Lipid emulsion for parenteral nutrition containing GPC

Abstract

The present disclosure relates to parenteral nutrition preparations including immediately usable parenteral nutrition preparations. More particularly, the present disclosure relates to lipid preparations or emulsions and multi-chamber containers containing the same, wherein the lipid emulsion contains glycerophosphorylcholine as a source of choline. The present disclosure further relates to a method for providing choline to a patient requiring parenteral nutrition and a method for avoiding and / or treating choline deficiency and liver damage.

Description

Technology Field

[0001] The present disclosure relates to parenteral nutrition preparations including immediately usable parenteral nutrition preparations. More particularly, the present disclosure relates to lipid preparations or emulsions and multi-chamber containers containing the same, wherein the lipid emulsion contains glycerophosphorylcholine as a source of choline. The present disclosure further relates to a method for providing choline to a patient requiring parenteral nutrition and a method for avoiding and / or treating choline deficiency and liver damage. Background Technology

[0002] It is known that choline deficiency is a problem, particularly in patients receiving parenteral nutrition, because the endogenous production of choline from parenterally supplied methionine is insufficient (Buchman, Gastroenterology 137 (5), 2009, S119-S128). There is evidence that hepatic steatosis develops during parenteral nutrition as a result of this choline deficiency. Additionally, memory and cognitive deficits and skeletal abnormalities have been described. Given that choline is critical in the synthesis of acetylcholine, the most relevant neurotransmitter in cholinergic cells, choline deficiency may be linked to these memory deficits. Choline deficiency may also play a role in cellular apoptosis.

[0003] Therefore, choline is considered an essential nutrient, and the adequate intake (AI) has been defined as 550 mg / day (~8 mg / kg / day) for adult men and 425 mg / day for women. The acceptable upper limit (UL) for choline intake is 3.5 g / day (~50 mg / kg / day) for adults. The normal plasma-free choline concentration in average adults is 10 to 15 nmol / mL. For infants aged 0 to 6 months with a reference body weight of 7 kg, the adequate AI is approximately 18 mg / kg / day (0.17 mmol / kg / day or 125 mg / day of choline), which is significantly higher than that of adults. For infants aged 7–12 months, the appropriate AI is assumed to be 17 mg / kg / day, which corresponds to 150 mg / day of choline (Dietary Reference Intakes for Thiamin, Riboflavin, Niacin, Vitamin B6, Folate, Vitamin B12, Pantothenic Acid, Biotin, and Choline; Institute of Medicine (US) Standing Committee on the Scientific Evaluation of Dietary Reference Intakes and its Panel on Folate, Other B Vitamins, and Choline. Washington (DC): National Academies Press (US); 1998, Chapter 12).

[0004] Pediatric patients appear to be particularly susceptible to choline deficiency. Choline is transported across the placenta against a concentration gradient by mammary gland epithelial cells, and neonates have some significantly higher plasma-free concentrations of choline compared to adults. This may reflect the importance of choline in child development, possibly brain development (Zeisel, The Journal of Pediatrics, 149 (5), Supplement, 2006, S131-S136). Additionally, choline is secreted in breast milk. Therefore, as choline may be required in significant amounts for phospholipid synthesis in the central nervous system, kidneys, liver, lungs, and skeletal muscle, parenteral formulations for preterm and neonates requiring parenteral nutrition must provide an effective source of choline. Recommendations for choline supplementation for preterm infants currently range widely from 8 to 55 mg / kg / day (Bernhard et al., Eur J Nutr (2018). https: / / doi.org / 10.1007 / s00394-018-1834-7).

[0005] Choline is available as a dietary supplement as choline chloride or choline bitartrate, as well as as lecithin. In food, choline exists in esterified forms such as phosphocholine, glycerophosphocholine, phosphatidylcholine, and sphingomyelin, as well as in a free form. Phosphocholine constitutes approximately 40% of the total choline in human breast milk. α-glycerophosphocholine constitutes an additional approximately 40% of the total choline in human breast milk. Orally ingested nutrients are generally readily absorbed through the intestines and delivered directly to the liver. In plasma, choline is a major water-soluble compound, but the concentrations of phosphocholine and α-glycerophosphocholine are very low. The average plasma concentration of α-glycerophosphocholine is only about 0.42 μmol / L (Bernhard et al. 2018), and therefore, little focus has been placed on this compound in the past.

[0006] At the same time, despite the provision of choline precursor methionine in PN solutions and despite the fact that plasma methionine concentrations were found to be generally high to normal levels in PN patients, plasma-free choline concentrations were found to be significantly lower than normal levels in said patients (Compher et al., Journal of Parenteral and Enteral Nutrition 26 (1), 2002, 57). Phosphatidylcholine contained in lipid emulsions used in parenteral nutrition is also not converted to choline to any significant degree.

[0007] Free choline is not currently included in total parenteral nutrition. Instead, as free choline is the major water-soluble component in plasma and chloride is the major anion, choline chloride is currently considered recommended for parenteral nutrition for patients at a dose of about 25 mg / kg / day (Bernhard et al., 2018). Another source of choline is, for example, CDP choline (sodium salt).

[0008] However, during investigations conducted by the inventors, it was found that choline chloride may not be optimal as currently believed in the industry, partly due to the difficulty of stably introducing choline chloride into lipid emulsions for parenteral nutrition. Additionally, there is some evidence that the presence of choline chloride may lead to the degradation of vitamins, particularly vitamins B1 and B12 (Coelho, 2002: Vitamin stability in premixes and feeds. A practical approach in ruminant diets. Proceedings of the 13th Annual Florida Ruminant Nutrition Symposium, Gainesville, FL, January 10-11, 127-145; Frye, 1994: The performance of vitamins in multicomponent premixes. Proc. Roche Technical Symposium, Jefferson, Georgia).

[0009] In the present invention, it has been found that choline derivatives, glycerophosphocholine of formula (I) (GPC), (2-{[(2S)-2,3-dihydroxypropylphosphonato]oxy}ethyl)trimethylazanium (also referred to as glycerophosphorylcholine), α-glycerophosphocholine, L-alpha-glycerylphosphorylcholine, choline alfoscerate, or α-glycerophosphocholine are choline derivatives that can be advantageously used in parenteral formulations as efficient sources of choline for patients receiving parenteral nutrition. GPC is a water-soluble molecule and is contained in small amounts in various foods, such as red meat, but the content is generally somewhat low.

[0010]

[0011] Most GPCs are synthetically produced from natural phospholipids, such as those found in egg yolks or soybeans. For example, GPC is obtained from purified natural phosphatidylcholine (PC), which is hydrolyzed to remove fatty acids. Subsequently, GPC is further re-acylated and / or treated with specific enzymes to synthesize specific chemical molecules (van Hoogevest and Wendel, Eur. J. Lipid Sci. Technol. 2014, 116, 1088-1107).

[0012] GPC has not been considered until now as a component in lipid emulsions and as a source of choline for use in parenteral nutrition.

[0013] US 8865641 B2 discloses a method for treating fatty liver disease in a subject, comprising administering an effective amount of a cholinergic pathway stimulator to the subject, wherein the nicotinic receptor agonist may be choline. However, GPC and / or lipid emulsions for parenteral nutrition are not mentioned.

[0014] US 8337835 B2 describes a method for treating peroxisome proliferator-activated receptor alpha (PPARα)-related liver disorder in mammals, which reduces triglyceride levels and / or increases high-density lipoprotein levels, said method comprising administering a therapeutically effective amount of 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (16:0 / 18:1-GPC) to said mammals.

[0015] US 20110015154 A1 discloses a composition and kit containing a choline compound and a carnitine compound and a method of using the composition and kit, wherein the choline compound may be alpha-GPC. However, US 20110015154 A1 does not describe a lipid emulsion for parenteral nutrition containing GPC, nor does it disclose a relevant concentration of GPC in such a lipid emulsion for parenteral nutrition.

[0016] US 20150132280 A1 describes dietary supplements for improving mood, energy, concentration, etc., including alpha-GPC among an extensive list of optional additional compounds, and optional other compounds that modulate the effects of theacrine. However, lipid emulsions for parenteral nutrition containing GPC are not mentioned.

[0017] In light of the present disclosure and without limiting the scope of the invention in any way, and in a first aspect of the present disclosure which may be combined with any other aspects listed herein unless otherwise specified, a lipid preparation for parenteral administration to a patient requiring choline supplementation comprises glycerophosphocholine (GPC) at a concentration of 0.1 g to 15.0 g per liter of lipid emulsion. The lipid preparation has an aqueous phase and an oil phase, comprises about 5% by weight to about 35% by weight of the oil phase based on the total weight of the lipid preparation, and exists in the form of an oil-in-water emulsion.

[0018] According to one aspect, the lipid emulsion contains GPC at a concentration of 2.0 to 12.0 g per liter of lipid emulsion. According to another aspect, the lipid emulsion contains GPC at a concentration of 4.0 to 11.0 g per liter of lipid emulsion. A lipid emulsion having a higher concentration of GPC as previously described is particularly useful for the treatment of pediatric patients.

[0019] According to another aspect, the lipid emulsion contains GPC at a concentration of 0.5 to 6.0 g per liter of lipid emulsion. According to another aspect, the lipid emulsion contains GPC at a concentration of 0.5 to 5.0 g per liter of lipid emulsion. Also, according to yet another aspect, the lipid emulsion contains GPC at a concentration of 0.5 to 3.0 g per liter of lipid emulsion. Lipid emulsions having lower concentrations of GPC as previously described are particularly useful for the treatment of adult patients.

[0020] According to another aspect of the present invention, the lipid emulsion additionally contains docosahexaenoic acid (DHA) at a concentration of 0.1 g to 15.0 g per 100 g of the oil phase. According to one aspect, the lipid emulsion contains DHA at a concentration of 0.25 g to 3.0 g per 100 g of the oil phase. According to another aspect, the lipid emulsion contains DHA at a concentration of 1.5 g to 2.5 g per 100 g of the oil phase. Also, according to yet another aspect, DHA exists in the form of triglycerides or ethyl esters, preferably in the form of triglycerides.

