] Example 1 The first study showed the effect of krill oil and fish oil supplementation on plasmalogen levels in female subjects.

[0144] Details of lipidomics analysis are fully described in K. Huynh, et al., "High-Throughput Plasma Lipidomics: Detailed Mapping of the Associations with Cardiometabolic Risk Factors," Cell Chem. Biol. (2019) 26(1):71-84, and H. H. Sung, et al., "Differential plasma postprandial lipidomic responses to krill oil and fish oil supplements in women: A randomized crossover study," Nutrition (2019) 65:191-201. Briefly, plasma samples were extracted in CHCl3:MeOH (2:1) with an internal standard mixture containing non-physiological or stable isotope-labeled lipid standards, as previously described. Lipidomic analysis was performed by UHPLC ESI-MS / MS using an Agilent 1290 HPLC coupled to an Agilent 6490 triple quadrupole mass spectrometer. Chromatographic data were analyzed using Mass Hunter Quant, and relative lipid abundance was calculated by relating the area under each lipid species' chromatogram to the corresponding internal standard. Correction factors were applied to adjust for different response factors, if known. Chromatographically separated species were labeled accordingly (e.g., PC(16:0 / 22:6) and PC(18:2 / 20:4)), while mixed isomer species were given standard phospholipid designations (e.g., PC(40:8) was a mixture of PC(20:4 / 20:4) and PC(18:2 / 22:6)). When structural details were sufficient, lipids were manually annotated as containing long-chain omega-3 components (i.e., 20:5 EPA, 22:5 DPA, and 22:6 DHA).

[0145] This study was a randomized crossover study with 30-day supplementation with krill oil (KO) and fish oil (FO), separated by a minimum 4-week washout period. Participants were instructed to maintain their usual diet and were required not to consume any foods or supplements containing omega-3 PUFAs more than once a week during the study period. For the intervention, participants consumed either seven 1-gram KO capsules (Antarctic Krill Oil, Swiss Wellness Pty Ltd., Victoria, Australia) containing 1.27g of LC omega-3 PUFAs (0.76g EPA, 0.42g DHA, 0.09g DPA) or five 1-gram FO capsules (Natural FO, Swiss Wellness Pty Ltd., Victoria, Australia) containing 1.44g of LC omega-3 PUFAs (0.79g EPA, 0.47g DHA, 0.18g DPA) daily for 30 days. Participants were required to visit the clinic three times for blood and data collection on days 0 (baseline), 15, and 30 for each supplementation period. Before each clinic visit, participants were advised to consume a low-fat dinner, avoid alcohol and strenuous physical activity, and fast from 10:00 PM. On each study day, participants arrived at the clinic between 7:00 AM and 9:00 AM, and a standardized procedure was performed in which a fasting blood sample (10 mL) was collected via venipuncture by a certified phlebotomist. After blood sample collection, all participants completed a 24-hour dietary recall and an electronic PUFA FFQ. The study protocol was approved by the Ethics Committee of Victoria University Human Research (HRE15-031). Informed consent was obtained from all participants prior to the study (ACTRN 12615000472572).

[0146] Example 2 The second study showed the effect of shark liver oil (SLO) supplementation on plasmalogen levels in male subjects.

[0147] This was a double-blind, placebo-controlled, crossover study. Participants (n = 10) were overweight or obese (28–40 kg / m) without evidence of cardiovascular disease or diabetes. 2 The participants were adult males (25-60 years old) with a BMI in the range of 0.01 to 0.01. Written informed consent was obtained from all study participants prior to the start of the study. This study was conducted in accordance with the ethical principles set out in the Declaration of Helsinki and received approval from the Alfred Hospital Ethics Committee (approval number 436 / 15). Participants were randomly assigned to a placebo or treatment group and received 4 g of Alkyrol® (purified SLO, Eurohealth, Ireland) or placebo (methylcellulose) per day for 3 weeks, followed by a 3-week washout period, and then crossed over to a 3-week alternating placebo / Alkyrol® treatment. Fasting blood samples were collected at the beginning and end of each intervention. Tables 3 and 4 show the composition of alkyl diacylglycerols [TG(O)] and alkyl glycerols (AKG), respectively, in Alkyrol® shark liver oil. [Table 3] [Table 4]

[0148] The data sets from the studies in Examples 1 and 2 were compared to extrapolate the effects of these oil supplements on plasma ether lipid levels. Figures 1 and 2 show that both KO and SLO supplementation substantially increased the levels of different ether lipid classes in plasma, while FO supplementation had little effect on these lipid classes. The increase in plasmalogens (PE(P) and PC(P)) was significantly higher with KO supplementation (30 days) than with SLO supplementation (21 days).

[0149] The effects of supplementation on PE(P) composition were also extrapolated. Figures 3 and 4 show that neither KO nor FO supplementation significantly affected the alkenyl chain composition of PE plasmalogens, whereas Figure 5 shows that SLO supplementation significantly altered this composition (increasing the proportion of 18:1 alkenyl chains containing PE plasmalogens and decreasing the proportion of 16:0 and 18:0 alkenyl chains containing PE plasmalogens). All of these supplementations had substantial effects on the acyl chain composition of PE plasmalogens. Figures 6 and 7 show that both KO and FO supplementation decreased the proportion of 18:1, 18:2, and 20:4 alkenyl chains containing PE plasmalogens and increased the proportion of 20:5 and 22:6 alkenyl chains containing PE plasmalogens, whereas Figure 8 shows that SLO supplementation decreased the proportion of 20:4 alkenyl chains containing PE plasmalogens and increased the proportion of 18:1 and 22:6 alkenyl chains containing PE plasmalogens.

[0150] Without wishing to be bound by theory, these differences may arise from the type and content of ether lipids found in KO and SLO. In KO, the ether lipids are primarily phosphatidylcholine ether lipids (PC(O) or LPC(O)), whereas in SLO, the ether lipids are monoalkyl-diacylglycerols or TG(O). KO was found to contain approximately 25 mg of LPC(O) / g oil, whereas SLO contained approximately 200 mg of alkylglycerol / g oil. Furthermore, the alkyl chain composition of KO and SLO differs. In SLO, the most abundant alkyl species is O-18:1 (Paul, et al., Journal of Lipid Research (2021) 62:10009), whereas in KO, the most abundant alkyl species is O-16:0 (Table 4). [Table 5]

[0151] The relative bioavailability of KO and SLO for ether lipid modulation is further compared in Table 6. Taking into account the changes in ether lipid levels, KO LPC(O) appears to have significantly higher bioavailability than SLO alkylglycerols. [Table 6]

[0152] We observed that KO resulted in a 12-fold higher rate of change in PE(P) levels per micromolar dose compared to SLO, and this effect was nearly retained (8-fold higher) when adjusted over the treatment period. This represents a significant improvement in the bioavailability of LPC(O) / PC(O) in krill oil compared to alkyl diacylglycerols in SLO.

[0153] Example 3 Animal experiments were performed by Product Safety Labs (Dayton, NJ, USA). A total of 32 animals (all female) were used. Animals were selected based on adequate weight gain and the absence of clinical signs of disease or injury. The selected rats were randomized and assigned to the following test groups according to weight stratification, such that the mean weight of the groups did not differ by more than 20% within sex across groups (Table 7). Animals had free access to food and water throughout the experiment. Each animal was dosed by oral intubation using a stainless steel ball-tip gavage needle attached to an appropriate syringe. Dose administration was performed once on day 1. [Table 7]

[0154] Plasmalogen precursor compounds and preparations Stable isotope versions of four different lipid species were administered at two doses (C, D) (Table 8). All of these compounds were synthesized by Anthem Biosciences (Bengaluru, India). [Table 8]

[0155] To facilitate administration of the precursor compounds, purified egg yolk L-α-lecithin (Sigma-Aldrich, St. Louis, MO, USA) was used as a vehicle (14 mg for Dose Mixture C and 47 mg for Dose Mixture D). Stock solutions of each precursor (100 mg / ml) were prepared in chloroform:methanol (1:1). A stock solution of lecithin (100 mg / ml, Sigma-Aldrich, St. Louis, MO, USA) was also prepared in chloroform:methanol (1:1). To prepare the precursor compound mixture, appropriate amounts of precursor compound and lecithin solution were mixed, dried under a stream of nitrogen gas at 40°C, and then reconstituted in deionized water by vigorous vortexing, followed by sonication for 1 h in an ultrasonic cleaner water bath (Soniclean, Adelaide, South Australia, Australia) and further sonication for 2 × 30 s at an amplitude of 25 using a Misonix S-4000 sonicator (Thermo Fisher Scientific, Melbourne, Victoria, Australia).

[0156] Taking a blood sample Blood samples were collected from all animals at eight time points (pre-dose, 1, 2, 4, 8, 12, 24, and 48 hours post-dose). Approximately 200 μL of blood was collected sublingually under isoflurane anesthesia into a blood collection tube containing K2EDTA and kept on ice until centrifugation. After centrifugation, plasma was transferred to clean tubes and frozen at approximately -80°C until analysis.

[0157] Terminal sacrifice and tissue collection Fecal samples from 24- and 48-hour periods were collected at terminal sacrifice, and all surviving animals were euthanized using CO2 asphyxiation. After 48 hours, the brain, liver, spleen, kidney, heart, gastrocnemius muscle, and abdominal adipose tissue from all study animals were carefully dissected, placed in new weighing boats, wet weighed, and then snap-frozen within 5 minutes and stored at -80°C. No other observations were made on any animals at necropsy.

[0158] Lipid extraction from plasma samples Lipids were extracted from 10 μl plasma samples using butanol:methanol (1:1) as previously described (Alshehry ZH.Et al 2015). Lipid extraction was performed in a single batch, with quality control samples (pooled plasma QC, NIST QC, and blank) included every 20 samples.

[0159] Tissue processing and lipid extraction from tissue samples Approximately 40–60 mg of tissue was homogenized in 400–600 μL of ice-cold phosphate-buffered saline using a TissueLyser II (Qiagen, USA) for 60 seconds, followed by sonication for 10–15 seconds at an amplitude of 25 using a Misonix S-4000 sonicator (Thermo Fisher Scientific, Melbourne, Victoria, Australia). The protein content of the homogenate was quantified using a Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific, Rockford, IL, USA). Homogenates were then prepared to a stock protein concentration of 5 mg / mL, and a 10 μL aliquot from the stock solution containing 50 μg of protein was subsequently used for lipid extraction. For adipose tissue, a 10 μL aliquot from the stock homogenate was used for lipid extraction. Lipids were extracted according to a previously described single-phase extraction procedure with chloroform:methanol (2:1) (Miekle PJ et al. 2011). Lipid extraction was performed as a single batch for each tissue type, with the exception of liver and brain samples, which were extracted together, with quality control samples (pooled plasma QC, pooled tissue QC, NIST QC, and blank) included approximately every 10 samples.

[0160] Liquid chromatography-mass spectrometry Lipidomic analysis was performed on an Agilent 1290U HPLC system (ZORBAX eclipse plus C18 column: 2.1 x 100 mm, 1.8 mm, Agilent) and an Agilent 6495 triple quadrupole mass spectrometer, set at 45 °C. Samples were placed in an autosampler tray (20 °C) and 1 μL was injected. Lipids were separated at a flow rate of 0.4 mL / min using a stepwise linear gradient of solvent A (50% water / 30% acetonitrile / 20% isopropanol) and solvent B (1% water / 9% acetonitrile / 90% isopropanol), both containing 10 mM ammonium formate. The solvent gradient started at 85% A, decreased to 50% over 2.5 min, 43% over 0.1 min, 30% over 6.4 min, 7% over 0.1 min, 4% over 1.9 min, then decreased to 0% over 0.1 min, held for 0.9 min, then increased again to 85% over 0.2 min and held for 3.8 min for equilibration. The following mass spectrometer conditions were used: gas temperature 150°C, gas flow rate 17 L / min, nebulizer 20 psi, sheath gas temperature 200°C, capillary voltage 3500 V, and sheath gas flow rate 10 L / min.

