WO2017220028A1 - Liver delivery-based antiviral precursor drug nucleoside cyclophosphate compound and use thereof - Google Patents

Abstract

Provided are an antiviral precursor drug nucleoside cyclophosphate compound based on liver specific delivery (LSD) technology and a use thereof, and in particular, provided are a compound of formula II and an isomer, pharmaceutically acceptable salt, hydrate, and solvate thereof, and the corresponding pharmaceutical composition. Also provided are the use of the compounds alone or in combination with other antiviral drugs in the treatment of hepatitis B virus (HBV), hepatitis D virus (HDV) and human immunodeficiency virus (HIV) and diseases caused by same.

Description

Liver delivery antiviral prodrug nucleoside cyclic phosphate compound and application thereof Technical field

The present invention relates to an antiviral prodrug based on liver specific delivery (LSD), a preparation and application of a nucleoside cyclic phosphate compound, or an optical isomer, a hydrate thereof, a solvent Compounds, pharmaceutically acceptable salts, and pharmaceutical compositions.

Background technique

Viruses such as hepatitis B virus (HBV), hepatitis D virus (HDV), and human immunodeficiency virus (HIV) pose a serious threat to human health. Taking hepatitis B virus as an example, hepatitis B virus (hepatitis B) is a disease caused by hepatitis B virus, which is mainly caused by inflammatory lesions of the liver and can cause damage to multiple organs. According to the World Health Organization (WTO) survey, it is estimated that 240 million people worldwide are chronic hepatitis B infected. An estimated 780,000 people die each year from hepatitis B infection, of which 650,000 die from chronic hepatitis B. Cirrhosis and liver cancer, 130,000 people die of acute hepatitis B infection, is an important issue in global health.

Anti-hepatitis B virus drugs, the main one is nucleotides, such as: adefovir dipivoxil, tenofovir (TDF), tenofovir alafenamide (TAF), entecavir, pull Mivudine, telbivudine, etc., its mechanism of action is to activate into a triphosphate metabolite in the cell, inhibit the DNA or RNA polymerase activity of the virus, prevent DNA or RNA synthesis, and inhibit the replication of the virus.

Some nucleotide compounds, such as adefovir, tenofovir, etc., are highly electronegative at physiological pH. Therefore, oral administration, poor transmembrane ability, low bioavailability; at the same time, increased toxic side effects of the gastrointestinal tract and kidney. However, esterification and modification to form ester prodrugs, such as adefovir dipivoxil, tenofovir, etc., can improve bioavailability and tissue distribution. However, the ester hydrolase is widely distributed in the body, and most of the drugs are hydrolyzed into negatively charged bioactive components (adefovir, tenofovir, etc.) before the drug reaches the liver cells, and the component does not easily enter the liver cells. The proximal tubule that is actively transported to the kidney is prone to nephrotoxicity.

The cyclic phosphate (4-aryl-2-oxo-1,3,2-dioxaphosphane) precursor structure has good liver-specific delivery properties, and the mechanism is very clear, as shown in Figure 1. As shown, the 4-aryl substitution position is specifically catalyzed by CYP3A in the cytochrome P450 isozyme family in hepatocytes to form a hydroxyl group, which is then opened to form a negatively charged phosphate intermediate, which is not readily present through the cell membrane. In the cell, under the catalysis of phosphodiesterase, after hydrolysis, β-elimination reaction, a nucleoside monophosphate compound is formed, and a nucleotide triphosphate compound is continuously produced under the action of a nucleotide kinase, and at the same time, The metabolic by-product aryl vinyl ketone can be removed by a 1,4-addition reaction with abundant antioxidant and free radical glutathione in hepatocytes, and no side effects of the addition product have been reported.

Adefovir is used as the active ingredient, through the modification of the substituents on the aryl group, monosubstituted, disubstituted, different substituents and other dozens of combined compounds, and finally found the inter-chloro-substituted aromatic ring, Pradefovir, under the action of CYP3A enzyme The rate of metabolism to adefovir is the highest, nearly five times that of 3,5-dichloroaryl (US200707214668 B2).

However, there is currently no virus-inhibiting compound with high activity, strong liver-specific delivery, and low toxic side effects. Therefore, there is an urgent need in the art to develop novel virus-inhibiting compounds having high activity, high liver-specific delivery, and low toxic side effects.

Summary of the invention

The present invention synthesizes a cyclic phosphate of an antiviral nucleotide drug, and then further modifies its aromatic ring substituent to obtain a class of prodrugs having a liver-specific delivery (liver delivery) effect, thereby making the therapeutic effect more effective. High, less toxic side effects.

In a first aspect of the invention, there is provided a compound of formula II, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof:

among them:

R _{1 is} selected from hydrogen, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1- a C6 alkylamino group; wherein said substitution has one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, nitro, hydroxy, amino, cyano;

m is 0, 1, 2, 3, 4, 5;

Each R _{2 is} independently selected from halogen, nitro, hydroxy, amino, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C6 alkane Oxy, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 carboxyl, substituted or unsubstituted C1-C6 ester, substituted or unsubstituted C2-C6 alkanoyl, substituted or not Substituted C2-C6 alkanoamide group;

Wherein said substitution has one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, nitro, hydroxy, amino, cyano;

In Formula II, each chiral center is an R type or an S type.

In another preferred embodiment, in the phosphate ring structure, P2 and its 4-aryl group are cis, and P2 is R and C4 is S.

In another preferred embodiment, said R _{1 is} selected from the group consisting of H, C1-C3 alkyl, and cyclopropyl.

In another preferred embodiment, said R _{1 is} selected from the group consisting of H, methyl, and cyclopropyl.

In another preferred embodiment, the

From the following group:

In another preferred embodiment, the compound is selected from the group consisting of:

In another preferred embodiment, the compound of formula II is a compound of formula II-a.

In another preferred embodiment, the compound has the structure shown below:

Wherein R ₃ and R ₅ are halogen;

n is 0, 1, 2, 3.