[0021] According to another aspect of the present invention, the lipid emulsion further contains arachidonic acid (ARA) at a concentration of 0.1 g to 15.0 g per 100 g of the oil phase. According to one aspect, ARA is present at a concentration of 1.5 g to 7.5 g per 100 g of the oil phase.

[0022] According to another aspect of the present invention, the lipid emulsion essentially does not contain eicosapentaenoic acid (EPA).

[0023] According to one aspect, the lipid emulsion contains EPA, wherein the ratio between DHA and EPA is 10:1 to 1000:1 (w / v), 10:1 to 200:1 (w / v), 10:1 to 100:1 (w / v), or 1:1 to 10:1 (w / v).

[0024] According to one aspect of the present invention, a lipid emulsion is provided in a single-chamber container for administration to a patient in need. The single-chamber container may be a glass vial as well as a flexible or rigid container.

[0025] According to one aspect of the present invention, the lipid emulsion according to the present invention is administered directly to a patient without mixing the lipid emulsion with other parenteral nutrition preparations, such as, for example, amino acid preparations or carbohydrate preparations.

[0026] In addition, according to another aspect, a lipid emulsion containing 0.5 g to 5.0 g GPC / L is used in the treatment of adult patients requiring choline supplementation. Specifically, the lipid emulsion is used in patients suffering from hepatic steatosis.

[0027] According to another aspect, the lipid emulsion for direct administration contains 2.0 g to 12.0 g GPC / L. Also, according to another aspect, a lipid emulsion containing 0.2 g to 0.9 g GPC / L is used for the treatment of adult patients requiring choline supplementation. Specifically, the lipid emulsion is used for patients suffering from hepatic steatosis.

[0028] According to another aspect, the lipid emulsion for direct administration is provided to a patient after adding one or more compounds, such as trace elements and / or vitamins, to the emulsion before or at the time of administration, wherein the concentration of glycerophosphocholine in the administered lipid emulsion is essentially unchanged.

[0029] According to another aspect of the present invention, a lipid emulsion is provided as one of several components in a multi-chamber bag (MCB) designed for parenteral administration of the reconstituted amount after mixing the formulations contained in each chamber. Such an MCB may have two, three, four, five, six or more chambers. The chambers of the MCB may have different sizes or the same size to accommodate various compositions and volumes. The chambers may be designed to contain volumes, for example, 1 to 5 ml, 5 to 10 ml, 10 to 50 ml, 50 to 100 ml, 100 to 250 ml, 250 ml to 500 ml, 500 to 1000 ml, or 1000 to 1500 ml. The MCB may be designed to have chambers located adjacent to each other. The chambers may have various shapes. The chambers may be oriented horizontally and / or vertically relative to each other. A specific small chamber may be designed to be located within another larger chamber, wherein, for example, the small chamber located within the other larger chamber may be accommodated and secured within the larger chamber by welding at least one edge of the small chamber between the welded seams of the surrounding larger chamber.

[0030] According to one aspect, a multi-chamber vessel for parenteral administration of its reconstituted content comprises a lipid emulsion according to the present invention in a first chamber and a carbohydrate preparation or amino acid preparation in a second chamber.

[0031] According to another aspect, a multi-chamber vessel for parenteral administration of its reconstituted content comprises a lipid emulsion according to the present invention in a first chamber, an amino acid preparation in a second chamber, and a carbohydrate preparation in a third chamber. Optionally, the multi-chamber vessel further comprises a fourth chamber containing vitamins and / or trace elements.

[0032] In addition, according to another aspect, the multi-chamber vessel comprises, in addition to the first, second, and third chambers, a vitamin preparation in a fourth chamber and a trace element preparation in a fifth chamber.

[0033] According to another aspect, a multi-chamber vessel includes a vulnerability barrier between chambers that can be opened before use to mix the components of all or selected chambers.

[0034] According to another aspect, the lipid emulsion provided in the multi-chamber bag contains GPC at a concentration of 0.1 g to 15.0 g / L.

[0035] In addition, according to another aspect, the lipid emulsion provided in the multi-chamber bag contains GPC at a concentration of 2.0 g to 12.0 g / L.

[0036] In addition, according to another aspect, the lipid emulsion provided within the multi-chamber bag contains GPC at a concentration of 4.0 g to 11.0 g / L. Such a multi-chamber bag is preferably used for the treatment of pediatric patients requiring parenteral nutrition.

[0037] According to yet another aspect of the present invention, a lipid emulsion provided in a multi-chamber bag contains GPC at a concentration of 0.5 g to 6.0 g / L. According to yet another aspect of the present invention, a lipid emulsion provided in a multi-chamber bag contains GPC at a concentration of 0.5 g to 5.0 g / L.

[0038] According to another aspect of the present invention, a lipid emulsion provided in a multi-chamber bag contains GPC at a concentration of 0.5 g to 3.0 g / L. Such a multi-chamber bag is preferably used for the treatment of adult patients requiring parenteral nutrition.

[0039] According to yet another aspect of the present invention, glycerophosphocholine is provided in a reconstituted solution from a multi-chamber bag, wherein GPC is present at a concentration of 0.01 g to 6.0 g per liter of the reconstituted solution. According to one aspect, the reconstituted solution contains glycerophosphocholine at a concentration of 0.01 g to 2.0 g / liter. According to yet another aspect, the reconstituted solution contains glycerophosphocholine at a concentration of 0.5 g to 5.0 g / liter.

[0040] According to another aspect of the present invention, glycerophosphocholine is provided in a parenteral nutrition composition reconstituted from a multi-chamber container comprising at least two, at least three, at least four, or at least five chambers, wherein at least one of the chambers is filled with an amino acid preparation, a carbohydrate preparation, or a lipid preparation, wherein glycerophosphocholine is present in the composition at a concentration of 0.01 g to 6.0 g / L. According to one embodiment, glycerophosphorylcholine is provided in a double-chamber bag or a three-chamber bag comprising at least one amino acid preparation and / or at least one carbohydrate preparation and / or at least one lipid preparation.

[0041] According to another aspect of the present invention, a method for providing choline to a patient is provided, wherein the patient requires parenteral nutrition and requires choline intake, and the method comprises administering a composition for parenteral administration characterized by containing glycerophosphocholine at a concentration of 0.01 g to 15.0 g per liter of composition.

[0042] According to another aspect of the present invention, a parenteral nutrition solution for the treatment of choline deficiency in pediatric and / or adult patients is provided, wherein the solution contains glycerophosphocholine at a concentration of 0.01 g to 15.0 g per liter of solution. According to one aspect, the parenteral nutrition solution is a lipid emulsion. According to another aspect, the lipid emulsion is a solution ready for direct administration to a patient. According to yet another aspect, the lipid emulsion is one of at least two, three, four, or five compositions provided within separate chambers of a multi-chamber bag.

[0043] According to another aspect of the present invention, a patient receiving glycerophosphocholine requires parenteral nutrition because oral and enteral nutrition is not possible, insufficient, or contraindicated. According to one aspect, the patient suffers from choline deficiency or is at risk of developing choline deficiency. According to an additional aspect, the patient suffers from hepatic steatosis or is at risk of developing hepatic steatosis. Also, according to another aspect, the patient suffers from parenteral nutrition-related liver disease, specifically hepatic steatosis.

[0044] According to another aspect of the present invention, the method of providing choline to a patient comprises administering glycerophosphocholine at a dose of 15 mg / kg / day to 300 mg / kg / day. According to one aspect, glycerophosphocholine is administered to pediatric patients at doses of 24 mg / kg / day to 210 mg / kg / day, 50 mg / kg / day to 150 mg / kg / day, and 70 mg / kg / day to 100 mg / kg / day.

[0045] In addition, according to another aspect of the present invention, the method of providing choline to a patient comprises administering glycerophosphocholine to an adult patient at doses of about 5 mg / kg / day to 50 mg / kg / day, 10 mg / kg / day to 30 mg / kg / day, and 20 mg / kg / day to 30 mg / kg / day.

[0046] According to another aspect of the present invention, a lipid emulsion or reconstituted solution containing glycerophosphocholine is administered by a peripheral or central catheter. Brief explanation of the drawing

