CN114644651A

CN114644651A - Process for producing aryloxyphosphorylated amino acid ester compound

Info

Publication number: CN114644651A
Application number: CN202210389613.3A
Authority: CN
Inventors: 王保刚; 贝祝春; 宋亚彬; 张东娜; 徐力昆; 曹玢旺
Original assignee: Academy of Military Medical Sciences AMMS of PLA
Current assignee: Academy of Military Medical Sciences AMMS of PLA
Priority date: 2022-04-13
Filing date: 2022-04-13
Publication date: 2022-06-21

Abstract

The invention discloses a preparation method of an aryloxy phosphorylated amino acid ester compound, which can be used for synthesizing a ProTide medicament. The synthesis method specifically comprises the steps of reacting (substituted) phenyl dichlorophosphate 1H-1,2, 4-triazole to generate an intermediate product, directly reacting the intermediate product with amino acid ester at normal temperature, then reacting the intermediate product with (substituted) phenol to obtain an aryloxy phosphorylated amino acid ester compound, and continuously recrystallizing to prepare the target aryloxy phosphorylated amino acid ester compound with a single configuration, wherein the aryloxy phosphorylated amino acid ester compound can be further used for synthesizing a ProTide medicament.

Description

Process for producing aryloxyphosphorylated amino acid ester compound

Technical Field

The invention relates to the technical field of medicines, in particular to a method for synthesizing aryloxy phosphorylated amino acid ester, which can be used for preparing a Protide medicine.

Background

Antiviral drugs are powerful weapons for human to overcome viruses, and in the field of antiviral drugs, nucleoside analogs have become one of the major focus areas of drug development since the first synthetic nucleoside drug, uridine, was approved for sale in 1963. Through decades of development, nucleoside drugs have become a class of drugs with important effects in the fields of antitumor and antiviral therapy. At present, dozens of nucleoside drugs are clinically used for treating tumors and viral infections. The mechanism of action of nucleoside drugs is to exert the corresponding biological activity by terminating strand synthesis by inhibition of DNA or RNA polymerase, or by incorporation into nucleic acid strands, after conversion to the triphosphate form in vivo. Due to structural differences from natural nucleosides, nucleoside kinase activity is limited, resulting in low conversion efficiency of the process of monophosphorylation to 5' -monophosphoryl nucleoside, which is often responsible for limiting the activity of nucleoside analogs. The synthesis of nucleoside monophosphates analogs by chemical synthesis is an effective strategy to address this problem.

However, the phosphate group has negative charge under physiological conditions, and is difficult to cross biological membranes, so that the bioavailability of the phosphate group is greatly reduced. The Phosphoramidate (ProTide) prodrug technology developed by Chris McGuigan et al is an effective solution to the above problems. ProTide is derived from PROdrug + nucleoTIDE, and the technical core is that the nucleoside analogue is more conveniently and effectively transmitted into cells in the form of phosphoramidate by a PROdrug mode mainly by utilizing a structure consisting of an aromatic substituent motif and amino acid ester. After entering cells, phosphoramidate is firstly metabolized into a Prodrug (produgs) by esterase, an amino acid group is further removed to form another drug (NMP) of a monophosphate molecule, the NMP is metabolized into Nucleotide Diphosphate (NDP) by NMP-kinase, and then is metabolized into Nucleotide Triphosphate (NTP) by NDP-kinase to finally play a role. By means of the technology, various antiviral nucleoside drugs are approved to be on the market, and Sofosbuvir (Sofosbuvir), a drug for resisting HCV infection, Tenofovir Alafenamide (TAF) and a drug for resisting SARS-CoV-2, namely Remdesivir (GS-5734), are ProTide prodrug technologies. In addition, there are several developed nucleoside drugs based on ProTide technology in clinical research stage.

In order to form chemical caps on nucleosides, which can be enzymatically hydrolyzed, through the development of multi-generation synthesis technology, most of the synthesis of ProTide nucleosides at present needs to synthesize an intermediate compound, which is aryloxy phosphorylated amino acid ester with a leaving group and is used for forming a side chain, such as the following intermediate compounds:

at present, the synthesis method of the compound is to use (substituted) phenoxyl phosphoryl dichloride as a raw material to react with amino acid ester at ultralow temperature (usually-78 ℃) and substituted phenol with an electron-withdrawing substituent to obtain an aryloxy phosphoryl amino acid ester compound containing a leaving group, and then further separate the compound to obtain a target compound with a single configuration. For example, when synthesizing Sofosbuvir, patent WO2010/135569 uses 4-nitrophenyl dichlorophosphate as a raw material, and reacts with phenol at-60 ℃, and then reacts with L-alanine isopropyl ester hydrochloride at-5 ℃ to 0 ℃ to obtain a target product, wherein the specific reaction formula is as follows:

in WO2018/015821, phenyl dichlorophosphate is used as a raw material, and is reacted with L-alanine isopropyl ester hydrochloride at the temperature of-55 ℃, and then is reacted with 5-F phenol at the temperature of 0 ℃ to obtain a target product. The specific reaction formula is as follows

In the synthesis of Reidesvir, WO2016069826 and the thesis Therapeutic efficacy of the small molecule GS-5734against Ebola virus in rhesus monkys (Nature,2016,17,381-385) all react at-78 ℃ with phenyl dichlorophosphate L-alanine 2-ethylbutyl ester hydrochloride, and then react with substituted phenol at 0 ℃ to obtain the target product. The specific reaction formula is as follows.

In addition, in non-patent documents Synthesis of diameteromeric Pure nucleus nucleoside intermediates (J. org. chem.2011,76,8311-.

The existing method for synthesizing the aryloxy phosphorlyated amino acid ester still needs to be improved, and the method has the problems that the reaction activity of the raw material (substituted) phenoxyl phosphoryl dichloride is higher, the reaction needs to be carried out at low temperature for controlling the reaction of the raw material and equivalent reactants, the operation is complex, the requirement on production equipment is higher, and the production cost is high. Therefore, the method for preparing the aryloxy phosphorylated amino acid ester compound which is more economical, efficient, convenient to operate and suitable for large-scale production is important in application value.

Disclosure of Invention

One of the purposes of the invention is to provide a novel preparation method of aryloxy phosphorylated amino acid ester compound, which can improve the synthesis efficiency of the compound, has low experimental condition requirement and convenient operation, avoids low-temperature reaction condition, and can be carried out at room temperature. The invention also aims to provide a preparation method of the aryloxy phosphorylated amino acid ester compound, which has few byproducts, does not need column chromatography purification in all steps, can obtain a single-configuration target product by recrystallization and is suitable for large-scale production.

Specifically, the present invention provides a method for synthesizing an aryloxyphosphorylated amino acid ester represented by the formula (VI), which comprises the following steps in order:

s1, contacting the compound shown in the formula (II) with the compound shown in the formula (III) in the presence of an organic base to obtain the compound shown in the formula (IV);

s2, contacting the compound shown in the formula (IV) with the compound shown in the formula (V) in the presence of a base to obtain the compound shown in the formula (VI);

in the above formulae (II) to (VI),

m and n are each an integer of 0 to 5, m is preferably 0 or 1, n is preferably an integer of 1 to 5,

R¹each independently selected from halogen, nitro, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 haloalkoxy, saturated or partially unsaturated C3-6 cycloalkyl, saturated or partially unsaturated C6-10 heterocyclyl, C6-10 aryl, C6-12 aralkyl, preferably selected from halogen, C1-6 alkyl, C1-6 alkoxy, nitro, C1-6 haloalkyl;

R³each independently selected from halogen, nitro, C1-6 alkyl, C1-6 alkyl halide, C1-6 alkoxy halide, saturated or partially unsaturated C3-6 cycloalkyl, saturated or partially unsaturated C6-10 heterocyclyl, C6-10 aryl, C6-12 aralkyl, preferably selected from halogen, C1-6 alkyl, C1-6 alkoxy, nitro, C1-6 alkyl halide, further preferably selected from nitro and halogen;

R²selected from C1-C10 alkyl, C1-C10 alkenyl, C1-C10 alkynyl, C6-10 aryl, C6-12 aralkyl, preferably selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, tert-butyl, cyclobutyl, cyclopropylmethyl, n-pentyl, isopentyl, neopentyl, cyclopentyl, 3-pentyl, cyclobutylmethyl, n-hexyl, isohexyl, 2-ethylbutyl, cyclohexyl, cyclopentylmethyl, cyclohexylmethyl, phenyl, benzyl;

a is-C (R)⁴)₂A divalent group represented by R⁴Each independently selected from hydrogen, C1-6 alkyl, C6-12 aralkyl; a is preferably-CH (CH)₃)-；

X represents a halogen, preferably Cl or Br.

