Efficient synthesis of β-L-5-[(E)-2-bromovinyl)-1-((2S,4S)-2-(hydroxymethyl)-1,3-(dioxolane-4-yl)uracil)](L-BHDU) via chiral pure L-dioxolane

Abstract

The present invention relates to a novel method for synthesizing L-BDHU and related starting materials and intermediate compounds from (7-J-2,2-dimethyl-1,3-dioxolane-4-carboxylate (7)) in at least 7 steps. The present invention eliminates the expensive chiral purification of L-dioxolane (11) to produce enantiomerically concentrated (ee > 99%) compounds. Optically pure compound 11 was used to produce L-BHDU (15) and its monophosphate prodrug (POM-L-BHDU, 22). The novel synthesis method and the novel intermediate compounds represent further embodiments of the present invention.

Description

Technology Field

[0001] The present invention ( R) -2,2-dimethyl-1,3-dioxolane-4-carboxylate( 7 The present invention relates to a novel method for synthesizing L-BDHU and related starting materials and intermediate compounds from ) in at least 7 steps. The present invention (L-dioxolane ( 11 By excluding the expensive chiral purification of ), it produces an enantiomerically enriched (ee > 99%) compound. Optically pure compound 11 L-BHDU( 15 ) and its monophosphate prodrug (POM-L-BHDU, 22 It was used to manufacture ). A novel synthesis method and a novel intermediate compound represent additional embodiments of the present invention. Background Technology

[0002] Related applications

[0003] This application claims the benefit of priority of U.S. provisional application serial number us63 / 621,722 filed January 17, 2024 and us63 / 716,343 filed November 5, 2024, the entire contents of each application are incorporated by reference into this application in their entirety.

[0004] Background and Overview of the Invention

[0005] The varicella zoster virus (VZV) is a highly contagious alpha herpesvirus that causes chickenpox (varicella) upon initial infection and causes shingles (herpes zoster) upon reactivation from the latent period. 1 The U.S. Centers for Disease Control and Prevention (CDC) estimated that there are approximately 1 million cases of shingles in the United States every year. 2People over 50 years of age, immunocompromised individuals, people infected with human immunodeficiency virus (HIV), and organ transplant patients are at a higher risk of VZV infection. 3 VZV infection leads to serious complications and causes a painful, blistering skin rash containing an abundance of infectious viruses. A major problem with shingles is postherpetic neuralgia, which is prolonged pain that persists for months or years even after the rash has healed. 4 Therefore, effective and safe novel antiviral agents are needed for the treatment and management of VZV infection. However, while vaccination is available for both stages of VZV infection and an adjuvant subunit vaccine is approved for the prevention of herpes zoster, these can only be used in healthy individuals. 5

[0006] Acyclovir (ACV), valacyclovir (VACV), and famciclovir (FCV) Fig. 1 ) is an approved drug and is used to treat VZV infection. 6 However, these approved drugs do not show high efficacy against VZV and require high doses and long-term use, which typically promotes drug resistance. 7 Cidofovir (CDV), a broad-spectrum antiviral, is active against VZV, but it is associated with nephrotoxicity and cannot be administered orally. 6 Another antiviral drug, brivudine (BVDU), is approved in Europe for the treatment of VZV infection. 8, 9 A major disadvantage associated with BVDU is that it is metabolized into bromovinyl-uracil (BVU) in the liver. 8BVU inhibits the activity of dihydropyridine dehydrogenase (DPD), which oxidizes thymidine and uracil. Therefore, there are life-threatening concerns regarding the combination therapy with brivudine in cancer patients being treated with 5-fluorouracil (5-FU). 10 Due to the serious adverse effects of currently approved drugs, there is an urgent need for new antiviral agents to treat VZV and its resistant strains.

[0007] Over the past 20 years, in the search for superior antiviral compounds against VZV, herpes simplex viruses 1 and 2, and Epstein-Barr virus (EBV), the inventors' research group has been involved in the discovery of modified D and L-dioxolane-nucleos(t)ides. In these efforts, the inventors have identified 2-(hydroxymethyl)-1,3-dioxolane-4-yl]5-vinyluracil (L-HDVD, ) which possesses potent antiviral activity against EBV, VZV, and herpes simplex virus type 1 (HSV1). Fig. 2 ) 11 2-Bromovinyl-2-(hydroxymethyl)-1,3-dioxolane uracil (L-BHDU) 12, 13 Invented. L-HDVD is EBV (EC 50 (value 0.01 μM) and Kaposi's sarcoma-associated herpesvirus (KSHV) (EC 50 It exhibits high activity for (= 0.09 μM). 11 L-BHDU has a selectivity index (SI) >909 and exhibits no cytotoxicity in human foreskin fibroblasts (HFF) up to 200 μM EC 50 The value was proven to be 0.25 μM. 12 In vivo In the study, L-BHDU significantly reduces viral load compared to ACV and VACV. It is noteworthy that metabolic studies found that L-BHDU does not inhibit DPD and has a superior safety and antiviral profile compared to brivudine. 12

[0008] Recently, in order to increase the cellular bioavailability and absorption of L-BHDU with enhanced pharmacokinetic properties, the inventors developed an L-BHDU monophosphate pivaloyloxymethyl ester prodrug (POM-L-BHDU-MP, Fig. 2 Developed ). 14, 15 POM-L-BHDU-MP exhibited antiviral activity similar to L-BHDU in infected cells. In contrast, POM-L-BHDU-MP was superior to L-BHDU in a VZV mouse model. 16 Pharmacokinetic (PK) studies revealed that POM-L-BHDU-MP had superior oral absorption compared to L-BHDU. 14 A recent PK study also revealed that POM-L-BHDU-MP releases nearly identical concentrations of L-BHDU into the blood via the IV route or oral administration. This demonstrates that POM-L-BHDU-MP exhibits equivalent efficacy when administered orally compared to IV administration. Repeat analysis in humanized mice showed that POM-L-BHDU-MP was effective when administered orally once daily at doses of 11.3 mg / kg or higher, indicating its efficacy and bioavailability in vivo represents. 16 POM-L-BHDU-MP was evaluated as a topical treatment for VZV and HSV infections in a human skin exfoliation model and was highly effective against both viruses when formulated at 0.2% in cocoa butter. 16 Based on these findings, POM-L-BHDU-MP was selected for preclinical development against VZV and HSV-1 infections.

[0009] To develop POM-L-BHDU-MP as a preclinical candidate, additional extended biological, pharmacokinetic, and toxicological studies are required, which necessitates a significant amount of POM-L-BHDU-MP. Therefore, the development of a robust, practical, and cost-effective synthesis procedure for POM-L-BHDU-MP was necessary. The prior synthesis of L-BHDU involved the L-dioxolane core intermediate ( 5, Fig. 3, Reaction Equation 1 It was performed by Sznaidman et al. 17, 18 The procedure reported in this article is L-dioxane 5 It was used in the synthesis of, and is an important intermediate for the synthesis of L-modified nucleoside analogs.

[0010] Reaction Equation 1 . Racemidioxolane( 4 Synthesis of chiral pure D and L dioxolanes through chiral separation of ). 17

[0011]

[0012] Reagents and Conditions: (a) DMAP, Et3N, tert -Butylmethyl ether, room temperature; (b) BF3.Et2O, acetonitrile 0°C-room temperature.

[0013] However, this reported process is racemicioxolane ( 4 It involved chiral chromatographic separation of ), which limits the scalability and commercial utility of Scheme 1. In addition, during large-scale synthesis, the intermediate in the presence of an excess of boron trifluoride diethyl etherate (BF3.Et2O) 4 The decomposition of was observed. Chiral pure D and L-dioxolane ( 5 and 6 In addition, it revealed stability issues during the chiral separation process, and unwanted impurities were formed during the concentration or freeze-drying of the collected chiral pure fraction. Reaction Equation 1These disadvantages limit the process for large-scale synthesis of both L and D dioxolanes (5 and 6). To overcome the problem of Reaction Scheme 1, the inventors herein [describe] methyl ( R )-2,2-dimethyl-1,3-dioxolane-4-carboxylate, 7 We report the scalable and practical synthesis of L-BHDU in seven steps. However, the process reported in this document is a revised procedure, specifically intended for the synthesis of L-dioxolane previously reported by Bera S. et al. 19 The previously reported procedures and patents contain a great deal of unclear information regarding the specific synthesis of chiral pure L-dioxolane (enantiomeric excess ≥ 99%) and have posed challenges to medicinal chemists. 20 Nevertheless, the present application relates to the direct synthesis and purification of chiral pure L-dioxolane through the formation of diastereoisomer chiral amine salts. This novel procedure also excludes chiral separation and specific steps of column chromatography, which makes the process suitable for the scalable synthesis of POM-L-BHDU-MP for clinical and commercial applications. Brief explanation of the drawing

