Antiviral nucleoside reverse transcriptase inhibitor

Abstract

The present invention relates to a compound of a novel antiviral nucleoside reverse transcriptase inhibitor, a pharmaceutical composition containing the compound, and preparation and application of the compound. Specifically, disclosed in the present invention are a fused pyrimidine compound as represented by formula (I) and a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative of the compound. The compound of the present invention can be used for treatment and/or prevention of infectious diseases caused by viruses, such as human immunodeficiency virus (HIV), hepatitis B virus (HBV). ##STR00001##

Claims

1. A compound of Formula (I-1): ##STR00074## wherein, R.sup.4 is optionally substituted C.sub.1-C.sub.6 alkyl; Y is selected from the following structures: ##STR00075## wherein, R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.3-C.sub.7 carbocyclyl as valency permits; m is 2 or 3; n is 13 or 15; or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, or an isotopic variant thereof.

2. The compound of claim 1, wherein R.sup.4 is methyl.

3. The compound of claim 1, wherein R.sup.7 is selected from the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

4. The compound of claim 1, wherein m is 2.

5. The compound of claim 1, wherein n is 15.

6. The compound of claim 1, wherein the compound has a chirality at the phosphorus atom, and its configuration is selected from S-configuration, R-configuration or the mixture of S-configuration and R-configuration.

7. The compound of claim 1, which is selected from the following compounds: ##STR00076## ##STR00077## ##STR00078## ##STR00079## ##STR00080## ##STR00081## ##STR00082## ##STR00083## ##STR00084## ##STR00085## ##STR00086## or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, or an isotopic variant thereof.

8. The compound of claim 1, wherein R.sup.7 is selected from the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, neopentyl, cyclopentyl, and cyclohexyl.

9. The compound of claim 1, wherein R.sup.7 is isopropyl.

10. A compound, which is selected from the following compounds: ##STR00087## ##STR00088## or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, or an isotopic variant thereof.

11. A pharmaceutical composition, which comprises the compound of claim 1 or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, or an isotopic variant thereof, and pharmaceutically acceptable excipient(s).

12. The pharmaceutical composition of claim 11, which further comprises other therapeutic agent(s).

13. A kit, comprising: a first container comprising a compound of claim 1 or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, or an isotopic variant thereof; and optionally, a second container comprising other therapeutic agent(s); and optionally, a third container comprising a pharmaceutical excipient for diluting or suspending the compound and/or other therapeutic agent(s).

14. A method of treating viral infections in a subject, comprising administering to the subject the compound of claim 1 or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, or an isotopic variant thereof, wherein the viral infections is caused by human immunodeficiency virus (HIV) or hepatitis B virus (HBV).

Description

SPECIFIC EMBODIMENTS OF THE INVENTION

(1) Compounds

(2) In the present disclosure, “the compound disclosed herein” refers to the following compound of formula (I), a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, a prodrug or an isotopic derivatives thereof.

(3) In one embodiment, the present disclosure relates to a compound of formula (I):

(4) ##STR00004##

(5) wherein,

(6) R.sup.1 is selected from OH or NR.sup.5R.sup.6; wherein R.sup.5 and R.sup.6 are each independently selected from H, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl or optionally substituted C.sub.3-C.sub.6 carbocyclyl, or R.sup.5 and R.sup.6 together with the nitrogen atom to which they are attached form optionally substituted 3- to 6-membered heterocyclyl;

(7) R.sup.2 and R.sup.3 are independently selected from H, NH.sub.2, OH, halogen, CN, NO.sub.2, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.1-C.sub.6 acyl, optionally substituted C.sub.2-C.sub.6 alkenyl, optionally substituted C.sub.2-C.sub.6 alkynyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl, optionally substituted C.sub.3-C.sub.7 heterocyclyl, optionally substituted C.sub.6-C.sub.10 aryl or optionally substituted C.sub.5-C.sub.10 heteroaryl;

(8) R.sup.4 is selected from H, halogen, optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.1-C.sub.6 heteroalkyl;

(9) Y is selected from the following structures:

(10) ##STR00005##

(11) wherein R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.1-C.sub.6 acyl, optionally substituted C.sub.2-C.sub.6 alkenyl, optionally substituted C.sub.2-C.sub.6 alkynyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl, optionally substituted C.sub.3-C.sub.7 heterocyclyl, optionally substituted C.sub.6-C.sub.10 aryl or optionally substituted C.sub.5-C.sub.10 heteroaryl, as valency permits; R.sup.8 and R.sup.9 are independently selected from H, halogen, CN, NO.sub.2, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl or optionally substituted C.sub.3-C.sub.7 carbocyclyl;

(12) m is selected from 2 to 6;

(13) n is selected from 10 to 21;

(14) or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, a prodrug or an isotopic derivatives thereof.

(15) Alternatively, in this embodiment, R.sup.1 is NR.sup.5R.sup.6.

(16) In the above embodiment of R.sup.1, R.sup.5 and R.sup.6 are each independently selected from H, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl or optionally substituted C.sub.3-C.sub.6 carbocyclyl, or R.sup.5 and R.sup.6 together with the nitrogen atom to which they are attached form optionally substituted 3- to 6-membered heterocyclyl; alternatively, R.sup.5 and R.sup.6 are each independently selected from H, optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.1-C.sub.6 heteroalkyl; alternatively, R.sup.5 and R.sup.6 are each independently selected from H or optionally substituted C.sub.1-C.sub.6 alkyl; yet alternatively, R.sup.5 and R.sup.6 are both H.

(17) Alternatively, in this embodiment, R.sup.2 and R.sup.3 are independently selected from H, NH.sub.2, OH, halogen, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.1-C.sub.6 acyl, optionally substituted C.sub.2-C.sub.6 alkenyl, optionally substituted C.sub.2-C.sub.6 alkynyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl, optionally substituted C.sub.3-C.sub.7 heterocyclyl, optionally substituted C.sub.6-C.sub.10 aryl or optionally substituted C.sub.5-C.sub.10 heteroaryl; alternatively, R.sup.2 and R.sup.3 are independently selected from H, NH.sub.2, OH, halogen, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.2-C.sub.6 alkenyl or optionally substituted C.sub.2-C.sub.6 alkynyl; alternatively, R.sup.2 and R.sup.3 are independently selected from H, NH.sub.2, OH, halogen or optionally substituted C.sub.1-C.sub.6 alkyl; alternatively, R.sup.2 and R.sup.3 are independently selected from H, NH.sub.2, OH or halogen; yet alternatively, R.sup.2 and R.sup.3 are independently selected from H, NH.sub.2, OH or F; yet alternatively, R.sup.2 and R.sup.3 is independently selected from H or NH.sub.2; still alternatively, R.sup.2 and R.sup.3 are both H.

(18) Alternatively, in this embodiment, R.sup.4 is selected from optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.1-C.sub.6 heteroalkyl; alternatively, R.sup.4 is optionally substituted C.sub.1-C.sub.6 alkyl; yet alternatively, R.sup.4 is methyl.

(19) Alternatively, in this embodiment, R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.1-C.sub.6 acyl, optionally substituted C.sub.2-C.sub.6 alkenyl, optionally substituted C.sub.2-C.sub.6 alkynyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl or optionally substituted C.sub.3-C.sub.7 heterocyclyl; alternatively, R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl or optionally substituted C.sub.3-C.sub.7 heterocyclyl; alternatively, R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.3-C.sub.7 carbocyclyl;

(20) Yet alternatively, R.sup.7 is selected from the following groups:

(21) methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl;

(22) Most alternatively, R.sup.7 is selected from the following groups:

(23) methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, neopentyl, cyclopentyl and cyclohexyl.

(24) Alternatively, in this embodiment, R.sup.8 and R.sup.9 are independently selected from H, halogen, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl or optionally substituted C.sub.3-C.sub.7 carbocyclyl; alternatively, R.sup.8 and R.sup.9 are independently selected from H, halogen or optionally substituted C.sub.1-C.sub.6 alkyl; yet alternatively, R.sup.8 and R.sup.9 are independently selected from H or optionally substituted C.sub.1-C.sub.6 alkyl; still alternatively, R.sup.8 is H, and R.sup.9 is methyl.

(25) Alternatively, in this embodiment, m is selected from 2, 3, 4 or 5; alternatively, m is selected from 2, 3 or 4.

(26) Alternatively, in this embodiment, n is selected from 11 to 19; yet alternatively, n is selected from 11, 13, 15 or 17.

(27) In another embodiment, the compound of formula (I) is the following compound of formula (II):

(28) ##STR00006##

(29) wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.7, R.sup.8, m and n are as defined above.

(30) In another embodiment, the compound of formula (I) is the following compound of formula (III):

(31) ##STR00007##

(32) wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.7, R.sup.9, m and n are as defined above.

(33) In another embodiment, the present disclosure provides a compound of formula (I-1):

(34) ##STR00008##

(35) wherein,

(36) R.sup.4 is selected from H or optionally substituted C.sub.1-C.sub.6 alkyl;

(37) Y is selected from the following structures:

(38) ##STR00009##

(39) wherein, R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl or optionally substituted C.sub.3-C.sub.7 heterocyclyl, as valency permits;

(40) m is selected from 2 to 6;

(41) n is selected from 10 to 21;

(42) or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, a prodrug or an isotopic derivatives thereof.

(43) In some embodiments, R.sup.4 is selected from H or optionally substituted C.sub.1-C.sub.4 alkyl; alternatively, R.sup.4 is H; alternatively, R.sup.4 is methyl.

(44) In some embodiments, Y is

(45) ##STR00010##
wherein, R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl or optionally substituted C.sub.3-C.sub.7 heterocyclyl, as valency permits; alternatively, wherein R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.3-C.sub.7 carbocyclyl; alternatively, wherein R.sup.7 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl; alternatively, wherein R.sup.7 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, neopentyl, cyclopentyl or cyclohexyl; yet alternatively, wherein R.sup.7 is isopropyl.

(46) In some embodiments, Y is

(47) ##STR00011##
wherein, R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 heteroalkyl, optionally substituted C.sub.3-C.sub.7 carbocyclyl or optionally substituted C.sub.3-C.sub.7 heterocyclyl, as valency permits; alternatively, wherein R.sup.7 is selected from optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.3-C.sub.7 carbocyclyl; alternatively, wherein R.sup.7 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl; alternatively, wherein R.sup.7 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, neopentyl, cyclopentyl or cyclohexyl; yet alternatively, wherein R.sup.7 is isopropyl.

(48) In some embodiments, m is selected from 2 to 6; alternatively, m is selected from 2, 3, 4 or 5; alternatively, m is selected from 2, 3 or 4; yet alternatively, m is 2.

(49) In some embodiments, n is selected from 10 to 21; alternatively, n is selected from 11 to 19; alternatively, n is selected from 11, 13, 15 or 17; yet alternatively, n is 15.

