JP2009298785A - Synthesis of acyclic nucleoside derivative - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide acyclic nucleoside derivatives, a method for production thereof, and the new intermediates thereof. <P>SOLUTION: There are provided acyclic nucleoside derivatives using the new intermediates represented by formulas A and B, and also a method for production thereof. In formulas A and B, R<SB>6</SB>and R<SB>7</SB>are each 1-7C alkyl, benzyl or the like; R<SB>8</SB>is 1-21C saturated or mono-unsaturated optionally-substituted alkyl; R<SB>12</SB>is -CH(Ph)<SB>2</SB>, -C(Ph)<SB>3</SB>or the like; and X<SB>2</SB>is a leaving group or OH. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to the field of antiviral agents, and in particular to derivatives of acyclic nucleosides useful for herpes and retroviral infections, processes for their preparation and novel intermediates thereof.

  The practical utility of many acyclic nucleosides is limited due to their relatively mild pharmacokinetics. Many prodrug approaches are being explored in an effort to improve the general bioavailability of acyclic nucleosides. One of these approaches involves preparing ester derivatives of one or more hydroxy groups on acyclic side chains, especially aliphatic esters.

  Patent Document 1 describes 9- [4-hydroxy- (2-hydroxymethyl) butyl] guanine, which is a promising anti-herpes agent also known as H2G. Patent Document 2 discloses 6-deoxy H2G. Patent Document 3 discloses that these compounds, particularly their R-(−) enantiomers, are also active against retroviral infections such as HIV. Various derivatives of H2G, such as phosphate esters, aliphatic esters (eg diacetates and dipropionates) and ethers of hydroxy groups on acyclic side chains are disclosed in US Pat. This patent also describes condensation of acyclic side chains to the N-9 position of typically 6-halogenated purine residues, or alternatively pyrimidine or frazano- [3,4-d] pyrimidine residues. Also disclosed are methods for the preparation of these derivatives, including imidazole ring closures or pyrimidine ring closures of imidazole residues (acyclic side chains already present on the precursor pyrimidine or imidazole residues, respectively). is doing. In the broadest description of each of these methods, the acyclic side chain has been previously derivatized, but individual examples also show one-step diacylation of H2G using acetic anhydride and propionic anhydride and DMF. Yes.

  Harnden et al. (Non-Patent Document 1) describe several short-chain aliphatic esters of 9- [4-hydroxy- (3-hydroxymethyl) butyl] guanine, an acyclic nucleoside also known as pencyclovir and its 6-deoxy analogues were searched. Funciclovir, an antiviral agent on the market, is a diacetyl derivative of 6-deoxypentacyclovir.

  Benjamin et al. (Non-Patent Document 2) disclose short-chain aliphatic esters of 9-[(1,3-dihydroxy-2-propoxy) methyl] guanine, also known as ganciclovir. Dipropionate esters are disclosed as being preferred esters.

  Lake-Bakaar et al., In Non-Patent Document 3, disclose diacetate and dipropionate derivatives of H2G and monoacetate and diacetate derivatives of 6-deoxyH2G. H2G diacetate and dipropionate derivatives have been reported to provide only modest improvements in bioavailability compared to H2G.

  U.S. Patent No. 6,037,097 discloses aliphatic esters of 6-deoxy N-7 analogs of ganciclovir, including di-pivaloyl esters, di-valeroyl esters, mono-valeroyl esters, mono-oleoyl esters and mono-stearoyl esters. Prodrugs are disclosed.

  US Pat. Nos. 5,099,086 and 5,056,086 both disclose mono-ester derivatives of nucleoside analogues derived from mono-unsaturated C18 or C20 fatty acids. U.S. Patent No. 6,057,031 also discloses long chain fatty acid mono-ester derivatives of nucleoside analogues.

  A second approach to providing prodrugs of acyclic nucleosides involves the preparation of amino acid esters of one or more hydroxy groups on the acyclic side chain. Patent Document 8 generally discloses amino acid esters of acyclovir, and Patent Document 9 discloses valine and isoleucine esters of acyclovir.

  U.S. Patent No. 6,057,077 discloses ganciclovir amino acid ester derivatives, including di-valine ester derivatives, di-isoleucine ester derivatives, di-glycine ester derivatives and di-alanine ester derivatives. U.S. Patent No. 6,057,077 discloses penciclovir amino acid ester derivatives, including mono-valine ester derivatives and di-valine ester derivatives.

  Patent Documents 12 and 13 disclose achiral amino acid esters of ganciclovir. U.S. Patent No. 6,057,051 discloses mono-L-valine ester of ganciclovir and its preparation from di-valyl-ganciclovir.

  Patent Document 15 discloses various bisamino acid ester derivatives of 9-[(1 ′, 2′-bishydroxymethyl) -cyclopropan-1′-yl] methylguanine. U.S. Patent No. 6,057,031 discloses aliphatic esters, amino acid esters and mixed acetate / valinate esters of acyclic nucleoside 9- [3,3-dihydroxymethyl-4-hydroxy-but-1-yl] guanine. This document discloses that bioavailability is reduced when one of the valine esters of the trivaline ester derivative is replaced with an acetate ester.

European patent EP165289 European Patent EP186640 Specification European patent EP343133 WO94 / 24134 specification WO93 / 07163 specification WO94 / 22887 specification US Pat. No. 5,216,142 European Patent EP99493 Specification European patent application EP 308065 European patent application EP375329 WO95 / 09855 specification DE19526163 US Pat. No. 5,543,414 European patent application EP694547 European patent application EP 654 473 specification WO95 / 22330 specification

j. Med. Chem. 32, 1738, 1989 Pharm. Res.4, No.2, 120, 1987 Antimicrob. Agents Chemother. 33, No. 1, 110-112, 1989

  The present inventors have found that a diester derivative of H2G having a specific combination of an amino acid ester and a fatty acid ester can significantly improve oral bioavailability compared to the parent compound (H2G).

［式中、
（ａ）Ｒ_１が−Ｃ(Ｏ)ＣＨ(ＣＨ(ＣＨ_３)_２)ＮＨ_２または−Ｃ(Ｏ)ＣＨ(ＣＨ(ＣＨ_３)ＣＨ_２ＣＨ_３)ＮＨ_２であり、Ｒ_２が−Ｃ(Ｏ)Ｃ_３〜Ｃ_２１の飽和もしくはモノ不飽和の任意に置換されたアルキルであるか；または
（ｂ）Ｒ_１が−Ｃ(Ｏ)Ｃ_３〜Ｃ_２１の飽和もしくはモノ不飽和の任意に置換されたアルキルであり、Ｒ_２が−Ｃ(Ｏ)ＣＨ(ＣＨ(ＣＨ_３)_２)ＮＨ_２または−Ｃ(Ｏ)ＣＨ(ＣＨ(ＣＨ_３)ＣＨ_２ＣＨ_３)ＮＨ_２であり；
Ｒ_３はＯＨまたはＨである］
で示される新規な化合物または薬理学的に許容しうるその塩が提供される。 Thus, according to the first aspect of the invention, the formula I :

[Where:
(A) R ₁ is —C (O) CH (CH (CH ₃ ) ₂ ) NH ₂ or —C (O) CH (CH (CH ₃ ) CH ₂ CH ₃ ) NH ₂ , and R ₂ is —C (O) C ₃ -C ₂₁ saturated or monounsaturated optionally substituted alkyl; or (b) R ₁ is —C (O) C ₃ -C ₂₁ saturated or monounsaturated any And R ₂ is —C (O) CH (CH (CH ₃ ) ₂ ) NH ₂ or —C (O) CH (CH (CH ₃ ) CH ₂ CH ₃ ) NH ₂ ;
R ₃ is OH or H]
Or a pharmacologically acceptable salt thereof.

  The beneficial effects on the oral bioavailability of the mixed fatty acids and amino acid esters of the present invention are particularly unexpected compared to the oral bioavailability of the corresponding fatty acid esters. Based on the results using the urine recovery assay of H2G from rats (Table 1A) or plasma drug assay (Table 1B), both the monofatty acid ester and the di-fatty acid ester of H2G were compared to the parent compound H2G for oral bioavailability. There is no improvement. In fact, di-stearate derivatives have significantly lower bioavailability compared to the parent compound, indicating that stearate esters are detrimental in improving the oral bioavailability of H2G. It has been reported that conversion of one or both hydroxyls to the corresponding valine or di-valine ester in certain other acyclic nucleoside analogs improves bioavailability. The same bioavailability improvement was obtained when H2G was converted to the corresponding mono- or di-valyl ester derivative compared to the parent compound. Since H2G fatty acid derivatives have been shown to be detrimental in improving bioavailability, mixed amino acid / fatty acid diester derivatives of H2G are valine diester derivatives of H2G based on urine recovery assay and plasma drug assay, respectively. It was unexpected to confer improved or comparable oral bioavailability compared to.

The present invention also provides a pharmaceutical composition comprising a compound of formula I and a pharmacologically acceptable salt thereof together with a pharmaceutically acceptable carrier or diluent. A further aspect of the present invention is in the use of the compounds of formula I and pharmacologically acceptable salts thereof for the treatment and in the preparation of medicaments for the treatment and prevention of viral infections in humans or animals of these compounds and salts. Including use.

  The compounds of the present invention include herpes infections such as those caused by varicella-zoster herpesvirus, herpes simplex virus types 1 and 2, Epstein-Barr virus, herpes type 6 (HHV-6) and type 8 (HHV-8), among others. It is a powerful antiviral agent against. The compounds of the present invention are particularly useful against varicella-zoster herpesvirus infections such as herpes zoster in older people with post-herpetic neuralgia and chick varicella in younger age, and reduce the duration and severity of the disease for several days Can do. Epstein-Barr virus infections that can be treated using the compounds of the present invention include infectious monocytic hyperplasia / glands that have previously been unable to be treated but cause months of scholastic incapacity in adolescents Contains heat.

  The compounds of the present invention are also active against certain retroviral infections, especially against SIV, HIV-1 and HIV-2 and against infections where transactivating viruses have been shown.

  Accordingly, a further aspect of the present invention provides a method for the prevention or treatment of viral infections in humans or animals comprising administering to humans or animals an effective amount of a compound of formula I or a pharmacologically acceptable salt thereof. To do.

  Advantageously, R3 is hydroxy or its tautomer = O so that, for example, when the side chain is cleaved in vivo, the base moiety of the compounds of the invention is natural guanine. In another embodiment, R3 may be hydrogen and thus define a generally more soluble 6-deoxy derivative that can be oxidized to guanine in vivo by, for example, xanthine oxidase.

The compound of formula I may exist in the form of a racemate that is a mixture of the 2R and 2S isomers. Preferably, however, the compound of formula I has at least 70%, preferably 90% R-form, for example 95% or more R-form. Most preferably, the compound of formula I is the enantiomerically pure R form.

The amino acids of the radicals R ₁ / R ₂ are preferably derived from L-amino acids. The fatty acids of the radicals R ₁ / R ₂ are preferably those having an even number of carbon atoms in total, in particular decanoyl (C ₁₀ ), lauryl (C ₁₂ ), myristoyl (C ₁₄ ), palmitoyl (C ₁₆ ), stearoyl ( C ₁₈ ) or eicosanoyl (C ₂₀ ) is preferred. Other useful groups R ₁ / R ₂ include butyryl, hexanoyl, octanoyl or behenoyl (C ₂₂ ). Further useful groups R ₁ / R ₂ include myristoleic, myristelaidic, palmitoleic acid, palmitelaidic, n6-octadecenoic acid, oleic acid, elaidic acid, gandoic acid And those derived from erucic acid or brassic acid. Monounsaturated fatty acid esters generally have a double bond in the trans configuration, preferably at the ω-6, ω-9 or ω-11 position, depending on their length. Group _R 1 / _{R 2 is,} C 9 _{-C 17} unsaturated, or n: 9 preferably derived from fatty acids containing monounsaturated alkyl.

Saturated or unsaturated fatty acid or _R 1 / _{R 2} is _hydroxy, C 1 -C _{6 alkyl,} C 1 -C _{6 alkoxy,} C 1 -C ₆ alkoxy _C 1 -C _{6 alkyl,} C 1 -C ₆ alkanoyl, amino Optionally substituted with up to 5 same or different substituents independently selected from the group consisting of halo, cyano, azido, oxo, mercapto and nitro.

Most preferred compounds of formula I are those in which R ₁ is —C (O) CH (CH ₃ ) ₂ ) NH ₂ or —C (O) CH (CH (CH ₃ ) CH ₂ CH ₃ ) NH ₂ , and R ₂ There are those saturated alkyl _{-C (O) C 9 ~C 17} .

  As used herein, “lower alkyl” refers to a linear or branched alkyl radical containing 1 to 7 carbon atoms, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec- Examples include but are not limited to butyl, t-butyl, n-pentyl, 1-methylbutyl, 2,2-dimethylbutyl, 2-methylpentyl, 2,2-dimethylpropyl, n-hexyl and the like.

As used herein, “alkanoyl” refers to R ₂₀ C (O) — (wherein R ₂₀ is a lower alkyl group).

As used herein, “alkoxy” refers to R ₂₁ O— (wherein R ₂₁ is a lower alkyl group).

  As used herein, “alkoxyalkyl” refers to an alkoxy group bonded to a lower alkyl radical.

As used herein, “N-protecting group” or “N-protected” refers to protecting the N-terminus of an amino acid or peptide or protecting an amino group from undesired reactions during synthetic procedures. Commonly used N-protecting groups are described in Greene's “Protective Groups in Organic Synthesis” (John Willie and Sons, New York, 1981), which is incorporated herein by reference. . Examples of the N-protecting group include formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, a sheet Le group such as 4-nitrobenzoyl; benzenesulfonyl, sulfonyl groups such as p- toluenesulfonyl, benzyloxycarbonyl, p- chlorobenzyloxycarbonyl, p- methoxybenzyl Oxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxy Rubonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1- (p-biphenylyl) -1-methylethoxycarbonyl, α, α-dimethyl-3,5 -Dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butoxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4- Nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl, etc. Rubamate-forming groups; alkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl; and silyl groups such as trimethylsilyl. Preferred N-protecting groups include formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, phenylsulfonyl, benzyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (Cbz), and the like.

  As used herein, “O-protecting group” or “hydroxy protecting group” or “—OH protecting group” is disclosed in Green's “Protective Groups in Organic Synthesis” (John Willy and Sons, New York, 1981). A substituent that protects the hydroxyl group from undesired reactions during the synthesis procedure, such as an O-protecting group that has been described. O-protecting groups are substituted methyl ethers such as methoxymethyl, benzyloxymethyl, 2-methoxyethoxymethyl, 2- (trimethylsilyl) ethoxymethyl, t-butyl, benzyl and triphenylmethyl: tetrahydropyranyl ether: substituted ethyl ether For example, 2,2,2-trichloroethyl; silyl ethers such as trimethylsilyl, t-butyldimethylsilyl and t-butyldiphenylsilyl: and hydroxyl groups reacted with carboxylic acids such as acetic acid, propionic acid, benzoic acid, etc. The ester prepared by

  In the present specification, the “active ester derivative” refers to an acid halide such as an acid chloride, an anhydride derived from formic acid and acetic acid, an anhydride from an alkoxycarbonyl halide such as isobutyloxycarbonyl chloride, and the like derived from N-hydroxysuccinimide. Ester, ester derived from N-hydroxyphthalimide, ester derived from N-hydroxybenzotriazole, ester derived from N-hydroxy-5-norbornene-2,3-dicarboxamide, ester derived from 2,4,5-trichlorophenyl, sulfone Examples include, but are not limited to, acid-derived anhydrides (for example, p-toluenesulfonic acid-derived anhydride).

Preferred compounds of formula I include the following:
(R) -9- [2- (butyryloxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [2- (4-acetylbutyryloxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [2- (hexanoyloxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2- (octanoyloxymethyl) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2- (decanoyloxymethyl) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2- (dodecanoyloxymethyl) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2- (tetradecanoyloxymethyl) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2- (hexadecanoyloxymethyl) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2- (octadecanoyloxymethyl) butyl] guanine,
(R) -9- [2- (Eicosanoyloxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [2- (docosanoyloxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2-((9-tetradecenoyl) oxymethyl) butyl] guanine,
(R) -9- [2-((9-hexadecenoyl) oxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2-((6-octadecenoyl) oxymethyl) butyl] guanine,
(R) -9- [4- (L-Isoleuciloxy) -2-((9-octadecenoyl) oxymethyl) butyl] guanine,
(R) -9- [2-((11-eicosanoyl) oxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [2-((13-docosenoyl) oxymethyl) -4- (L-isoleuyloxy) butyl] guanine,

(R) -2-amino-9- [2- (butyryloxymethyl) -4- (L-isoleuyloxy) butyl] purine,
(R) -2-amino-9- [2- (4-acetylbutyryloxymethyl) -4- (L-isoleuyloxy) butyl] purine,
(R) -2-amino-9- [2- (hexanoyloxymethyl) -4- (L-isoleuyloxy) butyl] purine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (octanoyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (decanoyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (dodecanoyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (tetradecanoyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (hexadecanoyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (octadecanoyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (eicosanoyloxymethyl) butyl] purine,
(R) -2-amino-9- [2- (eicosanoyloxymethyl) -4- (L-isoleuyloxy) butyl] purine,
(R) -2-amino-9- [2- (docosanoyloxymethyl) -4- (L-isoleuyloxy) butyl] purine,
(R) -2-amino-9- [4- (L-isoleucyloxy) -2-((9-tetradecenoyl) oxymethyl) butyl] purine,
(R) -2-amino-9- [2-((9-hexadecenoyl) oxymethyl) -4- (L-isoleuyloxy) butyl] purine,
(R) -2-amino-9- [4- (L-isoleucyloxy) -2-((6-octadecenoyl) oxymethyl) butyl] purine,
(R) -2-amino-9- [4- (L-isoleucyloxy) -2-((9-octadecenoyl) oxymethyl) butyl] purine,
(R) -2-amino-9- [2-((11-eicosanoyl) oxymethyl) -4- (L-isoleuyloxy) butyl] purine, or (R) -2-amino-9- [2 -((13-docosenoyl) oxymethyl) -4- (L-isoleuyloxy) butyl] purine,
Or a pharmacologically acceptable salt thereof.