[0014] Fig. 1 This shows the chemical structures of several antiviral drugs for the treatment of VZV infection. Fig. 2 This shows the chemical structures of L-HDVD, L-BHDU, and POM-L-BHDU-MP. Fig. 3, Reaction Equation 1 racemidioxolane ( 4 Illuminates the chemical synthesis of chiral pure D and L dioxolanes through chiral separation of ). 17 Reagents and conditions : (a) DMAP, Et3N, tert -Butylmethyl ether, room temperature; (b) BF 3. ET2O, Acetonitrile 0℃-room temperature. Fig. 4, Reaction Equation 2 is the chiral pure important intermediate L-dioxolane ( 11The synthesis of L-BHDU through ) is illustrated. Reagents and Conditions: (a) p -TSA or Dowex 50W X8 toluene at 80°C; (b) 1 M aqueous LiOH, THF, 0°C-room temperature; (c) i) ( S (ii) Et-phenylethylamine, acetonitrile 75°C-room temperature; BF3.Et2O, acetonitrile 0°C-room temperature; (ii) EtOAc:IPA (3:1), room temperature; (d) 1 N aqueous HCl, room temperature; (e) Manganese diacetate (Mn(II)(OAc)2), diacetoxy iodobenzene, acetic acid, 2-Me-THF 65°C, or alternatively Pb(OAc)4, pyridine, acetonitrile, 0°C-room temperature; (f) Bromovinyl uracil, TBDMSOTf, TMSI, DCM, room temperature; (g) BCl3, DCM, -78°C. Fig. 5 is (a) compound 10 (b) shows that the ORTEP diagram confirms the relative stereochemistry of L-dioxolane. (b) shows the stereostructures of D-dioxolane and L-dioxolane; in the case of D-dioxolane, the C-2 and C-4 hydrogens are cis, whereas in L-dioxolane, the C-2 and C-4 hydrogen atoms are trans to each other. Fig. 6, Reaction Equation 3 is the revised L-BHDU Synthesis of L-dioxolane 11 intermediate from 18 The city is created through. Reagents and Conditions: (a) 5% Pd / C, H2 gas, MeOH, room temperature; (b) Isobutyric anhydride, pyridine, DMAP, DCM, 0°C to room temperature; (c) Mn(OAc)2, diacetoxy iodobenzene, acetic acid, 2-Me-THF, 65°C, or alternatively Pb(OAc)4, pyridine, acetonitrile, 0°C to room temperature; (d) Bromovinyl-uracil, TBDMSOTf, TMSI, DCM, room temperature; (e) 7 N NH3 solution in methanol, room temperature. Fig. 7, Reaction Equation 4 is the synthesis of the POM-L-BHDU-MP prodrug L-BHDU( 15) It is made through. Reagents and Conditions: (a) NMI, THF, 0°C to room temperature, 3 hours. Specific details for implementing the invention

[0015] Brief description of the invention

[0016] In embodiments, the present invention relates to L-BDHU through a chemical precursor and a chemical intermediate ( 15 This relates to a method for synthesizing ), thereby enabling the following efficient synthesis.

[0017] L-BHDU ( 15 ), POM-L-BHDU-MP ( 22 ) and a number of precursors and / or chemical intermediates used in the synthesis of these compounds.

[0018] In one embodiment, the present invention relates to a compound (according to the synthesis step described in Reaction Scheme 2 of the attached FIG. 4) 7 L-BDHU from ) 15 This concerns a method for synthesizing ). Compound 7 It first proceeds in four steps to the pure core intermediate L-dioxolanic acid ( 11 It is converted to ), and subsequently to L-dioxolanic acid 11 L-BHDU by one of the two synthesis routes disclosed herein ( 15 It is converted to ). According to the method of the first embodiment, methyl-( R -)-2,2-dimethyl-1,3-dioxolane-4-carboxylate(compound 7 ) is first reacted with 2-(benzyloxy)acetaldehyde in a solvent (preferably toluene) with a catalytic amount of acid (preferably toluene / p-toluenesulfonic acid or Dowex 50W X8) at an elevated temperature (above room temperature, e.g., 60-100°C for several hours) to produce a benzyl-protected dioxolane ester compound as described herein. 8 L-dioxolane compounds by providing a mixture of diastereomers (often 1:1) of the compound in high yield (often >70%) 11 It is used to generate. Subsequent compounds 8 It undergoes ester hydrolysis in an aqueous solvent / base (e.g., 1 M LiOH / THF) to remove the ester group and the corresponding benzyl-protected carboxylate compound. 9 It provides, commonly in yields exceeding 90%. Compound 9 through the formation of acid-base salts ( S )-Complexing with phenylethylamine to substantially pure (i.e., de > 90%, commonly de > 99%) compound 10 It provides, first, a benzyl-protected racemic carboxylate compound 9 ( S )-Complexes with phenylethylamine to precipitate and / or crystallize acid amine 10 A mixture of salts of the D and L isomers (compounds 10M A compound that provides (referred to as ). 10 Provides a salt of, said salt is a mixture of both D and L isomers (15% D, 85% L, de > 70%, compound mixture 10M (referred to as), comprising said mixture, said mixture was dissolved (taken up) (preferably several times) in a 3:1 volume ratio of ethyl acetate:isopropyl alcohol, stirred and precipitated at room temperature for several hours (e.g., 3, 4, or 5, often 4 hours). said mixture was dissolved 2 to 5 times, often 3 to 5 times in preferred embodiments, often 3 times to obtain substantially pure ( de > 90%, commonly > 99% ( S )-phenylethylammonium salt (acid amine salt) 10 provided. After (S )-phenylethylammonium acid amine salt 10 Dissolve in methylene chloride (DCM), expose to a cold aqueous acid (e.g., 1 N aqueous HCl solution), sequentially wash with brine, and dry (e.g., sodium sulfate) to obtain the compound 11(L-dioxolane) A compound that is diastereomerically pure or substantially diastereomerically pure ( de > 90%, de > 95%, commonly > It provides 99%, most commonly 99+%-100%). Therefore, the compound 11 (Trans / L (as a form) is a compound by precipitation without chromatographic separation. 9 It is obtained in high purity from. Note that one or more of the aforementioned steps are performed in a separate step or in a single-pot reaction vessel.

[0019]

[0020]

[0021] In one embodiment, a compound 11 The compound as described below in this specification and in the attached FIG. 4 and Reaction Scheme 2 12 and 13 through compounds 15 It is converted to. In this embodiment, the compound 11 The benzyl-protected dioxolane acetate compound is first reacted at 65°C with Mn(II)OAc, diacetoxy iodobenzene, acetic acid, and 2-Me-THF (or alternatively with lead tetraacetate Pb(OAc)4 in acetonitrile / base, commonly with pyridine or trimethylamine, more commonly with pyridine), the components are combined at 0°C and then heated at room temperature for 16 hours, steps not shown in the reaction scheme below), and subjected to chromatographic separation (preferably column chromatography, eluted with ethyl acetate / hexane). 12 of αand β Anomeric mixture ( α / β The ratio is commonly about 2:1. compound 12 ( α and β As a mixture of anomers) in a dry solvent (e.g., dichloromethane (DCM)). tert - After being coupled to bromovinyl uracil using butyldimethylsilyl trifluoromethanesulfonate (TBDMSOTf), iodotrimethylsilane (TMSI), and 2,4,6-collidine, and then chromatographed (e.g., silica gel column, 1:4 EtOAc / hexane as eluent), α - Isome compounds 14 small amount of component (about 10-20% yield, not desired) and desired β - Isome compounds 13 second > It is provided with a 50% yield (often 55% or more). Subsequent compounds 13 The compound undergoes deprotection of the benzyl group (e.g., using BCl3 in DCM at a low temperature, e.g., -78°C). 15 cast > It is provided at a yield of 50%. Each of the aforementioned synthesis steps can be performed as a separate step or together in a single reaction vessel.

[0022]

[0023]

[0024] In one embodiment, the present invention relates to L-dioxolanic acid as described below in this specification and in the attached FIG. 6 and Reaction Scheme 3. 11 compounds from 16-20 through compounds 15 This is about a method of synthesizing. In this embodiment of the present invention, a compound 11Unprotected dioxolanic acid compound obtained through benzyl deprotection (preferably using reductive cleavage, commonly in Pd / C, hydrogen gas, or MeOH at room temperature). 16 It provides in high yield, which is subsequently protected from free hydroxyl groups to isobutyrate groups. This step, for example, compounds 16 This is carried out by reacting isobutyric anhydride with a base (e.g., pyridine and dimethylaminopyridine (DMAP)) in a dichloromethane solvent, initially at 0°C and then reacting at room temperature (e.g., overnight, 12–20 hours, commonly 16 hours), to obtain an isobutyrate-protected compound. 17 It is commonly provided with a yield exceeding 80-85%. compound 17 2-Me-THF is acylated using Mn(II)OAc, diacetoxyiodobenzene, and acetic acid (e.g., initially at 0°C, then overnight or for 12–20 hours at room temperature, often 16 hours, followed by 4 hours at 65°C) to form a compound as a mixture of diastereomers. 18 It was formed. Alternatively, lead tetraacetate Pb(OAc)4 in acetonitrile / base, commonly pyridine or trimethylamine, more commonly pyridine, can be used, and the components are combined at 0°C and then cooled to room temperature for 16 hours (an alternative step is not shown in the reaction scheme below). Subsequently, the compound 18 Among dichloromethane (DCM) tert - Compounds coupled to bromovinyluracil using butyldimethylsilyl trifluoromethanesulfonate (TBDMSOTf), iodotrimethylsilane (TMSI), and 2,4,6-collidine 19 and 20 A mixture of the diastereomers of was provided, and subsequently, said mixture was applied to chromatography (e.g., silica gel using 15-18% EtOAc / hexane) to the compound 20It is commonly provided in a yield exceeding about 45-50%, which is L-BHDU ( 15 It can be easily converted to ). Each of the aforementioned synthesis steps can be performed as a separate step or together in a single reaction vessel.