In the above-mentioned expression,

the expression "-" indicating the position of linkage "indicates the expression of the loop structure in which the linkage site is located at an arbitrary position on the loop structure where the linkage can be formed.

As a preferred embodiment of the present invention, the present invention is to provide a method for synthesizing the following compounds,

in a preferred embodiment of the present invention, the method further comprises the following steps S0 and/or S3:

s0, contacting the compound shown in the formula (I) with 1H-1,2, 4-triazole in the presence of alkali to obtain a compound shown in a formula (II);

in the formula, m and R¹X has the same meaning as that expressed in claim 1,

s3: and (3) dissolving the solid of the compound of the formula (VI) obtained in the step (S2) by using a normal solvent, adding an anti-solvent, uniformly mixing, and recrystallizing to obtain the solid of the compound of the formula (VI).

In a preferred embodiment of the present invention, the step S1 includes the steps of:

dissolving 1-1.5 equivalents of the raw material of the formula (III) in a reaction solvent, stirring at-20-60 ℃, adding 1-1.5 equivalents of organic base into a solution containing 1 equivalent of the compound of the formula (II), slowly dripping the solution containing the formula (III) into the reaction solution, and continuously keeping the temperature at-20-60 ℃ for reacting for 0.5-12 hours to obtain a solution containing the compound of the formula (IV).

The solvent is dry diethyl ether or THF or CH₂Cl₂Or CH₃The organic base in the CN, S1 step is selected from triethylamine, diisopropylethylamine, triisopropylamine and triallyl amineOrganic bases of amines, diallylethylamines, pyridine, 1-methylimidazole, 4-dimethylaminopyridine, 2, 6-lutidine, 2, 4-lutidine, preferably 1-methylimidazole.

In a preferred embodiment of the present invention, the step S2 includes the steps of:

adding 1-2 equivalents of alkali into the solution containing the compound shown in the formula (IV) obtained in the step S1 at the temperature of-20-60 ℃, slowly dripping the solution obtained by dissolving 0.8-1.5 equivalents of the compound shown in the formula (V) in the solvent while stirring into the solution, reacting for 0.5-12 hours, removing the solvent, extracting by using ethyl acetate and water to obtain a crude product of the compound shown in the formula (VI),

the solvent is dry diethyl ether or THF or CH₂Cl₂Or CH₃CN, wherein the base is selected from triethylamine, diisopropylethylamine, triisopropylamine, triallylamine, diallyl ethylamine or pyridine.

In a preferred embodiment of the present invention, the step S0 includes the steps of:

adding 2-6 equivalents of alkali into a solution of 1H-1,2, 4-triazole at-20-60 ℃, uniformly stirring, dropwise adding 1-1.5 equivalents of a compound shown in formula (I) into a reaction solution at the temperature, reacting for 0.5-6H, and filtering to obtain a solution containing a compound shown in formula (II), wherein the solvent is dry diethyl ether, THF or CH₂Cl₂Or CH₃CN in the S0, the base is triethylamine, diisopropylethylamine, triisopropylamine, triallylamine, diallylethylamine or pyridine.

In a preferred embodiment of the present invention, in the step S1, the compound represented by the formula (II) and the compound represented by the formula (III) are mixed at 0 to 4 ℃, and the reaction is continued while slowly raising the temperature to room temperature, and further preferred reaction conditions are: adding each reagent at 0 ℃, heating to room temperature for reaction for 1-2 hours; in the step S0, the reaction is carried out at 20 to 25 ℃.

In a preferred embodiment of the present invention, the reaction is continuously carried out by sequentially carrying out the steps S0, S1 and S2, and the reaction solution after each step is used as it is for the reaction in the next step,

the charging ratio of each reactant is calculated by the mass: 1H-1,2, 4-triazole, a compound represented by the formula (III), an organic base, a compound represented by the formula (V), 1-1.5: 2-3: 1: 1-1.5: 0.8-1.5.

In a preferred embodiment of the present invention, in the step S3, the normal solvent is selected from the group consisting of ethyl acetate, isopropyl acetate, butyl acetate, isopropyl ether, diethyl ether, methyl tert-butyl ether, THF, CH₂Cl₂2-methyltetrahydrofuran or 1, 4-dioxane, in a ratio of the n-solvent to the solid of the compound of formula (VI) obtained in the step S2: adding 0.2-5g of a solid of the compound shown in the formula (VI) into 1mL of a normal solvent, wherein the anti-solvent is one or a mixed solvent of any one of n-pentane, n-hexane, n-heptane, cyclopentane, cyclohexane, cycloheptane and petroleum ether in any proportion, standing at the temperature of-10-40 ℃ to separate out a solid, and the proportion of the anti-solvent to the normal solvent is 2-10: 1.

in a preferred embodiment of the invention, ethyl acetate is used as a normal solvent in the recrystallization process, the anti-solvent is n-hexane or petroleum ether, the ratio of the anti-solvent to the normal solvent is 3.5-4.5: 1, and the mixture is kept stand at the temperature of 4-25 ℃ to precipitate a solid.

Compared with the prior art, the method for synthesizing and preparing the aryloxy phosphorylated amino acid ester has at least the following beneficial effects: the 1H-1,2, 4-triazole is used for reducing the reactivity of the (substituted) phenoxyphosphoryl dichloride of raw materials, so that the reaction of the (substituted) phenoxyphosphoryl dichloride with amino acid ester and aryloxy leaving groups can be carried out at room temperature in sequence through simple stoichiometric control, and the whole process can be continuously carried out without post-treatment. According to the invention, column chromatography is not required in the reaction process, the required single-configuration product can be obtained by recrystallization, the compound shown in the formula (VI) can be quickly and effectively prepared, the total yield of the reaction is improved, the cost of the final product is effectively reduced, and meanwhile, the method has good experimental operability and can realize large-scale production. Meanwhile, the final product can be easily recrystallized to obtain a high-purity solid, and the method has important significance for the synthesis of the Protide medicines.

Detailed Description

The respective elements of the present invention will be described in more detail below.

Definition of

Unless defined otherwise below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. Reference to the techniques used herein is intended to refer to those techniques commonly understood in the art, including those variations of or alternatives to those techniques that would be apparent to those skilled in the art. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

The term "contacting" as used in the present invention is to be understood in a broad sense and may be any means which enables a chemical reaction of at least two reactants, e.g. mixing of two reactants under suitable conditions. The reactants to be contacted may be mixed with stirring as necessary, and thus, the type of stirring is not particularly limited, and may be, for example, mechanical stirring, that is, stirring under the action of a mechanical force.

In the present invention, "a compound represented by the formula N" is also sometimes referred to herein as "compound N", and N is any integer of 1 to 4 herein, for example, "a compound represented by the formula 2" may also be referred to herein as "compound 2".

In the present invention, the terms "first", "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or as implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.

The terms "comprising," "including," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

As used herein, the term "alkylene" denotes a saturated divalent hydrocarbon group, preferably a saturated divalent hydrocarbon group having 1,2, 3,4,5 or 6 carbon atoms, such as methylene, ethylene, propylene or butylene.

The term "alkyl" as used herein is defined as a linear or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl group has 1 to 12, e.g., 1 to 6, carbon atoms. For example, as used herein, the term "C1-6 alkyl" refers to a linear or branched group of 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, or n-hexyl) optionally substituted with 1 or more (such as 1 to 3) suitable substituents such as halogen (when the group is referred to as "haloalkyl") (e.g., CH)₂F、CHF₂、CF₃、CCl₃、C₂F₅、C₂Cl₅、CH₂CF₃、CH₂Cl or-CH₂CH₂CF₃Etc.). The term "C1-4 alkyl" refers to a linear or branched aliphatic hydrocarbon chain of 1 to 4 carbon atoms (i.e., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl).

As used herein, the term "alkenyl" means a linear or branched monovalent hydrocarbon radical containing one double bond and having from 2 to 6 carbon atoms ("C)_2-6Alkenyl "). The alkenyl group is, for example, vinyl, 1-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl and 4-methyl-3-pentenyl. When the compounds of the invention contain an alkenyl group, the compounds may be present in pure E (entgegen) form, in pure Z (zusammen) form or in any mixture thereof.

As used herein, the term "alkynyl" denotes a monovalent hydrocarbon group containing one or more triple bonds, preferably having 2,3,4,5 or 6 carbon atoms, such as ethynyl or propynyl.

