Chemical synthesis of bioactive precursors to plasmalogens

Abstract

This disclosure relates to synthetic methods of making bioactive in vivo precursors to plasmalogen compounds and salts thereof and to making plasmalogens and their derivatives and salts thereof. This disclosure also relates to chemical precursors to bioactive compounds and salts thereof. This disclosure further relates to compositions comprising one or more compounds of this disclosure and the use of such compositions in preparing compounds useful in methods of treating diseases and conditions that are beneficially treated by administering plasmalogens or their derivatives or precursors that are transformed in vivo to plasmalogens.

Description

Ref: P329WO CHEMICAL SYNTHESIS OF BIOACTIVE PRECURSORS TO PLASMALOGENS TECHNICAL FIELD

[0001] This disclosure provides synthetic methods of making bioactive in vivo precursors to plasmalogen compounds and salts thereof and to making plasmalogens and their derivatives and salts thereof. This disclosure also relates to chemical precursors to the bioactive compounds and salts thereof. This disclosure also provides compositions comprising one or more compounds of this disclosure and the use of such compositions in preparing compounds useful in methods of treating diseases and conditions that are beneficially treated by administering plasmalogens or their derivatives or precursors that are transformed in vivo to plasmalogens. BACKGROUND

[0002] Plasmalogens comprise a group of plasmanyl- and / or plasmenyl- phospholipids. Plasmalogens are primarily present as alkenylphosphatidylcholine (PC) and alkenylphosphatidylethanolamine (PE) species, also known as PC(P) and PE(P), respectively. They are characterized by a cis-vinyl ether bond linking an alkyl chain to the sn-1 carbon position of the glycerol backbone. They also optionally have an acyl linked fatty acid ester bond in the sn-2 carbon position. Plasmalogens are often esterified with polyunsaturated fatty acids such as arachidonic acid (20:4) and the omega-3 fatty acid docosahexaenoic acid (22:6, a major constituent of fish oil), whereas the vinyl ether linked residue is usually otherwise saturated (i.e., no double bonds are present in the chain other than the vinyl ether group) or otherwise monounsaturated (i.e., one double bond is present in the chain in addition to the vinyl ether group.

[0003] Plasmalogens are essential membrane constituents that modulate membrane properties. LPX(O)s such as LPE(O) and LPC(O) are alkyl ether lipids and alkenyl ether lipids that are lyso alkyl or alkenyl containing phospholipids such as lysoalkylphosphatidylcholine or lysoalkylphosphatidylethanolamine. These are sub- classes of the broad family of naturally occurring ether lipids. Beside these two classes of compounds, there exists a multitude of other ether lipids that feature partly or completely substituted glycerol. Some of these compounds are more bioavailable than others in their ability to be incorporated into plasmalogens in vivo when fed orally and / or are more stable in mammalian digestive tract. 1Ref: P329WO

[0004] Ether lipid compounds may have a structure defined by Formula (I):I).

[0005] In compounds of Formula (I), the sn-1 carbon is labelled as above. R3includes phosphate and substituted phosphate. R1and R2can each be independently hydrogen, alkyl, alkenyl, acyl, or aryl, or substituted versions of these moieties as described herein. As used herein, the structural positions containing R1, R2, and R3are also referred to as positions P1, P2, and P3, respectively.

[0006] In Scheme I, some typical plasmalogens are shown in the top two structures, the boxes highlighting the sn-1 substitution as derived from Formula (I).Scheme I. Typical Plasmalogens.

[0007] Dietary plasmalogen modulation has 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, which is incorporated by reference herein, in its entirety. Extraction of lipids from krill oil results in a highly complex mixture of ether lipids. It is beneficial to produce ether lipids by a defined synthetic route to be able to accurately define the final resulting compound. It is also beneficial to theRef: P329WO production of compositions with known percentages of ether lipid precursors and ether lipid compounds in large amounts and with high purity.

[0008] Methods of using certain ether lipids and / or plasmalogen compositions are described in WO 2021 / 007623 A1 to Baker Heart and Diabetes Institute, having inventor Meikle, which is incorporated by reference herein in its entirety. Methods of making LPX(O)s are also disclosed in PCT / US2024 / 010129, entitled “FORMULATION AND USE OF PRECURSOR COMPOUNDS TO MODULATE PLASMALOGENS” to Baker Heart and Diabetes Institute and Pelagic AI Limited and having inventors Meikle, et al., which is incorporated herein by reference in its entirety. There are numerous approaches to the synthesis of these ether lipid compounds using either solely chemical synthesis or a combination of chemical and enzymatic synthesis strategies. Synthesis of the lysoalkenylphosphatidylethanolamine (LPE(P)) from commercially available 2,3-O-isopropylidene-sn-glycerol has been 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 numerous alkyl ether lipids (Gomes MAGB, Bauduin A, Le Roux C, Fouinneteau R, Berthe W, Berchel M, Couthon H, Jaffrès PA. “Synthesis of ether lipids: natural compounds and analogues,” Beilstein J. Org. Chem.2023 Sep 8;19:1299-1369). FIG.2 of Gomes loc cit. discusses a synthetic route that would be adaptable to production of the alkyl LPC(O) compounds of interest, providing the R-groups for the sn-1 fatty acid of interest and stopping at the lyso PAF step. FIG.6 of Gomes and further references disclosed therein describe how to produce the starting compound 1-O-alkylglycerol (i.e., compound 2.1 of FIG.2 of Gomes). Additionally, U.S. Pat. No.10,900,063, which is incorporated by reference herein in its entirety, describes the use of lipases to produce LPC(O)s.

[0009] Thus, there are numerous approaches to the synthesis of ether lipid compounds using either solely chemical synthesis or a combination of chemical and enzymatic synthesis strategies. However, the previously described synthetic and biosynthetic methods are complicated and expensive. Accordingly, it is desirable to 3Ref: P329WO provide a plasmalogen synthetic method which is convenient and economical and which provides improved yields. SUMMARY

[0010] Disclosed herein are convenient and economical synthetic methods of making bioactive precursors to plasmalogen compounds such as LPX(O)s and salts thereof as well as plasmalogen compounds and derivatives and salts thereof. These synthetic routes include methods of synthesis for plasmalogens and precursors of plasmalogens. These LPX(O)s prepared using intermediates of the disclosure may be used in the treatment of diseases, as nutraceuticals, for health maintenance, or as part of a normal diet as a food item.

[0011] In some embodiments, the disclosure provides a method for synthesizing Compound (6) or a salt thereof .

[0012] Withouteory, embodiments of methods of the disclosure provide certain advantages, including, but not limited to, the use of inexpensive starting materials and / or reagents, atom economical chemical transformations, and opportunities to use intermediate compounds in subsequent chemical transformations without purification. Moreover, as described herein, compounds employed in methods of the disclosure, including, but not limited to, Compound (6), are versatile intermediates that can be used in various embodiments. Additionally, without wishing to be bound by theory, it is understood that embodiments of methods of the disclosure are applicable to both syntheses of enantiomerically pure compounds and syntheses of racemic mixtures.

[0013] In some embodiments, the method comprises (i) reacting (R)-(2,2- dimethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1-OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2) 4Ref: P329WO .

[0014] In someembodiments, the method further comprises (ii) hydrolyzing Compound (2) in a mixture comprising acid to form Compound (3) .

[0015] In some ed further comprises (iii) reacting Compound (3) with a protecting group reagent in solution, the protecting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4) , wherein PG is a protecting g

[0016] In some embodiments, the method further comprises (iv) reacting Compound (4) in a mixture comprising an allylating reagent to form Compound (5).

[0017] In some embodiments, the method further comprises (v) deprotecting Compound (5) in solution to form Compound (6).

[0018] In some embodiments, the disclosure provides methods for synthesizing bioactive in vivo precursors to plasmalogen compounds. In some 5Ref: P329WO embodiments, the disclosure provides methods for synthesizing LPE(O)s. In some embodiments, the disclosure provides a method for synthesizing Compound (10) (depicted below in a non-ionic form) or a salt thereof .

[0019] Compound (10) and other structures disclosed herein may also be present in a zwitterionic form as pictured below for Compound (10).

[0020] mpound (10) in the non-ionic form is intended to include a depiction of, for example, Compound (10) in the zwitterionic form and vice versa.

[0021] In some embodiments, the method comprises (i) reacting (R)-(2,2- dimethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1-OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2) .6Ref: P329WO

[0022] In some embodiments, the method further comprises (ii) hydrolyzing Compound (2) in a mixture comprising acid to form Compound (3) .

[0023] In some ed further comprises (iii) reacting Compound (3) with a protecting group reagent in solution, the protecting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4) .

[0024] In somefurther comprises (iv) reacting Compound (4) in a mixture comprising an allylating reagent to form Compound (5).

[0025] In some embodiments, the method further comprises (v) deprotecting Compound (5) in solution to form Compound (6) .

[0026] In some ed further comprises (vi) phosphorylating the primary alcohol of Compound (6) to form Compound (7).

[0027] In some embodiments, the method further comprises (vii) reacting Compound (7) with ethanolamine in solution to form Compound (8) 7Ref: P329WO .

[0028] In soer comprises (viii) reacting Compound (8) in a mixture comprising acid to form Compound (9) (depicted below in a non-ionic form) .

[0029] ionic form as pictured below.

[0030] As used herein, a depiction of Compound (9) in the non-ionic form is intended to include a depiction of Compound (9) in the zwitterionic form and vice versa.

[0031] In some embodiments, the method further comprises (ix) deprotecting Compound (9) to form Compound (10) .8Ref: P329WO

[0032] In some embodiments, the method comprises reacting bis(2- cyanoethyl)-N,N-diisopropylphosphoramidite with Fmoc-protected aminoethanol to give the corresponding diisopropylphosphoramidite Compound (16) 6) and reacting Compound (1ompound (13).

[0033] In some embodiments, the method further comprises (vii-c) reacting Compound (13) with an oxidizing reagent to form Compound (14).

[0034] In some embodiments, the method further comprises (viii-c) selectively deprotecting Compound (14) at the secondary alcohol to form Compound (15) .9Ref: P329WO

[0035] In some embodiments, the method further comprises (ix-c) deprotecting Compound (15) to form Compound (10) .

[0036] ed from d

[0037] s for synthesizing bioactive in vivo precursors to plasmalogen compounds. In some embodiments, the disclosure provides methods for synthesizing LPC(O)s. In some embodiments, the disclosure provides a method for synthesizing Compound (12) or a salt thereof 10Ref: P329WO .

[0038] vi-a) reacting Compound (6) with 2-bromoethyl phosphorodichloridate to form Compound (9a).

[0039] In some embodiments, the method further comprises (vii-a) reacting Compound (9a) in a solution comprising a metal oxide and trimethylamine to form Compound (11) .

[0040] prises (viii-a) deprotecting Compound (11) to form Compound (12) .

[0041] vi-b) phosphorylating the primary alcohol of Compound (6) to form Compound (7) 11Ref: P329WO.

[0042] In some embodiments, the method further comprises (vii-b) reacting Compound (7) with ethanolamine in solution to form Compound (8) .

[0043] In ser comprises (viii-b) reacting Compound (8) in a mixture comprising an acid to form Compound (9).

[0044] In some embodiments, the method further comprises (ix-b) methylating compound (9) to form Compound (11). In some embodiments, methylating Compound (9) comprises reacting Compound (9) with methyl bromide, methyl iodide, or dimethyl sulfate.

[0045] In some embodiments, the method further comprises (x-b) deprotecting Compound 11 to form Compound (12) 12Ref: P329WO . In some embodiments, (x-b)ne catalyst and under an inert atmosphere. In some embodiments, the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. In some embodiments, a catalyst derived from any of the foregoing or generated in situ from any of the foregoing by combining any of the foregoing with a ligand, such as a phosphine ligand, such as triphenylphosphine, tri-tert-butylphosphine, or 1,1′- ferrocenediyl-bis(diphenylphosphine).

[0046] In some embodiments, the method comprises (1-a) reacting R1-OH with (2R)-(-)-glycidyl tosylate wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group (Compound (18)) to form Compound (19) ).

[0047] In some efurther comprises (1-b) reacting Compound (19) with a benzylating reagent in solution, the benzylating reagent reacting with the secondary alcohol of Compound (19) to form Compound (20) ), wherein Ar is selected from phyl.

[0048] In some embodiments, the method further comprises (1-c) reacting Compound (20) with a metal acetate to form Compound (21) 13Ref: P329WO ).

[0049] In someurther comprises (1-d) hydrolysing Compound (21) to form Compound (22) ).

[0050] In some ed further comprises (1-e) reacting Compound (22) with ethylene chlorophosphate to form Compound (23) ).

[0051] In somerther comprises (1-f) reacting Compound (23) in a solution comprising trimethylamine to form Compound (24) ).

[0052] Incomprises (1-g) deprotecting Compound (24) to form Compound (12) .

[0053] ted from 14Ref: P329WO d

[0054] In some embodiments, the disclosure provides methods for synthesizing bioactive in vivo precursors to plasmalogen compounds. In some embodiments, the disclosure provides a method for synthesizing Compound (25) or a salt thereof .

[0055] (2-a) providing Compound (26), wherein each of R1and R3is independently selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, .15Ref: P329WO

[0056] In some embodiments, the method further comprises (2-b) phosphorylating the primary alcohol of Compound (26) to form Compound (27) .

[0057] In soher comprises (2-c) reacting Compound (27) with ethanolamine in solution to form Compound (28) .

[0058] In ser comprises (2-d) reacting Compound (28) in a mixture comprising acid to form Compound (25) .

[0059] rs to plasmalogen compounds and salts thereof. Further disclosed herein are compositions comprising one or more bioactive in vivo precursors to plasmalogen compounds or salts thereof. In some embodiments, the disclosure provides a compound selected from 16Ref: P329WOCompo ,Compound ,17Ref: P329WO , , ,p ,and a salt of any of the foregoing, wherein each of R1and R3is independently selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, such as wherein each of R1and R3is independently selected from a C16-C18 alkyl group, a C16- C18 alkenyl group, and a C16-C18 acyl group. In some embodiments, the disclosure provides a composition comprising the same. EMBODIMENTS E-1. A method for synthesizing Compound (6) 18Ref: P329WO , or a salt thereof, the method comprising:ethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1- OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2); );cting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4), wherein PG is a protecting group;) in a mixture comprising an allylating reagent to form Compound (5); andd (5) in solution to form Compound (6) 19Ref: P329WO. E-2. A method for synthesizing Compound (10) , or a salt thereof, the method(i) reacting (R)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1- OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2); );cting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4), wherein PG is a protecting group;20Ref: P329WO (iv) reacting Compound (4) in a mixture comprising an allylating reagent to form Compound (5);(v) deprotecting Compound (5) in solution to form Compound (6);primary alcohol of Compound (6) to form Compound (7);thanolamine in solution to form Compound (8);ixture comprising acid to form Compound (9);and (ix) deprotecting Compound (9) to form Compound (10) 21Ref: P329WO .ein compound (6) is further reacted to form Compound (12) , or a salt thereof, the method(vi-a) reacting Compound (6) with 2-bromoethyl phosphorodichloridate to form Compound (9a);prising a metal oxide and trimethylamine to form Compound (11);and (viii-a) deprotecting Compound (11) to form Compound (12) 22Ref: P329WO .ein Compound (6) is further reacted to form Compound (12) , or a salt thereof, the method(vi-b) phosphorylating the primary alcohol of Compound (6) to form Compound (7);(vii-b) reacting Compound (7) with ethanolamine in solution to form Compound (8);mixture comprising an acid to form Compound (9);(ix-b) methylating compound (9) to form Compound (11); 23Ref: P329WO andnd (12) .ein compound (6) is further reacted to form Compound (10) , or a salt thereof, the method(vi-c) reacting bis(2-cyanoethyl)-N,N-diisopropylphosphoramidite with Fmoc-protected aminoethanol to give the corresponding diisopropylphosphoramidite Compound (16) (16)g p (16) with Compound (6) to form Compound (13); 24Ref: P329WO(vii-c) reacting Compound (13) with an oxidizing reagent to form Compound (14)(viii-c) selectively deprotecting Compound (14) at the secondary alcohol to form Compound (15); andd (10) .ents 1 to 5, wherein (i) comprises reacting Compound (1) and the R1-OSO2R2or the R1-X in the presence of a base. E-7. The method of any one of embodiments 1 to 6, wherein the base used in (i) is an alkali metal hydroxide, an alkali metal alkoxide, or alkali metal hydride, 25Ref: P329WO such as sodium hydroxide, potassium hydroxide, sodium tert-butoxide, potassium tert-butoxide, sodium hydride, or potassium hydride. E-8. The method of any one of embodiments 1 to 7, wherein (i) comprises heating the solution. E-9. The method of any one of embodiments 1 to 8, wherein (i) comprises heating the solution at a temperature range of about 60°C to 100°C. E-10. The method of embodiment 9, wherein (i) comprises heating the solution at a temperature of about 80°C. E-11. The method of any one of embodiments 1 to 7, wherein (i) comprises reacting Compound (1) with the R1-OSO2R2or R1-X at room temperature. E-12. The method of embodiment 1, wherein (i) comprises reacting Compound (1) with R1- OSO2R2. E-13. The method of any one of embodiments 1 to 12, wherein (i) comprises adding tetrabutylammonium bromide to the solution. E-14. The method of embodiment 13, wherein the addition of tetrabutylammonium bromide occurs after the addition of R1-OSO2R2or R1-X. E-15. The method of any one of embodiments 1 to 14, wherein (i) further comprises purifying Compound (2) before hydrolyzing Compound (2). E-16. The method of any one of embodiments 1 to 14, wherein, in (i), Compound (2) is not purified before Compound (2) is hydrolyzed. E-17. The method of any one of embodiments 1 to 16, wherein (ii) comprises dissolving Compound (2) in a solvent before hydrolyzing Compound (2). E-18. The method of embodiment 17, wherein the solvent is tetrahydrofuran. E-19. The method of any one of embodiments 1 to 18, wherein the acid in (ii) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate. E-20. The method of any one of embodiments 1 to 19, wherein (ii) comprises heating the mixture to 50°C to 80°C. E-21. The method of embodiment 20, wherein (ii) comprises heating the mixture to 65°C. 26Ref: P329WO E-22. The method of any one of embodiments 1 to 21, wherein (ii) comprises extracting Compound (3) from the mixture with 2-methyltetrahydrofuran. E-23. The method of any one of embodiments 1 to 22, wherein, in (iii), the solution comprises one or more of pyridine, dichloromethane, imidazole, and dimethylformamide. E-24. The method of any one of embodiments 1 to 23, wherein, in (iii), the temperature of the solution is maintained at from about 0°C to room temperature. E-25. The method of any one of embodiments 1 to 24, wherein the protecting group reagent in (iii) is selected from tert-butyldimethylsilyl chl / oride, trimethylsilyl chloride, triethylsilyl chloride, triisopropylsilyl chloride, a benzyl halide, and a triphenylmethyl halide. E-26. The method of embodiment 25, wherein the protecting group reagent in (iii) is tert-butyldimethylsilyl chloride. E-27. The method of any one of embodiments 1 to 26, wherein the allylating reagent in (iv) is an allyl halide. E-28. The method of embodiment 27, wherein the allyl halide is allyl bromide. E-29. The method of any one of embodiments 1 to 28, wherein, in (iv), the mixture comprises dry tetrahydrofuran. E-30. The method of any one of embodiments 1 to 29, wherein, in (iv), the mixture comprises a metal hydride and / or a metal alkoxide. E-31. The method of embodiment 30, wherein the metal hydride is sodium hydride and / or the metal alkoxide is potassium tert-butoxide. E-32. The method of any one of embodiments 1-31, wherein (iv) comprises cooling the mixture at a temperature of about 0°C to room temperature. E-33. The method of any one of embodiments 1 to 32, wherein (v) comprises selectively deprotecting the primary alcohol of Compound (5) to form Compound (6). E-34. The method of any one of embodiments 1 to 33, wherein, in (v), the solution of Compound (5) comprises tetrahydrofuran and a fluoride ion source. E-35. The method of embodiment 34, wherein the fluoride ion source is tetrabutylammonium fluoride. 27Ref: P329WO E-36. The method of any one of embodiments 1 to 35, wherein Compound (6) is selected from , , and . E-37. The method of any one of embodiments 2 and 6 to 36 wherein (vi) comprises reacting Compound (6) with a phosphoryl halide to form Compound (7). E-38. The method of embodiment 37, wherein the phosphoryl halide is phosphoryl trichloride. E-39. The method of any one of embodiments 2 and 6 to 38, wherein (vi) comprises reacting Compound (6) with a phosphoryl halide at a temperature of about 0°C under an inert atmosphere. E-40. The method of any one of embodiments 2 and 6 to 39, wherein (vi) comprises dissolving Compound (6) in a solution comprising triethylamine and diethyl ether. E-41. The method of any one of embodiments 2 and 6 to 40, wherein (vi) comprises adding a solution of Compound (6) to a solution comprising phosphoryl halide dropwise. E-42. The method of any one of embodiments 2 and 6 to 41, wherein (vii) comprises dissolving Compound (7) in a solution comprising diethyl ether and tetrahydrofuran. E-43. The method of embodiment 42, wherein the solution comprises 2- aminoethanol and triethylamine. E-44. The method of any one of embodiments 2 and 6 to 43, wherein (vii) comprises heating the solution at a temperature of about 30°C to 60°C under an inert atmosphere. E-45. The method of embodiment 44, wherein (vii) comprises heating the solution at a temperature of about 50°C with an internal temperature of 40°C. 28Ref: P329WO E-46. The method of any one of embodiments 2 and 6 to 45, wherein the acid in (viii) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate. E-47. The method of any one of embodiments 2 and 6 to 46, wherein, in (viii), the mixture comprises diethyl ether and tetrahydrofuran. E-48. The method of any one of embodiments 2 and 6 to 47, wherein (ix) comprises reacting Compound (9) with at least one catalyst and under an inert atmosphere. E-49. The method of embodiment 48, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. E-50. The method of any one of embodiments 2 and 6 to 49, wherein (ix) comprises reacting Compound (9) in a mixture that is heated at a temperature of about 40°C to 70°C. E-51. The method of embodiment 50, wherein the temperature is about 50°C to 55°C. E-52. The method of embodiment 3, wherein the metal oxide in (vii-a) is silver oxide. E-53. The method of embodiment 3 or embodiment 52, wherein (viii-a) comprises reacting Compound (11) with at least one catalyst and under an inert atmosphere. E-54. The method of embodiment 53, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) 29Ref: P329WO chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. E-55. The method of embodiment 4, wherein (vi-b) comprises reacting Compound (6) with a phosphoryl halide to form Compound (7). E-56. The method of embodiment 55, wherein the phosphoryl halide is phosphoryl trichloride. E-57. The method of any one of embodiment 4, 55, and 56, wherein (vi-b) comprises reacting Compound (6) with a phosphoryl halide at a temperature of about 0°C under an inert atmosphere. E-58. The method of any one of embodiments 4 and 55 to 57, wherein (vi-b) comprises dissolving Compound (6) in a solution comprising triethylamine and diethyl ether. E-59. The method of any one of embodiments 4 and 55 to 58, wherein (vi-b) comprises adding a solution of Compound (6) to a solution comprising a phosphoryl halide dropwise. E-60. The method of any one of embodiments 4 and 55 to 59, wherein (vii-b) comprises dissolving Compound (7) in a solution comprising diethyl ether and tetrahydrofuran. E-61. The method of embodiment 60, wherein the solution comprises ethanolamine and triethylamine. E-62. The method of any one of embodiments 4 and 55 to 61, wherein (vii-b) comprises heating the solution at a temperature of about 30°C to 60°C under an inert atmosphere. E-63. The method of embodiment 62, wherein (vii-b) comprises heating the solution at a temperature of about 50°C with an internal temperature of 40°C. E-64. The method of any one of embodiments 4 and 55 to 63, wherein the acid in (viii-b) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate. E-65. The method of any one of embodiments 4 and 55 to 64, wherein, in (viii- b), the mixture comprises diethyl ether and tetrahydrofuran. 30Ref: P329WO E-66. The method of any one of embodiments 4 and 55 to 65, wherein, in (ix-b), methylating Compound (9) comprises reacting Compound (9) with methyl bromide, methyl iodide, or dimethyl sulfate. E-67. The method of any one of embodiments 4 and 55 to 65, wherein (x-b) comprises reacting Compound (11) with at least one catalyst and under an inert atmosphere. E-68. The method of embodiment 67, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. E-69. The method of embodiment 5, wherein in (vii-c), the oxidising reagent is selected from a peroxy acid or a salt thereof, potassium peroxymonosulfate, and magnesium monoperoxyphthalate. E-70. The method of embodiment 69, wherein the peroxy acid is selected from meta-chloroperoxybenzoic acid and peroxyacetic acid E-71. The method of any one of embodiments 5, 69, or 70, wherein (viii-c) comprises reacting Compound (14) with at least one catalyst and under an inert atmosphere. E-72. The method of embodiment 71, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. E-73. The method of any one of embodiments 5 and 69 to 72, wherein (ix-c) comprises reacting Compound (15) with piperidine to form Compound (10). E-74. A method for synthesizing Compound (12) 31Ref: P329WO , or a salt thereof, the method(1-a) reacting R1-OH with (2R)-(-)-glycidyl tosylate (Compound (18)) to form Compound (19);(19), wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, (1-b) reacting Compound (19) with a benzylating reagent in solution, the benzylating reagent reacting with the secondary alcohol of Compound (19) to form Compound (20); yl,);32Ref: P329WO (1-f) reacting Compound (23) in a solution comprising trimethylamine to form Compound (24); dompound (12) .in the metal acetate is caesium acetate. E-76. A method for synthesizing Compound (10) , or a salt thereof, the method(vi-d) phosphorylating the primary alcohol of Compound (6to form Compound (7);33Ref: P329WO (vii-d) reacting Compound (7) with ethanolamine in solution to form Compound (8); and either[ ] (v -d ) reactng Compound (8) in a mixture comprising acid to form Compound (9)and (ix-d1) deprotecting Compound (9) to form Compound (10); orpound (8a) dixture comprising acid to form Compound (10) ,34Ref: P329WO wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group. E-77. A method for synthesizing Compound (10) , or a salt thereof, the method(vi-e) phosphorylating the primary alcohol of Compound (6to form Compound (7);ethanolamine in solution to form Compound (9); andound (10) ,C30acyl group. 35Ref: P329WO E-78. A method for synthesizing Compound (10) , or a salt thereof, the method(vi-f) phosphorylating the primary alcohol of Compound (6to form Compound (7);(vii-f) reacting Compound (7) with an amino-protected ethanolamine compound in solution to form Compound (9b); , wherein PG is a protecting group,[1] (viii-f1) selectively deprotecting Compound (9b) in a mixture to form Compound (9) d(10); 36Ref: P329WO orn a mixture to form Compound (9c) d(10) ,C30acyl group. E-79. A method for synthesizing Compound (10) , or a salt thereof, the method37Ref: P329WO (vi-f) phosphorylating the primary alcohol of Compound (6to form Compound (7);(vii-f’) reacting a di-protected amino compound HOCH2CH2NPG2 with Compound (7) to yield Compoun , wherein eachPG is independ[1] (viii-f1’) selectively deprotecting Compound (9b’) in a mixture to form Compound (9) d(10); orn a mixture to form Compound (9c’) 38Ref: P329WOand (ix-f2) deprotecting Compound (9c’) to form Compound (10) , wherein R1is a C3-C30alkyl, C3-E-80. A method for synthesizing Compound (10) , or a salt thereof, the method(vi-f) phosphorylating the primary alcohol of Compound (6to form Compound (7);39Ref: P329WO (vii-f’’) reacting a compound HOCH2CH2N+PG3 X- with Compound (7) to yield Compoun , wherein each PG isindepend nion, suc- - - -h as Cl, Br, I, OH, or other suitable counter anion, and either [1] (viii-f1’’) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9) andund (10); orin a mixture to form Compound (9c’’) dorm Compound (10) 40Ref: P329WO, wherein R1is a C3-C30 alkyl, C3- C30 alkenyl, or C3-C30 acyl group. E-81. A method for synthesizing Compound (10) dphosphorodichloridate to form Compound (9a);(vii-g) converting Compound (9a) to Compound (9b); and either) in a mixture to form Compound (9c) 41Ref: P329WO d(10); orin a mixture to form Compound (9) andund (10) ,C30acyl group and PG is a protecting group. E-82. A method for synthesizing Compound (10) 42Ref: P329WO dphosphorodichloridate to form Compound (9a);(vii-g’) converting Compound (9a) to Compound (9b') by reacting a compound HNPG2 with Compound (9a) to yield Compound (9b’), wherein each PG is independently a protecting group; and either [1] (viii-g1) selectively deprotecting Compound (9b’) in a mixture to form Compound (9c’) 43Ref: P329WOand (ix-g1) deprotecting Compound (9c’) to form Compound (10); orb’) in a mixture to form Compound (9) d(10) ,C30acyl group. E-83. A method for synthesizing Compound (10) 44Ref: P329WO dphosphorodichloridate to form Compound (9a);(vii-g’’) converting Compound (9a) to Compound (9b'’) by reacting a compound NPG3 with Compound (9a) to yield Compoun ,wherein each PG is independently a pr[1] (viii-g1) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9c’’) dorm Compound (10); 45Ref: P329WO orin a mixture to form Compound (9) d(10) ,C30acyl group. E-84. A method for synthesizing Compound (10) , or a salt thereof, the method46Ref: P329WO (vi-g) reacting Compound (6)with 2-bromoethyl phosphorodichloridate to form Compound (9a);(vii-g’’’) converting Compound (9a) to Compound (9b'’’) by reacting Compound (9a) with a metal azide to yield Compoun d either[1] (viii-g1) selectively deprotecting Compound (9b’’’) in a mixture to form Compound (9c’’’) andound (10);or 47Ref: P329WO [2] (viii-g2) reducing Compound (9b’’’) to form Compound (9) d(10) , wherein R1is a C3-C30alkyl, C3-E-85. A method for synthesizing Compound (10) dchlorophosphate to form Compound (8b); erRef: P329WO [1] (vii-h1) converting Compound (8b) to Compound (9) dd (10); , ornd (9b),a mixture to form Compound (9c), d(10); 49Ref: P329WOor [3] (vii-h3) converting Compound (8b) to Compound (9b),form Compound (9) dCompound (10) ,C30acyl group and PG is a protecting group. E-86. A method for synthesizing Compound (10) 50Ref: P329WO dchlorophosphate to form Compound (8b); erpound (9) dd (10); , or51Ref: P329WO [2] (vii-h2’) reacting a compound HNPG2 with Compound (8b) to yield Compound (9b’) , wherein each PG is independently a(viii-h2’) selectively deprotecting Compound (9b’) in a mixture to form Compound (9c’), andpound (10); ormpound (8b) to yield Compound (9b’) , wherein each PG is independently a(viii-h3) selectively deprotecting Compound (9b’) to form Compound (9) 52Ref: P329WO andform Compound (10) ,C30acyl group. E-87. A method for synthesizing Compound (10) dchlorophosphate to form Compund (8b); and either8b) to Compound (9) 53Ref: P329WOand (viii-h1) deprotecting Compound (9) to form Compound (10); , orpound (8b) to yield Compound (9b’’), wherein each PG is independently a protecting group; (viii-h2’’) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9c’’), dorm Compound (10);or 54Ref: P329WO [3] (vii-h3’’) reacting a compound NPG3 with Compound (8b) to yield Compound (9b’’) , wherein each PG is independently a protecting(viii-h3) selectively deprotecting Compound (9b’’) to form Compound (9) andform Compound (10) ,C30 acyl group and PG is a protecting group. E-88. A method for synthesizing Compound (10) , or a salt thereof, the method55Ref: P329WO (vi-h) reacting Compound (6)with ethylene glycol chlorophosphate to form Compound (8b); erpound (9) dd (10); , orazide to yield Compound (9b’’’); (viii-h2’’’) selectively deprotecting Compound (9b’’’) in a mixture to form Compound (9c’’’), 56Ref: P329WOand (ix-h2) reducing Compound (9c’’’) to form Compound (10); oride to yield Compound (9b’’’) ;to form Compound (9) dCompound (10) ,57Ref: P329WO wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group. E-89. The method of any one of embodiments 1 to 88, wherein R1is a C16-C18 alkyl, C16-C18 alkenyl, or C16-C18 acyl group. E-90. The method of any one of embodiments 1 to 89, wherein R1is a C16 alkyl group. E-91. The method of any one of embodiments 1 to 89, wherein R1is a C18 alkyl group. E-92. The method of any one of embodiments 1 to 89, wherein R1is a C18 alkenyl group. E-93. The method of any one of embodiments 2, 5-73, and 76-92, wherein Compound (10) is selected from d E-nd 75, wherein Compound (12) is selected from ,58Ref: P329WO d E, or a salt thereof, the method(2-a) providing Compound (26) , wherein each of R1and R3is independently selected from a-C30 alkenyl group, and a C3-C30 acyl group. (2-b) phosphorylating the primary alcohol of Compound (26) to form Compound (27);h ethanolamine in solution to form Compound (28); 59Ref: P329WO dure comprising acid to form Compound (25) .E-96. The method of embodiment 95, wherein R1is selected from a C16-C18 alkyl group, a C16-C18 alkenyl group, and a C16-C18 acyl group. E-97. The method of embodiment 95, wherein R3is an allyl group. E-98. The method of embodiment 97, further comprising removing the allyl group to form Compound (10) , wherein R1is a C3-C30 alkyl, C3-C30E-99. The method of embodiment 95, wherein Compound (25) is further reacted to form Compound(3-a) methylating Compound (25) to form Compound (29); 60Ref: P329WO d12) , wherein R1is a C3-C30 alkyl, C3-C30E-100. A compound selected from Compound (6,Compoun ,Compoun , Compoun ,61Ref: P329WO , , , ,62Ref: P329WO Compound (11, and a salt of any of the foregoing, wherein eaf R1and R3is independently selected from a C16-C18 alkyl group, a C16-C18 alkenyl group, and a C16-C18 acyl group. E-101. The compound of embodiment 100, wherein R1is a C16 alkyl group. E-102. The compound of embodiment 100, wherein R1is a C18 alkyl group. E-103. The compound of embodiment 100, wherein R1is a C18 alkenyl group. E-104. A compound selected from (R)-4-((hexadecyloxy)methyl)-2,2-dimethyl-1,3-dioxolane ;;;;;Ref: P329WO (S,Z)-2-(allyloxy)-3-(octadec-9-en-1-yloxy)propan-1-ol ;;-oxide ;-oxide ;;64Ref: P329WO ( [ ( (; [(2R)-2-allyloxy-3-[(Z)-octadec-9-enoxy]propyl] 2-(trimethylammonio)ethyl phosphate ; and salts thereof. 65Ref: P329WO E-105. The compound or salt thereof of embodiment 104, wherein the compound or salt is not marine-sourced. E-106. A composition comprising one or more compounds or salts thereof of any one of embodiments 100 to 105. E-107. The composition according to embodiment 106, wherein the compound or salt thereof is not marine-sourced. E-108. The compound or salt thereof of any one of embodiments 100 to 105, wherein the compound or salt is produced by the method of any one of embodiments 1 to 99. E-109. A method of using at least one compound selected from Compound (10 ,CompounCompound (9;and / or Compound (11; 66Ref: P329WO wherein the method comprises subjecting Compound (10), Compound (12), Compound (9), and / or Compound (11) to one or more of the following transformations: (A) dehydrogenation of R1, and / or (B) acylation of the secondary alcohol to incorporate an acyl group, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group. E-110. The method of embodiment 109, wherein Compound (10) is subjected to transformation (A) to yield Compound (10A) , wherein R5isup, and a C1-C28 acyl group. E-111. The method of embodiment 109, wherein Compound (10) is subjected to transformation (B) to yield Compound (10B) , wherein R1is selected from aC3-C30 acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group. E-112. The method of embodiment 109, wherein Compound (10) is subjected to transformation (A) and transformation (B) to yield Compound (10AB) 67Ref: P329WO , wherein R5isp, and a C1-C28 acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group. E-113. The method of embodiment 109, wherein Compound (12) is subjected to transformation (A) to yield Compound (12A) , wherein R5isp, and a C1-C28 acyl group. E-114. The method of embodiment 109, wherein Compound (12) is subjected to transformation (B) to yield Compound (12B) , wherein R1is selected froma C3-C30 acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group. E-115. The method of embodiment 109, wherein Compound (12) is subjected to transformation (A) and transformation (B) to yield Compound (12AB) 68Ref: P329WO , wherein R5isp, and a C1-C28 acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group. E-116. The method of embodiment 109, wherein Compound (9) is subjected to transformation (A) to yield Compound (9A) , wherein R5isp, and a C1-C28 acyl group. E-117. The method of embodiment 109, wherein Compound (11) is subjected to transformation (A) to yield Compound (11A), wherein R5is selected from a C1-C28 alkyl group, a C1-C28 alkenyl group, and a C1-C28 acyl group. E-118. The method of any one of embodiments 109-112, wherein compound (10) is formed from the method of any one of embodiments 2, 5-73, and 76-92. E-119. The method of any one of embodiments 109 and 113-115, wherein compound (12) is formed from the method of any one of embodiments 3, 4, 74, and 75. 69Ref: P329WO DETAILED DESCRIPTION

[0060] It should be understood that both the foregoing general description and the following detailed description are exemplary and explanatory but are not restrictive of the invention as claimed. Certain details of one or more embodiments of the invention are set forth in the description below. Those of skill in the art will recognize that there are numerous variations and modifications of the invention that are encompassed by its scope. Other features or advantages of the present disclosure will be apparent from the representative examples that follow, and also from the appended claims. Definitions

[0061] As used herein, nomenclature for compounds including organic compounds, can be given using common names, IUPAC, IUBMB, or CAS recommendations for nomenclature. One of skill in the art can readily ascertain the structure of a compound if given a name, either by systemic reduction of compound structure using naming conventions, or by commercially available software, such as CHEMDRAWTM(Cambridgesoft Corporation, U.S.A.).