[0025]

[0026]

[0027]

[0028] In one embodiment, the present invention relates to benzyl-protected dioxolanic acid 9 Benzyl-protected dioxolanic acid 11 It concerns the synthesis of, which is ( S )-Phenylethylamine compound in acetonitrile at room temperature for several hours (e.g., commonly about 4 hours) 9 React with to produce D and L isomer benzyl-protected phenylethylammonium salt 10 A mixture of (a mixture of isomer compounds is also 10M This is accomplished by forming (referred to as), resuspending the solid mixture in ethyl acetate / isopropanyl in a 3:1 ratio (volume) for 4 hours, and filtering. This resuspending process is optionally and preferably repeated several times until the unwanted D isomer is removed from the salt mixture, thereby producing a substantially pure L-isomer compound. 10 (at least 90%, often at least 95%, at least 99%, 99+%, 99.3+% of de Provides ). L-isomer (compound 10 ) is benzyl-protected L-dioxolanic acid at room temperature 11It is exposed to an aqueous acid (e.g., 1N aqueous HCl) for a sufficient amount of time to provide the compound. 11 L-dioxolanic acid deprotected by removing a benzyl protecting group under reducing conditions (e.g., Pd / C, H2). 16 It is easily converted into.

[0029]

[0030]

[0031] In one embodiment, the present invention relates to isobutyrate-protected L-dioxolanic acid 17 isobutyrate-protected nucleoside compounds 20 This relates to a method for synthesizing a compound. 17 A protected L-dioxolane compound acylated by acylating with Mn(II) OAc, diacetoxy iodobenzene, acetic acid in 2-Me-THF (or alternatively lead tetraacetate Pb(OAc)4 in acetonitrile / base, commonly with pyridine or trimethylamine, more commonly with pyridine, combining the components at 0°C and then at room temperature for 16 hours, not shown in the reaction scheme below). 18 It is accomplished by forming a mixture of the diastereomers of. Subsequently, the compound 18 Among dichloromethane (DCM) tert - Coupling with bromovinyl-uracil using butyldimethylsilyl trifluoromethanesulfonate (TBDMSOTf) and then treating with iodotrimethylsilane (TMSI) to form a compound 19 and 20 Provides a mixture of the diastereomers of. compound 19 and 20 A mixture of the diastereomers of is applied to column chromatography to the compound 20 It provides Subsequent compounds 20Silver is exposed to ammonia / methanol (or other suitable base) to remove the isobutyrate protecting group, resulting in the final compound L-BHDU 15 It provides. Each of the aforementioned synthesis steps can be performed as a separate step or together in a single reaction vessel.

[0032]

[0033] In one embodiment, as shown in FIG. 7 and reaction scheme 4, L-BHDU ( 15) is a weak base (e.g., N Compound in THF solvent in the presence of -methylimidazole (NMI) 15 bis(POM)phosphorochloridate ( 21) React with, purify (e.g., chromatography) and crystallize to obtain a compound in a yield of over 40% 22 By providing POM-L-BHDU-MP( 22) It is converted to.

[0034] In embodiments, the present invention also relates to one or more of the compounds comprising the intermediate compounds identified above, or salts thereof, such as pharmaceutically acceptable salts thereof.

[0035] These and / or other embodiments of the present invention can be easily identified by reviewing the presentation and identified discussions and embodiments in the specification below.

[0036] Detailed description of the invention

[0037] The following terms are used to describe the present invention. Where a term remains undefined, it is given the meaning recognized in the art. According to the present invention, conventional chemical synthesis methods and other biological and pharmaceutical techniques within the scope of the art may be used. Such techniques are well known and otherwise sufficiently described in the literature.

[0038] Where a range of values ​​is provided, unless the context clearly indicates otherwise (e.g., in the case of a group including the number of carbon atoms, each number of carbon atoms falling within the range is deemed to be provided), each intervening value up to 1 / 10 of the lower unit between the upper and lower limits of the range, and any other specified value or intervening value within the specified range are understood to be included in the present invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also included in the present invention, provided that the limit values ​​specifically excluded from the specified ranges are included. Where the specified ranges include one or both of the limit values, the ranges excluding either or both of such included limit values ​​are also included in the present invention.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. Any method and material similar or equivalent to that described herein may be used in the practice or testing of the present invention, but preferred methods and materials are described now.

[0040] It should be noted that, as used in this specification and the appended claims, the singular forms “one (a),” “and,” and “the” include plural references unless the context clearly indicates otherwise. When used, the term “about” means an amount within ±5% of the specifically stated amount or number.

[0041] As used herein, the term “compound” refers to any specific chemical compound or intermediate disclosed herein, which is commonly used to produce these nucleoside compounds using the synthesis steps described herein, unless otherwise indicated. β - Refers to L-nucleoside analogs or D and L-dioxolane intermediates, but within the context, may include tautomers, positional isomers, geometric isomers, anomers, and, where applicable, optical isomers (enantiomers) or often diastereomers (two or more chiral centers) of these compounds, as well as their pharmaceutically acceptable salts, solvates, and / or polymorphs. In contextual use, the term compound generally refers to a single compound, but may also include stereoisomers, positional isomers, and / or optical isomers (including racemic mixtures and / or diastereomers) as described herein, as well as other compounds such as specific enantiomers, enantiomerically concentrated compounds, individual diastereomers, or mixtures of these disclosed compounds. Note that if a carbon range is provided for a compound, the range implies that each individual carbon is considered part of the range. For example, C1-C 20 The group describes a group having up to 20 carbons, including a single carbon, two carbon atoms, three carbon atoms, four carbon atoms, etc.

[0042] The terms “salt” or “pharmaceutically acceptable salt” are used throughout this specification to describe salt forms of one or more compounds described herein, where applicable, and are presented to increase the solubility of the compound in the gastric fluid of the patient’s gastrointestinal tract to increase the results of a synthetic reaction, or alternatively, to promote the dissolution and bioavailability of the compound in a particular embodiment in which administration is performed. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids where applicable. Suitable salts include those derived from alkali metals such as potassium and sodium, alkaline earth metals such as calcium and magnesium, and ammonium salts, among numerous other acids well known in the pharmaceutical field. Sodium and potassium salts are particularly preferred as neutralizing salts of the phosphate according to the present invention where applicable.

[0043] The term “pharmaceuticalally acceptable derivative” is used throughout the specification to describe any pharmaceutically acceptable prodrug form (e.g., ester, ether or amide, phosphoramidate, phosphate ester, phosphonate or other prodrug group) that, when administered to a patient, directly or indirectly provides the compound or the active metabolite of the compound.

[0044] As used in the context of the present invention, the term “D-batch” refers to a batch of other intermediates as well as nucleoside compounds according to the present invention that mimic a natural batch of sugar moiety, in contrast to non-naturally occurring nucleosides or “L” batches that describe most compounds used in the chemical synthesis presented herein or used as final products. The term “ β " or " β The term “anomer” is used to describe an intermediate or nucleoside analog according to the present invention in which a nucleoside base is positioned (oriented) on the plane of a dioxolane moiety in the compound.

[0045] The term “diastereomeric excess”, "de" Or “enantiomer excess”, “ ee "Is It is used throughout the specification to describe intermediate compounds or nucleoside compounds having a single diastereomer or a single enantiomer of the compound in an amount of at least about 90%, commonly at least about 95%, preferably at least about 96%, more preferably at least about 97%, even more preferably at least about 98%, and even more preferably at least about 100% or more. Dioxolane nucleoside compounds, such as L-BHDU or derivatives thereof according to the present invention, are generally β It is an L-nucleoside compound analog. Where the compound according to the present invention is referred to herein, unless otherwise specified, the intermediate or final nucleoside compound is presumed to have an L-nucleoside batch and be diastereomericly and / or enantiomerically concentrated (preferably about 100% of L-nucleosides). The terms “diastereomericly pure,” “diastereomericly concentrated,” or “diastereomeric excess” or de " is used to describe a single diastereomer of a compound according to the present invention comprising, on a weight basis, at least 90%, 95%, 96%, 97%, 98%, 99%, 99+%, 99.5%, or 100% excess of the diastereomer, excluding other possible diastereomers.

[0046] The term “stereoselective” is used to describe a synthetic step or series of steps in which a single reactant produces a specific isomer, enantiomer, or diastereomer (of at least two possible isomers or diastereomers) in an amount greater than that of one or more possible isomer(s) or diastereomer(s) from the reactant. In some cases, the stereoselectivity of a reaction can be close to 100%.

[0047] The terms “protector” or “blocker” are used to describe chemical groups or moiety introduced into a molecule by chemical modification of a functional group to obtain chemoselectivity and / or stereoselectivity in a subsequent chemical reaction. Protectors / blockers play a crucial role in providing precursors for the chemical components that provide the compounds according to the present invention. Blockers are commonly used to protect hydroxyl groups on intermediates to form the compounds according to the present invention. Typical blockers are typically used on alcohol or phosphate groups in the present invention and primarily include benzyl groups (removed by hydrolysis), isobutyrate groups (removed by bases), methyl ester groups (removed by bases), acetyl groups (removed in coupling reactions), and pivaloylmethyl ester (POM) groups on phosphate intermediates. Pivaloyloxymethyl ester (POM) groups are also used on the phosphate groups of the final compound POM-L-BHDU-MP to impart prodrug properties to nucleoside compounds to improve bioavailability. These and / or other devices are described in this specification.