[0161] A modified version of a previously reported MRM method (K. Huynh, et al., "High-Throughput Plasma Lipidomics: Detailed Mapping of the Associations with Cardiometabolic Risk Factors," Cell Chem. Biol. (2019) 26(1):71-84) was used to detect deuterium incorporation into lipid species of the ether phospholipid and ether glycerolipid classes: alkyl diacylglycerol [TG(O)], lysoalkylphosphatidylcholine [LPC(O)], alkylphosphatidylcholine [PC(O)], alkylphosphatidylethanolamine [PE(O)], alkenylphosphatidylethanolamine [PE(P)] or PE-plasmalogen, and alkenylphosphatidylcholine [PC(P)] or PC-plasmalogen). Both deuterated and endogenous unlabeled versions of lipids were measured. Lipids with deuterated isotopes are designated [+1] and [+2], while endogenous lipids are referred to as [+0]. D2-labeled precursors generate both [+1] and [+2] lipid species due to the position of the deuterium, making them susceptible to removal by the Δ1 desaturase (PEDS1) enzyme during plasmalogen formation. The precursor masses of D2-labeled lipid species were offset by 2 Da, since one deuterium corresponds to 1 Da. Product ions were offset based on the fragmentation patterns of the lipid classes.

[0162] Using Agilent MassHunter quantitative analysis software, we integrated the peaks and quantified the area under the curve for each lipid species. To calculate the relative concentration of deuterium-labeled lipids in the samples, we first subtracted the signal of the lipid species in the blank from the sample. This eliminated any background signal captured by the mass spectrometer. Next, we calculated the isotope ratio in the time 0 sample (tissue from the control male sample), which corresponds to the area of ​​deuterium-labeled lipids relative to the area of ​​endogenous lipids. This allowed us to calculate the natural isotope ratio of each lipid to be subtracted. Next, we multiplied the isotope ratio by the background-subtracted area of ​​each lipid to obtain the endogenous signal of each lipid. This value was then subtracted from the area of ​​all deuterium-labeled lipids. We then divided this value by the area of ​​the internal standard for the corresponding lipid class and multiplied it by the amount of internal standard added to the sample. This gave us the relative concentration of deuterated lipids in the sample.

[0163] Effect of supplementation of labeled precursors on plasma ether lipids We observed distinct patterns in the concentrations of various tracer lipids over 48 h after oral administration of the precursor compounds. Specifically, TG(O) concentrations reached their peak within 1–2 h of administration and then rapidly declined in all treatment groups (Figures 29A–H). In the case of PC(O) (Figures 31A–H), it took approximately 8–12 h to reach maximum concentrations for all groups except LPC(O)-Dose Mixture D (Figure 30H) (24 h). Interestingly, it took much longer (12–24 h) for LPC(O) concentrations to reach peak levels in all treatment groups, followed by a gradual decline (Figures 30A–H). For PE(O) (Figures 32A–H), the AKDAG-DHA treatment produced a much steeper trajectory (a rapid rise within 8 h followed by a rapid decline) compared to the other treatments (Figures 30C and 30G). The trajectories of PE(P) were very gradual (Figures 33A-H). In particular, LPC(O) showed a much gradual trajectory compared to the other groups, with its concentration peaking at the 24-h mark and then gradually declining (Figures 33D and 33H), but still exhibiting significant amounts of tracer PE(P) at 48 h. Tracer PC(P) concentrations (Figures 34A-H) peaked at 24 h in all groups except LPC(O) after fluctuations at earlier time points (Figures 34D and 34H) and remained stable until 48 h. In the LPC(O) group, tracer PC(P) concentrations gradually increased from 12 h to 48 h. These observations highlight that the timelines of the lipid concentration peak and subsequent decline differ across treatments and highlight their differential dynamics of absorption and metabolism.

[0164] Next, we compared the maximum plasma tracer concentrations (Cmax) of the major ether lipid classes between the different experimental groups (Figures 35A-35F). We observed that the highest Cmax was consistently associated with the LPC(O) group across multiple lipid classes, except for TG(O) (Figure 35A). As expected, we observed a dose-dependent increase in plasma LPC(O) levels after treatment with LPC(O) (Figure 35B). Furthermore, a similar dose-dependent pattern was observed for PC(O) (Figure 35C) and PE(O) (Figure 35D) within the LPC(O) group. Furthermore, the LPC(O) group showed a significantly higher Cmax of PE(P) compared to the other groups (Figure 35E). The Dose Mixture D LPC(O) group also showed an elevated Cmax of PC(P) (Figure 35F).

[0165] Although LPC(O) was much more efficient than the other compounds in generating PE(P), the actual conversion was low. Treatment with LPC(O) dose mixture C converted to PC(O) at Cmax, representing 1.4% present in plasma (Figure 35C), but converted to PE(P) at Cmax, representing only 0.12% present in plasma (Figure 35F).

[0166] Next, we calculated the total exposure of the labeled precursor compound as the area under the curve (AUC) using the tracer lipid concentrations at different time points (Figures 36A-36F). We did not observe significant differences in AUC between the AKG, AKDAG with oleic acid [AKDAG(OA)], and AKDAG with DHA [AKDAG(DHA)] groups in PE(P) production (Figure 36E), particularly in the case of PE(P) for dose mixture C of each compound. However, LPC(O) administration (Figure 36B) resulted in an approximately 5-fold increase in PE(P) compared with AKG and AKDAG, based on a comparison between dose mixture C groups. Interestingly, LPC(O) showed rapid metabolism to PC(O), resulting in a significant 16-fold increase in PC(O) levels (Figure 36C), accompanied by a relatively modest 3-fold increase in PE(O) (Figure 36D). This observation is intriguing and indicates that PC(O) is a metabolic sink for LPC(O). Unlike LPC(O), AKG and AKDAG(OA) treatments showed rapid metabolism to TG(O) with significantly less conversion to PC(O), PE(O), and PE(P). On the other hand, AKDAG(DHA), particularly in dose mixture D, showed significant conversion to PE(P), although much less than LPC(O). Notably, PC(P) production (Figure 36F) remained limited across all treatments within the 2-day time frame.

[0167] Effect of supplementation of labeled precursors on tissue ether lipids In addition to analyzing plasma lipids, we examined tissue ether lipids to assess the degree of incorporation of labeled precursors into endogenous ether lipids. Our observations indicate that administration of LPC(O) effectively converted labeled precursors to newly synthesized LPC(O), PC(O), and PE(O) in the liver, spleen, brain, and kidney (Figures 37A-F). We did not observe significant incorporation of labeled precursors into endogenous ether lipids in visceral adipose tissue, skeletal muscle, or heart (Figures 41A-F).

[0168] Example 4 Plasmalogen precursor therapy Cells and mice were treated with plasmalogen precursors and their bioavailability and conversion to endogenous plasmalogens were compared. Precursors labeled with different numbers of deuterium atoms at various positions were used due to their availability. The number of deuterium-labeled sites is indicated using the letter "D" followed by a number. For example, "D2" refers to a precursor labeled with two deuterium atoms (see Scheme III). For example, cells were treated with AKG (O-16:0, D2) (Anthem Biosciences, Bengaluru, India), LPC (O-16:0, D4) (Cayman Chemical, Ann Arbor, USA), and LPE (O-16:0, D5) (Avanti Polar Lipids, Birmingham, USA) (Table 9). These are deuterium-labeled plasmalogen precursors containing 16:0 alkenyl chains (Scheme III). For comparison, mice were administered a mixture of AKG (O-16:0, O-18:0, O-18:1, D2) (Anthem Biosciences, Bengaluru, India), a mixture of LPC (O-16:0, O-18:0, O-18:1, D2) (Anthem Biosciences, Bengaluru, India), and LPE (O-16:0, D5) (Avanti Polar Lipids, Birmingham, USA) (Table 9). The mixture of AKG and LPC(O) was deuterium-labeled and contained 16:0, 18:0, and 18:1 alkenyl chains (Scheme III). LPE(O) contained a mixture of deuterium-labeled 16:0 alkenyl chains and unlabeled 16:0, 18:0, and 18:1 alkenyl chains (Scheme III). [Table 9] [ka] Scheme III. Deuterium-labeled plasmalogen precursors used in this study: alkylglycerol (AKG), lysophosphatidylcholine (LPC(O)), and lysophosphatidylethanolamine (LPE(O)).

[0169] cell culture Cell lines were selected based on tissues commonly affected by diseases associated with low plasmalogen levels. The two cell lines were the immortalized human hepatocellular carcinoma HepG2 and the immortalized mouse fibroblast 3T3. Cells were cultured at 37°C and 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (Gibco, USA) supplemented with 1% sodium pyruvate (Gibco, USA) and either 10% fetal bovine serum (Gibco, USA) for HepG2 or 10% newborn calf serum (Gibco, New Zealand) for 3T3. HepG2 cells were passaged weekly, with the medium replenished every 2–3 days. After 30 passages, these cells were discarded. 3T3 cells were passaged every 2–3 days and discarded after 39 passages. Both cell lines were cultured for at least two passages before dissemination occurred. HepG2 cells were seeded in 12-well plates at 300,000 cells / ml of medium to achieve 80% confluency, whereas 3T3 cells reached 100% confluency before the experiment.

[0170] treatment To compare the uptake and incorporation of plasmalogen precursors into plasmalogens, cells were treated with deuterium-labeled AKG, LPC(O), and LPE(O) at a concentration of 20 μM (Table 9). Because AKG is dissolved in ethanol, the control treatment was medium containing 0.05% ethanol. Before treatment, images were taken using an Olympus CKX41 inverted microscope, and cells were analyzed for Ca. 2+ and Mg 2+The cells were then washed with phosphate-buffered saline (PBS) without AKG. Figures 9 and 13 show pre- and post-treatment images of HEPG2 and 3T3 cells. The cells were then treated with AKG, LPC (0), LPE (0), or control and incubated for 24 hours. After incubation, images were taken, and only adherent cells were collected. For HepG2, the cells were incubated with trypsin-ethylenediaminetetraacetic acid (EDTA) (0.05%) and phenol red (Gibco, USA) for 5 minutes at 37°C and 5% CO2. They were then centrifuged at 13,000 g for 15 minutes. The supernatant was then removed, and the remaining pellet was dried using a high-speed vacuum (Thermo Scientific). For 3T3, the cells were washed with PBS and a cell scraper was used. The cell suspension was dried using a high-speed vacuum pump. After the drying process, the samples were stored at -80°C until lipid extraction was performed.