In another preferred embodiment, the compound is a compound of formula I-a and formula III-a.

In another preferred embodiment, R ₃ is halogen, R ₅ is F, Br, or I, and R ₃ ≠ R ₅ .

In another preferred embodiment, R ₃ is Cl and R ₅ is F.

In another preferred embodiment, the compound is selected from the group consisting of:

In another preferred embodiment, the salt of the compound of Formula I, Formula II and Formula III is a pharmaceutically acceptable salt of a compound of Formula I, Formula II and Formula III with an inorganic or organic acid. Or a salt of the compound of the formula I, formula II and formula III is a pharmaceutically acceptable salt formed by reacting a compound of formula I, formula II and formula III with a base. The compound of the formula I, the formula II and the formula III or a salt thereof is an amorphous substance or a crystal.

According to a second aspect of the invention, there is provided a pharmaceutical composition comprising a therapeutically effective amount of a compound as described in the first aspect of the invention, or an optical isomer thereof, a pharmaceutically acceptable salt thereof a hydrate or solvate; and a pharmaceutically acceptable adjuvant, diluent or carrier.

A third aspect of the invention provides a use of a compound according to the first aspect of the invention, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, or as in the second aspect of the invention Use of the pharmaceutical composition for the preparation of a pharmaceutical composition for treating and/or preventing acute or chronic diseases associated with hepatitis B virus (HBV), hepatitis D virus (HDV) or human immunodeficiency virus (HIV) infection .

In another preferred embodiment, the acute or chronic disease associated with hepatitis B virus (HBV), hepatitis D virus (HDV) or human immunodeficiency virus (HIV) infection is selected from the group consisting of hepatitis B, hepatitis D or AIDS.

According to a fourth aspect of the invention, there is provided a process for the preparation of a compound of formula II according to the first aspect of the invention, the method comprising the steps of:

i. Condensation of a compound of formula Va and a compound of formula Vc in an inert solvent to provide a compound of formula II.

In another preferred embodiment, in step i, the reaction is carried out in the presence of a condensing agent.

In another preferred embodiment, the condensing agent is selected from the group consisting of dicyclohexylcarbodiimide (DCC).

In another preferred embodiment, the condensation reaction is carried out at 60 to 100 ° C (about 80 ° C).

In another preferred embodiment, the condensation reaction has a reaction time of from 1 to 72 hours, preferably from 3 to 48 hours, more preferably from 6 to 24 hours.

In another preferred embodiment, the inert solvent is selected from the group consisting of N,N-dimethylformamide, pyridine, or a combination thereof; preferably 20:1 of N,N-dimethylformamide and pyridine A mixed solvent of 1:5 (v/v) (preferably 10:1 to 1:2 (v/v)).

In another preferred embodiment, the compound of formula Vc (preferably a chiral 1,3-propanediol derivative) is prepared by the following method:

Ii. In an inert solvent (such as DMF) in HCOOH, Et ₃ N and (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethylenediamine (dichloro) (pair A In the presence of cumene (II), at 40-80 ° C

Carry out a reduction reaction (such as 1-5h) to get

Iii. In amphoteric solvents (such as EtOH), with a reducing agent (such as NaBH4) and

Reaction (eg 1-5h) to get

In another preferred embodiment, the

It is prepared by any one of methods 1-3 selected from the group consisting of:

method 1

i. In an amphoteric solvent (such as EtOH), use SOCl ₂ with

Reaction, get

Compound

Ii. in an inert solvent such as THF, in the presence of a base such as LiHMDS, with ethyl acetate and

React at -60 ~ -20 ° C (such as 10 ~ 30min), get

Compound

Method 2

i. in an inert solvent (such as DCM) in the presence of SnCl ₂ at room temperature

versus

Reaction, get

Compound

Method 3

i. In an inert solvent (such as THF), in the presence of a base (such as potassium t-butoxide),

Reacting at reflux temperature overnight,

Compound.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the technical features specifically described in the following (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

DRAWINGS

Figure 1 shows a schematic diagram of the mechanism of action of liver delivery compounds.

Figure 2 shows the ratio of each liver-delivering compound (racemate) metabolized to an active molecule by the action of the CYP3A4 enzyme. The compound names are shown in Table 1. The active metabolic molecule corresponding to all compounds is PMPA.

Figure 3 shows the ratio of liver delivery compounds (S configuration cis) metabolized to active molecules by the action of the CYP3A4 enzyme. The compound names are shown in Table 1. Wherein, the compound 6-cis corresponding active metabolic molecule is PMPA, and the active metabolic molecule corresponding to compound 9 (Pradefovir) is PMEA.

Figure 4 is a graph showing the concentration-time columnar distribution of the metabolically released active molecule PMPA in plasma, liver and kidney after intragastric administration of 30 mg/kg of PA1010 in rats. The compound names are shown in Table 1.

Figure 5 is a graph showing the concentration-time columnar distribution of the metabolically released active molecule PMPA in plasma, liver and kidney after intragastric administration of 30 mg/kg of PA1007 in rats. The compound names are shown in Table 1.

Figure 6 is a graph showing the concentration-time columnar distribution of the metabolically released active molecule PMPA in plasma, liver and kidney after intragastric administration of 30 mg/kg of TAF in rats.

Figure 7 is a graph showing the concentration-time columnar distribution of the metabolically released active molecule PMPA in plasma, liver and kidney after intragastric administration of 30 mg/kg of TDF in rats.

Note:

PMPA: (R)-9-(2-phosphomethoxypropyl)-adenine

PMEA: 9-[2-(phosphonomethoxy)ethyl]adenine

S configuration cis: If there is no special expression, it means that C4 is the S configuration in the phosphate ring structure, and P2 and its 4-position aromatic group are cis.

detailed description

After long-term and intensive research, the present inventors have discovered for the first time through a screening study of a large number of compounds: a class of compounds of formula I and formula III having a specific structure (for example, the 3 and 5 positions of the benzene ring moiety are different halogens, or The 2- and 5-positions of the phenyl ring moiety are different halogens, surprisingly having very excellent antiviral activity, significantly improved liver-specific delivery (liver delivery), and significantly reduced toxic side effects. Based on the above findings, the inventors completed the present invention.

the term

As used herein, the term "C1-C6 alkyl" refers to a straight or branched alkyl group having from 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, A sec-butyl group, a tert-butyl group, or the like.