[0062] This disclosure includes several numerical values and ranges of numerical values. It is to be understood that each unit between two particular units in a disclosed range are also disclosed. For example, if a range of 10-15 is disclosed, then at least 11, 12, 13, and 14 are also disclosed.

[0063] In the compounds of this disclosure any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as “H” or “hydrogen,” the position is understood to have hydrogen at its natural abundance isotopic composition.

[0064] Aspects of the disclosure also relate to salts of compounds. As used herein, a salt of a compound of this disclosure is formed between an acid and a basic group of the compound, such as an amino functional group, or a base and an acidic group of the compound, such as a carboxyl functional group. In some 70Ref: P329WO embodiments, the compound may be, but is not required to be, a pharmaceutically acceptable salt.

[0065] As used herein, the term “pharmaceutically acceptable” refers to a component that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit / risk ratio. A “pharmaceutically acceptable salt” means any non-toxic salt that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this disclosure.

[0066] Acids commonly employed to form pharmaceutically acceptable salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and phosphoric acid, as well as organic acids such as para-toluenesulfonic acid, salicylic acid, tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, para-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, and acetic acid, as well as related inorganic and organic acids. Such pharmaceutically acceptable salts thus include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hydroxybuterate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1- sulfonate, naphthalene-2-sulfonate, mandelate, and other salts.

[0067] As used herein, the term “alkyl” refers to a saturated straight or branched hydrocarbon, such as a straight or branched group of 1-30, 3-30, 1-18, 14-24, 14-18, 16 or 18 carbon atoms, referred to herein as C1-C30 alkyl, C3-C30 alkyl, C1-C18 71Ref: P329WO alkyl, C14-C24 alkyl, C14-C18 alkyl, C16 alkyl, or C18 alkyl respectively. 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, etc. It will be understood that “alkyl” groups encompass both1H species and deuterated analogues. As used herein, “D” refers to deuterium (2H).

[0068] As used herein, the term “alkenyl” refers to an unsaturated straight or branched hydrocarbon, such as a straight or branched group of 2-30, 3-30, 2-18, 14- 24, 14-18, 16 or 18 carbon atoms, referred to herein as C2-C30 alkenyl, C3-C30 alkenyl, C2-C18 alkenyl, C14-C24 alkenyl, C14-C18 alkenyl, C16 alkenyl, or C18 alkenyl respectively. Multiple double bonds may be present in an alkenyl group. Exemplary alkenyl groups include, but are not limited to, 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, 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, allyl, etc. The term “alkenyl” can additionally be taken to mean an alkene of structure C18:1 wherein the double bond is typically at the n7 or n9 position. The term “alkenyl” can additionally be taken to mean an alkene of structure C18:1 wherein the double bond is between the first and second carbon adjacent to the ether link. The term “alkenyl” can additionally be taken to mean an alkene wherein one double bond is at the n7 or n9 position and a second double bond is between the first and second carbon. It will be understood that “alkenyl” groups encompass both1H species and deuterated analogues.

[0069] As used herein, the term “acyl” refers to a radical of general formula –C(O)R, wherein R is hydrogen or a saturated or unsaturated straight or branched 72Ref: P329WO hydrocarbon. It will be understood that “acyl” groups encompass both1H species and deuterated analogues.

[0070] As used herein, the term “aryl” refers to refers to a functional group or substituent derived from an aromatic ring, such as an aromatic hydrocarbon, such as phenyl. In some embodiments, “aryl” may refer to any substituted or unsubstituted aromatic ring system with 6 to 18 ring carbons. The ring systems may be monocyclic or bicyclic moieties. In some embodiments, “aryl” may refer to any substituted or substituted aromatic ring system with 6 ring carbons. In some embodiments, the substituted ring system with 6 ring carbons is a substituted phenyl group, such as a tolyl group.

[0071] As used herein, the term “mixture” refers to a composition comprising two or more different chemical substances, wherein the individual chemical substances are not chemically bonded. In some embodiments, a mixture may be a solution, a suspension, or a slurry. In some embodiments, a mixture may be homogeneous. In some embodiments, a mixture may be heterogeneous.

[0072] As used herein, the term “solution” refers to a homogeneous mixture. In some embodiments, a solution may be a homogeneous mixture of one or more solutes and one or more solvents.

[0073] As used herein, the term “protecting group reagent” refers to a chemical compound capable of introducing a protecting group onto a nucleophilic functional group, typically by reacting with the nucleophile to form a covalent bond between the protecting group and the nucleophilic atom. The protecting group temporarily masks the reactivity of the nucleophile during subsequent synthetic steps. Non-limiting examples of protecting group reagents for reacting with e.g. alcohols include: tert-butyldimethylsilyl chloride, trimethylsilyl chloride, triethylsilyl chloride, triisopropylsilyl chloride, tert-butyldiphenylsilyl chloride, benzyl halides (e.g., benzyl chloride), and triphenylmethyl halides (e.g., triphenylmethyl chloride).

[0074] As used herein, the term “allylating reagent” refers to a chemical compound capable of introducing an allyl group (–CH₂–CH=CH₂) onto a nucleophilic functional group by forming a covalent bond between the nucleophile and the allyl 73Ref: P329WO group. Non-limiting examples of allylating reagents include allyl halides, such as allyl bromide and allyl chloride.

[0075] As used herein, the term “ammonia surrogate” refers to a chemical compound that can release or transfer an amino group (–NH₂), thereby serving as a functional substitute for ammonia (NH₃) in a chemical transformation, such as a chemical reaction with a substrate of the present disclosure. Ammonia surrogates are typically used to introduce an amino group (–NH₂) into a molecule in place of using free ammonia, which may not be suitable with all reaction conditions. Non-limiting examples of ammonia surrogates of the present disclosure include metal azides (e.g., sodium azide, lithium azide), hydroxylamine, O-benzylhydroxylamine, and ammonium salts such as ammonium chloride, ammonium carbonate, ammonium formate and ammonium acetate. Amines such as hexamethethylene tetramine (urotropine), diallylamine and silylamines may also be condsidered as ammonia surrogates.

[0076] As used herein, the terms “phosphoroyl trichloride”, “phosphoryl trichloride”, “POCl3”, and “phosphoryl trichloride” are used interchangeably herein to refer to phosphoryl trichloride.

[0077] As used herein, the terms “ethylene chlorophosphate”, “2-chloro- 1,3,2-dioxaphospholane 2-oxide”, and “cyclic ethylene phosphate chloride” are used interchangeably herein to refer to 2-chloro-1,3,2-dioxaphospholane 2-oxide.

[0078] As used herein, the term “polar protic solvent” as used herein refers to a solvent (or a mixture of solvents) that is polar, meaning it has a dipole moment, and that contains at least one hydrogen atom bound to a highly electronegative atom such as oxygen or nitrogen (e.g., in –OH or –NH2 groups), allowing the solvent to act as a proton donor in hydrogen bonding. Polar protic solvents are capable of stabilizing charged species via both dipole interactions and hydrogen bonding. Examples of polar protic solvents include, but are not limited to, water, methanol, ethanol, propan-1-ol, propan-2-ol, and higher alcohols.

[0079] As used herein, the term “polar aprotic solvent” as used herein refers to a solvent (or a mixture of solvents) that is polar, meaning it has a dipole moment, but lacks acidic protons (e.g., in –OH or –NH2 groups) and therefore cannot act as a proton donor in hydrogen bonding. Examples of polar aprotic solvents include, 74Ref: P329WO but are not limited to, acetone, methyl ethyl ketone (MEK), methyl isobutyl ketone (MIBK) dimethyl formamide, dimethyl acetamide, N-methyl pyrrolidone, dimethyl sulfoxide, sulfolane, acetonitrile and dichloromethane.

[0080] As used herein the term “aprotic ethereal solvent” refers to a solvent (or a mixture of solvents) that is aprotic, meaning it lacks hydrogen atoms bound to highly electronegative atoms (e.g., no –OH or –NH₂ groups), and that contains one or more ether functionalities (–C–O–C–) as part of its structure. Examples of aprotic ethereal solvents include, but are not limited to, diethyl ether, methyl tert-butyl ether (MTBE), tetrahydrofuran (THF), 1,4-dioxane, cyclopentyl methyl ether (CPME), dimethyl ether and diglyme, for example diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, 2-methyl tetrahydrofuran, and cyclopentyl methyl ether.

[0081] As used herein, the terms “C16.0 analogue”, “C18.0 analogue” and “C18.1 analogue” refer to compounds of the present disclosure, possibly including reference to a compound number, such as “C18.0 analogue of Compound (7)”, wherein the R group or R1 group is a straight alkyl chain which for C16.0 analogue is C16H33 (hexadecanyl); for C18.0 analogue is C18H37 (octadecanyl), and for C18.0 analogue is C18H35 ((Z)-octadec-9-en-1-yl).

[0082] As used herein, the terms “BOC”, “BoC”, and “Boc” are used interchangeably referring to the tert-butoxycarbonyl group. Synthesis of Compound (6)

[0083] In some embodiments, the present disclosure concerns the synthesis of Compound (6), an important intermediate in the synthesis of e.g. plasmalogen precursors, and derivatives thereof.

[0084] In some embodiments, the disclosure provides methods for synthesizing chemical precursors to bioactive compounds. In some embodiments, the disclosure provides a method for synthesizing Compound (6) or a salt thereof .75Ref: P329WO

[0085] In some embodiments, the method comprises (i) reacting (R)-(2,2- dimethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1-OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2) .

[0086] In someembodiments, the method comprises (i) reacting Compound (1) and the R1-OSO2R2or the R1-X in the presence of a base. In some embodiments, the base may be an organic base. In some embodiments, the base may be an inorganic base. In some embodiments, the inorganic base may be an alkali metal base. In some embodiments, the alkali metal base may be an alkali metal hydroxide, an alkali metal alkoxide, or an alkali metal carbonate. In some embodiments, the alkali metal hydroxide, alkali metal alkoxide, or alkali metal hydride may be, but is not limited to, selected from sodium hydroxide, potassium hydroxide, potassium butoxide, sodium tert-butoxide, potassium tert-butoxide, sodium hydride, and potassium hydride. In some embodiments, the base is potassium hydroxide. In some embodiments, the base may be a non-nucleophilic alkoxide base.

[0087] In some embodiments, the base is potassium tert-butoxide.

[0088] In some embodiments, from 1 to 4 equivalents of the base is used, such as from 1 to 3, such as about 2 equivalents.

[0089] In some embodiments, (i) is conducted in an apolar solvent or a polar solvent, for example wherein (i) is conducted in toluene.

[0090] In some embodiments, (i) comprises reacting Compound (1) for at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, such as at least 2 hours, for example from 15 minutes to 48 hours.

[0091] In some embodiments, (i) further comprises adding a suitable reagent to the solution. In some embodiments, the suitable reagent may be, but is not 76Ref: P329WO limited to, a quaternary ammonium salt. In some embodiments, the suitable reagent is tetrabutylammonium bromide. In some embodiments, the addition of tetrabutylammonium bromide occurs before, during, or after the addition of the R1- OSO2R2or the R1-X. In some embodiments, the addition of tetrabutylammonium bromide occurs after the addition of the R1-OSO2R2or the R1-X.

[0092] In some embodiments, (i) comprises reacting Compound (1) with R1- X, for example wherein X is bromide, and optionally wherein R1is a C10-C24 alkyl or C10-C24 alkenyl.

[0093] In some embodiments, (i) comprises reacting Compound (1) with the R1-OSO2R2or R1-X at room temperature. In some embodiments, (i) further comprises heating the solution. In some embodiments, (i) comprises heating the solution at a temperature range of about 60°C to 100°C. In some embodiments, (i) comprises heating the solution at a temperature range of about 40°C to 120°C, about 50°C to 110°C, about 60°C to 100°C, about 70°C to 90°C, or about 75°C to 85°C. In some embodiments, (i) comprises heating the solution at a temperature of about 80°C.

[0094] In some embodiments, (i) further comprises purifying Compound (2) before hydrolyzing Compound (2). In some embodiments, in (i), Compound (2) is not purified before hydrolyzing Compound (2).

[0095] In some embodiments, the method further comprises (ii) hydrolyzing Compound (2) in a mixture comprising acid to form Compound (3) .

[0096] In some e(ii) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate.

[0097] In some embodiments, (ii) further comprises dissolving Compound (2) in a solvent before hydrolyzing Compound (2).

[0098] In some embodiments, the solvent in (ii) is a polar aprotic solvent, a polar protic solvent, or a mixture thereof. 77Ref: P329WO

[0099] In some embodiments, the solvent is selected from the group consisting of: tetrahydrofuran, diethyl ether, and 2-methyl tetrahydrofuran, for example tetrahydrofuran.

[0100] In some embodiments, the solvent in (ii) is an aprotic ethereal solvent, such as an aprotic ethereal solvent selected from the group consisting of: diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, 2-methyl tetrahydrofuran, cyclopentyl methyl ether.

[0101] In some embodiments, the solvent is a polar protic solvent, for example methanol.

[0102] In some embodiments, the solution is heated at from 50 °C to 70 °C, such as about 60 °C.

[0103] In some embodiments, (ii) comprises reacting Compound (2) for at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, such as at least 2 hours, for example from 15 minutes to 48 hours.

[0104] In some embodiments, the solvent may be, but is not limited to, an organic solvent. In some embodiments, the organic solvent may be, but is not limited to, toluene, hexane, chloroform, ethyl acetate, tetrahydrofuran, methylene chloride, diethyl ether, or another suitable solvent. In some embodiments, the organic solvent is tetrahydrofuran.

[0105] In some embodiments, (ii) further comprises heating the solution. In some embodiments, (ii) comprises heating the mixture at a temperature range of about 50°C to 80°C. In some embodiments, (i) comprises heating the solution at a temperature range of about 30°C to 100°C, about 40°C to 90°C, about 50°C to 80°C, or about 60°C to 70°C. In some embodiments, (i) comprises heating the solution at a temperature of about 65°C.

[0106] In some embodiments, (ii) further comprises extracting Compound (3) from the mixture. In some embodiments, the extraction is performed with 2- methyltetrahydrofuran, tetrahydrofuran, dichloromethane, or another suitable extraction solvent. In some embodiments, the extraction is performed with 2- methyltetrahydrofuran. 78Ref: P329WO

[0107] In some embodiments, (ii) comprises extracting Compound (3) from the mixture with an aprotic solvent, such as an ethereal aprotic solvent, for example as disclosed herein, such as 2-methyltetrahydrofuran.

[0108] In some embodiments, (ii) comprises reacting with HCl in methanol at for example from 50 °C to 70 °C, such as 60 °C over the course of for example from 4 hours to 18 hours, such as 6 hours.

[0109] In some embodiments, (ii) comprises reacting with from 0.05M to 2.00 M HCl, such as from 0.1M to 1.50M, for example from 0.1M to 0.5M HCl, optionally using from 1 to 10 Volumes, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 Volumes of HCl.

[0110] In some embodiments, the method further comprises (iii) reacting Compound (3) with a protecting group reagent in solution, the protecting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4) , wherein PG is a protecting g

[0111] Protecting groups suitable for use according to the disclosure, and their introduction and removal, are familiar to persons having ordinary skill in the art and include, for example, silicon- and carbon-based functional groups. Non-limiting examples of protecting groups include silicon-containing groups, such as tert- butyldimethylsilyl, trimethylsilyl, triethylsilyl, and triisopropylsilyl groups, and carbon- containing groups, such as benzyl, p-methoxybenzyl, and triphenylmethyl (trityl) groups. As used herein, a protecting group reagent is a chemical reagent suitable for incorporating a suitable protecting group into a chemical structure. Protecting group reagents suitable for use according to the disclosure are also familiar to persons having ordinary skill in the art and include, for example, chemical reagents suitable for incorporating silicon- or carbon-based functional groups into a chemical structure. Non- limiting examples of protecting group reagents include tert-butyldimethylsilyl chloride, trimethylsilyl chloride, triethylsilyl chloride, triisopropylsilyl chloride, a benzyl halide, such as benzyl bromide, benzyl chloride, p-methoxybenzyl bromide, or p-methoxybenzyl chloride, and a trityl halide, such as trityl chloride. 79Ref: P329WO

[0112] In some embodiments, the protecting group reagent in (iii) is selected from tert-butyldimethylsilyl chloride, trimethylsilyl chloride, triethylsilyl chloride, triisopropylsilyl chloride,tert-butyl diphenyl silyl chloride, a benzyl halide, and a triphenylmethyl halide. Other suitable protecting groups and protecting group reagents are known to persons having ordinary skill in the art. See, e.g., Peter G.M. Wuts, Greene’s Protective Groups in Organic Synthesis, 5th Edition, 2014, the contents of which is incorporated by reference herein. In some embodiments, in (iii) the solution comprises one or more of pyridine, dichloromethane, imidazole, and dimethylformamide. In some embodiments, in (iii) the solution comprises two or more of or three or more of pyridine, dichloromethane, imidazole, and dimethylformamide. In some embodiments, in (iii) the solution comprises pyridine, dichloromethane, and imidazole. In some embodiments, in (iii) the solution comprises pyridine, dichloromethane, imidazole, and dimethylformamide.

[0113] In some embodiments, in (iii), the temperature of the solution is maintained at from about 0°C to room temperature.

[0114] In some embodiments, in (iii), the temperature of the solution is maintained at from -10 °C to 40 °C, such as from about 0°C to room temperature, for example between about 25 and 30 °C.

[0115] In some embodiments, in (iii), the solution comprises a polar solvent, such as a polar aprotic or polar protic solvent.

[0116] In some embodiments, the polar aprotic solvent in (iii) is selected from the group consisting of: dimethyl formamide, dimethyl acetamide, N-methyl pyrrolidone, dimethyl sulfoxide, sulfoilane, acetonitrile and dichloromethane.

[0117] In some embodiments, the solution in (iii) comprises an ethereal solvent, for examble an ethereal aprotic solvent as disclosed herein, such as THF, 2- MeTHF, or diethyl ether, for example THF.

[0118] In some embodiments, the protecting group reagent in (iii) is added in a stoichiometry of from 0.9 to 3.0 equivalents, such as from 1.0 to 2.5 equivalents, for example from 1.0 to 2.0 equivalents, relative to the Compound (2). 80Ref: P329WO

[0119] In some embodiments, in (iii), a base, such as a nucleophilic base is added to the solution, for example a base capable of electrophilic activation of the protecting group reagent.

[0120] In some embodiments, the base is selected from the group consisting of: imidazole, pyridine, 4-DMAP, triethylamine, N-methylimidazone, and 2,6- lutidine.

[0121] In some embodiments, from 2 to 8 equivalents of the base is added to the solution relative to Compound (2), such as from 2 to 7 equivalents, such as from 3 to 6 equivalents, for example from 4 to 5 equivalents.

[0122] In some embodiments, the method further comprises (iv) reacting Compound (4) in a mixture comprising an allylating reagent to form Compound (5).

[0123] As used herein, an allylating reagent is a chemical reagent suitable for incorporating an allyl group (−CH2−HC=CH2) into a chemical structure. Allylating reagents suitable for use according to the disclosure are familiar to persons having ordinary skill in the art and include, for example, allyl halides.

[0124] In some embodiments, the allylating reagent in (iv) is an allyl halide, such as allyl bromide or allyl chloride. In some embodiments, the allyl halide may be, but is not limited to, allyl bromide.

[0125] In some embodiments, in (iv), the mixture comprises dry tetrahydrofuran.

[0126] In some embodiments, in (iv), the mixture comprises essentially dry tetrahydrofuran, for example having a water content determined by Karl Fisher of from 0.0 ppm to 80 ppm, such as from 0.0 ppm to 70 ppm, such as from 0.0 ppm to 60 ppm, such as from 0.9 ppm to 50 ppm.

[0127] In some embodiments, in (iv), the mixture comprises an apolar solvent, for example a water immiscible solvent, such as toluene.

[0128] In some embodiments, in (iv), the mixture comprises toluene, THF, MTBE, DMAc, n-heptane, and / or acetonitrile. 81Ref: P329WO

[0129] In some embodiments, in (iv), the mixture comprises a non- nucleophilic base, for example a tert-butoxide.

[0130] In some embodiments, in (iv), the mixture comprises a base which is a metal hydride and / or a metal alkoxide.

[0131] In some embodiments, in (iv), the mixture comprises a metal hydride and / or a metal alkoxide. In some embodiments, the metal hydride may be, but is not limited to, an alkali metal hydride. In some embodiments, the metal hydride is sodium hydride. In some embodiments, the metal hydride is potassium hydride. In some embodiments, the metal alkoxide may be, but is not limited to, an alkali metal alkoxide. In some embodiments, the metal alkoxide is sodium butoxide or sodium tert- butoxide. In some embodiments, the metal alkoxide is potassium butoxide or potassium tert-butoxide.

[0132] In some embodiments, (iv) further comprises cooling the mixture at a temperature of about 0°C to room temperature.

[0133] In some embodiments, (iv) comprises cooling the mixture at a temperature of from -10 °C to 50 °C, such as from about 0°C to room temperature, for example between about 25 °C to about 45 °C.

[0134] In some embodiments, in (iv), from 1.0 to 2.5 equivalents of the base is used, such as from 1.1 to 2.0 equivalents, for example from 1.2 to 1.8 equivalents, for example about 1.5 equivalents.

[0135] In some embodiments, in (iv), from 1.0 to 2.5 equivalents of the allylating reagent is used, such as from 1.1 to 2.0 equivalents, for example from 1.2 to 1.8 equivalents, for example about 1.5 equivalents.

[0136] In some embodiments, the method further comprises (v) deprotecting Compound (5) in solution to form Compound (6) .

[0137] For examts, (v) comprises selectively deprotecting the primary alcohol of Compound (5) to form Compound (6). 82Ref: P329WO

[0138] In some embodiments, in (v), the solution of Compound (5) comprises tetrahydrofuran and a fluoride anion source.

[0139] As used herein a fluoride ion source is a chemical reagent that provides a source of fluoride ions in a chemical reaction. The fluoride ion source may be suitable for deprotection of a silicon-containing protecting group described elsewhere herein. The fluoride ion source may be a chemical reagent that provides a source of fluoride anions in a chemical reaction. Non-limiting examples of fluoride ion sources include tetrabutylammonium fluoride, pyridine-(HF)x, triethylamine trihydrofluoride, hydrofluoric acid (HF), tris(dimethylamino)sulfonium difluorotrimethylsilicate, and ammonium fluoride. In some embodiments, the fluoride anion source is tetrabutylammonium fluoride.

[0140] In some embodiments, in (v), a Lewis acid is added to the mixture, for example a Lewis acid selected from the group consisting of: FeCl3, TiCl4, BCl3, ZnBr₂, ZnCl₂, AlCl₃, BF₃·Et₂O, Sc(OTf)₃, InCl₃, and Cu(OTf)₂. The Lewis acids of the present disclosure may be complexed with one or more ligands. In some emodiments, boron halide reagents of the present disclosure are complexed with for example Et2O, Me2S or other suitable ligands.

[0141] In some embodiments, Compound (6) is selected from ,, and .

[0142] In some embodiments, R1 is selected from the group consisting of: hexadecanyl; octadecanyl, and (Z)-octadec-9-en-1-yl. Unless specified otherwise, the R1 group is unbranched and attached to O by position 1. 83Ref: P329WO Synthesis of Compound (10)

[0143] In some embodiments, the disclosure provides methods for synthesizing bioactive in vivo precursors to plasmalogen compounds. In some embodiments, the disclosure provides methods for synthesizing LPE(O)s. In some embodiments, the disclosure provides a method for synthesizing Compound (10) (depicted below in a non-ionic form) or a salt thereof .

[0144] (i) reacting (R)-(2,2- dimethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1-OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2) .

[0145] In some embodiments, the method further comprises (ii) hydrolyzing Compound (2) in a mixture comprising acid to form Compound (3) .

[0146] In some ed further comprises (iii) reacting Compound (3) with a protecting group reagent in solution, the protecting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4) 84Ref: P329WO .

[0147] In somefurther comprises (iv) reacting Compound (4) in a mixture comprising an allylating reagent to form Compound (5).

[0148] In some embodiments, the method further comprises (v) deprotecting Compound (5) in solution to form Compound (6).

[0149] One of ordinary skill in the art will appreciate that the reaction conditions suitable for (i) to (v) are as described above with reference to the synthesis of Compound (6).

[0150] In some embodiments, the method further comprises (vi) phosphorylating the primary alcohol of Compound (6) to form Compound (7).

[0151] In some embodiments, (vi) comprises reacting Compound (6) with a phosphoryl halide to form Compound (7). In some embodiments, (vi) comprises reacting Compound (6) with a phosphoryl halide at a temperature of about 0°C under an inert atmosphere. In some embodiments, the phosphoryl halide is phosphoryl trichloride. 85Ref: P329WO

[0152] In some embodiments, (vi) comprises reacting Compound (6) with a phosphoryl halide at a temperature of from -20 °C to 30 °C, such as from -10 °C to 10 °C for example about 0°C under an inert atmosphere.

[0153] As used herein, an inert atmosphere is a gaseous atmosphere under which a chemical reaction is performed that prevents the ingress of atmospheric oxygen and moisture and does not impede the chemical reaction being performed. Non- limiting examples of gases used to create an inert atmosphere include nitrogen and argon. In the present context, an inert atmosphere is understood to be established by applying positive pressure of an inert gas, such as nitrogen or argon, to the reaction vessel, optionally after one or more evacuation cycles alternated with reapplication of the inert gas.

[0154] In some embodiments, in (vi), from 1.0 to 3.0 equivalents of the phosphoryl halide is used, such as from 1.2 equivalents to 2.8 equivalents, such as from 1.4 equivalents to 2.6 equivalents, for example from 1.6 equivalents to 2.4 equivalents, for example about 2.0 equivalents.

[0155] In some embodiments, (vi) comprises reacting Compound (6) in a apolar solvent, such as toluene, optionally using from 0.2 volumes to 5 volumes of toluene relative to Compound (6).

[0156] In some embodiments, (vi) comprises reacting Compound (6) over the course of from 4 hours to 48 hours, such as from 6 hours to 42 hours, such as from 8 hours to 36 hours, such as from 10 hours to 30 hours, for example from 12 hours to 22 hours, for example about 16 hours.

[0157] In some embodiments, (vi) comprises adding a solution of Compound (6) to a solution comprising phosphoryl halide dropwise.

[0158] In some embodiments, (vi) further comprises dissolving Compound (6) in a solution comprising triethylamine and diethyl ether.

[0159] In some embodiments, (vi) comprises adding a solution of Compound (6) to a solution comprising phosphoryl halide dropwise.

[0160] In some embodiments, the method further comprises (vii) reacting Compound (7) with ethanolamine in solution to form Compound (8) 86Ref: P329WO .

[0161] In soided for production of Compound (10) or Compound (12) disclosed herein, or derivatives thereof disclosed in “Further Transformations” herein, said method comprising reacting Compound (8) in a mixture comprising acid to form Compound (9), and further reaction Compound (9) with a method disclosed herein to provide Compound (10) or Compound (12).

[0162] In some embodiments, the method further comprises reacting Compound (7) with ethanolamine in solution to form Compound (9). In some embodiments, the method comprises a further step of hydrolysis subsequent to addition of ethanolamine to Compound (7).

[0163] In some embodiments, (vii) further comprises dissolving Compound (7) in a solution comprising diethyl ether and tetrahydrofuran.

[0164] In some embodiments, (vii) comprises dissolving Compound (7) in a solution comprising a polar aprotic solvent, for example diethyl ether, tetrahydrofuran, 2-MeTHF, or a mixture thereof.

[0165] In some embodiments, the solution further comprises 2- aminoethanol and triethylamine.

[0166] In some embodiments, from 1.0 to 3.0 equivalents of 2- aminoethanol is used, such as from 1.2 equivalents to 2.8 equivalents, such as from 1.4 equivalents to 2.6 equivalents, for example about 1.5 equivalents.

[0167] In some embodiments, from 1.5 to 4.0 equivalents of the amine base is used, such as from 1.8 equivalents to 3.7 equivalents, such as from 2.0 equivalents to 3.5 equivalents, for example about 3.0 equivalents, optionally wherein the amine base is triethylamine.

[0168] In some embodiments, in (vii), the temperature is from -20 °C to 30 °C, such as from -15 °C to 25 °C, for example from -10 °C to 20 °C, optionally from -10 °C to 10 °C, for example about 0 °C. 87Ref: P329WO

[0169] In some embodiments, in (vii), Compound (7) is reacted for from 5 minutes to 24 hours, such as from 10 minutes to 14 hours, such as from 15 minutes to 8 hours, such as from 30 minutes to 6 hours, such as from 45 minutes to 4 hours, for example about 1 hour.

[0170] In some embodiments, (vii) further comprises heating the solution. In some embodiments, (vii) comprises heating the solution under an inert atmosphere. In some embodiments, (vii) comprises heating the solution at a temperature range of about 30°C to 60°C under an inert atmosphere. In some embodiments, (vii) comprises heating the solution at a temperature range of about 25°C to 70°C, about 30°C to 60°C, or about 40°C to 50°C. In some embodiments, (vii) comprises heating the solution at a temperature of about 50°C with an internal temperature of 40°C under an inert atmosphere.

[0171] In some embodiments, the method further comprises (viii) reacting Compound (8) in a mixture comprising acid to form Compound (9).

[0172] In some embodiments, the acid in (viii) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, a carboxylic acid, a sulfonic acid, and an alkali metal dihydrogen phosphate. In some embodiments, the acid in (viii) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate. In some embodiments, the acid in (viii) is hydrochloric acid.

[0173] In some embodiments, in (viii), the mixture comprises diethyl ether and tetrahydrofuran.

[0174] In some embodiments, (viii) comprises adding an acid, such as an aqueous acid, for example aqueous HCl to the mixture. 88Ref: P329WO

[0175] In some embodiments, the solution and any precipitate therein in (vii) subsequent to mixing of 2-aminoethanol and Compound (7) is subjected to filtration to obtain a filtrate used directly in step (viii).

[0176] In some embodiments, step (viii) further comprises a purification step subsequent to the addition of the acid, wherein the purification step comprises: a. subjecting a composition of Compound (9) to a solvent system, b. heating the solvent system to a predefined temperature for a predefined period of time, c. allowing the solvent system to cool to a temperature lower than the predefined temperature thereby obtaining a composition of Compound (9) having increased purity relative to the composition of Compound (9) in step a.

[0177] In some embodiments, the predefined temperature is from 30 °C to 100 °C, such as from 35 °C to 90 °C, such as from 40 °C to 80 °C, such as from 45 °C to 70 °C, for example from 50 °C to 65 °C, for example about 60 °C.

[0178] In some embodiments, the predefined period of time is at least 15 minutes, such as at least 30 minutes, for example from 15 minutes to 48 hours, for example about 1 hour.

[0179] In some embodiments, the solvent system comprises a polar aprotic solvent or a mixture of different polar aprotic solvents.

[0180] In some embodiments, the solvent system comprises a solvent selected from the group consisting of: MeCN, MTBE, 2-MeTHF, acetone, IPAc, and a mixture thereof.