As used herein in the context of the present invention,the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic, including spiro, fused or bridged systems (such as bicyclo [ 1.1.1)]Pentyl, bicyclo [2.2.1]Heptyl, bicyclo [3.2.1]Octyl or bicyclo [5.2.0]Nonyl, decalinyl, etc.)), optionally substituted with 1 or more (such as 1 to 3) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term "C_3-6Cycloalkyl "refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring of 3 to 6 ring-forming carbon atoms (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) optionally substituted with 1 or more (such as 1 to 3) suitable substituents, for example, methyl-substituted cyclopropyl.

As used herein, the terms "cycloalkylene", "cycloalkyl" and "hydrocarbon ring" refer to saturated (i.e., "cycloalkylene" and "cycloalkyl") or unsaturated (i.e., having one or more double and/or triple bonds within the ring) monocyclic or polycyclic hydrocarbon rings having, for example, 3 to 10 (suitably 3 to 8, more suitably 3 to 6) ring carbon atoms, including, but not limited to, (trimethylene) cyclopropyl (ring), (tetramethylene) cyclobutyl (ring), (tetramethylene) cyclopentyl (ring), (hexamethylene) cyclohexyl (ring), (tetramethylene) cycloheptyl (ring), (hexamethylene) cyclooctyl (ring), (hexamethylene) nonyl (ring), (hexamethylene) cyclohexenyl (ring), and the like.

As used herein, the terms "heterocyclyl", "heterocyclylene" and "heterocycle" refer to saturated (i.e., heterocycloalkyl) or partially unsaturated (i.e., having one or more double and/or triple bonds within the ring) cyclic groups having, for example, 3-10 (suitably 3-8, more suitably 3-6) ring atoms, wherein at least one ring atom is a heteroatom selected from N, O and S and the remaining ring atoms are C. For example, a "3-to 10-membered (sub) heterocyclic (yl)" is a saturated or partially unsaturated (sub) heterocyclic (yl) group having 2-9 (e.g., 2,3,4,5,6, 7, 8 or 9) ring carbon atoms and one or more (e.g., 1,2, 3 or 4) heteroatoms independently selected from N, O and S. Examples of heterocyclylene and heterocycle include, but are not limited to: oxirane (ene), aziridine (ene), azetidine (azetidinyl) (ene), oxetane (oxirane), (tetramethylene) tetrahydrofurane, (tetramethylene) dioxolyl (dioxalinyl), (tetramethylene) pyrrolidinyl, (tetramethylene) pyrrolidinone, (tetramethylene) imidazolidine, (tetramethylene) pyrazolene, (tetramethylene) pyrrolinyl, (tetramethylene) tetrahydropyranyl, (tetramethylene) piperidene, (tetramethylene) morpholinyl, (tetramethylene) dithianyl, (tetramethylene) thiomorpholinyl, (tetramethylene) piperazinyl, or trithianyl (trithianyl). The group also encompasses bicyclic ring systems, including spiro, fused or bridged systems (such as 8-azaspiro [4.5] decane, 3, 9-diazaspiro [5.5] undecane, 2-azabicyclo [2.2.2] octane, and the like). The heterocyclylene and heterocyclic ring(s) may be optionally substituted with one or more (e.g. 1,2, 3 or 4) suitable substituents.

As used herein, the terms "(arylene) group" and "aromatic ring" refer to an all-carbon monocyclic or fused ring polycyclic aromatic group having a conjugated pi-electron system. For example, as used herein, the term "C_6-10(arylene) and C_6-10Aromatic ring "means an aromatic group containing 6 to 10 carbon atoms, such as (phenylene) phenyl (benzene ring) or (phenylene) naphthyl (naphthalene ring). The aryl (ene) and aromatic rings are optionally substituted with 1 or more (such as 1 to 3) suitable substituents (e.g. halogen, -OH, -CN, -NO)₂、C_1-6Alkyl, etc.).

As used herein, the terms "(arylene) heteroaryl" and "heteroaryl ring" refer to a monocyclic, bicyclic or tricyclic aromatic ring system having 5,6, 8, 9, 10, 11,12, 13 or 14 ring atoms, in particular 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and which contains at least one heteroatom which may be the same or different (said heteroatom being, for example, oxygen, nitrogen or sulfur), and which, in addition, may be benzo-fused in each case. In particular, "(arylene) heteroaryl" or "heteroaromatic ring" is selected from thienyl (ene), furyl (ene), pyrrolyl (ene), oxazolyl (ene), thiazolyl (ene), imidazolyl (ene), pyrazolyl (ene), isoxazolyl (ene), isothiazolyl (ene), oxadiazolyl (ene), triazolyl (ene), thiadiazolyl (ene), and the like, and benzo derivatives thereof; or pyridyl (ene), pyridazinyl (ene), pyrimidinyl (ene), pyrazinyl (ene), triazinyl (ene), etc., and benzo derivatives thereof.

As used herein, the term "aralkyl" preferably denotes aryl or heteroaryl substituted alkyl, wherein the aryl, heteroaryl and alkyl are as defined herein. Typically, the aryl group can have 6 to 14 carbon atoms, the heteroaryl group can have 5 to 14 ring atoms, and the alkyl group can have 1 to 6 carbon atoms. Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, phenylbutyl.

As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

As used herein, the term "nitrogen-containing heterocycle" refers to a saturated or unsaturated monocyclic or bicyclic group having 2,3,4,5,6, 7, 8, 9, 10, 11,12, or 13 carbon atoms and at least one nitrogen atom in the ring, which may also optionally contain one or more (e.g., one, two, three, or four) selected from N, O, C ═ O, S, S ═ O and S (═ O)₂Is attached to the remainder of the molecule through the nitrogen atom of the nitrogen-containing heterocycle and any remaining ring atom, the nitrogen-containing heterocycle is optionally benzo-fused, and is preferably attached to the remainder of the molecule through the nitrogen atom of the nitrogen-containing heterocycle and any carbon atom of the fused benzene ring.

If a substituent is described as being "independently selected from" a group, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

Unless indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

When a bond of a substituent is shown through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

The invention also includes all pharmaceutically acceptable isotopically-labeled compounds, which are identical to those of the present invention, except that one or more atoms are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number prevailing in nature. Fitting bagExamples of isotopes that can be incorporated into compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium (g), (b), (c), (d) and (d)²H) Tritium (a)³H) ); isotopes of carbon (e.g. of¹¹C、¹³C and¹⁴C) (ii) a Isotopes of chlorine (e.g. of³⁶Cl); isotopes of fluorine (e.g. of fluorine)¹⁸F) (ii) a Isotopes of iodine (e.g. of iodine)¹²³I and¹²⁵I) (ii) a Isotopes of nitrogen (e.g. of¹³N and¹⁵n); isotopes of oxygen (e.g. of¹⁵O、¹⁷O and¹⁸o); isotopes of phosphorus (e.g. of phosphorus)³²P); and isotopes of sulfur (e.g. of³⁵S). Certain isotopically-labeled compounds of the present invention (e.g., those into which a radioisotope is incorporated) are useful in drug and/or substrate tissue distribution studies (e.g., assays). Radioisotope tritium (i.e. tritium)³H) And carbon-14 (i.e., 14C) are particularly useful for this purpose because of their ease of incorporation and ease of detection. With positron-emitting isotopes (e.g.)¹¹C、¹⁸F、¹⁵O and¹³n) can be used to examine substrate receptor occupancy in Positron Emission Tomography (PET) studies. Isotopically labeled compounds of the present invention can be prepared by processes analogous to those described in the accompanying schemes and/or in the examples and preparations by using an appropriate isotopically labeled reagent in place of the non-labeled reagent employed previously. Pharmaceutically acceptable solvates of the invention include those in which the crystallization solvent may be isotopically substituted, e.g., D₂O, acetone-d₆Or DMSO-d₆。

The term "stereoisomer" denotes an isomer formed as a result of at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, they can give rise to racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Certain individual molecules may also exist as geometric isomers (cis/trans). Similarly, the compounds of the invention may exist as mixtures of two or more structurally different forms (commonly referred to as tautomers) in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It is understood that the scope of this application encompasses all such isomers or mixtures thereof in any ratio (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

Solid lines may be used herein

Solid wedge shape

Or virtual wedge shape

Chemical bonds of the compounds of the present invention are depicted. The use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers (e.g., particular enantiomers, racemic mixtures, etc.) at that carbon atom are included. The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the stereoisomers shown are present. When present in a racemic mixture, solid and dotted wedges are used to define the relative stereochemistry, not the absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention are intended to exist as stereoisomers, including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotamers, conformers, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerization and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

As used herein, the term "ester" means an ester derived from a compound of the respective formula in the present application, including physiologically hydrolysable esters (which can be hydrolysed under physiological conditions to release the compound of the invention in the form of a free acid or alcohol). The compounds of the invention may themselves also be esters.