The term "C2-C6 alkanoyl" as used herein refers to a substituent of the structure "linear or branched alkyl-carbonyl having 1 to 6 carbon atoms", such as acetyl, propionyl, butyryl, or Similar group.

As used herein, the term "C1-C6 alkylamino" refers to a substituent of the form "linear or branched alkyl-amino" having 1 to 6 carbon atoms, such as methylamino, dimethylamino. , ethylamino, propylamino, diethylamino, or the like.

The term "halogen" refers to F, Cl, Br and I.

In the present invention, the terms "containing", "comprising" or "including" mean that the various ingredients may be used together in the mixture or composition of the present invention. Therefore, the terms "consisting essentially of" and "consisting of" are encompassed by the term "contains."

In the present invention, the term "pharmaceutically acceptable" ingredient means a substance which is suitable for use in humans and/or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reaction), that is, a reasonable benefit/risk ratio.

In the present invention, the term "effective amount" means an amount of a therapeutic agent that treats, alleviates or prevents a target disease or condition, or an amount that exhibits a detectable therapeutic or prophylactic effect. The precise effective amount for a subject will depend on the size and health of the subject, the nature and extent of the condition, and the combination of therapeutic and/or therapeutic agents selected for administration. Therefore, it is useless to specify an accurate effective amount in advance. However, for a given condition, routine experimentation can be used to determine the effective amount that the clinician can determine.

As used herein, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, nitrate Base, hydroxyl, amino, cyano group.

Unless otherwise stated, all compounds appearing in the present invention are meant to include all possible optical isomers, such as a single chiral compound, or a mixture of various chiral compounds (i.e., racemates). Among all the compounds of the present invention, each of the chiral carbon atoms may be optionally in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

The term "compound of the invention" as used herein refers to a compound of formula II. The term also encompasses various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of Formula II.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of the compound of the invention formed with an acid or base suitable for use as a medicament. Pharmaceutically acceptable salts include inorganic and organic salts. A preferred class of salts are the salts of the compounds of the invention with acids. Suitable The acid forming the salt includes, but is not limited to, mineral acid such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, Organic acids such as maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzoic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

Some of the compounds in the present invention may be crystallized or recrystallized with water or various organic solvents, in which case various solvates may be formed. The solvates of the present invention include stoichiometric solvates such as hydrates and the like, as well as compounds containing variable amounts of water formed upon preparation by low pressure sublimation drying.

It will be understood that various thermodynamically stable isomers may be present after preparation of the compounds of the invention, such as tautomers, conformers, meso compounds, and optical isomers having enantiomeric or diastereomeric relationships. The above-described modifications will become apparent to those skilled in the art upon reading the disclosure of the present invention.

Compounds of formula I and formula III and preparation thereof

In order to provide a highly potent, low toxic prodrug capable of releasing antiviral nucleotides in hepatocytes by a liver-specific delivery mechanism, the inventors have prepared preferred compounds of formula II and formula III compound:

among them:

R _{1 is} selected from hydrogen, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1- a C6 alkylamino group; wherein said substitution has one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, nitro, hydroxy, amino, cyano;

m is 0, 1, 2, 3, 4, 5;

Each R _{2 is} independently selected from halogen, nitro, hydroxy, amino, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C6 alkane Oxy, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 carboxyl, substituted or unsubstituted C1-C6 ester, substituted or unsubstituted C2-C6 alkanoyl, substituted or not Substituted C2-C6 alkanoamide group;

Wherein said substitution has one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, nitro, hydroxy, amino, cyano;

In Formula II, each chiral center is an R type or an S type.

In a preferred embodiment, the

From the following group:

A preferred class of compounds of formula II have the structure of formula I below:

In another preferred embodiment, the compound of formula I is a compound of formula I-a.

In another preferred embodiment, the P2 and the 4-position aromatic groups in the phosphate ring structure are cis to each other, and P2 is an R type, and C4 is an S type.

In another preferred embodiment, said R _{1 is} selected from the group consisting of H, C1-C3 alkyl, and cyclopropyl.

In another preferred embodiment, said R _{1 is} selected from the group consisting of H, methyl, and cyclopropyl.

In a more preferred case, R ₃ is Cl and R ₅ is F; or R ₃ is Cl and R ₅ is Br; or R ₃ is Cl and R ₅ is Cl.

In another preferred embodiment, the optical isomers include tautomers, cis and trans isomers, conformational isomers, meso compounds, and optical isomers having enantiomeric or diastereomeric relationships. .

In another preferred embodiment, the compound is selected from the group consisting of:

Another preferred class of compounds of formula II has the structure shown in formula III below:

Among them, the definition of each group is as shown above.

In another preferred embodiment, the compound of formula III is a compound of formula III-a.

In another preferred embodiment, the P2 and the aromatic group at the 4-position in the phosphate ring structure are cis to each other, and P2 is an R type, and C4 is an S type.

In another preferred embodiment, said R _{1 is} selected from the group consisting of H, C1-C3 alkyl, and cyclopropyl.

In another preferred embodiment, said R _{1 is} selected from the group consisting of H, methyl, and cyclopropyl.

In a more preferred case, R ₃ is Cl and R ₅ is F; or R ₃ is Cl and R ₅ is Br; or R ₃ is Cl and R ₅ is Cl.

In another preferred embodiment, the optical isomers include tautomers, cis and trans isomers, conformational isomers, meso compounds, and optical isomers having enantiomeric or diastereomeric relationships. .

