CN101193642A - Nucleotide analogue prodrug and the preparation thereof - Google Patents

Abstract

(R)-9-[2-bis[pivaloyloxymeihoxy]phosphinoylmetlioxypropyl]adenine (being abbreviated bis-POMPMPA, TD), the devivative and the use thereof. Also including the syntheticalprocess of TD and the procedure for manufacturing solid TD, as well as the composition containing TD and the procedure for manufacturing the composition.

Description

Nucleotide analog pro-drug and its preparation

Nucleotide analog pro-drug and its preparation

Technical field

The present invention relates to（R) [2- is double by -9-（Pivaloyloxymethyl）Phosphonylmethoxypropyl] adenine（English name 9- [2- (R)-[bis [pivaloyloxymethoxy]-phosphinoylmethoxy] propyl] adenine, English abbreviation bis-POM PMPA, hereinafter referred to as TD) and its derivative and application.The invention further relates to TD synthetic method and the TD of solid state preparation method, the invention further relates to the preparation method of the composition containing TD and described composition.

Background technology

Phosphonylmethoxy yl nucleosides acid-like substance is broad-spectrum disease resistance cytotoxic compound known to a class, with AntiHIV1 RT activity, HBV, CMV, HSV-1, the activity of the virus such as HSV-2 and human body herpes virus.9- [2- (phosphatidyl methoxy) ethyl] adenosines (abbreviation PMEA) and 9- [(R) -2- (phosphatidyl methoxy) propyl group] adenosine (abbreviation PMPA) are two examples that clinical antiviral therapy is had been used in such compound.Absorption due to phosphonate radical influence human body contained in phosphonylmethoxy yl nucleosides acid-like substance to it, generally requires and changes into lipophilic pro-drug to improve its bioavilability by phosphonylmethoxy yl nucleosides acid-like substance.For example recently by FDA ratify the AD for treating hepatitis B and the tenofovir disoproxil for treating AIDS (tenofovir disoproxil fumarate) be respectively phosphonylmethoxy yl nucleosides acid-like substance PMEA and PMPA lipophilicity prodrug.AD and tenofovir disoproxil can be metabolised to the parent drug PMEA and PMPA for having antivirus action accordingly in vivo.

Tenofovir disoproxil PMPA In recent clinical test, it is found that AD has the adverse reaction of kidney damage.AIDS virus (HIV) can be suppressed when AD was used with the dosage of about 300mg/ days, but found according to relevant drug metabolism dynamics research, AD is ingested after human body and is greatly distributed in kidney organ under 300mg dosage, and renal toxicity is caused to human body;And when AD is respectively with about 50mg/ days, when the dosage of 30mg/ days and lOmg/ days is used, the hepatitis type B virus (HBV) that human body can be suppressed is replicated, but daily 50mg/ days and the adverse reaction of 30 mg groups and the incidence of renal dysfunction are higher, so the treatment that AD can only be with the dosage of the unoptimizable of lOmg/ days for hepatitis B.At present also it has been proposed that, due to the chronicity of Treatment for Hepatitis B, if treatment is longer than 48 weeks, even if using the low dosage of lOmg/ days, if occur and the cumulative toxicity of kidney is also waited to further look at.

The dosage that U.S. FDA ratifies the tenofovir disoproxil treated for AIDS antiviral combination is 300mg/ days, because the dosage that the medicine is used is larger, liver and kidney and other organs to the patient of long-term taking are very big burdens, and make it that the production cost of unit formulation is also higher because the dosage used is larger.

The report of TD grease is only related in existing literature, due to TD grease less stable, and is unfavorable for being prepared into suitable preparation, is used for the ease of it in terms of medicine preparation and storage, often needs to be solidified.So far, also not on solid state TD report also not on by the report of the TD methods solidified.