(50) In another embodiment, the compounds described herein (including formulae (I) to (III) and subsets thereof (for example, (I-1))) have a chirality at the phosphorus atom, and the configuration is selected from S-configuration, R-configuration or a mixture of S-configuration and R-configuration.

(51) In another embodiment, the above compound of formulae (I) to (III) is the following compounds:

(52) ##STR00012## ##STR00013## ##STR00014## ##STR00015## ##STR00016## ##STR00017## ##STR00018## ##STR00019## ##STR00020## ##STR00021## ##STR00022## ##STR00023## ##STR00024## ##STR00025## ##STR00026## ##STR00027##

(53) Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various stereoisomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer (such as cis- and trans-isomer), or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses.

(54) Also disclosed herein are all suitable isotopic derivatives of the compounds disclosed herein. An isotopic derivative of a compound disclosed herein is defined as wherein at least one atom is replaced by an atom having the same atomic number but differing in atomic mass from the atomic mass typically found in nature. Examples of isotopes that can be incorporated into compounds disclosed herein include hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine isotopes, such as .sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.17O, .sup.18O, .sup.18F, .sup.31F, .sup.32P, .sup.35S and .sup.36Cl, respectively. Certain isotopic derivatives of the compounds disclosed herein, such as the radioisotopes of .sup.3H and .sup.14C are incorporated are useful in the tissue distribution experiments of drugs and substrates. Tritium, i.e., .sup.3H, and carbon-14, i.e., .sup.14C, are particularly preferred for their ease of preparation and detectability. In addition, substitution with heavier isotopes such as deuterium, i.e., .sup.2H, has therapeutic advantages due to its good metabolic stability, for example, increased half-life in vivo or reduced dosage, and is thus preferable in some cases. Isotopic derivatives of the compounds disclosed herein can be prepared conventionally by the following procedures, for example by descriptive methods or by the preparations described in the Examples below, using appropriate reagents containing appropriate isotopes.

(55) The compounds of the present disclosure or a pharmaceutically acceptable salt thereof may be in an amorphous or crystalline form. Furthermore, the compounds disclosed herein may exist in one or more crystalline forms. Accordingly, the disclosure includes within its scope all amorphous or crystalline forms of the compounds disclosed herein. The term “polymorph” refers to the crystalline form (or its salt, hydrate or solvate) of a compound in a specific crystal packing arrangement. All polymorphs have the same elemental composition. Different crystalline forms usually have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, photoelectric properties, stability, and solubility. Recrystallization solvent, crystallization rate, storage temperature and other factors can cause a crystalline form to dominate. Various polymorphs of the compound can be prepared by crystallization under different conditions.

(56) Those skilled in the art will appreciate that many organic compounds can form complexes with solvents that react in or precipitate or crystallize from the solvent. These complexes are referred to as “solvates.” When the solvent is water, the complex is referred to as a “hydrate.” The disclosure encompasses all solvates of the compounds disclosed herein.

(57) In addition, prodrugs are also included within the context of the present disclosure. The term “prodrug” as used herein refers to a compound which is converted in vivo to an active form thereof having a medical effect by, for example, hydrolysis in blood. Pharmaceutically acceptable prodrugs are described in T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, A.C.S. Symposium Series, Vol. 14, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, and D. Fleisher, S. Ramon and H. Barbra “Improved oral drug delivery: solubility limitations overcome by the use of prodrugs”, Advanced Drug Delivery Reviews (1996) 19(2) 115-130, each is incorporated herein by reference.

(58) A prodrug is any covalently bonded carrier which, when administered to a patient, releases the compound of formula (I) in vivo. Prodrugs are typically prepared by modifying functional groups in such a way that the modification can be produced by conventional operations or cleavage in vivo to yield the parent compound. Prodrugs include, for example, compounds disclosed herein wherein a hydroxy, amino or sulfhydryl group is bonded to any group which, when administered to a patient, can be cleaved to form a hydroxy, amino or sulfhydryl group. Therefore, representative examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of alcohol, mercapto, and amine functional groups of the compound of formula (I). Further, in the case of a carboxylic acid (—COOH), an ester such as a methyl ester, an ethyl ester or the like can be used. The ester itself may be active and/or may hydrolyze under conditions in human bodies. Suitable pharmaceutically acceptable hydrolysable in vivo ester groups include those groups which readily decompose in the human body to release the parent acid or a salt thereof.

(59) Pharmaceutical Compositions, Formulations and Kits

(60) In another aspect, the disclosure provides a pharmaceutical composition comprising a compound of the present disclosure (also referred to as the “active ingredient”) and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises an effective amount of the active ingredient. In certain embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the active ingredient. In certain embodiments, the pharmaceutical composition comprises a prophylactically effective amount of the active ingredient.

(61) A pharmaceutically acceptable excipient for use in the present disclosure refers to a non-toxic carrier, adjuvant or vehicle which does not destroy the pharmacological activity of the compound formulated together. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present disclosure include, but are not limited to, ion exchangers, alumina oxide, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffer substances (such as phosphate), glycine, sorbic acid, potassium sorbate, a mixture of partial glycerides of saturated plant fatty acids, water, salt or electrolyte (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, silica gel, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin.

(62) The present disclosure also includes kits (e.g., pharmaceutical packs). Kits provided may include a compound disclosed herein, other therapeutic agents, and a first and a second containers (eg, vials, ampoules, bottles, syringes, and/or dispersible packages or other materials) containing the compound disclosed herein or other therapeutic agents. In some embodiments, kits provided can also optionally include a third container containing a pharmaceutically acceptable excipient for diluting or suspending the compound disclosed herein and/or other therapeutic agent. In some embodiments, the compound disclosed herein provided in the first container and the other therapeutic agents provided in the second container is combined to form a unit dosage form.

(63) The following formulation examples illustrate representative pharmaceutical compositions that may be prepared in accordance with this disclosure. The present disclosure, however, is not limited to the following pharmaceutical compositions.

(64) Exemplary Formulation 1—Tablets: A compound of the present disclosure may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 0.3-30 mg tablets (0.1-10 mg of active compound per tablet) in a tablet press.

(65) Exemplary Formulation 2—Tablets: A compound of the present disclosure may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 30-90 mg tablets (10-30 mg of active compound per tablet) in a tablet press.

(66) Exemplary Formulation 3—Tablets: A compound of the present disclosure may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 90-150 mg tablets (30-50 mg of active compound per tablet) in a tablet press.

(67) Exemplary Formulation 4—Tablets: A compound of the present disclosure may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 150-240 mg tablets (50-80 mg of active compound per tablet) in a tablet press.

(68) Exemplary Formulation 5—Tablets: A compound of the present disclosure may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 240-270 mg tablets (80-90 mg of active compound per tablet) in a tablet press.

(69) Exemplary Formulation 6—Tablets: A compound of the present disclosure may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 270-450 mg tablets (90-150 mg of active compound per tablet) in a tablet press.

(70) Exemplary Formulation 7—Tablets: A compound of the present disclosure may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 450-900 mg tablets (150-300 mg of active compound) in a tablet press.

(71) Exemplary Formulation 8—Capsules: A compound of the present disclosure may be admixed as a dry powder with a starch diluent in an approximate 1:1 weight ratio. The mixture is filled into 250 mg capsules (125 mg of active compound per capsule).

(72) Exemplary Formulation 9—Liquid: A compound of the present disclosure (125 mg) may be admixed with sucrose (1.75 g) and xanthan gum (4 mg) and the resultant mixture may be blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of microcrystalline cellulose and sodium carboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg), flavor, and color are diluted with water and added with stirring. Sufficient water may then be added to produce a total volume of 5 mL.

(73) Exemplary Formulation 10—Injection: A compound of the present disclosure may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of approximately 5 mg/mL.

(74) Administration

(75) The pharmaceutical composition provided by the present disclosure can be administered by a variety of routes including, but not limited to, oral administration, parenteral administration, inhalation administration, topical administration, rectal administration, nasal administration, oral administration, vaginal administration, administration by implant or other means of administration. For example, parenteral administration as used herein includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, intra-articular administration, intraarterial administration, intrasynovial administration, infrasternal administration, intracerebroventricular administration, intralesional administration, and intracranial injection or infusion techniques.

(76) Generally, the compounds provided herein are administered in an effective amount. The amount of the compound actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

(77) When used to prevent the disorder disclosed herein, the compounds provided herein will be administered to a subject at risk for developing the condition, typically on the advice and under the supervision of a physician, at the dosage levels described above. Subjects at risk for developing a particular condition generally include those that have a family history of the condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition.

(78) The pharmaceutical compositions provided herein can also be administered chronically (“chronic administration”). Chronic administration refers to administration of a compound or pharmaceutical composition thereof over an extended period of time, e.g., for example, over 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc, or may be continued indefinitely, for example, for the rest of the subject's life. In certain embodiments, the chronic administration is intended to provide a constant level of the compound in the blood, e.g., within the therapeutic window over the extended period of time.

(79) The pharmaceutical compositions of the present disclosure may be further delivered using a variety of dosing methods. For example, in certain embodiments, the pharmaceutical composition may be given as a bolus, e.g., in order to raise the concentration of the compound in the blood to an effective level. The bolus dose depends on the target systemic level of the active ingredient passing through the body, e.g., an intramuscular or subcutaneous bolus dose allows a slow release of the active ingredient, while a bolus delivered directly to the veins (e.g., through an IV drip) allows a much faster delivery which quickly raises the concentration of the active ingredient in the blood to an effective level. In other embodiments, the pharmaceutical composition may be administered as a continuous infusion, e.g., by IV drip, to provide maintenance of a steady-state concentration of the active ingredient in the subject's body. Furthermore, in still yet other embodiments, the pharmaceutical composition may be administered as first as a bolus dose, followed by continuous infusion.

(80) The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the compound is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or excipients and processing aids helpful for forming the desired dosing form.

(81) With oral dosing, one to five and especially two to four and typically three oral doses per day are representative regimens. Using these dosing patterns, each dose provides from about 0.01 to about 20 mg/kg of the compound provided herein, with preferred doses each providing from about 0.1 to about 10 mg/kg, and especially about 1 to about 5 mg/kg.

(82) Transdermal doses are generally selected to provide similar or lower blood levels than are achieved using injection doses, generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight.

(83) Injection dose levels range from about 0.1 mg/kg/hour to at least 10 mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to 96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 2 g/day for a 40 to 80 kg human patient.

(84) Liquid forms suitable for oral administration may include a suitable aqueous or nonaqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. Solid forms may include, for example, any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

(85) Injectable compositions are typically based upon injectable sterile saline or phosphate-buffered saline or other injectable excipients known in the art. As before, the active compound in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable excipient and the like.

(86) Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s). When formulated as a ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional dermal penetration ingredients to enhance the stability of the active ingredients or Formulation. All such known transdermal formulations and ingredients are included within the scope provided herein.