[0048] Overview and Results of Chemical Synthesis

[0049] Numerous efforts by various groups to produce chiral pure L-dioxolane often lead to a racemic mixture of D and L-dioxolane, requiring expensive chiral SFC purification to obtain chiral pure D and L-dioxolane. To eliminate chiral SFC separation, Bera S. et al. 19methyl-( R The synthesis of chiral dioxolanes via )-2,2-dimethyl-1,3-dioxolane-4-carboxylate has been reported. However, the reported procedure was ambiguous and required precise analytical techniques to identify D and L stereoisomers, which can sometimes lead to the final product of an undesirable stereotype. In this case, the inventors obtained an L-BHDU standard sample consistent with the L-BHDU synthesized via the protocol reported by Bera S. et al. Subsequently, the inventors found that this procedure, without a standard sample and chiral HPLC, was very difficult to provide the desired isomer of the target compound. To achieve the scalable synthesis of L-BHDU, a new protocol for the synthesis of exclusively chiral pure L-dioxolanes was urgently needed, thus triggering the research leading to the present invention. The present invention focuses primarily on the synthesis of chiral L-dioxolanes with excellent enantiomer excess (ee ≥ 99%). Notably, this procedure may be suitable for the large-scale synthesis of L-derived dioxolane nucleosides such as troxacitabine. 21 It can also facilitate extended structure-activity relationship (SAR) studies of both D and L-dioxane-derived molecules of therapeutic interest.

[0050] Chiral pure L-dioxolane 11 The synthesis of the ) key intermediate is commercially available methyl ( R )-2,2-dimethyl-1,3-dioxolane-4-carboxylate( 7 )cast p - Condensed with 2-benzyloxyacetaldehyde by transketalation in the presence of TSA to form a racemic 1,3-dioxolane derivative, a 1:1 diastereoisomer mixture of dioxolane esters, 8 It was initiated by providing a 72% yield with (Fig. 4, Reaction Scheme 2). Next, a methyl ester ( 8 The hydrolysis of ) is carried out by a 1 M aqueous LiOH solution in THF and acid (9 A mixture of the diastereomers of ) was provided in a 92% yield.

[0051] After that, the goal is Mount Rasemi ( 9 Pure L-dioxolanic acid (with high enantiomer purity (ee > 99%) while avoiding chromatographic separation of ) 11 The goal was to obtain ). However, the selective isolation of L-dioxolane was also difficult. One approach was carboxylic acid 9 The plan was to provide enantiomerically pure L-dioxolane by introducing a chiral auxiliary group through esterification followed by fractional crystallization. However, this strategy unnecessarily increases the additional two-step process of esterification and hydrolysis, which is inefficient for the large-scale synthesis of L-dioxolane. To reduce the additional steps, a racemic method was developed through the formation of a diastereomeric salt with a chiral amine. 9 It was thought to perform the resolution of the partitioning. A notable point is that the compound 9 It is essentially oily, and other fractional crystallization techniques use chiral pure L-dioxolane ( 11 The point is that it may not be suitable for the splitting of ). Singh et al. reported the chiral splitting of racemic amines through chiral pure L-(+)-tartaric acid. 22 Based on the procedure reported by Singh et al., racemic acid ( 9 It was thought that adopting an inverse strategy of chiral partitioning )

[0052] Lacemi 9 To achieve the formation of a diastereomerically pure salt of, several ( R ) or ( S Chiral amines have been used in various solvent systems (below Table 1 (Reference). Racemi Mountain 9 In the reviewed division of, ( S )-with phenylethylamine 9 The formation of diastereoisomer salts of 99.37% Excess of diastereomers ( d pure chiral dioxolane salt having e) 10 generated Table 1 Item 4). Racemic acid 9 Dissolve in acetonitrile and 0.8 equivalents of ( S Treatment with )-phenylethylamine yields a white salt precipitate of the acid amine. The next objective was to determine the chiral purity of the precipitated salt and the stereotype of the diastereoisomer salt. The precipitated diastereoisomer salt was treated with 1 N aqueous HCl to yield free diastereoisomer dioxolane. The chiral purity of the obtained free dioxolane was determined by chiral HPLC, indicating that the optical purity of the precipitated isomer is 85% and the other isomer is present at 15%. Obtained results of diastereoisomer salt formation through Lacemi 9 The splitting of was not encouraging, because the production of optically pure L-BHDU required L-dioxolane with an optical purity of more than 99% (ee ≥ 99%).

[0053] Therefore, racemi 9 The process of diastereomeric salt formation was re-examined. The diastereomeric excess of the precipitated salt (via acetonitrile); de To increase ), the obtained salt was resuspended in EtOAc-IPA (3:1), stirred at room temperature for 4 hours, filtered, and neutralized with a 1 N aqueous HCl solution. The provided dioxolane was re-examined for chiral purity via chiral HPLC, which was greater than 99% with a yield of 38%. ee ( ee It was proven that ≥ 99.3%). Therefore, a dual solvent treatment involving treatment in acetonitrile first followed by treatment with EtOAc:isopropanol (3:1) resulted in an excess of the diastereomer of the precipitated salt ( de improves ) deIt was concluded that ≥ 99.3% was the result, which was unexpected. The subsequent objective was to determine the stereotype of the precipitated salt, namely the D or L-form of dioxolane. X-ray crystals of the precipitated salt were obtained, which showed that the formed diastereoisomer salt was L-dioxolane ( 10 It was confirmed that it matches the stereochemical form of ). The X-ray structure shows that the hydrogen atoms present in carbon-2 (C-2) and C-3 are trans ( Fig. 5b It was revealed that (see reference) this confirms the L-dioxolane salt precipitation.

[0054] however, Table 1 below A single-solvent salt formation approach was attempted with the various chiral amines mentioned, but in each case de The results were not satisfactory. Intermediate 11 To improve the yield, we attempted modified ratios of EtOAc and IPA, but in all efforts, the intermediate 11 A lower yield or lower chiral purity was achieved.

[0055] [Table 1]

[0056] Racemic compounds using various chiral amines 10 result of diastereoisomer salt formation

[0057]

[0058]

[0059] L-dioxolane compound 11 It served as a key intermediate for the synthesis of L-BHDU. 11 If treated with Mn(II)OAc, acetic acid, or diacetoxy iodobenzene in 2-methyl THF, or alternatively with lead(IV) acetate Pb(OAc)4 in the presence of pyridine in acetonitrile, 11 The 2-carboxyl group of the 2-acetoxy intermediate 12 as α / β It is converted to a 74% yield at a ratio of 2:1 (racemite). In addition, 12and the glycosylation coupling of bromovinyl-uracil is Vorbr It was performed under ggen coupling conditions. 23 The bromouracil base treated with trimethylsilyl trifluoromethanesulfonate (TMSOTf) followed by treatment with (iodotrimethylsilane) TMSI is phosphorus Generates silylated bromovinyluracil in situ and acetate intermediate 12 (α / β Combined with (P. 2:1) 13 The coupled β and α Isomers (55% yield) and 14 It provides (18% yield) at a 3:1 ratio. The separation of isomers by column chromatography is desired β- Isomers in 55% yield, α - The isomer was provided in an 18% yield. Finally, the coupled product 13 The deprotection of the 5'-benzyl group was carried out with a 1 M boron trichloride (BCl3) solution in DCM to provide the target compound L-BHDU in 60% yield. This series of reactions is presented in the attached Fig. 4, Reaction Scheme 2.

[0060] Compounds using BCl3 13 It was determined that the exothermic final deprotection of the benzyl group is not suitable for large-scale synthesis. To avoid the use of BCl3, a protecting group substitution strategy was adopted, and the intermediate 11 of 5- O -Benzyl was substituted with an isobutyric acid ester as a hydroxyl protecting group. 11 The debenzylation of was carried out with 5% Pd / C under hydrogenation conditions to produce an intermediate 16 It was provided with a yield of 91%. intermediate 16 by treating with isobutyric anhydride in the presence of pyridine the compound 17 Compound was produced with an 88% yield. 17Following the Asilhwa, the coupling with Bromovinyl-Uracil is the coupled desired (β ) nucleoside 20 (54% yield) and unwanted ( α ) nucleoside 19 (Yield 22%) was provided at a ratio of approximately 2:1 (Fig. 6, Reaction Equation 3 reference).

[0061] 20 Deprotection of the isobutyl ester group was performed in a base (7 N NH3 solution in MeOH) to provide L-BHDU in a 92% yield. 15 The optical rotation and other analysis data were consistent with the previously synthesized standard L-BHDU. 24

[0062] To synthesize the POM-L-BHDU-MP prodrug, first, bis(POM)phosphorochloridate ( 21 Synthesis of ) was performed according to the protocol reported by Hawang Y. et al. 25 Next, L-BHDU( 15 The coupling of ) in THF N - POM chloride in the presence of methylimidazole (NMI) 21) and performed POM-L-BHDU-MP( 22 provided ) with a 47% yield ( Fig. 7, Reaction Equation 4 ).

[0063] Derived conclusion

[0064] To determine the complete biological evaluation of POM-L-BHDU-MP, commercially available methyl ( R )-2,2-dimethyl-1,3-dioxolane-4-carboxylate, 7 An efficient and scalable synthesis method for L-BHDU was developed through this. Chiral ( S Racemosis via )-phenylethylamine 9 Following the selective diastereomeric salt formation of 10 The neutralization of optically pure L-dioxolane, 11Provided (ee ≥ 99%). Compound 11 Silver acetate intermediate 12 It was converted to, and this is Vorbr with bromovinyl-uracil It was used for ggen coupling, followed by benzyl deprotection of the coupled nucleoside at step 7 with a total yield of approximately 4.9% for the target compound. 15 (L-BHDU) was provided. This process eliminates the expensive chiral separation of racemic D and L-dioxolane and is more efficient than the previously reported method for the synthesis of L-BHDU. Bis(POM)phosphorochloridate, 21 Additional coupling with L-BHDU produces an L-BHDU-monophosphate ester prodrug 22 (POM-L-BHDU-MP) was produced. Compared to the previously reported method, the elimination of chiral separation during the synthesis step, the reduction of column chromatography, and the use of economical reagents make the current methodology suitable for the large-scale production of POM-L-BHDU-MP and chiral pure L-dioxolane ( 11 Makes it suitable for the selective synthesis of ).