More preferred compounds include the following:
(R) -9- [2- (butyryloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (4-acetylbutyryloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (hexanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (octanoyloxymethyl) -4- (L-valyloxy) -butyl] guanine,
(R) -9- [2- (decanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (dodecanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (tetradecanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2-hexadecanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (octadecanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (Eicosanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (Eicosanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2- (docosanoyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2-((9-tetradecenoyl) oxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2-((9-hexadecenoyl) oxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2-((6-Octadecenoyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2-((9-octadecenoyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2-((11-eicosanoyl) oxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -9- [2-((13-docosenoyl) oxymethyl) -4- (L-valyloxy) butyl] guanine,

(R) -2-amino-9- [2- (butyryloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (4-acetylbutyryloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (hexanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (octanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (decanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (dodecanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (tetradecanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (hexadecanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (octadecanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (eicosanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2- (docosanoyloxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2-((9-tetradecenoyl) oxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-Amino-9- [2-((9-hexadecenoyl) oxymethyl)-
4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2-((6-octadecenoyl) oxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2-((9-octadecenoyl) oxymethyl) -4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2-((11-eicosenoyl) oxymethyl) -4- (L-valyloxy) butyl] purine, or (R) -2-amino-9- [2-(( 13-docosenoyl) oxymethyl) -4- (L-valyloxy) butyl] purine,
Or a pharmacologically acceptable salt thereof.

Other preferred compounds of formula I include the following:
(R) -9- [4- (butyryloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (4-acetylbutyryloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (hexanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (octanoyloxy) -2- (L-valyloxymethyl) -butyl] guanine,
(R) -9- [4- (decanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (dodecanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (tetradecanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (hexadecanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (octadecanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (Eicosanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4- (Docosanoyloxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4-((9-tetradecenoyl) oxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4-((9-hexadecenoyl) oxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4-((6-Octadecenoyl) oxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4-((9-octadecenoyl) oxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4-((11-eicosenoyl) oxy) -2- (L-valyloxymethyl) butyl] guanine,
(R) -9- [4-((13-docosenoyl) oxy) -2- (L-valyloxymethyl) butyl] guanine,

(R) -2-amino-9- [4- (butyryloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (4-acetylbutyryloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (hexanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (octanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (decanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (dodecanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (tetradecanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (hexadecanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (octadecanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (eicosanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4- (docosanoyloxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4-((9-tetradecenoyl) oxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4-((9-hexadecenoyl) oxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4-((6-octadecenoyl) oxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4-((9-octadecenoyl) oxy) -2- (L-valyloxymethyl) butyl] purine,
(R) -2-amino-9- [4-((11-eicosanoyl) oxy) -2- (L-valyloxy) butyl] purine, or (R) -2-amino-9- [2-((13 -Docosenoyl) oxymethyl) -2- (L-valyloxy) butyl] purine,
Or a pharmacologically acceptable salt thereof.

The compound of formula I forms a salt, which forms another aspect of the present invention. Suitable pharmacologically acceptable salts of the compounds of formula I include salts of organic acids, especially carboxylic acids, such as acetic acid, trifluoroacetic acid, lactic acid, gluconic acid, citric acid, tartaric acid, maleic acid. , Malic acid, pantothenic acid, isethionic acid, adipic acid, alginic acid, aspartic acid, benzoic acid, butyric acid, digluconic acid, cyclopentanoic acid, glucoheptanoic acid, glycerophosphoric acid, succinic acid, heptanoic acid, hexanoic acid, fumaric acid, nicotinic acid Including salts of palmitic acid (palmoate), pectinic acid, 3-phenylpropionic acid, picric acid, pivalic acid, propionic acid, tartaric acid, lactobionic acid, pivorate, camphoric acid, undecanoic acid and succinic acid Organic sulfonic acid salts such as methanesulfonic acid, ethanesulfone, but not limited to Acids, 2-hydroxyethanesulfonic acid, camphorsulfonic acid, 2-naphthalenesulfonic acid, benzenesulfonic acid, p-chlorobenzenesulfonic acid and p-toluenesulfonic acid salts; and inorganic acid salts such as hydrochloric acid, hydrobromic acid And salts of hydroiodic acid, sulfuric acid, bisulfuric acid, hemisulfate, thiocyanic acid, persulfuric acid, phosphoric acid and sulfonic acid. The hydrochloride is convenient.

The compound of formula I can be isolated as a hydrate. The compounds of the invention can be isolated in crystalline form, preferably in homogeneous crystalline form, so other aspects of the invention are> 70%, preferably> 90% homogeneous crystalline material, eg> 95% homogeneous crystals A compound of formula I is provided in substantially pure crystalline form comprising the substance.

The compounds of the invention are particularly suitable for oral administration but may be administered rectally, vaginally, nasally, topically, transdermally or parenterally, eg intramuscularly, intravenously or epidurally. You can also. The compounds of the invention can be administered alone, for example, in a capsule, but will generally be administered with a pharmaceutically acceptable carrier or diluent. The invention also relates to a process for the preparation of a pharmaceutical composition, which comprises mixing a compound of formula I or a pharmacologically acceptable salt thereof with a pharmacologically acceptable carrier or diluent.

  Oral dosage forms are prepared as unit dosage forms such as capsules and tablets using conventional carriers or binders such as magnesium stearate, milk, starch, lactose, wax, gum or gelatin. Liposomes or synthetic or natural polymers such as HPMC and PVP can be used to make sustained release formulations. Alternatively, the formulations are also provided as nasal or eye drops, syrups, gels or creams, such as water, saline, ethanol, vegetable oils or glycerin, optionally with perfumes and / or preservatives and / or emulsifiers. Includes solutions, suspensions, emulsions, oil-in-water or water-in-oil formulations in conventional vehicles.

  The compounds of the invention are generally in the range of 0.1 to 200 mg / kg / day, advantageously 0.5 to 100 mg / kg / day, more preferably 10 to 50 mg / kg / day, for example 10 to 25 mg / kg. Can be administered as a daily dose within the range of / day. Typical dosages for normal adults are about 50-500 mg, for example 300 mg, once or twice a day for herpes infections and 2-10 times that amount for HIV infections.

  As cautious in antiviral therapy, the compounds of the present invention may be used with other antiviral agents such as acyclovir, valcyclovir, penciclovir, fanciclovir, ganciclovir and its prodrugs, cidofovir, phoscarnet, etc. for herpes indications. And for retroviral indications, it can be administered with AZT, ddI, ddC, d4T, 3TC, Foscarnet, Ritonavir, Indinavir, Saquinavir, Delaviridine, Vertex VX478, Agron AG1343, and the like.

  The compounds of the present invention can be prepared fresh, i.e. by esterification of the H2G parent compound, which has been prepared by the synthetic method disclosed in European Patent Application EP 343133 (cited herein for reference). The

A typical reaction scheme for the preparation of H2G is shown below:

The condensation reaction in step 1 is generally performed in a solvent such as DMF using a base catalyst such as NaOH or Na ₂ CO ₃ . Step 2 includes a reduction process, which can be performed using LiBH ₄ / tetrahydrofuran in a solvent such as t-BuOH. The substitution of chlorine with an amino group in step 3 can be performed under ammonia pressure. Step 4 uses adenosine deaminase (which can be conveniently immobilized on a solid support). When the reaction mixture is cooled, the unreacted isomer precursor remains in solution, thereby increasing the purity.

The starting materials of the compounds of the invention where R ₃ is hydrogen can be prepared by the method shown in European Patent EP 186640, the contents of which are incorporated herein by reference. These starting materials are optionally protected after purifying the 2-amino group of purine with the usual N-protecting groups mentioned above, in particular BOC (t-BuO-CO-), Z (BnO-CO-) or Ph ₃ C- Acylation can be done as described for H2G below.

The compounds of the present invention can be prepared from H2G as described in Schemes A and B below.
A. Direct acylation method

Scheme A is shows the preparation of compounds wherein R ₁ is derived from the origin and R ₂ are fatty acids to amino acids, the reverse reaction scheme is applicable to compounds where R ₂ is derived from R ₁ is a fatty acid derived from the amino acid. In the variables clearly shown in Scheme A above, G is guanine or 6-deoxyguanine, PG is any N-protecting group or hydrogen, R ₁ ^* is a valine or isoleucine side chain, and R ₂ ^* Is a fatty acid side chain. H2G is depicted as starting materials in the above, this compound (not shown), of course at the 2-position of the R ₃ or purine may be protected optionally by conventional N- protecting group. H2G (derivative) reacts in the first step with an activated R ₁ α-amino acid derivative (described further below) in a solvent such as dimethylformamide or pyridine to give a monoacylated product. The R ₁ α-amino acid may be appropriately N-protected with N-BOC or N-CBz. This first acylation can occur preferentially at the side chain 4-hydroxy group on the side chain of H2G under controlled conditions. These controlled conditions can be achieved, for example, by manipulating, inter alia, the reagent concentration and addition rate of the acylating agent, by lowering the temperature, or by selecting a solvent. The reaction can be followed by TLC and the control conditions can be monitored.

After purification, using the same procedure as in the first esterification step, the R ₁ monoacylated compound is further acylated with a suitable activated fatty acid derivative on the side chain 2-CH ₂ OH group to give the diacylated product. The diester product is then subjected to conventional deprotection using, for example, trifluoroacetic acid, HCl (aqueous) / dioxane, or hydrogenation in the presence of a catalyst to give the desired compound of formula I. This compound may be in the form of a salt depending on the deprotection conditions.

Activated R ₁ / R ₂ acid derivatives used for various acylations include, for example, acid halides, acid anhydrides, activated acid esters, or acids in the presence of a coupling agent such as dicyclohexylcarbodiimide, In each case, the “acid” in each represents the corresponding R ₁ / R ₂ amino acid or R ₁ / R ₂ fatty acid. Typical activated acid derivatives include acid chloride, mixed anhydrides derived from formic acid and acetic acid, anhydrides derived from alkoxycarbonyl halides such as isobutyloxycarbonyl chloride, esters derived from N-hydroxysuccinamide, N-hydroxyphthalimide Ester derived from, ester derived from N-hydroxy-5-norbornene-2,3-dicarboxamide, ester derived from 2,4,5-trichlorophenol, anhydride derived from sulfonic acid (for example, anhydride derived from p-toluenesulfonic acid) Etc.).

（式中、Ｇ、ＰＧ、Ｒ_１ ^＊およびＲ_２ ^＊は反応式Ａと同じ） B. Acylation via protection of the side chain 4-hydroxy group:

(In the formula, G, PG, R ₁ ^* and R ₂ ^* are the same as in Reaction Formula A)

Reaction Formula B is illustrated to show the preparation of compounds where R ₁ is derived from an amino acid and R ₂ is derived from a fatty acid, but the reverse reaction formula applies to compounds where R ₁ is derived from a fatty acid and R ₂ is derived from an amino acid. It will be possible. This reaction formula relies on regioselective protection of the H2G side chain 4-hydroxy group with a bulky protecting group. In Reaction Scheme B above, this was shown as t-butyldiphenylsilyl, but trityl, 9- (9-phenyl) xanthenyl, 1,1-bis (4-methylphenyl) -1′-pyrenylmethyl is also suitable. Yes. The resulting product is acylated with a side chain 2-hydroxymethyl group using reagents and procedures similar to those described in Scheme A above, in which case the activated acid derivative is an R ₂ fatty acid such as myristin. Acid, stearic acid, oleic acid, elaidic acid chloride and the like. Thus, the monoacylated compound is subjected to an appropriate deprotection treatment to remove the side chain 4-hydroxy protecting group, which depends on a regioselective protecting group such as HF / pyridine, manipulation of the above reaction conditions, That is, it can be performed in a highly selective manner using reagent concentration, addition rate, temperature and solvent. Subsequently, the released side chain 4-hydroxy group is acylated with an activated α-amino acid in the same manner as described in the above reaction formula A.

Other methods for introducing the amino acid ester of R ₁ / R _{2 in} the reaction formulas A, B, C, D or E in the present specification include, for example, 2-oxa described in International Patent Application No. WO 94/29311. Examples include the -4-azacycloalkane-1,3-dione method.

Other methods for introducing R ₁ / R ₂ fatty acid esters, for example in Scheme A, B, C, D or E herein, include SP435 immobilized Candida antarcticus (Novo・ Preparative Biotransformations using a lipase such as Nordisk), porcine pancreatic lipase or Candida rugosa lipase, 1.11.8 (edited by Roberts) And the enzymatic pathway described in Willy and Sons, New York, 1995). Enzymatic acylation is particularly advantageous when it is desirable to avoid N-protection and deprotection steps on other acyl groups or purines on the 2-amine.

Another route to compounds wherein R ₃ is hydrogen in formula I, the amino acid ester residue of the corresponding guanine compound of formula I (R 1 _/ R ₂ is in optionally protected in the usual N- protecting group such as BOC 6) with an activator such as halo. The 6-purine thus activated is then reduced to purine, for example with a palladium catalyst, and deprotected to the desired 6-deoxy H2G di-ester.

Thus, another aspect of the present invention is
(A) optionally N-protecting purine positions 2 and / or 6 of a compound in which R ₁ and R ₂ are each hydrogen in formula I ;
(B) (i) an optionally N-protected valine or isoleucine group,
(Ii) an optionally substituted saturated or monounsaturated C ₃ -C ₂₁ COOH derivative, or (iii) a regioselective selection of a compound of formula I with a side chain 4-hydroxy group Acylated,
(C) acylating side chain 2-hydroxymethyl groups with (i) optionally N-protected valine or isoleucine derivatives, or (ii) optionally substituted saturated or monounsaturated C ₃ -C ₂₁ COOH derivatives;
(D) if a regioselective protecting group is present at R _1, it is either (i) optionally N-protected valine or isoleucine group, or (ii) optionally substituted saturated or monounsaturated C _A process for the preparation of a compound of formula I is provided, comprising substitution with a _{3 to} C ₂₁ COOH derivative and then (e) deprotecting the resulting compound if necessary.

In the above reaction formulas A and B, selective acylation in which amino acid esters and fatty acid esters are added stepwise is employed. Another method for the preparation of compounds of formula I starts with a diacylated H2G derivative in which both acyl groups are the same, selectively removing one of these acyl groups to give a monoacyl intermediate, followed by a second Different acyl groups are acylated in the same manner as in the above reaction formulas A and B.

Accordingly, another aspect of the present invention provides a process for the preparation of a compound of formula I above,
(A) In Formula I , R ₁ and R ₂ are both valyl or isoleucyl ester (optionally N-protected) or R ₁ and R ₂ are both —C (═O) C ₃ -C Monoacylated diacyl compounds that are _21- saturated or monounsaturated optionally substituted alkyls, and then (B) thus free side-chain 4-hydroxy or side-chain 2-hydroxymethyl groups to the corresponding valyl, acylated with isoleucyl or _{-C (= O) C 3 ~C} 21 saturated or optionally substituted alkyl monounsaturated consists of deprotecting if then (C) required.

  This alternative method is advantageous in that it facilitates the preparation of diacylated H2G derivatives and requires little or no purification steps. Selective removal of only one of the acyl groups of the diacylated H2G derivative can be achieved by manipulating the reaction conditions, particularly temperature, the rate of addition of the reactants and the selection of the base.

Therefore, compounds that can be used in this alternative synthetic route have the formula:

Wherein R ₁ and R ₂ are valyl or isoleucyl (optionally N-protected) or —C (═O) C ₃ -C ₂₁ saturated or monounsaturated optionally substituted alkyl; R ₃ Is OH or H)
It is what has.

In order to facilitate synthesis in this alternative method, R ₁ and R ₂ are preferably the same from the beginning, and most preferably the same amino acid ester. Such di-amino acid esters are generally N-protected in their preparation and can be used directly in the selective deacylation step under these conditions. Alternatively, such N-protected di-aminoacylated H2G derivatives can be deprotected and reprotected as necessary, as described below. Therefore, unprotected di-aminoacyl H2G derivatives include one of the following compounds:
(R) -9- [2- (L-Isoleuciloxymethyl) -4- (L-Isoleuciloxy) butyl] guanine,
(R) -9- [2- (L-valyloxymethyl) -4- (L-valyloxy) butyl] guanine,
(R) -2-amino-9- [4- (L-isoleuyloxy) -2- (L-isoleuyloxymethyl) butyl] purine, and (R) -2-amino-9- [4 -(L-valyloxy) -2- (L-valyloxymethyl) butyl] purine.

  These unprotected H2G diacylated derivatives can be directly subjected to selective deacylation of one of the acyl groups (typically the side chain 4-position acyl) and then the free 4-hydroxy is as above. To enzymatically acylate. Alternatively, the unprotected H2G diacylated derivative is reprotected and then subjected to selective deacylation followed by normal acylation with a fatty acid ester as described in Schemes A and B. be able to. Conveniently, such a reprotection step is performed with a different N-protecting group having properties suitable for subsequent acylation. For example, when preparing di-amino acid H2G derivatives, it is convenient to use a lipophilic N-protecting group such as Fmoc because the lipophilic nature of the protecting group helps to separate the acylated product. . On the other hand, the lipophilic nature of Fmoc has little utility when acylating with fatty acids, so it is convenient to reprotect diacylated H2G with another N-protecting group such as BOC. Good.