[0047] It is understood that the drawings merely illustrate embodiments of the invention and should not be construed as limiting the scope of the disclosure, and that the disclosure is further described and explained in detail through the use of the accompanying drawings. Fig. 1 silver Example 1 This illustrates the development of alanine aminotransferase (ALT) function in the animal study described. The control group received only a saline solution (NaCl) or a lipid emulsion LE 10%, while the additional group received the same LE 10% lipid emulsion but contained three different choline sources: choline chloride, CDP-choline, and GPC in the lipid emulsion. The expression "high" indicates a concentration of 31 mmol / L of choline derivatives in the LE 10% lipid emulsion. The graph shows a decrease in ALT activity from day 5 to day 17, demonstrating an improvement in liver condition, which is most pronounced when GPC is present in the lipid emulsion. Fig. 2 Is Example 1In animal studies as described in (Step #3), the development of alanine aminotransferase activity in percentage units relative to normal values ​​in plasma is presented. The tested animals were exposed to either a saline solution or only an LE 10% lipid emulsion ("LE 10%"), where the lipid emulsion contained normal amounts of phytosterols ("Phyto") but no GPC. Additionally, another lipid emulsion containing LE 10% and a similar amount of phytosterols contained GPC at a concentration of 40 mmol / L ("LE 10% + GPC Step #2"). The third composition also contained GPC added at a concentration of 40 mmol / L and a reduced phytosterol content (phytosterol dosage is Fig. 2 It consisted of (provided in) and a lipid emulsion (LE 13%). Fig. 3a and Fig. 3b Is Example 1 Results of animal studies as described in [ ] and histopathological evaluation of the livers of animals subjected to parenteral nutrition with saline solution, LE 10% lipid emulsion, and LE 10% lipid emulsion supplemented with choline chloride, CDP-choline, and GPC, respectively ( Fig. 3a It represents ). Fig. 3a This suggests that the administration of a lipid emulsion containing GPC has the most significant effect on the development of vacuolation compared to all choline derivatives tested. Fig. 3b It also provides the results of administration of saline solution alone, lipid emulsion supplemented with DHA and ARA (LE 13%) (triangle), and lipid emulsion supplemented with DHA, ARA, and GPC (LE 13%) (square). Fig. 3b This suggests that the presence of DHA and ARA already has a significant positive effect on hepatic vacuolation. The expression "low" refers to a concentration of 15 mmol / L for each choline derivative. The expression "high" refers to a concentration of 31 mmol / L for the choline derivative. The grade is Example 3.3 It is defined as provided in. Fig. 4 Is Example 1The results of animal studies as described in [document] are presented, where the effects of parenteral administration of various lipid emulsions were determined based on the ratio of liver weight to body weight in percent. Study animals were treated with a saline solution, an LE 10% lipid emulsion without any choline derivatives ("lipid emulsion 10%"), and the same LE 10% lipid emulsion additionally containing 31 mmol / L of a choline derivative as presented. The expression "high" refers to the concentration of the choline derivative in the lipid emulsion of 31 mmol / L. Fig. 5 It presents exemplary photographs of grade 1 to 5 vacuolation determined from histopathological analysis of the livers of research animals ( Examples 1 and 3 (Reference). The presented tissue slides were stained with hematoxylin-eosin. Grades were assigned according to the following system: Fig. 5a : Grade 1 = Minimum / Very little / Very small fear, scattered. Fig. 5b Grade 2 = Slight / few / small lesions, lobular center to intermediate band. Fig. 5c : Grade 3 = Medium / medium number / medium size vacuoles, lobular center to medium band. Fig. 5d Grade 4 = Significant / Much / Large terror, centrilobular to periportal. Fig. 5e : Grade 5 = Severe / Much / Great Fear, Diffuse, All Liver Areas Invaded. Specific details for implementing the invention

[0048] The specific embodiments described herein generally relate to the field of parenteral nutrition. More particularly, some embodiments described herein relate to lipid emulsions for parenteral administration, wherein the lipid emulsion comprises an aqueous phase and an oil phase and exists in the form of an oil-in-water emulsion. Other embodiments described herein relate to a multi-chamber vessel for parenteral administration, wherein the vessel comprises the lipid emulsion in a first chamber and a carbohydrate preparation or amino acid preparation in a second chamber, optionally an amino acid in a third chamber where the second chamber contains a carbohydrate preparation, optionally a vitamin preparation or a trace element preparation in a fourth chamber, and optionally a trace element in a fifth chamber where the fourth chamber contains a vitamin preparation.

[0049] As used herein, the term “pediatrics” refers to newborns, including premature infants (preterm infants) of 1 month or less, full-term infants, and post-term infants; infants from 1 month to 1 year; children from 1 year to 12 years of age; and adolescents from 13 years to 21 years of age. As used herein, the term “adult” refers to a person 22 years of age or older.

[0050] As used herein, the term “essentially not containing” may refer to a composition containing 5% or less of a specified component, e.g., 3% or less, 2% or less, 1% or less, 0.5% or less, 0.1% or less, 0.05% or less, 0.02% or less, 0.01% or less, 0.005% or less, 0.002% or less, and / or 0.001% or less of a specified component. For example, a lipid emulsion or composition “essentially not containing EPA” may refer to a composition containing 5% or less of EPA, e.g., 3% or less, 2% or less, 1% or less, 0.5% or less, 0.1% or less, 0.05% or less, 0.02% or less, 0.01% or less, 0.005% or less, 0.002% or less, and / or 0.001% or less of EPA.

[0051] The terms “liver disease,” “liver injury,” and “liver damage” are used interchangeably herein. These terms refer to a condition of the liver defined by increased alanine aminotransferase (ALT) activity, histologically determined vacuolation, as well as an increase in the liver weight to body weight ratio. Specifically, these terms refer to hepatic steatosis that can be diagnosed by the said markers. More specifically, they refer to NAFLD. Even more specifically, they refer to NAFLD caused by choline deficiency, particularly choline deficiency induced by long-term total parenteral nutrition.

[0052] The present disclosure provides a lipid preparation for parenteral administration comprising glycerophosphocholine (GPC) as a source of choline for patients in need. The lipid preparation according to the present invention, intended for direct administration to patients requiring parenteral nutrition, contains GPC at a concentration of 0.1 g to 15.0 g per liter of lipid emulsion.

[0053] Pediatric patients generally require larger amounts of choline than adults. Therefore, lipid emulsions for direct administration to pediatric patients may contain GPC in the range of 2.0 g to 12.0 g per liter of lipid emulsion or 4.0 g to 11.0 g per liter of lipid emulsion. For example, lipid emulsions for direct administration to pediatric patients contain GPC in the range of 5.0 g to 12.0 g per liter of lipid emulsion, or 4.0 g to 10.0 g per liter of lipid emulsion.

[0054] Adults, on average, require less choline. Therefore, lipid emulsions for direct administration to adult patients may contain GPC in the range of 0.1 g to 6.0 g per liter of lipid emulsion. For example, lipid emulsions for direct administration to adult patients contain GPC in the range of 0.5 g to 5.0 g per liter of lipid emulsion, 0.5 g to 4.0 g per liter of lipid emulsion, or 0.5 g to 3.0 g per liter of lipid emulsion.

[0055] In the present invention, a lipid emulsion characterized by the presence of GPC in the lipid emulsion or reconstituted composition as described above, a multi-chamber vessel containing the same, and a composition reconstituted from the multi-chamber vessel are disclosed. In other cases, the composition of the lipid emulsion may vary over a wide range in specific components and their sources, as long as they can be safely used for intravenous administration to patients requiring parenteral nutrition.

[0056] A lipid emulsion containing GPC as disclosed above is used for the treatment of patients requiring parenteral nutrition. Patients who may benefit from the lipid emulsion according to the present invention may be adult patients and pediatric patients, wherein the GPC concentration may be adjusted as previously disclosed to address the higher choline needs of pediatric patients.

[0057] Patients who may benefit from the lipid emulsion according to the present invention, including pediatric and adult patients, are all patients who exhibit signs of choline deficiency, including signs of hepatic steatosis, or who are at risk of developing choline deficiency. Choline deficiency according to the present invention may be caused by total parenteral nutrition with parenteral nutrition products that do not provide the amount (equivalent) of choline required to sustain an adequate intake (AI) of, for example, about 550 mg / day (~8 mg / kg / day) for adult males, about 425 mg / day (~6 mg / kg / day) for adult females, and about 40 mg / day (~90 to 100 mg / kg / day) for pediatric patients.

[0058] The lipid emulsion according to the present invention is also suitable for avoiding or treating choline deficiency in newborns, including premature and extremely premature infants. As previously mentioned, choline plays a central role in the development of these pediatric patients, as it is an essential structural component of cell membranes, the neurotransmitter acetylcholine, and phospholipid synthesis, where it contributes to the synthesis of very low-density lipoproteins essential for the excretion of formal triglycerides from the liver. Given that choline is critical in the synthesis of acetylcholine, the most relevant neurotransmitter in cholinergic cells, a deficiency of choline may be linked to such memory deficits. Choline is linked to brain development, particularly in infants. Choline deficiency further activates cellular apoptosis, which can contribute to defect cell repair mechanisms. Importantly, choline deficiency leads to hepatic steatosis, which is a problem, particularly in infants, including premature infants, who require parenteral nutrition.

[0059] Accordingly, the present invention provides a lipid emulsion for the prevention and / or treatment of hepatic steatosis in patients, specifically in patients placed on total parenteral nutrition, wherein the lipid emulsion comprises glycerophosphocholine according to the present invention. However, the lipid emulsion of the present invention or a reconstituted composition containing it may be used for parenteral nutrition in patients who do not suffer from hepatic steatosis, because the presence of GPC can prevent the occurrence of such liver damage.

[0060] As previously mentioned, choline chloride and CDP-choline have already been used as choline derivatives for the supplementation of choline in patients. However, these compounds are not provided as components of readily available lipid emulsions for parenteral nutrition that do not require the addition of choline chloride or CDP-choline to the formulation when administered via a medical port for intravenous application or in a separate solution, thereby avoiding an additional step that carries the risk of mis-administration or contamination. In fact, it has been found that it is difficult to stabilize the aforementioned choline derivatives in parenteral nutrition solutions containing lipid emulsions ( Example 5 reference).

[0061] On the other hand, during the work leading to the present invention, it was found that GPC is not only more efficient in delivering choline to a patient in an effectively metabolizable form, but also can be further stabilized in a lipid emulsion and remains stable in a formulation reconstituted from a multi-chamber bag containing such a lipid emulsion in one of the chambers. Accordingly, the present invention provides a method for providing choline to a patient by parenteral nutrition as a component of an readily available lipid emulsion or a reconstituted formulation containing it, wherein the lipid emulsion comprises glycerophosphocholine according to the present invention.

[0062] Lipid preparations as disclosed herein are emulsions of an oil phase, a water phase, and an emulsifier that makes the two phases miscible. For lipid emulsions to be used as injectable emulsions for parenteral nutrition, the emulsion must be an oil-in-water (o / w) emulsion. This means that, as the emulsion must be miscible with blood, the oil is present in the internal (or dispersed) phase while the water is in the external (or continuous) phase. Accordingly, lipid emulsions as disclosed herein must also substantially not contain any suspended solids. Of course, lipid emulsions may contain additional components, including but not limited to antioxidants, pH modifiers, isotonic agents, vitamins, trace elements, and various combinations thereof. An overview of lipid emulsions, their composition, and use is provided, for example, in the literature [Driscoll, Journal of Parenteral and Enteral Nutrition 2017, 41, 125-134]. Further information on the use of lipid emulsions in parenteral nutrition for intensive care patients is provided, for example, in the literature [Calder et al., Intensive Care Medicine, 2010, 36(5), 735-749].