[0181] In some embodiments, the solvent system comprises 2-MeTHF and acetone, for example in a mixture of 2-MeTHF and acetone with from 10% to 50% 2- MeTHF, the balance being acetone, for example about 1:2 v / v 2-MeTHF:acetone.

[0182] In some embodiments, the crude composition of Compound (9) is added to a first solvent, such as an ethereal solvent, for example 2-MeTHF, after which a second solvent is added to the first solvent, for example acetone.

[0183] In some embodiments, the method further comprises (ix) deprotecting Compound (9) to form Compound (10) 89Ref: P329WO .

[0184] r producing Compound (10), said method comprising deprotecting Compound (9) to form Compound (10), optionally using a method as defined herein, such as reaction step (ix).

[0185] In some embodiments, (ix) comprises reacting Compound (9) with at least one catalyst. In some embodiments, (ix) comprises reacting Compound (9) with at least one catalyst and under an inert atmosphere.

[0186] Catalysts suitable for deprotecting compounds of the disclosure are familiar to persons having ordinary skill in the art. Non-limiting examples of suitable catalysts for deprotecting compounds of the disclosure include palladium, rhodium, and nickel catalysts, including palladium(0) catalysts, palladium (II) catalysts, and rhodium(I) catalysts. In some embodiments, a suitable catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. In some embodiments, a catalyst derived from any of the foregoing or generated in situ from any of the foregoing by combining any of the foregoing with a ligand, such as a phosphine ligand, such as triphenylphosphine, tri-tert-butylphosphine, or 1,1′- ferrocenediyl-bis(diphenylphosphine). In some embodiments, the catalyst is tetrakis(triphenylphosphine)palladium(0). However, one of ordinary skill in the art would also understand that alternate deprotection catalysts may be suitable for the removal of an allyl protecting group. See, e.g., Peter G.M. Wuts, Greene’s Protective Groups in Organic Synthesis, 5th Edition, 2014, the contents of which is incorporated herein by reference.

[0187] In some embodiments, (ix) further comprises reacting Compound (9) in a mixture. In some embodiments, (ix) comprises reacting Compound (9) in a 90Ref: P329WO mixture that is heated at a temperature range of about 40°C to 70°C. In some embodiments, (ix) comprises reacting Compound (9) in a mixture that is heated at a temperature range of about 25°C to 90°C, about 30°C to 80°C, about 40°C to 70°C, about 50°C to 60°C, or about 50°C to 55°C. In some embodiments, (ix) comprises reacting Compound (9) in a mixture that is heated to a temperature range of about 50°C to 55°C.

[0188] In some embodiments, (ix) comprises reacting Compound (9) with a Lewis acid. In some embodiments, the Lewis acid used in step (ix) is as defined elsewhere herein.

[0189] In some embodiments, the Lewis acid comprises boron, such as a boron halide.

[0190] In some embodiments, the Lewis acid is selected from the group consisting of: BF3, BCl3, and BBr3.

[0191] In some embodiments, the Lewis acid is selected from the group consisting of: BF3·Et2O. and BCl3·DCM, for example wherein the Lewis acid is BCl3·DCM.

[0192] In some embodiments, (ix) comprises adding a ligand to the Lewis acid, for example wherein the ligand is a sulfide-based ligand, such as dimethylsulfide, thioanisole, and thiosalicylic acid, for example wherein the ligand is dimethylsulfide.

[0193] In some embodiments, from 1.5 to 6.0 equivalents of the Lewis acid is used in (ix), such as from 2.0 equivalents to 4.5 equivalents, such as from 2.5 equivalents to 4.0 equivalents, for example about 3.5 equivalents.

[0194] In some embodiments, from 1.5 to 6.0 equivalents of the ligand is used in (ix), such as from 2.0 equivalents to 4.5 equivalents, such as from 2.5 equivalents to 4.0 equivalents, for example about 3.5 equivalents or about 4.0 equivalents.

[0195] In some embodiments, from 2.0 equivalents to 4.5 equivalents of the Lewis acid and ligand is used, for example about 3.5 equivalents for the Lewis acid and the ligand or about 4.0 equivalents.

[0196] In some embodiments, (ix) is conducted in a polar aprotic solvent, such as a chlorinated solvent, for example dichloromethane. 91Ref: P329WO

[0197] In some embodiments, in (ix), the reaction is conducted for at least 1 hour, such as from 1 hour to 1 week.

[0198] In some embodiments, (ix) further comprises a silane, such as a silane selected from the group consisting of: triethylsilane, phenylsilane, diphenylsilane, polymethylhydrosiloxane (PMHS), triisopropylsilane, trimethoxysilane, dimethoxymethylsilane, and tetramethyldisiloxane (TMDS).

[0199] In some embodiments, from 2.0 to 8.0 equivalents of the silane is used, for example from 3.0 to 7.0, such as about 6.0 equivalents.

[0200] In some embodiments, the temperature in (ix) is from 0 °C to 80 °C, such as from 10 °C to 70 °C, such as from 15 °C to 60 °C, for example from about 20 °C to about 50 °C, for example about 20 °C to 40 °C.

[0201] In some embodiments, the method comprises reacting bis(2- cyanoethyl)-N,N-diisopropylphosphoramidite with Fmoc-protected aminoethanol to give the corresponding diisopropylphosphoramidite Compound (16) 6) and reacting Compound (1ompound (13).

[0202] In some embodiments, the method further comprises (vii-c) reacting Compound (13) with an oxidizing reagent to form Compound (14) 92Ref: P329WO.

[0203] Oxidizing reagents suitable for use in methods of the disclosure are familiar to persons having ordinary skill in the art. Non-limiting examples of suitable oxidizing reagents include a peroxy acid or a salt thereof, potassium peroxymonosulfate, and magnesium monoperoxyphthalate. In some embodiments, the oxidizing reagent in (vii-c) is selected from a peroxy acid or a salt thereof, potassium peroxymonosulfate, and magnesium monoperoxyphthalate. In some embodiments, the peroxy acid is selected from meta-chloroperoxybenzoic acid and peroxyacetic acid.

[0204] In some embodiments, the method further comprises (viii-c) selectively deprotecting Compound (14) at the secondary alcohol to form Compound (15) .

[0205] g Compound (14) with at least one catalyst. In some embodiments, (viii-c) comprises reacting Compound (14) with at least one catalyst and under an inert atmosphere.

[0206] In some embodiments, in (viii-c), the catalyst may be a palladium(0) catalyst, a palladium(II) catalyst, or a rhodium (I) catalyst. In some embodiments, the catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. In some embodiments, a 93Ref: P329WO catalyst derived from any of the foregoing or generated in situ from any of the foregoing by combining any of the foregoing with a ligand, such as a phosphine ligand, such as triphenylphosphine, tri-tert-butylphosphine, or 1,1′-ferrocenediyl-bis(diphenylphosphine). However, one of ordinary skill in the art would also understand that alternate deprotection catalysts may be suitable for the removal of an allyl protecting group as described elsewhere herein.

[0207] In some embodiments, the method further comprises (ix-c) deprotecting Compound (15) to form Compound (10) .

[0208] g Compound (15) with piperidine to form Compound (10).

[0209] The disclosure provides alternate methods for synthesizing Compound (10). In some embodiments, the method comprises (vi-d) phosphorylating the primary alcohol of Compoun to form Compound (7).

[0210] In some embodiments, the method further comprises (vii-d) reacting Compound (7) with ethanolamine in solution to form Compound (8) 94Ref: P329WO .

[0211] In soer comprises either [1] (viii- d1) reacting Compound (8) in a mixture comprising acid to form Compound (9)and (ix-d1) deprotecting Compound (9) to form Compound (10) ; or [2] (viii-d2) deand (ix-d2) reacting Comacid, such as a weak acid, to form Compound (10) 95Ref: P329WO .

[0212] n one ano er em o men, e me o comprises (vi-e) phosphorylating the primary alcohol of Compound (6to form Compound (7).

[0213] In some embodiments, the method further comprises (vii-e) reacting Compound (7) with ethanolamine in solution to form Compound (9).

[0214] In some embodiments, the method further comprises (viii-e) deprotecting Compound (9) to form Compound (10) .96Ref: P329WO

[0215] In some embodiments, the method comprises (vi-f) phosphorylating the primary alcohol of Compoun to form Compound (7).

[0216] In some embodiments, the method further comprises (vii-f) reacting Compound (7) with an amino-protected ethanolamine (e.g., HOCH2CH2NHPG) compound in solution to form Compound (9b) , wherein PG is a protecting group.), the amino-protected ethanolamine compound is selected from the group consisting of: a. 2-(N-tert-butoxycarbonylamino)ethanol, to provide Compound (9b), wherein PG is Boc; b. benzyl ethanolamine, to provide Compound (9b), wherein PG is benzyl; c. 2-[(4-methoxybenzyl)amino]ethanol, to provide Compound (9b), wherein PG is 4-methoxybenzyl; d. 2-(allylamino)ethanol, to provide Compound (9b), wherein PG is allyl; and e. N-tritylethanolamine, to provide Compound (9b), wherein PG is trityl.

[0218] In some embodiments, in (vii-f), a base is added to the solution, such as an amine base or a metal carbonate.

[0219] In some embodiments, the amine base is triethylamine.

[0220] In some embodiments, in (vii-f) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (7), such as from 1.5 97Ref: P329WO equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

[0221] In some embodiments, in (vii-f) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

[0222] In some embodiments, in (vii-f) the temperature is from -10 °C to 80 °C, such as from 0 °C to 70 °C, for example from 10 °C to 60 °C.

[0223] In some embodiments, in (vii-f) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

[0224] In some embodiments, the method further comprises either [1] (viii- f1) selectively deprotecting Compound (9b) in a mixture to form Compound (9)and (ix-f1) deprotecting Compound (9) to form Compound (10) or [2] (viii-f2) seleform Compound (9c) 98Ref: P329WO , wherein PG is a protecting group,mpound (10) .

[0225] (vi-g) reacting Compound (6)with 2-bromoethyl phosphorodichloridate to form Compounda).

[0226] In some embodiments, the method further comprises (vii-g) converting Compound (9a) to Compound (9b), wherein PG is a protecting group. 99Ref: P329WO

[0227] In some embodiments, (vii-g) comprises reacting Compound (9a) with a protected amine or an ammonia derivative having a protecting group such as a benzyl or allyl protecting group, such as benzylamine, 4-methoxybenzylamine, or allylamine, followed by removal of the protecting group.

[0228] In some embodiments, in (vii-g), Compound 9a is reacted with a protected amine (PGNH2) selected from the group consisting of: a. benzylamine to provide Compound (9b), wherein PG is benzyl; b. 4-methoxybenzyl to provide Compound (9b), wherein PG is 4- methoxybenzyl; and c. Allylamine, for example allylamine·HCl, to provide Compound (9b), wherein PG is allyl.

[0229] In some embodiments, in (vii-g) from 1.0 to 5.0 equivalents of the protected amine is used, such as from 1.2 equivalents to 5.0 equivalents, for example from 1.5 equivalents to 4.0 equivalents, such as about 3.0 equivalents or such as about 4.0 equivalents.

[0230] In some embodiments, in (vii-g) a base is used, such as an amine base. In some embodiments, the amine base is triethylamine.

[0231] In some embodiments, in (vii-g) Compound (9a) is reacted with the protected amine in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

[0232] In some embodiments, in (vii-g) the temperature is from 10 °C to 90 °C, such as from 15 °C to 85 °C, for example from 20 °C to 80 °C, such as about 70 °C.

[0233] In some embodiments, in (vii-g) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

[0234] In some embodiments, the method further comprises either [1] (viii- g1) selectively deprotecting Compound (9b) in a mixture to form Compound (9c) 100Ref: P329WO , wherein PG is a protecting group,mpound (10) ; or [2] (viii-g2) seleo form Compound (9) and (ix-g2) depro.

[0235] (vi-h) reacting Compound (6with ethylene glycol chlorophosphate to form Compound (8b)101Ref: P329WO .

[0236] In ser comprises either [1] (vii- h1) converting Compound (8b) to Compound (9) and (viii-h1) depr; [2] (vii-h2) conve, wherein PG is a protecting group,b) in a mixture to form Compound (9c) , wherein PG is a protecting group,Ref: P329WO and (ix-h2) deprotecting Compound (9c) to form Compound (10) ; or [3] (vii-h3) con, wherein PG is a protecting group,) to form Compound (9)and (ix-h3) deprotecting Compound (9) in a mixture to form Compound (10) .

[0237] ing Compound (8b) with ammonia or an ammonium salt, or an ammonia surrogate.

[0238] In some embodiments, (vii-h1) comprises reacting Compound (8b) with ammonia or an ammonium halide, such as ammonium chloride or methanolic ammonia. 103Ref: P329WO

[0239] In some embodiments, (vii-h2) comprises reacting Compound (8b) with a protected amine or an ammonia derivative having a protecting group such as a benzyl or allyl protecting group, such as benzylamine, 4-methoxybenzylamine, or allylamine (e.g., PGNH2), followed by removal of the protecting group. In some embodiments, (vii-h3) comprises reacting Compound (8b) with a protected amine or an ammonia derivative having a protecting group such as a benzyl or allyl protecting group, such as benzylamine, 4-methoxybenzylamine, or allylamine (e.g., PGNH2), followed by removal of the protecting group.

[0240] In some embodiments, in (vii-h2) or (vii-h3), Compound (8b) is reacted with an amine selected from the group consisting of: a. benzylamine, to provide Compound (9b), wherein PG is benzyl; b. 4-methoxybenzylamine, to provide Compound (9b), wherein PG is 4- methoxybenzyl; and c. allylamine, to provide Compound (9b), wherein PG is allyl.

[0241] In some embodiments, at least 1.0 equivalent of the amine is added, such as from 1.0 to 5.0 equivalents, such as from 1.1 equivalents to 4.5 equivalents, for example from 1.5 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

[0242] In some embodiments, in (vii-h1), (vii-h2) or (vii-h3), a base is added to the solution, such as an amine base or a metal carbonate.

[0243] In some embodiments, the amine base is triethylamine.

[0244] In some embodiments, in (vii-h1), (vii-h2) or (vii-h3) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (8b), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

[0245] In some embodiments, in (vii-h1), (vii-h2) or (vii-h3) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent or a chlorinated solvent, for example wherein the polar aprotic solvent is selected from the group consisting of: THF, DCM, 2-MeTHF, DMSO, DMF, MeCN, diethyl ether, and a mixture thereof. 104Ref: P329WO

[0246] In some embodiments, in (vii-h1), (vii-h2) or (vii-h3) the temperature is from 0 °C to 120 °C, such as from 10 °C to 110 °C, for example from 20 °C to 100 °C.

[0247] In some embodiments, in (vii-h1), (vii-h2) or (vii-h3) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

[0248] In some embodiments, a method is provided for synthesizing Compound (10) , or a salt thereof, the method comp(vi-h) reacting Compound (6)with ethylene glycol chlorophosphate to form Compound (8b); r[1] (vii-h1) converting Compound (8b) to Compound (9) d105Ref: P329WO (viii-h1) deprotecting Compound (9) to form Compound (10); , or[2] (vii-h2’) reacting a compound HNPG2 with Compound (8b) to yield Compound , wherein each PG is independently(viii-h2’) selectively deprotecting Compound (9b’) in a mixture to form Compound (9c’), and(ix-h2) deprotecting Compound (9c’) to form Compound (10); or106Ref: P329WO [3] (vii-h3’) reacting a compound HNPG2 with Compound (8b) to yield Compound , wherein each PG is(viii-h3) selectively deprotecting Compound (9b’) to form Compound (9) and(ix-h3) deprotecting Compound (9) in a mixture to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

[0249] In some embodiments, in (vii-h2’) or (vii-h3’), Compound (8b) is reacted with dibenzylamine, to provide Compound (9b’), wherein each PG is benzyl.

[0250] In some embodiments, at least 1.0 equivalent of dibenzylamine is added, such as from 1.0 to 5.0 equivalents, such as from 1.1 equivalents to 4.5 equivalents, for example from 1.5 equivalents to 4.0 equivalents, for example about 1.1 equivalents. 107Ref: P329WO

[0251] In some embodiments, in (vii-h2’) or (vii-h3’), a base is added to the solution, such as an amine base or a metal carbonate.

[0252] In some embodiments, the amine base is triethylamine.

[0253] In some embodiments, in (vii-h2’) or (vii-h3’) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (8b), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

[0254] In some embodiments, in (vii-h2’) or (vii-h3’) the solution comprises a polar aprotic solvent, for example wherein the polar aprotic solvent is selected from the group consisting of: DMSO, and DMF, and a mixture thereof.

[0255] In some embodiments, in (vii-h2’) or (vii-h3’) the temperature is from 0 °C to 120 °C, such as from 10 °C to 110 °C, for example from 20 °C to 100 °C.

[0256] In some embodiments, in (vii-h2’) or (vii-h3’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

[0257] In some embodiments, a method for synthesizing Compound (10) , or a salt thereof, comprises(vi-f) phosphorylating the primary alcohol of Compound (6to form Compound (7);108Ref: P329WO (vii-f’) reacting a di-protected amino compound HOCH2CH2NPG2 with Compound (7) to yield Compound (9b’), wherein eachPG is independently a protecting group, sucan allyl or benzyl protecting group, and either [1] (viii-f1’) selectively deprotecting Compound (9b’) in a mixture to form Compound (9) andund (10); orn a mixture to form Compound (9c’)and (ix-f2) deprotecting Compound (9c’) to form Compound (10) 109Ref: P329WO , wherein R1is a C3-C30alkyl, C3-

[0258] In some embodiments, in (vii-f’), the di-protected amino compound HOCH2CH2NPG2 is N,N-dibenzyl-2-aminoethanol to provide Compound (9b’), wherein each PG is benzyl.

[0259] In some embodiments, in (vii-f’), a base is added to the solution, such as an amine base or a metal carbonate.

[0260] In some embodiments, the amine base is triethylamine.

[0261] In some embodiments, in (vii-f’) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (7), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

[0262] In some embodiments, in (vii-f’) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

[0263] In some embodiments, in (vii-f’) the temperature is from -10 °C to 80 °C, such as from 0 °C to 70 °C, for example from 10 °C to 60 °C.

[0264] In some embodiments, in (vii-f’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

[0265] In some embodiments, a method for synthesizing Compound (10) 110Ref: P329WO

[0266] , or a salt thereof, comprises(vi-f) phosphorylating the primary alcohol of Compound (6to form Compound (7);(vii-f’’) reacting a compound HOCH2CH2N+PG3 X- with Compound (7) to yield Compound (9b’’, wherein each PG is independently a protecting group, such as an allyl or benzyl group, and X- is a counter anion, such as Cl-, Br-, I-, OH-, or other suitable counter anion, and either [1] (viii-f1’’) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9) d(10); 111Ref: P329WO orin a mixture to form Compound (9c’’) dorm Compound (10) , wherein R1is a C3-C30alkyl, C3-

[0267] In some embodiments, a method for synthesizing Compound (10) , or a salt thereof, comprises112Ref: P329WO (vi-g) reacting Compound (6)with 2-bromoethyl phosphorodichloridate to form Compound (9a);(vii-g’) converting Compound (9a) to Compound (9b') by reacting a compound HNPG2 with Compound (9a) to yield Compound (9b’) , wherein each PG is independently ap; and either [1] (viii-g1) selectively deprotecting Compound (9b’) in a mixture to form Compound (9c’) andpound (10); 113Ref: P329WOor [2] (viii-g2) selectively deprotecting Compound (9b’) in a mixture to form Compound (9) d(10) ;(vii-gx) converting Compound (9a) to Compound (9), , wherein R1is a C3-C30alkyl, C3-

[0268] In some embodiments, (vii-gx) comprises reacting Compound 9a with ammonia, for example methanolic ammonia. 114Ref: P329WO

[0269] In some embodiments, in (vii-gx) no other base than ammonia is added.

[0270] In some embodiments, in (vii-gx) from 1.0 equivalents to 5.0 equivalents of a base is used with respect to Compound (9a), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents, for example wherein the base is an amine base, such as triethylamine.

[0271] In some embodiments, in (vii-gx) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

[0272] In some embodiments, in (vii-gx) the temperature is from 30 °C to 100 °C, such as from 35 °C to 90 °C, for example from 40 °C to 85 °C.

[0273] In some embodiments, in (vii-gx) the reaction is performed over the course of at least 30 minutes, such as at least 45 minutes, such as at least 1 hour, such as from 30 minutes to 120 hours.

[0274] In some embodiments, in (vii-gx) from 10 to 50 equivalents of ammonia is used, such as from 10 to 40 equivalents, for example 20 equivalents.In some embodiments, in (vii-g’), Compound 9a is reacted with a di-protected amine (PG2NH), optionally wherein the di-protected amine is dibenzylamine.

[0275] In some embodiments, in (vii-g’) from 1.0 to 5.0 equivalents of the protected amine is used, such as from 1.2 equivalents to 5.0 equivalents, for example from 1.5 equivalents to 4.0 equivalents, such as about 3.0 equivalents or such as about 4.0 equivalents.

[0276] In some embodiments, in (vii-g’) a base is used, such as an amine base.

[0277] In some embodiments, the amine base is triethylamine.

[0278] In some embodiments, in (vii-g’) Compound (9a) is reacted with the di-protected amine in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether. 115Ref: P329WO

[0279] In some embodiments, in (vii-g’) the temperature is from 10 °C to 90 °C, such as from 15 °C to 85 °C, for example from 20 °C to 80 °C, such as about 70 °C.

[0280] In some embodiments, in (vii-g’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

[0281] In some embodiments, a method for synthesizing Compound (10) dphosphorodichloridate to form Compound (9a);(vii-g’’) converting Compound (9a) to Compound (9b'’) by reacting a compound NPG3 with Compound (9a) to yield Compoun ,wherein each PG is independently a proup; and either 116Ref: P329WO [1] (viii-g1) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9c’’) dorm Compound (10); orb’’) in a mixture to form Compound (9) d(10) , wherein R1is a C3-C30alkyl, C3-

[0282] In some embodiments, in (vii-g’’), Compound 9a is reacted with a tri-protected amine (PG3N), optionally wherein the tri-protected amine is trimethylamine.

[0283] In some embodiments, in (vii-g’’) a metal oxide catalyst is added, such as wherein the metal oxide catalyst is silver oxide, for example wherein from 0.5 to 117Ref: P329WO 5.0 equivalents of the metal oxide catalyst is used, such as from 0.6 equivalents to 5.0 equivalents, for example from 0.7 equivalents to 4.0 equivalents, such as about 1.0 equivalents.

[0284] In some embodiments, in (vii-g’’) Compound (9a) is reacted with the tri-protected amine in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

[0285] In some embodiments, in (vii-g’’) the temperature is from 10 °C to 90 °C, such as from 15 °C to 85 °C, for example from 20 °C to 80 °C, such as about 70 °C.

[0286] In some embodiments, in (vii-g’’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

[0287] In some embodiments, a method for synthesizing Compound (10) sphosphorodichloridate to form Compound (9a);118Ref: P329WO (vii-g’’’) converting Compound (9a) to Compound (9b'’’) by reacting Compound (9a) with an ammonia surrogate, such as a metal azide, such as sodium azide, to yield Compoun d either [1] (viii-g1to form Compound (9c’’’)and (ix-g1) reducing Compound (9c’’’) to form Compound (10); oround (9) dd (10) 119Ref: P329WO , wherein R1is a C3-C30alkyl, C3-

[0288] In some embodiments, the metal azide in (vii-g’’’) is selected from the group consisting of: sodium azide, and potassium azide.

[0289] In some embodiments, in (vii-g’’’) from 1.0 to 10.0 equivalents of the metal azide is used, such as from 1.5 to 9.0 equivalents, such as from 2.0 to 8.0 equivalents, such as from 3.0 to 7.0 equivalents, for example about from 4.0 to 6.0 equivalents, for example about 5.0 equivalents.

[0290] In some embodiments, in (vii-g’’’) Compound (9a) is reacted with the metal azide in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

[0291] In some embodiments, in (vii-g’’’) the temperature is from 0 °C to 100 °C, such as from 15 °C to 90 °C, for example from 20 °C to 80 °C.

[0292] In some embodiments, in (vii-g’’’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

[0293] In some embodiments, the ammonia surrogate in (vii-h2’’’) or (vii- h3’’’) is a metal azide, such as a metal azide selected from the group consisting of: sodium azide, and potassium azide.

[0294] In some embodiments, in (vii-h2’’’) or (vii-h3’’’) from 1.0 to 10.0 equivalents of the metal azide is used, such as from 1.5 to 9.0 equivalents, such as from 2.0 to 8.0 equivalents, such as from 3.0 to 7.0 equivalents, for example about from 4.0 to 6.0 equivalents, for example about 5.0 equivalents.

[0295] In some embodiments, in (vii-h2’’’) or (vii-h3’’’) Compound (8b) is reacted with the metal azide in a solvent which is: a polar aprotic solvent, a polar protic 120Ref: P329WO solvent, or a combination thereof, for example wherein the solvent is selected from the group consisting of: H2O, THF, 2-MeTHF, acetone, and a mixture thereof; for example wherein the solvent comprises H2O and acetone in a from 1:5 to 1:2 mixture, such as in a 1:3 mixture.

[0296] In some embodiments, in (vii-h2’’’) or (vii-h3’’’) the temperature is from 0 °C to 100 °C, such as from 15 °C to 90 °C, for example from 20 °C to 80 °C.

[0297] In some embodiments, in (vii-h2’’’) or (vii-h3’’’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

[0298] In some embodiments of the methods disclosed above, Compound (10) is selected from d

[0299] s for synthesizing bioactive in vivo precursors to plasmalogen compounds. In some embodiments, the disclosure provides methods for synthesizing LPC(O)s. In some embodiments, the disclosure provides a method for synthesizing Compound (12) or a salt thereof 121Ref: P329WO .

[0300] vi-a) reacting Compound (6) with 2-bromoethyl phosphorodichloridate to form Compound (9a) .

[0301] prises (vii-a) reacting Compound (9a) in a solution comprising a metal oxide and trimethylamine to form Compound (11) .

[0302] ) is silver oxide.

[0303] In some embodiments, the method further comprises (viii-a) deprotecting Compound (11) to form Compound (12) .122Ref: P329WO

[0304] In some embodiments, (viii-a) comprises reacting Compound (11) with at least one catalyst. In some embodiments, (viii-a) comprises reacting Compound (11) with at least one catalyst and under an inert atmosphere.

[0305] In some embodiments, in (viii-a), the at least one catalyst may be selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. In some embodiments, a catalyst derived from any of the foregoing or generated in situ from any of the foregoing by combining any of the foregoing with a ligand, such as a phosphine ligand, such as triphenylphosphine, tri-tert-butylphosphine, or 1,1′-ferrocenediyl-bis(diphenylphosphine). In some embodiments, the catalyst is tetrakis(triphenylphosphine)palladium(0). However, one of ordinary skill in the art would also understand that alternate deprotection catalysts may be suitable for the removal of an allyl protecting group, as described elsewhere here.

[0306] In some embodiments, the method comprises (vi-b) phosphorylating the primary alcohol of Compound (6) to form Compound (7).

[0307] In some embodiments, (vi-b) comprises reacting Compound (6) with a phosphoryl halide to form Compound (7). In some embodiments, (vi-b) comprises reacting Compound (6) with a phosphoryl halide at a temperature of about 0°C under an inert atmosphere. In some embodiments, the phosphoryl halide is phosphoryl trichloride.

[0308] In some embodiments, (vi-b) further comprises dissolving Compound (6) in a solution comprising triethylamine and diethyl ether.

[0309] In some embodiments, (vi-b) comprises adding a solution of Compound (6) to a solution comprising phosphoryl halide dropwise. 123Ref: P329WO

[0310] In some embodiments, the method further comprises (vii-b) reacting Compound (7) with ethanolamine in solution to form Compound (8) .

[0311] In smprises dissolving Compound (7) in a solution comprising diethyl ether and tetrahydrofuran. In some embodiments, the solution further comprises ethanolamine and triethylamine.

[0312] In some embodiments, (vii-b) further comprises heating the solution. In some embodiments, (vii-b) comprises heating the solution under an inert atmosphere. In some embodiments, (vii-b) comprises heating the solution at a temperature range of about 30°C to 60°C under an inert atmosphere. In some embodiments, (vii-b) comprises heating the solution at a temperature range of about 25°C to 70°C, about 30°C to 60°C, or about 40°C to 50°C. In some embodiments, (vii- b) comprises heating the solution at a temperature of about 50°C with an internal temperature of 40°C under an inert atmosphere.

[0313] In some embodiments, the method further comprises (viii-b) reacting Compound (8) in a mixture comprising an acid to form Compound (9).

[0314] In some embodiments, the acid in (viii-b) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, a carboxylic acid, a sulfonic acid, and an alkali metal dihydrogen phosphate. In some embodiments, the acid in (viii-b) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate. In some embodiments, the acid in (viii-b) is hydrochloric acid. 124Ref: P329WO

[0315] In some embodiments, in (viii-b), the mixture comprises diethyl ether and tetrahydrofuran. Synthesis of Compound (12)

[0316] In some embodiments, the method further comprises (ix-b) methylating Compound (9) to form Compound (11) .hod is provided for producing Compound (12), wherein the method comprises deprotecting Compound (11) using a method as defined herien, for example by (x-b), to form Compound (12).

[0318] In some embodiments, methylating Compound 9 comprises reacting Compound (9) with methyl bromide, methyl iodide, or dimethyl sulfate.

[0319] In some embodiments, the method further comprises (x-b) deprotecting Compound (11) to form Compound (12) .comprises (1-a) reacting R1-OH with (2R)-(-)-glycidyl tosylate wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group (Compound (18)) to form Compound (19) ).125Ref: P329WO

[0321] In some embodiments, the method further comprises (1-b) reacting Compound (19) with a benzylating reagent in solution, the benzylating reagent reacting with the secondary alcohol of Compound (19) to form Compound (20) ), wherein Ar is selected from phyl.

[0322] In some embodiments, the method further comprises (1-c) reacting Compound (20) with a metal acetate to form Compound (21) ).

[0323] As usedsalt of a metal cation and the acetate (CH3CO2-) anion. In some embodiments, the metal acetate is caesium acetate.

[0324] In some embodiments, the method further comprises (1-d) hydrolyzing Compound (21) to form Compound (22) ).

[0325] In some ed further comprises (1-e) reacting Compound (22) with ethylene chlorophosphate to form Compound (23) ).

[0326] In somerther comprises (1-f) reacting Compound (23) in a solution comprising trimethylamine to form Compound (24) 126Ref: P329WO ).

[0327] Icomprises (1-g) deprotecting Compound (24) to form Compound (12) .

[0328] d above, Compound (12) is selected from d

[0329] In some embodiments, the disclosure provides a method for synthesizing Compound (25) or a salt thereof 127Ref: P329WO .

[0330] (2-a) providing Compound (26), wherein each of R1and R3is independently selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, .

[0331] In some ed further comprises (2-b) phosphorylating the primary alcohol of Compound (26) to form Compound (27) .

[0332] In soher comprises (2-c) reacting Compound (27) with ethanolamine in solution to form Compound (28) .

[0333] In soer comprises (2-d) reacting Compound (28) in a mixture comprising acid to form Compound (25) .Ref: P329WO

[0334] In some embodiments, a deprotection reaction removes the R3group from Compound (25), to form Compound 10 .

[0335] er reacted to form Compound 12 in a method comprising (3-a) methylating Compound (25) to form Compound (29) .

[0336] prises (3-b) deprotecting Compound (25) to form Compound (12) .

[0337] d above, R1is a C16- C18 alkyl, C16-C18 alkenyl, or C16-C18 acyl group. In some embodiments, R1is a C16 alkyl group. In some embodiments, R1is a C18 alkyl group. In some embodiments, R1is a C18 alkenyl group.

[0338] In some embodiments, R1is a C10-C24 alkyl, such as C15-C20 alkyl, for example a C16-C18 alkyl, C10-C24 alkenyl, such as a C15-C20 alkenyl, for example a C16-C18 alkenyl, or a C10-C24 acyl, such as a C15-C20 acyl, for example a C16-C18 acyl group. 129Ref: P329WO

[0339] In some embodiments of the methods disclosed above, R3is an allyl group. Strategies 1-3: Penultimate and Final Steps

[0340] The following section describes specific conditions for protecting group removal, azide to amine conversions, including specifically the penultimate or final steps of Strategies 1, 2, and 3 of Example 10. These conditions can be combined with any earlier steps and reaction conditions of the syntheses disclosed herein.

[0341] In some embodiments, in particular for the methods of Example 10, Strategy 1, N-protecting groups may be removed as set out below:

[0342] BOC: In some embodiments, methods of the present disclosure for removal of a BOC-group comprise treatment with aqueous HCl in ethyl acetate, in-situ generated HCl(g) in isopropyl acetate (from isopropanol and acetyl chloride), or BF₃·Et₂O in dichloromethane in the presence of 4Å molecular sieves.

[0343] Benzyl: In some embodiments, methods of the present disclosure for removal of benzyl comprise hydrogenation using H₂ over Pd / C (and variants) or oxidation with ceric ammonium nitrate in acetonitrile and water.

[0344] 4-MeO benzyl: In some embodiments, methods of the present disclosure for removal of 4-MeO benzyl comprise hydrogenation using H₂ over Pd / C (and variants), oxidation with ceric ammonium nitrate in acetonitrile and water, DDQ in dichloromethane and water, or trifluoroacetic acid in dichloromethane.

[0345] Allyl: In some embodiments, methods of the present disclosure for removal of allyl comprise Pd(PPh₃)₄ with dimethyl barbituric acid in THF; or BX₃-based N-deprotection.

[0346] Trityl: In some embodiments, methods of the present disclosure for removal of trityl comprise treatment with HCl in acetone, hydrogenation using H₂ over Pd / C, or Yb(OTf)₃ in THF / water.

[0347] Dibenzyl / tribenzyl: In some embodiments, methods of the present disclosure for removal of dibenzyl / tribenzyl comprise hydrogenation using H₂ over Pd / C (and variants) or oxidation with ceric ammonium nitrate in acetonitrile and water. 130Ref: P329WO

[0348] Diallyl: In some embodiments, methods of the present disclosure for removal of diallyl comprise Pd(PPh₃)₄ with dimethyl barbituric acid; and BX₃-based N-deprotection.

[0349] In some embodiments, in particular for the methods of Example 10, Strategy 1, O-protecting groups may be removed as set out below:

[0350] Allyl: In some embodiments, methods of the present disclosure for removal of allyl of the present disclosure for removal of an allyl protecting group comprise treatment with tetrakis(triphenylphosphine)palladium(0) [Pd(PPh₃)₄] and dimethyl barbituric acid in THF, as well as BF₃·Me₂S in dichloromethane. Other acidic ether cleavage conditions, such as treatment with mineral acids, may also be employed.