(87) The compounds provided herein can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.

(88) The above-described components for orally administrable, injectable or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pa., which is incorporated herein by reference.

(89) The compounds of the present disclosure can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

(90) The present disclosure also relates to the pharmaceutically acceptable formulations of a compound of the present disclosure. In one embodiment, the formulation comprises water. In another embodiment, the formulation comprises a cyclodextrin derivative. The most common cyclodextrins are α-, β- and γ-cyclodextrins consisting of 6, 7 and 8α-1,4-linked glucose units, respectively, optionally comprising one or more substituents on the linked sugar moieties, which include, but are not limited to, methylated, hydroxyalkylated, acylated, and sulfoalkylether substitution. In certain embodiments, the cyclodextrin is a sulfoalkyl ether β-cyclodextrin, e.g., for example, sulfobutyl ether β-cyclodextrin, also known as Captisol. See, e.g., U.S. Pat. No. 5,376,645. In certain embodiments, the formulation comprises hexapropyl-β-cyclodextrin (e.g., 10-50% in water).

(91) Combination Therapy

(92) The compound disclosed herein or its composition can be administered in combination with other therapeutic agents to treat said diseases. Examples of the known therapeutic agent include, but are not limited to:

(93) 1) amprenavir, atazanavir, fosamprenavir, indinavir, lopinavir, ritonavir, nelfinavir, saquinavir, tipranavir, brecanavir, darunavir, TMC-126, TMC-114, mozenavir, JE-2147 (AG1776), L-756423, R00334649, KNI-272, DPC-681, DPC-684, GW640385X, DG17, GS-8374, PPL-100, DG35, and AG1859;

(94) 2) non-nucleoside inhibitors of HIV reverse transcriptase, for example capravirine, emivirine, delaviridine, efavirenz, nevirapine, (+) calanolide A, etravirine, GW5634, DPC-083, DPC-961, DPC-963, WIV-150, TMC-120, rilpivirine, BILR355BS, VPX840773, UK-453061, and RDEA806;

(95) 3) nucleoside inhibitors of HIV reverse transcriptase, for example zidovudine, emtricitabine, didanosine, stavudine (d4T), zalcitabine (ddC), lamivudine (3TC), abacavir, amdoxovir, elvucitabine, alovudine, MIV-210, racivir, D-d4FC, phosphazide, fozivudine tidoxil, apricitibine (AVX754), GS-7340, KP-1461, and fosalvudine tidoxil;

(96) 4) nucleotide inhibitors of HIV reverse transcriptase, for example tenofovir alafenamide and adefovir dipivoxil;

(97) 5) HIV integrase inhibitors, for example curcumin, derivatives of curcumin, chicoric acid, derivatives of chicoric acid, 3,5-dicaffeoylquinic acid, derivatives of 3,5-dicaffeoylquinic acid, aurintricarboxylic acid, derivatives of aurintricarboxylic acid, caffeic acid phenethyl ester, derivatives of caffeic acid phenethyl ester, tyrphostin, derivatives of tyrphostin, quercetin, derivatives of quercetin, S-1360, zintevir, L-870812, and L-870810, raltegravir, elvitegravir, BMS-538158, GSK364735C, BMS-707035, MK-2048, and BA011;

(98) 6) gp41 inhibitors, for example enfuvirtide, sifuvirtide, FB006M, and TRI-1144;

(99) 7) CXCR4 inhibitors, for example AMD-070;

(100) 8) entry inhibitors, for example SP01A;

(101) 9) gp120 inhibitors, for example BMS-488043 or BlockAide/CR;

(102) 10) G6PD and NADH-oxidase inhibitors, for example immunitin;

(103) 11) CCR5 inhibitors, for example aplaviroc, vicriviroc, maraviroc, PRO-140, INCB15050, PF-233798, and CCR5mAb004;

(104) 12) other drugs for treating HIV, for example BAS-100, SPI-452, REP9, SP-01A, TNX-355, DES6, ODN-93, ODN-112, VGX-1, PA-457 (bevirimat), Ampligen, HRG214, Cytolin, VGX-410, KD-247, AMZ0026, CYT99007A-221HIV, DEBIO-025, BAY50-4798, MDX010 (ipilimumab), PBS119, ALG889, and PA-1050040 (PA-040);

(105) 13) interferons, for example pegylated rIFN-alpha 2b, pegylated rIFN-alpha 2a, rIFN-alpha 2b, rIFN-alpha 2a, consensus IFN alpha (infergen), feron, reaferon, intermax alpha, r-IFN-beta, infergen+actimmune, IFN-omega with DUROS, albuferon, locteron, Albuferon, Rebif, Oral interferon alpha, IFNalpha-2b XL, AVI-005, PEG-Infergen, and Pegylated IFN-beta;

(106) 14) ribavirin analogs, for example rebetol, copegus, and viramidine (taribavirin);

(107) 15) NS5b polymerase inhibitors, for example NM-283, valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB1941, XTL-2125, MK-0608, NM-107, R7128 (R4048), VCH-759, PF-868554, and GSK625433;

(108) 16) NS3 protease inhibitors, for example SCH-503034 (SCH-7), VX-950 (telaprevir), BILN-2065, BMS-605339, and ITMN-191;

(109) 17) alpha-glucosidase 1 inhibitors, for example MX-3253 (celgosivir) and UT-231B;

(110) 18) hepatoprotectants, for example IDN-6556, ME3738, LB-84451, and MitoQ;

(111) 19) non-nucleoside inhibitors of HCV, for example benzimidazole derivatives, benzo-1,2,4-thiadiazine derivatives, phenylalanine derivatives, A-831, GS-9190 and A-689; and

(112) 20) other drugs for treating HCV, for example zadaxin, nitazoxanide, BIVN-401 (virostat), PYN-17 (altirex), KPE02003002, actilon, KRN-7000, civacir, GI-5005, ANA-975, XTL-6856, ANA971, NOV-205, tarvacin, EHC-18, NIM811, DEBIO-025, VGX-410C, EMZ-702, AVI4065, Bavituximab, Oglufanide, and VX-497 (merimepodib).

(113) Those other active agents can be administered separately from the composition containing the compound disclosed herein as part of a multiple-dose regimen. Alternatively, those active agents may be part of a single dosage form, mixed with the compound disclosed herein in a single composition. f administered as part of a multiple dosing regimen, the two active agents can be provided simultaneously, sequentially, or separated from each other for a period of time (usually within 5 hours of each other).

(114) Treatment

(115) The present disclosure provides a method of treating and/or preventing viral infections or a method of treating diseases, which comprises the steps of: administering to a subject in need of such treatment the compound disclosed herein, or a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a polymorph, a prodrug or an isotopic derivatives thereof, or pharmaceutical composition disclosed herein.

(116) The compound disclosed herein can treat and/or prevent viral infections, including, but are not limited to human immunodeficiency virus (HIV) infection, hepatitis B virus (HBV) infection.

EXAMPLES

(117) The following examples aim to provide those skilled in the art with a complete disclosure and description of how to perform, prepare, and evaluate the methods and compounds herein, and are intended to merely exemplify the invention and not to limit the scope of the invention as deemed by the inventor.

(118) The Method of Synthesis

(119) The compound disclosed herein can be prepared according to conventional methods in the art, using suitable reagents, raw materials, and purification methods known to those skilled in the art.

(120) The preparation method of the compound of formula (I) disclosed herein is described in more detail below, but these specific methods do not constitute any limitation to the present invention. The compound disclosed herein can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art.

(121) Generally, in the preparation, each reaction is usually carried out in an inert solvent at room temperature to reflux temperature (such as 0° C. to 100° C., preferably 0° C. to 80° C.). The reaction time is usually 0.1 hour to 60 hours, preferably 0.5 to 24 hours.

Example 1

(R)-9-{2-[(dodecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-1)

(122) ##STR00028##

(123) The following synthetic route was used for the synthesis:

(124) ##STR00029##

Step 1: Synthesis of 2-(dodecyldisulfanyl)ethanol (Compound 1)

(125) To a reaction flask were added 2-mercaptoethanol (780 mg, 10 mmol), and n-dodecanethiol (2.02 g, 10 mmol), which were dissolved in 35 ml anhydrous methanol and 35 ml anhydrous dichloromethane. Pyridine (1.58 g, 20 mmol) was added to the mixture. Under the nitrogen protection, elemental iodine (2.54 g, 10 mmol) was added to the mixture in portions, and the reaction was stirred at room temperature for 2-5 hours after the addition was completed. Insoluble material was filtered off upon TLC detection showed the reaction was completed, and the filtrate was concentrated to dryness. The residue was dissolved in a small amount of dichloromethane, washed with saturated brine, concentrated, purified by column chromatography, and dried in vacuo to afford 1.13 g of a product in a yield of 40.6%.

Step 2: Synthesis of (R)-(1-(6-((dimethylamino)methyleneamino)-9H-purin-9-yl)propan-2-yloxy)methylphosphonic dichloride (Compound 2)

(126) To a reaction flask were added (R)-(1-(6-((dimethylamino)methyleneamino)-9H-purin-9-yl)propan-2-yloxy)methylphosphonic acid (tenofovir, 500 mg, 1.74 mmol) and anhydrous DMF (153 mg, 2.1 mmol), which were dissolved by adding 10 ml anhydrous dichloromethane. Under the nitrogen protection, 2 M oxalyl chloride (4.35 ml, 8.7 mmol) was added dropwise at room temperature. After the addition, the reaction was stirred for 3 hours until the reaction became clear, which was concentrated to remove the solvent and excess oxalyl chloride to afford 658 mg of a product in a yield of 100%, which was used in the next step directly without purification.

Step 3: Synthesis of (R)-9-{2-[(dodecyldithioethyl)phosphorylmethoxy]propyl}adenine (Compound 3)

(127) To a reaction flask was added Compound 2 (658 mg, 1.74 mmol), which was dissolved by adding 10 ml anhydrous dichloromethane. Under the nitrogen protection, the reaction was cooled to 0° C., and Compound 1 (584.2 mg, 2.1 mmol) and pyridine (826 mg, 10.44 mmol) in anhydrous dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was reacted at the low temperature for 10 minutes and warmed to rt and reacted for 2 hours. Upon MS monitoring showed the reaction was completed, the reaction was quenched by adding 1 M dilute hydrochloric acid (10 ml), and stirred overnight at room temperature. The organic phase was separated upon MS monitoring showed the reaction was completed. The aqueous phase was extracted 2-3 times with dichloromethane. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 758 mg of product in a yield of 79.6%. LC-MS(APCI): m/z=548.3 (M+1).sup.+.