（式中、Ｒ_１およびＲ_２の一方は
（i）−Ｃ(Ｏ)ＣＨ(ＣＨ(ＣＨ_３)_２)ＮＨ_２または−Ｃ(Ｏ)ＣＨ(ＣＨ(ＣＨ_３)ＣＨ_２ＣＨ_３)ＮＨ_２、
（ii）−Ｃ(＝Ｏ)Ｃ_３〜Ｃ_２１飽和またはモノ不飽和の任意に置換されたアルキル、または
（iii）位置選択的な保護基;
Ｒ_１およびＲ_２の他方は水素;
Ｒ_３はＯＨまたはＨ）
で示されるものである。 It will also be apparent that the preparation of the compound of formula I can be started from the novel monoacylated intermediate of step (b) (i), (ii) or (iii) in the process of the first aspect of the invention above. Therefore, these compounds have the formula:

Wherein _one of R ₁ and R ₂ is (i) —C (O) CH (CH (CH ₃ ) ₂ ) NH ₂ or —C (O) CH (CH (CH ₃ ) CH ₂ CH ₃ ) NH ₂ ,
(Ii) -C (= O) C 3 ~C 21 saturated or optionally substituted alkyl monounsaturated or (iii) regioselective protecting group;
The other of R ₁ and R ₂ is hydrogen;
R ₃ is OH or H)
It is shown by.

Therefore, useful compounds include the following:
(R) -9- [2-hydroxymethyl-4- (t-butyldiphenylsilyl) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (trityloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (9- (9-phenyl) xanthenyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (1,1-bis (4-methylphenyl) -1′-pyrenylmethyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (decanoyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl) -4- (dodecanoyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (tetradecanoyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl) -4- (hexadecanoyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (octadecanoyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl) -4- (eicosanoyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (docosanoyloxy) butyl] guanine,
(R) -9- [4-hydroxy-2- (decanoyloxymethyl) butyl] guanine,
(R) -9- [4-hydroxy-2- (dodecanoyloxymethyl) butyl] guanine,
(R) -9- [4-hydroxy-2- (tetradecanoyloxymethyl) butyl] guanine,
(R) -9- [4-hydroxy-2- (hexadecanoyloxymethyl) butyl] guanine,
(R) -9- [4-hydroxy-2- (octadecanoyloxymethyl) butyl] guanine,
(R) -9- [4-hydroxy-2- (eicosanoyloxymethyl) butyl] guanine,
(R) -9- [4-hydroxy-2- (docosanoyloxymethyl) butyl] guanine,
(R) -9- [2-hydroxymethyl-4- (L-valyloxy) butyl] guanine,
(R) -9- [2-hydroxymethyl) -4- (L-isoleuyloxy) butyl] guanine,
(R) -9- [4-hydroxy-2- (L-isoleucyloxymethyl) butyl] guanine,
(R) -9- [4-hydroxy-2- (L-valyloxymethyl) butyl] guanine,
(R) -2-amino-9- [2-hydroxymethyl-4- (L-valyloxy) butyl] purine,
(R) -2-amino-9- [2-hydroxymethyl) -4- (L-isoleuyloxy) butyl] purine,
(R) -2-amino-9- [4-hydroxy-2- (L-isoleuyloxymethyl) butyl] purine and (R) -2-amino-9- [4-hydroxy-2- (L- Valyloxymethyl)
Butyl] purine.

The regioselectively protected side chain 4-hydroxy intermediate in step (c) of the method of the first aspect of the present invention is also a novel compound.
Therefore, useful compounds include the following:
(R) -9- [2-decanoyloxymethyl-4- (t-butyldiphenylsilyl) butyl] guanine,
(R) -9- [2-dodecanoyloxymethyl-4- (t-butyldiphenylsilyl) butyl] guanine,
(R) -9- [2-tetradecanoyloxymethyl-4- (t-butyldiphenylsilyl) butyl] guanine,
(R) -9- [2-hexadecanoyloxymethyl-4- (t-butyldiphenylsilyl) butyl] guanine,
(R) -9- [2-octadecanoyloxymethyl-4- (t-butyldiphenylsilyl) butyl] guanine,
(R) -9- [2-eicosanoyloxymethyl-4- (t-butyldiphenylsilyl) butyl] guanine, and (R) -9- [2-docosanoyloxymethyl-4- (t-butyldiphenyl) Silyl) butyl] guanine.

Another method for preparing compounds of the present invention where R ₃ is —OH in Formula I is shown in Reaction Scheme C.

Referring to Scheme C, malonic acid ester 1 (R ₄ and R ₅ are lower alkyl or benzyl, etc.) is treated with an inert solvent (eg, DMF or THF or dioxane or dioxolane) at a temperature of about −40 ° C. to about 190 ° C. Or about 0.5 to about 2.0 molar equivalents of a base (such as potassium t-butoxide or sodium ethoxide or NaH or KH) in N-methylpyrrolidone and the like). 0.0 molar equivalent of acetal 2 (R ₆ and R ₇ are lower alkyl or benzyl, or R ₆ and R ₇ together are —CH ₂ CH ₂ — or —CH ₂ CH ₂ CH ₂ — or —CH ₂ CH ₂ CH ₂ CH ₂ —, X ₁ is a leaving group (eg Cl, Br or I, or methanesulfonate, triflate , P-toluenesulfonate, sulfonate such as benzenesulfonate) and the like to obtain alkylated malonic acid ester 3 . The alkylated malonate 3 can be purified by distillation or by treating the crude alkylated malonate with a dilute base solution (eg 7% aqueous KOH) and then removing volatile impurities by distillation. Can do.

3 to about 0.5 to about 4.0 molar equivalents of ester → alcohol reduction in an inert solvent (such as THF or methyl t-butyl ether or t-BuOH) at a temperature of about −20 ° C. to about 100 ° C. Reduction with an agent (eg, LiBH ₄ or Ca (BH ₄ ) ₂ or NaBH ₄ or LiAlH ₄ ) yields diol 4 . From about 1.0 to about 20.0 molar equivalents of vinyl ester 5 (R ₈ is C Enzymatic esterification of 4 by reaction with _{3 to} C ₂₁ saturated or monounsaturated optionally substituted alkyl) provides the desired stereoisomer of ester 6 . This reaction can be carried out in the absence of a solvent or in the presence of an inert solvent such as methyl t-butyl ether or toluene or hexane. This reaction is carried out at a temperature of about -20 ° C to about 80 ° C.

6 in halogenated agents (eg NBS / P (Ph) ₃ or NCS / P (Ph) in an inert solvent such as methylene chloride or toluene or ethyl acetate at a temperature of about −25 ° C. to about 100 ° C. ) ₃ or POCl ₃ or NCS / P (Ph) ₃ / NaI in acetone) or in an inert solvent (such as methylene chloride or toluene or ethyl acetate) for about 1.0 to about 4. About 0.8 to about 2.0 molar equivalents of a sulfonyl halide (eg, benzenesulfonyl chloride, toluene, etc.) in the presence of 0 molar equivalents of a base (eg, triethylamine or potassium carbonate or pyridine or dimethylaminopyridine or ethyldiisopropylamine). Sulfonyl chloride or methanesulfonyl chloride) By reacting, leaving the alcohol substituent of 6 (e.g., halogen or sulfonate) ester 7 was converted to (X ₂ is a halogen or sulfonate leaving group) obtained.

7 at about −25 ° C. to about 140 ° C. in an inert solvent such as DMF or THF or acetonitrile or N-methylpyrrolidone or ethanol. give the substituted purine 9 is reacted with the presence of about 0.9 to about 2.0 molar equivalents under 2-amino-4-chloropurine 8 such as potassium carbonate or NaH or KH or NaOH or KOH or lithium diisopropylamide) .

Alternatively, alcohol 6 is Mitsunobu coupled with 2-amino-4-chloropurine 8 (eg, P (Ph) ₃ / diethyl azidocarboxylate) 9 .

9 to about 1.0 to about 6.0 molar equivalents of a base (eg, potassium t-butoxide or potassium carbonate in an inert solvent (eg, THF or DMF) at a temperature of about −25 ° C. to about 150 ° C. or NaH or KH or alcohol _R 9 OH in about 2.0 to about 20 molar equivalents in the presence of lithium diisopropylamide, etc.) _{(R 9} is to obtain the alcohol 10 is reacted with an alcohol protecting group) such as benzyl.

10 alcohol protecting groups R ₉ are removed (eg catalytic hydrogen in an inert solvent such as ethanol, benzyl alcohol, methanol or THF in the presence of a hydrogenation catalyst such as Pd / C or Pd (OH) ₂ To obtain substituted guanine 11 .

11 at a temperature of about −25 ° C. to about 100 ° C. (a) about 0.8 to about 2.0 molar equivalents of R ₁₀ COOH and a coupling reagent (in an inert solvent such as THF or DMF) ( For example, DCC / DMAP, etc.) or (b) from about 0 to about 3.0 molar equivalents of base (eg, methylene chloride or THF or pyridine or acetonitrile or DMF, etc.) From about 0.8 to about 2.0 molar equivalents of an activated derivative of R ₁₀ COOH in the presence of pyridine or dimethylaminopyridine or triethylamine or ethyldiisopropylamine or N-methylmorpholine or DBU or potassium carbonate (eg, acid chloride or N- hydroxysuccinimide ester or _{R 10} C (O) OS (O) ₂ R ₃₀ (wherein R ₃₀ is lower alkyl, phenyl or toluyl) or R ₁₀ C (O) OC (O) R ₁₀ or R ₁₀ C (O) OC (O) R _10a (wherein , R _10a is esterified by reacting with lower alkyl etc.) to give ester 12 . R ₁₀ is C ₃ -C ₂₁ saturated or monounsaturated optionally substituted alkyl.

12 in an inert solvent (eg, THF / H ₂ O or methylene chloride / H ₂ O or ethyl acetate / H ₂ O or ethanol / H ₂ O or methanol / H ₂ O). The 12 acetal substituents are deprotected by reaction with 10.0 molar equivalents of an acid (such as trifluoromethanesulfonic acid or HCl or acetic acid or sulfuric acid) to reduce the resulting aldehyde. The resulting crude reaction mixture is charged with about 0.1 to about 10.0 molar equivalents of a base (such as sodium or potassium bicarbonate or triethylamine or pyridine or KOH) (optionally further with an inert solvent (for example THF)). And / or methylene chloride or ethyl acetate or methyl t-butyl ether or isopropanol)) and about 0.3 to about 5.0 molar equivalents of an aldehyde reducing agent (eg, sodium borohydride or RaNi / H ₂ or borane t) -Butylamine complex etc.) is added at a temperature of about -25 ° C to about 100 ° C to give alcohol 13 . The optical purity of compound 13 can be increased by reacting with an optically active organic sulfonic acid such as (S)-(+)-camphorsulfonic acid. The preferred sulfonic acid for this purpose is (S)-(+)-camphor sulfonic acid.

Alternatively, the 12 acetal substituents can be hydrolyzed by reacting with an acidic resin (eg, Amberlyst 15 resin, Nafion NR50 resin, Dowex 50WX4-200R resin or Amerlite 120 resin) in an inert solvent. The aldehyde can be obtained. The aldehyde can be isolated prior to reduction to alcohol 13 as described above, or the crude aldehyde can be reduced in situ.

13 in an inert solvent such as THF or dioxane or dioxolane or DMF or methylene chloride at a temperature of about 25 ° C. to about 100 ° C. Reaction with ₁ NHCH (R ₁₁ ) COOH or an activated derivative thereof (P ₁ is an N-protecting group, R ₁₁ is isopropyl or isobutyl) provides alcohol 14 . 14 is N-deprotected to give a compound of the invention where R ₃ is —OH in Formula I.

Alternatively, compound 13 can be converted to a symmetric anhydride derived from P ₁ NHCH (R ₁₁ ) COOH (ie, P ₁ NHCH (R ₁₁ ) C (O) O—C (O) CH (R ₁₁ ) NHP ₁ ). Reaction 14 can be obtained. The anhydride can be prepared in situ or it can be prepared separately before reacting with 13 .

Alternatively, 11 hydrolyzes the ester of 9 to an alcohol (eg, by reacting with K ₂ CO ₃ in MeOH / H ₂ O) and then converts the chloro group directly to —OH ( For example, by reacting with an inorganic base such as KOH or NaOH in H ₂ O under heating).

As yet another alternative, 11 can be prepared directly by hydrolysis of the chloro-ester 9 (eg, by reacting with an inorganic base such as KOH or NaOH in H ₂ O under heating).

As yet another alternative, 11 is from 9 (or from a hydroxy compound resulting from hydrolysis of the ester in 9 ), an inorganic base in an aqueous solvent (eg, NaOH, LiOH, KOH, etc., especially NaOH) and trimethylamine Can be prepared by reacting with.

As yet another alternative, 11 is the presence of mercaptoethanol in the hydrolysis of chloro-ester 9 (eg, in a mixed solvent of water and methanol or dioxane, etc.) at a temperature from about 20 ° C. to near the reflux temperature. For example, by reacting with 1 to 3 equivalents of sodium methoxide base, etc.).

As yet another alternative, 13 can be prepared by reacting 9 (wherein R ₈ = R ₁₀ ) with formic acid optionally by heating and then reducing the aldehyde to give 13 .

Another method for preparing compounds in which R ₃ is —OH in Formula I is shown in Reaction Formula D.

Malonic acid ester 1 (R ₄ and R ₅ are, for example, lower alkyl or benzyl) in an inert solvent (eg, DMF or THF or dioxane or dioxolane or N-methylpyrrolidinone) at a temperature of about −40 ° C. to about 190 ° C. In the presence of about 0.5 to about 2.0 molar equivalents of a base (such as potassium t-butoxide or sodium ethoxide or NaH or KH) in the presence of about 0.5 to about 2.0 molar equivalents of ether 15 (X ₁ is a leaving group (for example, Cl, Br or I, or sulfonates such as methanesulfonate, triflate, p-toluenesulfonate, benzenesulfonate, etc.), R ₁₂ is —CH (Ph) ₂ , —C (Ph) _3. Or alkylated malonate ester 16 by reaction with -Si (t-Bu) (Me) ₂ or the like) Get.

16 at a temperature of about −20 ° C. to about 100 ° C. in an inert solvent such as THF or methyl t-butyl ether or ethanol or t-butanol. Reduction with an alcohol reducing agent (such as LiBH ₄ or Ca (BH ₄ ) ₂ or NaBH ₄ or LiAlH ₄ ) yields diol 17 . About 1.0 to about 20.0 molar equivalents of vinyl ester 5 (R ₈ is C ₈ ) in the presence of lipase (eg, lipase PS-30 or lipase PPL or lipase CCL) or phospholipase (eg, phospholipase D). Enzymatic esterification of 17 by reaction with _3- C ₂₁ saturated or monounsaturated optionally substituted alkyl) provides the desired stereoisomer of ester 18 . This reaction can be carried out in the absence of a solvent or in the presence of an inert solvent such as methyl t-butyl ether or toluene or hexane. This reaction is carried out at a temperature of about -20 ° C to about 80 ° C.

18 in halogenated agents (eg NBS / P (Ph) ₃ or NCS / P (Ph) in an inert solvent such as methylene chloride or toluene or ethyl acetate at a temperature of about −25 ° C. to about 100 ° ) ₃ or POCl ₃ or NCS / P (Ph) ₃ / NaI in acetone) or about 1.0 to about 4.0 molar equivalents of a base (eg, triethylamine or potassium carbonate or pyridine or methyl t 18 alcohol substituents by reacting with about 0.8 to about 2.0 molar equivalents of a sulfonyl halide (eg, benzenesulfonyl chloride, toluenesulfonyl chloride or methanesulfonyl chloride) in the presence of Converted to a leaving group (eg halogen or sulfonate) Ter 19 (X ₂ is a halogen or sulfonate leaving group) is obtained.

19 in an inert solvent (such as DMF or THF or acetonitrile or N-methylpyrrolidone or ethanol) at a temperature of about −25 ° C. to about 140 ° C. (E.g., potassium carbonate or NaH or KH or NaOH or KOH or lithium diisopropylamide) in the presence of about 0.9 to about 2.0 molar equivalents of 2-amino-4-chloropurine 8 to give substituted purine 20 Get.
Alternatively, alcohol 18 is Mitsunobu coupled with 2-amino-4-chloropurine 8 (eg, P (Ph) ₃ / diethyl azidocarboxylate) 20 .

20 to about 1.0 to about 6.0 molar equivalents of a base (eg, potassium t-butoxide or potassium carbonate) in an inert solvent (eg, THF or DMF) at a temperature of about −25 ° C. to about 150 ° C. or NaH or KH or lithium diisopropylamide, etc.) alcohol _R 9 OH from about 2.0 to about 20.0 molar equivalents in the presence of _{(R 9} is to obtain the alcohol 21 is reacted with an alcohol protecting group) such as benzyl.

Removing the alcohol protecting group R ₉ of 21 (eg, catalytic hydrogen in an inert solvent such as ethanol, benzyl alcohol, methanol or THF in the presence of a hydrogenation catalyst such as Pd / C or Pd (OH) ₂ To obtain substituted guanine 22 . 22 can be esterified as described in Scheme C to give 23 .

23 is reacted with (a) a reducing agent such as HCO ₂ H and Pd / C (when R ₁₂ is —CH (Ph) ₂ or —C (Ph) ₃ ), or (b) desilylation The ether substituent of 23 is deprotected to give 13 by reacting with an agent such as Bu ₄ NF (when R ₁₂ is —Si (t-Bu) (Me) ₂ ). Alcohol 13 can be converted to 1 as shown in Reaction Scheme C.