The invention also encompasses compounds of the invention containing a protecting group. In any process for preparing the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved by conventional protecting Groups, such as those described in T.W.Greene & P.G.M.Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. The protecting group may be removed at a suitable subsequent stage using methods known in the art.

The term "about" means within. + -. 10%, preferably within. + -. 5%, more preferably within. + -. 2% of the stated value.

Synthesis method of the invention

The invention provides a method for synthesizing aryloxy phosphorylated amino acid ester shown in formula (VI), which is characterized by sequentially comprising the following steps:

in the above formulae (II) to (VI),

m and n are integers of 0 to 5, m is preferably 0 or 1, n is preferably an integer of 1 to 5,

R¹each independently selected from the group consisting of halogen, nitro, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 haloalkoxy, saturated or partially unsaturated C3-6 cycloalkyl, saturated or partially unsaturated C6-10 heterocyclyl, and mixtures thereof,C6-10 aryl, C6-12 aralkyl, preferably selected from halogen, C1-6 alkyl, C1-6 alkoxy, nitro, C1-6 haloalkyl;

X represents a halogen, preferably Cl or Br.

In the above-mentioned expression,

in the present invention, it is the starting compound (the compound of formula (II)) in the S1 step that has appropriate reactivity, so that the reaction with the amino acid ester and the aryloxy leaving group can be performed sequentially at room temperature by simple stoichiometric control, and the reaction rate is much faster without requiring a complicated low-temperature operation environment.

In addition, the compound of formula (II) can be simply prepared by using the compound of formula (I) and 1H-1,2, 4-triazole through the following S0 process, when the compound of formula (I) is (substituted) phenoxyphosphoryl dichloride, the synthetic method of the invention is equivalent to that the compound is completely compatible with the raw materials for synthesizing the intermediate compound shown in formula (VI) in the existing pharmaceutical industry, thereby utilizing the existing supplier resources and fundamentally solving the problem of easy obtaining of the raw materials.

s0, contacting the compound shown as the formula (I) with 1H-1,2, 4-triazole in the presence of alkali to obtain a compound shown as a formula (II);

in the formula, m and R¹X has the same meaning as that expressed in claim 1,

The base may be an organic base or an inorganic base, the inorganic base may be selected from potassium hydroxide, sodium hydrogen carbonate, sodium carbonate, potassium hydrogen carbonate, potassium carbonate, cesium carbonate, potassium phosphate, potassium dihydrogen phosphate, and dipotassium hydrogen phosphate, and the organic base may be selected from triethylamine, diisopropylethylamine, triisopropylamine, diallylethylamine, and pyridine. In the present invention, an organic base is preferably used, and triethylamine is more preferably used.

dissolving 1-1.5 equivalents of the raw material of the formula (III) in a reaction solvent, stirring at-20-60 ℃, adding 1-1.5 equivalents of organic base into a solution containing 1 equivalent of the compound of the formula (II), slowly dripping the solution containing the formula (III) into the reaction solution, continuously keeping the temperature at-20-60 ℃ for reaction for 0.5-12 hours to obtain a solution containing the compound of the formula (IV),

the solvent is dry diethyl ether or THF or CH₂Cl₂Or CH₃The organic base in the CN, S1 step is an organic base selected from triethylamine, diisopropylethylamine, triisopropylamine, triallylamine, diallylethylamine, pyridine, 1-methylimidazole, 4-dimethylaminopyridine, 2, 6-dimethylpyridine, 2, 4-dimethylpyridine, preferably 1-methylimidazole.

in the present invention, as the solvent, organic solvents known in the art for chemical reactions can be used, and examples thereof include DMSO, DMF, tetrahydrofuran, halogenated hydrocarbon solvents, alkyl alcohol solvents of C1 to C6, ester solvents, benzene, toluene, pyridine, nitrile solvents, THF, and the like. The halogenated hydrocarbon solvent is a saturated or unsaturated chlorinated hydrocarbon having 1 or 2 carbon atoms, and is usually selected from the group consisting of dichloromethane, trichloromethane, carbon tetrachloride, 1, 1-dichloroethane, 1, 2-dichloroethane, 1,1, 1-trichloroethane, 1,1, 2-trichloroethane, 1,1, 1, 2-tetrachloroethane, 1,1, 2, 2-tetrachloroethane, pentachloroethane, 1, 1-dichloroethylene, 1, 2-dichloroethylene, trichloroethylene and tetrachloroethylene. More preferably dichloromethane, trichloromethane, 1,1, 1-trichloroethane, trichloroethylene and tetrachloroethylene, and further more preferablyPreferred are dichloromethane and trichloromethane. The alkanol solvent of C1 to C6 may be methanol, ethanol, isopropanol, n-butanol, cyclohexanol, etc., which are commonly used, but is not limited thereto. Examples of the ester solvent include, but are not limited to, ethyl acetate, methyl acetate, and ethyl formate. Preferably, the solvent used is dry diethyl ether or THF or CH₂Cl₂Or CH₃CN, further preferably dried diethyl ether or THF, most preferably THF.

adding 2-6 equivalents of alkali into a solution of 1H-1,2, 4-triazole at-20-60 ℃, uniformly stirring, dropwise adding 1-1.5 equivalents of a compound shown in formula (I) into a reaction solution at the temperature, reacting for 0.5-6H, and filtering to obtain a solution containing a compound shown in formula (II), wherein the solvent is dry diethyl ether, THF or CH₂Cl₂Or CH₃CN。

In a preferred embodiment of the present invention, the compound represented by the formula (II) and the compound represented by the formula (III) are mixed at 0 to 4 ℃ in the step S1, the reaction is continued while slowly raising the temperature to room temperature, and the reaction is continued at 20 to 25 ℃ in the step S0.

the charging ratio of each reactant is calculated by the mass: 1H-1,2, 4-triazole, a compound represented by formula (I), an organic base (1-methylimidazole), a compound represented by formula (V), 1-1.5: 2-3: 1:1 to 1.5:0.8 to 1.5, and the most preferable ratio of the inventions is 1.05: 2.2: 1:1:1.

That is, the most preferred combination of the synthetic methods of the present invention can be summarized as the following synthetic route:

in the inventionIn a preferred embodiment, in the step S3, the n-solvent is selected from ethyl acetate, isopropyl acetate, butyl acetate, isopropyl ether, diethyl ether, methyl tert-butyl ether, THF, CH₂Cl₂2-methyltetrahydrofuran or 1, 4-dioxane, in a ratio of the n-solvent to the solid of the compound of formula (VI) obtained in the step S2: adding 0.2-5g of a solid of the compound shown in the formula (VI) into 1mL of a normal solvent, wherein the anti-solvent is one or a mixed solvent of any one of n-pentane, n-hexane, n-heptane, cyclopentane, cyclohexane, cycloheptane and petroleum ether in any proportion, standing at the temperature of-10-40 ℃ to separate out a solid, and the proportion of the anti-solvent to the normal solvent is 2-10: 1.

Specific examples of preferred embodiments of the present invention include the following continuous processes:

adding 2-3 equivalents of 1H-1,2, 4-triazole into a reaction bottle at-20-60 ℃, and adding dried diethyl ether or THF or CH₂Cl₂Or CH₃CN, stirring, adding 2-6 equivalents of triethylamine or diisopropylethylamine or triisopropylamine or triallylamine or diallyl ethylamine or pyridine, dropwise adding 1-1.5 equivalents of the compound of formula (I) into a reaction bottle at the temperature, reacting for 0.5-6h, filtering after the reaction is finished, washing a filter cake twice by using a reaction solvent, combining the filtrates, stirring at-20-60 ℃, adding 1-1.5 equivalents of organic base, dissolving 1 equivalent of the compound of formula (III) into the reaction solvent, slowly dropwise adding the solution into the reaction solution at the temperature, continuing the reaction for 0.5-12h after the dropwise adding is finished, adding 1-2 equivalents of triethylamine or diisopropylethylamine or triisopropylamine or triallylamine or diallyl ethylamine or pyridine at-20-60 ℃, dissolving 0.8-1.5 equivalents of the compound of formula (V) into the reaction solvent, dropwise adding the solution into the reaction solution, the reaction is continued for 0.5 to 12 hours. After the reaction is finished, evaporating partial reaction solvent, adding ethyl acetate to dissolve, washing, drying, filtering to remove drying agent, evaporating solvent, adding residual solventThe mass (g) of the residue is 1-2 times of the volume (mL) of n-solvent ethyl acetate or isopropyl acetate or butyl acetate or isopropyl ether or ethyl ether or methyl tert-butyl ether or THF or CH₂Cl₂Or dissolving the residue in 2-methyltetrahydrofuran or 1, 4-dioxane, adding one or a mixture of more of n-pentane, n-hexane, n-heptane, cyclopentane, cyclohexane, cycloheptane or petroleum ether which are anti-solvents in an amount of 2-10 times the volume of the n-solvent, uniformly mixing, standing at-10-40 ℃ to separate out a solid, filtering, washing and drying to obtain the product of the general formula (VI).