Step 4: Synthesis of (R)-9-{2-[(dodecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-1)

(128) To a reaction flask were added Compound 3 (228 mg, 0.417 mmol), chloromethyl isopropyl carbonate (316.7 mg, 2.08 mmol), potassium carbonate (287.5 mg, 2.08 mmol) and potassium iodide (34.8 mg, 0.21 mmol). 8 ml DMF was added into the mixture, which was heated to 60° C. overnight. The reaction was cooled to rt upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 103 mg of product in a yield of 37.2%. LC-MS(APCI): m/z=664.3 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.33 (s, 1H), 8.01 (s, 1H), 6.03 (s, 2H), 5.61 (m, 2H), 4.92 (m, 1H), 4.40-4.12 (m, 4H), 3.92 (m, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.85 (dt, J=19.2, 6.7 Hz, 2H), 2.65 (dd, J=11.8, 7.0 Hz, 2H), 1.64 (s, 2H), 1.33-1.22 (m, 27H), 0.84 (t, J=6.6 Hz, 3H).

Example 2

(R)-9-{2-[(tetradecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-2)

(129) ##STR00030##

(130) The following synthetic route was used for the synthesis:

(131) ##STR00031##

Step 1: Synthesis of 2-(tetradecyldisulfanyl)ethanol (Compound 4)

(132) To a reaction flask were added 2-mercaptoethanol (780 mg, 10 mmol), and n-tetradecanethiol (2.3 g, 10 mmol), which were dissolved in 35 ml anhydrous methanol and 35 ml anhydrous dichloromethane. Pyridine (1.58 g, 20 mmol) was added to the mixture. Under the nitrogen protection, elemental iodine (2.54 g, 10 mmol) was added to the mixture in portions, and the reaction was stirred at room temperature for 2-5 hours after the addition was completed. Insoluble material was filtered off upon TLC detection showed the reaction was completed, and the filtrate was concentrated to dryness. The residue was dissolved in a small amount of dichloromethane, washed with saturated brine, concentrated, purified by column chromatography, and dried in vacuo to afford product 1.55 g in a yield of 50.6%.

Step 2: Synthesis of (R)-9-{2-[(tetradecyldisulfanylethyl)phosphorylmethoxy]propyl}adenine (Compound 5)

(133) To a reaction flask was added Compound 2 (658 mg, 1.74 mmol), which was dissolved in 10 ml anhydrous dichloromethane Under the nitrogen protection, the reaction was cooled to 0° C., and a solution of Compound 4 (643 mg, 2.1 mmol) and pyridine (826 mg, 10.44 mmol) in anhydrous dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was reacted at the low temperature for 10 minutes and warmed to rt and reacted for 2 hours. Upon MS monitoring showed the reaction was completed, 1 M dilute hydrochloric acid (10 ml) was added to quench the reaction, which was stirred at room temperature overnight. The organic phase was separated upon MS monitoring showed the reaction was completed. The aqueous phase was extracted 2-3 times with dichloromethane. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 732 mg of product in a yield of 73.1%. LC-MS(APCI): m/z=576.2 (M+1).sup.+.

Step 3: Synthesis of (R)-9-{2-[(tetradecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-2)

(134) To a reaction flask were added Compound 5 (300 mg, 0.52 mmol), chloromethyl isopropyl carbonate (395.3 mg, 2.6 mmol), potassium carbonate (359.3 mg, 2.6 mmol) and potassium iodide (43.2 mg, 0.26 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 124 mg of product in a yield of 34.5%. LC-MS(APCI): m/z=692.7 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.36 (s, 1H), 8.04 (s, 1H), 6.06 (s, 2H), 5.65 (dd, J=28.2, 8.9 Hz, 2H), 4.92 (m, 1H), 4.40-4.12 (m, 4H), 3.96 (m, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.89 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.64 (s, 2H), 1.33-1.25 (m, 31H), 0.87 (t, J=6.6 Hz, 3H).

Example 3

(R)-9-{2-[(hexadecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-3)

(135) ##STR00032##

(136) The following synthetic route was used for the synthesis:

(137) ##STR00033##

Step 1: Synthesis of 2-(hexadecyldisulfanyl)ethanol (Compound 6)

(138) To a reaction flask were added 2-mercaptoethanol (780 mg, 10 mmol), and n-hexadecanethiol (2.58 g, 10 mmol), which were dissolved in 35 ml anhydrous methanol and 35 ml anhydrous dichloromethane. Pyridine (1.58 g, 20 mmol) was added to the mixture. Under the nitrogen protection, elemental iodine (2.54 g, 10 mmol) was added to the mixture in portions, and the reaction was stirred at room temperature for 2-5 hours after the addition was completed. Insoluble material was filtered off upon TLC detection showed the reaction was completed, and the filtrate was concentrated to dryness. The residue was dissolved in a small amount of dichloromethane, washed with saturated brine, concentrated, purified by column chromatography, and dried in vacuo to afford product 1.89 g in a yield of 56.6%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)phosphorylmethoxy]propyl}adenine (Compound 7)

(139) To a reaction flask was added Compound 2 (658 mg, 1.74 mmol), which was dissolved in 10 ml anhydrous dichloromethane Under the nitrogen protection, the reaction was cooled to 0° C., and a solution of Compound 6 (701.9 mg, 2.1 mmol) and pyridine (826 mg, 10.44 mmol) in anhydrous dichloromethane was added to the reaction dropwise. After the addition, the reaction was reacted at the low temperature for 10 minutes and warmed to rt and reacted for 2 hours. Upon MS monitoring showed the reaction was completed, 1 M dilute hydrochloric acid (10 ml) was added to quench the reaction, which was stirred at room temperature overnight. The organic phase was separated upon MS monitoring showed the reaction was completed, and the aqueous phase was extracted 2-3 times with dichloromethane. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 895 mg of product in a yield of 85.2%. LC-MS(APCI): m/z=604.5 (M+1).sup.+.

Step 3: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-3)

(140) To a reaction flask were added Compound 7 (140 mg, 0.23 mmol), chloromethyl isopropyl carbonate (176.4 mg, 1.16 mmol), potassium carbonate (160.3 mg, 1.16 mmol) and potassium iodide (19.9 mg, 0.12 mmol). 5 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 45 mg of product in a yield of 27.2%. LC-MS(APCI): m/z=720.3 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (dd, J=28.2, 8.9 Hz, 2H), 4.92 (m, 1H), 4.40-4.12 (m, 4H), 3.92 (m, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.87 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.64 (s, 2H), 1.33-1.22 (m, 35H), 0.87 (t, J=6.6 Hz, 3H).

Example 4

(R)-9-{2-[(R)-(hexadecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-3-1)

(141) ##STR00034##

(142) Compound T-3-1 was prepared according to the method described in Example 3, and was obtained by chiral column separation.

Example 5

(R)-9-{2-[(S)-(hexadecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-3-2)

(143) ##STR00035##

(144) Compound T-3-2 was prepared according to the method described in Example 3, and was obtained by chiral column separation.

Example 6

(R)-9-{2-[(octadecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-4

(145) ##STR00036##

(146) The following synthetic route was used for the synthesis:

(147) ##STR00037##

Step 1: Synthesis of 2-(octadecyldisulfanyl)ethanol (Compound 8)

(148) To a reaction flask were added 2-mercaptoethanol (780 mg, 10 mmol), and n-octadecanethiol (2.86 g, 10 mmol), which were dissolved in 35 ml anhydrous methanol and 35 ml anhydrous dichloromethane. Pyridine (1.58 g, 20 mmol) was added to the mixture. Under the nitrogen protection, elemental iodine (2.54 g, 10 mmol) was added to the mixture in portions, and the reaction was stirred at room temperature for 2-5 hours after the addition was completed. Insoluble material was filtered off upon TLC detection showed the reaction was completed, and the filtrate was concentrated to dryness. The residue was dissolved in a small amount of dichloromethane, washed with saturated brine, concentrated, purified by column chromatography, and dried in vacuo to afford product 2.38 g in a yield of 65.7%.

Step 2: Synthesis of (R)-9-{2-[(octadecyldisulfanylethyl)phosphorylmethoxy]propyl}adenine (Compound 9)

(149) To a reaction flask was added Compound 2 (658 mg, 1.74 mmol), which was dissolved in 10 ml anhydrous dichloromethane Under the nitrogen protection, the reaction was cooled to 0° C., and a solution of Compound 8 (761 mg, 2.1 mmol) and pyridine (826 mg, 10.44 mmol) in anhydrous dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was reacted at the low temperature for 10 minutes and warmed to rt and reacted for 2 hours. Upon MS monitoring showed the reaction was completed, and 1 M dilute hydrochloric acid (10 ml) was added to quench the reaction, which was stirred at room temperature overnight. The organic phase was separated upon MS monitoring showed the reaction was completed. The aqueous phase was extracted 2-3 times with dichloromethane. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 1.02 g of product in a yield of 92.9%. LC-MS(APCI): m/z=632.8 (M+1).sup.+.

Step 3: Synthesis of (R)-9-{2-[(octadecyldisulfanylethyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-4)

(150) To a reaction flask were added Compound 9 (640 mg, 1.01 mmol), chloromethyl isopropyl carbonate (770.5 mg, 5.07 mmol), potassium carbonate (700.7 mg, 5.07 mmol) and potassium iodide (84.6 mg, 0.51 mmol) 15 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 239 mg of product in a yield of 31.7%. LC-MS(APCI): m/z=748.6 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.36 (s, 1H), 8.05 (s, 1H), 6.06 (s, 2H), 5.61 (m, 2H), 4.92 (m, 1H), 4.40-4.12 (m, 4H), 3.92 (m, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.89 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.66 (s, 2H), 1.33-1.22 (m, 39H), 0.89 (t, J=6.6 Hz, 3H).

Example 7

(R)-9-{2-[(hexadecyldisulfanylethyl)(methylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-5)

(151) ##STR00038##

(152) The following synthetic route was used for the synthesis:

(153) ##STR00039##

Step 1: Synthesis of Chloromethyl Methyl Carbonate (Compound 10)

(154) To a reaction flask were added chloromethyl chloroformate (1.5 g, 11.72 mmol) and methanol (375 mg, 11.72 mmol), which were dissolved in 10 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (0.93 g, 11.72 mmol) was added to the reaction dropwise slowly. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 1.32 g in a yield of 90.8%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(methylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-5)

(155) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 10 (513.8 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 83 mg of product in a yield of 14.5%. LC-MS(APCI): m/z=692.1 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.63 (dd, J=28.2, 8.9 Hz, 2H), 4.92 (m, 1H), 4.40-4.12 (m, 3H), 3.92 (m, 2H), 3.84 (s, 3H), 3.68 (t, J=10.0 Hz, 1H), 2.86 (dt, J=19.2, 6.7 Hz, 2H), 2.64 (dd, J=11.8, 7.0 Hz, 2H), 1.64 (s, 2H), 1.33-1.22 (m, 29H), 0.87 (t, J=6.6 Hz, 3H).