In another preferred embodiment, the compound is selected from the group consisting of:

Method for preparing a compound (taking a compound of formula III as an example):

In a solution of N,N-dimethylformamide and pyridine (5:1), a monophosphate derivative Va, a 1,3-propanediol derivative Vd and dicyclohexylcarbodiimide are added, and the temperature is raised to about 80 ° C. After the reaction was completed, the reaction was completed, and the reaction mixture was evaporated to dryness. a compound of formula II.

Here, each of the reactants may be commercially available or may be prepared by a conventional method in the art using commercially available raw materials.

In a preferred embodiment of the invention, the 1,3-propanediol derivative Vd (preferably a chiral 1,3-propanediol derivative) is prepared by the following method:

i. In an inert solvent (such as DMF) in HCOOH, Et ₃ N and (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethylenediamine (dichloro) (pair A In the presence of cumene (II), at 40-80 ° C

Carry out a reduction reaction (such as 1-5h) to get

Ii. In an amphoteric solvent (such as EtOH), a reducing agent (such as NaBH ₄ ) and

Reaction (eg 1-5h) to get

In another preferred embodiment, the

It is prepared by any one of methods 1-3 selected from the group consisting of:

method 1

i. In an amphoteric solvent (such as EtOH), use SOCl ₂ with

Reaction, get

Compound

Ii. in an inert solvent such as THF, in the presence of a base such as LiHMDS, with ethyl acetate and

React at -60 ~ -20 ° C (such as 10 ~ 30min), get

Compound

Method 2

i. in an inert solvent (such as DCM) in the presence of SnCl ₂ at room temperature

versus

Reaction, get

Compound

Method 3

i. In an inert solvent (such as THF), in the presence of a base (such as potassium t-butoxide),

Reacting at reflux temperature overnight,

Compound.

It should be understood that the above preparation method is only taking the compound of the formula III as an example, and those skilled in the art can conveniently replace the corresponding starting materials to prepare other compounds of the formula I and formula II.

Pharmaceutical composition and method of administration

Since the compound of the present invention has excellent inhibitory activity against hepatitis B virus (hepatitis B virus), the compound of the present invention and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and The pharmaceutical composition of the present invention as a main active ingredient can be used for the treatment, prevention, and alleviation of diseases caused by hepatitis B virus. According to the prior art, the compounds of the present invention are useful for the treatment of diseases caused by infections such as HBV, HDV and HIV.

The pharmaceutical compositions of the present invention comprise a safe or effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. By "safe and effective amount" it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 0.1 to 1000 mg of the compound/agent of the invention, more preferably from 0.5 to 500 mg of the compound/agent of the invention. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By "compatibility" it is meant herein that the components of the composition are capable of intermingling with the compounds of the invention and with each other without significantly reducing the efficacy of the compound. Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid). , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as

), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration. A particularly preferred mode of administration is oral.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, hard Calcium citrate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.

In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, and flavoring agents.

In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.

Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants. The active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.

The compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight, The dose to be administered is usually 0.2 to 1000 mg, preferably 0.5 to 500 mg. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.

The main advantages of the invention include:

(1) Liver-specific delivery technology (liver delivery) is strong, and compounds can only be specifically catalyzed by CYP3A in the cytochrome P450 isozyme family in hepatocytes to produce active molecules with high negative charge, no It is easy to excrete from the liver, so it has a higher concentration in the liver and achieves a specific delivery effect.

(2) High activity, because of the specific delivery of the liver, more drugs are present in the liver, and the antiviral activity can be greatly improved.

(3) low side effects: the same dose of prodrug, the amount of active molecules metabolized outside the liver is very small, so the kidney toxicity and bone toxicity performance are greatly reduced.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

Example 1 (2R)-9-{2-[(4S)-4-(3-chloro-2-fluorophenyl)-2-oxo-1,3,2-dioxaphospholane Preparation of 2-yl]methoxypropyl}adenine

(R)-9-[2-(phosphonomethoxy)propyl]-adenine (84 mg, 0.294 mmol) was dissolved in N,N-dimethylformamide (15 mL) and pyridine (3 mL) Dicyclohexylcarbodiimide (182 mg, 0.882 mmol) and (S)-3-(3-chloro-2-fluorophenyl)-1,3-propanediol (60 mg, 0.294 mmol) were added and the reaction mixture was heated to 80 The reaction was carried out for 16 h, and the reaction was completed. The reaction mixture was evaporated to dryness crystals eluted eluted eluted eluted eluted eluted eluted eluted Analysis (dichloromethane:methanol = 20:1 to 10:1) gave white solid, yield: 41%, _Rf = 0.4 (dichloromethane:methanol = 10:1), ¹ H NMR (400 MHz, DMSO -d ₆ ) δ: 8.071-8.227 (m, 2H), 7.202-7.661 (m, 5H), 5.837-5.961 (m, 1H), 4.549-4.603 (m, 1H), 3.935-4.356 (m, 6H) , 1.914-2.119 (m, 2H), 1.105-1.198 (m, 3H) ppm.

Example 2(2R)-9-{2-[(4S)-4-(3-chloro-5-fluorophenyl)-2-oxo-1,3,2-dioxaphospholane Preparation of 2-yl]methoxypropyl}adenine

Prepared in a similar manner to Example 1, (R)-9-[2-(phosphonomethoxy)propyl]-adenine (84 mg, 0.294 mmol), dicyclohexylcarbodiimide (182 mg, 0.882 mmol) and (S)-3-(3-chloro-5-fluorophenyl)-1,3-propanediol (60 mg, 0.294 mmol) were reacted to give 35 mg of white solid, yield: 26%, R _f = 0.4 (dichloromethane:methanol = 10:1), ¹ H NMR (400 MHz, DMSO-d ₆ ): δ: 8.045-8.163 (m, 2H), 7.173-7.455 (m, 5H), 5.621-5. , 1H), 4.228-4.271 (m, 1H), 4.001-4.059 (m, 6H), 1.952-2.109 (m, 2H), 1.097-1.196 (m, 3H) ppm.