[0063] The oil phase of the lipid emulsion generally comprises polyunsaturated fatty acids, such as long-chain polyunsaturated fatty acids, which may exist as free acids, as ionized or salt forms of free acids, and / or as ester forms. Suitable esters of polyunsaturated fatty acids / long-chain polyunsaturated fatty acids include, but are not limited to, alkyl esters (e.g., methyl esters, ethyl esters, propyl esters, or combinations thereof) and triglyceride esters. In some cases, the long-chain polyunsaturated fatty acid has the structure R(C=O)OR' (where R is an alkenyl group having at least 17 carbon atoms, at least 19 carbon atoms, at least 21 carbon atoms, or at least 23 carbon atoms, and R' is absent, H, a counterion, an alkyl group (e.g., methyl, ethyl, or propyl), or a glyceryl group, and for example, R(C=O)OR' is a monoglyceride, diglyceride, or triglyceride). Polyunsaturated fatty acids for use in the lipid preparations disclosed herein include, but are not limited to, linoleic acid (LA), arachidonic acid (ARA), α-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), stearidonic acid (SDA), γ-linolenic acid (GLA), dihomo-γ-linolenic acid (DPA), and docosapentaenoic acid (DPA), in particular, DHA, ARA, and EPA (each of which may exist in free acid form, ionized or salt form, alkyl ester form, and / or triglyceride form). In some cases, polyunsaturated fatty acids and / or long-chain fatty acids are present in triglyceride form.

[0064] Typically, the lipid formulation comprises an oil phase of about 5% by weight to about 35% by weight based on the total weight of the lipid emulsion. For example, the oil phase of the lipid emulsion is present in amounts of about 8% by weight to 12% by weight, about 10% by weight to about 20% by weight, about 10% by weight to about 15% by weight, about 15% by weight to about 20% by weight, about 12% by weight to about 17% by weight, about 18% by weight to 22% by weight, and / or about 20% by weight based on the total weight of the lipid formulation. The oil phase typically, and preferably, contains omega-3 fatty acids in varying amounts depending on the source of the oil. The three types of omega-3 fatty acids involved in human metabolism are eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (both of which are typically found in marine fish oil) and α-linolenic acid (ALA) (typically found in vegetable oil).

[0065] The oil phase and its components may be derived from a single source or from different sources (see, for example, [Fell et al., Advances in Nutrition, 2015, 6(5), 600-610]). Among plant oils, currently used sources include, but are not limited to, soybean oil and olive oil, as well as coconut oil or palm kernel oil (medium-chain triglycerides (MCTs)). Another source is Cryptocodinium conii, which in some cases serves as a single source of the long-chain polyunsaturated fatty acid docosahexaenoic acid (DHA) ( Crypthecodinium cohnii ) and Shizochytrium( Schizochytrium It is algae including microalgae such as ) species. Marine oils used in parenteral lipid emulsions are processed from oily fish found mainly in cold waters, including, but not limited to, herring, sardines, and sardines. However, for example, krill, such as Antarctic krill (Eupaucia superbadana ( Euphausia superbaOther marine organisms, such as *Dana*, can be used as oil sources. Krill oil, for example, provides both EPA and DHA in amounts up to 35% w / w of fatty acids. As a component of lipid emulsions, krill oil is considered to possess anti-inflammatory properties due to the presence of DHA and EPA and is hypothesized to bind to endotoxins (Bonaterra et al: Krill oil-in-water emulsions protects against lipopolysaccharides-induced proinflammatory activation of macrophages in vitro. Marine Drugs (2017), 15:74).

[0066] Many microorganisms, including fungi, yeasts, and some bacteria, have the ability to synthesize significant amounts of LC-PUFAs, primarily ARAs (Sanaa et al., Journal of Advanced Research 11:3-13 (2018)). Related ARA-producers are, for example, the non-pathogenic fungus Mortierella spp., from which M. alpina ( M. alpina ) 1S-4The species and ATCC 32,222 produce up to 70% ARA of lipids. In some cases, the oil phase contains a blend of oils derived from the microalgae species *Schizochytrium* and the fungus *Mortierella alpina*. The oils in these blends include, but are not limited to, myristic acid (C14:0), palmitic acid (C16:0), palmitooleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2 n-6), gamma-linolenic acid (C18:3, n-6), alpha-linolenic acid (C18:3, n-3), arachidic acid (C20:0), ARA (C20:4, n-6), EPA (C20:5, n-3), behenic acid (C22:0), DPA (C22:5, n-3), DHA (C22:6, n-3), and lignoceric acid (C24:0). In some cases, the blends include ARA, DHA, and EPA. The content of various fatty acids, such as DHA, EPA, ARA, linoleic acid (LA), or alpha-linoleic acid (ALA), in oils from different sources is summarized, for example, in the literature [Fell et al., 2015].

[0067] In some embodiments of the present invention, the oil phase of the lipid emulsion essentially does not contain EPA and ARA, and EPA or ARA is added to the oil phase of the lipid emulsion alone or in combination. However, in some cases, where the oil phase already contains EPA and / or ARA, either of the fatty acids is added alone or in combination with each other to increase the amount of EPA and / or ARA in the lipid emulsion.

[0068] Accordingly, in some embodiments of the present invention, the oil phase of the lipid emulsion comprises ARA. In one embodiment, ARA is obtained from a fungus, preferably from Mortierella alpina. However, ARA may be obtained from various sources according to the present invention, and is not limited to ARA derived from a fungus. In some cases, ARA is present at a concentration of 0.1 g to 15 g per 100 g of oil phase, for example, 1.5 g to 7.5 g per 100 g of oil phase.

[0069] In some embodiments of the present invention, the oil phase contains DHA and EPA in varying ratios. In some cases, the oil phase contains more DHA than EPA, and in some cases, DHA is present but the oil phase essentially does not contain EPA. When both DHA and EPA are present, the ratio of DHA to EPA in the lipid emulsion is typically at least 10:1 (w / w), e.g., about 10:1 to about 1000:1 (w / w), about 20:1 to about 1000:1 (w / w), about 50:1 to about 1000:1 (w / w), about 100:1 to about 1000:1 (w / w), about 10:1 to about 200:1 (w / w), about 20:1 to about 150:1 (w / w), at least about 50:1 (w / w), at least about 100:1 (w / w), at least about 500:1 (w / w), and / or at least about 1000:1 (w / w).

[0070] In some cases, DHA is present at a concentration of 0.25 g to 3.0 g per 100 g of oil phase. In some cases, DHA is present in the oil phase of the lipid emulsion at a concentration of 1.5 g to 2.5 g per 100 g of oil phase. In some cases, the DHA is present in the form of a triglyceride or an ethyl ester, preferably in the form of a triglyceride.

[0071] An exemplary lipid emulsion according to the present disclosure contains glycerophosphocholine (GPC) at a concentration of 0.1 g to 2.0 g per liter of lipid emulsion. Such lipid emulsions may be used for direct parenteral administration. In certain embodiments, the lipid emulsion contains glycerophosphocholine (GPC) at a concentration of 0.1 g to 2.0 g per liter of lipid emulsion and additionally contains docosahexaenoic acid (DHA) at a concentration of 0.1 g to 5.0 g per 100 g of oil phase. Such lipid emulsions may essentially not contain EPA or may contain small amounts of EPA, wherein the ratio between DHA and EPA is 10:1 to 1000:1 (w / v) or 10:1 to 200:1.

[0072] In certain embodiments, the lipid emulsion contains glycerophosphocholine (GPC) at a concentration of 0.1 g to 2.0 g per liter of lipid emulsion, docosahexaenoic acid (DHA) at a concentration of 0.1 g to 5.0 g per 100 g of oil phase, and / or arachidonic acid (ARA) at a concentration of 0.1 g to 15 g per 100 g of oil phase. In certain embodiments, the lipid emulsion contains DHA and ARA in a DHA to ARA ratio of 10:1 to 1:5 (w / w) in the lipid emulsion. Such lipid emulsions may also essentially not contain EPA or may contain a small amount of EPA, wherein the ratio between DHA and EPA is 10:1 to 1000:1 (w / v) or 10:1 to 200:1.

[0073] The lipid emulsions disclosed herein contain reduced levels of phytosterols in some cases. In such cases, the oil used in the lipid emulsion is typically depleted of phytosterols by an amount of at least 25% of the total amount of phytosterols initially present in the oil, for example, by at least 40%, at least 50%, at least 60%, and / or at least 75%. In some cases, the lipid emulsion contains 70 mg or less of phytosterols per 100 g of oil phase, for example, about 15 mg to about 35 mg of phytosterols per 100 g of oil phase, and / or about 25 mg or less of phytosterols per 100 g of oil phase. The removal or depletion of phytosterols may be carried out by known processes, for example, short-path distillation, filtration after activated carbon treatment, supercritical CO2 chromatography, or chromatographic purification.