Example 8

(R)-9-{2-[(hexadecyldisulfanylethyl)(ethylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-6)

(156) ##STR00040##

(157) The following synthetic route was used for the synthesis:

(158) ##STR00041##

Step 1: Synthesis of Chloromethyl Ethyl Carbonate (Compound 11)

(159) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and ethanol (1.08 g, 23.45 mmol), which were dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added to the reaction dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 2.51 g in a yield of 77.5%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(ethylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-6)

(160) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 11 (571.9 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 99 mg of product in a yield of 16.9%. LC-MS(APCI): m/z=706.4 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.32 (s, 1H), 8.01 (s, 1H), 6.06 (s, 2H), 5.61 (dd, J=28.2, 8.9 Hz, 2H), 4.91 (m, 1H), 4.40-4.12 (m, 4H), 3.92 (m, 3H), 3.68 (t, J=10.0 Hz, 1H), 2.88 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.68 (t, J=9.5 Hz, 3H), 1.64 (s, 2H), 1.33-1.22 (m, 29H), 0.86 (t, J=6.6 Hz, 3H).

Example 9

(R)-9-{2-[(hexadecyldisulfanylethyl)(propylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-7)

(161) ##STR00042##

(162) The following synthetic route was used for the synthesis:

(163) ##STR00043##

Step 1: Synthesis of Chloromethyl Propyl Carbonate (Compound 12)

(164) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and n-propanol (1.41 g, 23.45 mmol), which were dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 2.86 g in a yield of 80.2%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(propylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-7)

(165) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 12 (630 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol), and 10 ml DMF. The reaction was heated to 60° C. and reacted overnight, then cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 127 mg of product in a yield of 21.3%. LC-MS(APCI): m/z=720.5 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.33 (s, 1H), 8.02 (s, 1H), 6.06 (s, 2H), 5.61 (dd, J=28.2, 8.9 Hz, 2H), 4.91 (m, 1H), 4.40-4.12 (m, 4H), 3.94 (m, 3H), 3.68 (t, J=10.0 Hz, 1H), 2.89 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.69 (m 2H), 1.64 (s, 2H), 1.33-1.22 (m, 32H), 0.86 (t, J=6.6 Hz, 3H).

Example 10

(R)-9-{2-[(hexadecyldisulfanylethyl)(n-butylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-8)

(166) ##STR00044##

(167) The following synthetic route was used for the synthesis:

(168) ##STR00045##

Step 1: Synthesis of Chloromethyl Butyl Carbonate (Compound 13)

(169) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and n-butanol (1.74 g, 23.45 mmol), which were dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 2.94 g in a yield of 75.6%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(butylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-8)

(170) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 13 (688 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 119 mg of product in a yield of 19.6%. LC-MS(APCI): m/z=734.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.33 (s, 1H), 8.02 (s, 1H), 6.06 (s, 2H), 5.61 (dd, J=28.2, 8.9 Hz, 2H), 4.91 (m, 1), 4.40-4.12 (m, 4H), 3.94 (m, 3H), 3.68 (t, J=10.0 Hz, 1H), 2.89 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.69 (m 2H), 1.64 (s, 2H), 1.33-1.22 (m, 34H), 0.86 (t, J=6.6 Hz, 3H).

Example 11

(R)-9-{2-[(hexadecyldisulfanylethyl)(isobutylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-9)

(171) ##STR00046##

(172) The following synthetic route was used for the synthesis:

(173) ##STR00047##

Step 1: Synthesis of Isobutyl Chloromethyl Carbonate (Compound 14)

(174) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and isobutanol (1.74 g, 23.45 mmol), which were dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added the mixture dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 3.06 g in a yield of 78.7%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(isobutylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-9)

(175) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 14 (688 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 134 mg of product in a yield of 22.0%. LC-MS(APCI): m/z=734.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (dd, J=28.2, 8.9 Hz, 2H), 4.91 (m, 1H), 4.40-4.12 (m, 4H), 3.94 (m, 3H), 3.68 (t, J=10.0 Hz, 1H), 2.89 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.69 (m 1H), 1.64 (s, 2H), 1.33-1.22 (m, 35H), 0.86 (t, J=6.6 Hz, 3H).

Example 12

(R)-9-{2-[(hexadecyldisulfanylethyl)(s-butylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-10)

(176) ##STR00048##

(177) The following synthetic route was used for the synthesis:

(178) ##STR00049##

Step 1: Synthesis of Sec-Butyl Chloromethyl Carbonate (Compound 15)

(179) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and sec-butanol (1.74 g, 23.45 mmol), which was dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added the mixture dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 2.99 g in a yield of 76.8%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(s-butylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-10)

(180) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 15 (688 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 112 mg of product in a yield of 18.4%. LC-MS(APCI): m/z=734.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.31 (s, 1H), 8.01 (s, 1H), 6.06 (s, 2H), 5.62 (m, 2H), 4.93 (m, 1H), 4.40-4.12 (m, 4H), 3.94 (m, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.92 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.69 (m 1H), 1.64 (s, 2H), 1.33-1.22 (m, 36H), 0.88 (t, J=6.6 Hz, 3H).

Example 13

(R)-9-{2-[(hexadecyldisulfanylethyl)(t-butylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-11)

(181) ##STR00050##

(182) The following synthetic route was used for the synthesis:

(183) ##STR00051##

Step 1: Synthesis of Tert-Butyl Chloromethyl Carbonate (Compound 16)

(184) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and tert-butanol (1.74 g, 23.45 mmol), which were dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 3.25 g in a yield of 83.5%.

(185) Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(t-butylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-11)

(186) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 16 (688 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 134 mg of product in a yield of 22.0%. LC-MS(APCI): m/z=734.2 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (m, 2H), 4.92 (dd, J=13.2, 6.6 Hz, 1H), 4.40-4.12 (m, 3H), 3.92 (dd, J=11.5, 7.1 Hz, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.87 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.64 (s, 2H), 1.33-1.22 (m, 38H), 0.87 (t, J=6.6 Hz, 3H).

Example 14

(R)-9-{2-[(hexadecyldisulfanylethyl)(neopentylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-12)

(187) ##STR00052##

(188) The following synthetic route was used for the synthesis:

(189) ##STR00053##

Step 1: Synthesis of Neopentyl Chloromethyl Carbonate (Compound 17)

(190) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and neopentyl alcohol (2.07 g, 23.45 mmol), which were dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 3.58 g in a yield of 84.8%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(neopentylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-12)

(191) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 17 (746 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 98 mg of product in a yield of 15.8%. LC-MS(APCI): m/z=748.6 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (m, 2H), 4.92 (dd, J=13.2, 6.6 Hz, 1H), 4.40-4.12 (m, 4H), 3.92 (dd, J=11.5, 7.1 Hz, 3H), 3.68 (t, J=10.0 Hz, 1H), 2.87 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.64 (s, 2H), 1.33-1.22 (m, 38H), 0.87 (t, J=6.6 Hz, 3H).

Example 15

(R)-9-{2-[(hexadecyldisulfanylethyl)(cyclopentylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-13)

(192) ##STR00054##

(193) The following synthetic route was used for the synthesis:

(194) ##STR00055##

Step 1: Synthesis of Cyclopentyl Chloromethyl Carbonate (Compound 18)

(195) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and cyclopentanol (2.02 g, 23.45 mmol), which were dissolved with 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The reaction was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, which was dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 3.27 g in a yield of 78.4%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(cyclopentylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-13)

(196) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 18 (737.8 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 114 mg of product in a yield of 18.4%. LC-MS(APCI): m/z=746.8 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (m, 2H), 4.92 (dd, J=13.2, 6.6 Hz, 1H), 4.40-4.12 (m, 4H), 3.92 (dd, J=11.5, 7.1 Hz, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.87 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.64-1.57 (m, 6H), 1.33-1.22 (m, 33H), 0.87 (t, J=6.6 Hz, 3H).

Example 16

(R)-9-{2-[(hexadecyldisulfanylethyl)(cyclohexylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-14)

(197) ##STR00056##

(198) The following synthetic route was used for the synthesis:

(199) ##STR00057##

Step 1: Synthesis of Cyclohexyl Chloromethyl Carbonate (Compound 19)

(200) To a reaction flask were added chloromethyl chloroformate (3.0 g, 23.45 mmol) and cyclohexanol (2.35 g, 23.45 mmol), which was dissolved in 20 ml anhydrous ether. The reaction was cooled to 0° C., and pyridine (1.85 g, 23.45 mmol) was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and stirred overnight. The mixture was diluted with water, washed three times with 1% citric acid, saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to afford the product 3.47 g in a yield of 77.1%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylethyl)(cyclohexylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-14)

(201) To a reaction flask were added Compound 7 (500 mg, 0.83 mmol), Compound 19 (796 mg, 4.14 mmol), potassium carbonate (572.7 mg, 4.14 mmol) and potassium iodide (68.1 mg, 0.41 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight. The reaction was cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 147 mg of product in a yield of 23.3%. LC-MS(APCI): m/z=760.9 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (m, 2H), 4.92 (dd, J=13.2, 6.6 Hz, 1H), 4.40-4.12 (m, 4H), 3.92 (dd, J=11.5, 7.1 Hz, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.87 (m, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.66-1.58 (m, 6H), 1.33-1.22 (m, 35H), 0.87 (t, J=6.6 Hz, 3H).

Example 17

(2′R,1′S)-9-{2-[(dodecyldisulfanylethyl)(isopropoxycarbonyl-1-ylethylamino)phosphorylmethoxy]propyl}adenine (Compound T-15)

(202) ##STR00058##

(203) The following synthetic route was used for the synthesis:

(204) ##STR00059##

Step 1: Synthesis of L-Isopropyl Alanine (Compound 20)

(205) To a reaction flask was added L-isopropyl alanine hydrochloride (10.0 g, 59.9 mmol), which was dissolved in 40 ml anhydrous dichloromethane. Sodium bicarbonate (15.0 g, 149.7 mmol) was added to the mixture, and gas was released. The reaction was stirred at room temperature overnight. Insoluble material was filtered off. The organic phase was washed 3 times with saturated brine, dried over anhydrous sodium sulfate, and concentrated to afford the product 7.54 g in a yield of 96.0%.