Example 3(2R)-9-{2-[(4S)-4-(3-chloro-4-fluorophenyl)-2-oxo-1,3,2-dioxaphospholane Preparation of 2-yl]methoxypropyl}adenine

Prepared in a similar manner to Example 1, (R)-9-[2-(phosphonomethoxy)propyl]-adenine (118 mg, 0.412 mmol), dicyclohexylcarbodiimide (254.6 mg) , 1.236 mmol) and (S)-3-(3-chloro-4-fluorophenyl)-1,3-propanediol (84 mg, 0.412 mmol) were reacted to give 71 mg of white solid, yield: 37.8%, R _f = 0.4 (dichloromethane: methanol = 10:1), ¹ H NMR (400 MHz, DMSO-d ₆ ): δ: 8.056-8.151 (m, 2H), 7.552-7.584 (m, 1H), 7.25-7.493 ( m, 4H), 5.556-5.618 (m, 1H), 4.427-4.502 (m, 1H), 3.922-4.303 (m, 6H), 1.813-2.016 (m, 2H), 1.095-1.193 (m, 3H) ppm .

Example 4(2R)-9-{2-[(4S)-4-(5-chloro-2-fluorophenyl)-2-oxo-1,3,2-dioxaphospholane Preparation of 2-yl]methoxypropyl}adenine

Prepared in a similar manner to Example 1, (R)-9-[2-(phosphonomethoxy)propyl]-adenine (112 mg, 0.39 mmol), dicyclohexylcarbodiimide (242 mg, 1.176 mmol) and (S)-3-(5-chloro-2-fluorophenyl)-1,3-propanediol (80 mg, 0.39 mmol) were reacted to give 60 mg of white solid, yield: 38%, R _f = 0.3 (dichloromethane:methanol = 10:1), ¹ H NMR (400 MHz, DMSO-d ₆ ): δ: 8.48-8.225 (m, 2H), 7.182-7.545 (m, 5H), 5.825-5.911 (m) , 1H), 4.536-4.593 (m, 1H), 3.971-4.339 (m, 6H), 1.893-2.172 (m, 2H), 1.103-1.196 (m, 3H) ppm.

(2R)-9-{2-[(4S)-4-(5-chloro-2-fluorophenyl)-2-oxo-1,3,2-dioxaphosphinohexane-2- 4-methoxypropyl}adenine (15.66g) was passed through a chiral column to give the 4-cis (PA1010) isomer under the following conditions:

(2R)-9-{2-[(2R,4S)-4-(5-chloro-2-fluorophenyl)-2-oxo-1,3,2-dioxaphospholane- 2- yl] methoxy} propyl adenine ^{(9.69g), 1 H NMR (} 400MHz, Methanol-D4): δ: 8.19 (s, 1H), 8.14 (s, 1H), 7.43-7.34 (m, 2H), 7.19–7.11 (m, 1H), 5.86 (t, J=7.0 Hz, 1H), 4.70–4.60 (m, 1H), 4.46–4.26 (m, 3H), 4.16–4.08 (m, 1H) , 4.05 (t, J = 7.3 Hz, 2H), 2.16 - 2.08 (m, 2H), 1.30 (d, J = 6.2 Hz, 3H). ppm

Pillar: CHIRALPAK ADH

Mobile phase: ethanol: acetonitrile = 90:10 (V / V)

Wavelength: UV254nm

Temperature: 25 degrees

Example 5(2R)-9-{2-[(4S)-4-(4-Pyridinyl)-2-oxo-1,3,2-dioxaphosphinopentan-2-yl]- Preparation of oxypropyl}adenine

Prepared in a similar manner to Example 1, (R)-9-[2-(phosphonomethoxy)propyl]-adenine (112 mg, 0.39 mmol), dicyclohexylcarbodiimide (242 mg, 1.176 mmol) and (S) -3- (4- pyridinyl) -1,3-propanediol (80mg, 0.39mmol) completion of the reaction, to give 70mg as a white solid, _{yield: 39%, R f = 0.4} ( methylene chloride: Triethylamine: methanol = 10:1: 0.1), ¹ H NMR (400 MHz, DMSO-d ₆ ): δ: 8.495-8.629 (m, 1H), 8.545 (d, J = 4.8HZ, 1H), 8.070- 8.169 (m, 2H), 7.213-7.350 (m, 4H), 5.591-5.662 (m, 1H), 4.483-4.559 (m, 1H), 3.913-4.345 (m, 6H), 1.913-2.114 (m, 2H) ), 1.100-1.178 (m, 3H) ppm.

Example 6(2R)-9-{2-[(2R,4S)-4-(3-chloro-5-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorane Alkano-2-yl]methoxypropyl}adenine and (2R)-9-{2-[(2S,4S)-4-(3-chloro-5-fluorophenyl)-2-oxo Preparation of -1,3,2-dioxaphosphinohexan-2-yl]methoxypropyl}adenine

(2R)-9-{2-[(4S)-4-(3-chloro-5-fluorophenyl)-2-oxo-1,3,2-dioxaphospholane-2- The methoxypropyl}adenine (410.7 mg) was passed through a chiral column to give the diastereomer under the following conditions: (2R)-9-{2-[(2R,4S)-4 -(3-chloro-5-fluorophenyl)-2-oxo-1,3,2-dioxaphosphinopipetan-2-yl]methoxypropyl}adenine (220.2 mg), _{^{1 H NMR (400MHz, CDCl 3}} ): δ: 8.350 (S, 1H), 7.930 (S, 1H), 7.074-7.100 (m, 2H), 6.890 (d, J = 8.8Hz, 1H), 5.735 (s , 2H), 5.585 (d, J = 10.4 Hz, 1H), 4.655-4.711 (m, 1H), 4.343-4.429 (m, 2H), 4.157-4.212 (m, 1H), 3.970-4.059 (m, 2H) ), 3.801-3.860 (m, 1H), 2.014-2.108 (m, 2H), 1.316 (d, J = 6.4 Hz, 3H) ppm. and (2R)-9-{2-[(2S, 4S)- 4-(3-Chloro-5-fluorophenyl)-2-oxo-1,3,2-dioxaphosphinohexan-2-yl]methoxypropyl}adenine (166.2 mg) , ¹ H NMR (400 MHz, CDCl ₃ ): δ: 8.372 (m, 1H), 7.095 (s, 1H), 7.011-7.157 (m, 3H), 5.899 (s, 2H), 5.432 (d, J = 11.2) Hz, 1H), 3.897-4.410 (m, 7H), 2.141-2.249 (m, 1H), 1.763-1.800 (m, 1H), 1.337 (d, J = 6.4 Hz, 3H) ppm.