Alternatively, 22 can hydrolyze the 20 ester to an alcohol (eg, by reacting with K ₂ CO ₃ in MeOH / H ₂ O) and then convert the chloro group directly to —OH ( For example, by reacting with an inorganic base such as KOH or NaOH in H ₂ O under heating).

As yet another alternative, 22 can be prepared directly by hydrolysis of chloro-ester 20 (eg, by reacting with KOH in H ₂ O under heating).

As yet another alternative, 22 is from 20 (or from a hydroxy compound resulting from hydrolysis of the ester in 20 ), an inorganic base (eg, NaOH, LiOH, KOH, etc., preferably NaOH) in an aqueous solvent and It can be prepared by reacting with trimethylamine.

As yet another alternative, 22 is the presence of mercaptoethanol in the hydrolysis of chloro-ester 20 (eg, in a mixed solvent of water and methanol or dioxane, etc.) at a temperature from about 20 ° C. to near reflux temperature. For example, by reacting with 1 to 3 equivalents of sodium methoxide base, etc.).

As yet another alternative, 23 can be prepared by reacting 20 (wherein R ₈ = R ₁₀ ) with formic acid, optionally with heating, and then reducing the aldehyde to obtain 23 .

Yet another approach is to enzymatically esterify alcohol 4 or 17 with vinyl ester CH ₂ ═CH—OC (O) R ₁₀ (ie, R ₈ = R ₁₀ in Schemes C and D) in 6 or 18 Directly introducing the desired carboxylic acid ester of the final product I. Thus, it is possible to dispense with the ester hydrolysis and re-esterification contained in 9-12 or 20-23.

  The procedures of Schemes C and D are characterized by the fact that each hydroxyl group of the acyclic side chain is distinguished by the use of a different hydroxy protecting group or precursor group. This allows for selective acylation of each hydroxy group with either an amino acid group or a fatty acid group.

Reaction formulas C and D are described and described with reference to embodiments of the compounds of the invention in which R ₁ is derived from an amino acid and R ₂ is derived from a fatty acid. However, it will be apparent that each opposite reaction scheme can be applied to compounds where R ₁ is derived from a fatty acid and R ₂ is derived from an amino acid.

Yet another method for preparing compounds of formula I is shown in Scheme E. About 1.0 to about 20.0 molar equivalents of vinyl ester 24 (R ₁₀ is C ₁₀ ) in the presence of lipase (eg, lipase PS-30 or lipase PPL or lipase CCL) or phospholipase (eg, phospholipase D). Enzymatic esterification of 4 (see Scheme C) by reaction with _{3 to} C ₂₁ saturated or monounsaturated optionally substituted alkyl) provides the desired stereoisomer of ester 25 . This reaction can be carried out in the absence of a solvent or in the presence of an inert solvent such as methyl t-butyl ether or toluene or hexane. This reaction is carried out at a temperature of about -20 ° C to about 80 ° C.

25 in a inert solvent such as methylene chloride or toluene or ethyl acetate at a temperature of about −25 ° C. to about 100 ° C. (eg, NBS / P (Ph) ₃ or NCS / P (Ph ) ₃ or POCl ₃ or NCS / P (Ph) ₃ / NaI in acetone) or about 1 in an inert solvent (such as methylene chloride or toluene or ethyl acetate or pyridine or methyl t-butyl ether). About 0.8 to about 2.0 molar equivalents of a sulfonyl halide (e.g., triethylamine or potassium carbonate or pyridine or dimethylaminopyridine or ethyldiisopropylamine). , Benzenesulfonyl chloride, toluenesulfonyl chloride Others by reacting with methanesulfonyl chloride, etc.), leaving the 25 alcohol substituents (e.g., ester 26 (X ₂ is converted into halogen or sulfonate) to obtain a halogen or sulfonate leaving group).

26 with an inert solvent (eg, THF / H ₂ O or methylene chloride / H ₂ O or ethyl acetate / H ₂ O or ethanol / H ₂ O or methanol / H ₂ at a temperature of about −25 ° C. to about 100 ° C. The 26 acetal substituents are hydrolyzed to aldehyde 27 by reaction with an acid (eg, trifluoroacetic acid, trifluoromethanesulfonic acid or HCl or acetic acid or sulfuric acid, etc.).

Aldehyde → alcohol reducing agent (eg, hydrogenation) to aldehyde 27 in an inert solvent (eg, THF and / or methylene chloride or ethyl acetate or methyl t-butyl ether or isopropanol) at a temperature of about −25 ° C. to about 100 ° C. Sodium boron or RaNi / H ₂ or borane t-butylamine complex, etc.) is added to give the corresponding alcohol.

The alcohol obtained is about 0.8 to about 3.0 molar equivalents of N— in an inert solvent (such as THF or dioxane or dioxolane or DMF or methylene chloride) at a temperature of about 25 ° C. to about 100 ° C. Reaction with the protected amino acid P ₁ NHCH (R ₁₁ ) COOH or an activated derivative thereof (P ₁ is an N-protecting group, R ₁₁ is isopropyl or isobutyl) provides the diester 28 .

Alternatively, the alcohol is symmetric with a symmetric anhydride derived from P ₁ NHCH (R ₁₁ ) COOH (ie, P ₁ NHCH (R ₁₁ ) C (O) O—C (O) CH (R ₁₁ ) NHP ₁ ). 28 can be obtained by reaction.

28 is reacted with purine 29 in the presence of a base (eg K ₂ CO ₃ etc.) in an inert solvent (eg DMF etc.) to give 30 . Purine 29 is prepared from 6-chloro-2-aminopurine by reaction with R ₉ OH in an inert solvent (eg, toluene or THF) in the presence of a base (eg, NaH or KH). Substitution of substituted purine 30 provides compounds of formula I.

The present invention will now be described by way of example with reference to the following non-limiting examples, comparative examples and the accompanying drawings. In the accompanying drawings:
FIG. 1 is a representation of plasma H2G levels as a function of time in cynomolgus monkeys administered compounds of the invention or other prodrug derivatives of H2G, as further described in Biological Example 3.
FIG. 2 shows the survival as a function of time in herpes simplex-infected mice administered various doses of a compound of the invention or a prior art antiviral agent, as further described in Biological Example 4. It is a thing.

FIG. 1 shows plasma H2G levels as a function of time in cynomolgus monkeys administered a compound of the invention or another prodrug derivative of H2G, as further described in Biological Example 3. FIG. 2 shows the survival of herpes simplex virus-infected mice as a function of time administered various doses of a compound of the invention or a prior art antiviral agent, as further described in Biological Example 4.

(R) -9- [2- (stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine This example illustrates the application of Production Reaction Formula A.
(A) (R) -9- [4- (N-tert-butoxycarbonyl-L-valyloxy) -2- (hydroxymethyl) butyl] guanine Nt-BOC-L-valine (5.58 g, 25. 7 Hg), DMAP (0.314 g, 2.57 mmol) and DCC (6.52 g, 31.6 mmol) were added while heating H2G (5 g, 19.7 mmol) in DMF (300 ml). And cooled to room temperature. The mixture was stirred at room temperature for 24 hours and then filtered. The product was chromatographed on silica gel eluting with CH 2 Cl 2 / MeOH to give 2.4 g of the desired intermediate product.

(B) (R) -9- [4- (N-tert-butoxycarbonyl-L-valyloxy) -2- (stearoyloxymethyl) butyl] guanine Product of step (a) (185 mg, 0.41 mmol) Was dissolved in pyridine (5 ml), the solution was cooled in an ice bath and stearoyl chloride (179 μl, 0.531 mmol) was added. The solution was kept in an ice bath for 2 hours and then at room temperature for 1 hour. This was then evaporated and chromatographed on silica gel. Elution with dichloromethane / methanol gave 143 mg of the desired intermediate product.

(C) (R) -9- [2- (stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine The product of step (b) (138 mg, 0.192 mmol) was cooled in an ice bath. , Trifluoroacetic acid (5 ml) was added. The solution was kept in an ice bath for 45 minutes and then evaporated to give an oil. Water (0.5-1 ml) was added and evaporated twice. The residue was once more dissolved in water (5 ml), filtered and lyophilized to give 148 mg of the desired product as the bistrifluoroacetate salt.

(R) -9- [2- (Myristoyloxymethyl) -4- (L-valyloxy) butyl] guanine In the step (b), myristoyl chloride was used instead of stearoyl chloride, and the title compound was prepared in the same manner as in Example 1. Obtained as the bistrifluoroacetate salt.

(R) -9- [2- (Oleoyloxymethyl) -4- (L-valyloxy) butyl] guanine In the step (b), oleoyl chloride was used instead of stearoyl chloride, and the same procedure as in Example 1 was performed. The compound was obtained as bistrifluoroacetate.

(R) -9- [2- (butyryloxymethyl) -4- (L-valyloxy) butyl] guanine (a) (R) -9- [4- (N-tert-butoxycarbonyl-L-valyloxy) 2- (Butyryloxymethyl) butyl] guanine DCC (110 mg, 0.53 mmol) was dissolved in dichloromethane (10 ml) and butyric acid (82 mg, 0.93 mmol) was added. After 4 hours at room temperature, the mixture was filtered and the filtrate was evaporated. The residue was dissolved in pyridine (5 ml) and (R) -9- [4- (N-tert-butoxycarbonyl-L-valyloxy) -2-hydroxymethyl] butyl] guanine (200 mg, 0.44 mmol) (working) Example 1, step (a)) was added. The mixture was stirred at room temperature for 120 hours. Since the reaction was incomplete by TLC, further anhydride was prepared using the above procedure. The anhydride was added and the mixture was stirred for an additional 20 hours. The reaction mixture was evaporated and chromatographed first on silica gel and then on aluminum oxide, in both cases eluting with dichloromethane / methanol to give 79 mg of intermediate product.

(B) (R) -9- [2- (Butyryloxymethyl) -4- (L-valyloxy) butyl] guanine The intermediate product of step (a) was prepared in the same manner as in step (3) of Example 1. Deprotection yielded 84 mg of the desired product as the bistrifluoroacetate salt.

(R) -9- [2- (Decanoyloxymethyl) -4- (L-valyloxy) butyl] guanine In the step (b), decanoyl chloride was used instead of stearoyl chloride, and the same procedure as in Example 1 was performed. The compound was obtained as bistrifluoroacetate.

(R) -9- [2- (Docosanoyloxymethyl) -4- (L-valyloxy) butyl] guanine A mixture of Nt -BOC-L-valine and DMF as a solvent and dichloromethane in the step (b) The title compound was obtained as the bistrifluoroacetate in the same manner as in Example 1 except that the DMAP / DCC conditions of Example 1 step (a) were used together with docosanoic acid.

(R) -9- [4- (L-Isoleuciloxy) -2- (stearoyloxymethyl) butyl] guanine This example illustrates the application of Production Reaction Formula B.
(A) (R) -9- [2-Hydroxymethyl 4- (t-butyldiphenylsilyloxy) butyl] guanine H2G (2 g, 8 mmol) was coevaporated twice with dry DMF, then dry DMF (120 ml) And suspended in pyridine (1 ml). To the suspension, t-butyldiphenylchlorosilane (2.1 ml, 8.2 mmol) in dichloromethane (20 ml) was added dropwise at 0 ° C. over 30 minutes. When the addition was complete, the reaction mixture became a clear solution. The reaction was continued at 0 ° C. for 2 hours and then kept at 4 ° C. overnight. Methanol (5 ml) was added to the reaction solution. After 20 minutes at room temperature, the reaction mixture was evaporated to a small volume, poured into aqueous sodium bicarbonate and extracted twice with dichloromethane. The organic phase was dried over sodium sulfate and evaporated in vacuo. The product was isolated by silica gel column chromatography using a methanol / dichloromethane system with increasing MeOH concentrations. The product was eluted with 7% MeOH in CH ₂ Cl ₂ to give 1.89 g.

(B) (R) -9- [2- (stearoyloxymethyl) -4- (t-butyldiphenylsilyloxy) butyl] guanine (R) -9- [2-hydroxymethyl 4- (t-butyldiphenylsilyl) Oxy) butyl] guanine (2.31 g, 5 mmol) was co-evaporated twice with dry pyridine and dissolved in pyridine (20 ml). To the solution was slowly added stearoyl chloride (1.86 ml, 5.5 mmol, technical grade) in dichloromethane (2 ml) at -5 ° C. The reaction solution was held at the same temperature for 1 hour and then at 5 ° C. for 2 hours. The reaction was monitored by TLC. Since the reaction was incomplete, stearoyl chloride (0.29 ml) was further added at -5 ° C. After 30 minutes at 5 ° C., methanol (3 ml) was added and the reaction mixture was stirred for 20 minutes. Then, it was poured into an aqueous sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was dried and the product was purified by silica gel column chromatography with MeOH stepping up, eluting with 3.5% MeOH in CH ₂ Cl ₂ (yield 2.7 g).

(C) (R) -9-[(4-hydroxy-2- (stearoyloxymethyl) butyl] guanine (R) -9- [2- (stearoyloxymethyl) -4- (t-butyldiphenylsilyloxy) Butyl] guanine (2.7 g, 3.56 mmol) was dissolved in dry THF (30 ml) and hydrogen fluoride-pyridine (1.5 ml) was added to the solution.The reaction was kept at 4 ° C. overnight and by TLC. The reaction reached about 80% conversion, more HF-pyridine (0.75 ml) was added, and after 4 hours, TLC showed the disappearance of the starting material. Concentrated in vacuo, added more pyridine (5 ml) and evaporated again The product was isolated by silica gel column chromatography (yield 1.26 g).

(D) (R) -9- [4- (N-BOC-L-Isoleuciloxy) -2- (stearoyloxymethyl) butyl] guanine (R) -9- [4-hydroxy-2- (stearoyl) Oxymethyl) butyl] guanine (135 mg, 0.26 mmol) and N-BOC-L-isoleucine (180 mg, 0.78 mmol) were coevaporated twice with dry DMF and dissolved in the same solvent (3.5 ml). did. To the solution was added 1,3-dicyclohexylcarbodiimide (160 mg, 0.78 mmol) and 4-dimethylaminopyridine (4.8 mg, 0.039 mmol). After reacting for 18 hours, the reaction mixture was filtered through Celite and processed as usual. The product was isolated by silica gel column chromatography, eluting with 5% MeOH in CH ₂ Cl ₂ (yield 160 mg).

(E) (R) -9- [4- (L-Isoleuciloxy) -2- (stearoyloxymethyl) butyl] guanine (R) -9- [4- (N-BOC-) in step (d) L-Isoleuyloxy) -2- (stearoyloxymethyl) butyl] guanine (150 mg, 0.205 mmol) was treated with trifluoroacetic acid (3 ml) at 0 ° C. for 20 minutes. The solution was evaporated in vacuo. The residue was coevaporated twice with toluene and kept under vacuum for several hours. The residue was dissolved in MeOH (2 ml) and evaporated to give the trifluoroacetate salt as a glassy product (yield 191 mg).

(R) -9- [2- (decanoyloxymethyl) -4- (L-isoleuyloxy) butyl] guanine In the step (b), decanoyl chloride was used instead of stearoyl chloride, and the same as in Example 7. The title compound was obtained as a bistrifluoroacetyl salt.

(R) -9- [4- (L -Isoleuciloxy ) -2- (myristoyloxymethyl) butyl] guanine In the step (a), N-BOC-L-isoleucine was used instead of N-BOC-valine. The title compound was obtained as a bistrifluoroacetyl salt in the same manner as in Example 1 using myristoyl chloride in step (b).

(R) -9- [2- (4-Acetylbutyryloxymethyl-4- (L-valyloxy) butyl] guanine In the step (b), instead of Nt -Boc-L-valine, the step of Example 1 ( Using the DCC / DMAP conditions of a) with 4-acetylbutyric acid, the title compound was obtained as the bistrifluoroacetate salt as in Example 1.

(R) -9- [2- (Dodecanoyloxymethyl-4- (L-valyloxy) butyl) guanine In the step (b), the title compound was used in the same manner as in Example 1 except that dodecanoyl chloride was used instead of stearoyl chloride. Was obtained as the bistrifluoroacetate salt.

(R) -9- [2-palmitoyloxymethyl-4- (L-valyloxy) butyl] guanine In the step (b), palmitoyl chloride was used instead of stearoyl chloride, and the title compound was converted to bistrifluoromethane in the same manner as in Example 1. Obtained as the acetate salt.

(R) -2-Amino-9- [2-stearoyloxymethyl-4- (L-valyloxy) butyl] purine This example demonstrates the deoxygenation of the group R1.
(A) (R) -2-Amino-9- (2-stearoyloxymethyl-4- (N-tert-butoxycarbonyl-L-valyloxy) butyl) -6-chloropurine:
Example 1 To a solution of (R) -9- (2-stearoyloxymethyl-4- (N-tert-butoxycarbonyl-L-valyloxy) butyl) guanine (646 mg, 0.9 mmol) in acetonitrile in step 2 Tetramethylammonium chloride (427 mg, 2.7 mmol), N, N-diethylaniline (0.716 ml, 4.5 mmol) and phosphorus oxychloride (0.417 ml, 4.5 mmol) were added. The reaction was kept under reflux and progress was monitored by TLC. After 3 hours, the reaction mixture was evaporated in vacuo and the residue was dissolved in dichloromethane and then poured into cold aqueous sodium bicarbonate. The organic phase was evaporated and purified by silica gel column chromatography. Yield: 251 mg.

(B) (R) -2-Amino-9- (2-stearoyloxymethyl-4- (N-tert-butoxycarbonyl-L-valyloxy) butyl) purine:
(R) -2-Amino-9- (2-stearoyloxymethyl-4- (N-tert-butoxycarbonyl-L-valyloxy) butyl) -6 in methanol / ethyl acetate (6 ml, 3: 1 V / V) -To a solution of chloropurine (240 mg, 0.33 mmol) was added ammonium formate (105 mg, 1.65 mmol) and 10% palladium / carbon (15 mg). The reaction was held at reflux for 1 hour and ammonium formate (70 mg) was added again. After an additional hour, TLC showed that the reaction was complete, so the mixture was filtered through Celite and washed thoroughly with ethanol. The filtrate was evaporated and purified on a silica gel column. Yield: 193 mg.