[0074] The lipid emulsions disclosed herein may further comprise additional components, such as surfactants (also referred to as emulsifiers), co-surfactants, isotonic agents, pH adjusters, and antioxidants. Generally, surfactants are added to stabilize the emulsion by reducing interfacial tension between the oil phase and the aqueous phase. Surfactants typically comprise a hydrophobic portion and a hydrophilic portion, and the amount of surfactant / emulsifier included in the formulation is determined based on the amount required to achieve the desired level of stabilization of the emulsion. Typically, the amount of surfactant in the lipid preparation is about 0.01 wt% to about 3 wt%, for example, about 0.01 wt% to about 2.5 wt%, about 0.01 wt% to about 2.3 wt%, about 0.02 wt% to about 2.2 wt%, about 0.02 wt% to about 2.1 wt%, about 0.02 wt% to about 2 wt%, about 0.05 wt% to about 1.8 wt%, about 0.1 wt% to about 1.6 wt%, about 0.5 wt% to about 1.5 wt%, about 0.8 wt% to about 1.4 wt%, about 0.9 wt% to about 1.3 wt%, about 1 wt% to about 1.2 wt%, and / or about 1.2 wt% based on the total weight of the lipid preparation. Suitable surfactants and co-surfactants include surfactants approved for parenteral use, which include but are not limited to phospholipids (e.g., egg phosphatidides and soy lecithin), oleate salts, and combinations thereof. Krill oil may also be used as an emulsifier in a lipid emulsion, wherein the lipid emulsion contains about 0.5 to 2.2 wt% krill oil based on the total weight of the emulsion, wherein the emulsion does not contain egg yolk lecithin (US 2018 / 0000732 A1). Another exemplary surfactant is lecithin, including both natural and synthetic lecithin, such as lecithin derived from eggs, corn or soybeans, or mixtures thereof.In some cases, lecithin is included in an amount of about 1.2% based on the total weight of the lipid preparation.

[0075] In some cases, lipid emulsion formulations contain a co-surfactant. Typically, the amount of co-surfactant in the lipid formulation is less than the amount of surfactant, and typically the amount of co-surfactant in the formulation is about 0.001 wt% to about 0.6 wt%, e.g., about 0.001 wt% to about 0.55 wt%, about 0.001 wt% to about 0.525 wt%, about 0.001 wt% to about 0.5 wt%, about 0.005 wt% to about 0.5 wt%, about 0.01 wt% to about 0.4 wt%, about 0.02 wt% to about 0.3 wt%, about 0.03 wt% to about 0.2 wt%, about 0.04 wt% to about 0.1 wt%, and / or about 0.05 wt% to about 0.08 wt% based on the total weight of the lipid formulation. An exemplary co-surfactant is an oleate, e.g., sodium oleate. In some cases, the lipid preparation contains lecithin and oleate as a surfactant and a co-surfactant, e.g., in amounts of 1.2% lecithin and 0.03% oleate. In some cases, sodium oleate is contained in an amount of about 0.03% by weight based on the total weight of the lipid preparation.

[0076] An isotonic agent may be added to a lipid emulsion to adjust the osmolarity of the lipid emulsion to a desired level, e.g., a physiologically acceptable level. Suitable isotonic agents include, but are not limited to, glycerol. Typically, the lipid emulsion formulation has an osmolarity of about 180 to about 300 milliosmolar / liter, e.g., about 190 to about 280 milliosmolar / liter, and / or about 200 to about 250 milliosmolar / liter. In some cases, the lipid emulsion contains the isotonic agent in an amount of about 1 wt% to about 10 wt%, e.g., about 1 wt% to about 5 wt%, about 1 wt% to about 4 wt%, and / or about 2 wt% to about 3 wt% based on the total weight of the lipid formulation. In some cases, the lipid emulsion formulation contains about 2 wt% to about 3 wt% of glycerol.

[0077] pH modifiers may be added to lipid emulsions to adjust the pH to a desired level, for example, a physiologically acceptable pH for parenteral use. Suitable pH modifiers include, but are not limited to, sodium hydroxide and hydrochloric acid. Typically, lipid emulsion formulations have a pH of about 6 to about 9, e.g., about 6.1 to about 8.9, about 6.2 to about 8.8, about 6.3 to about 8.7, about 6.4 to about 8.6, about 6.5 to about 8.5, about 6.6 to about 8.4, about 6.7 to about 8.3, about 6.8 to about 8.2, about 6.9 to about 8.1, about 7 to about 8, about 7.1 to about 7.9, about 7.2 to about 7.8, about 7.3 to about 7.7, about 7.4 to about 7.6, about 7, about 7.5, and / or about 8.

[0078] Lipid preparations may additionally contain antioxidants. Suitable antioxidants may be pharmaceutically acceptable antioxidants, which include, but are not limited to, tocopherols (e.g., gamma tocopherol, delta tocopherol, alpha tocopherol), ascorbyl palmitate, or combinations thereof. In some cases, lipid emulsion preparations contain antioxidants in amounts of about 0 to about 200 mg / L, e.g., about 10 to about 200 mg / L, about 40 to about 150 mg / L, about 50 to about 120 mg / L, or about 75 to about 100 mg / L of antioxidant(s), e.g., vitamin E.

[0079] The aqueous (or water) phase of all intravenous lipid emulsions must comply with pharmacopoeial requirements that make it suitable for injection, namely, the water must be sterile water for injection.

[0080] Additionally, the present disclosure includes a lipid emulsion in which GPC is added, for example, to a lipid emulsion or reconstituted solution that is immediately available through a medical port and then administered to a patient. GPC may be provided in the form of a suitable solution, for example, in a vial or in a suitable flexible or rigid container, or may be provided in a lyophilized form, for example, in a glass vial, wherein the lyophilized GPC is dissolved in a suitable solvent before being added to the lipid emulsion or reconstituted solution. GPC may be present as a single active ingredient in such solution or lyophilized product for addition to the lipid emulsion or reconstituted solution, or may be present in combination with at least one additional active ingredient, for example, a vitamin, a trace element, DHA, EPA and / or ARA. Accordingly, according to another embodiment of the present invention, a method of providing GPC to a patient is provided in which GPC is added to a lipid emulsion or a reconstituted solution from an MCB to provide a formulation for parenteral nutrition before administration.

[0081] As previously mentioned, a lipid emulsion may also be present in one of the chambers of a multi-chamber bag. Accordingly, the present disclosure also provides a multi-chamber container for parenteral administration of nutritional preparations. For example, the container may exist in the form of a bag having a plurality of compartments or chambers. The container, e.g., a bag, comprises at least two chambers, but may also contain three, four, or five chambers, and in one preferred embodiment, two or three chambers. Suitable containers, including soft bags, are typically sterile, non-pyrogenic, single-use, and / or ready for immediate use. Multi-chamber containers are particularly useful for maintaining pediatric and / or neonatal parenteral nutrition products and generally provide a carbohydrate preparation as disclosed herein in the first chamber, an amino acid preparation as disclosed herein in the second chamber, and a lipid preparation as disclosed herein in the third chamber of the container.

[0082] A multi-chamber vessel, such as a 3-chamber bag, may include vertical chambers. Suitable multi-chamber vessels are disclosed in U.S. Patent Publication No. 2007 / 0092579. For example, a multi-chamber vessel may be configured as a bag comprising two or three adjacent chambers or compartments. If desired, the chambers of the multi-chamber vessel are separated using a brittleness barrier or an openable seal (e.g., a release seal or a brittleness seal). The multi-chamber vessel may also include three chambers for accommodating lipid emulsions, carbohydrate preparations, and amino acid preparations, and may further include at least one, in certain embodiments, two or three smaller chambers containing, for example, vitamin preparations and / or trace element preparations. In one specific embodiment, the multi-chamber vessel of the present invention has a first chamber containing a lipid emulsion according to the present invention, a second chamber containing an amino acid preparation, a third chamber containing a carbohydrate preparation, a fourth chamber containing a vitamin preparation, and a fifth chamber containing a trace element preparation.

[0083] The openable seal of the multi-chamber container enables the storage of formulations within a single container that should not be stored as a mixture for an extended period, by allowing the formulations to be stored separately and mixed / reconfigured immediately before administration. Opening of the seal enables communication between chambers and mixing of the contents of each chamber. The outer seal of the multi-chamber container is a strong seal that does not open under supplied fluid pressure to open weaker release seals or brittle seals between chambers. In some embodiments, the openable seal of the multi-chamber container may be designed to enable mixing or reconfiguration of only selected chambers of the multi-chamber container, for example, mixing of a lipid emulsion with a vitamin chamber and an amino acid chamber if desired.

[0084] A multi-chamber vessel may be provided with instructions describing the desired order of opening the release seals so that the component fluids are mixed in the desired order. The opening of the seals in the desired order may be facilitated by varying the unsealing strength of two or more release seals. For example, the unsealing strength of the release seal to be opened first may be 1 / 3 to 1 / 2 of the unsealing strength required to open the release seal to be opened second.

[0085] When the lipid emulsion according to the present invention is contained in a multi-chamber bag, the concentration of GPC in the lipid emulsion may be adapted as necessary to reach the dose of GPC as previously disclosed. However, the lipid emulsion in the multi-chamber container may also contain GPC at a concentration as previously disclosed for the lipid emulsion.

[0086] As used herein, "reconstituted solution" refers to a solution for parenteral administration produced by mixing the contents of the chambers of a multi-chamber container before use.

[0087] A lipid emulsion contained within one chamber of a multi-chamber bag may contain GPC at a concentration of 0.1 g to 15.0 g / liter. In some embodiments, the concentration of GPC in such a multi-chamber vessel may be 1.0 g to 12 g per liter of lipid emulsion, 1.0 g to 10.0 g per liter of lipid emulsion, 2 g to 9.0 g per liter of lipid emulsion, 1.0 g to 5.0 g per liter of lipid emulsion, or 2.0 g to 4.0 g per liter of lipid emulsion.

[0088] As mentioned above, a typical component of a multi-chamber vessel for providing a formulation for parenteral nutrition is an amino acid and / or carbohydrate formulation. The amino acid formulation comprises a sterile aqueous solution of one or more amino acids and one or more electrolytes. Typically, the amino acid formulation comprises about 2 g to about 10 g of amino acids per 100 mL of amino acid formulation, for example, about 3 g to about 9 g and / or about 5 g to about 7 g of amino acids per 100 mL of amino acid formulation. Typical amino acids included in the amino acid formulation include, for example, isoleucine, leucine, valine, lysine, methionine, phenylalanine, threonine, tryptophan, arginine, histidine, alanine, aspartic acid, cysteine, glutamic acid, glycine, proline, serine, tyrosine, ornithine, and taurine. Additionally, the tyrosine content can be increased by adding, for example, glycyl-tyrosine dipeptide or acetyl-tyrosine (Ac-Tyr). However, typically, glycyl-tyrosine dipeptide has improved pharmacokinetics compared to Ac-Tyr, which is eliminated more rapidly by the kidneys, thereby attenuating the release of tyrosine into the blood.