Pillar: CHIRALPAK ADH

Mobile phase: ethanol: acetonitrile = 90:10 (V / V)

Wavelength: UV254nm

Temperature: 25 degrees

Comparative Example 7(2R)-9-{2-[(4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphol-2-yl Preparation of methoxypropyl}adenine

Prepared in a similar manner to Example 1, (R)-9-[2-(phosphonomethoxy)propyl]-adenine (660 mg, 2.419 mmol), dicyclohexylcarbodiimide (1.5 g) , 7.257 mmol) and (S)-3-(3-chlorophenyl)-1,3-propanediol (450 mg, 2.419 mmol) were reacted to give 400 mg of white solid, yield: 38%, R _f = 0.5 (2) Methyl chloride:methanol = 10:1), ¹ H NMR (400MHz, DMSO-d ₆ ): δ: 7.872-8.296 (m, 2H), 7.211-7.269 (m, 4H), 6.019-6.077 (m, 2H) , 5.523-5.550 (m, 1H), 4.261-4.357 (m, 1H), 3.773-4.156 (m, 6H), 1.907-1.990 (m, 2H), 1.236-1.354 (m, 3H) ppm.

Comparative Example 8(2R)-9-{2-[(4S)-4-(3,5-dichlorophenyl)-2-oxo-1,3,2-dioxaphospholane- Preparation of 2-yl]methoxypropyl}adenine

Prepared in a similar manner to Example 1, (R)-9-[2-(phosphonomethoxy)propyl]-adenine (84 mg, 0.294 mmol), dicyclohexylcarbodiimide (182 mg, After completion of the reaction with (S)-3-(3,5-dichlorophenyl)-1,3-propanediol (60 mg, 0.294 mmol), 62 mg of white solid was obtained, yield: 45%, _Rf = 0.4. (Dichloromethane: methanol = 10:1), ¹ H NMR (400 MHz, CDCl ₃ ): δ: 8.346-8.378 (m, 1H), 7.09-7.939 (m, 1H), 7.351-7.363 (m, 1H) , 7.265-7.268 (m, 1H), 7.178-7.181 (m, 1H), 5.770-5.825 (m, 2H), 5.399-5.588 (m, 1H), 3.808-4.428 (m, 7H), 2.025-2.085 ( m, 2H), 1.306-1.349 (m, 3H) ppm.

Comparative Example 9(2R)-9-{2-[(2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphospholane-2 Preparation of -yl]methoxyethyl}adenine

It is prepared by the technical content disclosed in the article: J, Am, Chem, Soc, 2004, 126, 5154-5163. _{^{1 H NMR (400MHz, CDCl 3}} ): δ: 8.352 (s, 1H), 7.907 (s, 1H), 7.285-7.354 (m, 3H), 7.106 (d, J = 6.8Hz, 1H), 5.802 (s , 2H), 5.591 (d, J = 10.8 Hz, 1H), 4.624-4.681 (m, 1H), 4.443-4.468 (m, 2H), 4.235-4.321 (m, 1H), 3.899-4.031 (m, 4H) ), 1.980-2.109 (m, 2H) ppm.

Table 1 The compounds prepared in the respective examples are shown in the following table:

Note: Unless otherwise stated, PA1010 refers to PA1010-cis, PA1007 refers to PA1007-cis.

Example 11 Evaluation of in vitro CYP3A4 Enzyme Metabolism to Active Molecule

test methods:

The efficiency of metabolism into active molecules (PMPA or PMEA) by the determination of a prodrug at a concentration of 0.1 μM at a concentration of 1 mg/ml of human recombinant CYP3A4 enzyme (CYPEX) was evaluated. The enzymatic reaction was carried out in a volume of 500 μl, a 0.1 M solution of pH 7.4 in Tris-HCl buffer solution containing 5 mM magnesium chloride and 1 mM NADPH. The reaction mixture was incubated in a constant temperature shaking water bath at 37 ° C for 0, 7, 17, 30 minutes, and the reaction was stopped by adding 1.5 volumes of methanol. Centrifuge for 20 minutes at a maximum speed of 13,600 rpm using an Eppendorf bench top centrifuge. The supernatant was taken, dried by a nitrogen blower and then redissolved to mobile phase A (aqueous solution containing 5 mM ammonium acetate and 0.05% formic acid v/v). The resulting sample was obtained by LC-MS/MS (Waters, Acquity UPLC HSS) T3 column) for analysis.

Table 2 The amount of CYP3A4 enzyme metabolized into active molecules in vitro

Analysis of the results: Compounds 1, 2, 3, 4, 5, 7 and 8 are S-type racemate structures, 6 and 9 are S-cis structures, Table 2 is the 30-minute enzyme metabolism results, Figure 2 and Figure 3 are Dynamic results of enzyme metabolism. As can be seen from Table 2, in the S-form racemic structure, the ratio of the compound 2 to the active metabolic molecule PMPA was the highest, being 13.29%, and the comparative examples 7 and 8 were: 8.67% and 1.01%, respectively.

Compound 2 was resolved to give a S-configuration cis (compound 6-cis) metabolic ratio of 27.36%, which is higher than its corresponding S-form racemate (Compound 2).