(C) (R) -2-Amino-9- (2-stearoyloxymethyl-4- (L-valyloxy) butyl) purine:
(R) -2-Amino-9- (2-stearoyloxymethyl-4- (N-tert-butoxycarbonyl-L-valyloxy) butyl) purine (180 mg, 0.26 mmol) was added to trifluoroacetic acid at 0 ° C. (5 ml) for 40 minutes. This was then evaporated in vacuo and co-evaporated sequentially with toluene and methanol. The residue was lyophilized overnight to give 195 mg of the desired product.

(R) -9- [4-Hydroxy-2- (stearoyloxymethyl) butyl] guanine other production method (a) Production of ethyl 4,4-diethoxy-2-ethoxycarbonylbutyrate

Potassium tert-butoxide (141.8 g, 1.11 eq) was dissolved in dry DMF (1 L). Diethyl malonate (266 mL, 1.54 equiv) was added over 5 minutes. Bromoacetaldehyde diethyl acetal (172 mL, 1.14 mol) was added over 5 minutes. The mixture was heated to 120 ° C. (internal temperature) and stirred at 120 ° C. for 5 hours. The mixture was allowed to cool to room temperature, poured into water (5 L) and extracted with methyl tert-butyl ether (MTBE, 3 × 600 mL). The organic solution was dried over MgSO ₄ , filtered, concentrated and distilled (0.5 mm, 95-140 ° C.) to give the desired diester (244 g, 78%) as a colorless oil.

(B) Production of 4,4-diethoxy-2- (hydroxymethyl) -butanol

LiBH ₄ (purchased solution, 2M in THF, 22.5 mL) and the product of Example 14 step (a) (5 g in 15. mL THF, 18.1 mmol) were mixed and warmed to 60 ° C., Stir at 60 ° C. for 4 hours. The reaction mixture was allowed to cool to room temperature and the reaction vessel was placed in a cold water bath. Triethanolamine (5.97 mL, 1 eq) was then added at a rate such that the temperature of the reaction mixture was maintained between 20-25 ° C. Saline (17.5 mL) was added at a rate that could control gas evolution and the mixture was stirred at room temperature for 45 minutes. The layers were separated and the organic layer was washed with brine (2 × 15 mL). The combined brine wash was extracted with MTBE (methyl tert-butyl ether, 3 × 20 mL). The combined organic extracts were evaporated and the residue was dissolved in MTBE (50 mL) and washed with brine (25 mL). The brine layer was back extracted with MTBE (3 × 25 mL). The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated to give the desired diol (3.36 g, 15.5 mmol, 97%) as a colorless oil.

(C) Preparation of (2R) -2-acetoxymethyl-4,4-diethoxy-butanol

Place the product of Example 14 step (b) (3.84 g, 20 mmol) in a 10 ml 1 neck round bottom flask followed by vinyl acetate (2.6 g, 30 mmol) and finally lipase PS30 (69 mL, Amano, Lombard, Illinois) was added. The mixture was stirred at ambient temperature for 16 hours. The progress of the reaction was stained with TLC (2/1 hexane-EtOAc; Ce ₂ (SO ₄ ) ₃ and charred on a hot plate; the diol r.f. was 0.1 and the monoacetate was 0. .3, bisacetate is 0.75). The reaction mixture was diluted with CH ₂ Cl ₂ and filtered through a 5 micron filter. The filter was further washed with CH ₂ Cl ₂ . The filtrate was then concentrated in vacuo to give the desired product.

(D) Production of (2S) -2-acetoxymethyl-4,4-diethoxybutyltoluenesulfonate

Crude product of Example 14 step (c) (4.62 g, 19 mmol) under dry N _{2 in} a 100 ml 1 neck round bottom flask equipped with a magnetic stir bar and septum, dry CH ₂ Cl ₂ (20 mL) and _Et 3 N _(5.62mL, 40 mmol) was added. To this solution was added tosyl chloride (4.76 g, 25 mmol). The resulting mixture was stirred at ambient temperature for 4 hours. H ₂ O (0.27 g, 15 mmol) was added and stirred vigorously for 4 hours. The reaction mixture was diluted with 80 mL EtOAc and 50 mL H ₂ O and the aqueous layer was separated. To the organic layer was added 75 mL of 5% KH ₂ PO ₄ aqueous solution. After mixing and separating the layers, the aqueous layer was removed. The organic layer was washed with 50 mL saturated NaHCO ₃ solution, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to a desired weight of 7.40 g.

の製造 (E)

Manufacturing of

The product of Example 14 step (d) (3.88 g, 10 mmol), anhydrous DMF (20 mL), 2-amino-4-chloro-purine (2.125 g, 12) in a 50 mL 1 neck round bottom flask. 0.5 mmol) and K ₂ CO ₃ (4.83 g) were added. The resulting suspension was stirred at 40 ° C. for 20 hours under N ₂ blanket. The mixture was concentrated on a rotary evaporator to remove most of the DMF. The residue was diluted with EtOAc (50 mL) and H ₂ O (50 mL). The reaction mixture was transferred to a separatory funnel, shaken and the aqueous layer was separated. The aqueous layer was extracted with EtOAc (25 mL). The organic layers were combined and washed with 5% KH ₂ PO ₄ (75 mL). The organic layer was separated, washed with H ₂ O (75 mL), brine (75 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give 3.95 g of crude product. The crude product was slurried with 40 mL methyl-t-butyl ether. The mixture was stirred overnight at 4 ° C. and the mixture was filtered. The filtrate was concentrated to give 3.35 g of product as an oil (containing 2.6 g of the desired product based on HPLC analysis).

の製造 (F)

Manufacturing of

Benzyl alcohol (136 mL) was added to a 500 mL 1 neck round bottom flask, cooled to 0 ° C., and then KO-t-Bu (36 g, 321 mmol) was added in small portions. The temperature was warmed to 40 ° C. and the mixture was stirred for 20 minutes. To this mixture was added the crude product of Example 14 step (e) (24.7 g, 64.2 mmol) dissolved in 25 mL anhydrous THF and benzyl alcohol (30 mL) at 0 ° C. The temperature was slowly warmed to 8 ° C over 2 hours. The reaction mixture was poured into 500 mL ice and extracted with 500 mL MTBE. The organic layer was washed with 250 mL of brine, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give 193 g of the desired product in benzyl alcohol. HPLC analysis indicated that this solution contained 25.96 g of the desired product.

の製造 (G)

Manufacturing of

The crude product of Example 14 step (f) (1.30 g, 3.13 mmol of Example 14 step (f) dissolved in absolute EtOH (20 mL) in a 100 mL 1 neck round bottom flask. 9.65 g) of benzyl alcohol solution containing the product was added. To this was added 0.45 g of 10% Pd / C slurried in 5 mL of absolute EtOH. The reaction flask was evacuated, it was added 3 times with _{H 2} balloon for _{H 2.} The reaction flask was pressurized with 1 atm H ₂ and the reaction mixture was stirred overnight. The reaction mixture was filtered through a pad of diatomaceous earth to remove Pd / C. Volatile components were removed under vacuum. The residue was mixed with 25 mL of isopropyl acetate and then concentrated in vacuo. The residue was diluted with EtOAc (10 mL), seeded with the desired product, heated to reflux, then CH ₃ CN (2 mL) and MTBE (35 ml) were added. The mixture was stirred for 30 minutes. The precipitate was filtered and the desired product was dried to a constant weight of 600 mg.

の製造 (H)

Manufacturing of

In a 25 mL 1-neck round bottom flask, add the product of Example 14 step (g) (0.650 g, 2.0 mmol), pyridine (4 mL) and CH ₂ Cl ₂ (2 mL), DMAP (10 mg). added. The mixture was cooled to −5 ° C. and stearoyl chloride (790 mg, 2.6 mmol) dissolved in CH ₂ Cl ₂ (0.5 mL) was added over 5 minutes. The resulting mixture was stirred at −5 ° C. for 16 hours. Anhydrous EtOH (0.138 g, 3.0 mmol) was added and the mixture was stirred for an additional hour. The reaction mixture was concentrated in vacuo. Toluene (30 mL) was added to the residue and then the mixture was concentrated in vacuo. Toluene (30 mL) was again added to the residue and then the mixture was concentrated in vacuo. To the residue was added 1% KH ₂ PO ₄ (25 mL) and the mixture was extracted with CH ₂ Cl ₂ (60 mL). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to a constant weight of 1.65 g. The crude product was chromatographed on 40 g SiO ₂ and eluted with 95/5 CH ₂ Cl ₂ -EtOH to give 367 mg of the desired product.

の製造 (I)

Manufacturing of

The product of Example 14 step (h) (0.234 g, 0.394 mmol) dissolved in THF (1.7 mL) was added to a 25 mL 1 neck round bottom flask. To this solution was added trifluoromethanesulfonic acid (0.108 g) in H ₂ O (180 mg). The mixture was stirred overnight at room temperature. To the reaction mixture was added saturated NaHCO ₃ solution (10 mL), THF (5 mL), CH ₂ Cl ₂ (2 mL) and NaBH ₄ (0.10 g). The mixture was stirred for 30 minutes. To the reaction mixture was added a 5% solution of KH ₂ PO ₄ (30 mL). This mixture was extracted with 2 × 15 mL of CH ₂ Cl ₂ . The organic layers were combined, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to a constant weight of 207 mg. This material was recrystallized from EtOAc (8 mL) and CH ₃ CN (0.5 mL) to give 173 mg of the desired product.

Other production methods for (R) -9- [4- (N-tert-butyloxycarbonyl-L-valyloxy) -2- (stearoyloxymethyl) butyl] guanine (R) -9- [2- (stearoyloxy) Methyl) -4- (t-butyldiphenylsilyloxy) butyl] guanine (45 g) and THF (950 ml) were combined in a 2 L flask. Boc-L-valine (3.22 g, 0.25 equiv) was then added, followed by tetrabutylammonium fluoride (1M in THF, 89.05 mL) over 10 minutes. The clear reaction mixture was stirred at room temperature for 2 hours 50 minutes and the progress of the reaction was monitored by TLC (90/10 CH ₂ Cl ₂ / MeOH).

To the reaction mixture was added Boc-L-valine (35.43 g, 2.75 eq), DCC (36.67 g, 2.75 eq) and dimethylaminopyridine (1.1 g, 0.15 eq) in THF (25 ml). Was added. The reaction mixture was stirred at room temperature for 24 hours. DCU was filtered off and washed with CH ₂ Cl ₂ . The filtrate was concentrated and the residue was taken up in 2 liters of CH ₂ Cl ₂ and washed with 2 L of 1/2 saturated sodium bicarbonate solution and brine solution. Drying and evaporation gave about 100 g of crude product. This material was purified by silica chromatography (6000 ml of silica) using 3% MeOH / CH ₂ Cl _{2 to} 5% MeOH / CH ₂ Cl ₂ to give 38.22 mg of the desired product.

(R) -9- [2- (stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine other production method (a) (R) -9- [2-hydroxymethyl) -4- (t -Butyldiphenylsilyloxy) butyl] guanine H2G (45.0 g, 1.78 mol) and N, N dimethylformamide (6.4 kg) were placed in a Bucchi evaporator and the mixture was warmed to dissolve the solid. The solution was concentrated to dryness under vacuum below 90 ° C. The resulting powder was transferred to a 22 liter flask equipped with a stirrer, addition funnel and temperature probe. N, N-dimethylformamide (1.7 kg) was added followed by pyridine (3.53 kg). The resulting suspension was cooled to −10 ° C. under nitrogen and stirred at −5 ± 5 ° C. while adding t-butylchlorodiphenylsilane (684 g, 2.49 mol) dropwise. The resulting mixture was reacted until complete (TLC (10: 1 methylene chloride / methanol) and HPLC (4.6 × 250 mm Zorbax RxC8 (5 microns); 1.5 ml / min 60:40 acetonitrile- Monitored by NH ₄ OAC aqueous solution (0.05 M); UV detection at 254 nm). Water (16 kg) was added and the mixture was stirred for 30 minutes to precipitate the product, then the mixture was cooled to 0 ° C. for 30 minutes. The solid was isolated by filtration and the product cake was washed with cold water and sucked dry with air to give the crude product as an off-white solid. The crude solid was taken up in pyridine (3 kg) and concentrated under vacuum at 60 ° C. to remove water. The dry solid residue was slurried with methanol (10 kg) at 60 ° C. for 1-2 hours and filtered while hot. The filtrate was concentrated in vacuo and the solid residue was refluxed with isopropyl acetate (7 kg) for 30 minutes. The mixture was cooled to 20 ° C. and filtered. The filter cake was dried in vacuo at 50 ° C. to give the title compound as a white solid (555 g).

(B) (R) -9- [2- (stearoyloxymethyl) -4- (t-butyldiphenylsilyloxy) butyl] guanine The product of Example 16 step (a) (555 g, 1.113 mol) was obtained. Placed in a 50 liter butch evaporator. Pyridine (2.7 kg) was added dropwise to dissolve the solid and the mixture was evaporated to dryness at 60 ° C. under vacuum. The residue was taken up in fresh pyridine (2.7 kg) and transferred to a 22 liter flask equipped with a stirrer, addition funnel and temperature probe. The solution was cooled to −5 ° C. under nitrogen. A solution of stearoyl chloride (440 g, 1.45 mol) in methylene chloride (1.5 kg) was added so that the temperature was below 0 ° C. 4- (N, N-dimethylamino) pyridine (15 g, 0.12 mol) was added and until conversion was complete (TLC (10: 1 methylene chloride / methanol) and HPLC (4.6 × 250 mm Zorbax RxC8 (5 (Micron); monitored by 60:40 acetonitrile-NH ₄ OAC aqueous solution (0.05 M); UV detection at 254 nm) at 1.5 ml / min) The mixture was stirred at -5 to 0 ° C. for 2 to 4 hours. At the end of the reaction, acetonitrile (8.7 kg) was added and the mixture was stirred for more than 15 minutes to precipitate the product. The slurry was cooled at 0 ° C. for 2 hours, the solid was isolated by filtration, and the filter cake was washed with acetonitrile (2 kg). The desired product was obtained as a white solid (775 g).

(C) (R) -9- [4-Hydroxy-2- (stearoyloxymethyl) butyl] guanine A solution of the product of Example 16 step (b) (765 g, 0.29 mol) in tetrahydrofuran (10 kg). Was prepared in a reactor. A solution of tetra (n-butyl) ammonium fluoride in tetrahydrofuran (1.7 kg of 1M solution, 1.7 mol) was added and the resulting clear solution was stirred at 20 ± 5 ° C. for 4 hours. Water (32 kg) was added and the resulting slurry was stirred for 1 hour and then cooled to 0 ° C. for 30 minutes. The precipitate was isolated by filtration and the filter cake was washed sequentially with water (10 kg) and acetonitrile (5 kg). After drying at 25 ° C. under vacuum, 702 g of crude product was obtained. The crude product was dissolved in refluxing THF (4.2 kg) and water (160 g), then cooled to 40 ° C. and treated with methylene chloride (14.5 kg). The mixture was cooled to 25 ± 5 ° C. for 1 hour and then cooled to 5 ± 5 ° C. for 1 hour to complete the precipitation. A slightly off-white powder was isolated by filtration and dried under vacuum at 40 ° C. to give the desired product (416 g).

(D) (R) -9- [4- (N-Cbz-L-valyloxy) -2- (stearoyloxymethyl) butyl] guanine N-Cbz-L-valine (169 g, 0. 1 in THF (750 ml)). 67 mol) was prepared in a 2 liter flask equipped with a mechanical stirrer, thermometer and addition funnel. A solution of dicyclohexylcarbodiimide (69.3 g, 0.34 mol) in THF (250 ml) was added over 5 minutes and the resulting slurry was stirred at 20 ± 5 ° C. for 2 hours. The slurry was filtered and the filter cake was washed with THF (300 ml). The filtrate and washings were placed in a 3 liter flask equipped with a stirrer and a thermometer. The product of Example 16 step (c) (116 g, 0.22 mol) was added as a solid and rinsed with THF (250 ml). 4- (N, N-dimethylamino) pyridine (2.73 g, 0.022 mol) was added and the white slurry was stirred at 20 ± 5 ° C. All solids dissolved within 15 minutes and the reaction was completed within 1 hour (determined by HPLC: 4.6 × 250 mm Zorbax RxC8 column; 85 ml of 1 ml / min 85:15 acetonitrile-0.2% HClO ₄ aqueous solution; 254 nm UV detection; starting material elutes at 4.1 minutes and product elutes at 5.9 minutes). Water (5 ml) was added to quench the reaction and the solution was concentrated in vacuo to give a pale yellow semi-solid. This was taken up in methanol (1.5 liters) and heated to reflux for 30 minutes. The solution was cooled to 25 ° C. and the precipitate was removed by filtration. The filtrate was concentrated in vacuo to give a viscous light yellow oil. Acetonitrile (1 L) was added and the resulting white suspension was stirred at 20 ± 5 ° C. for 90 minutes. The crude solid product was isolated by filtration, washed with acetonitrile (2 × 100 ml) and air dried overnight to give the desired product as a waxy sticky solid (122 g). This was further crystallized from ethyl acetate (500 ml) and purified by vacuum drying at 30 ° C. to give the desired product as a white waxy solid (104 g).