[0171] Animal experiments To compare the bioavailability and incorporation of plasmalogen precursors into plasmalogens, mice were administered deuterium-labeled AKG mixture, LPC(O) mixture, and LPE(O) (Table 9). Lecithin was used as a vehicle to facilitate oral delivery and as a control treatment. Animal care and experiments were approved by the Alfred Health and Education Precinct Animal Ethics Committee (P8490). The study included 64 approximately 8-week-old C57BL / 6 mice, consisting of 32 males and 32 females. Figure 9 shows the administration schedule for 64 8-week-old C57BL / 6 mice treated with a single dose of deuterium-labeled precursors and controls via oral gavage. Precursors included alkylglycerol (AKG), lysoalkylphosphatidylcholine (LPC(O)), and lysoalkylphosphatidylethanolamine (LPE(O)). Blood samples were collected by tail bleeding and via cardiac puncture at 0, 1, 4, 24, and 48 hours. Mice were housed at the Precinct Animal Center of the Baker Heart and Diabetes Institute with a 12-hour light / dark cycle and unlimited access to standard chow (SF00-105) and water. There were eight treatment groups, each consisting of four females and four males, which received a single dose of either Dose Mixture A or Dose Mixture B of lecithin (vehicle control), AKG, LPC(O), and LPE(O) (Table 10). Prior to treatment, mice were randomly assigned to treatment groups based on body weight and fasted for 4 hours. Each mouse received 200 μl of each treatment by oral gavage. Blood was collected by tail bleeding before treatment and at 1, 4, 24, and 48 hours after treatment. At the end of the experiment, mice were anesthetized with an intraperitoneal injection of sodium pentobarbital, and blood was collected by cardiac puncture. Mice were then euthanized by cervical dislocation, and tissues were harvested and flash-frozen. Blood samples were collected in EDTA tubes and stored on ice. Plasma was separated from the blood by centrifugation at 1,710 g and 20°C for 15 minutes. Plasma supernatants were stored at -80°C until lipid extraction. [Table 10]

[0172] lipid extraction Lipid extraction was performed to harvest lipids from cell and plasma samples for lipidomic analysis. As part of the extraction process, pooled blanks and plasma quality control samples (PQCs) from plasma of healthy individuals were extracted every 10–20 samples. Additionally, mouse plasma extraction included a NIST quality control (NIST QC) and mouse plasma quality control (mouse PQC) using plasma from two females and two males from each group. These quality control samples helped identify any variations during the extraction process. Depending on the sample type, lipids were extracted using chloroform:methanol (2:1) or butanol:methanol (BUME, 1:1) based on previously reported methods (Alshehry ZHet al., Metabolites, 2015 Jun 17;5(2):389-403; Meikle PJ, Wong G, Tsorotes D, Barlow CK, Weir JM, Christopher MJ, et al., "Plasma lipidomic analysis of stable and unstable coronary artery disease," Arteriosclerosis, Thrombosis, and Vascular Biology. 2011,31(11):2723-32). To calculate relative lipid concentrations, internal standards containing known concentrations of over 20 non-physiological lipids were mixed with either chloroform:methanol or butanol:methanol before use in the extraction.

[0173] Extraction of HepG2 cells involved combining 10 μl of sample, blank, and PQC with 200 μl of chloroform:methanol mixed with an internal standard. The samples were then rotated on a rotary mixer for 10 minutes, sonicated in a water bath for 30 minutes, and allowed to stand at room temperature for 20 minutes. The samples were then centrifuged at 13,000 rpm for 10 minutes and dried using a high-speed vacuum pump. The extracted lipids were then reconstituted in 50 μl of water-saturated butanol, sonicated for 10 minutes, and 50 μl of methanol was added. Finally, the samples were centrifuged at 4,000 rpm for 5 minutes, and the supernatant was transferred to a glass vial with a Teflon glass insert (Agilent) and stored at -80°C for lipidomic analysis.

[0174] Extraction of 3T3 cells involved combining 10 μl of sample, blank, and PQC with 100 μl of BUME mixed with the internal standard. The samples were then vortexed for 10 seconds, sonicated in a water bath for 60 minutes, and centrifuged at 13,000 rpm for 10 minutes. The supernatant was then transferred to a glass vial with a Teflon glass insert for lipidomic analysis.

[0175] BUME was also used for mouse plasma extraction, but only 5 μl of sample mixed with 5 μl of water was used. Samples included plasma supernatant, blank, PQC, NIST QC, and mouse PQC. Because only 5 μl of sample was used, half the concentration of the internal standard was mixed with BUME compared to the HepG2 and 3T3 cell extractions.

[0176] To extract lipids from brain samples, approximately 50 mg of brain tissue was homogenized in 500 μL of ice-cold phosphate-buffered saline using a TissueLyser II (Qiagen, USA) for 60 seconds and sonicated for 10–15 seconds at an amplitude of 25 using a Misonix S-4000 sonicator (Thermo Fisher Scientific, Melbourne, Victoria, Australia). The protein content of the homogenate was quantified using a Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific, Rockford, IL, USA). The homogenate was then prepared to a stock protein concentration of 5 mg / mL, and a 10 μL aliquot from the stock solution containing 50 μg of protein was subsequently used for lipid extraction. Lipids were extracted according to a previously described single-phase extraction procedure using chloroform:methanol (2:1) (Meikle PJ, Wong G, Tsorotes D, Barlow CK, Weir JM, Christopher MJ, et al., "Plasma lipidomic analysis of stable and unstable coronary artery disease," Arteriosclerosis, Thrombosis, and Vascular Biology. 2011, 31(11):2723-32). Lipid extraction was performed as a single batch, with quality control samples (pooled plasma QC, pooled tissue QC, NIST QC, and blank) included approximately every 10 samples.

[0177] Lipidomics analysis by UHPLC / MS / MS Targeted lipidomic analysis was performed on lipid extracts of HepG2 cells, 3T3 cells, and plasma samples using high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC / MS / MS). Prior to analysis, the lipid extracts were randomized, thawed at room temperature for 1 hour, and sonicated in a water bath for 15 minutes. Lipids were quantified using an Agilent 1290 Series UHPLC system coupled with an Agilent 6495C triple quadrupole mass spectrometer. First, samples were placed in an autosampler tray at 20°C, and either 1 μl of cell sample or 2 μl of plasma sample was injected. Lipids were separated with a stepwise linear gradient of solvent A and solvent B under the liquid chromatography conditions outlined in Tables 11 and 12. Lipids were then identified based on their retention times and the masses of precursor and product ions under the mass spectrometry conditions outlined in Table 13. [Table 11] [Table 12] [Table 13]

[0178] A modified version of a previously reported MRM method was used to detect deuterium incorporation into lipid species of the glycerophospholipid and glycerolipid classes (Huynh, K. et al., 2019). Both deuterated and endogenous unlabeled versions of lipids were measured. Lipids bearing deuterated isotopes are designated [+1], [+2], [+4], and [+5], while endogenous lipids are referred to as [+0]. D4-labeled precursors generate [+4] lipid species, while D5-labeled precursors result in [+5] lipid species. However, D2-labeled precursors generate both [+1] and [+2] lipid species due to the position of the deuterium, making them susceptible to removal by the PEDS1 enzyme during plasmalogen generation. The precursor masses of D4-labeled lipid species were offset by 4 Da, since one deuterium atom corresponds to 1 Da. Product ions were offset based on the fragmentation pattern of the lipid class.

[0179] The peaks were integrated and the area under the curve for each lipid was quantified using Agilent MassHunter Quantitative Analysis v10.0 software. To calculate the relative concentration of deuterium-labeled lipids in cell samples, we first subtracted the background signal area from the blank sample. This eliminated any extraneous background signal identified by the mass spectrometer. Next, we calculated the isotope ratio in the control sample, which corresponds to the area of ​​deuterium-labeled lipids relative to the area of ​​endogenous lipids. This allowed us to calculate the natural isotope of each lipid to be removed. Next, we multiplied the isotope ratio of each sample by the subtracted background area to obtain the endogenous signal for each lipid. This value was then subtracted from the area of ​​all deuterium-labeled lipids. We then divided this value by the area of ​​the internal standard for the corresponding lipid class and multiplied it by the amount of internal standard added. This provided the relative concentration of deuterium-labeled lipids in pmol / sample. Any variation in lipid concentration between samples, such as differences in cell number, was removed by normalizing to PC34:1, one of the most abundant phospholipids. The relative concentration of PC34:1 was calculated by dividing the area of ​​PC34:1 by the area of ​​its internal standard. This was then multiplied by 100 to obtain the concentration in μmol / sample. The relative concentration of the deuterated lipid was then divided by the relative concentration of PC34:1 to obtain the concentration in pmol / μmol of PC34:1. The relative concentration of each lipid class was also calculated as the sum of the individual species within the class. Only neutral loss species were considered when calculating the total TG(O).

[0180] Relative lipid concentrations in mice were calculated by multiplying the deuterated lipid concentration (μmol / sample) by 200 to obtain concentrations in pmol / ml of plasma before normalizing to PC34:1. Additionally, for calculation of LPE(O), concentrations were divided by 0.65 because both labeled and unlabeled precursors were present.

[0181] Effect of plasmalogen precursors on the plasmalogen biosynthetic pathway in HepG2 cells To compare the uptake and incorporation of plasmalogen precursors into plasmalogens, HepG2 cells were treated with 20 μM deuterium-labeled plasmalogen precursors for 24 hours and analyzed for lipid concentrations using LC-MS / MS. Images taken before and after treatment revealed that LPC(O) and LPE(O) resulted in floating cells and smaller adherent cells, whereas AKG did not affect cell morphology compared to the control (Figure 9).

[0182] Levels of labeled PE(P) of 16:0, 18:0, and 18:1 species were compared between treatment groups. Concentrations were found to be significantly higher in the plasmalogen precursor group compared to controls (p ≤ 0.01, Figure 10A). This increase in labeled PE(P) was most pronounced in LPE(O), followed by LPC(O), and then AKG (Figure 10A). To better understand how efficiently precursors are metabolized to plasmalogens, the ratio of labeled PE(P) concentration to total labeled concentration was calculated. Here, total labeling refers to the sum of labeled AKG, alkyl-acylglycerol (DG(O)), LPC(O), lysophosphatidylcholine plasmalogen (LPC(P)), LPE(O), lysophosphatidylethanolamine plasmalogen (LPE(P)), alkylphosphatidylcholine (PC(O)), phosphatidylcholine plasmalogen (PC(P)), alkylphosphatidylethanolamine (PE(O)), PE(P), and monoalkyl-diacylglycerol (TG(O)). This approach accounts for the amount of precursor taken up by cells and is therefore a better representation of precursor efficiency. Some precursors may have greater uptake, resulting in higher production of PE(P), but this does not necessarily mean they are more efficient at producing PE(P) when requiring more precursor. Comparison of plasmalogen precursor groups showed that the ratio of LPE(O) was significantly greater than both AKG and LPC(O) (p≦0.0001, Figure 10B). Furthermore, the ratio of AKG was significantly increased compared to LPC(O) (p≦0.05, Figure 10B). However, the difference between AKG and LPC(O) was smaller than the difference between LPE(O) and the other precursors.

[0183] Bar graphs were generated to visualize the distribution of label among lipid classes in the plasmalogen biosynthetic pathway (Figure 11). After AKG treatment, the majority of label was incorporated into AKG and DG(O), accounting for 40% and 23% of the total label, respectively (Figures 11A and 12A). In comparison, incorporation into TG(O), LPC(O), LPE(O), and PC(P) was minimal, with moderate incorporation into PC(O), PE(O), and PE(P), with PE(P) comprising 11% of the total label (Figures 11A and 12A). After LPC(O) treatment, incorporation into LPC(O) was high and even higher into PC(O), accounting for 53% of the total label (Figures 11B and 12B). There was moderate incorporation into AKG, DG(O), and PE(P), with PE(P) comprising 7% of the total label (Figures 11B and 12B). The remaining lipid classes showed little or no incorporation (Figure 11B). After LPE(O) treatment, there was moderate incorporation into AKG, LPE(O), and PC(O), and higher incorporation into PE(O) and PE(P) (Figure 11C). The majority of the label was incorporated into PE(P), accounting for 35% of the total label (Figure 12C). Otherwise, incorporation into DG(O) was low, and incorporation into TG(O), LPC(O), and PC(P) was minimal (Figure 11C).