[0089] Amino acid preparations may additionally contain electrolytes, such as sodium, potassium, calcium, magnesium, and / or phosphate ions. For example, the amino acid preparation contains, per 100 mL of amino acid preparation, about 0.1 mmol to about 10 mmol of sodium (e.g., about 3.75 mmol to about 10 mmol of sodium), about 0.1 mmol to about 10 mmol of potassium (e.g., about 3.75 mmol to about 6.90 mmol of potassium), about 0.05 mmol to about 1.0 mmol of magnesium (e.g., about 0.05 mmol to about 0.11 mmol and / or about 0.38 mmol to about 0.65 mmol of magnesium), about 0.1 mmol to about 10 mmol of calcium (e.g., about 1.13 mmol to about 5.10 mmol of calcium), about 0.1 mmol to about 10 mmol of phosphate (e.g., about 0.94 mmol to about 5.10 mmol of phosphate), and 10 mmol or less of chloride (e.g., 5.6 mmol or less of chloride). It may be included. When calcium and phosphorus are present together in the same heat-sterilized solution, insoluble calcium phosphate precipitation may occur. When using organic salts of phosphorus such as sodium glycerophosphate 5·H2O or calcium glycerophosphate, the amounts of calcium and phosphate can be increased without solubility issues and without providing an excess amount of sodium or chloride. In amino acid preparations, sodium may be provided in the form of sodium chloride, calcium may be provided in the form of calcium chloride 2·H2O or calcium gluconate, magnesium may be provided in the form of magnesium acetate 4·H2O or magnesium chloride, and potassium may be provided in the form of potassium acetate.

[0090] Carbohydrate preparations provide a supply of calories, typically in the form of glucose. In particular, carbohydrate preparations provide a sufficient amount of carbohydrates to avoid adverse effects, such as hyperglycemia, observed in patients receiving parenteral nutrition. Typically, carbohydrate preparations contain about 20 to 50 grams of glucose per 100 mL of carbohydrate preparation.

[0091] The lipid emulsion of the present invention can be prepared according to generally known methods (e.g., see reference [Hippalgaonkar et al., AAPS PharmSciTech 2010, 11(4), 1526-1540]). Generally, water-soluble and oil-soluble components are dissolved in the aqueous phase and oil phase, respectively. Thus, GPC is dissolved in the aqueous phase. An emulsifier, such as phosphatidide, can be dispersed in the oil or aqueous phase. Both phases are appropriately heated and stirred to disperse or dissolve the components. Subsequently, the lipid phase is added to the aqueous phase, generally under controlled temperature and stirring (using a high-shear mixer), to form a homogeneously dispersed coarse or pre-emulsion. A pre-emulsion having a droplet size of less than 20 μm generally produces a unimode and physically stable microemulsion. Next, the pre-emulsion is homogenized at optimized pressure, temperature, and number of cycles (using a microfluidizer or high-pressure homogenizer) to further reduce droplet size and form a microemulsion. Factors such as the type and concentration of the oil phase and surfactant, operating temperature, pressure, and number of cycles can affect the average droplet size during high-pressure homogenization and microfluidization. Throughout the service life of the emulsion, the average droplet size and PFAT5 (volume-weighted percentage of fat globules ≥5 μm) of the injectable microemulsion must be ≤500 nm and ≤0.05%, respectively. Next, the pH of the resulting microemulsion is adjusted to the desired value, and the emulsion is filtered through a 1–5 μm filter. Subsequently, the microemulsion is transferred into a suitable container. Plastic containers are oxygen-permeable and / or contain oil-soluble plasticizers, which are typically avoided. If possible, and especially if certain components of the excipients and lipid emulsions are oxygen-sensitive, the entire process (filtration / preparation of coarse and fine emulsions) is typically carried out under a nitrogen atmosphere. Sterilization of the lipid preparations can be achieved by final thermal sterilization or by aseptic filtration.Final sterilization generally provides a greater guarantee of the sterility of the final product. However, if the components of the emulsion are thermally unstable, sterile filtration may be used. Sterilization by filtration requires emulsion droplet sizes to be less than 200 nm. Alternatively, aseptic processing may be used. However, this process is relatively equipment and labor-intensive and requires additional process validation data and justification during regulatory submission.

[0092] Accordingly, the lipid emulsion according to the present invention can be prepared by a step comprising the following:

[0093] (a) a step of separately heating the oil phase and the aqueous phase to a temperature of about 70°C to about 80°C under stirring;

[0094] (b) a step of adding glycerophosphocholine to the aqueous phase;

[0095] (c) a step of preparing a pre-emulsion by transferring the oil phase to the aqueous phase under stirring;

[0096] (d) a step of homogenizing the pre-emulsion under pressure at a temperature of about 40°C to 60°C;

[0097] (e) a step of adjusting the required volume and concentration by adding water at will;

[0098] (f) optionally adjusting the pH to a range of about 7.8 to 8.8; and

[0099] (g) A step of optionally sterilizing the lipid emulsion.

[0100] Sterilization may be performed by methods known in the relevant art, such as by heat, for example. Typically, steps (a) through (d) will be performed in the presence of an inert gas, such as N2, to avoid any oxidation reaction. The pressure used for homogenizing the pre-emulsion will vary over a wide range. Generally, this will be in the range of 100 to 1300 bar, 200 to 1000 bar, 300 to 800 bar, or 400 to 1100 bar.

[0101] As previously discussed, the present invention relates to lipid emulsions and compositions containing the same for avoiding or treating hepatic steatosis. Hepatic steatosis is defined as intrahepatic fat of at least 5% of liver weight. Simple accumulation of triacylglycerols within the liver may be hepatoprotective; however, prolonged hepatic lipid storage can lead to hepatic metabolic dysfunction, inflammation, and advanced forms of non-alcoholic fatty liver disease (NAFLD). NAFLD encompasses a spectrum of diseases ranging from simple steatosis to non-alcoholic steatohepatitis (NASH), which can progress to cirrhosis and hepatocellular carcinoma. Histologically, NASH is defined by the presence of macrobulbar steatosis, lobular inflammation, and hepatocellular swelling (Kneeman, Secondary Causes of nonalcoholic fatty liver disease, Therap Adv Gastroenterol 2012; 5: 199-207). Long-term total parenteral nutrition (TPN) may be one of the many causes of NAFLD. TPN can lead to the depletion of carnitine, a compound essential for the transfer of free fatty acids from the hepatic cytoplasm into the mitochondria for beta-oxidation. As previously mentioned, cytosolar concentrations of choline also decrease, which promotes lipid storage within hepatocytes. This lipid storage is linked to the fact that the dominant histological findings are steatosis, intrahepatic cholestasis, and duct duplication. Therefore, the effects of choline supplementation during TPN can also be determined by verifying the histological features of the liver. Such histological analysis enables the assessment of the degree of steatosis by analyzing the extent of vacuolation. Example 3This provides the results of these histopathological investigations conducted in animal studies. It is suggested that liver damage caused by parenteral nutrition following a methionine and choline-deficient diet can be effectively resolved by the presence of glycerophosphocholine in the PN solution. This also provides evidence that GPC offers significantly superior effects compared to other choline derivatives, specifically choline chloride and CDP-choline. The additional presence of DHA and ARA appears to provide additional synergistic benefits.

[0102] The serum alanine aminotransferase (ALT) assay is the most common laboratory test for the detection of liver disease. Because enzyme concentrations within a population form a continuous distribution, a cut-off concentration distinguishing a healthy liver from a liver with disease is not clearly defined. However, the normal upper limit of serum alanine aminotransferase is set at an average of 40 IU / L and ranges from 30 to 50 IU / L (Kim et al. BMJ 2004, 328:983). The above upper limit is used as the normal serum alanine aminotransferase concentration (also referred to herein as "aminotransferase" or "ALT") in relation to the present invention. Kim et al. also note that the association between aminotransferase concentration and the lethality from liver disease is significant. Accordingly, serum aminotransferase concentrations were used in connection with the present invention to determine the efficacy of glycerophosphocholine in the treatment of liver damage that may be induced by parenteral nutrition, for example, compared to any lipid emulsion that does not contain choline, and also compared to other choline sources including choline chloride and CDP-choline. Figures 1 and 2 It demonstrates the significantly superior effect of GPC in reducing ALT concentration.

[0103] ALT activity is determined by modifications recommended by the International Federation of Clinical Chemistry (IFCC) (also Example 2(Reference). ALT catalyzes the reaction of alpha-ketoglutarate and L-alanine to form L-glutamate and pyruvate. Under the action of LDH, pyruvate is converted to lactate, and NADH is converted to NAD. The decrease in NADH absorbance measured at 340 nm (the secondary wavelength is 700 nm) is directly proportional to the serum activity of ALT. This is a kinetic response. Generally, serum is used for analysis. Serum is separated from cells within 2 hours of collection and stored at -70°C until assay. Monitoring of ALT activity over time in animal models is used herein as a simple and definitive method to determine the efficacy of GPC in the treatment of liver disease, specifically liver damage induced by a choline-deficient diet ( Example 1 and 2 , Fig. 1 and 2 ).

[0104] Plasma-free choline concentrations can be determined by high-pressure liquid chromatography (HPLC) and gas chromatography-mass spectrometry according to protocols known in the relevant art (Pomfret et al.: Measurement of choline and choline metabolite concentrations using high pressure liquid chromatography and gas chromatography-mass spectrometry. Anal Biochem. 1989; 180: 85-90).