(E) (R) -9- [4- (L-valyloxy) -2- (stearoyloxymethyl) butyl] guanine Production of Example 16 step (d) in warm (40 ° C.) ethanol (2.3 L) A solution of product (77 g) was placed in a hydrogenation reactor with 5% Pd-C (15.4 g). The mixture was stirred vigorously at 40 ° C. under 40 psi hydrogen pressure for 4 hours, evacuated and hydrogenated for an additional 4-10 hours. The catalyst was removed by filtration and the filtrate was concentrated in vacuo to give a white solid. This was stirred with ethanol (385 ml) at 25 ° C. for 1 hour, then cooled to 0 ° C. and filtered. The filter cake was air dried and then dried under vacuum at 35 ° C. to give the title compound as a white powder (46 g).

(R) -9- [2- (L-valyloxy) -4- (stearoyloxymethyl) butyl] guanine (a) (R) -9- [2-hydroxymethyl-4- (stearoyloxy) butyl] guanine H2G (506 mg; 2.0 mmol) was dissolved in dry N, N-dimethylformamide (40 ml) with pyridine (400 mg; 5.06 mmol) and 4-dimethylaminopyridine (60 mg; 0.49 mmol). Stearoyl chloride (1500 mg; 4.95 mmol) was added and the mixture was kept at room temperature overnight. Most of the solvent was evaporated in vacuo, the residue was stirred with 70 ml of ethyl acetate and 70 ml of water, the solid was filtered off, washed with ethyl acetate and water and dried to give 680 mg of crude product. Column chromatography on silica gel (chloroform: methanol 15: 1) gave the pure title compound as a white solid.

(B) (R) -9- [2- (N-Boc-L-valyloxymethyl) -4- (stearoyloxy) butyl] guanine N-Boc-L-valine (528 mg; 2 in dichloromethane (20 ml)) .1 mmol) and N, N′-dicyclohexylcarbodiimide (250 mg; 1.21 mg) are stirred overnight at room temperature, dicyclohexylurea is filtered off and extracted with a little dichloromethane, and the filtrate is evaporated in vacuo to a small volume. It was. (R) -9- [2-hydroxymethyl-4- (stearoyloxy) butyl] guanine (340 mg; 0.654 mmol), 4-dimethylaminopyridine (25 mg; 0.205 mmol) and dry N, N-dimethyl Formamide (15 ml) was added and the mixture was stirred at 50 ° C. under N 2 for 4 hours. The solvent was evaporated in vacuo to a small volume. Column chromatography on silica gel followed by aluminum oxide (ethyl acetate: methanol: water 15: 2: 1 as eluent) gave 185 mg (39%) of the pure title compound as a white solid.

(C) (R) -9- [2- (L-valyloxymethyl) -4- (stearoyloxy) butyl] guanine (R) -9- [2- (N-Boc-L-valyloxymethyl)- To 4- (stearoyloxy) butyl] guanine (180 mg; 0.25 mmol) is added cold trifluoroacetic acid (2.0 g), the solution is kept at room temperature for 1 hour, evaporated to a small volume, white amorphous It was repeatedly lyophilized with dioxane until a powder was obtained. The yield of the title compound obtained as the trifluoroacetate salt was quantitative.

Other Preparation of (R) -9- [2-Hydroxymethyl-4- (stearoyloxy) butyl] guanine H2G (7.60 g, 30 mmol) was dissolved by heating in dry DMF (200 ml). The solution is filtered to remove solid impurities, cooled to 20 ° C. (H2G crystallizes), pyridine (9.0 g, 114 mmol), 4-dimethylaminopyridine (0.46 g, 3.75 mmol). Stirred at this temperature while addition and then stearoyl chloride (20.0 g, 66 mmol) was added slowly. Stirring was continued overnight at room temperature. Then most of the solvent was removed in vacuo, the residue was stirred with 200 ml ethyl acetate and 200 ml water, the solid was filtered off, washed with ethyl acetate and water and dried to give the crude product. As an alternative to recrystallization, the crude product was heated with 100 ml ethyl acetate: methanol: water (15: 2: 1) for a while until it boiled, the suspension was slowly cooled to 30 ° C. and filtered. Most of the 2 "isomer in solution (2" isomer crystallizes at lower temperatures) was obtained. After repeating the extraction procedure one more time and vacuum drying, 6.57 g (42%) of the product containing almost no isomer was obtained.

Preparation of crystalline (R) -9- [2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine The product of Example 16 step (c) (20.07 g, 32.5 mmol) was anhydrous. Heat dissolved in ethanol (400 ml), filtered and further diluted with ethanol (117.5 ml). To this solution was added water (HPLC grade, 103.5 ml) and the mixture was cooled to 35-40 ° C. After cooling the mixture, water (HPLC grade, 931.5 ml) was added at a constant rate over 16 hours with good stirring. After all the water was added, stirring was continued for 4 hours at room temperature. The resulting precipitate was filtered through paper and dried under vacuum at room temperature to give the title compound as a white free flowing crystalline powder (19.43 g, 97%) (mp 169- 170 ° C).

9-R- (4-hydroxy-2- (L-valyloxymethyl) butyl) guanine (a) 9-R- (4- (tert-butyldiphenylsilyloxy) -2- (hydroxy) in DMF (30 ml) To a solution of (methyl) butyl) guanine (695 mg, 1.5 mmol) was added N-Boc-L-valine (488 mg, 2.25 mmol), 4-dimethylaminopyridine (30 mg, 0.25 mmol) and DCC (556 mg). 2.7 mmol) was added. After 16 hours, N-Boc-L-valine (244 mg) and DCC (278 mg) were added again to the reaction solution, and the mixture was further maintained for 5 hours. The reaction mixture was filtered through celite, poured into an aqueous sodium bicarbonate solution, and then extracted with dichloromethane. The organic phase was evaporated and purified by silica gel column chromatography to give 950 mg of N-protected monoaminoacyl derivative.

(B) The above intermediate (520 mg, 0.78 mmol) was dissolved in THF (15 ml). To the solution was added hydrogen fluoride in pyridine (70% / 30%, 0.34 ml). After 2 days, the solution was evaporated and coevaporated with toluene. Purification by silica gel column chromatography gave 311 mg of protected monoaminoacyl compound.

(C) The product of step (b) (95 mg, 0.21 mmol) was treated with a mixture of trifluoroacetic acid (4 ml) and dichloromethane (6 ml) for 1 hour. The solution was evaporated and lyophilized to give 125 mg of deprotected monoaminoacyl product.

(R) -9- (2-hydroxymethyl-4- (L-isoleuyloxy) butyl) guanine (a) (R) -9- (2-hydroxymethyl-4-hydroxybutyl in DMF (250 ml) ) To a solution of guanine (2.53 g, 10 mmol), N-Boc-L-isoleucine (2.77 g, 12 mmol), 4-dimethylaminopyridine (61 mg, 0.6 mmol) and DCC (3.7 g, 18 mmol) was added. After 16 hours of reaction at 0 ° C., N-Boc-L-isoleucine (1.3 g) and DCC (1.8 g) were added again and the reaction was kept at room temperature overnight. The reaction mixture was filtered through Celite and the filtrate was evaporated and purified by silica gel column chromatography to give 1.25 g of N-protected monoaminoacyl intermediate.

(B) The intermediate of step (a) (100 mg, 0.21 mmol) was treated with trifluoroacetic acid (3 ml) at 0 ° C. for 30 minutes. The solution was evaporated and lyophilized to give the title deprotected monoaminoacyl product in quantitative yield.

(R) -9- [2-Hydroxymethyl-4- (L-valyloxy) butyl] guanine The product of Example 1 step (a) was deprotected with trifluoroacetic acid as in Example 1 step (c). did.

(R) -9- [2- (L-valyloxymethyl) -4- (L-valyloxy) butyl] guanine (a) (R) -9- [4- (N-Boc-L-valyloxy) -2 -(N-Boc-L-valyloxymethyl) butyl] guanine Example 1 step (a) with 2.7, 0.28 and 3.2 equivalents of N-Boc-valine, DMAP and DCC, respectively To give the title compound.

(B) (R) -9- [4- (L-valyloxy) -2- (L-valyloxymethyl) butyl] guanine The intermediate of Example 20 step (a) is the same as Example 1 step (c). To give the title compound as the trifluoroacetate salt.

(R) -9- [4-Hydroxy-2- (stearoyloxymethyl) butyl] guanine The title compound is prepared according to steps (a) to (c) of Example 7.

(R) -9- [2-Hydroxymethyl-4- (stearoyloxy) butyl] guanine The title compound is prepared by the procedure of Example 17 step (a).

(R) -9- [2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine other process (a) (R) -9- [4-N-benzyloxycarbonyl-L-valyloxy ) 2- (Hydroxymethyl) butyl] guanine Dry H2G (252 mg, 1 mmol), 4-dimethylaminopyridine (122 mg, 1 mmol) and N-Cbz-L-valine p-nitrophenyl ester (408 mg, 1.1 mmol) ) Was dissolved in dry dimethylformamide (16 ml). After stirring at 23 ° C. for 30 hours, the organic solvent was removed and the residue was carefully chromatographed (silica, 2% -7% methanol / methylene chloride) to give the desired product as a white solid (151 mg, 31%). .

(B) (R) -9- [4-N-Benzyloxycarbonyl-L-valyloxy) 2- (stearoyloxymethyl) butyl] guanine stearoyl chloride (394 mg, 1.3 mmol) in dry methylene chloride (2 ml) Was slowly added dropwise at −5 ° C. under nitrogen to a solution of the product of step (a) (243 mg, 1 mmol) and 4-dimethylaminopyridine (20 mg) in dry pyridine (5 ml). The reaction mixture was stirred at this temperature for 12 hours. Methanol (5 ml) was added and the reaction was stirred for 1 hour. After removal of the solvent, the residue was triturated with acetonitrile and chromatographed (silica, 0-5% methanol / methylene chloride) to give the desired product (542 mg, 72%).

(C) (R) -9- [2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine The product of step (b) (490 mg, 1 mmol) was dissolved in methanol (30 ml), 5% Pd / C (100 mg) was added. A balloon filled with hydrogen was placed on top of the reaction vessel. After 6 hours at 23 ° C., TLC showed the absence of starting material. The reaction mixture was filtered through a 0.45 micron nylon membrane to remove the catalyst and the solvent was removed to give the desired product as a white solid (350 mg, 99%). This was identical to that obtained in Example 16 (spectrum and analytical data).

Other production methods of (R) -9- (4-hydroxy-2- (L -valyloxymethyl) butyl) guanine Example 23 Step (b) (R) -9- (4- (L-valyloxy) 2- (L-valyloxymethyl) butyl) guanine (100 mg, 0.126 mmol) was dissolved in 0.1 N aqueous NaOH (6.3 ml, 0.63 mmol) at room temperature. Periodic aliquots were taken and neutralized with 0.5N trifluoroacetic acid. An aliquot was evaporated and analyzed by HPLC to monitor the progress of the reaction. After 4 hours, 0.5N trifluoroacetic acid solution (1.26 ml, 0.63 mmol) was added to the solution and the reaction mixture was evaporated. The desired product was purified by HPLC (YMC, 50 × 4.6 mm, gradient 0.1% TFA + 0-50% 0.1% TFA in acetonitrile, 20 min UV detection at 254 nm) (yield: 13.6%).

(R) -9- (2-Hydroxymethyl-4- (L-valyloxy) butyl) guanine Other Preparation Method The reaction solution of Example 27 was separated by HPLC to give the title compound in a yield of 29.2%. It was.

(R) -9- [2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine monohydrochloride The product of Example 16 step (d) (360 mg, 0.479 mmol) was added to methanol (10 ml). And dissolved in a mixture of ethyl acetate (10 ml). To the solution was added 10% Pd / C (100 mg) and 1N HCl (520 μl). The reaction mixture was stirred at room temperature for 2 hours at 1 atm H2. The reaction mixture was filtered and the solvent was evaporated from the filtrate to give the desired product as a crystalline solid (300 mg).

Other production methods of (R) -9-[(2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine (a) (R) -2-amino-6-chloro-9- [4, Preparation of 4-diethoxy-2- (hydroxymethyl) butyl] purine The product of Example 14 step (e) (200 g) was dissolved in methanol (670 mL) and 20% aqueous K ₂ CO ₃ was added. The mixture was stirred at 25 ± 5 ° C. for 30 minutes. The reaction mixture was then cooled to 0-5 ° C. for about 20 minutes and a precipitate formed. Water (500 mL) was added and the slurry was mixed at 5 ± 5 ° C. for 15 minutes. The resulting solid was isolated by filtration and the filter cake was washed with water (100 mL) and dried under vacuum at 20 ° C. to give the desired product as a pale yellow powder (81 g).

(B) Preparation of (R) -9- [4,4-diethoxy-2- (hydroxymethyl) butyl] guanine Example 30 Step (a) product (22.5 kg, 65.4 mol) was added to a KOH aqueous solution. (Prepared by dissolving 12.9 kg of KOH in 225 kg of water). The mixture was refluxed for 16 hours. The reaction was cooled to approximately room temperature and filtered into a larger reactor equipped with a pH electrode standardized to pH 7-10. The filtered solution is cooled to 5 ° C. and the product is diluted with dilute acetic acid solution (glacial acetic acid (12.6 kg, 210 mol) until the pH is between 7.5 and 9.0 (target 8.5). It was precipitated by mixing slowly with 75 kg of water and preparing the mixture by cooling to 5 ° C. The resulting slurry was immediately filtered and the filter cake was refilled in the reactor. The reactor was charged with 225 kg of distilled water. The mixture was heated to below 50 ° C. for 30 minutes, then cooled to 15 ± 10 ° C. and stirred for 30 minutes. The resulting precipitate was filtered by vacuum filtration, rinsed with 50 kg distilled water, and vacuum dried in an oven at less than 45 ° C. for at least 8 hours to give the desired product as a tan solid.

(C) Preparation of stearoyl-pivaloyl mixed anhydride Triethylamine (8.2 kg, 81.0 mol) was added to stearic acid (22.4 kg, 78.7 mol) in toluene (156.4 kg). The internal temperature of the resulting slurry was lowered to −5 ° C., and then pivaloyl chloride (9.52 kg 79.0 mol) was slowly added while maintaining the internal temperature not exceeding 5 ° C. The slurry was stirred at 5 ° C. for 2 hours, then warmed to 20 ° C. and stirred for 4 hours. The triethylammonium hydrochloride precipitate was filtered and washed with 36.6 kg, 35.5 kg and 37.9 kg toluene. The filtrate was concentrated at an internal temperature not exceeding 60 ° C., 61.1 kg heptane was added, and then the slurry was cooled to −15 to −10 ° C. After stirring for 4 hours, the resulting solid was collected by vacuum filtration, blown dry with nitrogen gas for 1 hour, and dried in a vacuum oven at room temperature to give the desired product as white crystals (18.9 g). It was. The mother liquor was concentrated in vacuo and 41.1 kg of heptane added to give an additional 2.7 kg of the desired product. The resulting slurry was cooled to −15 to −10 ° C. for 4 hours, filtered, blown dry with nitrogen gas for 1 hour, and the product was dried in a vacuum oven at room temperature.

(D) Preparation of (R) -9- [4,4-diethoxy-2- (stearoyloxymethyl) butyl] guanine The product of Example 30 step (b) (3.9 kg, 11.9 mol), carried out The product of Example 30 step (c) (5.2 kg, 13.6 mol) and 4-dimethylaminopyridine (300 g, 2.4 mol) were mixed in THF (103.3 kg) at room temperature. After mixing for 16 hours, water (3 kg) was added. After mixing for 45 minutes, the solution was distilled at an internal temperature not exceeding 45 ° C. Ethyl acetate (62.9 kg) was added and the solution redistilled at an internal temperature not exceeding 45 ° C. Acetone (56 kg) was then added and the slurry was heated to reflux (56 ° C.) for 15 minutes. The resulting clear solution was cooled to room temperature (not exceeding 15 ° C./hour). After 4 hours at room temperature, the resulting precipitate was filtered and rinsed with acetone (17 kg).

  The mother liquor was concentrated under vacuum at a temperature not exceeding 45 ° C. Ethyl acetate (260 kg) and water (72.1 kg) were added. The bilayer mixture was stirred and then allowed to settle. The organic phase was separated and distilled. Ethyl acetate (200 kg) was added and the solution redistilled. Acetone (101 kg) was added and the solution was heated to reflux (56 ° C.) for 15 minutes, then the solution was cooled to room temperature (do not exceed 15 ° C./hour) and the precipitate was filtered. The product is washed with acetone (19 kg, 15 kg and 15 kg), blown dry with nitrogen gas for 1 hour and then vacuum dried at a temperature not exceeding 40 ° C. for about 6 hours to give the desired product (3.1 kg). Got.

(E) Preparation of (R) -9- [4-hydroxy-2- (stearoyloxymethyl) butyl] guanine The product of Example 30 step (d) (3.0 kg) was dissolved in THF (46 L) at 20 ° C. To make a slurry. A solution of trifluoromethanesulfonic acid (2.25 kg) in water (2.25 kg) (prepared by slowly adding the acid to cold water) was added and the reaction mixture was stirred at 22 ° C. for 2 hours. The reaction mixture was cooled to 15 ° C. and quenched with a solution of NaHCO ₃ (1.5 kg) in water (5.3 kg).

  Borane t-butylamine complex (powder, 340 g) was added in four portions, then the reaction temperature was raised to 35 ° C. and stirred for 12 hours. The reaction mixture was added to a solution of concentrated HCl (37% aqueous solution, 320 g) in tap water (115 kg) at 5 ° C. The mixture was stirred for 30 minutes and the resulting precipitate was filtered and washed with acetonitrile (15 kg). The solid was reprecipitated once or twice from acetone (35 kg). The product was dissolved in THF (24 kg) at 65 ° C., water (1.3 kg) was added, cooled to 30 ° C., and then methylene chloride (105 kg) was added for final precipitation. The resulting slurry was cooled to 10 ° C. and the precipitate was filtered to give the desired product.