The synthesis method of the present invention can be modified based on the known techniques without departing from the concept of the present invention, and such modifications are also included in the scope of the present invention.

Examples

These examples are not intended to limit the scope of the present invention, but are merely provided to illustrate the respective elements of the present invention in more detail.

The structure of the compound is determined by nuclear magnetic resonance spectrum (¹H NMR) and/or Mass Spectrometry (MS).

Chemical shifts (δ) are given in parts per million (ppm).¹The H NMR measurements were carried out on a Bruker 400 or Bruker 600 or Varian 300 nuclear magnetic instrument, the test solvent being deuterated methanol (CD)₃OD), deuterated chloroform (CDCl)₃) Or hexadeutero dimethyl sulfoxide (DMSO-d)₆) Internal standard is Tetramethylsilane (TMS).

LC-MS measurements were performed on an Agilent LC-MS-1110 LC-MS, an Agilent LC-MS-6110 LC-MS, an Agilent LC-MS-6120 LC-MS (manufacturer: Agilent) or Shimadzu LC-MS 2020.

Preparative high performance liquid chromatography was performed using an MS triggered automatic purification system (Waters), Gilson GX-281(Gilson) or semi-preparative liquid chromatograph (innovative universal LC3000(Ddlsogel, C18, 30mm x 250mm 10 μm)).

Thin Layer Chromatography (TLC) was carried out using a yellow sea brand HSGF 254 (5X 20cm) silica gel plate, and preparative thin layer chromatography was carried out using a silica gel plate of GF 254(0.4 to 0.5nm) produced by Nicotine.

The reaction is detected by Thin Layer Chromatography (TLC) or LC-MS, and the developing agent system including dichloromethane and methanol system, n-hexane and ethyl acetate system, and petroleum ether and ethyl acetate system is adjusted according to the polarity of the compound to be separated (by adjusting the volume ratio of the solvent or adding triethylamine, etc.).

As is not specifically shown in the examples, the reaction temperature is room temperature (20 ℃ C. to 30 ℃ C.).

The reagents used in the examples were purchased from Aldrich Chemical Company, Shanghai national reagent science and technology, Inc., Bailingwei reagent, Shaoshi technology (Shanghai), Inc., or Aiberkojic, Inc.

Adding 1.520g (22.0mmol) of 1H-1,2, 4-triazole into a reaction bottle with a magnetic stirring bar at room temperature, adding 30.0mL of dry THF and 2.226g (22.0mmol) of triethylamine, dissolving 2.215g (10.5 mmol) of phenyl dichlorophosphate in 5.0mL of THF, dropwise adding the mixture into the reaction bottle under magnetic stirring, reacting at room temperature for 1.0H, filtering under reduced pressure to remove precipitates, washing a filter cake with 3.0mL of THF (multiplied by 2), combining the filtrates, adding 0.821g (10.0mmol) of organic base, magnetically stirring at 0 ℃, dissolving 1.733g (10.0mmol) of L-alanine (2-ethylbutyl) ester in 10.0 mL of THF, slowly dropwise adding the mixture into the reaction solution, gradually raising the temperature to room temperature after dropwise adding, reacting at room temperature for 2.0H, continuously adding 1.113g (11.0mmol) of triethylamine into the reaction bottle, and adding 1.841g (10.0mmol) of 2,3,4,5, 6-pentafluorophenol in 0.5mL of THF, the reaction was continued at room temperature for 8.0 h. After the reaction, most of the solvent was evaporated under reduced pressure, and 50.0mL of ethyl acetate was added to dissolve the solvent, and 10% NaH was added to the solution in a separatory funnel₂PO₄The solution was washed 1 time with 50.0mL of saturated NaHCO₃Washing with saturated NaCl solution once, and washing with anhydrous Na as organic layer₂SO₄Drying, filtering to remove desiccant, evaporating to remove solvent, adding ethyl acetate 2.0mL and n-hexane 8.0mL to dissolve the residue, standing at 4 deg.C overnight to precipitate crystal, and filtering to obtain target product [ (2s) - (4-nitro-benzene)Oxy) -phenoxy-phosphoryl]-L-alanine (2-ethylbutyl) ester.

Nuclear magnetism of it¹H NMR(600MHz,CDCl₃) δ 0.91(t, J ═ 7.2Hz,6H),1.34-1.39(m, 4H),1.49(d, J ═ 7.2Hz,3H),1.53-1.57(m,1H),4.03(dd, J ═ 12.0,9.6Hz,1H), 4.07-4.12(m,2H),4.18-4.25(m,1H),7.23-7.39(m, 5H); nuclear magnetism¹³C NMR(125 MHz,CDCl₃) δ 173.16,173.10,150.15,150.10,129.83,125.65,125.64,120.06, 120.03,67.83,50.54,50.53,40.26,23.18,23.16,21.09,21.06,10.94, 10.92; nuclear magnetic resonance³¹P NMR(240MHz,CDCl₃) Delta-1.62; high resolution mass spectrum HRMS (ESI) calcd.for C₂₁H₂₄F₅NO₅P⁺[M+H]⁺:496.1307,found:496.1296。

Example 2: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine isopropyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.312g (10.0mmol) of isopropyl L-alanine to give the desired product, [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -isopropyl L-alanine.

Nuclear magnetism of¹H NMR(600MHz,CDCl₃) δ 1.25(t, J ═ 7.8Hz,6H),1.46(d, J ═ 7.2Hz,3H),3.94 to 4.00(m,1H),4.01(t, J ═ 10.2Hz,2H),4.11 to 4.17(m,1H), 5.02 to 5.06(m,1H),7.20 to 7.37(m, 5H); nuclear magnetism¹³C NMR(125MHz,CDCl₃) δ 172.48, 172.42,150.17,150.12,129.83,125.63,120.06,120.03,69.60,50.61,21.62, 21.59,20.96, 20.93; nuclear magnetism³¹P NMR(240MHz,CDCl₃) Delta-1.59; high resolution mass spectrum HRMS (ESI) calcd.for C₁₈H₁₈F₅NO₅P⁺[M+H]⁺:454.0837,found:454.0819。

Example 3: preparation of [ (2s) - (4-Nitro-phenoxy) -phenoxy-phosphoryl ] -L-alanine (2-ethylbutyl) ester

The procedure is as in example 1 except that 2,3,4,5, 6-pentafluorophenol in the starting material is replaced with 1.391g (10.0mmol) of 4-nitrophenol to obtain the desired product [ (2s) - (4-nitro-phenoxy) -phenoxy-phosphoryl ] -L-alanine (2-ethylbutyl) ester.

Nuclear magnetism of¹H NMR(600MHz,CDCl₃) δ 0.87(t, J ═ 7.2Hz,6H),1.29-1.35(m, 4H),1.41(d, J ═ 7.2Hz,3H),1.47-1.53(m,1H),3.97(dd, J ═ 11.4,9.6Hz, 1H),4.04(qd, J ═ 10.8,6.0Hz,2H),4.13-4.19(m,1H),7.19-7.24(m,3H), 7.35(t, J ═ 7.8Hz,2H),7.40(d, J ═ 9.0Hz, 2H); nuclear magnetism¹³C NMR(125MHz,CDCl₃) δ 173.17,173.11,155.60,155.56,150.23,150.19,144.67,129.88,125.64, 125.53,120.82,120.78,120.13,120.10,67.84,50.54,50.52,45.79,40.23, 23.18,23.15,21.16,21.13,10.96,10.93, 8.63; nuclear magnetic resonance³¹P NMR(240MHz,CDCl₃) Delta-3.16; high resolution mass spectrum HRMS (ESI) calcd.for C₂₁H₂₈N₂O₇P⁺[M+H]⁺:451.1629,found: 451.1617。

Example 4: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine methyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with L-alanine methyl ester 1.031g (10.0mmol) to obtain the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine methyl ester.