Step 2: Synthesis of (2′R,1′S)-9-{2-[(dodecyldisulfanylethyl)(isopropoxycarbonyl-1-ylethylamino)phosphorylmethoxy]propyl}adenine (Compound T-15)

(206) To a reaction flask were added Compound 3 (1.5 g, 2.74 mmol) and anhydrous DMF (240 mg, 3.29 mmol), which were dissolved in 30 ml anhydrous dichloromethane. Oxalyl chloride (6.85 ml, 13.7 mmol) was slowly added to the mixture dropwise at room temperature. After the addition, the reaction was stirred for 2-3 hours under the nitrogen protection. The reaction was concentrated to remove the solvent and excess oxalyl chloride. The residue was dissolved in 20 ml anhydrous dichloromethane, which was cooled to 0° C. Under the nitrogen protection, a solution of Compound 20 (1.8 g, 13.7 mmol) and triethylamine (1.39 g, 13.7 mmol) in dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and reacted for 1 hour. Upon TLC detection showed the reaction was completed, 2 ml ethanolamine was added to the mixture and the mixture was stirred overnight. The mixture was diluted with a small amount of water. The organic phase was separated, washed 3 times with saturated brine, concentrated, and purified by silica gel column chromatography to afford 315 mg of product in a yield of 17.4%. LC-MS(APCI): m/z=661.1 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (d, J=4.3 Hz, 1H), 8.01 (d, J=14.2 Hz, 1H), 5.87 (s, 2H), 4.97 (ddt, J=43.9, 12.5, 6.2 Hz, 1H), 4.44-4.30 (m, 1H), 4.27-4.08 (m, 3H), 3.96 (ddd, J=19.1, 10.8, 3.5 Hz, 2H), 3.87-3.76 (m, 1H), 3.56 (ddd, J=13.1, 10.1, 6.9 Hz, 1H), 2.87 (ddd, J=19.6, 10.9, 6.5 Hz, 2H), 2.68 (td, J=7.3, 5.3 Hz, 2H), 1.69-1.60 (m, 2H), 1.41-1.31 (m, 6H), 1.30-1.20 (m, 24H), 0.87 (t, J=6.8 Hz, 3H).

Example 18

(2′R,1′S)-9-{2-[(tetradecyldisulfanylethyl)(isopropoxycarbonyl-1-ylethylamino)phosphorylmethoxy]propyl}adenine (Compound T-16)

(207) ##STR00060##

(208) The following synthetic route was used for the synthesis:

(209) ##STR00061##

(210) To a reaction flask were added Compound 5 (979 mg, 1.7 mmol) and anhydrous DMF (150 mg, 2.04 mmol), which was dissolved in 20 ml anhydrous dichloromethane. Oxalyl chloride (4.25 ml, 8.51 mmol) was slowly added to the mixture dropwise at room temperature. After the addition, the reaction was stirred for 2-3 hours under the nitrogen protection. The mixture was concentrated to remove the solvent and excess oxalyl chloride. The residue was dissolved in 20 ml anhydrous dichloromethane, and the reaction was cooled to 0° C. Under the nitrogen protection, a solution of Compound 20 (1.11 g, 8.5 mmol) and triethylamine (861.1 mg, 8.5 mmol) in dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and reacted for 1 hour. Upon TLC detection showed the reaction was completed, 2 ml ethanolamine was added to the mixture and the mixture was stirred overnight. The mixture was diluted with a small amount of water. The organic phase was separated, washed 3 times with saturated brine, concentrated, and purified by silica gel column chromatography to afford 146 mg of product in a yield of 12.5%. LC-MS(APCI): m/z=689.5 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (d, J=4.3 Hz, 1H), 8.00 (d, J=13.1 Hz, 1H), 5.79 (s, 2H), 4.98 (dtd, J=44.5, 12.5, 6.2 Hz, 1H), 4.38 (ddd, J=19.6, 14.4, 3.0 Hz, 1H), 4.26-4.06 (m, 3H), 3.96 (tdd, J=18.1, 10.3, 3.3 Hz, 2H), 3.87-3.75 (m, 1H), 3.61-3.51 (m, 1H), 2.92-2.80 (m, 2H), 2.67 (dt, J=12.4, 7.2 Hz, 2H), 1.65 (dd, J=13.1, 7.0 Hz, 2H), 1.38-1.20 (m, 34H), 0.87 (t, J=6.8 Hz, 3H).

Example 19

(2′R,1′S)-9-{2-[(hexadecyldisulfanylethyl)(isopropoxycarbonyl-1-ylethylamino)phosphorylmethoxy]propyl}adenine (Compound T-17)

(211) ##STR00062##

(212) The following synthetic route was used for the synthesis:

(213) ##STR00063##

(214) To a reaction flask were added Compound 7 (800 mg, 1.33 mmol) and anhydrous DMF (194.5 mg, 2.66 mmol), which were dissolved in 15 ml anhydrous dichloromethane. Oxalyl chloride (3.32 ml, 6.63 mmol) was added dropwise to the mixture slowly at room temperature. After the addition, the reaction was stirred for 2-3 hours under the nitrogen protection. The reaction was concentrated to remove the solvent and excess oxalyl chloride, which were dissolved in 15 ml anhydrous dichloromethane. The reaction was cooled to 0° C., and a solution of Compound 20 (698 mg, 5.32 mmol) and triethylamine (671 mg, 6.63 mmol) in dichloromethane was slowly added to the mixture dropwise under the nitrogen protection. After the addition, the reaction was warmed to rt and reacted for 1 hour. Upon TLC detection showed the reaction was completed, 2 ml ethanolamine was added to the mixture and the mixture was stirred overnight. The mixture was diluted with a small amount of water. The organic phase was separated, washed 3 times with saturated brine, concentrated, and purified by silica gel column chromatography to afford 127 mg of product in a yield of 13.3%. LC-MS(APCI): m/z=717.4 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (d, J=4.4 Hz, 1H), 7.99 (d, J=12.2 Hz, 1H), 5.76 (d, J=7.0 Hz, 2H), 4.97 (ddt, J=44.7, 12.5, 6.3 Hz, 1H), 4.38 (ddd, J=19.6, 14.4, 3.0 Hz, 1H), 4.26-4.07 (m, 3H), 4.02-3.90 (m, 2H), 3.86-3.76 (m, 1H), 3.56 (ddd, J=13.2, 10.1, 7.6 Hz, 1H), 2.87 (ddd, J=20.3, 10.2, 5.7 Hz, 2H), 2.72-2.62 (m, 2H), 1.69-1.59 (m, 2H), 1.35 (dd, J=9.3, 7.2 Hz, 6H), 1.29-1.20 (m, 32H), 0.87 (t, J=6.8 Hz, 3H).

Example 20

(2′R,1′S)-9-{2-[(octadecyldisulfanylethyl)(isopropoxycarbonyl-1-ylethylamino)phosphorylmethoxy]propyl}adenine (Compound T-18)

(215) ##STR00064##

(216) The following synthetic route was used for the synthesis:

(217) ##STR00065##

(218) To a reaction flask were added Compound 9 (800 mg, 1.27 mmol) and anhydrous DMF (111 mg, 1.52 mmol), which were dissolved in 15 ml anhydrous dichloromethane. Oxalyl chloride (3.17 ml, 6.33 mmol) was slowly added to the mixture dropwise at room temperature. After the addition, the reaction was stirred for 2-3 hours under the nitrogen protection. The reaction was concentrated to remove the solvent and excess oxalyl chloride. The residue was dissolved in 15 ml anhydrous dichloromethane, and the reaction was cooled to 0° C. Under the nitrogen protection, a solution of Compound 20 (665 mg, 5.07 mmol) and triethylamine (640 mg, 6.33 mmol) in dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and reacted for 1 hour. Upon TLC detection showed the reaction was completed, 2 ml ethanolamine was added to the mixture and the mixture was stirred overnight. The mixture was diluted with a small amount of water. The organic phase was separated, washed 3 times with saturated brine, concentrated, and purified by silica gel column chromatography to afford 136 mg of product in a yield of 13.3%. LC-MS(APCI): m/z=745.7 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (d, J=4.3 Hz, 1H), 7.99 (d, J=12.8 Hz, 1H), 5.79 (d, J=6.0 Hz, 2H), 4.97 (ddt, J=44.0, 12.5, 6.3 Hz, 1H), 4.44-4.31 (m, 1H), 4.17 (dddd, J=23.9, 22.1, 10.8, 6.5 Hz, 3H), 3.96 (tdd, J=18.5, 10.5, 3.6 Hz, 2H), 3.86-3.74 (m, 1H), 3.56 (ddd, J=13.1, 10.1, 6.9 Hz, 1H), 2.93-2.80 (m, 2H), 2.68 (td, J=7.3, 5.4 Hz, 2H), 1.65 (dq, J=14.4, 7.0 Hz, 2H), 1.35 (dd, J=8.8, 7.3 Hz, 6H), 1.30-1.20 (m, 36H), 0.87 (t, J=6.8 Hz, 3H).

Example 21

(R)-9-{2-[(hexadecyldisulfanylpropyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-19)

(219) ##STR00066##

(220) The following synthetic route was used for the synthesis:

(221) ##STR00067##

Step 1: Synthesis of 3-(hexadecyldisulfanyl)propanol (Compound 21)

(222) To a reaction flask were added 3-mercaptopropanol (2.76 g, 30 mmol), and n-hexadecanethiol (7.75 g, 30 mmol), which were dissolved in 105 ml anhydrous methanol and 105 ml anhydrous dichloromethane. Pyridine (4.74 g, 60 mmol) was added to the mixture. Under the nitrogen protection, elemental iodine (7.61 g, 30 mmol) was added to the mixture in portions. After the addition, the reaction was stirred at room temperature for 2-5 hours. Insoluble material was filtered off upon TLC detection showed the reaction was completed, and the filtrate was concentrated to dryness. The residue was dissolved in a small amount of dichloromethane, washed with saturated brine, concentrated, purified by column chromatography, and dried in vacuo to afford product 3.42 g in a yield of 32.7%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylpropyl)phosphorylmethoxy]propyl}adenine (Compound 22)

(223) To a reaction flask was added Compound 2 (2.43 g, 6.44 mmol), which was dissolved in 20 ml anhydrous dichloromethane. Under the nitrogen protection, the reaction was cooled to 0° C., and a solution of Compound 21 (2.69 g, 7.73 mmol) and pyridine (3.05 g, 38.64 mmol) in anhydrous dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was reacted at the low temperature for 10 minutes and warmed to rt and reacted for 2 hours. Upon MS monitoring showed the reaction was completed, 1 M dilute hydrochloric acid (30 ml) was added to quench the reaction, and the reaction was stirred at room temperature overnight. The organic phase was separated upon MS monitoring showed the reaction was completed. The aqueous phase was extracted 2-3 times with dichloromethane. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 2.67 g of product in a yield of 67.2%. LC-MS(APCI): m/z=618.5 (M+1).sup.+.

Step 3: Synthesis of (R)-9-{2-[(hexadecyldisulfanylpropyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-19)

(224) To a reaction flask were added Compound 22 (1.0 g, 1.62 mmol), chloromethyl isopropyl carbonate (1.23 g, 8.1 mmol), potassium carbonate (1.12 g, 8.1 mmol) and potassium iodide (134.5 mg, 0.81 mmol). 15 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight. The reaction was cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 163 mg of product in a yield of 13.7%. LC-MS(APCI): m/z=734.1 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (m, 2H), 4.92 (dd, J=13.2, 6.6 Hz, 1H), 4.40-4.12 (m, 4H), 3.92 (dd, J=11.5, 7.1 Hz, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.87 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.82-1.64 (m, 4H), 1.33-1.22 (m, 35H), 0.87 (t, J=6.6 Hz, 3H).