[0184] To verify the results with HepG2 cells, 3T3 cells were treated with 20 μM deuterium-labeled plasmalogen precursor for 24 hours and analyzed for lipid concentrations using LC-MS / MS. Unlike HepG2 cells, plasmalogen precursor treatment in 3T3 cells did not result in changes in cell morphology compared to controls (Figure 13). By comparing labeled PE(P) concentrations between treatment groups, we found that concentrations significantly increased after plasmalogen precursor treatment compared to controls (p≦0.01, Figures 14A and 14B). This increase was most pronounced in AKG, followed by LPE(O), and then LPC(O) (Figure 14A). However, when comparing the ratio of labeled PE(P) concentration to total labeled concentration, we observed that LPE(O) was significantly higher than both AKG and LPC(O), which is consistent with the results from HepG2 cells (Figure 14C). Although LPC(O) had a higher ratio than AKG, this difference was much smaller than the difference between LPE(O) and AKG (FIG. 14C).

[0185] Bar graphs were also generated to assess label distribution within the plasmalogen biosynthetic pathway (Figure 15). After AKG treatment, a high proportion of label was incorporated into AKG, accounting for 55% of the total label (Figures 15A and 16A). This finding was consistent with the results observed in HepG2 cells. Furthermore, incorporation into PC(O) and PE(O) was moderate, while incorporation into the remaining lipid classes was minimal, with only 11% of the total label incorporated into PE(P) (Figures 15A and 16A). Comparing the effects of AKG treatment across cell lines, a lower proportion of label was found to be incorporated into PE(P) and DG(O) in 3T3 cells compared to HepG2 cells (Figures 11A and 15A). After LPC(O) treatment, there was high incorporation into LPC(O) and moderate incorporation into PC(O), which was opposite to the results observed in HepG2 cells (Figures 11B and 15B). There was little or no incorporation into the remaining lipid classes, with only 4% of the total label incorporated into PE(P) (Figures 15B and 16B). After LPE(O) treatment, there was moderate incorporation into LPE(O) and higher incorporation into PC(O), PE(O), and PE(P) (Figure 15C). In HepG2 cells, the majority of the label was incorporated into PE(P), whereas in 3T3 cells, the majority was incorporated into PE(O), accounting for 34% of the total label (Figures 12C and 16C). Incorporation into PE(P) was also high, accounting for 29% of the total label (Figure 16C). The remaining lipid classes showed little to moderate incorporation (Figure 15C).

[0186] To compare the bioavailability of plasmalogen precursors and their incorporation into plasmalogens, mice were treated with a single dose of deuterium-labeled plasmalogen precursors (Figure 17). Plasma lipid concentrations from multiple time points were analyzed using LC-MS / MS. Unlike cell lines, LPE(O) treatment only used labeled 16:0, compared to AKG and LPC(O), which contained 16:0, 18:0, and 18:1 lipids. Therefore, lipidomic analysis in mice considered only 16:0 lipid species. Although lipid concentrations were measured for all treated mice, the data reported here primarily focus on Dose Mixture A-treated mice, as Dose Mixture B-treated mice showed similar trends.

[0187] To estimate the rate at which AKG is taken up into the circulation, the levels of labeled AKG were compared between treatment groups. In females, high concentrations of labeled AKG were present for all precursors 1 hour after treatment (Figures 18A, 19A, and 20A). However, in males, AKG treatment resulted in a more pronounced increase in labeled AKG 1 hour after treatment compared with LPC(O) and LPE(O) (Figures 18B, 19B, and 20B). Furthermore, AKG treatment in males resulted in approximately twice the amount of labeled AKG compared with females (Figures 18A and 18B).

[0188] Because plasmalogen precursors are first converted to these lipids before generating plasmalogens, we measured the levels of labeled PC(O) and PE(O). For all plasmalogen precursors, the highest concentrations of labeled PC(O) occurred 4 hours after treatment (Figures 19A and 19B). Furthermore, treatment with LPC(O) resulted in the greatest increase in labeled PC(O), followed by LPE(O) and AKG (Figures 19A and 19B). Similar to PC(O), labeled PE(O) concentrations were highest for all plasmalogen precursors after 4 hours of treatment (Figures 20A and 20B). Treatment with LPE(O) resulted in the greatest increase in labeled PE(O), followed by LPC(O) and AKG (Figures 20A and 20B).

[0189] Examination of labeled PE(P) concentrations revealed that LPE(O) treatment resulted in the greatest increase, followed by LPC(O) and then AKG (Figures 21A, 21B, 22A, 22B). This was observed 24 hours after treatment and was consistent in females and males receiving either the therapeutic dose of Dose Mixture A (Figures 21A, 21B) or Dose Mixture B (Figures 22A, 22B).

[0190] To evaluate the bioavailability of plasmalogen precursors, the C of PE(P) max In both males and females treated with Dose Mixture A, LPC(O) and LPE(O) treatments showed significantly higher C than control and AKG. max Furthermore, LPE(O) treatment showed significantly higher C compared to LPC(O) (p≦0.01, Figures 23A, 23B, 23C, and 23D). max Males treated with LPE(O) had 1.45-fold higher C than females (p≦0.001, Figures 23A and 23C). max A similar trend was evident in mice treated with Dose Mixture B (Figures 23E, 23F, and 23G, 23H). In both males and females, LPE(O) treatment resulted in significantly increased C compared to control and AKG. max Furthermore, LPE(O) treatment resulted in significantly greater C than LPC(O) in males (p≦0.05, Figures 23E and 23G). max Treatment with LPC(O) resulted in a greater C than AKG, although this difference was only observed as a trend in females (p≦0.0001, Figures 23E and 23G). max Although this was only seen as a trend in mice treated with Dose Mixture B compared to Dose Mixture A (Figures 23A-23H), comparing mice treated with different doses, Dose Mixture B of LPE(O) produced 1.77-fold higher C than Dose Mixture A in females. max In comparison, Dose Mixture B of LPE(O) was found to result in a 1.55-fold greater C than Dose Mixture A in males (Figures 23A and 23E). max(Figures 23C and 23G).

[0191] The bioavailability of plasmalogen precursors was further evaluated by comparing the AUC of PE(P) between groups. max Similar results were obtained in both males and females treated with dose mixture A of LPC(O) or LPE(O) that showed significantly higher AUC than the control or AKG (p≦0.001, Figures 24A, 24B, 24C, and 24D). Furthermore, LPE(O) treatment resulted in a significantly greater AUC than LPC(O) (p≦0.0001, Figures 24A and 24C). It was further observed that LPE(O) treatment in males resulted in a 1.3-fold higher AUC than in females (Figures 24A and 24C). Similarly, these trends were reflected in mice treated with dose mixture B. LPE(O) treatment resulted in a higher AUC than LPC(O), AKG, or the control in both males and females (p≦0.05, Figures 24E, 24F, 24G, and 24H). LPC(O) treatment resulted in a greater AUC than AKG (Figures 24E and 24G). Comparing mice treated with different doses, it was observed that LPE(O) dose mixture B produced a 1.5-fold higher AUC than dose mixture A in both males and females (Figures 24A-24H).

[0192] AUC bar graphs for females and males treated with Dose Mixture A were generated to visualize the distribution of label across the plasmalogen biosynthetic pathway (Figures 25 and 26). In females, AKG treatment resulted in high incorporation into DG(O) and TG(O), and moderate incorporation into PC(O) and PE(O) (Figure 25A). There was also minimal or no incorporation into AKG, LPC(O), LPE(O), PE(P), and PC(P) (Figure 25A). A similar trend was evident after AKG treatment in males (Figure 26A). In comparison, LPC(O) treatment in females resulted in moderate incorporation into DG(O), TG(O), LPC(O), and PE(P), and high incorporation into PC(O) and PE(O) (Figure 25B). AKG, LPE(O), and PC(P) showed little or no incorporation (Figure 25B). LPC(O) treatment resulted in a greater increase in TG(O) in males, while other lipid classes showed similar trends to females (Figures 25B and 26B). After LPE(O) treatment in both females and males, incorporation into PE(O) was high, incorporation into PC(O) and PE(P) was moderate, and incorporation into the remaining lipid classes was minimal or nonexistent (Figures 25C and 26C).

[0193] tissue analysis The conversion of labeled precursor compounds to brain PE plasmalogens was examined, and it was observed that LPE(O) supplementation resulted in a small but significant increase in labeled PE plasmalogens in the brain (Figure 27), whereas the increase in labeled PE(P) with AKG and LPC(O) supplementation was quite variable (Figure 28).

[0194] statistical analysis Lipidomic analysis was performed on triplicates of HepG2 and 3T3 cells and four replicates of plasma samples. However, samples collected from males treated with lecithin dose mix A had five replicates at 0 h post-treatment. This was because one mouse died before treatment but was still included in the analysis. Figures were created using Microsoft Excel and presented as mean ± standard deviation. Statistical analysis was performed using GraphPad Prism 9.5.1. One-way analysis of variance (ANOVA) was performed to compare treatment groups, followed by Tukey's post-hoc test to determine which groups were significantly different. Significance was indicated by *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, and ****p ≤ 0.0001. p < 0.05 was considered statistically significant.

[0195] Example 5 This example relates to the modulation of plasmalogen levels and composition by supplementation of LPC(O) and LPE(O) in RAW264.7 cells with different SN-1 alkyl compositions.

[0196] Cell culture method This study used the murine macrophage cell line RAW264.7. Cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium containing 1% sodium pyruvate, L-glutamine (Gibco, USA), and 10% fetal bovine serum (Gibco, USA) at 37°C with 5% CO2. Cells were passaged weekly, with the medium replenished every 2–3 days until passage 30, at which point the cells were discarded. Cells were seeded in 12-well plates at a concentration of 300,000 cells / ml of medium and grown until they reached 80% confluency.

[0197] Replenishing cells with plasmalogen precursors To evaluate the incorporation of plasmalogen precursors into cellular plasmalogens, cells were supplemented with compounds with different SN1 compositions, i.e., LPC(O) or LPE(O) at various ratios of O-16:0, O-18:0, and O-18:1, at a concentration of 20 μM (Table 14). The plasmalogen precursors, LPC(O) and LPE(O), were dissolved in chloroform:methanol (2:1), evaporated at 40°C under a stream of nitrogen gas, and then reconstituted in RPMI medium containing 2% bovine serum albumin (Sigma-Aldrich, USA). The control treatment for cells consisted of RPMI medium containing 2% bovine serum albumin. Once prepared, the precursor-containing medium was vortexed and sonicated for 15 minutes in an ultrasonic water bath (Soniclean, Adelaide, South Australia, Australia). [Table 14]

[0198] lipid extraction Lipids were extracted from cells using the butanol-methanol method according to established protocols. Each lyophilized cell sample received 10 μl of MilliQ water and 100 μl of a butanol-methanol mixture (1:1) containing a specific set of internal standards. After this, each sample was vortexed for a few seconds and placed in an ultrasonic bath for 1 hour. The samples were then centrifuged at 13,000 g for 15 minutes, and the resulting supernatant was carefully transferred to a mass spectrometry vial with an insert. These vials were then frozen at -80°C until ready for mass spectrometry analysis.