Nuclear magnetism of it¹H NMR(400MHz,CDCl₃) δ 1.47(d, J ═ 7.2Hz,3H),3.74(s,3H), 3.90-3.96(m,1H),4.17-4.23(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz, CDCl₃) δ 173.45,173.36,150.21,150.14,129.87,125.68,120.07,120.02, 52.71,50.49,50.47,20.96, 20.92; nuclear magnetism³¹P NMR (160MHz, CDCl3): delta-1.76; high resolution mass spectrum HRMS (ESI) calcd.for C₁₆H₁₃F₅NO₅PNa⁺[M+Na]⁺:448.0344,found: 448.0341。

Example 5: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine ethyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with L-alanine ethyl ester 1.171g (10.0mmol) to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine ethyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 1.27(t, J ═ 7.2Hz,3H),1.47(d, J ═ 7.2Hz,3H),3.92-3.97(m,1H),4.14-4.22(m,3H),7.19-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 172.98,172.89,150.23,150.16,129.87,125.67, 120.08,120.03,61.86,50.54,21.00,20.95, 14.09; nuclear magnetic resonance³¹P NMR(160MHz,CDCl₃) Delta-1.69; high resolution mass spectrum HRMS (ESI) calcd.for C₁₇H₁₅F₅NO₅PNa⁺[M+Na]⁺:462.0500,found: 462.0496。

Example 6: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine n-propyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with 1.312g (10.0mmol) of N-propyl L-alanine to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -N-propyl L-alanine.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.94(t, J ═ 7.2Hz,3H),1.47(d, J ═ 6.4Hz,3H),1.62 to 1.71(m,2H),3.94 to 4.00(m,1H),4.10, (t, J ═ 6.4Hz,2H), 4.14 to 4.24(m,1H),7.20 to 7.38(m, 5H); nuclear magnetic resonance¹³C NMR(100MHz,CDCl₃) δ 173.08, 173.00,150.23,150.16,129.87,125.67,120.09,120.04,67.42,50.57,50.55, 21.89,21.07,21.03, 10.22; nuclear magnetism³¹P NMR(160MHz,CDCl₃) Delta-1.66; high resolution mass spectrum HRMS (ESI) calcd.for C₁₈H₁₈F₅NO₅P⁺[M+H]⁺:454.0837,found:454.0837。

Example 7: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine n-butyl ester

The procedure is as in example 1, except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.452g (10.0mmol) of n-butyl L-alanine to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -N-butyl L-alanine.

Nuclear magnetism of it¹H NMR(400MHz,CDCl₃) δ 0.93(t, J ═ 7.2Hz,3H),1.32-1.42(m, 2H),1.47(d, J ═ 6.8Hz,3H),1.58-1.65(m,2H),3.92-3.98(m,1H),4.14(t, J ═ 6.8Hz,2H),4.15-4.21(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz, CDCl₃) δ 173.08,172.99,150.22,150.15,129.86,125.67,120.09,120.04, 65.73,50.57,50.55,30.53,21.06,21.02,19.00, 13.61; nuclear magnetic resonance³¹P NMR(160MHz, CDCl₃) Delta-1.68; high resolution mass spectrum HRMS (ESI) calcd.for C₁₉H₁₉F₅NO₅PNa⁺[M+Na]⁺: 490.0813,found:490.0877。

Example 8: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine isobutyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with 1.452g (10.0mmol) of isobutyl L-alanine to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -isobutyl L-alanine.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.93(d, J ═ 6.8Hz,6H),1.48(d, J ═ 7.2Hz,3H),1.89-2.00(m,1H),3.89-3.99(m,3H),4.16-4.25(m,1H),7.19-7.38 (m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) Delta 173.06,172.97,150.22,150.15,129.87, 125.68,120.09,120.04,71.85,50.57,50.55,27.71,21.13,21.09,18.92 and 18.90; nuclear magnetic resonance³¹P NMR(160MHz,CDCl₃) Delta-1.66; high resolution mass spectrum HRMS (ESI) calcd.for C₁₉H₁₉F₅NO₅PNa⁺[M+Na]⁺:490.0813,found:490.0892。

Example 9: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine tert-butyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.452g (10.0mmol) of tert-butyl L-alanine to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -tert-butyl L-alanine.

Nuclear magnetism of it¹H NMR(400MHz,CDCl₃) δ 0.94(t, J ═ 7.2Hz,3H),1.47(d, J ═ 6.4Hz,3H),1.62 to 1.71(m,2H),3.94 to 4.00(m,1H),4.10, (t, J ═ 6.4Hz,2H), 4.14 to 4.24(m,1H),7.20 to 7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) Delta 172.16, 172.07,150.27,150.20,129.85,125.62,120.10,120.05,82.48,51.05,51.03, 27.89,21.07, 21.03; nuclear magnetic 31P NMR (160MHz, CDCl)₃) Delta-1.48; high resolution mass spectrum HRMS (ESI) calcd.for C₁₉H₁₉F₅NO₅PNa⁺[M+Na]⁺:490.0813,found:490.0873。

Example 10: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclobutyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with 1.432g (10.0mmol) of L-alanine cyclobutyl ester to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclobutyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 1.46(d, J ═ 7.2Hz,3H),1.60-1.69(m, 1H),1.82(q, J ═ 10.4Hz,1H),2.00-2.12(m,2H),2.34-2.36(m,2H),3.93 (t, J ═ 10.4Hz,1H),4.10-4.20(m,1H),4.97-5.04(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 172.29,172.21,150.23,150.16,139.23,129.87, 125.66,120.08,120.03,70.18,50.47,50.46,30.16,30.14,20.95,20.90, 13.42; nuclear magnetism³¹P NMR(160MHz,CDCl₃) Delta-1.69; high resolution mass spectrum HRMS (ESI) calcd.for C₁₉H₁₇F₅NO₅PNa⁺[M+Na]⁺:488.0657,found:488.0670。

Example 11: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclopropylmethyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with L-alanine cyclopropylmethyl ester (1.432 g, 10.0mmol) to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclopropylmethyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.26-0.30(m,2H),0.55-0.60(m,2H), 1.09-1.16(m,1H),1.49(d, J ═ 6.8Hz,3H),3.94-3.98(m,3H),4.18-4.24(m, 1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 173.13,173.05,150.23, 150.16,129.85,125.65,120.10,120.05,70.65,50.61,50.59,21.08,21.04, 9.69, 3.27; nuclear magnetic resonance³¹P NMR(160MHz,CDCl₃) Delta-1.68; high resolution mass spectrum HRMS (ESI) calcd. for C₁₉H₁₇F₅NO₅PNa⁺[M+Na]⁺:488.0657,found:488.0668。

Example 12: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine n-pentyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.591g (10.0mmol) of L-alanine n-pentyl ester to obtain the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine n-pentyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.90(t, J ═ 7.2Hz,3H),1.29-1.35(m, 4H),1.47(d, J ═ 7.2Hz,3H),1.60-1.67(m,2H),3.94-3.99(m,1H),4.13(t, J ═ 6.8Hz,2H),4.17-4.21(m,1H),7.19-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz, CDCl₃) δ 173.08,172.99,150.22,150.15,129.86,125.66,120.09,120.04, 66.02,50.57,50.55,28.20,27.91,22.25,21.06,21.02, 13.89; nuclear magnetism³¹P NMR(160 MHz,CDCl₃) Delta-1.68; high resolution mass spectrum HRMS (ESI) calcd.for C₂₀H₂₁F₅NO₅PNa⁺[M+Na]⁺: 504.0970,found:504.0993。

Example 13: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine isoamyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with 1.591g (10.0mmol) of isoamyl L-alanine to obtain the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -isoamyl L-alanine.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.91(d, J ═ 6.8Hz,6H),1.47(d, J ═ 6.8Hz,3H),1.52-1.55(m,2H),1.64-1.71(m,2H),3.94-3.98(m,1H),4.17 (t, J ═ 6.8Hz,2H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 173.06,172.97,150.18,150.11,129.86,125.66,120.07,120.03, 64.55,50.55,50.53, 37.16,24.98,22.37,21.05, 21.01; nuclear magnetic resonance³¹P NMR(160MHz,CDCl₃) Delta-1.68; high resolution mass spectrum HRMS (ESI) calcd.for C₂₀H₂₁F₅NO₅PNa⁺[M+Na]⁺:504.0970,found: 504.0991。

Example 14: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine-3-pentyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with L-alanine 3-pentyl ester (1.591 g, 10.0mmol), to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine 3-pentyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.87(td, J ═ 7.6,3.2Hz,6H),1.48(d, J ═ 6.8Hz,3H),1.52-1.64(m,4H),3.99(dd, J ═ 11.6,9.2Hz,1H),4.08-4.18 (m,1H),5.18-5.22(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) Delta 172.92,172.83,150.19,150.12,129.86,125.66,120.10,120.05,78.77, 50.67,26.41,26.35,21.28,21.24,9.51, 9.43; nuclear magnetic resonance³¹P NMR (160MHz, CDCl3): delta-1.62; high resolution mass spectrum HRMS (ESI) calcd.for C₂₀H₂₁F₅NO₅PNa⁺[M+Na]⁺:504.0970,found: 504.0986。

Example 15: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine neopentyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.591g (10.0mmol) of L-alanine neopentyl ester to obtain the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine neopentyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.94(s,9H),1.49(d, J ═ 7.2Hz,3H), 3.85(dd, J ═ 26.4,10.4Hz,3H),3.96-4.01(m,1H),4.20-4.26(m,1H), 7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) Delta 173.10,173.01,150.18, 150.11,129.87,125.68,120.10,120.05,75.03,50.56,50.54,31.44,26.29, 21.20 and 21.15; nuclear magnetism³¹P NMR(160MHz,CDCl₃) Delta-1.62; high resolution mass spectrum HRMS (ESI) calcd. for C₂₀H₂₁F₅NO₅PNa⁺[M+Na]⁺:504.0970,found:504.0978。

Example 16: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclopentyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.571g (10.0mmol) of L-alanine cyclopentyl ester to obtain the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclopentyl ester.