Example 22

(2′R,1′S)-9-{2-[(hexadecyldisulfanylpropyl)(isopropoxycarbonyl-1-ylethylamino)phosphorylmethoxy]propyl}adenine (Compound T-20)

(225) ##STR00068##

(226) The following synthetic route was used for the synthesis:

(227) ##STR00069##

(228) To a reaction flask were added Compound 22 (1.02 g, 1.66 mmol) and anhydrous DMF (145.6 mg, 1.99 mmol), which were dissolved in 20 ml anhydrous dichloromethane. Oxalyl chloride (4.14 ml, 8.28 mmol) was slowly added to the mixture dropwise at room temperature. After the addition, the reaction was stirred for 2-3 hours under the nitrogen protection. The reaction was concentrated to remove the solvent and excess oxalyl chloride, which were dissolved in 20 ml anhydrous dichloromethane. The reaction was cooled to 0° C. Under the nitrogen protection, a solution of Compound 20 (871 mg, 6.64 mmol) and triethylamine (837.8 mg, 8.28 mmol) in dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was warmed to rt and reacted for 1 hour. Upon TLC detection showed the reaction was completed, 3 ml ethanolamine was added to the mixture, which was stirred overnight. The mixture was diluted with a small amount of water. The organic phase was separated, washed 3 times with saturated brine, concentrated, and purified by silica gel column chromatography to afford 167 mg of product in a yield of 13.8%. LC-MS(APCI): m/z=731.7 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (d, J=4.4 Hz, 1H), 7.99 (d, J=12.2 Hz, 1H), 5.76 (d, J=7.0 Hz, 2H), 4.97 (ddt, J=44.7, 12.5, 6.3 Hz, 1H), 4.38 (ddd, J=19.6, 14.4, 3.0 Hz, 1H), 4.26-4.07 (m, 3H), 4.02-3.90 (m, 2H), 3.86-3.76 (m, 1H), 3.56 (ddd, J=13.2, 10.1, 7.6 Hz, 1H), 2.87 (ddd, J=20.3, 10.2, 5.7 Hz, 2H), 2.72-2.62 (m, 2H), 1.69-1.59 (m, 4H), 1.35 (dd, J=9.3, 7.2 Hz, 6H), 1.29-1.20 (m, 32H), 0.87 (t, J=6.8 Hz, 3H).

Example 23

(R)-9-{2-[(hexadecyldisulfanylbutyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-21)

(229) ##STR00070##

(230) The following synthetic route was used for the synthesis:

(231) ##STR00071##

Step 1: Synthesis of 4-(hexadecyldisulfanyl)butanol (Compound 23)

(232) To a reaction flask were added 4-mercaptobutanol (2.12 g, 20 mmol), and n-hexadecanethiol (5.16 g, 20 mmol), which were dissolved in 70 ml anhydrous methanol and 70 ml anhydrous dichloromethane. Pyridine (3.16 g, 40 mmol) was added. Under the nitrogen protection, elemental iodine (5.08 g, 20 mmol) was added to the mixture in portions. After the addition, the reaction was stirred at room temperature for 2-5 hours. Insoluble material was filtered off upon TLC detection showed the reaction was completed, and the filtrate was concentrated to dryness. The residue was dissolved in a small amount of dichloromethane, washed with saturated brine, concentrated, purified by column chromatography, and dried in vacuo to afford product 2.8 g in a yield of 38.7%.

Step 2: Synthesis of (R)-9-{2-[(hexadecyldisulfanylbutyl)phosphorylmethoxy]propyl}adenine (Compound 24)

(233) To a reaction flask was added Compound 2 (2.43 g, 6.44 mmol), which was dissolved in 20 ml anhydrous dichloromethane. Under the nitrogen protection, the reaction was cooled to 0° C., and a solution of Compound 23 (2.8 g, 7.73 mmol) and pyridine (3.05 g, 38.64 mmol) in anhydrous dichloromethane was slowly added to the mixture dropwise. After the addition, the reaction was reacted at the low temperature for 10 minutes and warmed to rt and reacted for 2 hours. Upon MS monitoring showed the reaction was completed, 1 M dilute hydrochloric acid (30 ml) was added to quench the reaction, and the mixture was stirred at room temperature overnight. Upon MS monitoring showed the reaction was completed, the organic phase was separated. The aqueous phase was extracted 2-3 times with dichloromethane. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 2.99 g of product in a yield of 73.6%. LC-MS(APCI): m/z=632.6 (M+1).sup.+.

Step 3: Synthesis of (R)-9-{2-[(hexadecyldisulfanylbutyl)(isopropylcarbonatemethyl)phosphorylmethoxy]propyl}adenine (Compound T-21)

(234) To a reaction flask were added Compound 24 (500 mg, 0.79 mmol), chloromethyl isopropyl carbonate (602 mg, 3.96 mmol), potassium carbonate (547.3 mg, 3.96 mmol) and potassium iodide (65.7 mg, 0.4 mmol). 10 ml DMF was added to the mixture. The reaction was heated to 60° C. and reacted overnight, and cooled to room temperature upon TLC detection showed the reaction was completed. The mixture was diluted with excess amount of water, and extracted 3-4 times with ethyl acetate. The organic phase was combined, washed with saturated brine, concentrated, and purified by silica gel column chromatography to afford 103 mg of product in a yield of 17.4%. LC-MS(APCI): m/z=748.5 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 8.03 (s, 1H), 6.06 (s, 2H), 5.61 (m, 2H), 4.92 (dd, J=13.2, 6.6 Hz, 1H), 4.40-4.12 (m, 4H), 3.92 (dd, J=11.5, 7.1 Hz, 2H), 3.68 (t, J=10.0 Hz, 1H), 2.87 (dt, J=19.2, 6.7 Hz, 2H), 2.67 (dd, J=11.8, 7.0 Hz, 2H), 1.75-1.62 (m, 6H), 1.33-1.22 (m, 35H), 0.87 (t, J=6.6 Hz, 3H).

Example 24

(2′R,1′S)-9-{2-[(hexadecyldisulfanylbutyl)(isopropoxycarbonyl-1-ylethylamino)phosphorylmethoxy]propyl}adenine (Compound T-22)

(235) ##STR00072##

(236) The following synthetic route was used for the synthesis:

(237) ##STR00073##

(238) To a reaction flask were added Compound 24 (700 mg, 1.11 mmol) and anhydrous DMF (97.4 mg, 1.33 mmol), which were dissolved in 15 ml anhydrous dichloromethane. Oxalyl chloride (2.77 ml, 5.54 mmol) was slowly added to the mixture dropwise at room temperature. After the addition, the reaction was stirred for 2-3 hours under the nitrogen protection, and concentrated to remove the solvent and excess oxalyl chloride. The residue was dissolved in 15 ml anhydrous dichloromethane. The reaction was cooled to 0° C., and a solution of Compound 20 (726.7 mg, 5.54 mmol) and triethylamine (560.6 mg, 5.54 mmol) in dichloromethane was slowly added to the mixture dropwise under the nitrogen protection. After the addition, the reaction was warmed to rt and reacted for 1 hour. Upon TLC detection showed the reaction was completed, 2 ml ethanolamine was added to the mixture, which was stirred overnight. The mixture was diluted with a small amount of water. The organic phase was separated, washed 3 times with saturated brine, concentrated, and purified by silica gel column chromatography to afford 149 mg of product in a yield of 18.0%. LC-MS(APCI): m/z=745.1 (M+1).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (d, J=4.4 Hz, 1H), 7.99 (d, J=12.2 Hz, 1H), 5.76 (d, J=7.0 Hz, 2H), 4.97 (ddt, J=44.7, 12.5, 6.3 Hz, 1H), 4.38 (ddd, J=19.6, 14.4, 3.0 Hz, 1H), 4.26-4.07 (m, 3H), 4.02-3.90 (m, 2H), 3.86-3.76 (m, 1H), 3.56 (ddd, J=13.2, 10.1, 7.6 Hz, 1H), 2.87 (ddd, J=20.3, 10.2, 5.7 Hz, 2H), 2.72-2.62 (m, 2H), 1.72-1.55 (m, 6H), 1.35 (dd, J=9.3, 7.2 Hz, 6H), 1.29-1.20 (m, 32H), 0.87 (t, J=6.8 Hz, 3H).

(239) Biological Activity Test

(240) (1) Detection of In Vitro Anti-HIV Activity of Compounds

(241) The data was analyzed by GraphPad Prism software, and the curve was fitted and EC.sub.50 and CC.sub.50 values were calculated. EC.sub.50 refers to the effective concentration that inhibits 50% virus production, 50% virus infectivity, or 50% virus-induced cellular effects. CC.sub.50 refers to the inhibitory concentration that reduces the cell growth or viability of uninfected cells by 50%.

(242) Compound treatment: the test compounds and the reference compounds were 2-fold diluted with DMSO and added to the cell culture plate. The test compounds and the reference compounds were tested at 8 concentrations in duplicate.

(243) Viral infection and cell treatment: HIV-1 and MT-4 cells were co-incubated at 37° C., in an incubator with 5% CO.sub.2 for 1 h. The infected cells were then seeded in a cell culture plate at a certain density. The final concentration of DMSO in the cell culture medium was 0.5%. The cells were incubated at 37° C., in an incubator with 5% CO.sub.2 for 5 days. The cells tested in the cytotoxicity assay were uninfected MT-4 cells, and other test conditions were with the same as that of the antiviral activity assay.

(244) The activity assay of the cell: the cell viability was tested by using cell viability assay reagent CellTiter-glo (Promega). The raw data was used for calculating the anti-HIV-1 activity and cytotoxicity of the Compounds. Dose-response curves of compounds and their EC.sub.50 and CC.sub.50 values were obtained by analysis with GraphPad Prism software, where A represents EC.sub.50≤5 nM, B represents 5 nM<EC.sub.50≤20 nM, C represents 20 nM<EC.sub.50≤50 nM; D represents 100 nM<CC.sub.50≤500 nM, E represents 500 nM<CC.sub.50≤1000 nM, F represents 1000 nM<CC.sub.50≤5000 nM, and G represents CC.sub.50>5000 nM (as shown in Table 1 below).

(245) (2) Detection of Anti-HBV Activity of Compounds In Vitro

(246) Experimental method: anti-HBV activity of compounds was detected by Bright-Glo (Promega) luciferase. The data was analyzed by GraphPad Prism software, and the curve was fitted and EC.sub.50 and CC.sub.50 values were calculated. EC.sub.50 refers to the effective concentration that inhibits 50% virus production, 50% virus infectivity, or 50% virus-induced cellular effects. CC.sub.50 refers to the inhibitory concentration that reduces the cell growth or viability of uninfected cells by 50%.