[0199] lipid analysis A targeted lipidomics approach using LC-MS / MS according to a previously published protocol (Hynh, et al., "High-Throughput Plasma Lipidomics: Detailed Mapping of the Associations with Cardiometabolic Risk Factors," Cell Chem. Biol., 2019 Jan. 17;26(1):71-84.e4, doi:10.1016 / j.chembiol.2018.10.008) was utilized for the samples in this study. The method uses liquid chromatography (Agilent 1290 Infinity) coupled with tandem mass spectrometry (Agilent 6495C) operated in dynamic multiple reaction monitoring (MRM) mode. Multiple lipid species belonging to different lipid classes were measured, including phosphatidylcholine (PC), alkylphosphatidylcholine [PC(O)], alkenylphosphatidylcholine [PC(P)], phosphatidylethanolamine (PE), alkylphosphatidylethanolamine [PE(O)], alkenylphosphatidylethanolamine [PE(P)] or PE-plasmalogen, lysophosphatidylcholine (LPC), lysoalkylphosphatidylcholine [LPC(O)], lysoalkenylphosphatidylcholine [LPC(P)], lysophosphatidylethanolamine (LPE), lysoalkylphosphatidylethanolamine [LPE(O)], lysoalkenylphosphatidylethanolamine [LPE(P)], triacylglycerol (TG), and alkyldiacylglycerol [TG(O)]. The lipid species measured in this study are listed in Supplementary Table 1 included at the end of this example.

[0200] The chromatographic gradient utilized in this analysis included a series of steps: starting at 15% B at a flow rate of 0.4 mL / min, increasing to 50% B over 2.5 min, followed by 57% over 0.1 min, 70% over 6.4 min, 93% over 0.1 min, and finally 96% over 1.9 min, ending at 100% B over 0.1 min. The solvent was held at 100% B for 0.9 min, resulting in a total run time of 12.0 min. Equilibration was initiated by decreasing the solvent from 100% B to 15% B over 0.2 min and maintained for a total of 16 min.

[0201] Data processing was performed using vendor software Qualitative Analysis B.07.00 and QQQ Qualitative Analysis B.10.0.0. Relative quantification of lipid species was determined by comparing them to their corresponding internal standards. Total concentrations of lipid classes were determined by summing the concentrations of individual lipid species within each lipid class.

[0202] statistical analysis Lipidomic analysis was performed on triplicate cell samples. Figures were generated using Microsoft Excel and presented as means with standard deviation error bars. Statistical analysis was performed using GraphPad Prism 9.5.1. First, a one-way analysis of variance (ANOVA) was applied to compare different treatment groups. Then, a post-hoc analysis was performed using Fisher's LSD test to identify significant differences between groups. A P value of less than 0.05 was considered statistically significant.

[0203] Before starting treatment, cells were 2+ and Mg 2+ The cells were washed with phosphate-buffered saline (PBS) containing no ATP. After 24 hours of treatment with precursors, cells were harvested by first washing with PBS, followed by cell scraping. The cell suspension was then dried overnight using an SPD121P SpeedVac concentrator. After drying, the samples were stored at -80°C until lipid extraction was performed.

[0204] result Supplementation with both LPC(O) and LPE(O) resulted in significant increases in total cellular PE(P) levels, albeit to different degrees (7-114% relative to the control group) (Figures 44 and 45). In particular, 100% LPC(O-16:0) demonstrated greater efficacy in elevating endogenous PE(P) levels compared with 100% LPC(O-18:0) and 100% LPC(O-18:1) (Figure 44). Conversely, both 100% LPE(O-16:0) and 100% LPE(O-18:0) demonstrated comparable efficacy in enhancing PE(P) levels, whereas 100% LPE(O-18:1) was relatively less effective (Figure 45). Different combinations of O-16:0, O-18:0, and O-18:1 seeds significantly increased PE(P) levels, but none of the combinations matched the effectiveness of 100% O-16:0 seeds in increasing PE(P) levels (Figures 43-44).

[0205] Different combinations of O-16:0, O-18:0, and O-18:1 species also significantly affected intracellular plasmalogen composition (Figures 46 and 47). Within LPC(O) treatment, both single- and dual-species supplementation resulted in changes in the relative proportions of 16:0, 18:0, and 18:1 PE(P) in cells (Figure 45). However, certain mixtures, particularly LPC(O-16:0) 34.1% + LPC(O-18:0) 41.5% + LPC(O-18:1) 24.4%, were able to maintain these proportions while increasing total PE(P) levels (Figure 46). Among the LPE(O) treatments, only one combination, LPE(O-16:0) 62%, LPE(O-18:0) 23.9%, and LPE(O-18:1) 14.1%, was successful in maintaining the relative proportions of 16:0, 18:0, and 18:1 PE(P) while simultaneously increasing total PE(P) levels (Figure 46), whereas all other combinations increased total PE(P) levels while altering the relative proportions of these species.

[0206] Without wishing to be bound by theory, these results establish the efficacy of the novel plasmalogen precursors LPC(O) and LPE(O) in significantly increasing cellular PE(P) levels in RAW264.7 macrophage cells. Nevertheless, the extent of this increase showed great variability across different SN1 compositions for both LPC(O) and LPE(O) compounds. In particular, O-16:0 was found to be the most potent at increasing endogenous PE(P) levels, compared to O-18:0 and O-18:1 species, at 100%. This disparity highlights the differential effects of precursor species with different SN1 compositions on cellular plasmalogen levels and may be due to variations in their metabolic processes or their distinct effects on the de novo plasmalogen synthesis pathway.

[0207] Without being bound by theory, the prevalence of 16:0 species as the most abundant PE(P) species in these cells suggests a potential preference for accumulating more 16:0 species after supplementation. Notably, alterations in certain PE(P) species are known to have opposite effects on other species. Consequently, increasing 18:0 or 18:1 PE(P) species through supplementation with O-18:0 or O-18:1 precursors, respectively, may result in stronger feedback inhibition of de novo synthesis of 16:0 PE(P) species, resulting in a substantially lower overall increase in total cellular PE(P) levels.

[0208] These findings demonstrate the significant influence of precursor compounds with different SN1 compositions on intracellular plasmalogen composition. The specific mixture of LPC(O) and LPE(O) treatments can increase total PE(P) levels while maintaining the relative proportions of 16:0, 18:0, and 18:1 PE(P) species, demonstrating a potential means of modulating cellular plasmalogen composition without disrupting the balance of individual species. [Table 15-1] [Table 15-2] [Table 15-3] [Table 15-4] [Table 15-5] [Table 15-6] [Table 15-7] [Table 15-8]

[0209] For all transitions, the polarity was positive and the fragmentor 166 and cell accelerator voltages were set to 5.

[0210] LPC: lysophosphatidylcholine; LPC(O): lysoalkylphosphatidylcholine; LPC(P): lysoalkenylphosphatidylcholine; LPE: lysophosphatidylethanolamine; LPE(O): lysoalkylphosphatidylethanolamine; LPE(P): lysoalkenylphosphatidylethanolamine; PC: phosphatidylcholine; PC(O): alkylphosphatidylcholine; PC(P): alkenylphosphatidylcholine; PE: phosphatidylethanolamine; PE(O): alkylphosphatidylethanolamine; PE(P): alkenylphosphatidylethanolamine [PE(P)]; TG: triacylglycerol; TG(O): alkyldiacylglycerol; ISTD: internal standard; NL: neutral loss; SIM: single ion monitoring.

[0211] Example 6 This example relates to the modulation of plasmalogen levels and composition by supplementation of LPC(O) and LPE(O) in 3T3-L1 cells with different SN-1 compositions.

[0212] The methods employed in this study were the same as those described in Example 5, except for the cell line and culture medium used. In this study, the mouse fibroblast / preadipocyte cell line 3T3-L1 was utilized and cultured in Dulbecco's modified Eagle's medium (DMEM) (Gibco, USA) supplemented with 1% sodium pyruvate (Gibco, USA) and 10% newborn calf serum (Gibco, New Zealand).

[0213] In 3T3-L1 preadipocytes, none of the plasmalogen precursor treatments resulted in a significant increase in cellular PE(P) levels; rather, some treatments, particularly 100% LPC(O-18:1) and 100% LPE(O-18:1), appeared to decrease these levels (Figures 47-48). Examination of the effects of both LPC(O) and LPE(O) treatments revealed subtle changes in the relative proportions of 16:0, 18:0, and 18:1 PE(P) with single species supplementation; i.e., supplementation of specific SN1 species increased the proportion of the corresponding PE(P) species but decreased the proportion of other PE(P) species (Figures 49-50).

[0214] In 3T3-L1 cells, no increase in cellular PE(P) levels was observed after supplementation with LPC(O) or LPE(O) under the conditions tested. These cells already exhibit high baseline levels of PE(P), which may explain the lack of further increase with precursor supplementation. Nevertheless, this study revealed a significant effect of precursor compounds with different SN1 compositions on cellular PE(P) composition. Without wishing to be bound by theory, these results emphasize the role of SN1 / alkyl chain composition in plasmalogen precursor supplements to preserve natural plasmalogen composition in healthy subjects.

[0215] Example 7 The murine macrophage cell line RAW264.7 was used in additional studies to determine whether any functional / phenotypic outcomes were measurable as a result of the increased plasmalogen levels / compositional changes observed by feeding cells with LPC(O) and LPE(O) ether lipids.

[0216] The subset of compound mixtures in Table 14 with various SN-1 fatty acids were selected based on ratios that appear to maintain the ratio of plasmalogen compounds most commonly observed in healthy human subjects. The compound mixtures tested were as follows: Vehicle control LPC(O-18:1) 100% LPC(O-16:0)34.1%+LPC(O-18:0)41.5%+LPC(O-18:1)24.4% LPC(O-16:0)62%+LPC(O-18:0)23.9%+LPC(O-18:1)14.1% LPE(O-18:1) 100% LPE(O-16:0)34.1%+LPE(O-18:0)41.5%+LPE(O-18:1)24.4% LPE(O-16:0)62%+LPE(O-18:0)23.9%+LPE(O-18:1)14.1%

[0217] Cells were pretreated with 20 μM of the precursor compound mixture for 24 hours and then treated with LPS (1 nM) for 6 hours to stimulate the immune response. After LPS treatment, cells were harvested for gene expression analysis using qPCR. Total RNA was isolated from the cells using TRIzol™ Reagent (Invitrogen, Thermo Fisher Scientific, USA). 500 μl of TRIzol was added to each well of a cell culture plate, followed by lifting the cells from the plate using a scraper. The resulting cell lysate was stored at -80°C until further processing. On the day of RNA isolation, the cell lysate was thawed, and 100 μl of chloroform was added to each sample. Each sample tube was then vigorously mixed for 20 seconds and then incubated at room temperature for 5 minutes. The samples were then centrifuged at 12,000 g for 15 minutes at 4°C.

[0218] The aqueous phase was carefully transferred to a new tube. 500 μl of isopropanol was added to each sample, mixed thoroughly, and incubated at room temperature for 30 minutes. The samples were then centrifuged at 17,000 g for 15 minutes at 4°C. The supernatant was decanted, leaving the RNA pellet.

[0219] To the RNA pellet, 1 ml of 75% molecular-grade ethanol (Sigma-Aldrich, USA) was added, vortexed briefly for 5 seconds, and centrifuged at 7500 g for 5 minutes at 4°C. The supernatant was then discarded. An additional 1 ml of 75% ice-cold ethanol was then added to the pellet, followed by centrifugation at 7500 g for 5 minutes at 4°C. This washing process was repeated once more, and after the final centrifugation, the supernatant was discarded, leaving the washed RNA pellet.