(F) Preparation of (R) -9- [4- (N-benzyloxycarbonyl-L-valyloxy) -2- (stearoyloxymethyl) butyl] guanine N-carbobenzyloxy-L- in THF (20 L) To a reactor containing a mixture of valine (3630 g, 14.5 mol) was added a solution of dicyclohexylcarbodiimide (1500 g, 7.27 mol) in THF (7 L). The resulting mixture was stirred at 20 ± 5 ° C. for 1-2 hours. The product of Example 30 step (e) (2500 g, 4.81 mol) and 4-dimethylaminopyridine (59 g, 0.48 mol) were placed in a second reactor. To this second reactor, the THF mixture from the first reactor was filtered and then rinsed with THF (15 L). The resulting mixture was stirred at 20 ± 5 ° C. for 1-3 hours. Water (600 mL) was added and the solution was concentrated in vacuo at a temperature not exceeding 45 ° C. The residual oil was taken up in ethyl acetate (14 L) and filtered. The filtrate was washed sequentially with 10% aqueous sodium bicarbonate (2 × 14 L) and 10% brine (14 L). The organic phase was concentrated in vacuo and the residue was dissolved in methanol (10 kg) at 50-60 ° C. This warm solution was slowly added to a mixture of acetonitrile (30 kg) and water (13 kg) at ambient temperature. The mixture was stirred at 15 ° C. for 1 hour, then filtered to isolate the crude product, which was dried in vacuo at 40 ° C. to give the desired product as a white solid (3.9 kg).

(G) Preparation of (R) -9-[(2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine 10% Pd-C (400 g) in a hydrogenation reactor and Example 30 step (f) Product (2.4 kg) was added. Absolute ethanol (52 L) was added and the mixture was warmed to 40 ° C. and hydrogenated at 30-40 psi for 3-5 hours. After the reaction was complete, the catalyst was filtered off through diatomaceous earth and the filter cake was thoroughly rinsed with ethanol (30 L). The combined filtrate was concentrated in vacuo at a temperature not exceeding 60 ° C. to give a white residue. This was dissolved in isopropanol (15 L) and isopropyl acetate (60 L) under reflux and then cooled to room temperature over 4 hours. After cooling at 15 ± 10 ° C. for 3 hours, the precipitate was isolated by filtration, washed with isopropyl acetate (6 L) and dried in vacuo at 40 ° C. to give the desired product as a white powder (864 g).

(R) -9-[(2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine other production method (a) of (2R) -4,4-diethoxy-2-stearoyloxymethylbutanol Manufacturing

Vinyl stearate (17.76 g, 0.057 mol) was placed in a 100 mL round bottom flask equipped with a magnetic stir bar. The flask was immersed in a 35 ° C. oil bath with stirring. Example 14 The product of step (b) (10.0 g, 0.052 mol) and lipase Amano PS-30 (0.20 g) were added and stirred at 35 ° C. for 4 hours. The reaction was diluted with hexane (260 mL) and MTBE (115 mL) and filtered through celite. The filtrate was washed twice with water (100 mL), dried over Na ₂ SO ₄ and concentrated to give the desired product (26.21 g) as a clear oil (forms a wet solid on standing at room temperature). It was.

(B) Production of (2S) -4,4-diethoxy-2-stearoyloxymethylbutyltoluenesulfonate

Example 31 The product of step (a) (26.21 g, 0.057 mol) was dissolved in methylene chloride (75 mL) and a 250 mL volume equipped with a magnetic stir bar, condenser, N ₂ inlet and temperature probe. Placed in a three-necked flask. Triethylamine (14.4 g) was added followed by p-toluenesulfonyl chloride (16.3 g). N ₂ was passed through the flask and heated to reflux (46 ° C.). The reaction was stirred at reflux for 6 hours. The reaction was cooled to room temperature. Water (10 mL) was added and the reaction was stirred vigorously for 16 hours. The reaction mixture was poured into a 1 L separatory funnel containing ethyl acetate (350 mL) and water (350 mL). The organic layer was separated and washed with 7% (w / w) aqueous sodium bicarbonate (100 mL). The organic layer was then washed with 23% (w / w) aqueous sodium chloride solution (100 mL). The organic layer was dried over Na ₂ SO ₄ and filtered. The solution was concentrated to give the desired product (29.4 g) as an oil (forms a wet solid upon standing at room temperature).

(C) Production of (3S) -3-stearoyloxymethyl-4-toluenesulfonyloxybutyraldehyde

Example 31 Step (b) product (29.38 g; assay performed at 23.12 g, 0.037 mol) dissolved in THF (90 mL), 250 mL volume with magnetic stir bar and temperature probe. Place in a round bottom flask. Water (38 mL) was added and cooled to 10 ° C. Trifluoroacetic acid (55 mL) was poured and the mixture was stirred for 25 minutes. The reaction mixture was poured into a 2 L separatory funnel containing 20% (w / w) K ₂ CO ₃ solution (690 g), ice (600 g) and ethyl acetate (500 mL). The upper organic layer was separated. The aqueous layer was extracted twice with ethyl acetate (500 mL). The combined organic extracts were washed with 23% (w / w) NaCl solution. The organic layer was separated, dried over Na ₂ SO ₄ and filtered. The solution was concentrated to 21.5 g of oil, dissolved in heptane (150 mL) and stirred slowly (crystals formed after 10 minutes). The slurry was stirred at ambient temperature for 15 hours, filtered and washed with ambient heptane (20 mL). The desired product was obtained as white crystals (dried to a constant weight of 12.3 g).

(D) Preparation of (2S) -4-N-carbonylbenzyloxy-L-valinyloxy-2-stearoyloxymethylbutyltoluenesulfonate

Example 31 The product of step (c) (11.91 g, 0.022 mol) was placed in a 250 mL shaker bottle. THF (120 mL) and RaNi (17.8 g) were added. The reaction was pressurized to 4 atm H _2. The reaction was shaken for 1.5 hours. The reaction was filtered and washed with 20 mL THF. The filtrate was diluted with CH ₂ Cl ₂ (100 mL), dried over Na ₂ SO ₄ , filtered and washed with CH ₂ Cl ₂ (25 mL). The filtrate was placed in a 500 mL three-necked flask equipped with a magnetic stir bar and N ₂ inlet. N-Cbz-L-valine (13.88 g, 0.055 mol), 1,3-dicyclohexylcarbodiimide (11.37 g, 0.055 mol) and 4-dimethylaminopyridine (0.40 g, 0.003 mol) And the reaction was stirred for 1 hour. The reaction mixture became heterogeneous after a few minutes. The reaction was filtered and washed with CH ₂ Cl ₂ (50 mL). The filtrate was diluted with ethyl acetate (600 mL) and washed twice with 7% (w / w) NaHCO ₃ solution (100 mL). The organic layer was then washed with 5% (w / w) KH ₂ PO ₄ solution (100 mL). The organic layer was washed with 7% (w / w) NaHCO ₃ solution (100 mL), then dried over MgSO ₄ and filtered. The solution was concentrated to 19.46 g of an oily solid. This solid was dissolved in 30 mL of 8: 2 hexane: ethyl acetate and chromatographed in two portions. Each half was chromatographed on a flash 40M silica gel cartridge (32-63 μm, 60 g silica, 90 g 4.0 × 15.0 cm) and eluted with 8: 2 hexane: ethyl acetate at 25 ml / min. A 25 ml fraction was collected. Fractions were analyzed by TLC. In the first analysis fractions 10-22 contained pure product and in the second analysis fractions 9-26 contained pure product. These fractions were combined and concentrated to give the desired product as a clear viscous oil (12.58 g).

(E) Production of 6-benzyloxy-2-aminopurine

60% sodium hydride in mineral oil (2.36 g, 0.059 mol) was placed in a 500 mL three-necked flask equipped with a magnetic stir bar, temperature probe, condenser, and N ₂ inlet. Toluene (250 mL) was added. Benzyl alcohol (50 mL) was added dropwise over 30 minutes. After the addition of benzyl alcohol, the reaction was stirred for 10 minutes. Then 6-chloro-2-aminopurine (5.00 g, 0.029 mol) was added and the reaction mixture was heated to reflux (115 ° C.) for 4.5 hours. The reaction mixture was filtered hot through a coarse glass sinter funnel to give a wet off-white solid (11.65 g). The wet solid was triturated with CH ₂ Cl ₂ (100 mL) and water (100 mL). After stirring for 10 minutes, the solid dissolved. The aqueous layer was separated and the pH was lowered to 9 with 6M HCl over 3 minutes. A white solid precipitate formed. The slurry was filtered, washed with water (50 mL) and dried to constant weight (under vacuum at 50 ° C.) to give the desired product as off-white crystals (5.15 g).

(F) Production of (R) -9-[(2-stearoyloxymethyl) -4- (N-benzyloxycarbonyl-L-valyloxy) butyl] guanine

The product of Example 31 step (e) (2.40 g, 0.0099 mol) was placed in a 100 mL round bottom flask equipped with a magnetic stir bar and N ₂ inlet. DMF (6 mL) and potassium carbonate (6.27 g) were added. The mixture was stirred at room temperature for 30 minutes. The product of Example 31 step (d) (7.02 g, 0.0091 mol) was dissolved in DMF (21 mL) and added to the mixture. The flask was immersed in a 70 ° C. oil bath and stirred for 24 hours. The reaction was cooled to ambient temperature and poured into a 500 mL separatory funnel containing ethyl acetate (135 mL) and 5% (w / w) KH ₂ SO ₄ solution (135 mL). The topmost organic layer was retained and washed with 7% (w: w) NaHCO ₃ solution (100 mL). The organic layer was dried over MgSO ₄ and filtered. The solution was concentrated to 9.79 oily solid. This was triturated with 50 mL of 1: 1 hexane: ethyl acetate, filtered and concentrated to 9.10 g of a yellow oil. This oil was dissolved in 1/1 hexane-ethyl acetate (20 mL) and chromatographed on a flash 40M silica gel cartridge (32-63 μm, 60Å silica 90 g 4.0 cm × 15.0 cm) to give 6: 4 hexane. Elution with ethyl acetate at 25 ml / min. A 25 ml fraction was collected. These fractions were analyzed by TLC. Fractions 27-92 contained pure product by TLC. Pure fractions were combined and concentrated to give the desired product as an oil (2.95 g).

(G) Production of (R) -9-[(2-stearoyloxymethyl) -4- (L-valyloxy) butyl] guanine

Example 31 The product of step (f) (2.63 g, 0.0031 mol) was dissolved in ethanol (50 mL) and placed in a 500 mL round bottom flask. 10% Pd / C (0.5 g) was slurried in ethanol (20 mL) and added to the flask. The reaction was stirred for 1.5 hours under H ₂ (1 atmosphere from the balloon). The slurry was heated for a while to dissolve the solid, filtered through celite, and washed with hot ethanol (50 mL). The filtrate was concentrated to 1.752 g of a white solid. This solid was dissolved in isopropyl alcohol (10 mL) and isopropyl acetate (42 mL) at 70 ° C. The solution was cooled to 15 ° C. over 2 hours and stirred at 15 ° C. for 12 hours. The solution was cooled to 0 ° C. over 30 minutes and stirred for 1 hour. The slurry was filtered and washed with isopropyl acetate (10 mL). This solid was dried in vacuo at 50 ° C. to give the desired product (0.882 g).

  The mother liquor was concentrated to give 0.55 g of a white solid, which was dissolved at 75 ° C. in isopropyl alcohol (3 mL) and isopropyl acetate (16 mL). The solution was cooled to 15 ° C. over 2 hours, then filtered and dried as described above to give an additional 0.181 g of the desired product.

Other preparations of ethyl 4,4-diethoxy-2-ethoxycarbonylbutyrate To a suspension of sodium ethoxide (20 g, 0.294 mol) in dimethylformamide (68 g) was added diethyl malonate (49 g, 0.306 mol). ) Was added over 13 minutes. After the addition was complete, the mixture was heated to 110 ° C. and bromoacetaldehyde diethyl acetal (40 g, 0.203 mol) was added over 1 hour 45 minutes. After the addition was complete, the mixture was heated to 110 ° C. for 7 hours. The reaction mixture was cooled to room temperature, methyl t-butyl ether (160 g) and water (100 g) were added and the mixture was stirred for 15 minutes. The organic layer was separated and treated with 7% aqueous potassium hydroxide (155 g). The layers were separated and the organic layer was washed with water (100 g) followed by brine (60 g). The organic layer was concentrated to give the crude desired product. This crude product is heated at 160-170 ° C. (bath temperature) in a house vacuum (about 45 mmHg) to distill off volatile impurities to give the desired product (43.6 g). It was.

Other Preparations of (R) -9- [4,4-Diethoxy-2- (hydroxymethyl) butyl] guanine In a 100 mL 1-neck flask, the product of Example 30 step (a) (5 g, 0.0145). Mol) was added followed by KOH (2.05 g, 0.0445 mol) in water (20 mL). The mixture was stirred at reflux for 16-20 hours. The pH was then adjusted to 7.0 by adding acetic acid to the reaction mixture (under reflux). The reaction mixture was then cooled to room temperature and stirred for 30 minutes. The resulting precipitate was collected by filtration and washed with water (5 mL). The resulting solid was dried overnight at a temperature not exceeding 50 ° C. to give the desired product (4.45 g).

Another Purification Method for (R) -9- [4-Hydroxy) -2- (stearoyloxymethyl) butyl] guanine as (S)-(+)-camphorsulfonate salt Example 14 in a 250 mL round bottom flask The product of step (i) (13.0 g) and (1S)-(+)-10-camphor sulfonic acid (5.85 g) were added. Heptane (50 mL) was added and the mixture was stirred for 15 minutes. Then tetrahydrofuran (THF; 50 mL) was added and the mixture was stirred for 5 hours. The resulting precipitate was collected by filtration and washed with heptane (100 mL). The resulting solid was vacuum dried at 45 ° C. to give the desired product (11.3 g). HPLC analysis of the product showed 98.76% ee.

の製造
５０ｍＬ容丸底フラスコに、実施例１４工程（ｈ）の生成物（１.０ｇ、１.７ミリモル）、ＴＨＦ（２０ｍＬ）、Ｈ_２Ｏ（１ｍＬ）およびアンバーライト１５レジン（１.０ｇ）を入れた。ついで、この溶液を６５℃に３時間加熱した。ついで、この溶液を濾過し、レジンをＴＨＦ（２×１０ｍＬ）で洗浄した。ついで、溶媒を真空除去して所望の生成物（０.７４ｇ、８４％）を得た。

A 50 mL round bottom flask was charged with the product of Example 14 step (h) (1.0 g, 1.7 mmol), THF (20 mL), H ₂ O (1 mL) and Amberlite 15 resin (1.0 g). ) The solution was then heated to 65 ° C. for 3 hours. The solution was then filtered and the resin was washed with THF (2 × 10 mL). The solvent was then removed in vacuo to give the desired product (0.74 g, 84%).

Other Preparation of (R) -9- [4-Hydroxy) -2- (stearoyloxymethyl) butyl] guanine Into a 100 mL round bottom flask was added the product of Example 14 step (h) (2.45 g, 4 .14 mmol), THF (25 mL), H ₂ O (1 mL) and Amberlite 15 resin (2.5 g). The solution was then heated to 65 ° C. for 3 hours. The solution was then hot filtered and the resin was washed with THF (2 × 15 mL). The crude aldehyde solution was cooled to room temperature and a solution of borane t-butylamine complex (0.3 g, 3.45 mmol) in THF / H ₂ O (1/20 mL) was added dropwise to the aldehyde solution. It was. The solution was stirred at room temperature for 1.5 hours and then quenched with H ₂ O (100 mL). After stirring for an additional 30 minutes at room temperature, the precipitate was isolated by filtration and dried to give the desired product (1.00 g, 47%).

(R) -9- [4- (N-Benzyloxycarbonyl-L-valyloxy) -2- (stearoyloxymethyl) butyl] guanine other process (a) N-carbobenzyloxy-L-valine anhydride A reactor of dicyclohexylcarbodiimide (5 kg, 24 mol) in acetonitrile (17.5 kg) was charged with a solution of N-carbobenzyloxy-L-valine (12.5 kg, 50 mol) in acetonitrile (200 kg). added. The mixture was stirred at 5 ± 5 ° C. for 6 hours and the resulting solid was filtered off. The filtrate was concentrated in vacuo at a temperature not exceeding 45 ° C. and the residue was dissolved in toluene (50 kg) at 40 ° C. Heptane (50 kg) was added and the mixture was cooled to 15 ± 5 ° C. The precipitate was filtered off and dried to give the desired product (10.2 kg).

(B) (R) -9- [4- (N-benzyloxycarbonyl-L-valyloxy) -2- (stearoyloxymethyl) butyl] guanine (R)-(-[4-hydroxy-2- (stearoyloxy) Methyl) butyl] guanine (5.2 kg, 10 mol), N-CBZ-L-valine anhydride (6.3 kg, 13 mol), 4-dimethylaminopyridine (60 g, 0.5 mol) and tetrahydrofuran (67 kg) Was stirred for 2-4 hours at 25 ± 5 ° C. Water (2 kg) was added and the mixture was concentrated in vacuo at a temperature not exceeding 45 ° C. The residue was dissolved in ethyl acetate (58 kg) and 10% Extracted with aqueous sodium bicarbonate (2 × 50 kg) and water (1 × 50 kg) The ethyl acetate solution was concentrated in vacuo and the residue was dissolved in methanol (20 kg) at 50 ± 5 ° C. . The solution was cooled to 20 ± 5 ° C. and diluted with acetonitrile (50 kg) and water (3 kg). The precipitate was filtered off to afford the desired product was dried under vacuum (5.3 kg).