[0351] In some embodiments, in particular for the methods of Example 10, Strategy 2, N-protecting groups may be removed as set out below:

[0352] Benzyl: In some embodiments, methods of the present disclosure for removal of a benzyl protecting group comprise hydrogenation using H₂ over Pd / C (and variants) or oxidation with ceric ammonium nitrate in acetonitrile and water.

[0353] 4-MeO benzyl: In some embodiments, methods comprise hydrogenation using H₂ over Pd / C (and variants), ceric ammonium nitrate in acetonitrile and water, DDQ in dichloromethane and water, or trifluoroacetic acid in dichloromethane.

[0354] Allyl: In some embodiments, methods comprise Pd(PPh₃)₄ with dimethyl barbituric acid in THF and related variants.

[0355] Trityl: In some embodiments, methods comprise HCl in acetone, hydrogenation using H₂ over Pd / C, or Yb(OTf)₃ in THF and water.

[0356] 4-MeO trityl: In some embodiments, methods comprise acid hydrolysis using trichloroacetic acid (CCl₃CO₂H) or dichloroacetic acid (CHCl₂CO₂H) with anisole in dichloromethane.

[0357] Dibenzyl: In some embodiments, methods for dibenzyl group removal are assumed to follow those described for N-benzyl deprotection.

[0358] Diallyl: In some embodiments, methods comprise Pd(PPh₃)₄ with dimethyl barbituric acid in THF and related variants. 131Ref: P329WO

[0359] Tribenzyl: In some embodiments, methods for tribenzyl group removal are assumed to follow those described for N-benzyl deprotection.

[0360] Triallyl: In some embodiments, methods comprise Pd(PPh₃)₄ with dimethyl barbituric acid in THF and related variants.

[0361] In some embodiments, in particular for the methods of Example 10, Strategy 2, methods for conversion of azide to amine with an O-allyl as part of the molecule, not to be converted, are set out below:

[0362] H₂ with PtO₂, Pd, or Rh catalysts: In some embodiments, methods for azide to amine conversion comprise hydrogenation using H₂ in the presence of platinum dioxide (PtO₂), palladium (Pd), or rhodium (Rh) catalysts and their variants.

[0363] BF₃·OEt₂ with NaI or BF₃·Me₂S: In some embodiments, methods for azide to amine conversion comprise treatment with BF₃·OEt₂ and about 0.5 equivalents of sodium iodide in acetonitrile, or treatment with BF₃·Me₂S under analogous conditions.

[0364] Triphenylphosphine and water (Staudinger reaction): In some embodiments, methods for azide to amine conversion comprise treatment with triphenylphosphine (PPh₃) and water, or variants thereof, to achieve reduction via the Staudinger reaction.

[0365] In some embodiments, in particular for the methods of Example 10, Strategy 3, methods for conversion of azide to amine with an O-allyl as part of the molecule, not to be converted, are set out below:

[0366] H₂ with PtO₂, Pd, or Rh catalysts: In some embodiments, methods for azide to amine conversion comprise hydrogenation using H₂ in the presence of platinum dioxide (PtO₂), palladium (Pd), or rhodium (Rh) catalysts and their variants.

[0367] BF₃·OEt₂ with NaI or BF₃·Me₂S: In some embodiments, methods for azide to amine conversion comprise treatment with BF₃·OEt₂ and about 0.5 equivalents of sodium iodide in acetonitrile, or treatment with BF₃·Me₂S in dichloromethane.

[0368] Triphenylphosphine and water (Staudinger reaction): In some embodiments, methods for azide to amine conversion comprise treatment with 132Ref: P329WO triphenylphosphine (PPh₃) and water, or variants thereof, to achieve reduction via the Staudinger reaction.

[0369] In some embodiments, the methods disclosed herein further comprises a step of a global deprotection comprising simultaneous removal of O-allyl and N-allyl protection (BOC, trityl and allyl) as a final step. Strategy Exemplary protecting groups Structure

[0370] In some embodiments, global deprotection of the reactant of strategy 1 outlined above under “structure” or an analogous compound with different protecting groups comprises treatment of the reactant with acid, such as HCl, such as 4M HCl in ethyl acetate, for example at between 0 °C and 75 °C, for example about 25 °C.

[0371] In some embodiments, global deprotection of the reactant of strategy 2 outlined above under “structure” or an analogous compound with different protecting groups comprises treatment of the reactant with acid, such as HCl, such as concentrated HCl in acetone, for example at between 0 °C and 75 °C, for example about 25 °C.

[0372] In some embodiments, global deprotection of the reactant of strategy 2 outlined above under “structure” or an analogous compound with different protecting groups comprises treatment of the reactant with a palladium catalyst, such as 133Ref: P329WO a palladium catalyst disclosed herein, for example , such as HCl, such as concentrated HCl in acetone, for example at between 0 °C and 75 °C, for example about 25 °C. Compounds

[0373] In some embodiments, the disclosure provides a compound selected from Compound (6),Comp,134Ref: P329WO Compoun ,Compoun ,Compound (10 ,Compound (11),Compoun ,135Ref: P329WO and a salt of any of the foregoing, wherein each of R1and R3is independently selected from a C16-C18 alkyl group, a C16-C18 alkenyl group, and a C16-C18 acyl group. In some embodiments, the disclosure provides a composition comprising the same.

[0374] In some embodiments, the present disclosure provides a compound selected from the group consisting of:

[0375] (R)-4-((hexadecyloxy)methyl)-2,2-dimethyl-1,3-dioxolane ;ioxolane e;;136Ref: P329WO

[0381] 2-((R)-2-(allyloxy)-3-(hexadecyloxy)propoxy)-1,3,2- oxazaphospholidine 2-oxide ;oxazaphospholidine 2-oxide ;1,3,2oxazaphospholidine 2-oxide ;phosphate ;137Ref: P329WO

[0385] (R)-2-(allyloxy)-3-(octadecyloxy)propyl (2-ammonioethyl) phosphate ;phosphate ;yl) phosphate ;hyl) phosphate ;(trimethylammonio)ethyl phosphate 138Ref: P329WO

[0390] In some embodiments of the compounds and compositions disclosed above, R1is a C16 alkyl group. In some embodiments, R1is a C18 alkyl group. In some embodiments, R1is a C18 alkenyl group.

[0391] In some embodiments, the compounds disclosed herein are not marine-sourced (i.e., the compounds disclosed herein are not obtained from marine sources). In some embodiments, the compounds disclosed herein are not obtained 139Ref: P329WO from shark liver oil. In some embodiments, the compounds disclosed herein are not obtained from krill. Further Transformations

[0392] Disclosed herein are methods of using the chemical precursors to bioactive compounds disclosed herein to prepare plasmalogen compounds.

[0393] In some embodiments, the chemical precursors to bioactive compounds disclosed herein are subjected to one or more of the following transformations: (A) dehydrogenation of an R1group (at the P1position), and (B) acylation of the secondary alcohol to incorporate an acyl group.

[0394] In some embodiments, any of the above methods disclosed herein may be combined with any of the transformations outlined in this section.

[0395] In some embodiments, any method step disclosed herein may be performed in isolation and may form an independent embodiment.

[0396] Transformation (A) is illustrated in the following Scheme A, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group; R4is selected from CH2-CH2-N-Me3+and CH2-CH2-NH2; and R5is selected from a C1-C28 alkyl group, a C1-C28 alkenyl group, and a C1-C28 acyl group. 1.

[0397] Transformation (B) is illustrated in the following Scheme B, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group; R4is selected from CH2-CH2-N-Me3+and CH2-CH2-NH2; and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group. 140Ref: P329WO

[0398] In some embodiments of Transformation (A) and / or Transformation (B), R1is selected from a C14-C24 alkyl group, a C14-C24 alkenyl group, and a C14-C24 acyl group. In some embodiments, R1is selected from a C16-C18 alkyl group, a C16-C18 alkenyl group, and a C16-C18 acyl group. In some embodiments, R1is a C16 alkyl group. In some embodiments, R1is a C18 alkyl group. In some embodiments, R1is a C18 alkenyl group.

[0399] In some embodiments of Transformation (A) and / or Transformation (B), R5is selected from a C12-C22 alkyl group, a C12-C22 alkenyl group, and a C12-C22 acyl group. In some embodiments, R5is selected from a C14-C16 alkyl group, a C14-C16 alkenyl group, and a C14-C16 acyl group. In some embodiments, R5is a C14 alkyl group. In some embodiments, R1is a C16 alkyl group. In some embodiments, R1is a C16 alkenyl group.

[0400] In some embodiments of Transformation (A) and / or Transformation (B), R6is selected from a C14-C24 alkyl group and a C14-C24 alkenyl group. In some embodiments, R6is selected from a C16-C18 alkyl group and a C16-C18 alkenyl group.

[0401] In some embodiments, the chemical precursors to bioactive compounds of the disclosure are subjected to both transformation (A) and transformation (B) to yield a plasmalogen structure.

[0402] In some embodiments, transformation (A) converts the chemical precursor to bioactive compounds to their corresponding vinyl ethers.

[0403] In some embodiments, transformation (A) is achieved enzymatically, such as by using a desaturase enzyme. In some embodiments, the desaturase enzyme is TMEM189. See, e.g., Werner ER, et al., “The TMEM189 Gene 141Ref: P329WO Encodes Plasmanylethanolamine Desaturase which Introduces the Characteristic Vinyl Ether Double Bond into Plasmalogens,” PNAS, 117(14), pp.7792-98 (2020); S. Padmanabhan et al., “Plasmalogens and Photooxidative Stress Signaling in Myxobacteria, and How it Unmasked CarF / TMEM189 as the Δ1′-Desaturase PEDS1 for Human Plasmalogen Biosynthesis,” Front. Cell Dev. Biol., 10 (2020). However, one of ordinary skill in the art would also understand that alternative enzymes, such as alternative desaturase enzymes, may be suitable for the dehydrogenation of R1.

[0404] In some embodiments, transformation (B) is achieved using an esterification reaction. In some embodiments, the esterification reaction of transformation (B) uses an enzyme, such as an enzyme without phospholipase activity. In some embodiments, transformation (B) is accomplished via a P2-specific esterification reactions using an enzyme without phospholipase activity and in a mixture with low water activity present. In some embodiments, transformation (B) is carried out chemically by reacting a carboxylic acid, such as a fatty acid, and a compound disclosed herein having a secondary alcohol at the P2 position in the presence of a coupling reagent. Coupling reagents suitable for the transformation are known to persons having ordinary skill in the art and include, for example, dicyclohexyl carbodiimide (DCC). In some embodiments, transformation (B) is carried out chemically by converting a carboxylic acid, such as a fatty acid, to an acyl halide and reacting the acyl halide with a compound disclosed herein having a secondary alcohol at the P2 position.

[0405] Regarding transformation (A), without wishing to be bound by theory, a substitution at R1wherein there is a vinyl ether form of a C16 or C18 substituted moiety is an intermediate thought to ultimately lead to the production of plasmalogens (PE-P or PC-P) in mammals. Regarding transformation (B), without wishing to be bound by theory, PC(O) and PE(O) compounds, and particularly those substituted with DHA and EPA at P2, are also thought to be biologically active or precursors for plasmalogens.

[0406] In some embodiments, the chemical precursor to a plasmalogen compound subjected to transformations (A) and / or (B) is selected from: 142Ref: P329WO ap .

[0407] In some embodiments, Compound (1subjected to Transformation (A) and / or Transformation (B) as shown in the following Scheme C, wherein R1is defined as disclosed herein, for example selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group; R5is selected from a C1-C28 alkyl group, a C1-C28 alkenyl group, and a C1-C28 acyl group; and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group. 143Ref: P329WO0).

[0408] In some embodiments, Compound (12) is subjected to Transformation (A) and / or Transformation (B) as shown in the following Scheme D wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group; R5is selected from a C1-C28 alkyl group, a C1-C28 alkenyl group, and a C1- C28 acyl group; and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group. In some embodiments, R1is selected from a C16 alkyl group, a C18 alkyl group, and a C18 alkenyl group. 144Ref: P329WO).

[0409] In some embodiments, Compound (9) is subjected to Transformation (A) as shown in the following Scheme E wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group and R5is selected from a C1-C28 alkyl group, a C1-C28 alkenyl group, and a C1-C28 acyl group. In some embodiments, R1is selected from a C16 alkyl group, a C18 alkyl group, and a C18 alkenyl group.

[0410] In some embodiments, for Compound (9), R1 is selected from the group consisting of: hexadecanyl; octadecanyl, and (Z)-octadec-9-en-1-yl.

[0411] In some embodiments, Compound (11) is subjected to Transformation (A) as shown in the following Scheme F wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group and R5is selected from a C1-C28 alkyl group, a C1-C28 alkenyl group, and a C1-C28 acyl group. In some 145Ref: P329WO embodiments, R1is selected from a C16 alkyl group, a C18 alkyl group, and a C18 alkenyl group. O O (A) e3

[0412] In some embodiments, applying Transformation (A) and (B) to Compound (12) yields the following plasmalogen compound: .EXAMPLES

[0413] In the context of the present disclosure, the following abbreviations are used. 2-MeTHF 2-Methyltetrahydrofuran CAD Charged Aerosol Detector CV Column volumes DCM Dichloromethane DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide EA Ethyl acetate ELSD Evaporative Light-Scattering Detector HPLC High Performance Liquid Chromatography HSE Health, safety and environment KHMDS Potassium bis(trimethylsilyl)amide IPA Isopropanol IPAc Isopropyl acetate 146Ref: P329WO IY Intermediate Yield LC-MS Liquid Chromatography-Tandem Mass Spectrometry LiHMDS Lithium bis(trimethylsilyl)amide MeCN or ACN Acetonitrile MTBE Methyl tert-butyl ether NaHMDS Sodium bis(trimethylsilyl)amide NMR Nuclear Magnetic Resonance PMI Potentially mutagenic impurity TBAB Tetrabutylammonium bromide TBSCl tert-Butyldimethylsilyl chloride t-BuOK Potassium tert-butoxide TEA Triethylamine

[0414] Unless stated otherwise throughout the present examples, R1 is C16H33 (hexadecanyl) C18H37 (octadecanyl), or C18H35 ((Z)-octadec-9-en-1-yl). Example 1: Synthetic Route 1 – Synthesis of Compound (6)

[0415] Synthesis of Compound (6). Each of the reactions and numbered intermediates described below refer to the corresponding reactions and intermediates in Scheme II. 147Ref: P329WO HO C18H35OH Mesyl chloride Et3N, DCM C16H33BrC18H35OMsCompound 1 nd 1 TB (i) Compou AB TBABNaOH, H2OCompound 1 NaOH, H2OTBAB NaOH, H2OR1O O (R1= C16H33, C18H37.or O 2 C18H35) HCl, H2O (ii) THF R1O OH 3 OH TBSCl imidazole Pyridin DMF (iii) 4 Ally Na 5 6Scheme II. Exemplary Synthesis of Compound 6. 148Ref: P329WO

[0416] Reaction (i) – Formation of Compound (2): A 500 mL three-neck flask was fitted with a stir bar, waterless condenser, and a thermometer. The flask was charged with a freshly-prepared aqueous sodium hydroxide solution (19 M, 142 mL, 37.5 equiv.) followed by Compound (1) (9.5 g, 71.9 mmol, 1.0 equiv.) [Note 1]. The resulting solution was heated to 80 °C and once at temperature, 1-bromohexadecane (R1= C16H33) (85 g, 278 mmol, 3.9 equiv.) was added in a single portion, followed by tetrabutylammonium bromide (4.63 g, 14.4 mmol, 0.2 equiv.) [Note 2]; [Note 3]. The resulting clear solution was stirred vigorously for 3 days at 80 °C. The reaction mixture was cooled to ambient temperature then diluted with dichloromethane (250 mL) and transferred to a separating funnel [Note 4]. The layers were separated and the aqueous layer extracted with dichloromethane (2 x 250 mL). The organic layers were washed with water (200 mL) and brine (200 mL) then dried over sodium sulfate, filtered, and concentrated to give a colorless wax (74 g) [Note 5]. The crude product was dissolved in dichloromethane (200 mL) and silica gel added. The mixture was concentrated to give a homogeneous powder which was dry-loaded and purified by normal-phase chromatography: Biotage Isolera LS, 800 g SiliCycle cartridge; eluent 100% heptane for 3 CV then 5% ethyl acetate in heptane for 5 CV then 7% ethyl acetate in heptane for 2 CV. Product fractions were concentrated to give 25.4 g of a colorless wax which was further purified by normal-phase chromatography: Biotage Isolera LS, 330 g SiliCycle cartridge; eluent 0-5% ethyl acetate in heptane over 10 CV. Product fractions were combined and concentrated to give Compound (2) (R1= C16H33) (17.4 g, 100% estimated purity by NMR, 68% yield) as a colorless wax.1H NMR (400 MHz, CDCl3, ppm): d 4.27 (quint, J = 6.0 Hz, 1 H), 4.07 (dd, J = 8.3, 6.4 Hz, 1 H), 3.74 (dd, J = 8.3, 6.4 Hz, 1H), 3.55-3.40 (m, 4 H), 1.62-1.56 (m, 2 H), 1.43 (s, 3 H), 1.37 (s, 3 H), 1.26 (s, 26 H), 0.93-0.86 (m, 3 H).13C NMR (100 MHz, CDCl3): δ 109.3, 74.7, 71.8, 71.7, 66.9, 31.9, 29.7-29.6 (8 C), 29.5, 29.4, 29.3, 26.7, 26.0, 25.3, 22.6, 14.1.

[0417] Notes on Reaction (i): [1] 142 mL of 19 M NaOH requires 108 g of NaOH powder. The dissolution process can be extremely exothermic if not done slowly and cautiously. 149Ref: P329WO [2] A significant decrease in reaction temperature was observed on addition of the 1- bromohexadecane as there was a large amount of material and it was at room temperature. No subsequent exotherm was noted. [3] The 1-bromohexadecane is a low-melting solid. To aid transfer to the reaction mixture it was gently melted with a heat gun prior to addition. [4] On cooling to room temperature a thick gel formed due to both the product and by- products being waxy solids at room temperature. [5] This material contained significant amounts of 1-hexadecanol and hexadec-1-ene.

[0418] An analogous transformation was also carried out using 1- bromooctadecane (R1= C18H37) and Compound (1) to yield Compound (2) wherein R1= C18H37.

[0419] The conversion of (Z)-octadec-9-en-1-ol teflask was charged with a stir bar and dichloromethane (300 mL), followed by C18H35OH (42.0 g, 141 mmol, 1.0 equiv.) and triethylamine (21.6 mL, 155 mmol, 1.1 equiv.) [Note 1]. The flask was cooled to 0 °C. Then methanesulfonyl chloride (12.0 mL, 155 mmol, 1.1 equiv.) was added dropwise over 5 mins. The flask was stirred at 0 °C for a further 5 minutes, then warmed to ambient temperature and stirred for 3 hours. The reaction mixture was quenched by addition of saturated aqueous sodium bicarbonate solution (100 mL) and the layers separated. The organic layer was washed with saturated aqueous ammonium chloride solution (100 mL) then brine (200 mL). The resulting product was dried over magnesium sulfate, filtered, and concentrated to give C18H35OMs (52.0 g, crude) as a light-yellow oil [Note 2].1H NMR (400 MHz, CDCl3, ppm): δ 5.42 - 5.32 (m, 2 H) 4.23 (t, J=6.6 Hz, 2 H) 3.01 (s, 3 H) 1.95 - 2.09 (m, 4 H) 150Ref: P329WO 1.70 - 1.82 (m, 2 H) 1.25 - 1.37 (m, 22 H) 0.86 - 0.93 (m, 3 H). Then, an analogous transformation as described above for converting Compound (1) to Compound (2) was carried out using C18H35OMs and Compound (1) to yield Compound (2) wherein R1= C18H35.

[0420] Notes on the conversion of C18H35OH to C18H35OMs: [1] C18H35OH was used as supplied (technical grade [85% purity]). [2] Q-NMR data was not available for this experiment, however, the same step gave a yield of 94% (corrected for Q-NMR purity) for a different experiment (30 g scale).

[0421] Reaction (ii) – Formation of Compound (3): A 3-necked flask was fitted with a thermometer, stir bar, and waterless condenser. The flask was charged with Compound (2) (R1= C16H33) (17.4 g, 48.8 mmol, 1.0 equiv.), tetrahydrofuran (135 mL), and aqueous hydrochloric acid solution (0.5 M, 59 mL, 0.6 equiv.). The resulting mixture was heated to 65 °C for 6.5 h then cooled to ambient temperature and concentrated in vacuo to remove excess tetrahydrofuran. The resulting residue was neutralized with saturated aqueous sodium bicarbonate solution (50 mL). The aqueous layer was extracted with dichloromethane (3 x 200 mL), and the combined organic layers were washed with brine (200 mL) then dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain Compound (3) (R1= C16H33) (14.7 g, crude) as a colorless solid [Note 1].1H NMR (400 MHz, CDCl3, ppm): d 3.91-3.84 (m, 1 H), 3.74-3.70 (m, 1H), 3.69-3.64 (m, 1 H), 3.57-3.44 (m, 4 H), 2.57 (br s, 1 H), 2.18 (br s, 1 H), 1.62-1.58 (m, 2 H), 1.37-1.24 (m, 26 H), 0.91-0.86 (m, 3 H).

[0422] Notes on reaction (ii): [1] Crude compound contains residual tetrahydrofuran, dichloromethane, and water.

[0423] Analogous transformations were also successfully carried out using Compound (2) to yield Compound (3) wherein R1= C18H37 and R1= C18H35.

[0424] Reaction (iii) – Formation of Compound (4): Imidazole (14.1 g, 207 mmol, 4.5 equiv.) was added to a stirred solution of Compound (3) (R1= C16H33) (14.7 g, 46.1 mmol, 1.0 equiv.) in a mixture of pyridine (105 mL), N,N-dimethylformamide (25 mL), and dichloromethane (25 mL). The mixture was then cooled to 0 °C. A solution of tert-butyldimethylsilyl chloride (7.64 g, 50.7 mmol, 1.1 equiv.) in pyridine (40 mL) was added dropwise and the reaction mixture stirred at 0 °C for 2 h, then warmed to ambient 151Ref: P329WO temperature and stirred for 18 h. Water (30 mL) was added and the resultant solution concentrated under reduced pressure to give Compound (4) (R1= C16H33) (19.1 g, crude) [Note 1].1H NMR (400 MHz, CDCl3, ppm): d 3.82 (sextet, J = 5.2 Hz, 1H), 3.70- 3.61 (m, 2 H), 3.50-3.40 (m, 4 H), 2.46 (d, J = 4.9 Hz, 1 H), 1.62 ‐ 1.53 (m, 2 H), 1.37- 1.26 (m, 26 H), 0.93-0.88 (m, 12 H), 0.11-0.06 (m, 6 H).

[0425] Notes on reaction (iii): [1] If needed toluene could be added to the residue then concentrated under reduced pressure to azeotrope any remaining solvents.

[0426] Analogous transformations were also successfully carried out using Compound (3) to yield Compound (4) wherein R1= C18H37 and R1= C18H35.

[0427] Reaction (iv) – Formation of Compound (5): A three-necked flask was charged with a stirrer bar and a solution of Compound (4) (R1= C16H33) (19.1 g, 43.9 mmol, 1.0 equiv.) in dry tetrahydrofuran (300 mL), then degassed with argon for 10 mins. Sodium hydride (60% dispersion in mineral oil, 3.51 g, 87.9 mmol, 2.0 equiv.) was added and the mixture stirred for 10 mins. The mixture was cooled to 0 °C and a solution of allyl bromide (10.6 g, 87.9 mmol, 2.0 equiv.) in dry tetrahydrofuran (40 mL) was added dropwise over 30 mins. The solution was stirred at 0 °C for 10 mins then warmed to ambient temperature and stirred for 16 h. The solution was cooled to 0 °C and water (20 mL) was added dropwise over 3 mins. The mixture was warmed to ambient temperature and stirred for 5 mins. The mixture was concentrated under reduced pressure and the residue partitioned between diethyl ether (150 mL) and water (150 mL). The layers were separated, and the aqueous layer extracted with diethyl ether (2 x 100 mL). The organic layers were combined and washed with brine (200 mL) and dried over sodium sulfate then filtered and concentrated under reduced pressure to give Compound (5) (R1= C16H33) (24.2 g, crude) as a dark-brown oil.1H NMR (400 MHz, CDCl3, ppm): d 5.99 - 5.86 (m, 1 H), 5.34 - 5.25 (m, 1 H), 5.19 - 5.12 (m, 1 H), 4.19 - 4.13 (m, 2 H), 3.71 - 3.63 (m, 2H), 3.59 - 3.51 (m, 2 H), 3.50 - 3.40 (m, 3 H), 1.62 - 1.56 (m, 2 H), 1.35 - 1.22 (m, 26 H), 0.93 - 0.85 (m, 12 H), 0.12 - 0.03 (m, 6 H).

[0428] Analogous transformations were also successfully carried out using Compound (4) to yield Compound (5) wherein R1= C18H37 and R1= C18H35. 152Ref: P329WO

[0429] Reaction (v) – Formation of Compound (6): A solution of tetrabutylammonium fluoride (1.0 M in tetrahydrofuran, 48.3 mL, 1.1 equiv.) was added to a stirred solution of Compound (5) (R1= C16H33) (20.90 g, 43.94 mmol, 1.0 equiv.) in tetrahydrofuran (200 mL) and the mixture was stirred for 18 h at ambient temperature. Saturated aqueous ammonium chloride solution (100 mL) was added and the mixture was stirred for 5 min. The solution was concentrated to remove excess tetrahydrofuran and diethyl ether (100 mL) was added. The aqueous phase was extracted with diethyl ether (2 x 100 mL) and the combined organic layers washed with brine (100 mL) and dried over sodium sulfate, then filtered and concentrated under reduced pressure to yield a brown slurry. Heptane (150 mL) was added, and the resultant suspension filtered. The filtrate was concentrated to give a crude brown oil. The residue was purified by normal-phase chromatography: Biotage Isolera, 330 g SiliCycle Cartridge, 0- 30% ethyl acetate in heptane over 10 CV. Product fractions were combined and concentrated to give Compound (6) (R1= C16H33) (10.3 g, 99% purity, 58% yield over 4 steps).1H NMR (400 MHz, CDCl3, ppm): d 6.00-5.88 (m, 1 H), 5.30 (dq, J = 17.2, 1.6 Hz, 1 H), 5.20 (dq, J = 10.4, 1.6 Hz, 1 H), 4.22-4.07 (m, 2 H), 3.80-3.71 (m, 1 H), 3.69- 3.64 (m, 1 H), 3.64-3.55 (m, 2 H), 3.54-3.49 (m, 1 H), 3.48-3.42 (m, 2 H), 2.17 (dd, J = 6.7, 5.6 Hz, 1 H), 1.61-1.56 (m, 2 H), 1.34-1.26 (m, 26 H), 0.91-0.87 (m, 3 H).13C NMR (100 MHz, CDCl3, ppm): d 134.8, 117.2, 77.7, 71.9, 71.1, 71.0, 63.0, 31.9, 29.8 - 29.5 (m, 9 C), 29.4, 29.3, 26.1, 22.7, 14.1

[0430] Analogous transformations were also successfully carried out using Compound (5) to yield Compound (6) wherein R1= C18H37 and R1= C18H35. Example 2: Synthetic Route 2 – Synthesis of LPE(O) Compound (10)

[0431] Synthesis of Compound (10). Each of the reactions and numbered intermediates described below refer to the corresponding reactions and intermediates in Scheme III. Route 2 synthesis relies on the Route 1 synthesis of Compound (6). Reactions (i) to (v) are therefore the same as that applied in the synthesis of Compound (6) in Example 1 and as pictured in Scheme II. 153Ref: P329WOScheme III. Exerom Compound (6).

[0432] Reaction (vi) – Formation of Compound (7): A solution of phosphorous oxychloride (725 μL, 7.78 mmol, 1.5 equiv.) in dry diethyl ether (20 mL) was stirred at 0 °C under an argon atmosphere and a solution of Compound (6) (R1= 154Ref: P329WO C16H33) (2.00 g, 5.19 mmol, 1.0 equiv.) and triethylamine (1.08 mL, 7.78 mmol, 1.5 equiv.) in dry diethyl ether (20 mL) was added dropwise over 7 minutes to give a light purple slurry [Note 1]. The reaction was stirred at 0 °C for 1 hour 40 minutes. The reaction mixture was filtered through Celite® and washed with dry tetrahydrofuran (2×20 mL) to give an off-white suspension which was filtered through Celite® again to give a clear pale-yellow solution of Compound (7) (R1= C16H33) [Note 2].31P NMR (162 MHz, CDCl3, ppm): δ 8.22 (br s, 1P).

[0433] Notes on reaction (vi): [1] Initial reaction temperature was 0°C, exotherm reached a maximum temperature of 5°C. [2] Material was processed as a solution immediately into reaction (vii) to avoid degradation.

[0434] Analogous transformations were also successfully carried out using Compound (6) to yield Compound (6) wherein R1= C18H37 and R1= C18H35.

[0435] Reaction (vii) – Formation of Compound (8): A solution of Compound (7) (R1= C16H33) (2.46 g, 5.20 mmol, 1.0 equiv.) in 1:1 dry diethyl ether:tetrahydrofuran (80 mL) was heated to 40°C under an argon atmosphere, to which was added a solution of 2-aminoethanol (543 μL, 8.99 mmol, 1.7 equiv.) and triethylamine (2.2 mL, 15.6 mmol, 3.0 equiv.) in tetrahydrofuran (12.5 mL) over 10 minutes giving a white slurry [Note 1] [Note 2]. The reaction mixture was stirred at 40°C for 30 minutes; then the heating was stopped and the reaction stirred at ambient temperature overnight [Note 3]. The reaction mixture was filtered through Celite® and washed with dry diethyl ether (40 mL) then dry tetrahydrofuran (40 mL) to give a hazy colourless solution of Compound (8) (R1= C16H33) [Note 4].1H NMR (400 MHz, CDCl3, ppm): δ 5.92 (m, 1H), 5.36 – 5.24 (m, 1H), 5.24 – 5.14 (m, 1H), 4.43 – 4.25 (m, 1H), 4.23 – 3.93 (m, 4H), 3.72 – 3.58 (m, 2H), 3.57 – 3.47 (m, 3H), 3.47 – 3.39 (m, 3H), 1.56 (br s, 2H), 1.26 (s, 26H), 0.89 (t, J = 6.8 Hz, 3H).31P NMR (162 MHz, CDCl3, ppm): δ 25.25 (br d, J = 3.9 Hz, 1P).

[0436] Notes on reaction (vii): [1] Reagent quantities were calculated by assuming 100% yield for reaction (vi). [2] Maximum reaction temperature was 44°C. 155Ref: P329WO [3] Reaction was complete by31P-NMR after the overnight hold. [4] Material was processed as a solution in reaction (viii).

[0437] Analogous transformations were also successfully carried out using Compound (7) to yield Compound (8) wherein R1= C18H37 and R1= C18H35.

[0438] Reaction (viii) – Formation of Compound (9): Aqueous hydrochloric acid (26.0 mL, 2 M, 52.0 mmol, 10 equiv.) was added in one portion to a solution of Compound (8) (R1= C16H33) (2.40 g, 5.20 mmol, 1.0 equiv.) in 1:1 diethyl ether:tetrahydrofuran (170 mL)) to give a hazy colorless solution which was stirred overnight at ambient temperature [Note 1] [Note 2]. The reaction mixture was transferred to a separating funnel and the aqueous layer removed. The organic layer was dried with sodium sulfate, filtered, and concentrated on the rotary evaporator to give a colorless gum. The material was azeotroped twice with tetrahydrofuran (50 mL then 100 mL) to give Compound (9) (R1= C16H33) (1.38 g, 82% purity by ELSD, 45% yield (3 steps)) as a white solid.1H NMR (400 MHz, CDCl3, ppm): δ 7.87 (br s, 2H), 6.02 – 5.77 (m, 1H), 5.30 (br dd, J = 17.2, 1.4 Hz, 1H), 5.23 – 5.08 (m, 1H), 4.31 (br s, 1H), 4.23 – 4.11 (m, 2H), 4.11 – 3.92 (m, 2H), 3.82 – 3.65 (m, 2H), 3.56 (m, 1H), 3.54 – 3.20 (m, 5H), 1.56 (m, 2H), 1.26 (s, 26H), 0.89 (t, J = 6.7 Hz, 3H).31P NMR (162 MHz, CDCl- 3, ppm): δ -1.23 (br s, 1P). UPLC-MS (BEH-C8, Basic, 7 minutes, 20% to 95%): ELSD: Rt = 1.927 min (81.90%), MS (ESIpos): m / z = [M+H]+480.63, MS (ESIneg): m / z = [M-H]- 478.68.

[0439] Notes on reaction (viii): [1] Reagent quantities were calculated by assuming 100% yield for reaction (viii). [2] Reaction was complete by31P-NMR after overnight hold.

[0440] Analogous transformations were also successfully carried out using Compound (8) to yield Compound (9) wherein R1= C18H37 and R1= C18H35.

[0441] Analogous transformations were also successfully carried out using trifluoroacetic acid in place of hydrochloric acid and using acetic acid in place of hydrochloric acid.