[0220] The tube containing the pellet was dried on a heating block at 55°C for 10 minutes. Once dry, 20 μl of molecular-grade water (Sigma-Aldrich, USA) was added to dissolve the RNA pellet. The quantity and quality of the isolated RNA were determined using a Nanodrop spectrophotometer (Thermo Fisher Scientific, USA), and the RNA samples were stored at -80°C until further analysis.

[0221] Complementary DNA was synthesized by reverse transcription using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Thermo Fisher Scientific, USA) according to the manufacturer's recommendations.

[0222] Quantitative real-time PCR was performed using Taqman™ assays (Il6: Mm00446190_m1, Nfe2l2: Mm00477784_m1, Tlr4: Mm00445273_m1, Acox1: Mm00443579_m1, Cpt1α: Mm00550438_m1, Sod1: Mm01344233_g1, Pex16: Mm00455021_m1, Hnrnpab: Mm01288699_m1) and TaqMan™ Fast Advanced Master Mix (Applied Biosystems, Thermo Fisher Scientific, USA) and amplified on an Applied Biosystems Quant 7 real-time PCR instrument (Life Technologies, Thermo Fisher Scientific, USA) according to the manufacturer's recommendations. Target gene expression was normalized to the expression of a housekeeping gene [heterogeneous nuclear ribonucleoprotein (Hnrnpab)] and a reference group (control or control + LPS) using the 2-ΔΔCt method of quantification and expressed as a relative value (Livak KJ, Schmittgen TD. Methods. Vol. 25. San Diego, CA: 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method; pp. 402-408.)

[0223] Data are presented as mean ± SD (n = 3 / group). Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. An asterisk (*) indicates P < 0.05 compared to control, and ^ indicates P < 0.05 compared to control + LPS.

[0224] None of the compounds tested appeared to affect levels of TNFα as measured by ELISA (data not shown), although effects were seen at the gene expression level for many indicators of inflammatory / immune response.

[0225] LPC(O-18:1) supplementation, like all LPE(O) treatments, suppressed the LPS-induced increase in IL6 gene expression. All LPX(O) treatments reduced NFE2L2 gene expression, but only four of the six (mixtures 4–7 above, including all LPE(O) treatments) were statistically significant in this assay.

[0226] All LPX(O) treatments suppressed the LPS-induced increase in TLR4 gene expression (Figure 55). Reductions ranged from 11 to 20% compared to the induction level after LPS treatment, minus the vehicle control level.

[0227] These data suggest that there may be anti-inflammatory activity associated with supplementation of LPX(O), especially LPE(O), in RAW264.7 cells. (Xing, X. et al., IL-6 is an anti-inflammatory cytokine required for controlling local or systemic acute inflammatory responses. J Clin Invest. 1998;15:311-320; He, F. et al., NRF2, a transcription factor for stress response and beyond. Int. J. Mol. Sci. 2020:21(13),4777; Saleh et al., The anti-inflammatory properties of phytochemicals and their effects on epigenetic mechanisms involved in TLR4 / NF-kB-mediated inflammation. Front. Immunol. 2021:12)

[0228] RAW264.7 macrophage cells were treated with 20 μM LPC (O) or LPE (O) with different SN1 compositions for 24 hours, followed by LPS (1 nM) for 6 hours, and then harvested for gene expression analysis. Gene expression data were normalized to Hnrnpab expression and presented as mean ± SD (n = 3 / group). Mean differences between groups were compared by one-way ANOVA followed by Fisher's LSD test. The results are shown in Figure 56, where * indicates P < 0.05 compared to the control group and ^ indicates P < 0.05 compared to the control group + LPS.

[0229] Example 8 Formulation of LPX(O) compounds. Certain LPX(O) compound(s) and compositions have been formulated for stable aqueous delivery. Formulation of emulsified LPX(O) compounds that can withstand freeze / thaw processes and / or are suitable for delivery to animals for pharmacokinetic studies has been achieved by mixing the LPX(O) compound(s) and compositions with a solubilizing agent.

[0230] Preparation of alkylglycerol, alkyldiacylglycerol, and lysoalkylphosphatidylcholine formulations. 1. Stock solutions (100 mg / ml) of the individual precursor compounds were prepared in a chloroform:methanol (1:1) solution. 2. A stock solution (100 mg / ml) of purified egg yolk L-α-lecithin (Sigma-Aldrich, St. Louis, MO, USA) was similarly prepared in a chloroform:methanol (1:1) solution. 3. Precursor compound mixtures were prepared by combining appropriate volumes of precursor compound stocks (Table 15). 4. The mixture of precursor compounds and lecithin stock was then dispensed into scintillation vials (Table 15). 5. The mixture of precursor compound and lecithin was then dried at 40°C under a stream of nitrogen gas. 6. After complete evaporation of chloroform and methanol, the dried lipids were reconstituted in 3 ml of deionized water by vigorous vortexing, followed by sonication for 1 h in an ultrasonic cleaner water bath (Soniclean, Adelaide, South Australia, Australia) and further sonication for 2 × 30 s each at an amplitude of 25 using a Misonix S-4000 sonicator (Thermo Fisher Scientific, Melbourne, Victoria, Australia). [Table 16]

[0231] Preparation of lysoalkylphosphatidylethanolamine formulations 1. Stock solutions (10 mg / ml) of the individual precursor compounds were prepared in chloroform:methanol (2:1) solution*. 2. A stock solution (100 mg / ml) of purified egg yolk L-α-lecithin (Sigma-Aldrich, St. Louis, MO, USA) was similarly prepared in a chloroform:methanol (2:1) solution. 3. Precursor compound mixtures were prepared by combining appropriate volumes of precursor compound stocks (Table 16). 4. The mixture of precursor compounds and lecithin stock was then dispensed into glass tubes (Table 16). 5. The mixture of precursor compound and lecithin was then dried at 40°C under a stream of nitrogen gas. 6. After complete evaporation of chloroform and methanol, the dried lipids were reconstituted in 3 ml of deionized water by vigorous vortexing, followed by sonication for 1 h in an ultrasonic cleaner water bath (Soniclean, Adelaide, South Australia, Australia) and further sonication for 2 × 30 s each at an amplitude of 25 using a Misonix S-4000 sonicator (Thermo Fisher Scientific, Melbourne, Victoria, Australia). *LPE(O-16:0), LPE(O-16:0)-d5, and LPE(O-18:0) solutions were briefly heated in a water bath at approximately 40°C. [Table 17]

[0232] Without wishing to be bound by theory, it is believed that analysis of the above-described formulations indicates that purified egg yolk L-α-lecithin is a suitable solubilizing agent for use in preparing stable aqueous formulations that are suitable for delivery to animals and that can withstand freezing and thawing, for example. Without wishing to be bound by theory, it is believed that formulations that include a solubilizing agent such as purified egg yolk L-α-lecithin may have improved stability and / or better resistance to freezing and thawing, e.g., by being less susceptible to degradation during freezing and thawing, compared to similar formulations that do not include the solubilizing agent.

[0233] Given the improved bioavailability of LPC(O), various combinations of LPC(O), LPE(O), and LPA(O) are being considered for use in pharmaceutical formulations and dietary supplements. Dosages and serving sizes may vary depending on the subject's age, height, and weight.

[0234] In one embodiment, a useful dose for humans is 2-3 mg / kg per person per day, which corresponds to 100-300 mg of LPC(O) equivalents. In a further embodiment, a useful dose for humans is 200 mg of LPC(O) equivalents per person per day.

[0235] In another embodiment, the maintenance dose may be 0.2-2 mg / kg, or 25-100 mg of LPC(O) equivalent per person per day, i.e., depending on the individual, the maintenance dose may be 25 mg, 50 mg, or 100 mg of LPC(O) equivalent per person per day.

[0236] In another embodiment, if a large or rapid increase in plasmalogens is desired, the dose can be 3-25 mg / kg, 300-2000 mg LPC(O) equivalents per person per day, i.e., if a large or rapid increase in plasmalogens is desired, the dose can be 400 mg, 800 mg, or 1600 mg LPC(O) equivalents per person per day.

[0237] In another embodiment, the dose may consist of 0.1 to 4000 mg of LPC(O) equivalent per person per day.

[0238] The use of the terms "a," "an," "the," and similar referents in the context of describing the present disclosure (particularly in the context of the claims) should be construed to encompass both the singular and the plural unless otherwise indicated herein or clearly contradicted by context. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated herein as if set forth individually herein. The use of the term "about" is intended to describe a value above or below the stated value by about ±10%; in other embodiments, the value may range by about ±5% above or below the stated value. In other embodiments, the value may range by about ±2% above or below the stated value. In other embodiments, the value may range by about ±1% above or below the stated value. The foregoing ranges are intended to be made clear by context, and no further limitation is implied. All methods described herein can be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "etc.") provided herein is intended merely to better clarify the disclosure and does not limit the scope of the disclosure unless otherwise stated. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0239] In the foregoing specification, while the present disclosure has been described with reference to specific embodiments thereof, and numerous details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the present disclosure is susceptible to additional embodiments, and that particular of the details described herein can be varied considerably without departing from the underlying principles of the present disclosure.

[0240] All references cited herein are incorporated by reference in their entirety. The present disclosure may be embodied in other specific forms without departing from its spirit or essential attributes, and therefore, reference should be made to the appended claims, rather than the foregoing specification, as indicating the scope of the present disclosure.

Claims

1. At least one compound of formula (I) or a pharmaceutically acceptable salt thereof 【Chemistry 1】 A composition comprising:

2. R ３ The composition of claim 1 , wherein is selected from phosphates and substituted phosphates.

3. R ３ The composition of claim 2 , wherein is a substituted phosphate.

4. 4. The composition of claim 3, wherein the substituted phosphate is substituted with an alkylamine or an inositol.

5. The alkylamine is 【Chemistry 2】 and 【Transformation 3】 The composition of claim 4, wherein the composition is selected from:

6. R １ and R ２ are each independently hydrogen, optionally substituted C １－３０ alkyl group, optionally substituted C ２－３０ Alkenyl groups and optionally substituted C １－３０ The composition according to any one of claims 1 to 5, wherein the hydroxyl group is selected from acyl groups.

7. R １ and R ２ each independently represents an optionally substituted C １４－２４ alkyl group, optionally substituted C １４－２４ Alkenyl groups and optionally substituted C １４－２４ The composition of claim 6, wherein the hydroxyl group is selected from acyl groups.

8. R ２ is hydrogen, R １ is an optionally substituted C １－３０ alkyl group, optionally substituted C ２－３０ alkenyl group, or optionally substituted C １－３０ The composition of claim 6, wherein the group is an acyl group.

9. R １ is an optionally substituted C １４－２４ alkyl group, optionally substituted C １４－２４ alkenyl group, or optionally substituted C １４－２４ The composition of claim 8, wherein the group is an acyl group.

10. R １ is an optionally substituted C １４－１８ alkyl group, optionally substituted C １４－１８ alkenyl group, or optionally substituted C １４－１８ The composition of claim 9, wherein the group is an acyl group.

11. R １ is unsubstituted C １６ The composition of claim 10, wherein the group is an alkyl group.

12. R １ is unsubstituted C １８ The composition of claim 10, wherein the group is an alkyl group.

13. R １ is unsubstituted C １８ The composition of claim 10, wherein the alkyl group is an alkenyl group.

14. The at least one compound of formula (I) is a compound of formula (IA) or a pharmaceutically acceptable salt thereof 【Chemistry 4】 2. The composition of claim 1, wherein:

15. Each R ｘ is hydrogen or C １－３ 15. The composition of claim 14, wherein the alkyl is independently selected from alkyl.