[0065] The use of Mn(OAc)2 during chemical synthesis of compounds 12 and 18It was unexpectedly discovered to be a particularly effective substitute for Pb(OAc)4 for forming (the conversion of the dioxolanyl moiety of the precursor carboxylic acid to an acetate group at the 4-position) and has become a preferred reagent for carrying out these steps in the scale-up reaction of the present invention. The reactivity of Mn(OAc)2 was found to be at least equivalent to that of Pb(OAc)4 and, in some cases, somewhat greater; despite the overwhelming prevalence of Pb(OAc)4 in conventional chemistry, this reagent possesses other properties that make it substantially more advantageous than Pb(OAc)4 for use in the present invention. Furthermore, Mn(OAc)2 costs about 25% of Pb(OAc)4, providing significant cost savings, particularly in commercial applications. This reagent is more readily available from commercial suppliers due to its much lower reagent toxicity compared to Pb(OAc)4. Compound 12 and 18 In each case, the reaction time for converting the carboxylic acid group to acetate was found to be substantially faster for Mn(OAc)2 (4 hours) compared to Pb(OAc)4 (>14 hours), which will support the rapid synthesis of POM-L-BHDU-MP. Additionally, in commercial applications, handling Mn(OAc)2 is much less difficult than handling Pb(OAc)4, because Pb(OAc)4 is very hygroscopic (moisture-sensitive) and difficult to use in large-scale environments (>25g) and often requires special handling, whereas Mn(OAc)2 is very stable (not moisture-sensitive) and easy to handle.

[0066] Pb(OAc)4 is highly exothermic in reactions on a scale of >50 g, and in the present invention, it was necessary to add the reagent in fractions to control the reaction temperature and carry out the reaction under cooling conditions. This took considerably longer than when using Mn(OAc)2, because the reaction step using Mn(OAc)2 is not exothermic and does not require external cooling. In reactions using Pb(OAc)4, pyridine is required in equimolar amounts, and removing pyridine from the final work product after the reaction is complete is difficult and produces a particularly pungent odor in large-scale reactions. Since the Mn(OAc)2 reaction does not require pyridine (or any amine), it substantially reduces the burden of compound purification and final processing. Additional considerations include the fact that the product obtained from the Pb(OAc)4 reaction must be purified by column chromatography to be sufficiently pure for use in the next step, whereas Mn(OAc)2 results in a nearly clean conversion and the crude product can be readily used in the next step reaction, thus avoiding column chromatography purification. This is particularly important in the commercial application of the present invention.

[0067] In the present invention, under large-scale Pb(OAc)4 reaction conditions, colloidal solutions within the reaction mixture and during the work-up process cause difficulties in processing the reaction mixture and disrupting the colloidal state of the reaction mixture. The formation of such colloidal solutions can often impede the reaction yield. While Pb(OAc)4 reaction conditions are not environmentally friendly, conditions utilizing Mn(OAc)2 are environmentally friendly and substantially safe for the environment. Therefore, Mn(OAc)2 is a substantially superior reagent for scalable synthesis to be used in the present invention.

[0068] Examples

[0069] Experiment Section

[0070] General analysis methods

[0071] Reagents and anhydrous solvents were purchased from commercial sources and used without further purification. Moisture-sensitive reactions were performed using oven-dried glassware under a nitrogen or argon atmosphere. Reactions were visualized using a Spectroline UV lamp (254 nm) and monitored on thin-layer chromatography plates (TLC silica gel GF 250 micron) colored with a 15% sulfuric acid solution in methanol. Column chromatography was performed on silica gel 60 Å, 40–63 μM (230 x 400 mesh, Sorbent Technologies). Preparative normal-phase chromatography was performed on a CombiFlash Rf 150 (Teledyne Isco) equipped with a pre-filled RediSep Rf silica gel cartridge or on a RediSep®gold C18 reverse-phase column. Melting points were recorded on a Mel-temp II laboratory instrument and were uncorrected. Nuclear magnetic resonance spectra were prepared using tetramethylsilane as an internal standard. 1 For H NMR, 500 MHz, 31 For P NMR, 202 MHz, 131C NMR was recorded at 125 MHz using a Varian Inova 500 spectrometer. Chemical shifts (δ) are denoted as s (singlet), bs (broad singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd (double-doublet), and dt (double-triplet). Optical rotation was measured using a JASCO DIP-370 digital polarimeter. High-resolution mass spectrometry (HRMS) spectra were measured using a Bruker Ultra-high resolution QTOF MS Impact II spectrometer. Samples were injected at a rate of 3 μL / min, and spectra were obtained in cationized or anionized mode at a typical resolution of over 20,000. The purity of all tested compounds is ≥95%, as determined by elemental analysis or HPLC / UV. Elemental analysis was performed by Atlantic Microlab Inc., Norcross, GA. HPLC / UV was measured using a Waters HPLC coupled to a photodiode array. 0.5 mg / mL sample 5 in methanol, acetonitrile, or a DMSO / MeOH (0.5:10) mixture μ L was injected, and XBrigde C18, 3.5 at 25℃ at a flow rate of 0.8 mL / min. μ Use a m, (4.6 X 150) mm column, or UPLC BEH C18, 1.7 at 50℃ with a flow rate of 0.55 mL / min μA m (100 X 2.1) mm column was used. The mobile phase consisted of a mixture of A, 10 mM ammonium acetate in water, and B, acetonitrile (ACN), and A, 0.05% formic acid (FA) in water, and B, 0.05% acetonitrile (ACN). Purity was expressed as absorbance (%) on the Max plot. The optical purity of the chiral intermediate and final compounds was determined by chiral HPLC. Chiral HPLC / UV was measured using a Waters HPLC coupled to a photodiode array. 10 μL of a 0.5 mg / mL sample in methanol was injected, and a CHIRALCEL OX-H, 5 μmm (4.6 X 250 mm) column was used at 30°C with a flow rate of 3.0 mL / min.

[0072] Methyl (4 R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylate (8): 2-(benzyloxy)acetaldehyde (100 g, 665.91 mmol) and methyl ( R )-2,2-dimethyl-1,3-dioxolane-4-carboxylate( 7 In a solution of , 106.65 g, 665.91 mmol) p -Toluenesulfonic acid or Dowex 50W X8 was added (5.06 g, 26.63 mmol), and the mixture was stirred at 80°C for 2 hours. (The reaction mixture turned dark black.) The reaction mixture was cooled to room temperature and volatile substances were evaporated, and the crude mixture was purified using flash silica gel column chromatography (12-18% EtOAc / hexane) as the eluent to obtain methyl (4 R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylate 8 An inseparable mixture of was obtained as a pale yellow oil. Yield (122.5 g, 72%); 1 H NMR (500 MHz, CDCl3) δ7.37-7.32 (m, 11H), 7.30-7.27 (m, 2H), 5.33 (t, J = 4.0 Hz, 1H), 5.22 (t, J= 4.5 Hz, 1H), 4.68-4.58 (m, 7H), 4.31 (dd, J = 8.2, 7.3 Hz, 1H), 4.24 (dd, J = 8.6 & 3.9 Hz, 1H), 4.13-4.09 (m, 1H), 3.99 (dd, J = 8.3 & 5.4 Hz, 1H), 3.78 (s, 3H), 3.75 (s, 3H), 3.71 (d, J = 3.8 Hz, 1H), 3.67-3.61 (m, 2H), 3.59 (d, J = 4.1 Hz, 1H); 13 C NMR (125 MHz, CDCl3) δ171.35, 171.1, 128.49, 128.46, 127.91, 127.83, 127.81, 127.79, 104.87, 104.33, 74.10, 73.90, 73.80, 73.74, 70.87, 70.30, 68.71, 68.31, 52.49; HRMS-ESI (m / z): [M+Na] + C 13 H 16 Calculated value for O5Na 275.0895; measured value 275.0883.

[0073] (4 R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylic acid (9): Methyl (4) in THF (250 mL) at 0℃ R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylate 8 1 M aqueous LiOH (198 mL) was added dropwise to a solution of (60 g, 237.84 mmol), the mixture was slowly raised to room temperature, and stirred for 36 hours. THF was removed under reduced pressure, and the mixture was washed with EtOAc (3 x 75 mL). The separated aqueous layer was cooled to 0°C, acidified to pH ~3 with 28% aqueous HCl, and extracted with DCM (3 x 125 mL). The combined organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated to obtain racemic acid (4 R)-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylic acid 9 A light brown viscous liquid was obtained. Yield (52.5 g, 92%); 1 H NMR (500 MHz, CDCl3) δ7.40-7.32 (m, 10H), 7.31-7.27 (m, 1H), 5.34 (t, J = 3.6 Hz, 1H), 5.20 (s, 1H), 4.74 (d, J = 12.1 Hz, 1H), 4.70-4.64 (m, 3H), 4.61 (s, 2H), 4.42-4.34 (m, 2H), 4.15-4.09 (m, 2H), 4.04 (dd, J = 8.4 & 5.4 Hz, 1H), 3.80 (dd, J = 11.4 & 1.7 Hz, 1H), 3.76 (d, J = 11.4 Hz, 1H), 3.61 (dd, J = 5.9 & 3.6 Hz, 2H); 13 C NMR (125 MHz, CDCl3) δ174.76, 173.56, 137.68, 136.02, 128.77, 128.53, 128.28, 127.91, 127.85, 104.44, 104.25, 74.28, 73.83, 73.65, 70.48, 70.19, 68.27, 67.50; HRMS-ESI (m / z): [M+Na] + C 12 H 14 Calculated value for O5Na 261.0739; measured value 261.0739.