[0442] Reaction (ix) – Formation of Compound (10): 1,3- Dimethylbarbituric acid (477 mg, 3.05 mmol, 1.2 equiv.) was added to a white slurry of Compound (9) (R1= C16H33) (1.21 g, 2.52 mmol, 1.0 equiv.) in dry methanol (12 mL) 156Ref: P329WO and the resulting pale-yellow slurry was sparged with argon for 25 minutes. Palladium (0) tetrakis(triphenylphosphine) (175 mg, 0.15 mmol, 0.06 equiv.) was added in one portion and the reaction mixture heated to 51°C for 4.5 hours. A yellow suspension was observed after 5 minutes. The reaction mixture was cooled to ambient temperature and stirred over the weekend. An orange / yellow slurry was observed, and the reaction mixture was heated to 50°C; palladium (0) tetrakis(triphenylphosphine) (175 mg, 0.15 mmol, 0.06 equiv.) was charged to the flask. The reaction was heated for 4 hours then the heating was stopped and the reaction was stirred overnight [Note 1]. The reaction mixture was concentrated on the rotary evaporator to give an orange solid which was sonicated in ethyl acetate (12 mL) for 20 minutes and filtered to isolate a yellow solid which was slurried in ethyl acetate (12 mL):methanol (2.5 mL) overnight. The slurry was filtered and the filter cake was washed twice with ethyl acetate (12 mL):methanol (2.5 mL). The solid was dried on the rotary evaporator at 45°C (~30 mBar) for 40 minutes to give Compound (10) (R1= C16H33) (0.76 g, 66 wt% by Q-NMR, 97% purity by ELSD, 45% yield) as a yellow solid [Note 2].1H NMR (400 MHz, Methanol-d4:CDCl3 (1:1), ppm): δ 4.03 (br s, 2H), 3.97 – 3.78 (m, 3H), 3.55 – 3.37 (m, 4H), 3.09 (br s, 2H), 1.53 (m, 2H), 1.24 (s, 26H), 0.85 (br t, J = 6.3 Hz, 3H).31P NMR (162 MHz, Methanol- d4:CDCl3 (1:1), ppm): δ 4.77 (br s, 1P). UPLC-MS-ELSD (BEH-C8, Basic, 7 minutes, 20% to 95%): ELSD: Rt = 1.652 min (97.15%), MS (ESIpos): m / z = [M+H]+440.62, MS (ESIneg): m / z = [M-H]- 438.64.

[0443] Notes on reaction (ix): [1] After 2 hours of heating the reaction was ~95% complete by mass spectroscopy. [2] Yield is corrected for Q-NMR wt%.

[0444] Analogous transformations were also successfully carried out using Compound (9) to yield Compound (10) wherein R1= C18H37 and R1= C18H35.

[0445] Purification of Compound (10): Compound (10) (R1= C16H33) (0.720 g, 66 wt% by Q-NMR, 1.64 mmol, 1.0 equiv.) was charged to a PolyBlock vessel with chloroform (54 mL) and n-butanol (18 mL) [Note 1]. The vessel was stirred for 10 minutes at 20°C to give a hazy yellow suspension which was then heated to 60°C at 2°C / min. The contents were heated at 60°C for 15 minutes giving a clear yellow / orange solution which was then cooled to 20°C at -0.5°C / min to give a yellow suspension that 157Ref: P329WO was stirred at 20°C for 16 hours. The suspension was filtered to give a white solid on top of a layer of yellow material [Note 2]. The filter cake was washed with chloroform (10 mL) which removed some color, leaving a white upper layer and a yellow lower layer. The resultant solid was dried on the rotary evaporator at 45°C (~30 mBar) for 1.5 hours to isolate the pure Compound (10) (R1= C16H33) (347.8 mg, 99.6% purity by ELSD, 73% yield) [Note 3].1H NMR (400 MHz, Methanol-d4:CDCl3 (1:1), ppm): δ 4.03 (br s, 2H), 3.96 – 3.66 (m, 3H), 3.52 – 3.35 (m, 4H), 3.09 (br s, 2H), 1.53 (br s, 2H), 1.42 – 0.94 (s, 26H), 0.84 (br d, J = 6.2 Hz, 3H).31P NMR (162 MHz, Methanol-d4:CDCl3 (1:1), ppm): δ 4.77 (br s, 1P). UPLC-MS (BEH-C8, Basic, 7 minutes, 20% to 95%): ELSD: Rt = 1.622 min (99.64%), MS (ESIpos): m / z = [M+H]+440.59, MS (ESIneg): m / z = [M-H]- 438.64

[0446] Notes on purification: [1] The purification was carried out in a PolyBlock 4 automated parallel synthesis platform. [2] Filtration took 50 minutes. [3] Yield is corrected for Q-NMR wt% of the input material and ELSD purity of the product.

[0447] Analogous purifications were also successfully carried out of Compound (10) wherein R1= C18H37 and R1= C18H35. Example 3: Synthetic Route 3 – Synthesis of LPC(O) Compound (12)

[0448] Synthesis of Compound (12). Each of the reactions and numbered intermediates described below refer to the corresponding reactions and intermediates in Scheme IV. Route 3 synthesis relies on the Route 1 synthesis of Compound (6). Reactions (i) to (v) are therefore the same as that applied in the synthesis of Compound (6) in Example 1 and as pictured in Scheme II. 158Ref: P329WO Scheme IV. Exempl2) from Compound (6). 159Ref: P329WO

[0449] Reaction (vi-b) – Formation of Compound (7): A solution of phosphorous oxychloride (725 μL, 7.78 mmol, 1.5 equiv.) in dry diethyl ether (20 mL) was stirred at 0 °C under an argon atmosphere and a solution of Compound (6) (R1= C16H33) (2.00 g, 5.19 mmol, 1.0 equiv.) and triethylamine (1.08 mL, 7.78 mmol, 1.5 equiv.) in dry diethyl ether (20 mL) was added dropwise over 7 minutes to give a light purple slurry [Note 1]. The reaction was stirred at 0 °C for 1 hour 40 minutes. The reaction mixture was filtered through Celite® and washed with dry tetrahydrofuran (2×20 mL) to give an off-white suspension which was filtered through Celite® again to give a clear pale-yellow solution of Compound (7) (R1= C16H33) [Note 2].31P NMR (162 MHz, CDCl3, ppm): δ 8.22 (br s, 1P).

[0450] Notes on reaction (vi-b): [1] Initial reaction temperature was 0°C, exotherm reached a maximum temperature of 5°C. [2] Material was processed as a solution immediately into reaction (vii) to avoid degradation.

[0451] Analogous transformations were also successfully carried out using Compound (6) to yield Compound (7) wherein R1= C18H37 and R1= C18H35.

[0452] Reaction (vii-b) – Formation of Compound (8): A solution of Compound (7) (R1= C16H33) (2.46 g, 5.20 mmol, 1.0 equiv.) in 1:1 dry diethyl ether:tetrahydrofuran (80 mL) was heated to 40°C under an argon atmosphere, to which was added a solution of 2-aminoethanol (543 μL, 8.99 mmol, 1.7 equiv.) and triethylamine (2.2 mL, 15.6 mmol, 3.0 equiv.) in tetrahydrofuran (12.5 mL) over 10 minutes giving a white slurry [Note 1] [Note 2]. The reaction mixture was stirred at 40°C for 30 minutes; then the heating was stopped and the reaction stirred at ambient temperature overnight [Note 3]. The reaction mixture was filtered through Celite® and washed with dry diethyl ether (40 mL) then dry tetrahydrofuran (40 mL) to give a hazy colourless solution of Compound (8) (R1= C16H33) [Note 4].1H NMR (400 MHz, CDCl3, ppm): δ 5.92 (m, 1H), 5.36 – 5.24 (m, 1H), 5.24 – 5.14 (m, 1H), 4.43 – 4.25 (m, 1H), 4.23 – 3.93 (m, 4H), 3.72 – 3.58 (m, 2H), 3.57 – 3.47 (m, 3H), 3.47 – 3.39 (m, 3H), 1.56 (br s, 2H), 1.26 (s, 26H), 0.89 (t, J = 6.8 Hz, 3H).31P NMR (162 MHz, CDCl3, ppm): δ 25.25 (br d, J = 3.9 Hz, 1P). 160Ref: P329WO

[0453] Notes on reaction (vii-b): [1] Reagent quantities were calculated by assuming 100% yield for reaction (vi). [2] Maximum reaction temperature was 44°C. [3] Reaction was complete by31P-NMR after the overnight hold. [4] Material was processed as a solution in reaction (viii).

[0454] Analogous transformations were also successfully carried out using Compound (7) to yield Compound (8) wherein R1= C18H37 and R1= C18H35.

[0455] Reaction (viii-b) – Formation of Compound (9): Aqueous hydrochloric acid (26.0 mL, 2 M, 52.0 mmol, 10 equiv.) was added in one portion to a solution of Compound (8) (R1= C16H33) (2.40 g, 5.20 mmol, 1.0 equiv.) in 1:1 diethyl ether:tetrahydrofuran (170 mL)) to give a hazy colorless solution which was stirred overnight at ambient temperature [Note 1] [Note 2]. The reaction mixture was transferred to a separating funnel and the aqueous layer removed. The organic layer was dried with sodium sulfate, filtered, and concentrated on the rotary evaporator to give a colorless gum. The material was azeotroped twice with tetrahydrofuran (50 mL then 100 mL) to give Compound (9) (R1= C16H33) (1.38 g, 82% purity by ELSD, 45% yield (3 steps)) as a white solid.1H NMR (400 MHz, CDCl3, ppm): δ 7.87 (br s, 2H), 6.02 – 5.77 (m, 1H), 5.30 (br dd, J = 17.2, 1.4 Hz, 1H), 5.23 – 5.08 (m, 1H), 4.31 (br s, 1H), 4.23 – 4.11 (m, 2H), 4.11 – 3.92 (m, 2H), 3.82 – 3.65 (m, 2H), 3.56 (m, 1H), 3.54 – 3.20 (m, 5H), 1.56 (m, 2H), 1.26 (s, 26H), 0.89 (t, J = 6.7 Hz, 3H).31P NMR (162 MHz, CDCl- 3, ppm): δ -1.23 (br s, 1P). UPLC-MS (BEH-C8, Basic, 7 minutes, 20% to 95%): ELSD: Rt = 1.927 min (81.90%), MS (ESIpos): m / z = [M+H]+480.63, MS (ESIneg): m / z = [M-H]- 478.68.

[0456] Notes on reaction (viii-b): [1] Reagent quantities were calculated by assuming 100% yield for reaction (viii). [2] Reaction was complete by31P-NMR after overnight hold.

[0457] Analogous transformations were also successfully carried out using Compound (8) to yield Compound (9) wherein R1= C18H37 and R1= C18H35.

[0458] Reaction (ix-b) – Formation of Compound (11): Compound (9) (R1= C16H33) (1.13 g, 2.36 mmol, 1.0 equiv.) was stirred in iso-propanol (9.2 mL):dichloromethane (3.1 mL) to give a clear pale-yellow solution [Note 1]. Aqueous 161Ref: P329WO potassium carbonate (2.6 mL, 4 M, 10.4 mmol, 4.4 equiv.) was added in one portion, giving an off-white suspension. A solution of methyl iodide (1.3 mL, 20.9 mmol, 8.9 equiv.) in dichloromethane (6.5 mL) was charged to the flask and the reaction mixture was stirred at ambient temperature overnight [Note 2]. The reaction mixture was diluted with water (12 mL), the layers were split, and the organic layer was washed with saturated brine (12 mL), dried with sodium sulfate, and concentrated to give a brown oil / gum (1.67 g). The material was azeotroped with dichloromethane (25 mL) to give Compound (11) (R1= C16H33) (1.16 g, 94% crude isolated yield) as a brown solid.1H NMR (400 MHz, Methanol-d4:CDCl3 (1:1), ppm): δ 5.97 – 5.79 (m, 1H), 5.26 (dd, J = 17.2, 1.5 Hz, 1H), 5.13 (dd, J = 10.4, 1.3 Hz, 1H), 4.28 – 4.19 (m, 2H), 4.18 – 4.06 (m, 2H), 3.93 (dd, J = 10.9, 4.5 Hz, 1H), 3.87 (dd, J = 10.8, 5.6 Hz, 1H), 3.68 (quin, J = 5.0 Hz, 1H), 3.62 – 3.45 (m, 4H), 3.42 (dt, J = 6.8, 2.9 Hz, 2H), 3.18 (s, 9H), 1.52 (quin, J = 6.7 Hz, 2H), 1.23 (br s, 26H), 0.85 (t, J = 6.8 Hz, 3H).31P NMR (162 MHz, Methanol- d4:CDCl3 (1:1), ppm): δ -0.40 (s, 1P). UPLC-MS (BEH-C8, Long Acid, 4.5 minutes, 50% to 95%): Rt = 1.21 min, MS (ESIpos): m / z=[M+H]+522.54.

[0459] Notes on reaction (ix-b): [1] The reagent amounts were calculated based on 1.23 g of compound 9, however, the correct input mass was 1.13 g. The equivalents have been altered to reflect the actual ratios of the reagents, the intended equivalents are given in the scheme. [2] A brown solution with white solid on the walls of the flask was observed after the overnight hold, reaction was complete by mass spectroscopy.

[0460] Analogous transformations were also successfully carried out using Compound (9) to yield Compound (11) wherein R1= C18H37 and R1= C18H35.

[0461] Reaction (x-b) – Formation of Compound (12): Compound (11) (R1= C16H33) (1.16 g, 2.22 mmol, 1.0 equiv.) was added to 1,3-dimethylbarbituric acid (694 mg, 4.45 mmol, 2.0 equiv.) and methanol (17.4 mL). The reaction mixture was stirred and sparged with argon for 20 minutes; then palladium (0) tetrakis(triphenylphosphine) (257 mg, 0.22 mmol, 0.1 equiv.) was added in one portion and the reaction was heated to 55°C. 1,3-Dimethylbarbituric acid (347 mg, 2.22 mmol, 1.0 equiv.) was added in one portion and the reaction mixture heated under an argon atmosphere overnight. Heating was stopped and 1, 3-dimethylbarbituric acid (347 mg, 2.22 mmol, 1.0 equiv.) was 162Ref: P329WO added then the reaction mixture was sparged for 10 minutes with argon. Palladium (0) tetrakis(triphenylphosphine) (257 mg, 0.22 mmol, 0.1 equiv.) was added in one portion and the reaction heated to 55°C and stirred over the weekend. Heating was stopped and the reaction mixture was concentrated on the rotary evaporator to give a brown solid. The solid was sonicated in ethyl acetate (12 mL) for ~30 minutes, filtered then slurried in ethyl acetate (12 mL):methanol (2.5 mL) for 1.5 hours. The slurry was filtered and the filter cake washed with 2×(ethyl acetate (12 mL):methanol (2.5 mL)). The solid was dried at ~30 mBar and 45°C on the rotary evaporator for 2 hours to give Compound (12) (R1= C16H33) (1.04 g, 92% purity by ELSD, 89% yield) as a brown solid.1H NMR (400 MHz, Methanol-d4:CDCl3 (1:1), ppm): δ 4.33 – 4.14 (m, 2H), 3.98 – 3.76 (m, 3H), 3.64-3.53 (m, 2H), 3.50 – 3.37 (m, 4H), 3.18 (s, 9H), 1.53 (quin, J = 6.7 Hz, 2H), 1.23 (s, 26H), 0.85 (t, J = 6.5 Hz, 3H).31P NMR (162 MHz, Methanol-d4:CDCl3 (1:1), ppm): δ 0.08 (s, 1P). UPLC-MS (BEH-C8, Basic, 7 minutes, 20% to 95%): ELSD: Rt = 2.492 min (91.52%), MS (ESIpos): m / z = [M+H]+482.69, MS (ESIneg): m / z = 542.69.

[0462] Analogous transformations were also successfully carried out using Compound (11) to yield Compound (12) wherein R1= C18H37 and R1= C18H35. Example 4: Synthetic Route 4 – Alternate Synthesis of Compound (12)

[0463] Alternate synthesis of Compound (12). Each of the reactions and numbered intermediates described below refer to the corresponding reactions and intermediates in Scheme V. Route 4 synthesis relies on the Route 1 synthesis of Compound (6). Reactions (i) to (v) are therefore the same as that applied in the synthesis of Compound (6) in Example 1 and as pictured in Scheme II. 163Ref: P329WOScheme V. Ad (12) from Compound (6). 164Ref: P329WO

[0464] Preparation of Compound (31): A stirred solution of phosphorus oxychloride (14.1 mL, 152 mmol, 1.2 equiv.) in diethyl ether (90 mL) was cooled in an ice / water bath under argon [Note 1]. A solution of 2-bromoethanol (8.97 mL, 126 mmol, 1.0 equiv.) and triethylamine (21.2 mL, 152 mmol, 1.2 equiv.) in diethyl ether (90 mL) was added to the phosphorous oxychloride solution dropwise over 5 minutes [Note 2], [Note 3]. The cooling bath was removed, and the mixture was stirred at ambient temperature for 45 minutes [Note 4]. The reaction was cooled to 0°C and diluted with diethyl ether (15 mL), then filtered through celite®. The filter cake was washed with diethyl ether (2 x 15 mL) and the combined filtrates were used directly in the next step, assuming 100% yield and purity of Compound (31) [Note 5].1H NMR (400 MHz, CDCl3, ppm): 4.59 (t, J=6.4 Hz, 2 H), 3.62 (t, J=6.4 Hz, 2 H).31P NMR (162 MHz, CDCl3, ppm): ^ 7.53 (s, 1P) [Note 6].

[0465] Notes on preparation of Compound (31): [1] Internal temperature measured at 7°C. [2] Internal temperature increased to 20°C over the course of the addition. [3] A white precipitate formed over the course of the addition. The reaction mixture becomes a thick slurry if not sufficiently diluted. [4] Reaction monitored by1H and31P NMR. [5] Filtrate appears as a hazy, light-yellow solution. [6] Several minor peaks seen in31P NMR which correspond to unreacted phosphorus oxychloride, potential adducts with triethylamine and other phosphorus species.

[0466] Reaction (vi-a) – Formation of Compound (9a): A solution of Compound (6) (R1= C18H37) (6.95 g, 18.1 mmol, 1.0 equiv.) and triethylamine (2.52 mL, 18.1 mmol, 1.0 equiv.) in diethyl ether (140 mL) was added via addition funnel over 25 minutes to a stirred solution of Compound (31) (30.6 g, 126 mmol, 7.0 equiv.) in diethyl ether (695 mL) at 0°C under argon was added [Note 1], [Note 2]. The reaction was stirred at 25°C overnight. A solution of triethylamine (15.1 mL, 108 mmol, 6.0 equiv.) in diethyl ether (50 mL) was added dropwise over 5 minutes to give an off-white slurry [Note 3]. The reaction was then stirred at ambient temperature overnight. The reaction mixture was diluted with diethyl ether (20 mL) and filtered through celite, washing with diethyl ether (2 x 10 mL). The filtrate was passed through a fresh pad of celite to give a 165Ref: P329WO clear, pale-yellow solution. The solution was washed with aqueous hydrochloric acid (2 M, 2 x 20 mL) then brine (20 mL), dried over sodium sulfate, and filtered. Water (2.10 mL, 117 mmol, 6.5 equiv.) and formic acid (3.3 mL, 77.5 mmol, 4.8 equiv.) was added to the crude solution and the mixture was stirred at ambient temperature overnight. Additional formic acid (2.04 mL, 54.2 mmol, 3.0 equiv.) and water (3.25 mL, 181 mmol, 10.0 equiv.) were added and the mixture was stirred for 1 hour [Note 4]. The organic phase was washed with water (2 x 200 mL) and the aqueous layers were back- extracted with diethyl ether (200 mL). The aqueous layers were dried over sodium sulfate, filtered, and concentrated. The crude residue was purified by normal phase chromatography (Biotage Isolera, 330 g Siliasep cartridge; eluent: 0-12% methanol in dichloromethane over 10 CV). The product fractions were combined to give Compound (9a) (R1= C18H37) as two solids: solid 1 (1.67 g, 92% purity by ELSD) and solid 2 (1.27 g, 50% purity by ELSD). Both batches were light brown gums and the combined yield corrected for ELSD purity was ~21% [Note 5].1H NMR (400 MHz, CDCl3, ppm): δ 5.92 (m, 1H), 5.39 – 5.25 (m, 1H), 5.20 (br d, J = 10.3 Hz, 1H), 4.30 (m, 2H), 4.24 – 3.95 (m, 4H), 3.73 (m, 1H), 3.62 – 3.49 (m, 4H), 3.48 – 3.31 (m, 2H), 1.55 (m, 2H), 1.26 (s, 30H), 0.89 (t, J = 6.8 Hz, 3H). [Note 6].31P NMR (162 MHz, CDCl3, ppm): ^ 0.14 (s, 1P). UPLC-MS (BEH-C18, Basic, 7 minutes, 2% to 95%): ELSD: Rt = 3.190 min (92.15%), MS (ESIpos): m / z = [M+H]+1:1571.75:573.73 (79Br:81Br).

[0467] Notes on reaction (vi-a): [1] The experiment reported here used a different batch than the one reported in the preparation of Compound (31) above, hence the difference in scale. [2] Thin brown suspension formed over course of addition. No exotherm noted. [3] Additional triethylamine was added to push reaction to completion. [4] Additional formic acid / water were added to push hydrolysis to completion. [5] Data reported for solid 1. [6] Exchangeable phosphoric acid proton not observed as D2O was added to this sample.

[0468] Reaction (vii-a) – Formation of Compound (11): Silver(I) oxide (678 mg, 2.93 mmol, 1.0 equiv.) followed by a solution of trimethylamine (2.0 M in tetrahydrofuran, 14.6 mL, 29.3 mmol, 10.0 equiv.) were added to a stirred solution of 166Ref: P329WO Compound (9a) (R1= C18H37) (1.67 g, 2.93 mmol, 1.0 equiv.) in tetrahydrofuran (17 mL) and the mixture was stirred at ambient temperature overnight [Note 1], [Note 2]. The reaction mixture was concentrated to give a crude residue, which was suspended in methanol and filtered through a pad of Celite®. The filter cake was washed with methanol and the combined filtrates were concentrated to dryness to give Compound (11) (R1= C18H37) (1.06 g, 64% yield) as a thick yellow gel [Note 3].1H NMR (400 MHz, Methanol-d4, ppm): ^ 6.00 ‐ 5.88 (m, 1 H), 5.30 (m, 1 H), 5.14 (m, 1 H), 4.32 - 4.23 (m, 2 H), 4.17 (d, J=5.1 Hz, 2 H), 4.00 – 3.86 (m, 2 H), 3.71 (quin, J=5.2 Hz, 1 H), 3.65 - 3.61 (m, 2 H), 3.60 - 3.49 (m, 2 H), 3.49 - 3.43 (m, 2 H), 3.22 (s, 9 H), 1.56 (quin, J= 6.8 Hz, 2 H), 1.40 - 1.28 (m, 30 H), 0.90 (t, J=6.7 Hz, 3 H).13C NMR (100 MHz, Methanol- d4, ppm): ^ 136.8, 117.2, 78.9, 78.8, 72.8, 72.4, 71.7, 66.7, 66.6, 60.5, 60.5, 54.9, 54.8, 54.8, 33.2, 30.9 (s, 9 C), 30.8, 30.6, 27.4, 23.9, 14.6.31P NMR (162 MHz, CDCl3, ppm): d -0.34 (s, 1 P). UPLC-MS (BEH-C8, Basic, 7 minutes, 20% to 95%): ELSD: Rt = 3.222 min (93.30%), MS (ESIpos): m / z = [M+H]+550.82.

[0469] Notes on reaction (vii-a): [1] The flask is covered in foil during the reaction to exclude light as a precaution given the sensitivity of silver (I) salts to visible light. [2] The reaction mixture is initially a colorless solution, but it quickly goes to a dark brown suspension once the amine is added. [3] Yield is corrected for the ELSD purity of the starting material and product.

[0470] Reaction (viii-a) – Formation of Compound (12): 1,3- Dimethylbarbituric acid (364 mg, 2.33 mmol, 1.2 eq.) was added to a stirred solution of Compound (11) (R1= C18H37) (1.06 g, 1.93 mmol, 1.0 eq.) in methanol (11 mL) and the mixture degassed with argon for 20 minutes. Tetrakis(triphenylphosphine)palladium(0) (44.6 mg, 38.6 µmol, 0.02 eq.) was added to the Compound (11) mixture, and the mixture was stirred at 50°C for 3 hours [Note 1], [Note 2]. The reaction mixture was concentrated on the rotary evaporator to give an orange solid. The residue was suspended in ethyl acetate (10 mL) and sonicated for 30 mins to give a slurry, which was filtered to give a bright orange solid. The isolated solid was stirred in 5:1 ethyl acetate: methanol (12 mL) to give a slurry which was filtered. The filter cake was washed twice with 5:1 ethyl acetate: methanol (12 mL) to give a white solid with flecks 167Ref: P329WO of orange. The isolated solid was dried on the rotary evaporator at 45°C for 35 minutes [Note 3]. The dried solid was crushed to give Compound (12) (R1= C18H37) as a light orange powder (571.8 mg, 56% yield, 97% purity by ELSD).1H NMR (400 MHz, Methanol-d4, ppm): ^ 4.36 – 4.21 (m, 2H), 3.98 – 3.80 (m, 3H), 3.67 – 3.60 (m, 2H), 3.55 – 3.40 (m, 4H), 3.22 (s, 9H), 1.57 (quin, J = 6.7 Hz, 2H), 1.41 – 1.19 (s, 30H), 0.95 – 0.83 (t, 3H) [Note 4].13C NMR (100 MHz, Methanol-d4, ppm): ^ 73.1, 72.9, 71.2, 71.1, 68.7, 68.6, 60.6, 60.5, 54.9, 54.8, 54.8, 33.2, 30.9 (s, 9 C), 30.8, 30.6, 27.4, 23.9, 14.6.31P NMR (162 MHz, Methanol-d4, ppm): ^ -0.05 (s, 1P). UPLC-MS (BEH-C8, Basic, 7 minutes, 20% to 95%), ELSD: Rt = 2.846 min (97.07%), MS (ESIpos): m / z= [M+H]+510.82.

[0471] Notes on reaction (viii-a): [1] The reaction was monitored by UPLC-MS (BEH-C8, Acidic, 5 minutes, 50% to 95%). Material not UV active so mass spectroscopy used for reaction monitoring. [2] The reaction mixture goes from an orange suspension to a clear orange solution over the course of the reaction. [3] An initial trituration had been carried out in an ethyl acetate / dichloromethane / methanol solvent system, however, recovery was low (200 mg), so the liquors were concentrated and a second trituration was carried out (described in the procedure). The solids isolated from both triturations were combined for drying on the rotary evaporator. [4] Hydroxyl proton not observed. Example 5: Synthetic Route 5 – Alternate Synthesis of Compound (10)

[0472] Alternate synthesis of Compound (10). Each of the reactions and numbered intermediates described below refer to the corresponding reactions and intermediates in Scheme VI. Route 5 synthesis relies on the Route 1 synthesis of Compound (6). Reactions (i) to (v) are therefore the same as that applied in the synthesis of Compound (6) in Example 1 and as pictured in Scheme II. 168Ref: P329WOScheme VI. AlternateCompound (6). 169Ref: P329WO

[0473] Preparation of Compound (16): Tetrazole (0.45 M solution in acetonitrile, 49.0 mL, 22.1 mmol, 1.25 equiv.) was added dropwise over 5 mins to a stirred solution of bis(2-cyanoethyl)-N,N-diisopropylphosphoramidite (Compound (30)) (6.65 g, 22.1 mmol, 1.25 equiv.) in tetrahydrofuran (50 mL) at room temperature under argon. The mixture stirred was at room temperature [Note 1]. A solution of (9H-fluoren- 9-yl)methyl N-(2-hydroxyethyl)carbamate (5.00 g, 17.7 mmol, 1.0 equiv.) was then added dropwise over 5 mins and the resulting mixture stirred at room temperature for 1 h [Note 2]. The reaction mixture was diluted with dichloromethane (100 mL) and transferred to a separating funnel. The organic layers were washed with saturated aqueous sodium bicarbonate solution (50 mL) and the aqueous layer was back- extracted with dichloromethane (100 mL) [Note 3]. The combined organic layers were dried over sodium sulfate, filtered, and concentrated to give 10.8 g of an orange oil. The residue was purified by normal phase chromatography: Biotage Isolera, 80 g SiliCycle Cartridge, 10-50% ethyl acetate +1% triethylamine in heptane + 1% triethylamine over 12 CV [Note 4]. Product fractions were combined and concentrated to give Compound (16) (5.83 g, 97% purity, 66% yield) as an extremely viscous colourless oil [Note 5]; [Note 6].1H NMR (400 MHz, CDCl3, ppm): 7.81-7.77 (m, 2 H), 7.62 (br d, J=7.5 Hz, 2 H), 7.45-7.40 (m, 2 H), 7.37-7.32 (m, 2 H), 5.26 (br s, 1 H), 4.43 (d, J=7.0 Hz, 2 H), 4.29 - 4.20 (m, 1 H), 3.94-3.57 (m, 6 H), 3.45 (q, J=5.3 Hz, 2 H), 2.63 (t, J=6.3 Hz, 2 H), 1.21 (t, J=6.6 Hz, 12 H).13C NMR (100 MHz, CDCl3, ppm): 156.4 (s, 1 C), 144.0 (s, 2 C), 141.3 (s, 2 C), 127.7 (s, 2 C), 127.0 (s, 2 C), 125.0 (s, 2 C), 120.0 (s, 2 C), 117.6, 66.7, 62.7 (d, J = 16.9 Hz, 1 C), 58.4 (d, J = 19.8 Hz, 1 C), 47.2, 43.1 (d, J=12.5 Hz, 2 C), 42.1 (d, J = 7.3 Hz, 1 C), 25.55 – 24.07 (m, 4 C), 20.4 (d, J = 7.3 Hz, 1 C). UPLC-MS (BEH-C18, Basic, 2 minutes, 50% to 95%): Rt = 0.83 min (96.64%), MS (ESIpos): m / z = [M+Na]+ 506.3.

[0474] Reaction (vi-c) – Formation of Compound (13): Tetrazole (0.45 M in acetonitrile, 21 mL, 1.1 equiv.) was added to a stirred solution of Compound (16) (4.52 g, 8.61 mmol, 1.0 equiv.) and Compound (6) (R1= C16H33) (3.10 g, 8.61 mmol, 1.0 equiv.) in dichloromethane (45 mL) at room temperature. The resulting solution was stirred at room temperature for 3 hours. The solution was quenched by addition of saturated aqueous sodium bicarbonate solution (50 mL) and the aqueous phase was 170Ref: P329WO back-extracted with dichloromethane (2 x 20 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated to give Compound (13) (R1= C16H33) (7.21 g, 71.5% purity, Crude) as a yellow oil [Note 7].1H NMR (400 MHz, CDCl3, ppm): 7.81-7.74 (m, 2 H), 7.61 (br d, J = 7 Hz, 2 H), 7.45-7.38 (m, 2 H), 7.35- 7.30 (m, 2 H), 5.99-5.85 (m, 1 H), 5.58-5.40 (m, 1 H), 5.31-5.25 (m, 1 H), 5.23-5.15 (m, 1 H), 4.41-4.38 (m, 2 H), 4.35 - 4.09 (m, 6 H), 4.07-3.83 (m, 2 H), 3.79-3.35 (m, 8 H), 2.80-2.72 (m, 1 H), 2.66-2.59 (m, 1 H), 1.61-1.50 (m, 2 H), 1.26 (s, 26 H), 0.89 (t, J = 7 Hz, 3 H). UPLC-MS (BEH-C8, Basic, 4.5 minutes, 50% to 95%): Rt = 3.56 min (71.52%), MS (ESIpos): m / z = [M+Na]+761.56.

[0475] Notes on reaction (vi-c): [1] During the addition of the tetrazole solution, a precipitate formed. [2] Depending on the commercial source of (9H-fluoren-9-yl)methyl N-(2- hydroxyethyl)carbamate the mixture is either colorless or light-yellow. There is no observable effect on yield or purity of the isolated material. [3] The P(III) intermediate is extremely acid sensitive; any aqueous washings must be done with a basic solution. [4] Use of triethylamine in both column solvents to fully neutralize the silica solid phase is ideal. [5] Once isolated the material has been stored under argon in a freezer for several days. [6] Due to concerns about the scalability of the reaction, the reaction was kept at 5 g scale with respect to the (9H-fluoren-9-yl)methyl N-(2-hydroxyethyl)carbamate and carried out 3 times to generate the required amounts of compound 8. [7] A small amount of the oxidized Compound (14) is also observed in the UPLC at this stage.

[0476] Analogous transformations were also successfully carried out using Compound (6), Compound (30), and Compound (16) to yield Compound (13) wherein R1= C18H37 and R1= C18H35.

[0477] Reaction (vii-c) – Formation of Compound (14): Meta- chloroperbenzoic acid (3.29 g, 13.4 mmol, 70% purity, 1.55 equiv.) was added portionwise over 10 minutes to a stirred solution of Compound (13) (R1= C16H33) (6.36 171Ref: P329WO g, 8.61 mmol, 1.0 equiv) in tetrahydrofuran (80 mL) at 0 °C. The reaction was stirred at 0 °C for 10 minutes and then warmed to room temperature and stirred for 1 hr. The solution was partitioned between dichloromethane (80 mL) and saturated aqueous sodium bicarbonate solution (80 mL) and the layers separated. The organic layer was washed with saturated aqueous sodium bicarbonate solution (2 x 30 mL). The combined aqueous layers were back-extracted with dichloromethane (2 x 30 mL) and the combined organic layers were washed with brine (60 mL), then dried over sodium sulfate, filtered, and concentrated. The crude residue was purified by normal-phase chromatography: Biotage Isolera, 80 g SiliCycle Cartridge, 10-100% ethyl acetate in heptane over 16 CV. Product fractions were combined and concentrated to give Compound (14) (R1= C16H33) (3.00 g, 96.7% purity, 45% yield, 2 steps) as a yellow oil.1H NMR (400 MHz, CDCl3, ppm): 7.78 (d, J = 7.6 Hz, 2H), 7.62 (br d, J = 7.3 Hz, 2H), 7.41 (t, J = 7.3 Hz, 2H), 7.33 (td, J = 7.6, 1.1 Hz, 2H), 5.98 – 5.81 (m, 1H), 5.61 – 5.42 (m, 1H), 5.35 – 5.23 (m, 1H), 5.23 – 5.14 (m, 1H), 4.42 (br d, J = 6.8 Hz, 2H), 4.34 – 4.07 (m, 9H), 3.78 – 3.62 (m, 1H), 3.61 – 3.27 (m, 6H), 2.74 (t, J = 6.2 Hz, 2H), 1.52 (m, 2H), 1.37 – 1.16 (m, 26H), 0.94 – 0.81 (m, 3H).13C NMR (100 MHz, CDCl3, ppm): 156.4, 143.9 (s, 2 C), 141.3 (s, 2 C), 134.5 (d, J = 1 Hz, 1 C), 127.7 (s, 2 C), 127.1 (s, 2 C), 125.1 (s, 2 C), 120.0 (s, 2 C), 117.5, 117.5, 77.2, 76.2 (d, J = 5.9 Hz, 1 C), 71.9, 71.3, 69.2 (d, J = 4.4 Hz, 1 C), 67.8 (d, J = 5.9 Hz, 1 C), 67.4, 66.9 (d, J = 2.2 Hz, 1C), 61.9 (d, J = 5.9 Hz, 1 C), 47.2, 41.3 (d, J = 5.9 Hz, 1 C), 31.9, 29.7-29.6 (m, 8 C), 29.5, 29.4, 26.1, 22.7, 19.6 (d, J = 5.9 Hz, 1 C), 14.1.31P NMR (162 MHz, CDCl3, ppm): -1.36 (s, 1 P).