As can be seen from the above results, the compound 2 of the present invention is about 50% to 1300% higher than the compounds of other structures. The activity of the present compound 2 (different halogens Cl and F at the 3 and 5 positions) was about 13 times higher than that of the compound 8 in which both the 3 and 5 positions were Cl.

The most active compound 2 was resolved to obtain the cis-form 6-cis, the resolved S-configuration cis compound and the same type of compound Pradefovir (compound 9, S configuration cis) that has entered clinical research. In comparison, the activity of the compound 6-cis of the present invention is also about 57.5% higher.

Example 12 Evaluation of Metabolism of Human Liver Microsomes into Active Molecules in Vitro

test methods:

The human liver microsomes used in this test were purchased from In Vitro Technologies (IVT), batch number SSP X008070, which is a mixed liver microsome extracted from the liver tissue of 150 donors. The batch of liver microparticles was recorded in the product description. The metabolic activity of CYP3A4 is 1.734 nmol/mg/min (the rate at which testosterone is metabolized to produce 6-beta testosterone). The test compound was synthesized by Zhejiang Plato Pharmaceutical Technology Co., Ltd., and dissolved in methanol to prepare a storage solution having a concentration of 25 mM. The enzymatic reaction was carried out in 100 μl of a reaction solution (100 mM Tris-HCl, 5 mM MgCl ₂ , pH 7.4) at a test compound concentration of 25 μM, a human liver microsome concentration of 2 mg/ml, and a reaction was initiated by adding NADPH (final concentration 2 mM). After reacting for 5 min in a constant temperature shaking water bath, 1.5 times the volume of acetonitrile was quickly added to terminate the reaction. Centrifuge for 20 minutes at a maximum speed of 13,600 rpm using an Eppendorf bench top centrifuge. The supernatant was taken, dried by a nitrogen blower, and then redissolved to Mobile Phase A (5 mM ammonium acetate and v/v 0.05% aqueous solution). The resulting sample was quantitatively analyzed by LC-MS/MS (Waters, Acquity UPLC HSS T3 column).

Table 3 Rate of test compound metabolism from human liver microsomes to PMPA or PMEA in vitro

Note: HLM is the English abbreviation for human liver microsomes.

Analysis of results: Compounds 1, 2, 3, 4 and 7 are S-type racemic structures, 4-cis, 6-cis and 9 are S-cis structures. Table 3 shows the metabolism of human liver microsomes in 5 min. The average rate of PMPA or PMEA.

As can be seen from Table 3, in the S-form racemic structure, compounds 2 and 4 were metabolized to the active metabolic molecule PMPA at the fastest rate of 46.8 pmol/min/mg HLM and 48.8 pmol/min/mg HLM, respectively.

Compound 2 was resolved to obtain the S configuration cis (compound 6-cis), which metabolized to produce PMPA at a rate of 75.8 pmol/min/mg HLM, higher than its corresponding S configuration racemate (compound) 2).

Compound 4 was resolved to obtain the S configuration cis (4-cis), and the rate of metabolization to PMPA was 170 pmol/min/mg HLM, which was higher than its corresponding S-form racemate (Compound 4). .

It can be seen from the above results that Compound 2 and Compound 4 of the present invention are more rapidly converted to the active metabolic molecule PMPA than the compounds of other structures under the catalysis of human liver microsomes. Resolution of the most active compounds 2 and 4 results in the cis-forms 6-cis and 4-cis, the resolved S-configuration cis compounds and the same type of compound Pradefovir (which has now entered clinical research) Compound 9, S configuration cis) can be metabolized to active molecules faster, and 4-cis is significantly better than 6-cis. The former is metabolized to produce PMPA at a rate 2.2 times that of the latter. Generates a 8.3 times the PMEA rate.

The above results show that the compound 4 in which the benzene ring moiety is substituted by 5-Cl and 2-F, and the compound 2 in which the benzene ring moiety is substituted by 5-F and 3-Cl and the compound 2 are significantly superior to those of other compounds, indicating Compounds with asymmetric substitution of halogen at positions 3, 5 and 2, 5 on the phenyl ring improve the rate at which liver delivery compounds are activated by human liver microsomes.

Example 13 Liver Delivery Compound Experiment

Method

1.1. Animal experiment

Male SD rats weighing 180-300 g were provided by Shanghai Xipuer-Beikai Experimental Animal Co., Ltd. Male animals adapted to the environment for more than 3 days, and fasted for 12 hours in the evening before the experiment. A solution (physiological saline) of PA1010 (4-cis compound), PA1007 (6-cis compound), TAF and TDF was prepared. Before the administration, the animal weight was checked to meet the experimental requirements. Twelve rats were selected for grouping, and 2 rats in each group were given a drug solution of 30 mg/kg by gavage. Rats were euthanized with carbon dioxide gas at 0.5 h, 1 h, 3 h, 6 h, 12 h, and 24 h, respectively. Samples were taken by cardiac extraction, stored in heparin anticoagulant tubes, centrifuged at 6000 rpm for 5 min at 4 ° C, and supernatants were taken. The plasma was stored in ice; the kidney and liver tissues of the rats were collected and washed with 4 ° C pre-cooled saline, and the water was drained. Store in ice. After the experiment, the samples were stored in a -80 ° C refrigerator.

1.2. Determination of PA1010, PA1007, TAF and TDF monophosphate metabolite tenofovir (PMPA) in biological samples

Sample preparation

In the ice bath, the kidney and liver tissues were thoroughly disrupted and mixed with 5 volumes of physiological saline to obtain a tissue homogenate sample. 100 μL of rat plasma or tissue homogenate samples were mixed with 100 μL of an internal 10% trichloroacetic acid precipitant (containing an internal standard of 50 ng/mL adefovir, methanol: acetonitrile = 50:50, v/v). After centrifugation at 6000 rpm for 5 minutes at 4 ° C, all supernatants were treated with SPE micro-extraction plates (MCX μElution Plate 30 μm, Waters), and finally eluted with 5% ammonia in methanol, and 75 μL of the supernatant was taken in a 384-well sample plate. In the injection, 1.0 μL was injected for analysis.