[0074] (2 S ,4 R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylate-( S )-1-phenylethane-1-aluminum (10): Methyl (4 in ACN (50 mL) R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylate 9 A solution of (25 g, 105.0 mmol) was heated to 70°C for 20 minutes. After that, (S )-(-)-1-phenylethylamine (10.80 g, 89.25 mmol) was added dropwise. The mixture was heated at the same temperature of 70°C for 2 hours, then slowly cooled to room temperature and stirred for 3 hours to yield a white solid precipitate. The precipitate was filtered. To remove unwanted D-dioxolane salt (15%) from the precipitate, the filtered solid was resuspended in a mixture of EtOAc:isopropanol (50 mL, 3:1), stirred at room temperature for 4 hours, and then filtered. 10 The purity of the diastereomers was monitored by TLC and NMR, and the process was repeated three times until the unwanted isomer (D-dioxolane salt) was completely removed from the solid precipitate. Compound 10 It was obtained as a white amorphous solid. Yield (11.5 g, 32%); 1 H NMR (500 MHz, CDCl3) δ8.37 (bs, 3H), 7.35-7.25 (m, 10H), 5.03 (t, J = 3.9 Hz, 1H), 4.57-4.50 (m, 2H), 4.27-4.21 (m, 2H), 4.04 (t, J = 7.7 Hz, 1H), 3.63 (t, J = 7.1 Hz, 1H), 3.46-3.43 (m, 2H), 1.49 (d, J = 6.8 Hz, 3H); 13 C NMR (125 MHz, CDCl3) δ176.74, 139.31, 137.81, 128.91, 128.50, 128.47, 127.89, 127.86, 126.63, 103.01, 75.66, 73.373, 70.84, 68.69, 50.93, 21.13.

[0075] (2 S ,4 R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylic acid (11): (2 S ,4 R)-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylate-( S )-1-phenylethane-1-amium, 10 Dissolve (11.2 g, 31.18 mmol) in DCM (200 mL), and cold 1 The organic layer was washed sequentially with aqueous HCl (20 mL), water (100 mL), and finally brine (100 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (2S,4R)-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylic acid 11 was provided as a colorless viscous liquid. Yield (7.1 g, 95%); [α] 25 D = +20.82 (c 0.5, CHCl3) 1 H NMR (500 MHz, CDCl3) δ7.61 (bs, 1H), 7.36-7.31 (m, 4H), 7.30-7.25 (m, 1H), 5.32 (t, J = 3.5 Hz, 1H), 4.69-4.66 (m, 1H), 4.60 (s, 2H), 4.34 (t, J = 7.9 Hz, 1H), 4.02 (dd, J = 8.4 & 5.4 Hz, 1H), 3.60 (dd, J = 5.9 & 3.6 Hz, 2H); 13 C NMR (125 MHz, CDCl3) δ176.74, 139.31, 137.81, 128.91, 128.50, 128.47, 127.89, 127.86, 126.63, 103.01, 75.66, 73.373, 70.84, 68.69, 50.93, 21.13; HRMS-ESI (m / z): [M+H] + C 12 H 14 Calculated value for O5Na 261.0739; measured value 261.0729.

[0076] (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate (12): In 2-Me-THF (12 mL) (2S ,4 R )-2-((benzyloxy)methyl)-1,3-dioxolane-4-carboxylic acid( 11 A stirred solution of (0.5 g, 2.09 mmol) was degassed with argon, then acetic acid (0.25 mL, 42.18 mmol), Mn(II)OAc (994 mg, 5.74 mmol), and diacetoxy iodobenzene (2 g, 6.27 mmol) were added, and degassed again for 3 minutes. The reaction mixture under argon was heated to 65°C for 4.5 hours. The reaction mixture was cooled to room temperature and filtered through a Celite bed. The resulting filtrate was diluted with EtOAc (60 mL); and washed sequentially with saturated Na2S2O3 (15 mL), cold 2M NaHCO3 (15 mL) water (25 mL), and finally saturated brine (30 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography using 16-19% EtOAc / hexane as the eluent (2 S A mixture of the diastereomers of )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate is a colorless liquid ( 12 ) obtained. Yield (385 mg, 72%); 1 H NMR (500 MHz, CDCl3) δ7.37-7.27 (m, 8H), 6.39 (dd, J = 4.4 & 2.1 Hz, 1H), 6.33 (d, J = 3.7 Hz, 0.6H), 5.38-5.36 (m, 1H), 5.30-5.27 (m, 0.7H), 4.64-4.59 (m, 3H), 4.23 (dd, J = 9.5 & 4.7 Hz, 1H), 4.18 (d, J = 9.6 Hz, 0.6H), 3.99-3.93 (m, 2H), 3.67-3.57 (m, 4H), 2.09 (s, 3H), 2.03 (s, 2H); 13 C{ 1H} NMR (126 MHz, CDCl3) δ170.4, 137.8, 128.5, 127.8, 105.3, 103.8, 94.6, 94.1, 73.8, 73.8, 73.7, 71.4, 71.1, 70.9, 70.2, 21.2; HRMS-ESI ( m / z ): [M + Na] + Calculated value [C 13 H 16 O5] + 275.0895; Measured value 275.0885.

[0077] (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate (12) (alternative synthesis): In acetonitrile (15 mL) (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate , 11 Pb(OAc)4 (4.46 g, 1.48 mmol) was added in fractions at 0°C to a stirred solution of (2.0 g, 8.39 mmol) and anhydrous pyridine (0.95 mL, 12.08 mmol), and the mixture was stirred continuously at room temperature for 16 hours. The reaction mixture was filtered through a Celite bed. The resulting filtrate was diluted with water (100 mL) and extracted with EtOAc (2 x 50 mL). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography using 15-18% EtOAc / hexane as the eluent (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate 12 A colorless liquid was obtained. Yield (1.35 g, 74%); 1 H NMR (500 MHz, CDCl3) δ7.37-7.27 (m, 8H), 6.39 (dd, J = 4.4 & 2.1 Hz, 1H), 6.33 (d, J= 3.7 Hz, 0.6H), 5.38-5.36 (m, 1H), 5.30-5.27 (m, 0.7H), 4.64-4.59 (m, 3H), 4.23 (dd, J = 9.5 & 4.7 Hz, 1H), 4.18 (d, J = 9.6 Hz, 0.6H), 3.99-3.93 (m, 2H), 3.67-3.57 (m, 4H), 2.09 (s, 3H), 2.03 (s, 1.6H); 13 C NMR (125 MHz, CDCl3) δ170.04, 137.75, 128.51, 127.84, 105.29, 103.81, 94.58, 94.10, 73.82, 73.82, 73.71, 71.38, 71.10, 70.87, 70.16; HRMS-ESI (m / z): [M+H] + C 13 H 16 Calculated value for O5Na 275.0895; measured value 275.0884.

[0078] 1-((2 S ,4 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl)-5-(( E )-2-bromovinyl)pyrimidine-2,4(1H,3H)-dione (13): In dry DCM (15 mL) ( E TBDMSOTf (1.37 mL, 5.99 mmol) and 2,4,6-collidin (0.97 mL, 5.99 mmol) were added to a suspension of )-5-bromovinyluracil (500 mg, 2.30 mmol) at room temperature and stirred for 30 minutes. To the resulting solution, in DCM (20 mL) 12 After slowly adding a solution of (580 mg, 2.30 mmol), TMSI (0.36 mL, 2.53 mmol) was added. The mixture was stirred at room temperature for 3 hours and quenched with a saturated aqueous Na2S2O3 solution. The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (1:4 EtOAc / hexane as eluent) to remove the unwantedα -Isomers 14 (185 mg, 18%) as a white solid, desired β -Isomers 13 (525 mg, 55%) was provided as a white solid. 1 H NMR (500 MHz, CDCl3) δ8.27 (brs, 1H), 7.91 (s,1H), 7.40-7.29 (m, 6H), 6.37-6.31 (m, 2H), 5.12 (s, 1H), 4.69-4.61 (m, 2H), 4.23 (d, J = 10.1 Hz, 1H), 4.15 (dd, J = 10.2 & 5.3 Hz, 1H), 3.84 (dd, J = 5.6 & 1.8 Hz, 2H); 13 C NMR (125 MHz, CDCl3) δ160.83, 149.26, 138.0, 136.93, 128.82, 128.52, 128.13, 127.94, 111.62, 110.01, 104.91, 81.44, 74.36, 71.99, 67.99; HRMS-ESI (m / z): [M+H] + C 17 H 17 Calculated value for BrN2O5Na 431.0219; measured value 431.0209.