Nuclear magnetism of it¹H NMR(400MHz,CDCl₃):δ1.44(d,J＝6.8Hz,3H),1.59-1.72(m, 6H),1.83-1.89(m,2H),3.99(dd,J＝12.0,9.6Hz,1H),4.14-4.24(m,1H), 4.77-4.83(m,1H),7.20-7.38(m,5H)；¹³C NMR(100MHz,CDCl₃):δ172.75,172.66, 150.20,150.13,129.85,125.65,125.64,120.07,120.03,78.86,50.59,50.58, 32.63,32.51,23.64,23.64,20.97,20.93；³¹P NMR(160MHz,CDCl₃) Delta-1.60; high resolution mass spectrum HRMS (ESI) calcd.for C₂₀H₁₉F₅NO₅PNa⁺[M+Na]⁺:502.0813,found: 502.0748。

Example 17: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclobutyl methyl ester

The procedure is as in example 1, except that 1.571g (10.0mmol) of L-cyclobutyl alanine methyl ester is substituted for L-2-ethylbutyl alanine in the starting material to obtain [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-cyclobutyl alanine methyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 1.44(d, J ═ 6.8Hz,3H),1.72-1.79(m, 2H),1.84-1.96(m,2H),2.01-2.09(m,2H),3.96(dd, J ═ 12.0,9.6Hz,1H), 4.12(dd, J ═ 6.8,2.0Hz,2H),4.14-4.23(m,1H),4.77-4.83(m,1H),7.20-7.38 (m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 173.19,173.10,150.19,150.12,129.86, 125.66,120.08,120.04,69.51,50.53,33.90,24.59,21.16,21.11, 18.37; nuclear magnetism³¹P NMR(160MHz,CDCl₃) Delta-1.66; high resolution mass spectrum HRMS (ESI) calcd.for C₂₀H₁₉F₅NO₅PNa⁺[M+Na]⁺:502.0813,found:502.0883。

Example 18: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine n-hexyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.733g (10.0mmol) of L-alanine n-hexyl ester to obtain the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine n-hexyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.89(t, J ═ 6.8Hz,3H),1.29-1.37(m, 6H),1.47(d, J ═ 6.8Hz,3H),1.59-1.66(m,2H),3.96(dd, J ═ 11.6,9.2Hz,1H), 4.13(t, J ═ 6.8, Hz,2H),4.17-4.23(m,1H),7.20-7.38(m, 5H); nuclear magnetism of¹³C NMR(100MHz,CDCl₃) δ 173.07,172.99,150.19,150.12,129.85,125.67, 120.08,120.03,66.03,50.55,31.34,28.45,25.42,22.48,21.06,21.02, 13.96; nuclear magnetism of³¹P NMR(160MHz,CDCl₃) Delta-1.67; high resolution mass spectrum HRMS (ESI) calcd.for C₂₁H₂₃F₅NO₅PNa⁺[M+Na]⁺:518.1126,found:518.1099。

Example 19: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine isohexyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with 1.733g (10.0mmol) of isohexyl L-alanine to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine isohexyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.88(d, J ═ 6.8Hz,6H),1.18-1.24(m, 2H),1.47(d, J ═ 7.2Hz,3H),1.52-1.58(m,1H),1.60-1.67(m,2H),3.93-3.99 (m,1H),4.12(t, J ═ 6.8Hz,2H),4.15-4.22(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 173.08,172.99,150.22,150.15,129.86,125.67,120.09,120.05, 66.29,50.57,50.55,34.83,27.69,26.41,22.43,21.07, 21.03; nuclear magnetism³¹P NMR(160MHz,CDCl₃) Delta-1.67; high resolution mass spectrum HRMS (ESI) calcd.for C₂₁H₂₃F₅NO₅PNa⁺[M+Na]⁺:518.1126,found:518.1126。

Example 20: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclohexyl ester

The procedure is as in example 1 except that L-alanine (2-ethylbutyl) ester in the starting material is replaced with 1.712g (10.0mmol) of L-alanine cyclohexyl ester to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclohexyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃):δ1.26-1.56(m,6H),1.46(d,J＝6.8Hz, 3H),1.71-1.74(m,2H),1.81-1.85(m,2H),3.96(dd,J＝120,9.6Hz,1H), 4.13-4.19(m,1H),4.78-4.83(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz, CDCl₃) δ 172.44,172.35,150.21,150.13,129.85,125.65,120.09,120.04, 74.37,50.65,31.36,31.32,25.23,23.54,21.12, 21.08; nuclear magnetism³¹P NMR(160MHz, CDCl₃) Delta-1.58; high resolution mass spectrum HRMS (ESI) calcd.for C₂₁H₂₁F₅NO₅PNa⁺[M+Na]⁺: 516.0970,found:516.0938。

Example 21: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclopentylmethyl ester

The procedure is as in example 1 except that the starting material L-alanine (2-ethylbutyl) ester is replaced with L-alanine cyclopentylmethyl ester 1.712g (10.0mmol) to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclopentylmethyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 1.82-1.27(m,2H),1.47(d, J ═ 7.2Hz,3H), 1.56-1.63(m,4H),1.70-1.78(m,2H),2.17-2.24(m,1H),3.94-4.01(m, 1H),4.03(dd, J ═ 7.2,2.8Hz,2H),4.14-4.24(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 173.11,173.02,150.19,150.12,129.86,125.65, 120.08,120.03,69.74,50.56,50.54,38.44,29.23,29.20,25.27,21.12, 21.07; nuclear magnetism³¹P NMR(160MHz,CDCl₃) Delta-1.65; high resolution mass spectrum HRMS (ESI) calcd.for C₂₁H₂₁F₅NO₅PNa⁺[M+Na]⁺:516.0970,found:516.0961。

Example 22: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclohexylmethyl ester

The procedure is as in example 1, except that the starting material L-alanine (2-ethylbutyl) ester is replaced with 1.853g (10.0mmol) of L-alanine cyclohexylmethyl ester to give the title product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine cyclopentylmethyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 0.91-1.01(m,2H),1.13-1.28(m,3H),1.48 (d, J ═ 7.2Hz,3H),1.60-1.74(m,6H),3.94-4.01(m,1H),3.95(d, J ═ 6.4Hz,2H), 3.97-4.00(m,1H),7.20-7.38(m, 5H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 173.09,173.00,150.18,150.11,129.86,125.67,120.09,120.04,70.91, 50.54,37.03,29.51,29.49,26.26,25.58,21.13, 21.09; nuclear magnetism³¹P NMR(160MHz, CDCl₃) Delta-1.66; high resolution mass spectrum HRMS (ESI) calcd.for C₂₂H₂₃F₅NO₅PNa⁺[M+Na]⁺: 530.1126,found:530.1137。

Example 23: preparation of [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine benzyl ester

The procedure is as in example 1, except that the starting material L-alanine (2-ethylbutyl) ester is replaced with 1.792g (10.0mmol) of L-alanine benzyl ester to give the desired product [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl ] -L-alanine benzyl ester.