16. 16. The composition of claim 14 or 15, wherein n is 2 or 3.

17. R ｘ 15. The composition of claim 14, wherein is methyl and n is 3.

18. R Ａ The composition of any one of claims 14 to 17, wherein is an optionally substituted hydrocarbon chain containing from 14 to 24 carbon atoms.

19. R Ａ 19. The composition of claim 18, wherein is an optionally substituted hydrocarbon chain containing 16 to 18 carbon atoms.

20. R Ａ is an optionally substituted C １６－１８ alkyl group, optionally substituted C １６－１８ alkenyl group, or optionally substituted C １６－１８ 20. The composition of claim 19, wherein the group is an acyl group.

21. R Ａ is an optionally substituted C １６－１８ The composition of claim 20, wherein the group is an alkyl group.

22. R Ａ is an optionally substituted C １６－１８ The composition of claim 20, wherein the alkyl group is an alkenyl group.

23. R Ａ is unsubstituted C １６ 20. The composition of claim 19, wherein the group is an alkyl group.

24. R Ａ is unsubstituted C １８ 20. The composition of claim 19, wherein the group is an alkyl group.

25. R Ａ is unsubstituted C １８ The composition of claim 19, wherein the alkyl group is an alkenyl group.

26. The composition according to any of claims 1 to 25, wherein the composition comprises a mixture of at least two compounds of formula (I) and / or (IA).

27. The composition of any of claims 1 to 26, wherein the composition comprises a mixture of three compounds of formula (I) and / or (IA).

28. The compound of formula (IA) has the structure (IA1), (IA2), or (IA3): 【Transformation 5】 15. The composition of claim 14, wherein

29. 29. The composition of claim 28, wherein the composition comprises a mixture of at least two compounds selected from the group consisting of (I-A1), (I-A2), and (I-A3).

30. 29. The composition of claim 28, wherein the composition comprises a mixture of (I-A1), (I-A2), and (I-A3).

31. The composition of claim 28, wherein the composition comprises a mixture of (I-A1), (I-A2), and (I-A3), and the mixture of (I-A1), (I-A2), and (I-A3) accounts for at least 50% of the ether lipids in the composition on a molar percentage basis.

32. 29. The composition of claim 28, wherein the mixture has a molar ratio of (I-A2) to (I-A1) of 1.2:1 to 2.5:

1.

33. 33. The composition of claim 32, wherein the mixture has a molar ratio of (I-A2) to (I-A1) of 1.5:1 to 2.1:

1.

34. 34. The composition of claim 33, wherein the mixture has a molar ratio of (I-A2) to (I-A1) of 1.7:

1.

35. 29. The composition of claim 28, wherein the mixture has a molar ratio of (I-A2) to (I-A3) of 0.9:1 to 1.7:

1.

36. 36. The composition of claim 35, wherein the mixture has a molar ratio of (I-A2) to (I-A3) of 1:1 to 1.5:

1.

37. 37. The composition of claim 36, wherein the mixture has a molar ratio of (I-A2) to (I-A3) of 1.22:

1.

38. 29. The composition of claim 28, wherein the mixture has a molar ratio of (I-A1) to (I-A3) of 0.5:1 to 1:

1.

39. 39. The composition of claim 38, wherein the mixture has a molar ratio of (I-A1) to (I-A3) of 0.6:1 to 0.9:

1.

40. 40. The composition of claim 39, wherein the mixture has a molar ratio of (I-A1) to (I-A3) of 0.72:

1.

41. 31. The composition of claim 30, wherein the mixture has a molar percentage of (I-A1) of 18.6% to 27.9%, a molar percentage of (I-A2) of 32.6% to 45.8%, and a molar percentage of (I-A3) of 26.8% to 37.4%.

42. 31. The composition of claim 30, wherein the mixture has a molar percentage of (I-A1) of 23.3%, a molar percentage of (I-A2) of 39.2%, and a molar percentage of (I-A3) of 32.1%.

43. 31. The composition of claim 30, wherein the mixture has a molar ratio of (I-A1):(I-A2):(I-A3) of 1:1.7:1.

4.

44. The compound of formula (IA) has the structure (I-A4), (I-A5), or (I-A6): 【Transformation 6】 15. The composition of claim 14, wherein

45. 45. The composition of claim 44, wherein the composition comprises a mixture of at least two compounds selected from the group consisting of (I-A4), (I-A5), and (I-A6).

46. 45. The composition of claim 44, wherein the composition comprises a mixture of (I-A4), (I-A5), and (I-A6).

47. The composition of claim 44, wherein the composition comprises a mixture of (I-A4), (I-A5), and (I-A6), and the mixture of (I-A4), (I-A5), and (I-A6) accounts for at least 50% of the ether lipids in the composition on a molar percentage basis.

48. 45. The composition of claim 44, wherein the mixture has a molar ratio of (I-A5) to (I-A4) of 1.2:1 to 2.5:

1.

49. 49. The composition of claim 48, wherein the mixture has a molar ratio of (I-A5) to (I-A4) of 1.5:1 to 2.1:

1.

50. 50. The composition of claim 49, wherein the mixture has a molar ratio of (I-A5) to (I-A4) of 1.7:

1.

51. 45. The composition of claim 44, wherein the mixture has a molar ratio of (I-A5) to (I-A6) of 0.9:1 to 1.7:

1.

52. 52. The composition of claim 51, wherein the mixture has a molar ratio of (I-A5) to (I-A6) of 1:1 to 1.5:

1.

53. 53. The composition of claim 52, wherein the mixture has a molar ratio of (I-A5) to (I-A6) of 1.29:

1.

54. 45. The composition of claim 44, wherein the mixture has a molar ratio of (I-A4) to (I-A6) of 0.5:1 to 1:

1.

55. 55. The composition of claim 54, wherein the mixture has a molar ratio of (I-A4) to (I-A6) of 0.6:1 to 0.9:

1.

56. 56. The composition of claim 55, wherein the mixture has a molar ratio of (I-A4) to (I-A6) of 0.76:

1.

57. 47. The composition of claim 46, wherein the mixture has a molar percentage of (I-A4) between 18.6% and 27.9%, a molar percentage of (I-A5) between 32.6% and 45.8%, and a molar percentage of (I-A6) between 26.8% and 37.4%.

58. 58. The composition of claim 57, wherein the mixture has a molar percentage of (I-A4) of 24.8%, a molar percentage of (I-A5) of 42.4%, and a molar percentage of (I-A6) of 32.8%.

59. 47. The composition of claim 46, wherein the mixture has a molar ratio of (I-A4):(I-A5):(I-A6) of 1:1.7:1.

3.

60. A formulation, such as a pharmaceutical formulation, comprising a composition as defined in any one of claims 1 to 59 and at least one excipient.

61. 61. The formulation of claim 60, wherein the formulation is formulated for oral administration.

62. 62. The formulation of claim 61, wherein the oral dosage form is a tablet, capsule, solution, mouthwash, suspension, powder, gum, confectionery, lozenge, sublingual delivery system, or fast dissolving formulation.

63. The formulation of any one of claims 60 to 62, wherein the formulation is a food product.

64. 64. The formulation of claim 63, wherein the formulation is a dietary supplement.

65. 64. The formulation of claim 63, wherein the food is a medical food.

66. 64. The formulation of claim 63, wherein the food product is an infant formula.

67. The formulation of any one of claims 60 to 66, for maintaining or regulating plasmalogen levels in a human subject in need thereof.

68. The formulation of any one of claims 60 to 66, for maintaining or regulating plasmalogen levels in an animal subject in need thereof.

69. 69. The formulation of claim 68, wherein the animal is selected from domestic animals, working animals and farm animals.

70. The formulation of any one of claims 67 to 69, wherein the maintaining or regulating requires maintaining or regulating the level of plasmalogens at levels and / or ratios associated with a non-disease state.

71. A composition as defined in any one of claims 1 to 59, or a formulation as defined in any one of claims 60 to 70, for use in increasing the level of a plasmalogen compound in the blood or tissue of a subject.

72. 72. The composition for use of claim 71, wherein the subject is a human.

73. 72. The composition for use of claim 71, wherein the subject is an animal.

74. 74. The composition for use according to claim 73, wherein the animal subject is selected from domestic animals, working animals and farm animals.

75. A composition as defined in any one of claims 1 to 59 or a formulation as defined in any one of claims 60 to 70 for use in therapy.

76. A composition as defined in any one of claims 1 to 59, or a formulation as defined in any one of claims 60 to 70, for use in the treatment of a disease or disorder associated with a deficiency of plasmalogens.

77. 77. The composition or formulation for use according to claim 76, wherein the disease or disorder is a neurological disease.

78. 78. The composition or formulation for use of claim 77, wherein the neurological disease is selected from Alzheimer's disease, Parkinson's disease, autism, amyotrophic lateral sclerosis, frontotemporal dementia, multiple sclerosis, and schizophrenia.

79. 77. The composition or formulation for use according to claim 76, wherein the disease or disorder is a metabolic disorder.

80. 80. The composition or formulation of claim 79, wherein the metabolic disorder is selected from the group consisting of obesity, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, immune-related diseases, cardiovascular diseases, neurological diseases, cancer, myalgic encephalomyelitis / chronic fatigue syndrome, Barth syndrome, and peroxisomal disorders.

81. 81. The composition or formulation of claim 80, wherein the immune-related disease is selected from the group consisting of asthma, atopic dermatitis, type 1 diabetes, and infectious diseases.

82. 81. The composition or formulation of claim 80, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cardiac remodeling, and hypertension.

83. 81. The composition or formulation of claim 80, wherein the peroxisomal disorder is Zellweger syndrome spectrum disorder or rhizomelic chondrodysplasia punctata.

84. 84. The composition or formulation for use according to any one of claims 71 to 83, wherein the compound of formula (I) or (IA) is administered in a dose of 0.1 to 4000 mg per day.

85. 85. The composition or formulation for use according to claim 84, wherein the compound of formula (I) or (IA) is administered at a dose of 0.1 to 2000 mg per day.

86. 86. The composition or formulation for use according to claim 85, wherein the compound of formula (I) or (IA) is administered at a dose of 25 to 1600 mg per day.

87. 87. The composition or formulation for use of claim 86, wherein the compound of formula (I) or (IA) is administered at a dose of 400 mg, 800 mg, or 1600 mg per day.

88. 87. The composition or formulation for use of claim 86, wherein the compound of formula (I) or (IA) is administered at a dose of 200 mg per day.

89. 87. The composition or formulation for use according to claim 86, wherein the compound of formula (I) or (IA) is administered at a dose of 25 to 100 mg per day.

90. A method for increasing the level of a plasmalogen compound in the blood or tissue of a subject in need thereof, comprising administering to the subject an effective amount of at least one compound of formula (I) or a pharmaceutically acceptable salt thereof, and / or at least one compound of formula (IA) or a pharmaceutically acceptable salt thereof, as defined in claims 1 and 14, respectively.

91. 91. The method of claim 90, wherein the subject is a human.

92. 91. The method of claim 90, wherein the subject is an animal.

93. 93. The method of claim 92, wherein the animal subject is selected from domestic animals, working animals, and farm animals.