[0079] 5-(( E )-2-Bromovinyl)-1-((2 S ,4 S )-2-(hydroxymethyl)-1,3-dioxolane-4-yl)pyrimidine-2,4(1 H ,3 H )-Dion (15): 1-((2 in DCM (2 mL) at -78℃ S ,4 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl)-5-(( E )-2-bromovinyl)pyrimidine-2,4(1 H ,3 H )-Dion, 131 M BCl3 (1.34 mL, 1.34 mmol) in DCM was added dropwise to a solution of (500 mg, 1.22 mmol) and stirred for 30 minutes at the same temperature. Afterward, the reaction mixture was quenched with MeOH (1 mL), heated to room temperature, and the inorganic salt was filtered. The organic layer was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM / MeOH, 9:1) to obtain 1-((2 S ,4 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl)-5-(( E )-2-bromovinyl)pyrimidine-2,4(1 H ,3 H )-Dion, 15 (235 mg, 60%) was provided as a white solid. [α] 23 D = -6.2 (c 0.2.7, MeOH); 1 H NMR (500 MHz, CDCl3) δ11.61 (brs, 1H), 8.16 (s, 1H), 7.22 (d, J = 13.6 Hz, 1H), 6.82 (d, J = 13.6 Hz, 1H), 6.21 (d, J = 5 Hz, 1H), 5.31 (t, J = 6.0 Hz, 1H), 4.97-4.94 (m, 1H), 4.31 (d, J = 9.9 Hz, 1H), 4.09 (dd, J = 9.9, 5.6 Hz, 1H), 3.74-3.67 (m, 2H); 13 C NMR (125 MHz, CDCl3) δ162.25, 150.10, 140.10, 130.35, 110.18, 107.01, 105.83, 81.15, 71.07, 60.36; HRMS-ESI (m / z): [M+H] + C 10 H 11 Calculated value for BrN2O5Na 340.9749; measured value 340.9738.

[0080] (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate (16): In MeOH (20 mL) at ambient temperature (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate( 11 A suspension of , 3.0 g, 12.60 mmol) and 5% Pd / C (150 mg) was hydrogenated in a Parr hydrogenator at 8 psi for 3 hours. The mixture was diluted with methanol (20 mL) and passed through a Celite bed. The filtrate was concentrated under reduced pressure (2 S ,4 R )-2-(hydroxymethyl)-1,3-dioxolane-4-carboxylic acid 16 It was obtained as a colorless viscous liquid. Yield (1.72 g, 92%); 1 H NMR (500 MHz, CDCl3) δ5.26 (dt, J = 2.9, 11.0 Hz, 1H), 4.71 (ddd, J = 5.8, 7.3 & 17.4 Hz, 1H), 4.43-4.32 (m, 1H), 4.04 (ddd, J = 5.7, 8.4 & 29.7 Hz, 1H), 3.78-3.74 (m, 2H); HRMS-ESI (m / z): [M+H] + Calculated value for C5H8O5 148.0372; measured value 148.0364.

[0081] (2 S ,4 R )-2-((isobutyryloxy)methyl)-1,3-dioxolane-4-carboxylic acid (17): In dry pyridine (15 mL) at 0℃ (2 S ,4 R )-2-(hydroxymethyl)-1,3-dioxolane-4-carboxylic acid 16 To a solution of (2.0 g, 13.50 mmol), isobutyric anhydride (3.2 mL, 20.26 mmol) was added, then DMAP (8 mg) was added, and the mixture was heated at room temperature. The mixture was stirred for 16 hours. The reaction mixture was diluted with DCM (60 mL), washed with water (2 x 25 mL), and finally with brine (25 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography using 60-65% EtOAc / hexane as the eluent (2 S ,4 R )-2-[(isobutyryloxy)methyl)]-1,3-dioxolane-4-carboxylic acid 17 was provided as a colorless liquid. Yield (2.60 g, 88%); 1 H NMR (500 MHz, CDCl3) δ5.38 (t, J = 3.7 Hz, 1H), 4.67 (dd, J = 5.1, 7.1 Hz, 1H), 4.30-4.25 (m, 1H), 4.18 (t, J = 3.6 Hz, 2H), 4.02 (dd, J = 5.1 & 8.4 Hz, 1H), 2.60 (p, J = 7.0 Hz, 1H), 1.18 (d, J = 7.0 Hz, 6H); 13 C NMR (125 MHz, CDCl3) δ173.04, 135.94, 128.88, 128.61, 128.30, 104.21, 74.34, 70.60, 67.30; HRMS-ESI (m / z): [M+H] + C9H 14 Calculated value for O6Na 241.0688; measured value 241.0673.

[0082] (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate (18):A stirred solution of (2S,4R)-2-((isobutyryloxy)methyl)-1,3-dioxolane-4-carboxylic acid (5.0 g, 2.09 mmol) in 2-Me-THF (150 mL) was degassed with argon, after which acetic acid (2.7 mL, 4.58 mmol), Mn(II)OAc (10.7 g, 6.19 mmol), and diacetoxy iodobenzene (22.15 g, 6.88 mmol) were added and degassed again for 3 minutes. The reaction mixture under argon was heated to 65°C for 4.5 hours. The reaction mixture was cooled to room temperature and filtered through a Celite bed. The obtained filtrate was diluted with EtOAc (160 mL); The product was washed sequentially with saturated Na2S2O3 (150 mL), cold 2M NaHCO3 (150 mL) water (200 mL), and finally saturated brine (150 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography using 16-19% EtOAc / hexane as the eluent to obtain (2S)-4-acetoxy-1,3-dioxolane-2-yl)methyl isobutyrate ( 18 A mixture of the diastereomers of ) was obtained. Yield (4.5 mg, 84%); 1 ¹H NMR (500 MHz, CDCl₃) (mixture of diastereomers) δ 6.34–6.28 (m, 1H), 5.35, 5.25 (t, J = 4.0 Hz, 1H), 4.33-3.90 (m, 4H), 2.58-2.51 (m, 1H), 2.04-2.02 (m, 3H), 1.13-1.11 (m, 6H); 13 C NMR (125 MHz, CDCl3) δ176.5, 176.5, 170.2, 170.1, 103.6, 102.6, 102.3, 94.5, 94.0, 71.3, 70.8, 64.1, 63.5, 63.4, 63.2, 33.8, 21.1, 21.0, 18.9, 18.8; HRMS (ESI-TOF) m / z: [M + Na] + C 10 H 16Calculated value for O6Na 255.0839; measured value 255.0834.

[0083] (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate (18) (alternative synthesis): Among acetonitrile (2 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl acetate 17 Pb(OAc)4 (3.80 g, 8.58 mmol) was added in fractions at 0°C to a solution of (1.5 g, 6.87 mmol) and dry pyridine (0.78 mL, 9.90 mmol), and stirred at room temperature for 16 hours. Afterward, the reaction mixture was passed through a Celite bed. The filtrate was diluted with EtOAc (100 mL), washed with water (50 mL), and finally with brine (50 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography (12-16% EtOAc / hexane) (2 S )-4-acetoxy-1,3-dioxolane-2-yl)methylisobutyrate 18 A colorless liquid was obtained. Yield (1.23 g, 78%); 1 ¹H NMR (500 MHz, CDCl₃) (mixture of diastereomers) δ 6.34–6.28 (m, 1H), 5.35, 5.25 (t, J = 4.0 Hz, 1H), 4.33-3.90 (m, 4H), 2.58-2.51 (m, 1H), 2.04-2.02 (m, 3H), 1.13-1.11 (m, 6H); 13 C NMR (125 MHz, CDCl3) δ176.5, 176.5, 170.2, 170.1, 103.6, 102.6, 102.3, 94.5, 94.0, 71.3, 70.8, 64.1, 63.5, 63.4, 63.2, 33.8, 21.1, 21.0, 18.9, 18.8; HRMS (ESI-TOF) m / z: [M + Na] + C 10 H 16Calculated value for O6Na 255.0839; measured value 255.0834.

[0084] 1-((2 S ,4 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl)-5-(( E )-2-bromovinyl)pyrimidine-2,4(1 H ,3 H )-Dion (20): In dry DCM (12 mL) ( E TBDMSOTf (0.96 mL, 4.21 mmol) and 2,4,6-collidin (0.55 mL, 4.21 mmol) were added to a suspension of )-5-bromovinyluracil (350 mg, 1.62 mmol) at room temperature. The reaction mixture was stirred for 30 minutes. Afterwards, in DCM (10 mL) to the resulting solution 18 After adding a solution of (375 mg, 1.78 mmol), TMSI (0.25 mL, 1.78 mmol) was added. The mixture was stirred at room temperature for 3 hours and quenched with a saturated aqueous Na2S2O3 solution. The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (15-18% EtOAc / hexane) to remove unwanted α -Isomers 19 (140 mg, 22%) as a white solid, desired β -Isomers 20 (340 mg, 54%) was provided as a white solid. 1 H NMR (500 MHz, CDCl3) δ8.27 (brs, 1H), 7.91 (s,1H), 7.40-7.29 (m, 6H), 6.37-6.31 (m, 2H), 5.12 (s, 1H), 4.69-4.61 (m, 2H), 4.23 (d, J = 10.1 Hz, 1H), 4.15 (dd, J = 10.2 & 5.3 Hz, 1H), 3.84 (dd, J = 5.6 & 1.8 Hz, 2H);13 C NMR (125 MHz, CDCl3) δ160.83, 149.26, 138.0, 136.93, 128.82, 128.52, 128.13, 127.94, 111.62, 110.01, 104.91, 81.44, 74.36, 71.99, 67.99; HRMS-ESI (m / z): [M+H] + C 17 H 17 Calculated value for BrN2O5Na 431.0219; measured value 431.0210.