Experimental Steps

(247) Anti-cell viability assay: in vitro anti-hepatitis b virus activity of Example compounds was tested in HepG2.2.15 cells with TDF as a positive control compound. Cells were seeded into a 96-well plate on the first day; the compounds were added to treat the cells on the next day; and a new culture medium containing the compounds was replaced on the fifth day. On the eighth day, the supernatant was collected for DNA extraction. The content of HBV DNA was detected by quantitative PCR. Both test compound and TDF were subjected to a 3-fold serial dilution to get 8 concentration points in duplicate. The final concentration of DMSO in the culture medium was 0.5%. The formula for calculating the percentage of inhibition is as follows:
Inhibition rate=(1−copy number of HBV in sample/copy number of HBV in DMSO control group)×100%

(248) EC.sub.50 is analyzed by Graphpad Prism software (four parameter logistic equations), wherein I represents EC.sub.50≤15 nM, II represents 15 nM≤EC.sub.50≤50 nM, III represents 50 nM<EC.sub.50≤100 nM, IV represents 100 nM<EC.sub.50≤400 nM (as shown in Table 1 below).

(249) Cytotoxicity experiments: the arrangement of compounds on plate and compound-treatment process were the same as that of anti-HIV activity. Six days after the cells were treated by compounds, the cell activity was tested. Cell-titer Blue reagent was added to each well, the cells were incubated at 37° C. for 3 hours, and fluorescence values were read (560Ex/590 Em). The data were analyzed and the relative cell viability was calculated:

(250) The percentage of cell viability was calculated using the following formula:
% cell viability=(fluorescence reading of sample−fluorescence reading of culture control)/(fluorescence reading of DMSO control−fluorescence reading of culture control)×100.

(251) Finally, the CC.sub.50 value of the compounds were calculated by the GraphPad Prism software. V represents 10000 nM<CC.sub.50≤50000 nM, VI represents CC.sub.50>50000 nM (as shown in Table 1 below).

(252) TABLE-US-00001 TABLE 1 HBV activity and HIV activity of example compounds HIV activity and HBV activity and Compound cytotoxicity data cytotoxicity data No. EC.sub.50 (nM) CC.sub.50 (nM) EC.sub.50 (nM) CC.sub.50 (nM) T-1 C F IV VI T-2 A F III VI T-3 A F I VI T-3-1 A D IV VI T-3-2 A D IV VI T-4 B G II VI T-5 A F III VI T-6 A E III VI T-7 A D II VI T-8 B E II VI T-9 A E II VI T-10 A D III VI T-11 A F I VI T-12 A F III VI T-13 A F II VI T-14 A E III VI T-15 B F I V T-16 B F I VI T-17 B E I VI T-18 C F IV VI T-19 A G IV VI T-20 B F II V T-21 C G IV VI T-22 B G I V

(253) Experimental results show that the compounds disclosed herein have strong anti-HIV activity and anti-HBV activity (both in nanomolar levels). In addition, in the cell lines tested, the compounds disclosed herein do not show toxicity (the preferable CC.sub.50>50000 nM).

(254) (3) Liver Microsomal Metabolism Experiment

(255) Microsomal experiments: human liver microsomes: 0.5 mg/mL, Xenotech; rat liver microsomes: 0.5 mg/mL, Xenotech; coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

(256) Preparation of stock solution: powder of Example compounds were precisely weighed and dissolved in DMSO to 5 mM.

(257) Preparation of phosphate buffer (100 mm, pH 7.4): 0.5M pre-formulated potassium dihydrogen phosphate (150 ml) and 0.5M potassium hydrogen phosphate (700 ml) were mixed well. The pH of the mixture was then adjusted to 7.4 with 0.5M potassium hydrogen phosphate solution. The resulting solution was 5-fold diluted with ultrapure water before use, and magnesium chloride was added to the solution, affording a phosphate buffer (100 mm), containing potassium phosphate (100 mM), magnesium chloride (3.3 mM), pH 7.4.

(258) A NADPH regeneration system solution (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was formulated, which was placed on wet ice before use.

(259) Preparation of stop solution: 50 ng/mL of propranolol hydrochloride and 200 ng/mL of tolbutamide (internal standard) in acetonitrile. 25057.5 μL of phosphate buffer (pH 7.4) was placed to a 50 mL centrifuge tube, and 812.5 μL human liver microsomes were added to the tube, which were mixed well to give a protein concentration of 0.625 mg/mL liver microsomes diluent. 25057.5 μL of phosphate buffer (pH 7.4) was placed to a 50 ml centrifuge tube, and 812.5 μL SD rat liver microsomes were added to the tube, which were mixed well to give a protein concentration of 0.625 mg/mL liver microsomes diluent.

(260) Incubation of the sample: the stock solution of the corresponding compounds was diluted to 0.25 mM with 70% acetonitrile in water as a working solution for later use. 398 μL of human liver microsomes dilution or rat liver microsomes dilution was added into 96-well incubation plate (N=2), and 2 μL of 0.25 mM working solution was added to the plate, which were mixed well.

(261) Metabolic stability assay: 300 μL of pre-chilled stop solution was added to each well of a 96-well deep-well plate and the plate was placed on ice as a stop plate. The 96-well incubation plate and NADPH regeneration system were placed in a 37° C. water bath, shaked at 100 rpm, pre-incubated for 5 min. 80 μL of incubation solution was taken from each well of the incubation plate and was added to the stop plate, which were mixed well. 20 μL of NADPH regeneration system solution was supplemented into the mixture to form the 0 min sample. Then 80 μL of NADPH regeneration system solution was added to each well of the incubation plate to initiate the reaction and start timing. The reaction concentration of the corresponding compound was 1 μM, and the protein concentration was 0.5 mg/mL. At 10, 30, and 90 min, 100 μL of the reaction solution was added to the stop plate and vortexed for 3 min to quench the reaction. The stop plate was centrifuged at 5000×g at 4° C. for 10 min. 100 μL of supernatant was taken into a 96-well plate containing 100 μL of distilled water, which were mixed well. LC-MS/MS was used for sample analysis.

(262) Data analysis: The peak area of the corresponding compounds and internal standard was detected by LC-MS/MS system, and the peak area ratio of compounds to internal standard was calculated. The slope was measured by plotting the natural log of the percentage of the remaining amount of the compounds versus time, and t.sub.1/2 and CL.sub.int were calculated according to the following formula, where V/M is equal to 1/protein concentration.

(263) $t_{1/2}=-\frac{0.693}{\mathrm{slope}},{\mathrm{CL}}_{\mathrm{int}}=\frac{0.693}{t_{1/2}}.Math.\frac{V}{M}$

(264) The compounds disclosed herein were tested in the above microsome experiment and it was found that the compounds disclosed herein have superior metabolic stability. The results of the human liver microsome experiment and the rat liver microsome experiment of representative examples are summarized in Table 2 below.

(265) TABLE-US-00002 TABLE 2 Evaluation of liver microsome metabolism of Example compounds Human liver Rat liver microsome experiment microsome experiment No. t.sub.1/2(min) CL.sub.int (μL/min/mg) t.sub.1/2(min) CL.sub.int (μL/min/mg) T-2 37.3 37.2 43.6 31.8 T-3 79.8 17.4 37.4 37.0 T-3-2 19.5 71.0 25.5 54.4 T-4 192.2 7.2 T-5 19.1 72.6 16.8 82.3 T-6 22.3 62.1 24.5 56.5 T-7 26.8 51.7 49.3 28.1 T-8 67.5 20.5 48.2 28.8 T-9 124.2 11.2 103.9 13.3 T-10 58.3 23.8 55.2 25.1 T-11 175.4 7.9 78.9 17.6 T-12 216.2 6.4 179.1 7.7 T-13 108.0 12.8 77.8 17.8 T-14 1509 0.92 241.1 5.7 T-15 9.2 149.9 8.4 164.5 T-16 11.5 120.6 12.5 110.8 T-17 54.4 25.5 69.6 19.9 T-18 627.6 2.2 1117 1.24 T-19 59.1 23.5 57.5 24.1 T-20 115.9 12.0 128.3 10.8 T-21 63.7 21.7 77.9 17.8 T-22 225.7 6.1 636.9 2.2

(266) The experimental results are shown in Table 2 above. The compounds disclosed herein have a longer half-life and a lower clearance rate. They show superior metabolic stability in human liver microsomes and rat liver microsomes, and are more suitable as drugs for anti-HIV and/or anti-HBV infection

(267) (4) Rat Pharmacokinetics

(268) Experimental purpose: To study the pharmacokinetic behavior of the compounds disclosed herein after they were administered to the rats.

(269) Experimental Animals:

(270) Species and strain: SD rats, grade: SPF grade

(271) Gender and number: male, 6

(272) Weight range: 180-220 g (actual body weight in the range of 187-197 g)

(273) Source: Shanghai Sippr-BK laboratory animal Co. Ltd.

(274) Assays and animal certification number: SCXK(Shanghai)2013-0016

(275) Experiment Procedure: Before the blood sample collection, 20 μL of 2M sodium fluoride solution (esterase inhibitor) was added to the EDTA-K2 anticoagulation tube, which was dried in an 80° C. oven and stored in a 4° C. refrigerator.

(276) Rats, males, weighing 187 to 197 g, were randomly divided into 2 groups. They started fasting overnight in the afternoon before the experiment but were free to drink water. They were given food 4 hours after administration. The reference compounds (3 mg/kg) were administered to the rats in Group A, and the Example compound (3 mg/kg) was administered to the rats in group B. About 100-200 μL blood was taken from the orbital vein of rats at 15 min, 30 min, 1, 2, 3, 5, 8, and 10 h after administration, and placed in a 0.5 mL Eppendorf tube anti-coagulated with EDTA-K2, which were mixed immediately. After the anti-coagulation, the test tubes were inverted and mixed 5-6 times gently as soon as possible. Then the collected blood was placed in an ice box, and within 30 min, the blood sample was centrifuged at 4000 rpm, 4° C. for 10 min. All of the plasma were collected and stored at −20° C. immediately. After samples were collected at all above time points, the drug concentration in the plasma was determined at each time point.

(277) From the resulting data of mean drug concentration in the plasma verse time after administration, Winnonin software was used to calculate the pharmacokinetics-related parameters of the male SD rats after they were i.g. administered the Example compounds (3 mg/kg) according to the non-compartment statistical moment theory.

(278) Experiments show that the compounds disclosed herein have good activity and pharmacokinetic properties, and are more suitable compounds for inhibiting nucleoside reverse transcriptase, and thus they are suitable for preparing a drug for treating antiviral infection.

(279) It should be understood, these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental methods without specific conditions in the examples generally follow the conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, parts and percentages are parts by weight and weight percent.

(280) The foregoing is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it cannot be assumed that the specific embodiment of the present invention is limited to these descriptions. For ordinary skilled in the art, without departing from the concept of the present invention, several simple deductions or replacements can also be made, which should be deemed to be within the scope of the present invention.