94. The method of claim 90, wherein the bioavailability of the plasmalogen compound in the blood or tissues of the subject is improved after administration of at least one compound of formula (I) or a pharmaceutically acceptable salt thereof, and / or at least one compound of formula (I-A) or a pharmaceutically acceptable salt thereof, compared to administration of AKG (alkylglycerol).

95. A method for treating a disease or disorder associated with a deficiency of plasmalogens, comprising administering to a subject in need thereof an effective amount of at least one compound of formula (I) or a pharmaceutically acceptable salt thereof, and / or at least one compound of formula (IA) or a pharmaceutically acceptable salt thereof, as defined in claims 1 and 14, respectively.

96. 96. The method of claim 95, wherein the subject is a human.

97. 96. The method of claim 95, wherein the subject is an animal.

98. 98. The method of claim 97, wherein the animal subject is selected from domestic animals, working animals, and farm animals.

99. 96. The method of claim 95, wherein the disease or disorder is a neurological disease.

100. 100. The method of claim 99, wherein the neurological disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, autism, amyotrophic lateral sclerosis, frontotemporal dementia, multiple sclerosis, and schizophrenia.

101. 96. The method of claim 95, wherein the disease or disorder is a metabolic disorder.

102. 102. The method of claim 101, wherein the metabolic disorder is selected from the group consisting of obesity, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, immune-related diseases, cardiovascular diseases, neurological diseases, cancer, myalgic encephalomyelitis / chronic fatigue syndrome, Barth syndrome, and peroxisomal disorders.

103. 103. The method of claim 102, wherein the immune-related disease is selected from the group consisting of asthma, atopic dermatitis, type 1 diabetes, and infectious diseases.

104. 103. The method of claim 102, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cardiac remodeling, and hypertension.

105. 103. The method of claim 102, wherein the peroxisomal disorder is Zellweger syndrome spectrum disorder or rhizomelic chondrodysplasia punctata.

106. 106. The method of any one of claims 90 to 105, wherein the compound of formula (I) or (IA) is administered at a dose of 0.1 to 4000 mg per day.

107. 107. The method of claim 106, wherein the compound of formula (I) or (IA) is administered at a dose of 0.1 to 2000 mg per day.

108. 108. The method of claim 107, wherein the compound of formula (I) or (IA) is administered at a dose of 25 to 1600 mg per day.

109. 109. The method of claim 108, wherein the compound of formula (I) or (IA) is administered at a dose of 400 mg, 800 mg, or 1600 mg per day.

110. 109. The method of claim 108, wherein the compound of formula (I) or (IA) is administered at a dose of 200 mg per day.

111. 109. The method of claim 108, wherein the compound of formula (I) or (IA) is administered at a dose of 25 to 100 mg per day.

112. 112. The method of claim 111, wherein the compound of formula (I) or (IA) is administered at a dose of 25 mg, 50 mg, or 100 mg per day.

113. Use of a composition according to any one of claims 1 to 59 or a formulation according to any one of claims 60 to 70 in the manufacture of a medicament for increasing the level of a plasmalogen compound in the blood or tissue of a subject in need thereof.

114. Use of a composition according to any one of claims 1 to 59 or a formulation according to any one of claims 60 to 70 in the manufacture of a medicament for treating a disease or disorder associated with a deficiency of plasmalogens.

115. 115. The use of claim 114, wherein the disease or disorder is a neurological disease.

116. 116. The use of claim 115, wherein the neurological disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, autism, amyotrophic lateral sclerosis, frontotemporal dementia, multiple sclerosis, and schizophrenia.

117. 117. The use of claim 116, wherein the disease or disorder is a metabolic disorder.

118. 118. The use of claim 117, wherein the metabolic disorder is selected from the group consisting of obesity, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, immune-related diseases, cardiovascular diseases, neurological diseases, cancer, myalgic encephalomyelitis / chronic fatigue syndrome, Barth syndrome, and peroxisomal disorders.

119. 119. The use of claim 118, wherein the immune-related disease is selected from the group consisting of asthma, atopic dermatitis, type 1 diabetes, and infectious diseases.

120. 119. The use of claim 118, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cardiac remodeling, and hypertension.

121. 119. The use of claim 118, wherein the peroxisomal disorder is Zellweger syndrome spectrum disorder or rhizomelic chondrodysplasia punctata.

122. 113. A kit for use in the method of any one of claims 90 to 112, comprising at least one compound of formula (I) or formula (IA), or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 and 14, respectively.

123. 29. The composition of claim 28, wherein the at least one compound is selected from compounds of formula (I-A1), (I-A2), or (I-A3).

124. 30. The composition of claim 29, wherein the mixture of at least two compounds is a 50:50 mixture.

125. The mixture of (I-A1), (I-A2) and (I-A3) is LPC (O-16:0) 26.8% + LPC (O-18:0) 46.1% + LPC (O-18:1) 27.1%, LPC (O-16:0) 46% + LPC (O-18:0) 21% + LPC (O-18:1) 33%, LPC (O-16:0) 42% + LPC (O-18:0) 51% + LPC (O-18:1) 7%, LPC (O-16:0) 62% + LPC (O-18:0) 23.9% + LPC (O-18:1) 14.1%, LPC (O-16:0) 19.8% + LPC (O-18:0) 66% + LPC (O-18:1) 14.2%, 32.5% LPC (O-16:0) + 39.6% LPC (O-18:0) + 27.9% LPC (O-18:1), or The composition according to claim 30, selected from 34.1% LPC (O-16:0) + 41.5% LPC (O-18:0) + 24.4% LPC (O-18:1).

126. 45. The composition of claim 44, wherein the at least one compound is selected from compounds of formula (I-A4), (I-A5), or (I-A6).

127. 46. ​​The composition of claim 45, wherein the mixture of at least two compounds is a 50:50 mixture.

128. The mixture of (I-A4), (I-A5) and (I-A6) is LPE (O-16:0) 26.8% + LPE (O-18:0) 46.1% + LPE (O-18:1) 27.1%, LPE (O-16:0) 46% + LPE (O-18:0) 21% + LPE (O-18:1) 33%, LPE (O-16:0) 42% + LPE (O-18:0) 51% + LPE (O-18:1) 7%, LPE (O-16:0) 62% + LPE (O-18:0) 23.9% + LPE (O-18:1) 14.1%, LPE (O-16:0) 19.8% + LPE (O-18:0) 66% + LPE (O-18:1) 14.2%, LPE (O-16:0) 32.5% + LPE (O-18:0) 39.6% + LPE (O-18:1) 27.9%, or 47. The composition of claim 46, wherein the LPE(O-16:0) is 34.1% + LPE(O-18:0) is 41.5% + LPE(O-18:1) is 24.4%.

129. The composition of any one of claims 1 to 59 or 123 to 128, wherein the composition maintains or modulates the plasmalogen compound ratio observed in healthy human subjects.

130. 129. The composition of any one of claims 1 to 59 or 123 to 128, wherein the composition results in a reduction of inflammation or an improvement or reduction of symptoms associated with an inflammatory disease.

131. The composition of claim 130, wherein the reduction in inflammation is associated with a reduction in the level of inflammatory cytokines.

132. 129. The composition of any one of claims 1-59 or 123-128, wherein the composition maintains or modulates levels of inflammatory cytokines at levels observed in healthy human subjects.

133. 133. The composition of claim 132, wherein the inflammatory cytokines that are maintained or regulated are selected from IL-6, NFE2L2, and TLR4.

134. A formulation comprising the composition of any one of claims 1 to 59 or 123 to 128 and at least one excipient and / or solubilizer.

135. 135. The formulation of claim 134, wherein the formulation is formulated for oral administration.

136. 136. The formulation of claim 135, wherein the oral dosage form is a beverage or a food product.

137. 136. The formulation of claim 135, wherein the oral dosage form is a food product.

138. 136. The formulation of claim 135, wherein the oral dosage form is an infant formula.

139. 135. The formulation of claim 134, wherein the formulation comprises a solubilizing agent.

140. 140. The formulation of claim 139, wherein the solubilizing agent is selected from the group consisting of sodium carboxymethylcellulose, hypromellose, proline, xanthan gum, maltodextrin, alginate, wax, lipid, oil, alcohol, sugar, microcrystalline cellulose, starch, calcium phosphate, mannitol, sorbitol, erythritol, food grade solvents, phospholipids, DMSO, ethanol, ethyl acetate, and isopropanol.

141. 140. The formulation of claim 139, wherein the solubilizing agent is lecithin.

142. 142. The formulation of claim 141, wherein the lecithin is purified egg yolk L-alpha-lecithin.

143. 140. The formulation of claim 139, wherein the formulation has improved stability compared to a formulation comprising a composition as defined in any one of claims 1 to 59 or 123 to 128 that does not contain a solubilising agent.

144. 140. The formulation of claim 139, wherein the formulation is less susceptible to degradation during freezing and thawing compared to a formulation comprising a composition as defined in any one of claims 1 to 59 or 123 to 128 that does not contain a solubilising agent.

145. 45. The composition defined in any one of claims 28 or 44, wherein the composition comprises one compound having a purity of at least 99.9%.

146. 146. A formulation comprising the composition defined in claim 145 and at least one excipient.

147. 147. A formulation as defined in any one of claims 134 to 138 or claim 146, wherein the formulation comprises at least one of a solubilizer, emulsifier, stabilizer, dispersant, antifoaming agent, or diluent.

148. 148. A formulation as defined in any one of claims 134 to 138 or claims 146 to 147, wherein the formulation further comprises at least one solubilising agent.

149. The formulation of claim 148, wherein the solubilizing agent is selected from the group consisting of sodium carboxymethylcellulose, hypromellose, proline, xanthan gum, maltodextrin, alginate, wax, lipid, oil, alcohol, sugar, microcrystalline cellulose, starch, calcium phosphate, mannitol, sorbitol, erythritol, food grade solvents, phospholipids, DMSO, ethanol, ethyl acetate, and isopropanol.

150. 150. The formulation of claim 149, wherein the solubilizing agent is selected from the group consisting of DMSO, ethanol, ethyl acetate, and isopropanol.

151. 151. A formulation as defined in any one of claims 134 to 144 or claims 146 to 150, wherein the formulation further comprises at least one antioxidant compound.

152. 152. A formulation as defined in any one of claims 134 to 144 or claims 146 to 151, wherein the formulation further comprises at least one antifoaming agent.

153. 153. A formulation as defined in any one of claims 134 to 144 or claims 146 to 152, wherein the formulation is formulated for oral administration.

154. 154. The formulation of claim 153, wherein the oral dosage form is a tablet, capsule, solution, mouthwash, suspension, powder, gum, confectionery, lozenge, sublingual delivery system, or fast-dissolving formulation.

155. 155. The formulation of claim 154, wherein the oral dosage form is a tablet.

156. 31. The composition of claim 30, wherein the mixture of (I-A1), (I-A2) and (I-A3) comprises 90% w / v of (I-A1).

157. 31. The composition of claim 30, wherein the mixture of (I-A1), (I-A2) and (I-A3) comprises 90% w / v of (I-A2).

158. 31. The composition of claim 30, wherein the mixture of (I-A1), (I-A2) and (I-A3) comprises 90% w / v of (I-A3).

159. 47. The composition of claim 46, wherein the mixture of (I-A4), (I-A5) and (I-A6) comprises 90% w / v of (I-A4).

160. 47. The composition of claim 46, wherein the mixture of (I-A4), (I-A5) and (I-A6) comprises 90% w / v (I-A5).

161. 47. The composition of claim 46, wherein the mixture of (I-A4), (I-A5) and (I-A6) comprises 90% w / v of (I-A6).

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