[0085] 5-(( E )-2-Bromovinyl)-1-((2 S ,4 S )-2-(hydroxymethyl)-1,3-dioxolane-4-yl)pyrimidine-2,4(1 H ,3 H )-Dion (15): 1-((2 in 5 mL of MeOH in a sealed tube at 0 ℃ S ,4 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl)-5-(( E )-2-bromovinyl)pyrimidine-2,4(1 H ,3 H To a stirred solution of )-dione (250 mg, 0.64 mmol), a 7 N NH3 solution in MeOH was added, and the mixture was stirred at room temperature for 16 hours. Volatile substances were removed under reduced pressure, and the resulting residue was triturated with diethyl ether (2 x 5 mL) to 1-((2 S ,4 S )-2-((benzyloxy)methyl)-1,3-dioxolane-4-yl)-5-(( E )-2-bromovinyl)pyrimidine-2,4(1 H ,3 H )-Dion( L-BHDU, 15 )(180 mg, 88%) was provided as a milky white solid. [α] 23 D = -6.1 (c 0.2.7, MeOH); 1H NMR (500 MHz, CDCl3) δ11.61 (brs, 1H), 8.16 (s, 1H), 7.22 (d, J = 13.6 Hz, 1H), 6.82 (d, J = 13.6 Hz, 1H), 6.21 (d, J = 5 Hz, 1H), 5.31 (t, J = 6.0 Hz, 1H), 4.97-4.94 (m, 1H), 4.31 (d, J = 9.9 Hz, 1H), 4.09 (dd, J = 9.9, 5.6 Hz, 1H), 3.74-3.67 (m, 2H); 13 C NMR (125 MHz, CDCl3) δ162.25, 150.10, 140.10, 130.35, 110.18, 107.01, 105.83, 81.15, 71.07, 60.36; HRMS-ESI (m / z): [M+H] + C 10 H 11 Calculated value for BrN2O5Na 340.9749; measured value 340.9741.

[0086] L-BHDU-MP's non-POM prodrug (POM-L-BHDU-MP, 22): L-BHDU in dry THF (3 mL) 15 , 80 mg, 0.25 mmol) and N - In a stirred solution of methylimidazole (0.16 mL, 2.0 mmol), bis(POM)phosphorochloridate at 0°C 25 21 (500 mg, 1.23 mmol) was dissolved in dry THF (3 mL) and added, and stirred for 15 minutes. Afterward, the reaction was heated to room temperature and stirred for 3 hours. The mixture was quenched with methanol, and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography (0.5% MeOH / DCM). 22 It was provided as a colorless viscous oil, which was crystallized in DCM-pentane to provide a milky white solid. Yield: (75 mg, 47%). Melting point: 80-85℃; 1H NMR (500 MHz, CDCl3) δ8.97 (bs, 1H), 7.71 (s, 1H), 7.45 (d, J = 13.6 Hz, 1H), 6.79 (d, J = 13.6 Hz, 1H), 6.35 (d, J = 4.5 Hz, 1H), 5.72-5.65 (m, 4H), 5.15 (s, 1H), 4.43-4.39 (m, 1H), 4.35-4.31 (m, 1H), 4.26-4.21 (m, 1H), 4.20-4.17 (m, 1H), 1.23 (s, 18H); 31 P NMR (202 MHz, CDCl3) δ-3.02; 13 13C NMR (125 MHz, CDCl3) δ176.9, 161.1, 149.5, 137.3, 128.4, 112.2, 110.6, 102.9, 83.2, 81.4, 71.6, 65.1, 38.9, 26.9; Analysis. C 22 H 32 BrN2O 12 Calculated values ​​for P: C, 42.12; H, 5.14; N, 4.47. Measured values: C, 42.34, H, 5.23; N, 4.26; HRMS (EI) (C 22 H 32 BrN2O 12 P+H) + Calculated value for 627.0954, measured value m / z 627.0953.

[0087] Supplementary Information

[0088] Supplementary information can be found for free on the website pubs.acs.org.

[0089] All intermediates and final compounds 3 - 28 X-ray crystallography data and 1 H, 13 C, and 19 Includes a copy of the F NMR spectrum (PDF).

[0090] Access code

[0091] CCDC includes supplementary crystallographic data for this paper. These data can be obtained from the website ccdc.cam.ac.uk / data_request / cif, by emailing data_request@ccdc.cam.ac.uk, or by contacting The Cambridge Crystallographic Data Centre, 12 Union Road, Cambridge CB2 1EZ, UK; fax: +44 1223 336033.

[0092] References

[0093]

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[0095]

Claims

Claim 1 chemical structure 11 Compounds according to compound 9 ( α and β (mixture of isomers) As a method of synthesizing from, compound 9 in acetonitrile as a solvent ( S )- Complexed with phenylethylamine to form phenylethylammonium salt, and D and L isomers 10M The method comprises the step of precipitating and / or crystallizing a phenylethylammonium salt (acid amine salt) as a mixture of, and the D and L isomers 10M A compound having at least 95% diastereomeric excess obtained by dissolving a salt mixture of in 3:1 ethyl acetate:isopropanol and then precipitating it at least two more times. 10 It provides, and compounds 10 Compound having at least 95% diastereomer excess after washing with aqueous acid 11 A method that provides. Claim 2 In claim 1, the acid amine salt 10M The above mixture of D and L isomers is dissolved in 3:1 ethyl acetate:isopropanol and precipitated at least three times, and the resulting compound 10 A method having at least 95% excess of diastereomers. Claim 3 In paragraph 2, the generated compound 10 A method having at least 99% excess of diastereomers. Claim 4 Method according to claim 1, wherein the aqueous acid is 1.0 N HCl. Claim 5 In any one of claims 1 to 4, the compound 11 A compound is formed by reacting Mn(OAc)2, diacetoxy iodobenzene, and a weak acid in a solvent. 12 A method of forming. Claim 6 In any one of claims 1 to 4, the compound 11 A compound formed by reacting lead tetraacetate (Pb(OAc)4) and a weak base 12 A method of forming. Claim 7 In paragraph 5 or 6, a compound using tert-butyldimethylsilyl trifluoromethanesulfonate (TBDMSOTf) 12 Coupling to bromovinyl uracil and then treating with iodotrimethylsilane (TMSI) to form a compound 13 and compounds 14 A method of providing a mixture of. Claim 8 In claim 7, the above compound 13 and 14 Separating the mixture using column chromatography to form compounds 13 A method that provides. Claim 9 In paragraph 8, the above compound 13 Compound LBHDU ( 15 A method that provides ). Claim 10 In paragraph 9, the above LBHDU ( 15 ) is reacted with bis(POM)phosphorochloridate in a weak base solvent to POM- L -BHDU-MP( 22 A method that provides ). Claim 11 chemical formula 20 Compounds according to compound 16 As a method of synthesizing from, a compound 16 The compound protects the free hydroxyl group with an isobutyrate group. 17 Step of providing; compound 17 A compound formed by reacting Mn(OAc)2 with diacetoxy iodobenzene in a weak acid solvent. 18 Provide, or react compound 17 with lead tetraacetate (Pb(OAc)4) and a weak base to form a compound 18 A step of providing compound 18 by coupling it to bromovinyl uracil using tert-butyldimethylsilyl trifluoromethanesulfonate (TBDMSOTf) and then treating it with iodotrimethylsilane (TMSI) to form the compound 19 and compounds 20 A step of providing a mixture of compounds; separating the mixture of compounds 19 and 20 using chromatography to form compounds 20 A method comprising the step of providing Claim 12 In paragraph 11, the compound 17 A compound formed by reacting Mn(OAc)2 with diacetoxy iodobenzene in a weak acid solvent. 18 A method that provides. Claim 13 A method of separating compounds 19 and 20 using silica gel column chromatography in claim 11. Claim 14 In paragraph 11, the compound 20 By deprotecting and removing the isobutyrate protector, L-BHDU ( 15 A method that provides ). Claim 15 In claim 14, the above isobutyrate group is removed in ammonia / methanol. Claim 16 In paragraph 11, the compound 16 silver compounds 11 From, compound 11 is reacted with hydrogen gas using Pd / C in methane as a catalyst to form compound 11 A method manufactured by removing the benzyl protecting group. Claim 17 In paragraph 14 or 15, L-BHDU ( 15) A method of providing POM-L-BHDU-MP by reacting with bis(POM)phosphorochloridate in the presence of a weak base in a solvent. Claim 18 In paragraph 17, the method wherein the weak base is N-methylimidazole (NMI) and the solvent is tetrahydrofuran (THF). Claim 19 compound 9 compound 7 A method of manufacturing from, a compound 7 React with 2-(benzyloxy)acetaldehyde in the presence of acid in a solvent to form a compound 8 Step of forming; and compound 8 Compound as a racemic mixture by hydrolyzing the methyl ester group of 9 A method comprising the step of providing Claim 20 In paragraph 19, the above compound 8 A method for purifying using chromatography. Claim 21 A method according to any one of claims 1 to 20, wherein the reaction steps are performed separately or in a single pot. Claim 22 Compound 8, Compound 9 , compound 10M , compound 10 , compound 11 , compound 12, compound 13 A compound selected from the group consisting of mixtures thereof. Claim 23 compound 16 , compound 17 , compound 18 , compound 20 A compound selected from the group consisting of mixtures thereof.

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