[0478] Analogous transformations were also successfully carried out using Compound (13) to yield Compound (14) wherein R1= C18H37 and R1= C18H35.

[0479] Reaction (viii-c) – Formation of Compound (15): A stirred solution of Compound (14) (R1= C16H33) (10.26 g, 13.3 mmol, 1.0 equiv.) in dichloromethane (90 mL) was degassed with argon for 10 mins. Palladium tetrakis(triphenylphosphine) (3.38 g, 2.93 mmol, 0.22 equiv.) and 1,3-dimethylbarbituric acid (13.83 g , 88.7 mmol, 6.66 equiv.) were added and the mixture heated to 40 °C for 5 hours. The solution was cooled to room temperature, then filtered through Celite®, washing with dichloromethane. The filtrate was concentrated to give an orange solid. The crude 172Ref: P329WO residue was purified by normal-phase chromatograpy: Biotage Isolera, 330 g SiliCycle Cartridge, 0-5% methanol in dichloromethane over 16 CV. Product fractions were combined and concentrated to give Compound (15) (R1= C16H33) (8.66 g, 94.5% purity, 91% yield) as a dark-red solid.1H NMR (400 MHz, CDCl3, ppm): 7.80-7.75 (m, 2 H), 7.61 (br d, J = 7.5 Hz, 2 H), 7.44-7.38 (m, 2 H), 7.37-7.30 (m, 2 H), 5.60-5.49 (m, 1 H), 4.47-4.38 (m, 2 H), 4.31-4.08 (m, 7 H), 4.04-3.96 (m, 1 H), 3.56-3.48 (m, 2 H), 3.48-3.37 (m, 4 H), 2.75 (t, J = 6.2 Hz, 2 H), 2.18 (s, 1 H), 1.59-1.51 (m, 2 H), 1.26 (s, 26 H), 0.93- 0.86 (m, 3 H).13C NMR (100 MHz, CDCl3, ppm): 156.5, 143.9 (s, 2 C), 141.3 (s, 2 C), 127.7 (s, 2 C), 127.1 (s, 2 C), 125.1 (s, 2 C), 120.0 (s, 2 C), 116.5, 71.9, 70.6, 69.8, 69.4 (d, J = 5.9 Hz, 1 C), 67.5 (d, J = 6.6 Hz, 1 C), 66.9, 62.1 (d, J = 5.1 Hz, 1 C), 47.2, 41.3 (d, J = 5.1 Hz, 1 C), 31.9, 30.9, 29.7-29.6 (m, 5 C), 29.6, 29.6, 29.5, 29.5, 29.3, 26.0, 22.7, 19.7 (d, J = 5.9 Hz, 1 C), 14.1.31P NMR (162 MHz, CDCl3, ppm): -0.96 (s, 1 P). UPLC-MS (BEH-C8, Basic, 4.5 minutes, 50% to 95%): Rt = 2.72 min (94.50%), MS (ESIpos): m / z = [M+H]+715.61.

[0480] Analogous transformations were also successfully carried out using Compound (14) to yield Compound (15) wherein R1= C18H37 and R1= C18H35.

[0481] Reaction (ix-c) – Formation of Compound (10): Piperidine (691 μL, 7.00 mmol, 10 equiv.) was added to a stirred solution of Compound (15) (R1= C16H33) (500 mg, 700 μmol, 1.0 equiv.) in N.N-dimethylformamide (2.5 mL). The mixture was stirred at room temperature for 10 minutes. The reaction mixture was loaded directly onto a reverse-phase column and purified [Note 1]. The material was purified by reverse-phase chromatography: Biotage Isolera, 40 g SiliCycle C18Cartridge, 10- 100% (acetonitrile + 0.1% formic acid) in water (+0.1% Formic acid) over 15 CV). The gradient was held at 80% acetonitrile whilst product was eluting [Note 2]. Product fractions were combined and concentrated to give Compound (10) (R1= C16H33) (70 mg, 97.3% purity, 22% yield) as a colourless solid [Note 3]. Multiple batches were combined to give 557 mg of Compound (10) (R1= C16H33).1H NMR (400 MHz, CDCl3, ppm): 8.28 (br s, 2 H) 4.50-4.11 (m, 4 H) 4.08–3.84 (m, 2 H) 3.82-3.60 (m, 1 H) 3.56- 3.38 (m, 4 H) 3.24 (br s, 2 H) 1.60-1.48 (m, 2 H) 1.26 (s, 26 H) 0.92-0.86 (m, 3 H).31P NMR (162 MHz, CDCl3, ppm): 0.25 (s, 1 P), -0.45 (s, 1 P). UPLC-MS (CSH-C18, Acidic, 4.5 minutes, 2% to 95%): ELSD: Rt = 3.341 min (97.26%), MS (ESIpos): m / z = [M+H]+ 173Ref: P329WO 440.66. UPLC-MS (CSH-C18, Basic, 4.5 minutes, 40% to 80%): ELSD: Rt = 2.063 min (24.04%), MS (ESIpos): m / z = [M+H]+ 440.62; Rt = 2.103 min (73.24%), MS (ESIpos): m / z = [M+H]+ 440.66.

[0482] Notes on reaction (ix-c): [1] Minimization of reaction time is helpful to prevent excess product degradation. The reverse-phase column should be pre-equilibrated to reduce setup time. [2] The product is not UV active and cannot be seen by ELS whilst on the column. The product appears to elute as a very broad peak despite running well by UPLC. [3] Due to difficulties scaling the reaction, the required amounts of target material were synthesised in 500 mg batches.

[0483] Analogous transformations were also successfully carried out using Compound (15) to yield Compound (10) wherein R1= C18H37 and R1= C18H35. Example 6: Synthetic Route 6 – Alternate Synthesis of Compound (12)

[0484] Alternate synthesis of Compound (12). Each of the reactions and numbered intermediates described below refer to the corresponding reactions an intermediates in Scheme VII. Route 6 does not rely on Compound (6) but rather utilizes different raw materials. 174Ref: P329WO Scheund (12). 175Ref: P329WO

[0485] Preparation of Compound (17) (R1= C16H31D2): Lithium aluminum deuteride (774 mg, 18.48 mmol) was added to a solution of methyl palmitate (5.0 g, 18.48 mmol) in diethyl ether (10 mL, 10V) at 0-5°C and the reaction mixture was stirred for 30 min at the same temperature. After removal from the ice-bath, the reaction mixture was allowed to stir at room temperature over a period of 30 min. Reaction completion was monitored by TLC (10% ethyl acetate in n-hexane). Once complete, the reaction mixture was quenched with saturated ammonium chloride solution (100 mL) at 0-5°C and stirred for 5 min at the same temperature. The resulting mixture was filtered through a celite bed and the solids were washed with ethyl acetate (20 mL). The filtrate was extracted with ethyl acetate (3X100 mL) and the separated organic layer was dried over sodium sulphate and concentrated. The crude product obtained upon evaporation of volatiles was purified through a silica gel (60-120) column (8% ethyl acetate in n-hexane) to give Compound (17) (R1= C16H31D2) as a white solid (3.3 g, 74%).1H NMR (300 MHz, DMSO-d6) δ (ppm): 4.24 (s, 1H), 1.40-1.35 (m, 2H), 1.22- 1.21 (m, 26H), 0.85-0.81 (m, 3H). GC-MS m / z (M-H2O): 226 (Calculated molecular weight – 244.45). HPLC purity: 98.76%.

[0486] An analogous transformation was also successfully carried out using methyl stearate in diethyl ether and lithium aluminum deuteride to yield Compound (17) wherein R1= C18H35D2. Likewise, an analogous transformation was also successfully carried out using methyl oleate in diethyl ether and lithium aluminum deuteride to yield Compound (17) wherein R1= C18H33D2

[0487] Reaction (1-a) – Formation of Compound (19): Compound (17) (R1= C16H31D2) (1.04 g, 4.3 mmol) was added to a solution of (2R)-(-)-Glycidyl tosylate (Compound (18)) (1.0 g, 4.3 mmol) in dichloromethane (20 mL, 20V) at room temperature (25-30 °C). Boron trifluoride etherate (0.26 mL, 0.86 mmol) was added to the above reaction mixture at 0-5°C and stirring was continued for 10 min at the same temperature. After removal from the ice-bath, the reaction mixture was allowed to stir at room temperature over a period of 48h. Reaction completion was monitored by TLC (30% ethyl acetate in n-hexane). After completion, the reaction mixture was concentrated. The crude product obtained upon evaporation of volatiles was purified through a silica gel (230-400) column (15% ethyl acetate in n-hexane) to give 176Ref: P329WO Compound (19) (R1= C16H31D2) as a white solid (1.05 g, 51 %).1H NMR (300 MHz, DMSO-d6) δ (ppm): 7.77 (d, J=8.1 Hz, 2H), 7.47 (d, J= 7.8 Hz, 2H), 5.21 (d, J= 5.1 Hz, 1H), 3.97 (dd, J= 9.9 Hz, 3.6 Hz, 1H), 3.85 (dd, J= 9.6 Hz, 6 Hz, 1H), 3.75-3.70 (m, 1H), 3.29-3.17 (m, 2H), 2.41 (s, 3H), 1.38-1.35 (m, 2H), 1.30-1.23 (m, 26H), 0.84 (t, J=6 Hz, 3H). MS m / z (M+Na): 495.5 (Calculated molecular weight – 472.70).

[0488] Analogous transformations were also successfully carried out using Compound (17) and Compound (18) to yield Compound (19) wherein R1= C18H35D2 and R1= C18H33D2.

[0489] Reaction (1-b) – Formation of Compound (20): Benzyl 2,2,2- trichloroacetimidate (0.8 mL, 0.07 mmol) and trifluoromethanesulfonic acid were added to a solution of Compound (19) (R1= C16H31D2) (1.0 g, 4.23 mmol) in diethyl ether (20mL, 20V) at 0-5°C. The reaction mixture was allowed to stir at room temperature over a period of 16 h. Reaction completion was monitored by TLC (30% ethyl acetate in n-hexane). After completion, the reaction mixture was concentrated. The crude product obtained upon evaporation of volatiles was purified through a silica gel (60-120) column (10% ethyl acetate in n-hexane) to give Compound (20) (R1= C16H31D2) as a pale- yellow liquid (0.95 g, 80%).1H NMR (300 MHz, DMSO-d6) δ (ppm): 7.77 (d, J= 8.4 Hz, 2H), 7.45(d, J= 8.1 Hz, 2H), 7.34-7.22 (m, 5H), 4.50-4.48 (m, 2H), 4.17-4.14 (m, 1H), 4.04-3.99 (m, 1H), 3.70-3.69 (m, 1H), 3.38-3.36 (m, 2H), 2.41 (s, 3H), 1.41-1.36 (m, 2H) 1.31-1.22 (m, 26H), 0.88-0.82 (m, 3H). MS m / z (M+ Na): 585.5 (Calculated molecular weight-562.84).

[0490] Analogous transformations were also successfully carried out using Compound (19) to yield Compound (20) wherein R1= C18H35D2 and R1= C18H33D2. When R1= C18H33D2, 4-methoxybenzyl 2,2,2-trichloroacetimidate and lanthanum triflate were used in place of benzyl 2,2,2-trichloroacetimidate and trifluoromethanesulfonic acid.

[0491] Reaction (1-c) – Formation of Compound (21): Cesium acetate (1.27 g, 6.66 mmol) was added to a solution of Compound (20) (R1= C16H31D2) (1.0g, 1.77 mmol) in DMSO (20 mL, 20V) and DMF (5 mL, 5V) at room temperature (25-30°C) and the reaction mixture heated to 60°C over a period of 16 hours. Reaction completion was monitored by TLC (20% ethyl acetate in n-hexane). After completion and allowing 177Ref: P329WO the reaction mixture to reach room temperature, it was diluted with water (30 mL) and extracted with MTBE (3X50 mL). The separated organic layer was dried over sodium sulphate and concentrated. The crude product obtained upon evaporation of volatiles was purified through silica gel (60-120) column (8% ethyl acetate in n-hexane) to yield Compound (21) (R1= C16H31D2) as a colorless liquid (680 mg, 80%).1H NMR (400 MHz, DMSO-d6) δ (ppm): 7.34-7.11 (m, 5H), 4.63 (s, 2H), 4.23 (dd, J =11.6 Hz, J =4 Hz, 1H), 4.09 (dd, J =12 Hz, J =6 Hz, 1H), 3.76 (quint, J =5.2 Hz, 1H), 3.51-3.46 (m, 2H), 2.00 (s, 3H), 1.51-1.50 (m, 2H), 1.39-1.26 (m, 26H), 0.87 (t, J =7.2 Hz, 3H). MS m / z (M+ Na): 473.6 (Calculated molecular weight-450.69).

[0492] Analogous transformations were also successfully carried out using Compound (20) to yield Compound (21) wherein R1= C18H35D2 and R1= C18H33D2.

[0493] Reaction (1-d) – Formation of Compound (22): Sodium methoxide (0.12 g, 2.21 mmol) was added to a solution of Compound (21) (R1= C16H31D2) (1.0 g, 2.21 mmol) in methanol (10 mL, 10V) at room temperature (25-30°C). The reaction mixture was heated to 45°C over a period of 16 hours. Reaction completion was monitored by TLC (30% ethyl acetate in n-hexane). After completion and allowing the reaction mixture to reach room temperature, it was neutralized with 1.5N HCl solution, diluted with water (10 mL), and extracted with ethyl acetate (3X25 mL). The separated organic layer was then dried over sodium sulphate and concentrated. The crude product obtained upon evaporation of volatiles were purified through silica gel (60-120) column (20% ethyl acetate in n-hexane) to give Compound (22) (R1= C16H31D2) as a colorless liquid (790 mg, 85 %).1H NMR (400 MHz, CDCl3) δ (ppm): 7.35-7.28 (m, 5H), 4.73-4.61 (m, 2H), 3.77-3.73 (m, 1H), 3.69-3.64 (m, 2H), 3.59 (dd, J = 9.6 Hz, 4.8 Hz, 1H), 3.53 (dd, J = 9.6 Hz, 5.2 Hz, 1H), 1.59-1.53 (m, 2H), 1.38-1.25 (m, 26H), 0.87 (t, J=6.8 Hz, 3H). MS m / z (M+ Na): 431.6 (Calculated molecular weight-408.65).

[0494] Analogous transformations were also successfully carried out using Compound (21) to yield Compound (22) wherein R1= C18H35D2 and R1= C18H33D2.

[0495] Reaction (1-e) – Formation of Compound (23): Triethylamine (0.6 mL, 4.3 mmol) and ethylene chlorophosphate (0.3mL, 3.22 mmol) were added to a solution of Compound (22) (R1= C16H31D2) (1.0 g, 2.15 mmol) in toluene (10 mL, 10V) at 0-5°C and the reaction mixture was stirred for 10 min at the same temperature. The 178Ref: P329WO reaction mixture was then allowed to stir at room temperature over a period of 2 hours. Reaction completion was monitored by TLC (40% ethyl acetate in n-hexane). After completion, the resulting reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (2X50 mL). The separated organic layer was dried over sodium sulphate and concentrated to yield Compound (23) as a colorless liquid (1.2 g crude). Compound (23) (R1= C16H31D2) was immediately used for reaction (1-f) without further purification.1H NMR (400 MHz, CDCl3) δ (ppm): 7.38-7.27 (m, 5H), 4.73-4.64 (m, 2H), 4.44-4.39 (m, 2H), 4.36-4.24 (m, 2H), 4.22-4.20 (m, 2H), 3.78-3.76 (m, 1H), 3.53-3.51 (m, 2H), 1.54-1.51 (m, 2H), 1.32-1.25 (m, 26H), 0.87 (t, J=6.8 Hz, 3H). MS m / z (M+H): 515.6 (Calculated molecular weight-514.67).

[0496] Analogous transformations were also successfully carried out using Compound (22) to yield Compound (23) wherein R1= C18H35D2 and R1= C18H33D2.

[0497] Reaction (1-f) – Formation of Compound (24): Trimethylamine in THF (44.1 mL, 97.0 mmol) was added to a solution of Compound (23) (R1= C16H31D2) (1.0 g, 1.94 mmol) in ACN (20 mL, 20V) at 25°C. The reaction mixture was allowed to stir at 70°C over a period of 16 hours. Reaction completion was monitored by TLC (80% methanol / 20% ethyl acetate / 1mL aqueous ammonia). After completion and allowing the reaction mixture to reach room temperature, it was concentrated under vacuum to yield the crude product Compound (24) (R1= C16H31D2) as a pale-yellow liquid (1.1 g crude). Crude Compound (24) was used for reaction (1-g) without further purification.1H NMR (400 MHz, CDCl3) δ (ppm): 7.34-7.22 (m, 5H), 4.70-4.61 (m, 2H), 4.19-4.17 (m, 1H), 3.96-3.89 (m, 2H), 3.76-3.67 (m, 2H), 3.65-3.53 (m, 4H), 3.10 (s, 9H), 1.58-1.52 (m, 2H), 1.35-1.24 (m, 26H), 0.88-0.87 (m, 3H). MS m / z (M+H): 574.7 (Calculated molecular weight-573.78).

[0498] Analogous transformations were also successfully carried out using Compound (23) to yield Compound (24) wherein R1= C18H35D2 and R1= C18H33D2.

[0499] Reaction (1-g) – Formation of Compound (12):10% Pd / C (200 mg) was added to a solution of Compound (24) (R1= C16H31D2) (1.0 g, 1.74 mmol) in methanol (20V) at 25°C. The reaction mixture was stirred under hydrogen atmosphere (392.266 kPa) over a period of 6 hours. Reaction completion was monitored by TLC (80% methanol / 20% ethyl acetate / 1mL aqueous ammonia). After completion, the 179Ref: P329WO resulting reaction mixture was filtered through celite and the solids were washed with excess methanol. The filtrate and wash were then concentrated. The crude product obtained upon evaporation of volatiles was purified through MB-20 resin column to give Compound (12) (R1= C16H31D2) as a thick liquid. The thick liquid was crystallized from diethyl ether and acetonitrile to yield pure Compound (12) (R1= C16H31D2) as an off white solid (135 mg, 16%).1H NMR (400 MHz, CDCl3-CD3OD-4:1) δ (ppm): 4.22-4.18 (m, 2H), 3.92-3.76 (m, 3H), 3.52-3.50 (m, 2H), 3.39-3.38 (m, 2H), 3.13 (s, 9H), 1.46- 1.45 (m, 2H), 1.30-1.18 (m, 26H), 0.80 (t, J=6.8, 3H).13C NMR (75 MHz, CDCl3- CD3OD-4:1) δ (ppm): 69.5, 67.9, 65.6, 64.4, 57.0, 52.2, 29.8, 27.6, 27.5, 27.3, 23.9, 20.6, 12.0. MS m / z (M+H): 484.5 (Calculated molecular weight-483.65). HPLC purity: 99.48%.

[0500] An analogous transformation was also successfully carried out using Compound (24) to yield Compound (12) wherein R1= C18H35D2.

[0501] When R1= C18H33D2, the following reaction scheme is employed instead. Trifluoroacetic acid (0.4 mL, 5.2 mmol) was added to a solution of Compound (24) (R1= C18H33D2) (1.65 g, 2.6 mmol) in dichloromethane (16.5 mL, 10V) at 0-5°C. The reaction mixture was allowed to stir at room temperature over a period of 1 hour. Reaction completion was monitored by TLC (80% methanol / 20% ethyl acetate / 1mL aqueous ammonia). After completion, the reaction mixture was neutralized using aqueous ammonia solution and concentrated. The crude product obtained upon evaporation of volatiles was purified through MB-20 resin column to give Compound (12) (R1= C18H33D2) as a colorless sticky solid (260 mg, 20%).1H NMR (400 MHz, CDCl3-CD3OD-4:1) δ (ppm): 5.29-5.26 (m, 2H), 4.19 (m, 2H), 3.89-3.72 (m, 3H), 3.53- 3.51 (m, 2H), 3.40-3.39 (m, 2H), 3.14 (s, 9H), 1.95-1.94 (m, 4H), 1.48 (m, 2H), 1.22- 1.20 (m, 22H), 0.81 (t, J=7.2 Hz, 3H).13C NMR (100 MHz, CDCl3-CD3OD-4:1) δ (ppm): 130.2, 130.1, 71.8, 70.2, 70.1, 67.9, 67.9, 66.8, 59.3, 54.4, 54.3, 32.2, 30.0, 29.8, 29.6, 27.5, 26.3, 22.9, 14.2. MS m / z (M+H): 510.6 (Calculated molecular weight - 509.68). HPLC purity: 98.68%. 180Ref: P329WO Example 7: Further Synthetic Routes

[0502] Other alternative syntheses starting from the allyl-protected Compound (6) are disclosed.

[0503] These syntheses are summarized in Schemes VIII and IX below, differing according to the strategy employed for attaching or elaborating the required phosphatidyl ethanolamine or phosphatidylcholine moiety starting from an allyl- protected protected alkylglycerol precursor (Compound (6)). 181Ref: P329WO182Ref: P329WO

[0504] In Scheme VIII, it is to be understood that introduction of the amine functionality at (vii-f) may also be achieved by, for example, (vii-f’) using a doubly protected amine precursor, such as by reacting HOCH2CH2NPG2 with Compound (7) to yield Compound (9b’) or by (vii-f’’) using a triply protected amine precursor, such as by reacting HOCH2CH2N+PG3 X- with Compound (7) to yield Compound (9b’’). These alternate routes are illustrated in Scheme X below. 183Ref: P329WOc e e . e a e o eso s o o pou o o pou .

[0505] In Scheme IX, it is to be understood that introduction of the amine functionality at (vii-g) may also be achieved by, for example, (vii-g’) reacting a compound HNPG2, such as diallylamine or dibenzylamine, with Compound (9a) to yield Compound (9b’), or by (vii-g’’) reacting a compound NPG3, such as triallylamine or tribenzylamine, with Compound (9a) to yield Compound (9b’’), or by (vii-g’’’) reacting an ammonia surrogate, such as an azide, such as sodium azide, with Compound (9a) to prepare Compound (9b’’’). Non-limiting examples of ammonia surrogates are disclosed in Org. Biomol. Chem., 2023, 21, 7036-7051. These ammonia surrogates are incorporated herein in entirety. These alternate routes are illustrated in Scheme XI below. 184Ref: P329WOScheme XI. Alternative Conversions of Compound (9a) to Compound (9b).

[0506] In Scheme IX, it is to be understood that introduction of the amine functionality at (vii-h2) and / or (vii-h3) may also be achieved by, for example, (vii-h2’) and / or (vii-h3’) reacting a compound HNPG2, such as dibenzylamine or diallylamine, with Compound (8b) to yield Compound (9b’), or by (vii-h2’’) and / or (vii-h3’’) reacting a compound NPG3, such as triallylamine or tribenzylamine, with Compound (8b) to yield Compound (9b’’), or by (vii-h2’’’) and / or (vii-h3’’’) reacting an ammonia surrogate, such as an azide, such as sodium azide, with Compound (8b) to prepare Compound (9b’’’). Non-limiting examples of ammonia surrogates are disclosed in Org. Biomol. Chem., 2023, 21, 7036-7051. These ammonia surrogates are incorporated herein in entirety. These alternate routes are illustrated in Scheme XII below. 185Ref: P329WOScheme XII. Alternative Conversions of Compound (9a) to Compound (9b). Example 8: Process Research and Development of 9-Step Route

[0507] The compound numbering of the present example 8 is separate to the compound numbering of the remainder of the present disclosure. Step 1

[0508] Process research and development of Step 1 was mainly focused on optimizing base and solvent. Following extensive screening procedures, the use of t- BuOK (2.0 eq.) in toluene, with heating for 2 hours was shown to provide optimum conversion (Entry 12 in Table 4-1). 186Ref: P329WOScheme 4-1. Original conditions of Step 1. Entry Conditions T Ratio LPE-11:LPE-1 (GC (h) area%)Ref: P329WO 20 t-BuOK (1.5eq.) DMF 80 2 87.4 12.6 18 907 93Step 2

[0509] The original process used 0.5 M HCl to deprotect the cyclic acetal, which carries a risk of forming potentially mutagenic impurities, e.g.4-chlorobutan-1-ol and / or dichlorobutane, from the reaction between HCl and THF. In order to minimize such risk, alternative combinations of acid / solvent were therefore investigated in a comprehensive screening study, which demonstrated that 0.1 M HCl (3 V) in methanol (10 V) heating at 60 °C over 6 hours produced the title compound in very high yield and purity (Entry 22 in Table 4-2).Entry Conditions Ratio LPE-12:LPE-11 (CAD %) Acid Solvent Temp. (oC) Time (h) LPE-12 (Product) LPE-11 (SM)Ref: P329WO 60 18 7.3 92.7 6 05 M H3PO4 (3 V) THF (10 V) 20 18 10 990Step 3

[0510] Instead of DMF / DCM, the reaction was conducted using THF (18 V), which was found to result in the formation of a clear solution. THF was therefore determined to be an improved solvent for this step both from a solubility and HSE viewpoint. 189Ref: P329WOScheme 4-3. Original conditions of Step 3. Step 4

[0511] In order to achieve better reaction performance, the following parameters were screened, including base, solvent and temperature. From the screening of the bases (Entry 1-8 in Table 4-3), t-BuOK the base used in the original conditions, remained the best choice over the other bases which were selected for review. Solvents used for the reaction were then screened which indicated that running the reaction in toluene (Entry 8-13 in Figure 4-3), results in greater conversion when compared with the reactions using the other solvents. An additional benefit of replacing THF with toluene is that the reaction can be performed with liquid-liquid extraction directly following work-up without having to initially remove the water miscible THF. Entry 9 highlighted in the table below (at 40oC), was chosen as most suitable condition for upscaling.Scheme 4-4. Original conditions of Step 4. Entry Conditions Ratio LPE-14:LPE-13:LPE-isomer:LPE-14-B1 (CAD %) Base(1.5eq.) Solvent Temp.(oC) LPE- LPE- LPE-14- LPE-14-190Ref: P329WO (1 mol / L in THF)Step 5

[0512] Due to the iron species waste generated from the reagents under the reaction conditions used in the original synthesis, adoption of other TBS deprotection conditions was investigated, e.g. formic acid (FA) and TFA in different solvents. None of these alternatives, however, resulted in a more favourable result compared with FeCl3. Subsequently, a reduction in the stoichiometry of the FeCl3 was evaluated, showing that a reduced level of this reagent was unable to achieve the 191Ref: P329WO desired performance in terms of conversion and reduced levels of other side products. The original conditions were therefore retained.Entry Conditions Ratio LPE-6 : LPE-5 : others(CAD %) Deprotection reagent Solvent Time(h) LPE-6 (Product) LPE-5(SM) othersStep 6

[0513] As Step 6 has already been optimized, the original process disclosed herein was used directly for upscaling in Example 9.Scheme 4-6. Original condition of Step 6. 192Ref: P329WO Steps 7 and 8

[0514] Due to the highly reactive nature of Compound 17, a telescoped approach has been used for Steps 7 and 8. During the present process research and development, efforts were focused mainly on the development of a purification method. In the previous reaction scheme, the crude material was purified by a hot slurry method in MeCN (60 °C), giving a waxy product with the purity of 91%. In this study, from the screening of the purification solvent system (Table 4-6), 2-MeTHF / Acetone 1:2 v / v (Entry 3 of Table 4-6) was considered to be the most suitable solvent system as it resulted in production of Compound 18 as a waxy solid, with a purity of 95.1%.. . Entry The purity of crude LPE-18 (CAD Purification Solvent (20 Temp.(°C) Purity(CAD area area %) V) %)) hand 25oC for 18h. 193Ref: P329WO Step 9

[0515] In the original reaction scheme, an expensive allyl deprotection reagent, BF3·Me2S, was used. Such reagents are unsuitable for use in upscaling activities, as the cost of this reagent accounts for approximately 40% of the total raw material costs, in addition to the environmental considerations associated with Me2S. At the beginning of the process research and development work for Step 9, the replacement of BF3·Me2S by other more suitable reagents was investigated. The screening process demonstrated that BCl3-DCM, a significantly less expensive reagent, can drive the reaction to completion. However, the positive result which was achieved was initially only possible at a small scale and it was clear that further development would be needed to use this reagent at a larger scale for this deprotection. Hence, the original reaction conditions were adopted in the current reaction scheme. Following column chromatography, the crude material still contained significant amounts of salts. A desalting process utilising reverse phase chromatography was used in the previous process. The loading for this procedure was only 500 mg / injection, which is highly inefficient and was not scalable under the conditions tested. During the current process research and development exercise, a highly efficient slurry method in MeCN:H2O=3:1 (30 V) to afford salt-free material was developed.Scheme 4-8. Original condition of Step 9. Entry Conditions Ratio LPE-(O-C18.0):LPE-18 (CAD %)194Ref: P329WO 4 BF3·Et2O(3.5eq.) + Thioanisole (3.5eq.) 18 12.3 87.7 5 BF3·2CH3COOH+ Dimethyl sulfide DCM / rt N / D N / DExample 9: Improved Production of LPE(O) compounds LPE(O)-C16.0

[0516] t-BuOK(1.5 eq.) and toluene (10 V) were added to a reactor at 25°C. The reaction temperature was cooled to 0℃. Compound 1 (1.0 eq.) and toluene (10 V) were added to the reactor at 0°C. The reaction mixture was stirred at 0°C for 1 h. 1-Bromohexadecane (1.8 eq.) and toluene (5 V) were added to the reactor. The reaction mixture was raised to 100°C and stirred for 2 h. The reaction mixture was cooled to ambient temperature then diluted with IPAc (12 V) and 5% citric acid aqueous solution (10 V). The resulting organic layers were washed with 10 wt.% NaCl (15 V) and dried over sodium sulfate, then filtered and concentrated to give an orange oil. The crude product was purified by normal-phase chromatography (n-heptane:EA, 50:1 -> 30:1). The product-containing column fractions were then combined and evaporated under reduced pressure at 40°C to provide Compound 2 (purity: 98.3% by HPLC-CAD, 80% yield) as a white waxy solid. 195Ref: P329WO

[0517] Compound 2 (1.0 equiv.) in MeOH (10 V) was added to a reactor. 0.1M aqueous HCl (3 V) was added to the reactor and the mixture was heated to 60°C for 6 h. The reaction mixture was cooled to ambient temperature. H2O (20 V) was added to the mixture and the mixture was stirred for 30 mins. After filtration, the filter cake was slurried with n-heptane / MTBE= (1:2, 6 V) overnight. Following filtration of the suspension, the filter cake was then dried to obtain Compound 3 (purity: 99% by HPLC- CAD; 88% yield) as a white solid.

[0518] Compound 3 (1 eq.) and imidazole (4 eq.) were dissolved in DCM (2 V) and DMF (10 V) at 25°C. TBSCl (1.0 eq.) was then dissolved in DMF (3 V) and the TBSCl solution was added to the reaction mixture at 25°C over 2 h. The mixture was then stirred at 30°C for 12 h.10% Aqueous NaCl solution (15 V) was added to the reaction mixture at room temperature, followed by extraction with n-heptane (15 V). The organic layer was washed with 10% aqueous NaCl solution (15 V). The organic layer was concentrated under vacuum to afford Compound 4 (crude, purity: 88% HPLC-CAD) as a light-yellow oil. This material was not purified further, as it was used directly in the next step of the synthesis. 196Ref: P329WO

[0519] Compound 4 (1.0 eq., 88%) and 3-bromoprop-1-ene (allyl bromide, 1.2 eq.) were added to toluene (10 V) at 25°C. t-BuOK (1.5 eq.) was added slowly at 25°C over 2h .The resulting reaction mixture was then stirred at 40°C for 2 h.10% aqueous NaCl solution (15 V) was added to the mixture, followed by extraction with n- heptane (15 V). The organic layer was then washed with 10% aqueous NaCl solution (15 V). The organic layer was concentrated under vacuum to afford Compound 5 (crude, purity: 79.7% by HPLC-CAD) as a light-yellow oil. This material was not purified further, as it was used directly in the next step of the synthesis.

[0520] Compound 5 (1 eq.) and FeCl3 (0.3 eq.) were added to methanol (5 V) at 25°C, then the reaction mixture was stirred at 25°C for 12 h.10% aqueous NaCl solution (10 V) was added to the mixture followed by extraction with n-heptane (10 V). The organic layer was then washed with 10% aqueous NaCl solution (10 V). The organic layer was concentrated under vacuum. The residue was purified by silica gel column chromatography (n-heptane:EA, 50:1 -> 30:1). Compound 6 (54% yield for 3 steps, purity: 98.6%) was obtained as an off-white gelatinous solid following drying under reduced pressure at 35°C.197Ref: P329WO

[0521] Compound 6 (1 eq., 98.6%) were added to a flask at 25°C. POCl3 (2 eq.) was added at 25°C in one aliquot and the reaction mixture was stirred at 25°C for 12 h. Toluene (12 V x 3) was then added and the mixture was concentrated under vacuum to give Compound 7 (IY: 98%) as a brown liquid. This material was not purified further, as it was used directly in the next step of the synthesis.