Other Preparations of (R) -9- [4,4-Diethoxy-2- (stearoyloxymethyl) butyl] guanine Stearic acid (1.05 g) and N-methyl in THF (13 mL) at 0-4 ° C. To a stirred solution of morpholine (0.62 g) was added a solution of p-tosyl chloride (0.67 g) in THF (2 mL) at -3 to -4 ° C. The mixture was stirred at room temperature for 3 hours. The product of Example 14 step (g) (1.0 g) and 4-dimethylaminopyridine (75 mg) were added and the slurry was stirred at room temperature for 5 days and quenched with water (135 mL). The mixture was stirred overnight and the precipitate was filtered and washed with water. The wet filter cake was dried under vacuum (40 ° C.) to give the desired product (1.3 g) as a light yellow powder.

Other Preparations of (R) -9- [4,4-Diethoxy-2- (hydroxymethyl) butyl] guanine Example 30 The product of step (a) (10.0 g, 29.1 mmol) was added to water ( To a solution of sodium hydroxide (2.33 g, 5.82 mmol) in 200 mL). To this suspension was added a solution of trimethylamine (6.61 mL, 43.6 mmol of a 40 wt% solution in water). The heterogeneous solution was stirred overnight at room temperature. The reaction was diluted with water (50 mL) and then extracted with ethyl acetate (200 mL). A saturated solution of ammonium sulfate (300 mL) was added to the aqueous layer. The mixture was stirred at room temperature for 30 hours and the resulting precipitate was filtered. The filter cake was washed with ethyl acetate (100 mL). The product was dried in a vacuum oven (high home vacuum, 45 ° C.) overnight to give the desired product (7.88 g).

Formulation Example A
Tablet formulation The following ingredients are screened through a 0.15 mm sieve and dry mixed.
10 g (R) -9- [2- (stearoyloxymethyl) -4- (L-variable)
Luoxy) butyl] guanine 40 g Lactose 49 g Crystalline cellulose 1 g Magnesium stearate Compress the mixture using a tableting machine to give tablets containing 250 mg of active ingredient.

Formulation Example B
Enteric Tablet Formulation The tablets of Example A are spray coated with a solution containing the following ingredients in a tablet coater.
120 g Ethylcellulose 30 g Propylene glycol 10 g Sorbitan monooleate Up to 1000 ml Distilled water

Formulation Example C
Controlled release preparation 50 g (R) -9- [2- (stearoyloxymethyl) -4- (L-Ba
Ryloxy) butyl] guanine 12 g Hydroxypropyl methylcellulose (Methocel K15)
4.5 g Lactose is dry mixed and granulated with povidone aqueous paste. Magnesium stearate (0.5 g) is added and the mixture is compressed in a tableting machine to 13 mm diameter tablets containing 500 mg of active ingredient.

Formulation Example D
Soft capsule 250 g (R) -9- [2- (stearoyloxymethyl) -4- (L-Ba
Ryloxy) butyl] guanine 100 g lecithin 100 g peanut oil The compound of the invention is dispersed in lecithin and peanut oil and filled into soft gelatin capsules.

Biological Example 1
Bioavailability test in rats The bioavailability of the compounds of the present invention was compared to the parent compound H2G and other H2G derivatives in a rat model. The compounds of the invention and comparative compounds are administered orally (by catheter to the stomach) to 3 individually weighed animals and are aqueous (depending on the solubility of the test compound components) Examples 4, 5, Comparative Examples 1-3, 5, 8) Vehicle, peanut oil (Comparative Examples 4, 9, 10) Vehicle or propylene glycol (Examples 1-3, 6-12, 17, Comparative Examples 6, 7 ) 0.1 mmol / kg of prodrug dissolved in vehicle was given. From 5 hours after dosing to about 17 hours after dosing, animals were fasted and maintained in a metabolic cage. Urine was collected for 24 hours after administration and frozen until analysis. U2H in urine was analyzed using the HPLC / UV assay of Stahle and Oberg, Antimicrob Agents Chemother. 36No2, 339-342 (1992) as follows: Dilute 1: 100 in distilled water H ₂ O, centrifuge at 3000 rpm for 10 minutes and filter through Amicon filter. Two 30 μl samples were HPLC columns (Zorbax SB-C18; 75 × 4.6 mm; 3.5 microns; mobile phase 0.05M NH ₄ PO ₄ , 3-4% methanol, pH 3.3-3.5; Chromatography on 5 ml / min; H2G retention time at 254 mm, MeOH 4%, pH 3.33, ˜12.5 min). Bioavailability was calculated as an H2G measurement collected from each animal averaged over at least 3 animals, each individually weighing 4 animals injected jugularly at 0.1 mmol / kg H2G in Ringer's buffer vehicle. Was expressed as a percentage of the average 24-hour urine H2G recovered from a group of rats that were measured and analyzed as described above.

  Comparative Example 1 (H2G) was from the same batch used for the manufacture of Examples 1-12. The preparation of Comparative Example 2 (Mono Val-H2G) and Comparative Example 3 (DiVal-H2G) is shown in Example 21 and Example 23. Comparative Example 4 (distearoyl H2G) was prepared by diesterification of unprotected H2G under esterification conditions comparable to Step 2 of Example 1. Comparative Examples 5 and 8 (Val / AcH2G) were prepared in the same manner as Example 4 using acetic anhydride and related monovaline H2G. Comparative Example 6 (Ala / stearoyl H2G) was prepared in the same manner as Example 6 using Nt-Boc-L-alanine in Step 4. Comparative Example 7 (Gly / decanoyl) was prepared in the same manner as Example 5 using the Step 1 intermediate prepared with Nt-Boc-L-glycine. The preparation of Comparative Examples 9 and 10 is shown in Examples 24 and 25, respectively. The results are shown in Table 2 below.

Comparison of bioavailability between the compound of the present invention and the compound of the comparative example is
And it gives rise to significantly greater bioavailability as compared to the diamino acid ester or difatty acid esters special combinations corresponding to the amino acids at R 1 _/ R ₂ fatty acid and R _{1 /} R ₂ in. For example, in this model, the compound of Example 1 exhibits 55% better bioavailability than the corresponding divalin ester of Comparative Example 3. The compound of Example 4 shows 25% better bioavailability than the corresponding divaline ester.

  Further, for example, from the results of Comparative Examples 5, 6 and 7, it is also clear that only certain fatty acids of the present invention result in these dramatic and unpredictable increases in pharmacokinetic parameters in combination with certain amino acids.

Biological Example 2
Plasma Concentration in Rats Plasma concentration assays were performed in male Sprague Dawley derived rats. Prior to dosing, the animals were fasted overnight but freed of water. Each compound to be evaluated was prepared as a solution / suspension in propylene glycol at a concentration corresponding to 10 mg H2G / ml and shaken at room temperature for 8 hours. Groups of rats (at least 4 rats in each group) were given an oral dose of 10 mg / kg (1 ml / kg) of each compound; this dose was administered by gavage. Heparin at selected time points after administration (0.25 hours, 0.5 hours, 1 hour, 1.5 hours, 2 hours, 4 hours, 6 hours, 9 hours, 12 hours, 15 hours and 24 hours after administration) Treated blood samples (0.4 ml / sample) were obtained from the tail vein of each animal. The blood sample was immediately cooled in an ice bath. Within 2 hours of collection, plasma was separated from red blood cells by centrifugation and frozen until time of analysis. The target component was separated from plasma protein using acetonitrile precipitation. After lyophilization and reconstitution, plasma concentrations were measured by reverse phase HPLC with fluorescence detection. Oral uptake of H2G and other test compounds was determined by comparing the H2G area under the curve obtained from oral administration to that obtained from 10 mg / kg intravenous administration of H2G administered to another group of rats. The results are shown in Table 1B above.

Biological Example 3
Bioavailability in monkeys The compounds of Example 1 and Comparative Example 3 (see Biological Example 1 above) were orally administered to cynomolgus monkeys by gavage. These solutions included the following:
Example 1 150 mg dissolved in 6.0 ml propylene glycol, corresponding to 25 mg / kg or 0.0295 mmol / kg.
Comparative Example 3 164 mg dissolved in 7.0 ml water, corresponding to 23.4 mg / kg or 0.0295 mmol / kg.

  Blood samples were collected at 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 10 hours and 24 hours. Plasma was separated by centrifugation at 2500 rpm and inactivated at 54 ° C. for 20 minutes before lyophilization before analysis. Plasma H2G levels were monitored by the HPLC / UV assay of Example 30 above.

  FIG. 1 shows plasma H2G recovery as a function of time. Although no statistically significant conclusions can be drawn from a single animal study, animals given the compounds of the invention are slightly more rapid and slightly larger than H2G compared to animals given other H2G prodrugs. It seems to have experienced exposure.

Biological Example 4
Antiviral active herpes simplex virus-1 (HSV-1) infected mice can be used as an animal model to determine the efficacy of antiviral agents in vivo. Mice intraperitoneally inoculated with HSV-1 1000 times LD ₅₀ were formulated into a formulation containing acyclovir, an anti-herpes agent currently on the market (21 and 83 mg / kg in 2% propylene glycol in sterile water vehicle, 5 times starting 5 hours after inoculation with either the compound of Example 29 (3 times daily, orally) or the compound of Example 29 (21 and 83 mg / kg in 2% propylene glycol in sterile water vehicle, 3 times daily, oral). Administration was continued on consecutive days. Animal survival was assessed daily. The results are shown in FIG. 2. FIG. 2 shows the survival rate plotted against time. In the figure legend, the compound of the invention is labeled Example 29 and acyclovir is labeled ACV. The percentage of HSV-1-infected mice that survived was significantly greater when a given amount of a compound of the invention was administered than when an equivalent amount of acyclovir was administered.

  The foregoing has been set forth merely to illustrate the invention and is not intended to limit the invention to the content disclosed herein. Modifications and changes obvious to those skilled in the art are included within the scope and nature of the invention as defined by the appended claims.

Claims (21)

Formula A:

[Wherein R ₆ and R ₇ are C ₁ -C ₇ alkyl or benzyl, or R ₆ and R ₇ together are —CH ₂ CH ₂ —, —CH ₂ CH ₂ CH ₂ — or _{-CH 2 CH 2 CH 2 CH 2} - to form, _{R 8} is optionally substituted alkyl, saturated or mono-unsaturated _C 1 _{-C 21,} this time, the same or different substituents up to 5 There _OH, C 1 -C _{6 alkyl,} C 1 -C _{6 alkoxy,} C 1 -C ₆ alkoxy _C 1 -C _{6 alkyl,} C 1 -C ₆ alkanoyl, amino, halo, cyano, azido, oxo, mercapto and nitro X ₂ is a leaving group or OH, provided that when R ₆ and R ₇ are —CH ₂ CH ₃ and R ₈ is —CH ₃ , X ₂ is (a) OH, (B) -O-C (= O) -C ( H _{3) 3} or (c) is not a _-O-SO 2 -CH _3] a compound represented by. R ₈ is C ₃ -C ₂₁ saturated or monounsaturated optionally substituted alkyl, and up to 5 similar or different substituents are OH, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy _C 1 -C _{6 alkyl,} C 1 -C ₆ alkanoyl, amino, halo, cyano, azido, oxo, independently selected from mercapto and nitro, a compound according to claim 1. R ₆ and R ₇ are —CH ₃ or —CH ₂ CH ₃ , or R ₆ and R ₇ are taken together to form —CH ₂ CH ₂ —, —CH ₂ CH ₂ CH ₂ — or —CH ₂ CH ₂ CH ₂ CH ₂ - to form, _{R 8} is - (CH _{2) 16} CH _{3, X 2} is halogen, sulfonate leaving group or OH, a compound according to claim 1 or 2. R ₆ and _{R 7} are _-CH 2 CH _{3, R 8} is - (CH _{2) 16} CH _{3, X 2} is tosylate or OH, A compound according to claim 3.   The compound according to claim 1, which is (2S) -2-acetoxymethyl-4,4-diethoxybutyltoluenesulfonate. Formula A:

[Wherein R ₆ and R ₇ are C ₁ -C ₇ alkyl or benzyl, or R ₆ and R ₇ together are —CH ₂ CH ₂ —, —CH ₂ CH ₂ CH ₂ — or _{-CH 2 CH 2 CH 2 CH 2} - to form, _{R 8} is optionally substituted alkyl, saturated or mono-unsaturated _C 1 _{-C 21,} this time, the same or different substituents up to 5 There _OH, C 1 -C _{6 alkyl,} C 1 -C _{6 alkoxy,} C 1 -C ₆ alkoxy _C 1 -C _{6 alkyl,} C 1 -C ₆ alkanoyl, amino, halo, cyano, azido, oxo, mercapto and nitro X ₂ is a leaving group or OH, provided that when R ₆ and R ₇ are —CH ₂ CH ₃ and R ₈ is —CH ₃ , X ₂ is (a) OH, (B) -O-C (= O) -C ( H ₃₎ to a method for preparing a compound represented by ₃ or (c) is not a _-O-SO 2 -CH _3],
(A) Formula:

[Wherein R ₆ and R ₇ are the same as described above] and a compound represented by CH ₂ ═CH—OC (O) R ₈ (wherein R ₈ is the same as described above) in the presence of lipase. formula:

[Wherein R ₆ , R ₇ and R ₈ are the same as defined above], and (b) when X ₂ is desirably a leaving group, optionally in step (a) A process comprising converting a hydroxyl group of a product to a leaving group. Formula B:

[Wherein R ₈ is C ₁ -C ₂₁ saturated or monounsaturated optionally substituted alkyl, wherein up to 5 similar or different substituents are OH, C ₁ -C ₆ alkyl, Independently selected from C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy C ₁ -C ₆ alkyl, C ₁ -C ₆ alkanoyl, amino, halo, cyano, azido, oxo, mercapto and nitro, R ₁₂ is — CH (Ph) ₂ , —C (Ph) ₃ or —Si (t-Bu) (CH ₃ ) ₂ (where Ph is phenyl), X ₂ is a leaving group or OH, provided that R _{When 8} is methyl and X ₂ is OH, R ₁₂ is not —Si (t-Bu) (CH ₃ ) ₂ ]. R ₈ is -CH _{3, X 2} is OH, A compound according to claim 7. R ₈ is -CH _{3, X 2} is a halogen or sulfonate leaving group, compounds of claim 7. R ₈ is -CH _{3, X 2} is tosylate, compound of Claim 7. R ₈ is - (CH _{2) 16} CH _{3, X 2} is OH, A compound according to claim 7. R ₈ is - (CH _{2) 16} CH _{3, X 2} is a halogen or sulfonate leaving group, compounds of claim 7. R ₈ is - (CH _{2) 16} CH _{3, X 2} is tosylate, compound of Claim 7. formula:

[Wherein R ₈ is C ₁ -C ₂₁ saturated or monounsaturated optionally substituted alkyl, wherein up to 5 similar or different substituents are OH, C ₁ -C ₆ alkyl, Independently selected from C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy C ₁ -C ₆ alkyl, C ₁ -C ₆ alkanoyl, amino, halo, cyano, azido, oxo, mercapto and nitro, R ₁₂ is — CH (Ph) ₂ , —C (Ph) ₃ or —Si (t-Bu) (CH ₃ ) ₂ (where Ph is phenyl), X ₂ is a leaving group or OH, provided that R _{When 6} is methyl and X ₂ is OH, R ₁₂ is not —Si (t-Bu) (CH ₃ ) ₂ ],
(A) Formula:

[Wherein R ₁₂ is the same as above] is reacted with CH ₂ ═CH—OC (O) R ₈ (wherein R ₈ is the same as above) in the presence of lipase to give a formula:

[Wherein R ₈ and R ₁₂ are as defined above], and (b) if X ₂ is desirably a leaving group, optionally the hydroxyl of the product of step (a) A process comprising converting a group into a leaving group. R ₈ is - (CH _{2) 16} CH _{3, X 2} is a halogen or sulfonate leaving group, The method of claim 14. R ₈ is - (CH _{2) 16} CH _{3, X 2} is tosylate The method of claim 14. formula:

Wherein X ₂ is a halogen or sulfonate leaving group, P ₁ is an N-protecting group, R ₁₀ is a C _{3 to} C ₂₁ saturated or monounsaturated optionally substituted alkyl, until the same or different substituents _OH, C 1 -C _{6 alkyl,} C 1 -C _{6 alkoxy,} C 1 -C ₆ alkoxy _C 1 -C _{6 alkyl,} C 1 -C ₆ alkanoyl, amino, halo, cyano And R ₁₁ is isopropyl or isobutyl, independently selected from azide, oxo, mercapto and nitro. An X ₂ is p- _{toluenesulfonyloxy, R 10} is - (CH _{2) 16} is CH _3, A compound according to claim 17. X ₂ is p- _{toluenesulfonyloxy, R 10} is - a _{(CH 2) 16 CH 3,} R 11 is isopropyl, _{P 1} is benzyloxycarbonyl, A compound according to claim 17. formula:

Wherein X ₂ is a halogen or sulfonate leaving group, R ₁₀ is a C ₃ -C ₂₁ saturated or monounsaturated optionally substituted alkyl, wherein up to 5 similar or different substituents There _OH, C 1 -C _{6 alkyl,} C 1 -C _{6 alkoxy,} C 1 -C ₆ alkoxy _C 1 -C _{6 alkyl,} C 1 -C ₆ alkanoyl, amino, halo, cyano, azido, oxo, mercapto and nitro Is independently selected from]. An X ₂ is p- _{toluenesulfonyloxy, R 10} is - (CH _{2) 16} is CH _3, A compound according to claim 20.

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