[0105] Methods for depleting phytosterols from oils, specifically from plant-derived oils such as olive or soybean oil, are known. For example, phytosterols can be removed as described in the literature [Ostlund et al., Am J Clin Nutr 2002, 75, 1000-1004] or as described in US 6303803 B1.

[0106] Examples

[0107] Example 1: Animal model

[0108] Animal studies were conducted to identify and confirm the most effective choline derivatives for use in parenteral nutrition solutions, specifically lipid emulsions, to avoid or treat and improve liver damage, specifically hepatic steatosis. Additionally, the study was provided to identify any possible metabolic interactions between the choline derivative found to be most effective and polyunsaturated fatty acids, specifically DHA and ARA.

[0109] The effects of GPC as well as combinations of DHA, ARA, and GPC were investigated in diseased animal models, where male Sprague-Dolly rats with steatosis induced by a choline and methionine deficiency (MCD) diet (SSNIFF MCD pelletized choline / methionine deficiency diet (SSNIFF Spezialdiaeten GmbH, Sost, Germany, product number E15653-94)) were maintained on parenteral nutrition. Steatosis was confirmed in each group by histopathology ( Example 3 ).

[0110] Parenteral nutrition was initiated after 14 days under the MCD diet. The first measurement was performed 5 days after parenteral nutrition, and subsequent measurements were taken at 9 and 17 days. Daily lipid intake was 2.31 g / kg / d. To the rats Table I As described in [document], it was divided into several groups.

[0111] Two groups were provided as controls, wherein the first group was treated with a saline solution (0.90% w / v NaCl in water for injection). The second group received a lipid emulsion ("LE 10%") containing purified olive oil (80.0 g / L) and purified soybean oil (20.0 g / L) in water for injection at pH 6-8; LE 10% additionally contained egg lecithin (6.0 g / L), glycerol (11.25 g / L), and sodium oleate (0.15 g / L). LE 10% contained 100 g / L of lipid. In specific experiments, the lipid emulsion "LE" was used at different concentrations, e.g., "LE 13%" or "LE 20%", indicating that the concentration was correspondingly higher.

[0112] Various choline derivatives selected from the group consisting of choline chloride, glycerophosphocholine (GPC), and cytidine diphosphate-choline (CDP-choline) were each added to LE 10% at two concentrations (low / high) ( Table I (Refer to Step #2). "Low" refers to a lipid emulsion concentration of 15 mmol / L, and "high" refers to a lipid emulsion concentration of 31 mmol / L. Choline chloride (2-hydroxyethyl)trimethylammonium chloride), and pharmaceutical secondary standards were purchased from Sigma-Aldrich (Merck KGaA, Germany). CDP-choline (prepared by fermentation) was purchased from TCI Europe NV (Antwerp, Belgium). Glycerophosphocholine prepared from egg phosphatidide by saponification was supplied by Lipoid GmbH (Germany). DHA (from the algae, *Schizochytrium* species) and ARA (from the fungus *Mortierella alpina*) were purchased from BASF AG (Germany). Vitamin E was purchased from ADM (Illinois, USA).

[0113] A "LE" lipid emulsion supplemented with a choline derivative was prepared as follows. In the first step, the raw phase is heated. For the oil phase, soybean and olive oil and egg phosphatidide, as well as DHA and ARA oil and vitamin E (for the Step 3 experiment), are heated to 75°C in an inox beaker under N2 protection while continuously stirring with Ultra-Turax. The aqueous phase is prepared by a mixture of glycerol, sodium oleate, and Milli-Q water, and is continuously heated to approximately 75°C in an inox beaker under N2 protection. If necessary, the choline derivative is added under continuous stirring. In the second step, the oil phase is added to the aqueous phase using a peristaltic pump under continuous stirring to prepare a pre-emulsion, which is then transferred to a high-shear inline disperser (e.g., Dispax Reactor® DR). The receiving vessel is closed and flushed with nitrogen. In the third step, the pre-emulsion is homogenized by passing it several times through a high-pressure homogenizer under an N2 atmosphere at 600 bar and a temperature of about 50°C.

[0114] Next, the emulsion is adjusted to the required volume (2 L) with milli-Q water and cooled to a temperature of approximately 18 to 28°C, preferably 20 to 25°C. The pH is adjusted with 0.1N NaOH to a range of 7.8 to 8.8, preferably about 8.3 to 8.8. The emulsion is filtered through a 4.5 μm filter, transferred into a 100 mL bag, and then overpouched using an oxygen absorber / indicator for subsequent use. Subsequently, the lipid emulsion is autoclaved at 121°C to achieve an F0 of at least 15 minutes.

[0115] Table I: Study Groups and Experiment Setups: A standard "LE" lipid emulsion was used in the experiment, and a specific amount of choline source was added to the lipid emulsion. In other cases, this test setup, which focuses on the effects of GPC itself, is " Step #2 It is referred to as ".

[0116]

[0117] In additional tests, the LE lipid emulsion according to the above was used. However, in each experiment, the phytosterol content was reduced as indicated. Vitamin E was supplemented to the phytosterol-depleted LE lipid emulsion in the test at a concentration of 40 μg / ml relative to the lipid emulsion ( Table II (Reference). Additionally, DHA was contained in the lipid emulsion at a final concentration of about 2 to 3 g / L. ARA was contained in the lipid emulsion at a final concentration of about 5 to 6 g / L. The concentration of GPC here was slightly higher than in Step 2 (40 mmol / L), but the dose was maintained at 0.77 mmol / kg / d as presented in Table I. This test setup, which also considers the synergistic effects of DHA and ARA in the formulation together with GPC, was labeled herein as "Step 3".

[0118] Table II: Study Groups and Experiment Setups: Phytosterol-depleted "LE" lipid emulsions as described above were used in the experiment, where DHA and ARA were added as indicated as well as GPC. In other cases, this test setup is " Step #3 It is referred to as ".

[0119]

[0120] Example 2: Aminotransferase (ALT) Development

[0121] To evaluate the effect of GPC on the liver status of the tested animals as described in Example 1, aminotransferase (ALT) activity in the plasma of the tested animals was tested 5 and 17 days after parenteral nutrition in both Step 2 and Step 3 setups. As previously described, liver damage (cholera) can be determined by increased aminotransferase levels. ALT activity was determined using an ADVIA 1800 blood biochemistry analyzer / IFCC variant (Siemens) according to the standard protocol as previously described.

[0122] To summarize the results of the experiment, the development of ALT activity in the tested rats Fig. 1 This is presented in ). As can be seen here, an increase in ALT activity is expected over 17 days with parenteral nutrition when using LE 10%. In contrast, ALT activity does not increase in the study group where the choline source is present in the lipid emulsion. However, the effect is most noticeable in the presence of GPC, where a clear decrease in ALT activity can be determined. On the other hand, choline chloride and CDP-choline provide a stable situation where ALT activity does not significantly increase or decrease. Importantly, phosphatidylcholine contained in the LE lipid emulsion(s) appears to have no effect, and from this, it is confirmed that choline provided in the form of phosphorylcholine in the lipid emulsion cannot be utilized.

[0123] After identifying GPC as a surprisingly effective choline derivative for reducing ALT activity and consequent liver damage, another experiment focused on the synergy between GPC and PUFA DHA and ARA. A lipid emulsion as described in Example 1 was additionally Table III It was used as presented in.

[0124] Fig. 2This summarizes the results regarding ALT activity in relation to changes in activity from normal values. The test included one experiment with a composition in which the phytosterol content of the lipid emulsion was reduced according to the Step 3 setup, as shown in Figure 2. The results also demonstrate that both the saline solution and the lipid emulsion without a choline source and with normal phytosterol content ("LE") resulted in an increase in ALT activity, which is evidence of progressive liver deterioration. On the other hand, regardless of phytosterol content, all lipid emulsions with GPC (40 mmol / L) provided a significant decrease in ALT activity. The phytosterol dose Fig. 2 Phytosterols were contained in LE 13% lipid emulsions at concentrations ranging from 40 μg / mL to 120 μg / L. Importantly, the phosphatidylcholine contained in the lipid emulsion(s) used herein appears to have no effect, and from this, it is confirmed that the body cannot utilize choline provided in the form of phosphorylcholine in the lipid emulsion.

[0125] Example 3: Determination of Histopathological Grade

[0126] Example 3.1: Preparation of tissue slides

[0127] A portion of the liver was preserved in buffered formalin, and the injection site and sampled macroscopic lesions, where applicable, were trimmed according to RITA guidelines (Ruehl-Fehlert et al., Exp Toxic Pathol 2003; 55, 91-106), embedded in paraffin wax, thinned to a thickness of approximately 4 micrometers, and stained with hematoxylin-eosin. Frozen liver samples were cryopresected into approximately 5 micrometer slides and stained with Oil Red. Tissue processing was performed at Citoxlab (France).

[0128] Example 3.2: Microscopic Examination

[0129] Microscopic examinations were performed on two liver slides (hematoxylin eosin and Oil Red O) from all study animals, covering both the injection site and macroscopic lesions. Peer review was conducted on at least 30% of the animals from the high-dose group and an appropriate number of slides from identified target organs to confirm that the results recorded by the study pathologist were consistent and accurate. After the completion of the pathology peer review, all tissue slides were returned to Citox Lab France for archival purposes. During microscopic examination, representative photographs were obtained at the pathologist's discretion.

[0130] Example 3.3: Average histological vacuolation

[0131] Liver samples obtained after 8 and 15 days, respectively, were examined as described above and assigned to five grades of hepatocyte vacuolation. Vacuolation was generally diffuse and involved the entire area in the case of Grade 5. The periportal area was less involved, and the vacuolations were smaller in size for lower grades. Grades were assigned according to the following definitions:

[0132] Grade 1 = Minimum / Very little / Very small fear, scattered.

[0133] Grade 2 = Slight / few / small lesions, lobular center to intermediate band.

[0134] Grade 3 = Medium / medium number / medium size phore, lobular center to medium band.

[0135] Grade 4 = Significant / Much / Large terror, centrilobular to periportal.