Nuclear magnetism of¹H NMR(400MHz,CDCl₃) δ 1.48(d, J ═ 7.2Hz,3H),1.60-1.74(m,6H),3.94-4.01(m,1H),3.95 (dd, J ═ 12.09.6 Hz,1H),4.20-4.29(m,1H), 5.17(s,2H),7.19-7.26(m,3H),7.32-7.36(m, 7H); nuclear magnetism¹³C NMR(100MHz,CDCl₃) δ 172.85,172.76,150.14,150.07,135.05,129.85,128.67,128.59,128.28, 125.67,120.06,120.01,67.51,50.57,50.49,20.93, 20.88; nuclear magnetism³¹P NMR(160 MHz,CDCl₃) Delta-1.78; high resolution mass spectrum HRMS (ESI) calcd.for C₂₂H₁₇F₅NO₅PNa⁺[M+Na]⁺: 524.0657,found:524.0742。

Example 24: preparation of Remdesivir (Remdesivir, GS-5734) based on the synthetic intermediate of the present invention

GS-4415240.291 g (1.0mmol) and

[ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl]0.676g (1.5 mmol) of isopropyl-L-alaninate, 5.0mL of DMF, N₂Protecting, and magnetically stirring at room temperature. 1.5mL (1.0M, 1.5mmol) of THF solution was added to the reaction mixture, and the mixture was reacted overnight at room temperature, 20.0mL of ethyl acetate and saturated NaHCO were added to the reaction mixture₃Solution wash (15.0 mL. times.3), saturated NaCl solution wash (15.0 mL. times.1), anhydrous Na₂SO₄Drying, filtering, evaporating the solvent, and separating by silica gel column chromatography under the gradient elution condition of methanol/dichloromethane (0/100-10/90) to obtain the target product of 0.199g of Reidesciclovir and a white solid with the yield of 33%.

Nuclear magnetism of¹H NMR(600MHz,CDCl₃) δ 0.80(t, J ═ 7.8Hz,6H),1.21-1.27(m, 7H),1.40-1.44(m,1H),3.79-3.85(m,1H),3.86-3.89(m,1H),3.94-3.97(m, 2H),4.08-4.11(m,1H),4.23-4.28(m,2H),4.65(t, J ═ 5.4,1H),5.38(d, J ═ 6.0Hz,1H),6.05(dd, J ═ 12.6,10.2Hz,1H),6.34(d, J ═ 6.6Hz,1H), 6.87(dd, J ═ 39.0,4.2, 2H),7.16-7.20(m, 3.20H), 7.93 (t, 7.93H), 7.93 (m,7H), 3.93 (m, 2H); nuclear magnetism¹³C NMR(125MHz,CDCl₃) δ 173.65,173.62,156.05,151.20, 151.15,148.37,130.04,124.96,123.96,120.53,120.50,117.36,117.01,110.79, 101.28,82.55,82.50,79.46,74.49,70.37,66.56,65.90,65.87,50.18,23.01, 22.98,20.25,20.20,11.23, 11.19; nuclear magnetism³¹P NMR(240MHz,CDCl₃) Delta 3.69; high resolution mass spectrum HRMS (ESI) calcd.for C₂₇H₃₆N₆O₈P⁺[M+H]⁺:603.2327,found:603.2322。

Example 25: preparation of Sofosbuvir (Sofosbuvir) based on the intermediate synthesized by the invention

To a reaction flask equipped with a magnetic stirrer was added PSI-62060.260 g (1.0mmol) and [ (2s) - (2,3,4,5, 6-pentafluoro-phenoxy) -phenoxy-phosphoryl]0.544g (1.2 mmol) of isopropyl-L-alaninate, 2.5mL of pyridine, N₂Protection and magnetic stirring. Cooling the reaction solution to 0 ℃, and adding Me₂0.56mL (0.9M, 0.5mmol) of AlCl n-hexane solution is gradually heated to room temperature, reaction is continued for 24H, and H is added into the reaction solution₂0.5mL of O is quenched, the solvent is evaporated to dryness, and the product is separated by silica gel column chromatography under the condition of gradient elution of methanol/dichloromethane (0/100-10/90), so that 0.381g of the target product sofosbuvir is obtained, and the yield is 72 percent.

Nuclear magnetism of¹H NMR(600MHz,CDCl₃) δ 1.15(d, J ═ 6.0,6H),1.23-1.27(m,6H), 3.79-3.84(m,2H),4.00-4.02(m,1H),4.24-4.25(m,1H),4.36-4.38(m,1H), 4.84-4.88(m,1H),5.55(d, J ═ 7.8Hz,1H),5.86(s,1H),6.04-6.08(m,2H), 7.09(t, J ═ 7.19,1H),7.22(d, J ═ 8.4Hz,2H),7.38(t, J ═ 7.8Hz,2H), 7.57(d, J ═ 6.0, 1H),11.53(s, 1H); nuclear magnetism¹³C NMR(125MHz,CDCl₃) δ 173.07, 173.04,163.20,151.17,151.12,150.89,130.12,125.05,120.53,120.50,102.72, 101.35,100.14,68.45,50.24,21.87,21.84,20.27,20.23,17.08, 16.92; nuclear magnetism³¹P NMR(240MHz,CDCl₃) Delta 3.80; high resolution mass spectrum HRMS (ESI) calcd.for C₂₂H₃₂FN₃O₉P⁺ [M+H]⁺:530.1698,found:530.1688。

In the description of the specification, reference to the description of "one embodiment," "some embodiments," "an example," "a specific example," or "some examples" or the like means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.

The present invention is illustrated in detail by the examples described above, but the present invention is not limited to the details described above, i.e., it is not intended that the present invention be implemented by relying on the details described above. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims

1. A method for synthesizing an aryloxyphosphorylated amino acid ester represented by the formula (VI), which comprises the following steps in this order:

in the above formulae (II) to (VI),

R³each independently selected from halogen, nitro, C1-6 alkyl, C1-6 alkyl halide, C1-6 alkoxy halide, saturated or partially unsaturated C3-6 cycloalkyl, saturated or partially unsaturated C6-10 heterocyclyl, C6-10 aryl, C6-12 aralkyl, preferably selected from halogen, C1-6 alkyl, C1-6 alkoxy, nitro, C1-6 alkyl halide, further preferably selected from nitro, halogen;

X represents a halogen, preferably Cl or Br.

2. The method according to claim 1, further comprising the following process steps S0 and/or S3:

in the formula, m and R¹X has the same meaning as that expressed in claim 1,

3. The method of claim 1,

in the step S1, the method includes the steps of:

the solvent is dry diethyl ether or THF or CH₂Cl₂Or CH₃CN，

The organic base in the step of S1 is an organic base selected from triethylamine, diisopropylethylamine, triisopropylamine, triallylamine, diallylethylamine, pyridine, 1-methylimidazole, 4-dimethylaminopyridine, 2, 6-dimethylpyridine, 2, 4-dimethylpyridine, preferably 1-methylimidazole.

4. The method of claim 3,

in the step S2, the method includes the steps of:

5. The method of claim 2,

the step S0 includes the steps of:

adding 2-6 equivalents of alkali into a solution of 1H-1,2, 4-triazole at-20-60 ℃, uniformly stirring, dropwise adding 1-1.5 equivalents of a compound shown in the formula (I) into a reaction solution at the temperature, reacting for 0.5-6H, and filtering to obtain a solution containing a compound shown in the formula (II)Solutions of the compounds in dry ether or THF or CH as solvent₂Cl₂Or CH₃CN, wherein the base is triethylamine, diisopropylethylamine, triisopropylamine, triallylamine, diallyl ethylamine or pyridine.

6. The method of claim 1,

in the step S1, the compound represented by the formula (II) and the compound represented by the formula (III) are mixed at 0 to 4 ℃, the mixture is slowly warmed to room temperature to continue the reaction, and in the step S0, the reaction is carried out at 20 to 25 ℃.

7. The method according to claim 5, wherein the reactions are continuously carried out by sequentially carrying out the steps S0, S1 and S2, and the reaction solution after each step is used as it is for the reaction in the next step,

8. The method according to claim 1, wherein the aryloxyphosphorylated amino acid ester compound represented by (VI) is one of the following compounds,

9. the method of claim 5,

in the step S3, the n-solvent is selected from the group consisting of ethyl acetate, isopropyl acetate, butyl acetate, isopropyl ether, diethyl ether, methyl t-butyl ether, THF, CH₂Cl₂2-methyltetrahydrofuran or 1, 4-dioxane, in a ratio of the n-solvent to the solid of the compound of formula (VI) obtained in the step S2: adding 0.2-5g of a solid of the compound of formula (VI) to 1mL of a normal solvent, wherein the anti-solvent is selected from normal pentane, normal hexane, n-hexane,One or more of n-heptane, cyclopentane, cyclohexane, cycloheptane and petroleum ether is mixed with a solvent in any proportion, the mixture is kept stand at the temperature of-10 to 40 ℃ to precipitate a solid, and the proportion of the anti-solvent to the n-solvent is 2 to 10: 1.

10. the method according to claim 9, wherein ethyl acetate is used as a normal solvent, the anti-solvent is n-hexane or petroleum ether, the ratio of the anti-solvent to the normal solvent is 3.5-4.5: 1, and a solid is separated out by standing at the temperature of 4-25 ℃.