Chromatographic mass spectrometry

LC-MS/MS-AJ (Triple Quad 5500, AB SCIEX) was used for the analysis of the samples. Column: Acquity UPLC HSS T3 (2.1 x 50 mm, 1.8 μm); column temperature: 40 ° C; flow rate: 0.5 mL/min. Mobile phase A: 0.1% aqueous formic acid, mobile phase B: acetonitrile solution. The sample was separated by gradient elution and the procedure is shown in Table 4. And mass spectrometry conditions for the corresponding internal standard: electrospray ionization (ESI) positive ion mode, multiple reaction monitoring (MRM) monitoring ion pair m/z: 288/176 (PMPA), 274/162 (PMEA); capillary voltage It is 3.0 kV; the temperature is 500 ° C; the desolvation gas flow is 1000 L/h; the scanning time is 0.025 seconds; the collision energy is 25V.

Table 4 PMPA liquid phase elution gradient conditions

1.3 Data Analysis

A columnar distribution of the concentration of PMPA in plasma, liver, and kidney corresponding to time. The tissue concentration-time curve area (AUC0-t) of PMPA was fitted using the log-linear trapezoidal method in the non-compartmental model of WinNonLin 6.2.1 (Pharsight, CA). The ratio of liver to kidney to hepatic blood of PMPA is their AUC0-t ratio.

2. Results

After administration of 30 mg/Kg of drug solution by intragastric administration, the distribution of liver tissue showed that the active molecule PMPA released by PA1010 was significantly higher than the TAF and TDF at the corresponding time points (p<0.01, Figures 4, 6 and 7). Using WinNonLin6.2.1 to fit the area under the curve of the drug, according to Table 5, the liver exposure of PMPA released by each test drug was compared: PA1010>TAF>PA1007>TDF, and the PMPA released by PA1010 was 1.5 times of TAF. 222692h·ng/g vs. 148407h·ng/g) and TDF 2.9 times (222692h·ng/g vs. 78050h·ng/g), the results predict that PA1010 can achieve clinical equivalent of TDF and TAF at lower doses. Efficacy. At the same time, the PMPA exposures produced by PA1010 and PA1007 were significantly lower than those of PMPA released by TAF and TDF (Table 5). Combined with the above results, PA1010 and PA1007 showed higher than TAF and TDF at the same dose. Kidney ratio (Figures 4-7 and Table 5), due to the enrichment of PMPA released by TDF and TAF in the kidney The main factor of stage nephrotoxicity, this result predicts that PA1010 and PA1007 at the same dose can significantly reduce the clinical nephrotoxicity caused by TDF and TAF.

Table 5 Exposure of in vivo metabolically released active molecules (PMPA) in plasma, liver and kidney after intragastric administration of 30 mg/kg PA1007, PA1010, TAF and TDF, and the area under the concentration-time curve (AUC0-t) )(h*ng/g)

The above results indicate that the compounds of the formulae I and III of the present invention have higher activity and higher liver-deliverability, which results in lower dosages required for treatment, thus having higher safety or lower toxic side effects. Thus, the clinical therapeutic index of PMPA is greatly improved.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (11)

1. A compound of the formula II below, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof:

   

   among them:

   R _{1 is} selected from hydrogen, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1- a C6 alkylamino group; wherein said substitution has one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, nitro, hydroxy, amino, cyano;

   m is 0, 1, 2, 3, 4, 5;

   Each R _{2 is} independently selected from halogen, nitro, hydroxy, amino, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C6 alkane Oxy, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 carboxyl, substituted or unsubstituted C1-C6 ester, substituted or unsubstituted C2-C6 alkanoyl, substituted or not Substituted C2-C6 alkanoamide group;

   Wherein said substitution has one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, nitro, hydroxy, amino, cyano;

   In Formula II, each chiral center is an R type or an S type.
2. The compound of claim 1 wherein said

   

   From the following group:

   
3. The compound of claim 1 wherein said compound is selected from the group consisting of:

   
4. The compound according to claim 3, wherein said compound has the structure shown below:

   

   

   Wherein R ₃ and R ₅ are halogen;

   n is 0, 1, 2, 3.
5. A compound according to claim 4, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein R ₃ is Cl and R ₅ is F.
6. The compound according to claim 4, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from the group consisting of:

   
7. Salts of the compounds of formula I, formula II and formula III according to claims 1-6 are pharmaceutically acceptable salts of the compounds of formula I, formula II and formula III with inorganic or organic acids, Or a salt of a compound of formula I, formula II and formula III is a pharmaceutically acceptable salt formed by reacting a compound of formula I, formula II and formula III with a base; said formula I, formula II and The compound of the formula III or a salt thereof is an amorphous substance or a crystal.
8. A pharmaceutical composition comprising a therapeutically effective amount of a compound as claimed in any one of claims 1 to 6, or an optical isomer thereof, a pharmaceutically acceptable salt, hydrated Or a solvate; and a pharmaceutically acceptable adjuvant, diluent or carrier.
9. Use of a compound according to claims 1-6, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 8 wherein A pharmaceutical composition for the preparation of an acute or chronic disease associated with the treatment and/or prevention of hepatitis B virus (HBV), hepatitis D virus (HDV) or human immunodeficiency virus (HIV) infection.
10. The use according to claim 9, wherein the acute or chronic disease associated with hepatitis B virus (HBV), hepatitis D virus (HDV) or human immunodeficiency virus (HIV) infection is selected from the group consisting of: Hepatitis, hepatitis D or AIDS.
11. A method of preparing a compound of formula II according to claim 1, comprising the steps of:

    

    i. Condensation of a compound of formula Va and a compound of formula Vc in an inert solvent to provide a compound of formula II.

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