[0136] Grade 5 = Severe / Much / Great Fear, Diffuse, All Liver Areas Invaded.

[0137] Exemplary photos for each grade Fig. 5 It is presented in [document]. The presented tissue slides were stained with hematoxylin-eosin (HE).

[0138] After 8 days, administration of saline solution causes 4 out of 4 animals to show signs of liver damage with grade 5 vacuolation ( Refer to Table IV Parenteral nutrition with LE 13% in the presence of DHA and ARA already produces a positive effect on liver histopathology. Of the 5 animals, 2 still exhibit grade 5 vacuolation. However, the livers of 3 out of 5 animals could be assigned grade 4. In the presence of GPC in addition to DHA and ARA, liver vacuolation is further improved. Of the 5 animals, only 1 exhibits signs of grade 5 liver damage, while another 2 rats show grade 4 symptoms, and 2 show grade 3 symptoms. Finally, when only GPC is added to the lipid emulsion, none of the 5 rats show grade 5 liver damage. 3 show grade 4, 1 shows grade 3, and 1 even exhibits grade 2 vacuolation.

[0139] Table IV: Microscopic changes in the liver after 8 days

[0140]

[0141] After 15 days of parenteral nutrition, the above effects became even more pronounced. While the liver condition of animals fed a saline solution remained in an important state as expected, the presence of GPC in the lipid emulsion, as well as the presence of DHA, ARA, and GPC together, provided a clear and significant improvement in the liver condition of the animals. After 15 days, it also became clear that the presence of DHA and ARA in specific concentration ranges in addition to GPC provided even synergistically improved results. Fig. 3bThe effects of DHA, ARA, and GPC on the development of liver vacuolation were summarized in the grades defined above. The graph shows a significant improvement in liver condition in the presence of all three, DHA, ARA, and GPC, where GPC appears to have an additional remarkable effect in the already beneficial presence of DHA and ARA in the lipid emulsion.

[0142] The same histopathological test was performed to compare the effects of choline derivatives alone, that is, in the absence of DHA and ARA. Fig. 3a The results of the histopathological examination of the livers of the animal study group were summarized. The effect of GPC was significantly superior at both high (31 mmol / L) and low (15 mmol / L) GPC concentrations. CDP-choline (low concentration, 15 mmol / L) also had an effect. The CDP-choline group with a high concentration (31 mmol / L) could not be graded. The groups receiving choline chloride (high and low concentrations) as part of the lipid emulsion were superior compared to those using only saline or lipid emulsions without any choline derivatives. However, liver damage could not be resolved to any adequate degree with the lipid emulsion supplemented with choline chloride.

[0143] Table V: Microscopic changes in the liver after 15 days

[0144]

[0145] Example 4: Liver development relative to body weight

[0146] The study group described in Example 1 was further evaluated for the development of liver weight relative to body weight. An increase in liver weight that is not proportional to body weight gain is typical of liver damage, specifically steatosis, as previously described. Fig. 4It also presents the results of the ratio of liver weight to body weight in percentage units for a group of animals receiving the same lipid emulsion containing only a saline solution, LE 10%, and a choline derivative added at the previously described concentration ("high": 31 mmol / L). As can be seen, administration of the lipid emulsion alone results in an increase in the ratio, which indicates an increase in liver weight due to liver damage (steatosis). Both the addition of choline chloride or CDP-choline lead to a decrease in the ratio and, consequently, an improvement in liver condition. However, the most pronounced decrease in the ratio is also achieved in GPC. Furthermore, the phosphatidylcholine contained in the lipid emulsion(s) appears to have no effect.

[0147] Example 5: Stability of lipid emulsions in the presence of choline derivatives

[0148] In addition to investigating the efficacy of choline derivatives in resolving liver damage, the stability of each of the lipid emulsions prepared as described in Example 1 was further evaluated after sterilization ( Table VI (Reference). It was found that the stability of lipid emulsions was improved in the presence of GPC compared to choline chloride or CDP-choline. In fact, choline chloride and CDP-choline were found to destabilize lipid emulsions during sterilization (heating). Lipid emulsions containing choline chloride or CDP-choline resulted in phase separation of the lipid emulsion during sterilization. In the presence of GPC, the lipid emulsion remained stable.

[0149] Table VI: Stability of lipid emulsions upon sterilization in the presence of different choline derivatives

[0150]

Claims

Claim 1 A lipid emulsion for parenteral administration comprising glycerophosphocholine (GPC) at a concentration of 0.1 g to 15.0 g / liter, wherein the lipid emulsion comprises an aqueous phase and an oil phase of 5% to 35% by weight based on the total weight of the lipid emulsion, wherein the lipid emulsion comprises docosahexaenoic acid (DHA) at a concentration of 0.1 g to 5.0 g per 100 g of oil phase and arachidonic acid (ARA) at a concentration of 0.1 g to 15.0 g per 100 g of oil phase for the treatment of choline deficiency and liver damage by parenteral administration, and wherein the lipid emulsion does not contain eicosapentaenoic acid (EPA). Claim 2 A lipid emulsion for parenteral administration according to claim 1, wherein GPC is contained at a concentration of 2.0 g to 12.0 g / L. Claim 3 A lipid emulsion for parenteral administration according to claim 1, wherein GPC is contained at a concentration of 0.5 g to 5.0 g / L. Claim 4 delete Claim 5 delete Claim 6 delete Claim 7 In paragraph 1, a lipid emulsion configured for the treatment of pediatric or adult patients. Claim 8 A multi-chamber vessel for providing a parenteral nutrition preparation, wherein the multi-chamber vessel comprises a first chamber containing a lipid emulsion according to claim 1, wherein GPC is present at a concentration of 0.1 g to 15.0 g / L. Claim 9 In paragraph 8, a multi-chamber container having a second chamber containing an amino acid preparation or a carbohydrate preparation. Claim 10 A multi-chamber container according to claim 9, wherein the second chamber contains an amino acid preparation and the third chamber contains a carbohydrate preparation. Claim 11 A multi-chamber vessel for parenteral administration according to claim 10, wherein the multi-chamber vessel has a fourth chamber containing a vitamin or trace element. Claim 12 A multi-chamber vessel for parenteral administration according to claim 11, wherein the fourth chamber contains a vitamin and the fifth chamber contains a trace element. Claim 13 A lipid emulsion provided in a multi-chamber vessel according to claim 8, wherein GPC is present at a concentration of 0.1 to 15.0 g / liter. Claim 14 Parenteral nutrition composition reconstituted from a multi-chamber vessel according to paragraph 8. Claim 15 A parenteral nutrition composition according to claim 14, wherein GPC is present at a concentration of 0.01 g to 6.0 g per liter of reconstituted solution. Claim 16 A parenteral nutrition composition reconstituted from a multi-chamber vessel comprising an amino acid preparation, a carbohydrate preparation, and a lipid preparation, wherein glycerophosphocholine is present in the lipid preparation of the composition at a concentration of 0.01 g to 6.0 g / L, and the lipid emulsion comprises docosahexaenoic acid (DHA) at a concentration of 0.1 g to 5.0 g per 100 g of oil phase and arachidonic acid (ARA) at a concentration of 0.1 g to 15.0 g per 100 g of oil phase for the treatment of choline deficiency and liver damage by parenteral administration, and the lipid emulsion does not contain eicosapentaenoic acid (EPA). Claim 17 In paragraph 14, a parenteral nutrition composition for the treatment of pediatric or adult patients. Claim 18 delete Claim 19 delete Claim 20 delete Claim 21 delete Claim 22 delete Claim 23 delete Claim 24 delete Claim 25 delete Claim 26 delete Claim 27 A method for preparing a lipid emulsion of claim 1, comprising the following steps: (a) separately heating an oil phase and an aqueous phase to a temperature of 70°C to 80°C under stirring; (b) adding glycerophosphocholine to the aqueous phase; (c) preparing a pre-emulsion by transferring the oil phase to the aqueous phase under stirring; (d) homogenizing the pre-emulsion under pressure at a temperature of 40°C to 60°C; (e) optionally adding water to adjust the required volume and concentration; (f) optionally adjusting the pH to a range of 7.8 to 8.8; and (g) optionally sterilizing the lipid emulsion. Claim 28 A method according to claim 27, wherein at least steps (a) through (d) are performed in the presence of an inert gas. Claim 29 In paragraph 27, a method of sterilizing a lipid emulsion by heat. Claim 30 A lipid emulsion for parenteral administration configured to reduce ALT activity or liver damage and containing glycerophosphocholine (GPC) at a concentration of 0.1 g to 15.0 g / liter, wherein the lipid emulsion comprises an aqueous phase and an oil phase of 5% to 35% by weight based on the total weight of the lipid emulsion, wherein the lipid emulsion comprises docosahexaenoic acid (DHA) at a concentration of 0.1 g to 5.0 g per 100 g of oil phase and arachidonic acid (ARA) at a concentration of 0.1 g to 15.0 g per 100 g of oil phase for the treatment of choline deficiency and liver damage by parenteral administration, and wherein the lipid emulsion does not contain eicosapentaenoic acid (EPA). Claim 31 A parenteral nutrition composition reconstituted from a multi-chamber vessel, wherein the parenteral nutrition composition is configured to reduce ALT activity or liver damage and comprises an amino acid preparation, a carbohydrate preparation, and a lipid preparation, wherein glycerophosphocholine is present in the lipid preparation of the composition at a concentration of 0.01 g to 6.0 g / L, and wherein the lipid emulsion comprises docosahexaenoic acid (DHA) at a concentration of 0.1 g to 5.0 g per 100 g of oil phase and arachidonic acid (ARA) at a concentration of 0.1 g to 15.0 g per 100 g of oil phase for the treatment of choline deficiency and liver damage by parenteral administration, and wherein the lipid emulsion does not contain eicosapentaenoic acid (EPA). Claim 32 In paragraph 28, the method wherein the inert gas comprises N2.

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