[0522] Compound 7 (1 eq.) was added to tetrahydrofuran (15 V) at 25°C and the mixture was cooled to 0°C. A solution of ethanolamine (1.5 eq.) and TEA (3 eq.) in THF (5 V) was added to the mixture at 0°C and was then stirred for 1 h at 0°C. The reaction mixture was filtered and the solids washed with THF (2 V). Aqueous HCl (2 M, 10 V) was then added to the filtrate and the mixture was stirred for 16 h at 25°C. Water (15 V) was then added to the mixture, followed by extraction with 2- methyltetrahydrofuran (10 V x 2). The organic phases were combined, washed with water (5 V x 3) and concentrated under vacuum to give crude product. The crude product was slurried with 2-methyltetrahydrofuran / acetone (1:2, 20 V) according to the following procedure: the suspension was heated at 50°C for 1 h and maintained at 25°C for 2 h. The crude product was dissolved in 2-methyltetrahydrofuran (3.5 V). Acetone (7 V) was added to the mixture dropwise. The resulting mixture was stirred at room temperature for 1 h. Following filtration and drying under reduced pressure at 35°C for 24 hours, Compound 9 was obtained as an off-white solid (purity: 97.9% by HPLC-CAD, 60% yield for 2 steps). 198Ref: P329WO

[0523] Compound 9 (1.0 eq.) was dissolved in DCM (15 V), then boron trifluoride methyl sulfide complex (4.0 eq.) was added at rt. The mixture was stirred for 16 h at rt. The reaction mixture was quenched with 5% aqueous Na2CO3 (10 V) and stirred for 30 min at rt. The pH of the mixture was then adjusted to 6-7 with 2 N aqueous HCl, the aqueous phase was separated out, then the aqueous phase was extracted with THF / 2-MeTHF (1:2, 20 V x 3). The organic phase was concentrated and then the crude product was purified by silica gel column, eluting with DCM / MeOH / H2O from 30:1:0.5 to 3:1:0.5 to give the purified product with salt. The above solid was slurried with MeCN:H2O =3:1 (30 V) at rt for 2 hours. The suspension was then filtered and the solids dried under reduced pressure at 35°C for 24 hours to obtain the product, LPE(O-C16.0) in 45% yield. LPE(O)-C18.0

[0524] t-BuOK (1.5 eq.) and toluene (10 V) were added to a reactor at 25°C. The mixture was cooled to 0℃. Compound 1 (1.0 eq.) and toluene (10.0 V) were added to the reactor at 0°C directly. The reaction mixture was stirred at 0°C for 1 h.1- bromooctadecane (1.3 equiv.) and toluene (5 V) were added to the reactor. The reaction temperature was raised to 100°C and the mixture stirred at 100°C for 2 h. The reaction mixture was cooled to ambient temperature then diluted with IPAc (12 V) and 5% citric acid aqueous solution (10 V). The resulting organic layers washed with 10 wt.% NaCl (15 V). and dried over sodium sulfate, then filtered and concentrated to give an orange oil. The crude product was purified by normal-phase chromatography (n- 199Ref: P329WO heptane:EA, 50:1 -> 30:1). The product-containing column fractions were then combined and evaporated under reduced pressure at 40°C to provide Compound 11 (purity: 98% by HPLC-CAD, 83% yield), as a white waxy solid.

[0525] Compound 11 (1 eq.) in MeOH (10 V) was added to a reactor. 0.1M aqueous HCl (3 V) was added to the reactor contents directly and the mixture was heated to 60°C and stirred at 60°C for 6 h. The reaction mixture then was cooled to ambient temperature. H2O (20 V) was added to the mixture and the reactor contents were stirred at ambient temperature for 30 mins. The resulting suspension was filtered and the filter cake was slurried with n-heptane / MTBE (1:2, 6 V) overnight and then filtered. The filter cake was dried under vacuum at 40°C for 6 hours to obtain Compound 12 (purity: 99% by HPLC-CAD; 85% yield) as a white solid

[0526] Compound 12 (1 eq.) and imidazole (4 eq.) were dissolved in DCM (2 V) and DMF (10 V) at 25°C. TBSCl (1.0 eq.) was then dissolved in DMF (3 V) and the TBSCl solution was added to the reaction mixture at 25°C over 2 h. The mixture was then stirred at 30°C for 12 h.10% Aqueous NaCl solution (15 V) was added to the reaction mixture at room temperature, followed by extraction with n-heptane (15 V). The organic layer was washed with 10% aqueous NaCl solution (15 V). The organic layer was then concentrated under vacuum to afford Compound 13 (crude, purity: 88% by HPLC-CAD) as a light-yellow oil. This material was not purified further, as it was used directly in the next step of the synthesis. 200Ref: P329WO

[0527] Compound 13 (1 eq., 88%) and 3-bromoprop-1-ene (allyl bromide, 1.2 eq.) were added to toluene (10 V) at 25°C. t-BuOK (1.5 eq.) was added slowly at 25°C over 2 h.. The resulting reaction mixture was then stirred at 40°C for 2 h.10% aqueous NaCl solution (15 V) was added to the mixture, followed by extraction with n- heptane (15 V). The organic layer was then washed with 10% aqueous NaCl solution (15 V). The organic layer was concentrated under vacuum to afford Compound 14 (crude, purity: 75% by HPLC-CAD) as a light-yellow oil. This material was not purified further, as it was used directly in the next step of the synthesis.

[0528] Compound 14 (1 eq., 75%) and FeCl3 (0.3 eq.) were added to methanol (5 V) at 25°C, then the reaction mixture was stirred at 40°C for 12 h.10% aqueous NaCl solution (10 V) was added to the mixture followed by extraction with n- heptane (10 V). The organic layer was then washed with 10% aqueous NaCl solution (10 V). The organic layer was concentrated under vacuum. The residue was purified by silica gel column chromatography (n-heptane:EA, 50:1 -> 30:1). Compound 15 (50% yield for 3 steps, purity: 98.6% by HPLC-CAD) was obtained as an off-white gelatinous solid.

[0529] Compound 15 (1 eq., 98.6%) and toluene (0.5 V) were added to a flask at 25°C. POCl3 (2 eq.) was added at 25°C in one portion and the reaction mixture 201Ref: P329WO was stirred at 25°C for 12 h. Toluene (12 V x 3) was then added and the mixture was concentrated under vacuum to give Compound 16 (IY: 97%) as a brown liquid. This material was not purified further, as it was used directly in the next step of the synthesis.p q. y V) at 25°C and the mixture was cooled to 0°C. Ethanolamine (1.5 eq.) was added to the mixture at 0°C dropwise. TEA (3 eq.) was then added to the mixture at 0°C dropwise, and THF (5 V) was added to rinse the setup. The mixture was then stirred for 1 h at 0°C. The reaction mixture was filtered and the solids washed with THF (2 V). Aqueous HCl (2 M, 5 V) was then added to the filtrate and the mixture was stirred for 16 h at 25°C. Water (15 V) was then added to the mixture, followed by extraction with 2- methyltetrahydrofuran (10 V x 2). The organic phases were combined, washed with water (5 V x 3) and concentrated under vacuum to give crude product. The crude product was dissolved in 2-methyltetrahydrofuran (3.5 V) at 50°C. The mixture was cooled down to room temperature and acetone (7 V) was added to the mixture dropwise. The resulting suspension was stirred at room temperature for 1 h. Following filtration and drying of the solids in a vacuum oven at 35°C for 24 h, Compound 18 was obtained as an off-white solid (62% yield for 2 steps, purity: 96% by HPLC-CAD).

[0531] Compound 18 (1 eq.) was dissolved in DCM (15 V), then boron trifluoride methyl sulfide complex (4.0 eq.) was added at rt and the mixture was stirred 202Ref: P329WO for 16 h at rt. The reaction mixture was quenched with 5% aqueous Na2CO3 (10 V) and stirred for 30 min at rt. The pH of the mixture was then adjusted to 6-7 with 2 N aqueous HCl, the aqueous phase was separated and then extracted with THF / 2-MeTHF (1:2, 20 V x 3). The organic phase was concentrated and then the crude product was purified by silica gel column chromatography, eluting with DCM / MeOH / H2O from 30:1:0.5 to 3:1:0.5 to give the purified product with salt. The above solid was slurried with MeCN:H2O = 3:1 (30 V) at rt for 2 hours. The suspension was then filtered and the solids were dried under reduced pressure at 35°C for 24 hours to obtain the product, LPE(O-C18.0) in 45% yield. Example 10: Alternative Synthetic Approaches Toward LPE(O) and LPC(O) Targets Synthesis of precursors - Synthesis of Strategy 1 precursor1 eq.) was dissolved in toluene (10mL, 0.5V) at 25°C. POCl3 (17.2g, 2 eq.) was added at 25°C in one aliquot and the reaction mixture was stirred at 25°C for 12 h. Toluene (240mL x 2, 12 V x 2) was then added and the mixture was concentrated under vacuum to give the C18.0 analogue of Compound 7 (24g, IY: 92%) as a light-yellow liquid. This material was stored in a refrigerator at 4°C as a stock solution in superdry THF (480mL, 50mg / mL). - Synthesis of Strategy 2 precursor:203Ref: P329WO

[0533] 2-Bromoethyl phosphorodichloridate (1.5 eq., prepared from bromoethanol and POCl3, see: Journal of Organic Chemistry 2007, 72, 8267 - 8279) was dissolved in toluene (10V) at 0°C. The C18.0 analogue of Compound 6 (1 eq., 99.1%) was dissolved in toluene (5V) and added to the mixture at 0°C. Et3N (3.0 eq.) was then added at 0°C and the reaction mixture was warmed to 25°C and stirred for 4 h. Water (5V) was added to quench the reaction, the organic and aqueous phases were separated, and the organic phase was stirred with water (5V) for 18h. The organic and aqueous phases were again separated, and the organic phase was concentrated under vacuum to afford the C18.0 analogue of Compound 9a as the desired product (26g, IY 87%). - Synthesis of Strategy 3 precursor

[0534] Triethylamine (0.79 g, 3 eq.) and ethylene chlorophosphate (0.55g, 1.5 eq.) were added to a solution of the C18.0 analogue of Compound 6 (1g, 1 eq.) in THF (10mL, 10V) at 0°C, and the reaction mixture stirred for 2h at 0°C. The resulting reaction mixture was quenched with 10% NaCl solution (10mL, 10V). The mixture was extracted with ethyl acetate (2x 10mL, 2x 10V) and the organic phases were combined. The combined organic phases were then dried over anhydrous Na2SO4 and concentrated under vacuum to give the crude C18.0 analogue of Compound 8b (1.2g, crude, IY 94%). The product is unstable, therefore it should be prepared freshly as required and used in the next step immediately. 204Ref: P329WO Strategy 1

[0535] Synthesis of a phosphorodichloridate ester, Compound 7, from Compound 6 and phosphoryl chloride as demonstrated in (vi-b), followed by reaction with an N-protected 2-aminoethanol reagent and subsequent deprotection.[ ] epresena ve expermena proce ure or e reac on o Compound 7 with a protected ethanolamine reagent, exemplified by the reaction of the C18.0 analogue of Compound 7 with BOC-protected ethanolamine: 205Ref: P329WO

[0537] 2-(N-tert-Butoxycarbonylamno)e ano ( mg, . eq.) an 3N (84 μL, 3 eq.) were added to a solution of the C18.0 analogue of Compound 7 (2 mL, 50mg / mL in THF, 1eq.) at 0°C. The reaction mixture was stirred for 1h, sampled for HPLC analysis, then allowed to warm to 25°C. The reaction was stirred for an additional 1h, sampled for HPLC analysis, then warmed to 40°C. The reaction was stirred for an additional 1h, sampled for HPLC analysis, then warmed to 70 °C. Finally, the reaction was stirred for an additional 1h, and sampled for HPLC analysis. Analytical data is tabulated below, noting that the molecular ions seen derive from reaction of the initially-formed chloro-compound with methanol during sample preparation for HPLC analysis, in addition to a disubstituted byproduct being observed in the specific case of this BOC-protected reagent.

[0538] This procedure is indicative, and full information regarding any variations to this approach is given in more detail in the table below. Numbers1 Reagent (Eq2) Base Solvent Temp Time Con-Product4Notes version C206Ref: P329WO 2-(N-tert- 25 1 82.4 8.295 8.4% butoxycarbonylamino)ethanol 71.5 product 6 0 z2: Stoichiometry with respect to Compound 7 3: Volumes with respect to mass of Compound 7 4: HPLC analysis of reaction mixture 207Ref: P329WO 5. Proposed structure is shown below. The methyl ester is formed due to the reaction mixture being quenched with methanol prior to LCMS analysis.6. Proposed structure is shown below, considered to be a doubly substituted byproduct, partial loss of BOC is assumed to occur under MS conditions.208Ref: P329WO Strategy 2

[0539] Addition of a preformed 2-haloethyl phosphorodichloridate to Compound 6 to form the corresponding 2-haloethyl phosphate ester of Compound 6, i.e. Compound (9a) followed by displacement of halide with ammonia, a protected amine, an ammonia equivalent or ammonia surrogate and subsequent deprotection.

[0050] epresentatve expermenta procedure or t e reacton o a haloethyl phosphate ester of the C18.0 analogue of Compound 6 with ammonia, a protected amine or an ammonia equivalent or ammonia surrogate, exemplified by the reaction of the C18.0 analogue of Compound 9a with 4-methoxybenzylamine: 209Ref: P329WO- e o y e ya e g, . eq. a μ , eq. were added to a solution of the C18.0 analogue of Compound 9a (100mg, 1 eq.) in THF (1 mL, 10V) at 0°C. The reaction was stirred for 1h, sampled for HPLC analysis, then allowed to warm to 25°C. The reaction was stirred for an additional 1h, sampled for HPLC analysis, then warmed to 40°C. The reaction was stirred for an additional 1h, sampled for HPLC, then warmed to 70°C. The reaction was stirred for an additional 1h, and sampled for HPLC analysis. Finally, the reaction was stirred for an additional 20h at 70 °C, and sampled for HPLC analysis. Analytical data is tabulated below.

[0542] This procedure is indicative, and full information regarding any variations to this approach is given in the table below. Numbering1Reagent Eq2Base Solvent Temp Time Conversion Product4ct ct 3210Ref: P329WO Et3N THF 25 1 0% Desired product 5.0 10 not detected 3 3 ct 4211Ref: P329WO 70 1 28.2 6.658 17.2 3 ct ct 4 ct ct 4Ref: P329WO 40 1 70.4 6.617 1.6 o 32: Stoichiometry with respect to Compound 9a 3: Volumes with respect to mass of Compound 9a 4: HPLC analysis of reaction mixture 5: Me3N used as proof of concept based on previous LPC(O) synthesis 213Ref: P329WO Strategy 3

[0543] Addition of 2-chloro-1,3,2-dioxaphospholane 2-oxide to Compound 6 to form the corresponding cyclic phosphate ester, i.e. Compound (8b) followed by ring-opening with ammonia, a protected amine, an ammonia equivalent or ammonia surrogate and subsequent deprotection.

[0544] Representative experimental procedure for the reaction of Compound 8b with ammonia, a protected amine, an ammonia equivalent or surrogate, exemplified by the synthesis of the C18.0 analogue of Compound 9b from 4- methoxybenzylamine. 214Ref: P329WO- y y g, . q. μ , q.) was added to the solution of the C18.0 analogue of Compound 8b (100 mg, 1 eq.) in DMSO (1 mL, 10V) at 0°C. The reaction was stirred for 1h, sampled for HPLC analysis, then allowed to warm to 25°C. The reaction was stirred for an additional 1h, sampled for HPLC analysis, then warmed to 40°C. The reaction was stirred for an additional 1h, sampled for HPLC analysis, then warmed to 70°C. The reaction was stirred for an additional 1h, and sampled for HPLC analysis. The reaction was stirred for an additional 20h at 70°C, and sampled for HPLC analysis.

[0546] This procedure is indicative, and full information regarding any variations to this approach is given in in the table below. Patent numbering1Reagent Eq2Base Solvent T Time Conversion Product4and / orRef: P329WO 20 Et3N DMSO rt, 24 72.6% 5.0 10 40,216Ref: P329WO 40 1 14.0 6.574 2.92: Stoichiometry with respect to Compound 8b 3: Volumes with respect to mass of Compound 8b 217Ref: P329WO 4: HPLC analysis of reaction mixture 5: Me3N used as proof of concept based on previous LPC(O) synthesis 218Ref: P329WO Removal of O-allyl and N-allyl protection (BOC, trityl and allyl) from Compound 9b as a penultimate or final step

[0547] The deprotection of three Compound 9b variants (C18.0: N-BOC, N-trityl and N-allyl ) was investigated to determine the feasibility of removal of these protecting groups, with or without simultaneous O-allyl deprotection. The substrates for these deprotection studies were prepared as described previously and isolated using a simplified work up without chromatographic purification. The crude products were then subjected to the deprotection conditions directly. A representative example of this workup is shown below for the C18.0 BOC-protected version of Compound 9b.

[0548] 2-(N-tert-Butoxycarbonylamino)ethanol (480 mg, 1.5 eq.) and Et3N (840 μL, 3 eq.) were added to the stock solution of the C18.0 analogue of Compound 7 (20 mL, 50mg / mL in THF, 1eq.) at 0°C. The reaction was warmed to 25°C and stirred for 16h. Water (5mL, 5V) was then added to quench the reaction. The mixture was extracted with ethyl acetate (2x 10mL, 2x 10V) and the organic phases were combined. The combined organic phases were dried over sodium sulphate and concentrated under vacuum to give the crude C18.0 analogue of Compound 9b. Strategy Compound 9b precursor (C18.0 analogue) rotectin rou s StructureRef: P329WO 9b Substrate HPLC profile Deprotection Temp Time Product3m / z (C18.0 of substrate1Conditions21.2. Reactions conducted at 100mg scale (substrate) 3. HPLC analysis of reaction mixture 4. The observed mass corresponds to triphenylmethylium ion, shown below, which is produced by fragmentation of the substrate. Under different MS conditions the molecular ion can also be observed as shown in the table above describing Strategy 1 investigations.220Ref: P329WO 5. The two mono-deprotected products are not distinguishable by MS at this point, their identities are assigned based on retention time. 6. Monodeprotected product only. Example 11: Proposed Synthesis of Cyclic Prodrugs of LPE(O)

[0549] Method A: Compound 8 (1 eq.) is dissolved in DCM (15 V), then boron trifluoride methyl sulfide complex (4.0 eq.) is added at rt. The mixture is stirred for 16 h at rt. The reaction mixture is quenched with 5% aqueous Na2CO3 (10 V) and is stirred for 30 min at rt. It is expected that analysis of the crude reaction mixture by LC MS reveals formation of the title product.

[0550] Method B: Dimethylbarbituric acid (1.2 equiv.) is added to Compound (8) in dry methanol (12 mL) and the resulting mixture is sparged with argon for 25 minutes. Palladium (0) tetrakis(triphenylphosphine) (0.06 equiv.) is added in one portion and the reaction mixture is heated. It is expected that analysis of the crude reaction mixture by LC MS reveals formation of the title product.221

Claims

Ref: P329WO CLAIMS 1. A method for synthesizing Compound (6) , or a salt thereof, the method comprising:(i) reacting (R)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1-OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2);(ii) hydrolyzing Compound (2) in a mixture comprising acid to form Compound (3);(iii) reacting Compound (3) with a protecting group reagent in solution, the protecting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4), wherein PG is a protecting group;222Ref: P329WO (iv) reacting Compound (4) in a mixture comprising an allylating reagent to form Compound (5);and (v) deprotecting Compound (5) in solution to form Compound (6).

2. A method for synthesizing Compound (10) , or a salt thereof, the method(i) reacting (R)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanol (Compound (1)) with one of R1-OSO2R2and R1-X, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group, R2is a C1-6 alkyl group or a C6-18 aryl group, and X is Cl, Br, or I, in solution to form Compound (2);223Ref: P329WO (ii) hydrolyzing Compound (2) in a mixture comprising acid to form Compound (3);(iii) reacting Compound (3) with a protecting group reagent in solution, the protecting group reagent reacting with the primary alcohol of Compound (3) to form Compound (4), wherein PG is a protecting group;(iv) reacting Compound (4) in a mixture comprising an allylating reagent to form Compound (5);(v) deprotecting Compound (5) in solution to form Compound (6);(vi) phosphorylating the primary alcohol of Compound (6) to form Compound (7); 224Ref: P329WO(vii) reacting Compound (7) with ethanolamine in solution to form Compound (8);(viii) reacting Compound (8) in a mixture comprising acid to form Compound (9);and (ix) deprotecting Compound (9) to form Compound (10) .225Ref: P329WO 3. The method of claim 1, wherein compound (6) is further reacted to form Compoun a salt thereof, the metho(vi-a) reacting Compound (6) with 2-bromoethyl phosphorodichloridate to form Compound (9a);(vii-a) reacting Compound (9a) in a solution comprising a metal oxide and trimethylamine to form Compound (11);and (viii-a) deprotecting Compound (11) to form Compound (12) .226Ref: P329WO 4. The method of claim 1, wherein Compound (6) is further reacted to form Compoun a salt thereof, the metho(vi-b) phosphorylating the primary alcohol of Compound (6) to form Compound (7);(vii-b) reacting Compound (7) with ethanolamine in solution to form Compound (8);(viii-b) reacting Compound (8) in a mixture comprising an acid to form Compound (9);(ix-b) methylating compound (9) to form Compound (11); 227Ref: P329WO and(x-b) deprotecting Compound 11 to form Compound (12) .

5. The method of claim 1, wherein compound (6) is further reacted to form Compound (10) , or a salt thereof, the method comp(vi-c) reacting bis(2-cyanoethyl)-N,N-diisopropylphosphoramidite with Fmoc- protected aminoethanol to give the corresponding diisopropylphosphoramidite Compound (16) (16)228Ref: P329WO and reacting Compound (16) with Compound (6) to form Compound (13);(vii-c) reacting Compound (13) with an oxidizing reagent to form Compound (14)(viii-c) selectively deprotecting Compound (14) at the secondary alcohol to form Compound (15); d(ix-c) deprotecting Compound (15) to form Compound (10) .229Ref: P329WO 6. The method of any one of claims 1 to 5, wherein (i) comprises reacting Compound (1) and the R1-OSO2R2or the R1-X in the presence of a base.

7. The method of any one of claims 1 to 6, wherein the base used in (i) is an alkali metal hydroxide, an alkali metal alkoxide, or alkali metal hydride, such as sodium hydroxide, potassium hydroxide, sodium tert-butoxide, potassium tert-butoxide, sodium hydride, or potassium hydride.

8. The method of claim 7, wherein the base is potassium tert-butoxide.

9. The method of any of claims 6-8, wherein from 1 to 4 equivalents of the base is used, such as from 1 to 3, such as about 2 equivalents.

10. The method of any of claims 6-9, wherein (i) is conducted in an apolar solvent or a polar solvent, for example wherein (i) is conducted in toluene.

11. The method of any of claims 6-10, wherein (i) comprises reacting Compound (1) for at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, such as at least 2 hours, for example from 15 minutes to 48 hours.

12. The method of any one of claims 1 to 10, wherein (i) comprises heating the solution.

13. The method of any one of claims 1 to 12, wherein (i) comprises heating the solution at a temperature range of about 60°C to 100°C.

14. The method of claim 13, wherein (i) comprises heating the solution at a temperature of about 80°C.

15. The method of any one of claims 1 to 7, wherein (i) comprises reacting Compound (1) with the R1-OSO2R2or R1-X at room temperature.

16. The method of any of claims 1 to 7, wherein (i) comprises reacting Compound (1) with R1- OSO2R2. 230Ref: P329WO 17. The method of any of claims 1 to 13, wherein (i) comprises reacting Compound (1) with R1- X, for example wherein X is bromide, and optionally wherein R1is a C10-C24 alkyl or C10-C24 alkenyl.

18. The method of any one of claims 1 to 17, wherein (i) comprises adding tetrabutylammonium bromide to the solution.

19. The method of claim 18, wherein the addition of tetrabutylammonium bromide occurs after the addition of R1-OSO2R2or R1-X.

20. The method of any one of claims 1 to 19, wherein (i) further comprises purifying Compound (2) before hydrolyzing Compound (2).

21. The method of any one of claims 1 to 19, wherein, in (i), Compound (2) is not purified before Compound (2) is hydrolyzed.

22. The method of any one of claims 1 to 21, wherein (ii) comprises dissolving Compound (2) in a solvent before hydrolyzing Compound (2).

23. The method of claim 22, wherein the solvent is a polar aprotic solvent, a polar protic solvent, or a mixture thereof.

24. The method of any of claims 22-23, wherein the solvent is selected from the group consisting of: tetrahydrofuran, diethyl ether, and 2-methyl tetrahydrofuran, for example tetrahydrofuran.

25. The method of any of claims 22-23, wherein the solvent is a polar protic solvent, for example methanol.

26. The method of any of claims 22-25, wherein the solution is heated at from 50 °C to 70 °C, such as about 60 °C.

27. The method of any of the preceding claims, wherein (ii) comprises reacting Compound (2) for at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, such as at least 2 hours, for example from 15 minutes to 48 hours. 231Ref: P329WO 28. The method of any one of claims 1 to 23, wherein the acid in (ii) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate.

29. The method of any one of claims 1 to 28, wherein (ii) comprises heating the mixture to 50°C to 80°C.

30. The method of claim 29, wherein (ii) comprises heating the mixture to 65°C.

31. The method of any one of claims 1 to 30, wherein (ii) comprises extracting Compound (3) from the mixture with a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example 2-methyltetrahydrofuran.

32. The method of any of one of claims 1 to 31, wherein (ii) comprises reacting with HCl in methanol at for example from 50 °C to 70 °C, such as 60 °C over the course of for example from 4 hours to 18 hours, such as 6 hours.

33. The method of any of one of claims 1 to 32, wherein (ii) comprises reacting with from 0.05M to 2.00 M HCl, such as from 0.1M to 1.50M, for example from 0.1M to 0.5M HCl, optionally using from 1 to 10 Volumes, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 Volumes of HCl.

34. The method of any one of claims 1 to 33, wherein, in (iii), the solution comprises one or more of pyridine, dichloromethane, imidazole, and dimethylformamide.

35. The method of any one of claims 1 to 34, wherein, in (iii), the temperature of the solution is maintained at from -10 °C to 40 °C, such as from about 0°C to room temperature, for example between about 25 and 30 °C.

36. The method of any one of claims 1 to 35, wherein the protecting group reagent in (iii) is selected from tert-butyldimethylsilyl chloride, trimethylsilyl chloride, triethylsilyl chloride, triisopropylsilyl chloride,tert-butyl diphenyl silyl chloride, a benzyl halide, and a triphenylmethyl halide. 232Ref: P329WO 37. The method of any of claims 1-36, wherein, in (iii), the solution comprises a polar solvent, such as a polar aprotic or polar protic solvent.

38. The method of claim 37, wherein the solution comprises an ethereal solvent, for example THF, 2-MeTHF, or diethyl ether, for example THF.

39. The method of claim 36, wherein the protecting group reagent in (iii) is tert- butyldimethylsilyl chloride.

40. The method of any of claims 1-39, wherein the protecting group reagent in (iii) is added in a stoichiometry of from 0.9 to 3.0 equivalents, such as from 1.0 to 2.5 equivalents, for example from 1.0 to 2.0 equivalents, relative to the Compound (2).

41. The method of any of claims 1-40, wherein, in (iii), a base, such as a nucleophilic base is added to the solution, for example a base capable of electrophilic activation of the protecting group reagent.

42. The method of claim 41, wherein the base is selected from the group consisting of: imidazole, pyridine, 4-DMAP, triethylamine, N-methylimidazone, and 2,6-lutidine.

43. The method of any of claims 41-42, wherein from 2 to 8 equivalents of the base is added to the solution relative to Compound (2), such as from 2 to 7 equivalents, such as from 3 to 6 equivalents, for example from 4 to 5 equivalents.

44. The method of any one of claims 1 to 43, wherein the allylating reagent in (iv) is an allyl halide, such as allyl bromide or allyl chloride.

45. The method of claim 44, wherein the allyl halide is allyl bromide.

46. The method of any one of claims 1 to 45, wherein, in (iv), the mixture comprises essentially dry tetrahydrofuran, for example having a water content determined by Karl Fisher of from 0.0 ppm to 80 ppm, such as from 0.0 ppm to 70 ppm, such as from 0.0 ppm to 60 ppm, such as from 0.9 ppm to 50 ppm. 233Ref: P329WO 47. The method of any of claims 1 to 46, wherein, in (iv), the mixture comprises an apolar solvent, for example a water immiscible solvent, such as toluene.

48. The method of any of claims 1 to 46, wherein, in (iv), the mixture comprises toluene, THF, MTBE, DMAc, n-heptane, and / or acetonitrile.

49. The method of any one of claims 1 to 48, wherein, in (iv), the mixture comprises a non-nucleophilic base, for example a tert-butoxide.

50. The method of any one of claims 1 to 49, wherein, in (iv), the mixture comprises a base which is a metal hydride and / or a metal alkoxide.

51. The method of claim 49, wherein the metal hydride is sodium hydride and / or the metal alkoxide is potassium tert-butoxide.

52. The method of any one of claims 1-51, wherein (iv) comprises cooling the mixture at a temperature of from -10 °C to 50 °C, such as from about 0°C to room temperature, for example between about 25 °C to about 45 °C.

53. The method of any of claims 1-52, wherein, in (iv), from 1.0 to 2.5 equivalents of the base is used, such as from 1.1 to 2.0 equivalents, for example from 1.2 to 1.8 equivalents, for example about 1.5 equivalents.

54. The method of any of claims 1-53, wherein, in (iv), from 1.0 to 2.5 equivalents of the allylating reagent is used, such as from 1.1 to 2.0 equivalents, for example from 1.2 to 1.8 equivalents, for example about 1.5 equivalents.

55. The method of any one of claims 1 to 54, wherein (v) comprises selectively deprotecting the primary alcohol of Compound (5) to form Compound (6).

56. The method of any one of claims 1 to 55, wherein, in (v), the solution of Compound (5) comprises tetrahydrofuran and a fluoride ion source.

57. The method of claim 56, wherein the fluoride ion source is tetrabutylammonium fluoride (TBAF). 234Ref: P329WO 58. The method of any of claims 1 to 57, wherein, in (v), a Lewis acid is added to the mixture, for example a Lewis acid selected from the group consisting of: FeCl3, TiCl4, BCl3. ZnBr₂, ZnCl₂, AlCl₃, BF₃·Et₂O, Sc(OTf)₃, InCl₃, and Cu(OTf)₂ 59. The method of any one of claims 1 to 58, wherein Compound (6) is selected from d60. The method of any one of claims 2 and 6 to 59 wherein (vi) comprises reacting Compound (6) with a phosphoryl halide to form Compound (7).

61. The method of claim 60, wherein the phosphoryl halide is phosphoryl trichloride.

62. The method of any one of claims 2 and 6 to 61, wherein (vi) comprises reacting Compound (6) with a phosphoryl halide at a temperature of from -20 °C to 30 °C, such as from -10 °C to 10 °C for example about 0°C under an inert atmosphere.

63. The method of any one of claims 2 and 6 to 62, wherein in (vi), from 1.0 to 3.0 equivalents of the phosphoryl halide is used, such as from 1.2 equivalents to 2.8 equivalents, such as from 1.4 equivalents to 2.6 equivalents, for example from 1.6 equivalents to 2.4 equivalents, for example about 2.0 equivalents.

64. The method of any one of claims 2 and 6 to 63, wherein (vi) comprises reacting Compound (6) in a apolar solvent, such as toluene, optionally using from 0.2 volumes to 5 volumes of toluene relative to Compound (6).

65. The method of any one of claims 2 and 6 to 64, wherein (vi) comprises reacting Compound (6) over the course of from 4 hours to 48 hours, such as from 6 hours to 42 hours, such as from 8 hours to 36 hours, such as from 10 hours to 30 hours, for example from 12 hours to 22 hours, for example about 16 hours. 235Ref: P329WO 66. The method of any one of claims 2 and 6 to 65, wherein (vi) comprises adding a solution of Compound (6) to a solution comprising phosphoryl halide dropwise.

67. The method of any one of claims 2 and 6 to 66, wherein (vi) comprises dissolving Compound (6) in a solution comprising triethylamine and diethyl ether.

68. The method of any one of claims 2 and 6 to 67, wherein (vii) comprises dissolving Compound (7) in a solution comprising a polar aprotic solvent, for example diethyl ether, tetrahydrofuran, 2-MeTHF, or a mixture thereof.

69. The method of claim 68, wherein the solution comprises 2-aminoethanol and an amine base, such as triethylamine.

70. The method of claim 69, wherein from 1.0 to 3.0 equivalents of 2-aminoethanol is used, such as from 1.2 equivalents to 2.8 equivalents, such as from 1.4 equivalents to 2.6 equivalents, for example about 1.5 equivalents.

71. The method of claim 70, wherein from 1.5 to 4.0 equivalents of the amine base is used, such as from 1.8 equivalents to 3.7 equivalents, such as from 2.0 equivalents to 3.5 equivalents, for example about 3.0 equivalents, optionally wherein the amine base is triethylamine.

72. The method of any of claims 2 and 6 to 71, wherein in (vii), the temperature is from -20 °C to 30 °C, such as from -15 °C to 25 °C, for example from -10 °C to 20 °C, optionally from -10 °C to 10 °C, for example about 0 °C.

73. The method of any of claims 2 and 6 to 72, wherein in (vii), Compound (7) is reacted for from 5 minutes to 24 hours, such as from 10 minutes to 14 hours, such as from 15 minutes to 8 hours, such as from 30 minutes to 6 hours, such as from 45 minutes to 4 hours, for example about 1 hour.

74. The method of any one of claims 2 and 6 to 73, wherein (vii) comprises heating the solution at a temperature of about 30°C to 60°C under an inert atmosphere. 236Ref: P329WO 75. The method of claim 74, wherein (vii) comprises heating the solution at a temperature of about 50°C with an internal temperature of 40°C.

76. The method of any one of claims 2 and 6 to 75, wherein the acid in (viii) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate.

77. The method of any one of claims 2 and 6 to 76, wherein, in (viii), the mixture comprises diethyl ether and tetrahydrofuran.

78. The method of any one of claims 2 and 6 to 76, wherein (viii) comprises adding an acid, such as an aqueous acid, for example aqueous HCl to the mixture.

79. The method of any one of claims 2 and 6 to 78, wherein the solution and any precipitate therein in (vii) subsequent to mixing of 2-aminoethanol and Compound (7) is subjected to filtration to obtain a filtrate used directly in step (viii).

80. The method of any one of claims 78 to 79, wherein step (viii) further comprises a purification step subsequent to the addition of the acid, wherein the purification step comprises: a. subjecting a composition of Compound (9) to a solvent system, b. heating the solvent system to a predefined temperature for a predefined period of time, c. allowing the solvent system to cool to a temperature lower than the predefined temperature thereby obtaining a composition of Compound (9) having increased purity relative to the composition of Compound (9) in step a.

81. The method of claim 80, wherein the predefined temperature is from 30 °C to 100 °C, such as from 35 °C to 90 °C, such as from 40 °C to 80 °C, such as from 45 °C to 70 °C, for example from 50 °C to 65 °C, for example about 60 °C. 237Ref: P329WO 82. The method of any of claims 80-81, wherein the predefined period of time is at least 15 minutes, such as at least 30 minutes, for example from 15 minutes to 48 hours, for example about 1 hour.

83. The method of any of claims 80-82, wherein the solvent system comprises a polar aprotic solvent or a mixture of different polar aprotic solvents.

84. The method of any of claims 80-83, wherein the solvent system comprises a solvent selected from the group consisting of: MeCN, MTBE, 2-MeTHF, acetone, IPAc, and a mixture thereof.

85. The method of claim 84, wherein the solvent system comprises 2-MeTHF and acetone, for example in a mixture of 2-MeTHF and acetone with from 10% to 50% 2- MeTHF, the balance being acetone, for example about 1:2 v / v 2-MeTHF:acetone.

86. The method of any of claims 80-85, wherein the crude composition of Compound (9) is added to a first solvent, such as an ethereal solvent, for example 2-MeTHF, after which a second solvent is added to the first solvent, for example acetone.

87. The method of any one of claims 2 and 6 to 86, wherein (ix) comprises reacting Compound (9) with at least one catalyst and under an inert atmosphere.

88. The method of claim 87, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing.

89. The method of any one of claims 2 and 6 to 88, wherein (ix) comprises reacting Compound (9) in a mixture that is heated at a temperature of about 40°C to 70°C.

90. The method of claim 89, wherein the temperature is about 50°C to 55°C.

91. The method of any of claims 2 and 6 to 87, wherein (ix) comprises reacting Compound (9) with a Lewis acid. 238Ref: P329WO 92. The method of claim 91, wherein the Lewis acid comprises boron, such as a boron halide.

93. The method of any of claims 91-92, wherein the Lewis acid is selected from the group consisting of: BF3, BCl3, and BBr3.

94. The method of any of claims 91-93, wherein the Lewis acid is selected from the group consisting of: BF3·Et2O. and BCl3·DCM, for example wherein the Lewis acid is BCl3·DCM.

95. The method of any of claims 91-94, wherein (ix) comprises adding a ligand to the Lewis acid, for example wherein the ligand is a sulfide-based ligand, such as dimethylsulfide, thioanisole, and thiosalicylic acid, for example wherein the ligand is dimethylsulfide.

96. The method of any of claims 91-95, wherein from 1.5 to 6.0 equivalents of the Lewis acid is used in (ix), such as from 2.0 equivalents to 4.5 equivalents, such as from 2.5 equivalents to 4.0 equivalents, for example about 3.5 equivalents.

97. The method of any of claims 91-96, wherein from 1.5 to 6.0 equivalents of the ligand is used in (ix), such as from 2.0 equivalents to 4.5 equivalents, such as from 2.5 equivalents to 4.0 equivalents, for example about 3.5 equivalents or about 4.0 equivalents.

98. The method of claim 97, wherein from 2.0 equivalents to 4.5 equivalents of the Lewis acid and ligand is used, for example about 3.5 equivalents for the Lewis acid and the ligand or about 4.0 equivalents.

99. The method of any of claims 87-98, wherein (ix) is conducted in dichloromethane.

100. The method of any of claims 91-99, wherein in (ix), the reaction is conducted for at least 1 hour, such as from 1 hour to 1 week. 239Ref: P329WO 101. The method of any of claims 87-100, wherein (ix) further comprises a silane, such as a silane selected from the group consisting of: triethylsilane, phenylsilane, diphenylsilane, polymethylhydrosiloxane (PMHS), triisopropylsilane, trimethoxysilane, dimethoxymethylsilane, and tetramethyldisiloxane (TMDS).

102. The method of claim 101, wherein from 2.0 to 8.0 equivalents of the silane is used, for example from 3.0 to 7.0, such as about 6.0 equivalents.

103. The method of any of claims 87-102, wherein the temperature in (ix) is from 0 °C to 80 °C, such as from 10 °C to 70 °C, such as from 15 °C to 60 °C, for example from about 20 °C to about 50 °C, for example about 20 °C to 40 °C.

104. The method of claim 3, wherein the metal oxide in (vii-a) is silver oxide.

105. The method of claim 3 or claim 104, wherein (viii-a) comprises reacting Compound (11) with at least one catalyst and under an inert atmosphere.

106. The method of claim 105, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing.

107. The method of claim 4, wherein (vi-b) comprises reacting Compound (6) with a phosphoryl halide to form Compound (7).

108. The method of claim 107, wherein the phosphoryl halide is phosphoryl trichloride.

109. The method of any one of claims 4, 107, and 108, wherein (vi-b) comprises reacting Compound (6) with a phosphoryl halide at a temperature of about 0°C under an inert atmosphere.

110. The method of any one of claims 4 and 107 to 109, wherein (vi-b) comprises dissolving Compound (6) in a solution comprising triethylamine and diethyl ether. 240Ref: P329WO 111. The method of any one of claims 4and 107 to 110, wherein (vi-b) comprises adding a solution of Compound (6) to a solution comprising a phosphoryl halide dropwise.

112. The method of any one of claims 4 and 107 to 111, wherein (vii-b) comprises dissolving Compound (7) in a solution comprising diethyl ether and tetrahydrofuran.

113. The method of claim 112, wherein the solution comprises ethanolamine and triethylamine.

114. The method of any one of claims 4 and 107 to 113, wherein (vii-b) comprises heating the solution at a temperature of about 30°C to 60°C under an inert atmosphere.

115. The method of claim 114, wherein (vii-b) comprises heating the solution at a temperature of about 50°C with an internal temperature of 40°C.

116. The method of any one of claims 4 and 107 to 115, wherein the acid in (viii-b) is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, an alkali metal bisulfate, and an alkali metal dihydrogen phosphate.

117. The method of any one of claims 4 and 107 to 116, wherein, in (viii-b), the mixture comprises diethyl ether and tetrahydrofuran.

118. The method of any one of claims 4 and 107 to 117, wherein, in (ix-b), methylating Compound (9) comprises reacting Compound (9) with methyl bromide, methyl iodide, or dimethyl sulfate.

119. The method of any one of claims 4 and 107 to 117, wherein (x-b) comprises reacting Compound (11) with at least one catalyst and under an inert atmosphere.

120. The method of claim 119, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing. 241Ref: P329WO 121. The method of claim 5, wherein in (vii-c), the oxidising reagent is selected from a peroxy acid or a salt thereof, potassium peroxymonosulfate, and magnesium monoperoxyphthalate.

122. The method of claim 121, wherein the peroxy acid is selected from meta- chloroperoxybenzoic acid and peroxyacetic acid.

123. The method of any one of claims 5, 121, or 122, wherein (viii-c) comprises reacting Compound (14) with at least one catalyst and under an inert atmosphere.

124. The method of claim 123, wherein the at least one catalyst is selected from tetrakis(triphenylphosphine)palladium(0), tetrakis(triphenylphosphine)palladium(0), bis(dibenzylideneacetone) palladium(0), tris(dibenzylideneacetone)dipalladium(0), palladium on carbon, palladium (II) chloride, palladium (II) hydroxide on carbon, tris(triphenylphosphine)rhodium(I) chloride, and a catalyst derived from any of the foregoing or generated in situ from any of the foregoing.

125. The method of any one of claims 5 and 121 to 124, wherein (ix-c) comprises reacting Compound (15) with piperidine to form Compound (10).

126. A method for synthesizing Compound (12) , or a salt thereof, the metho(1-a) reacting R1-OH with (2R)-(-)-glycidyl tosylate (Compound (18)) to form Compound (19); 242Ref: P329WO(19), wherein R1is selected from a C3-C30 alkyl group, a C3- C30 alkenyl group, and a C3-C30 acyl group, (1-b) reacting Compound (19) with a benzylating reagent in solution, the benzylating reagent reacting with the secondary alcohol of Compound (19) to form Compound (20); (20), wherein Ar is selected from phenyl and 4- meth(1-c) reacting Compound (20) with a metal acetate to form Compound (21); );(1-e) reacting Compound (22) with ethylene chlorophosphate to form Compound (23); 3)243Ref: P329WO (1-f) reacting Compound (23) in a solution comprising trimethylamine to form Compound (24); d(1-g) deprotecting Compound (24) to form Compound (12) .

127. The method of claim 126, wherein the metal acetate is caesium acetate.

128. A method for synthesizing Compound (10) , or a salt thereof, the method comp(vi-d) phosphorylating the primary alcohol of Compound (6) to form Compound (7);244Ref: P329WO(vii-d) reacting Compound (7) with ethanolamine in solution to form Compound (8); d either[1] (viii-d1) reacting Compound (8) in a mixture comprising acid to form Compound (9)and (ix-d1) deprotecting Compound (9) to form Compound (10); or[2] (viii-d2) deprotecting Compound (8) to form Compound (8a) 245Ref: P329WO d(ix-d2) reacting Compound (8a) in a mixture comprising acid to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

129. A method for synthesizing Compound (10) , or a salt thereof, the method comp(vi-e) phosphorylating the primary alcohol of Compound (6) to form Compound (7); 246Ref: P329WO(vii-e) reacting Compound (7) with ethanolamine in solution to form Compound (9); d(viii-e) deprotecting Compound (9) to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

130. A method for synthesizing Compound (10) , or a salt thereof, the method comp247Ref: P329WO (vi-f) phosphorylating the primary alcohol of Compoun to form Compound (7);(vii-f) reacting Compound (7) with an amino-protected ethanolamine compound in solution to form Compound (9b); , wherein PG is a protecting group[1] (viii-f1) selectively deprotecting Compound (9b) in a mixture to form Compound (9) and(ix-f1) deprotecting Compound (9) to form Compound (10); 248Ref: P329WO r[2] (viii-f2) selectively deprotecting Compound (9b) in a mixture to form Compound (9c) d(ix-f2) deprotecting Compound (9c) to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

131. The method of claim 130, wherein in (vii-f), the amino-protected ethanolamine compound is selected from the group consisting of: a. 2-(N-tert-butoxycarbonylamino)ethanol, to provide Compound (9b), wherein PG is Boc; b. benzyl ethanolamine, to provide Compound (9b), wherein PG is benzyl; c. 2-[(4-methoxybenzyl)amino]ethanol, to provide Compound (9b), wherein PG is 4-methoxybenzyl; d. 2-(allylamino)ethanol, to provide Compound (9b), wherein PG is allyl; and 249Ref: P329WO e. N-tritylethanolamine, to provide Compound (9b), wherein PG is trityl.

132. The method of any of claims 130-131, wherein in (vii-f), a base is added to the solution, such as an amine base or a metal carbonate.

133. The method of claim 132, wherein the amine base is triethylamine.

134. The method of any of claims 132-133, wherein in (vii-f) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (7), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

135. The method of any of claims 132-134, wherein in (vii-f) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

136. The method of any of claims 132-135, wherein in (vii-f) the temperature is from - 10 °C to 80 °C, such as from 0 °C to 70 °C, for example from 10 °C to 60 °C.

137. The method of any of claims 132-136, wherein in (vii-f) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

138. A method for synthesizing Compound (10) 250Ref: P329WO , or a salt thereof, the method comp(vi-f) phosphorylating the primary alcohol of Compoun to form Compound (7);(vii-f) reacting a di-protected amino compound HOCH2CH2NPG2 with Compound(7) to yield Compoun , wherein eachPG is independently[1] (viii-f1’) selectively deprotecting Compound (9b’) in a mixture to form Compound (9) and251Ref: P329WO (ix-f1) deprotecting Compound (9) to form Compound (10); or[2] (viii-f2) selectively deprotecting Compound (9b) in a mixture to form Compound (9c’) and(ix-f2) deprotecting Compound (9c’) to form Compound (10) , wherein R1is a C3-C30alkyl, C3-C30139. The method of claim 138, wherein in (vii-f’), the di-protected amino compound HOCH2CH2NPG2 is N,N-dibenzyl-2-aminoethanol to provide Compound (9b’), wherein each PG is benzyl.

140. The method of any of claims 138-139, wherein in (vii-f’), a base is added to the solution, such as an amine base or a metal carbonate. 252Ref: P329WO 141. The method of claim 140, wherein the amine base is triethylamine.

142. The method of any of claims 138-141, wherein in (vii-f’) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (7), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

143. The method of any of claims 138-142, wherein in (vii-f’) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

144. The method of any of claims 138-143, wherein in (vii-f’) the temperature is from - 10 °C to 80 °C, such as from 0 °C to 70 °C, for example from 10 °C to 60 °C.

145. The method of any of claims 138-144, wherein in (vii-f’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

146. A method for synthesizing Compound (10) , or a salt thereof, the method comp253Ref: P329WO (vi-f) phosphorylating the primary alcohol of Compound (6) );(vii-f’’) reacting a compound HOCH2CH2N+PG3 X- with Compound (7) to yield Compoun , wherein each PG isindepend nion, such as Cl-, Br- - -, I, OH, or other suitable counter anion, and either [1] (viii-f1’’) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9)and (ix-f1) deprotecting Compound (9) to form Compound (10); 254Ref: P329WO r[2] (viii-f2) selectively deprotecting Compound (9b ) in a mixture to form Compound (9c’’) d(ix-f2) deprotecting Compound (9c ) to form Compound (10) , wherein R1is a C3-C30alkyl, C3-C30147. A method for synthesizing Compound (10) , or a salt thereof, the method comp255Ref: P329WO (vi-g) reacting Compound (6)with 2-bromoethyl phosphorodichloridate to form Compod (9a);(vii-g) converting Compound (9a) to Compound (9b); d either[1] (viii-g1) selectively deprotecting Compound (9b) in a mixture to form Compound (9c) d(ix-g1) deprotecting Compound (9c) to form Compound (10); 256Ref: P329WO r[2] (viii-g2) selectively deprotecting Compound (9b) in a mixture to form Compound (9) d(ix-g2) deprotecting Compound (9) to form Compound (10) or(vii-gx) converting Compound (9a) to Compound (9) ;wherein R is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group and PG is a protecting group. 257Ref: P329WO 148. The method of claim 147, wherein (vii-gx) comprises reacting Compound 9a with ammonia, for example methanolic ammonia.

149. The method of any of claims 147-148, wherein in (vii-gx) no other base than ammonia is added.

150. The method of any of claims 147-148, wherein in (vii-gx) from 1.0 equivalents to 5.0 equivalents of a base is used with respect to Compound (9a), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents, for example wherein the base is an amine base, such as triethylamine.

151. The method of any of claims 147-150, wherein in (vii-gx) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

152. The method of any of claims 147-151, wherein in (vii-gx) the temperature is from 30 °C to 100 °C, such as from 35 °C to 90 °C, for example from 40 °C to 85 °C.

153. The method of any of claims 147-152, wherein in (vii-gx) the reaction is performed over the course of at least 30 minutes, such as at least 45 minutes, such as at least 1 hour, such as from 30 minutes to 120 hours.

154. The method of any of claims 147-153, wherein in (vii-gx) from 10 to 50 equivalents of ammonia is used, such as from 10 to 40 equivalents, for example 20 equivalents. 258Ref: P329WO 155. The method of claim 147, wherein in (vii-g), Compound 9a is reacted with a protected amine (PGNH2) selected from the group consisting of: a. benzylamine to provide Compound (9b), wherein PG is benzyl; b. 4-methoxybenzyl to provide Compound (9b), wherein PG is 4- methoxybenzyl; and c. Allylamine, for example allylamine·HCl, to provide Compound (9b), wherein PG is allyl.

156. The method of any of claims 147, 155, wherein in (vii-g) from 1.0 to 5.0 equivalents of the protected amine is used, such as from 1.2 equivalents to 5.0 equivalents, for example from 1.5 equivalents to 4.0 equivalents, such as about 3.0 equivalents or such as about 4.0 equivalents.

157. The method of any of claims 147, 155-156, wherein in (vii-g) a base is used, such as an amine base.

158. The method of claim 157, wherein the amine base is triethylamine.

159. The method of any of claims 147, 155-158, wherein in (vii-g) Compound (9a) is reacted with the protected amine in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

160. The method of any of claims 147, 155-159, wherein in (vii-g) the temperature is from 10 °C to 90 °C, such as from 15 °C to 85 °C, for example from 20 °C to 80 °C, such as about 70 °C. 259Ref: P329WO 161. The method of any of claims 147, 155-160, wherein in (vii-g) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

162. A method for synthesizing Compound (10) , or a salt thereof, the method comp(vi-g) reacting Compound (6) with 2-bromoethyl phosphorodichloridate to form Compound (9a);(vii-g) converting Compound (9a) to Compound (9b') by reacting a compound HNPG2 with Compound (9a) to yield Compound (9b’) 260Ref: P329WO , wherein each PG is independently a[1] (viii-g1) selectively deprotecting Compound (9b’) in a mixture to form Compound (9c’)and (ix-g1) deprotecting Compound (9c’) to form Compound (10); or[2] (viii-g2) selectively deprotecting Compound (9b) in a mixture to form Compound (9) and261Ref: P329WO (ix-g2) deprotecting Compound (9) to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

163. The method of claim 162, wherein in (vii-g’), Compound 9a is reacted with a di- protected amine (PG2NH), optionally wherein the di-protected amine is dibenzylamine.

164. The method of any of claims 162, 163, wherein in (vii-g’) from 1.0 to 5.0 equivalents of the protected amine is used, such as from 1.2 equivalents to 5.0 equivalents, for example from 1.5 equivalents to 4.0 equivalents, such as about 3.0 equivalents or such as about 4.0 equivalents.

165. The method of any of claims 162, 163-164, wherein in (vii-g’) a base is used, such as an amine base.

166. The method of claim 165, wherein the amine base is triethylamine.

167. The method of any of claims 162, 163-166, wherein in (vii-g’) Compound (9a) is reacted with the di-protected amine in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether. 262Ref: P329WO 168. The method of any of claims 162, 163-167, wherein in (vii-g’) the temperature is from 10 °C to 90 °C, such as from 15 °C to 85 °C, for example from 20 °C to 80 °C, such as about 70 °C.

169. The method of any of claims 162, 155-168, wherein in (vii-g’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

170. A method for synthesizing Compound (10) , or a salt thereof, the method comp(vi-g) reacting Compound (6)with 2-bromoethyl phosphorodichloridate to form Compound (9a);(vii-g ) converting Compound (9a) to Compound (9b'’) by reacting a compound NPG3 with Compound (9a) to yield Compound (9b’’) 263Ref: P329WO , wherein each PG is independently a protecting[1] (viii-g1) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9c’’) d(ix-g1) deprotecting Compound (9c ) to form Compound (10); r[2] (viii-g2) selectively deprotecting Compound (9b ) in a mixture to form Compound (9)and (ix-g2) deprotecting Compound (9) to form Compound (10) 264Ref: P329WO ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

171. The method of claim 170, wherein in (vii-g’’), Compound 9a is reacted with a tri- protected amine (PG3N), optionally wherein the tri-protected amine is trimethylamine.

172. The method of any of claims 170-171, wherein in (vii-g’’) a metal oxide catalyst is added, such as wherein the metal oxide catalyst is silver oxide, for example wherein from 0.5 to 5.0 equivalents of the metal oxide catalyst is used, such as from 0.6 equivalents to 5.0 equivalents, for example from 0.7 equivalents to 4.0 equivalents, such as about 1.0 equivalents.

173. The method of any of claims 170-172, wherein in (vii-g’’) Compound (9a) is reacted with the tri-protected amine in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

174. The method of any of claims 170-173, wherein in (vii-g’’) the temperature is from 10 °C to 90 °C, such as from 15 °C to 85 °C, for example from 20 °C to 80 °C, such as about 70 °C.

175. The method of any of claims 170-174, wherein in (vii-g’’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours. 265Ref: P329WO 176. A method for synthesizing Compound (10) , or a salt thereof, the method comp(vi-g) reacting Compound (6)with 2-bromoethyl phosphorodichloridate to form Compound (9a);(vii-g ) converting Compound (9a) to Compound (9b'’’) by reacting Compound (9a) with a metal azide to yield Compound (9b’’’) d either[1] (viii-g1) selectively deprotecting Compound (9b’’’) in a mixture to form Compound (9c’’’) 266Ref: P329WO O N3and (ix-g1) reducing Compound (9c’’’) to form Compound (10); r[2] (viii-g2) reducing Compound (9b ) to form Compound (9) d(ix-g2) deprotecting Compound (9) to form Compound (10) , wherein R1is a C3-C30alkyl, C3-C3267Ref: P329WO 177. The method of claim 176, wherein the metal azide in (vii-g’’’) is selected from the group consisting of: sodium azide, and potassium azide.

178. The method of any of claims 176-177, wherein in (vii-g’’’) from 1.0 to 10.0 equivalents of the metal azide is used, such as from 1.5 to 9.0 equivalents, such as from 2.0 to 8.0 equivalents, such as from 3.0 to 7.0 equivalents, for example about from 4.0 to 6.0 equivalents, for example about 5.0 equivalents.

179. The method of any of claims 176-178, wherein in (vii-g’’’) Compound (9a) is reacted with the metal azide in a polar aprotic solvent, such as an ethereal polar aprotic solvent, for example wherein the ethereal polar aprotic solvent is selected from the group consisting of: THF, 2-MeTHF, and diethyl ether.

180. The method of any of claims 176-179, wherein in (vii-g’’’) the temperature is from 0 °C to 100 °C, such as from 15 °C to 90 °C, for example from 20 °C to 80 °C.

181. The method of any of claims 176-180, wherein in (vii-g’’’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

182. A method for synthesizing Compound (10) , or a salt thereof, the method comp268Ref: P329WO (vi-h) reacting Compound (6)with ethylene glycol chlorophosphate to form Compound (8b); r[1] (vii-h1) converting Compound (8b) to Compound (9) d(viii-h1) deprotecting Compound (9) to form Compound (10); or[2] (vii-h2) converting Compound (8b) to Compound (9b), 269Ref: P329WO(viii-h2) selectively deprotecting Compound (9b) in a mixture to form Compound (9c), d(ix-h2) deprotecting Compound (9c) to form Compound (10); or[3] (vii-h3) converting Compound (8b) to Compound (9b),(viii-h3) selectively deprotecting Compound (9b) to form Compound (9) 270Ref: P329WOand (ix-h3) deprotecting Compound (9) in a mixture to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group and PG is a protecting group.

183. The method of claim 182, wherein (vii-h1) comprises reacting Compound (8b) with ammonia or an ammonium halide, such as ammonium chloride or methanolic ammonia.

184. The method of claim 182, wherein in (vii-h2) or (vii-h3), Compound (8b) is reacted with an amine selected from the group consisting of: a. benzylamine, to provide Compound (9b), wherein PG is benzyl; b. 4-methoxybenzylamine, to provide Compound (9b), wherein PG is 4- methoxybenzyl; and c. allylamine, to provide Compound (9b), wherein PG is allyl.

185. The method of claim 184, wherein at least 1.0 equivalent of the amine is added, such as from 1.0 to 5.0 equivalents, such as from 1.1 equivalents to 4.5 equivalents, for example from 1.5 equivalents to 4.0 equivalents, for example about 3.0 equivalents. 271Ref: P329WO 186. The method of any of claims 182-185, wherein in (vii-h1), (vii-h2) or (vii-h3), a base is added to the solution, such as an amine base or a metal carbonate.

187. The method of claim 186, wherein the amine base is triethylamine.

188. The method of any of claims 182-187, wherein in (vii-h1), (vii-h2) or (vii-h3) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (8b), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

189. The method of any of claims 182-188, wherein in (vii-h1), (vii-h2) or (vii-h3) the solution comprises a polar aprotic solvent, such as an ethereal polar aprotic solvent or a chlorinated solvent, for example wherein the polar aprotic solvent is selected from the group consisting of: THF, DCM, 2-MeTHF, DMSO, DMF, MeCN, diethyl ether, and a mixture thereof.

190. The method of any of claims 182-189, wherein in (vii-h1), (vii-h2) or (vii-h3) the temperature is from 0 °C to 120 °C, such as from 10 °C to 110 °C, for example from 20 °C to 100 °C.

191. The method of any of claims 182-190, wherein in (vii-h1), (vii-h2) or (vii-h3) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

192. A method for synthesizing Compound (10) 272Ref: P329WO , or a salt thereof, the method comp(vi-h) reacting Compound (6)with ethylene glycol chlorophosphate to form Compound (8b); er[1] (vii-h1) converting Compound (8b) to Compound (9)and (viii-h1) deprotecting Compound (9) to form Compound (10); 273Ref: P329WO or[2] (vii-h2) reacting a compound HNPG2 with Compound (8b) to yield Compound , wherein each PG is independently(viii-h2’) selectively deprotecting Compound (9b’) in a mixture to form Compound (9c’), d(ix-h2) deprotecting Compound (9c) to form Compound (10); or274Ref: P329WO [3] (vii-h3’) reacting a compound HNPG2 with Compound (8b) to yield Compound(9b’) , wherein each PG is independently a protecting; (viii-h3) selectively deprotecting Compound (9b’) to form Compound (9) d(ix-h3) deprotecting Compound (9) in a mixture to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

193. The method of claim 192, wherein in (vii-h2’) or (vii-h3’), Compound (8b) is reacted with dibenzylamine, to provide Compound (9b’), wherein each PG is benzyl.

194. The method of claim 193, wherein at least 1.0 equivalent of dibenzylamine is added, such as from 1.0 to 5.0 equivalents, such as from 1.1 equivalents to 4.5 275Ref: P329WO equivalents, for example from 1.5 equivalents to 4.0 equivalents, for example about 1.1 equivalents.

195. The method of any of claims 192-194, wherein in (vii-h2’) or (vii-h3’), a base is added to the solution, such as an amine base or a metal carbonate.

196. The method of claim 195, wherein the amine base is triethylamine.

197. The method of any of claims 192-196, wherein in (vii-h2’) or (vii-h3’) from 1.0 equivalents to 5.0 equivalents of the base is used with respect to Compound (8b), such as from 1.5 equivalents to 4.5 equivalents, for example from 2.0 equivalents to 4.0 equivalents, for example about 3.0 equivalents.

198. The method of any of claims 192-197, wherein in (vii-h2’) or (vii-h3’) the solution comprises a polar aprotic solvent, for example wherein the polar aprotic solvent is selected from the group consisting of: DMSO, and DMF, and a mixture thereof.

199. The method of any of claims 192-198, wherein in (vii-h2’) or (vii-h3’) the temperature is from 0 °C to 120 °C, such as from 10 °C to 110 °C, for example from 20 °C to 100 °C.

200. The method of any of claims 192-199, wherein in (vii-h2’) or (vii-h3’) the reaction is performed over the course of at least 10 minutes, such as at least 15 minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 48 hours.

201. A method for synthesizing Compound (10) 276Ref: P329WO , or a salt thereof, the method comp(vi-h) reacting Compound (6)with ethylene glycol chlorophosphate to form Compound (8b); er[1] (vii-h1) converting Compound (8b) to Compound (9)and (viii-h1) deprotecting Compound (9) to form Compound (10); 277Ref: P329WO , or[2] (vii-h2’’) reacting a compound NPG3 with Compound (8b) to yield Compound ,wherein each PG is independently a(viii-h2’’) selectively deprotecting Compound (9b’’) in a mixture to form Compound (9c’’), d(ix-h2) deprotecting Compound (9c ) to form Compound (10); r278Ref: P329WO [3] (vii-h3’’) reacting a compound NPG3 with Compound (8b) to yield Compound(9b, wherein each PG is independently aprotecting group; (viii-h3) selectively deprotecting Compound (9b’’) to form Compound (9)and (ix-h3) deprotecting Compound (9) in a mixture to form Compound (10) ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group and PG is a protecting group.

202. A method for synthesizing Compound (10) , or a salt thereof, the method comp279Ref: P329WO (vi-h) reacting Compound (6)with ethylene glycol chlorophosphate to form Compound (8b); r[1] (vii-h1) converting Compound (8b) to Compound (9) d(viii-h1) deprotecting Compound (9) to form Compound (10); or[2] (vii-h2 ) reacting Compound (8b) with a metal azide to yield Compound (9b’’’); 280Ref: P329WO (viii-h2’’’) selectively deprotecting Compound (9b’’’) in a mixture to form Compound (9c’’’), O 3and (ix-h2) reducing Compound (9c’’’) to form Compound (10); or[3] (vii-h3 ) reacting Compound (8b) with a metal azide to yield Compound (9b’’’) ;(viii-h3) reducing Compound (9b’’’) to form Compound (9)and (ix-h3) deprotecting Compound (9) in a mixture to form Compound (10) 281Ref: P329WO ,wherein R1is a C3-C30alkyl, C3-C30alkenyl, or C3-C30acyl group.

203. The method of claim 202, wherein the metal azide in (vii-h2’’’) or (vii-h3’’’) is selected from the group consisting of: sodium azide, and potassium azide.

204. The method of any of claims 202-203, wherein in (vii-h2’’’) or (vii-h3’’’) from 1.0 to 10.0 equivalents of the metal azide is used, such as from 1.5 to 9.0 equivalents, such as from 2.0 to 8.0 equivalents, such as from 3.0 to 7.0 equivalents, for example about from 4.0 to 6.0 equivalents, for example about 5.0 equivalents.

205. The method of any of claims 202-204, wherein in (vii-h2’’’) or (vii-h3’’’) Compound (8b) is reacted with the metal azide in a solvent which is: a polar aprotic solvent, a polar protic solvent, or a combination thereof, for example wherein the solvent is selected from the group consisting of: H2O, THF, 2-MeTHF, acetone, and a mixture thereof; for example wherein the solvent comprises H2O and acetone in a from 1:5 to 1:2 mixture, such as in a 1:3 mixture.

206. The method of any of claims 202-205, wherein in (vii-h2’’’) or (vii-h3’’’) the temperature is from 0 °C to 100 °C, such as from 15 °C to 90 °C, for example from 20 °C to 80 °C.

207. The method of any of claims 202-206, wherein in (vii-h2’’’) or (vii-h3’’’) the reaction is performed over the course of at least 10 minutes, such as at least 15 282Ref: P329WO minutes, such as at least 30 minutes, such as at least 1 hour, for example from 15 minutes to 120 hours.

208. The method of any one of claims 1 to 207, wherein R1is a C10-C24 alkyl, such as C15-C20 alkyl, for example a C16-C18 alkyl, C10-C24 alkenyl, such as a C15-C20 alkenyl, for example a C16-C18 alkenyl, or a C10-C24 acyl, such as a C15-C20 acyl, for example a C16- C18 acyl group.

209. The method of any one of claims 1 to 208, wherein R1is a C16 alkyl group.

210. The method of any one of claims 1 to 208, wherein R1is a C18 alkyl group.

211. The method of any one of claims 1 to 208, wherein R1is a C18 alkenyl group.

212. The method of any one of claims 2, 5-125, and 128-211, wherein Compound (10) ,283Ref: P329WO 213. The method of any one of claims 2, 4, 126, and 127, wherein Compound (12) is ,214. A method for synthesizing Compound (25) , or a salt thereof, the method comp(2-a) providing Compound (26) , wherein each of R1and R3is independently selected from-C30 alkenyl group, and a C3-C30 acyl group. 284Ref: P329WO (2-b) phosphorylating the primary alcohol of Compound (26) to form Compound (27);(2-c) reacting Compound (27) with ethanolamine in solution to form Compound (28); d(2-d) reacting Compound (28) in a mixture comprising acid to form Compound (25) .

215. The method of claim 214, wherein R1is selected from a C16-C18 alkyl group, a C16-C18 alkenyl group, and a C16-C18 acyl group.

216. The method of claim 214, wherein R3is an allyl group.

217. The method of claim 216, further comprising removing the allyl group to form Compound (10) 285Ref: P329WO , wherein R1is a C3-C30 alkyl, C3-C218. The method of claim 214, wherein Compound (25) is further reacted to form Comp(3-a) methylating Compound (25) to form Compound (29); d(3-b) deprotecting Compound (25) to form Compound (12) , wherein R1is a C3-C30 alkyl, C3-C3286Ref: P329WO 219. A compound selected from: Compoun ,Compoun ,Compoun , Compoun ,Compoun ,Compoun ,287Ref: P329WO , , , ,ependently selected from a C16-C18 alkyl group, a C16-C18 alkenyl group, and a C16-C18 acyl group.

220. The compound of claim 219, wherein R1is a C16 alkyl group.

221. The compound of claim 219, wherein R1is a C18 alkyl group.

222. The compound of claim 219, wherein R1is a C18 alkenyl group.

223. A compound selected from 288Ref: P329WO (R)-4-((hexadecyloxy)methyl)-2,2-dimethyl-1,3-dioxolane ;; ;-oxide ;289Ref: P329WO 2-((R)-2-(allyloxy)-3-(octadecyloxy)propoxy)-1,3,2-oxazaphospholidine 2-oxide ;lidine 2- oxid ;;;;290Ref: P329WO phos; ; 291Ref: P329WO ;and salts thereof.

224. The compound or salt thereof of claim 223, wherein the compound or salt is not marine-sourced.

225. A composition comprising one or more compounds or salts thereof of any one of claims 219 to 224.

226. The composition according to claim 225, wherein the compound or salt thereof is not marine-sourced.

227. The compound or salt thereof of any one of claims 219 to 224, wherein the compound or salt is produced by the method of any one of claims 1 to 218.

228. A method of using at least one compound selected fromCompound (1 ,292Ref: P329WO ;wherein the method comprises subjecting Compound (10), Compound (12), Compound (9), and / or Compound (11) to one or more of the following transformations: (A) dehydrogenation of R1, and / or (B) acylation of the secondary alcohol to incorporate an acyl group, wherein R1is selected from a C3-C30 alkyl group, a C3-C30 alkenyl group, and a C3-C30 acyl group.

229. The method of claim 228, wherein Compound (10) is subjected to transformation (A) to yield Compoun , wherein R5is selectedacyl group. 293Ref: P329WO 230. The method of claim 228, wherein Compound (10) is subjected to transformation (B) to yield Compoun , wherein R1is selected C3-C30acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group.

231. The method of claim 228, wherein Compound (10) is subjected to transformation (A) and transformation (B) to yield Compound (10AB) , wherein R5is selected from28 acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group.

232. The method of claim 228, wherein Compound (12) is subjected to transformation (A) to yield Compound (12A) , wherein R5is selected from8 acyl group. 294Ref: P329WO 233. The method of claim 228, wherein Compound (12) is subjected to transformation (B) to yield Compoun , wherein R1is selected C3-C30acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group.

234. The method of claim 228, wherein Compound (12) is subjected to transformation (A) and transformation (B) to yield Compound (12AB) , wherein R5is selected from8 acyl group and R6is selected from a C1-C30 alkyl group and a C1-C30 alkenyl group.

235. The method of claim 228, wherein Compound (9) is subjected to transformation (A) to yield Compound (9A , wherein R5is selected from28 acyl group. 295Ref: P329WO 236. The method of claim 228, wherein Compound (11) is subjected to transformation (A) to yield Compoun , wherein R5is selected 1-C28acyl group.

237. The method of any one of claims 228-231, wherein compound (10) is formed from the method of any one of claims 2, 5-73, and 76-92.

238. The method of any one of claims 228 and 232-234, wherein compound (12) is formed from the method of any one of claims 3, 4, 126, and 127. 296