Summary of the invention

It has now been discovered that compound (R) -9- [double (pivaloyloxymethyl) the phosphonylmethoxy bases of the 2-] propyl group of structure as shown in formula (I)] adenine（TD) there is more excellent antiviral activity and more preferable security compared with AD and tenofovir disoproxil.The compound is the homologue of AD, is also antiviral compound PMPA pro-drug, PMPA can be metabolized in vivo.The English name of the compound is 9- [2- (R)-[bis [pivaloyloxymethoxy]-phosphinoylmethoxy] propyl] adenine, English abbreviation bis-POM PMPA, Chinese name is also referred to as (R) -9- [double (pivaloyl oxygen methyl) the phosphonylmethoxy bases of 2-] propyl group] adenine,（R) -9- [2- double (pivaloyl oxygen methyl) phosphonylmethoxy bases] propyl group] adenine or bis-POM PMPA.

The invention provides：

1) TD and its derivative of solid state, includes TD, TD unformed solidfied material, the TD salt of solid state and the TD cyclodextrin inclusion compound of crystalline state.These TD existed with solid state and its derivative are all easy to industrialize extensive synthesis and with the performance being prepared into required for medicine.

2) TD synthetic method and method of purification, including PMPA is contacted into synthesis TD and post separation method, the method that crystallisation and salt forming method etc. purify TD in polar solvent in the presence of organic base with pivaloyl halo methyl esters.

3) curing of TD grease, including TD grease is changed into TD, TD unformed solidfied material, the TD salt of solid state and TD cyclodextrin inclusion compound of crystalline state.

4) composition and preparation method of the stabilization containing TD and TD derivatives.

5) TD and its derivative of solid state are in the antiviral purposes being particularly in AntiHIV1 RT activity, HBV, CMV, HSV-1, HSV-2 and human body herpes virus.

Detailed description of the invention

TD synthesis and purifying：

PMPA synthesis can be according to existing document, such as Chinese patent application 98807435.4, United States Patent (USP) US5733788 and United States Patent (USP) US6653296 documents.Can also be synthesized according to the method shown in reaction process Fig. 1-

(1) diethyl carbonate is added into reaction vessel,（R)-l, 2- propane diols, plus catalyst sodium alkyl alcohol such as sodium methoxide or caustic alcohol, boil off ethanol, and reaction obtains (R)-carbonic acid -1,2- propylene diester (A);

(2) in the reactor containing inert gas such as nitrogen, add carbonic ester (A) and adenine and N, the alkali such as sodium hydroxide of dinethylformamide (DMF) and catalytic amount, reaction obtains (R) -9- [2- (diethyl phosphono methoxyl group) propyl group] adenine (B);

(3) diethyl phosphite, paraformaldehyde, triethylamine and toluene, heating response 4 ~ 8 hours are added into the reaction vessel for having indifferent gas companion such as nitrogen protection until TLC shows diethyl phosphite disappearance.Reaction solution is cooled to the toluene solution and triethylamine of less than 0 addition paratoluensulfonyl chloride, and reaction completion obtains product p-benzenesulfonyloxymethyl phosphoric acid diethylester (C);

(4) product (B) and DMF that Yi Jia Ru Walk rapid (2) obtains into reaction vessel, 25-75 °C is reduced the temperature to after heating for dissolving, add reaction after lithium hydride and produce (R) -9- (2- hydroxypropyl) Xian Piao Ling lithium salts in 2 hours, p-benzenesulfonyloxymethyl phosphoric acid diethylester (C) is added afterwards, reaction is completed, and obtains (R) -9- [2- (diethyl phosphono methoxyl group) propyl group] adenine (D);

(5) reaction product (D), acetonitrile, front three bromide silicon protective embankment that step (4) obtains are sequentially added into reaction vessel, it is stirred at reflux complete to reaction, volatile liquid is removed in vacuum, residue is dissolved in appropriate water, adjust pH value to 3.0 ~ 3.5, obtain product (R) -9- [2- (phosphonic acids methoxyl group) propyl group] adenine（PMPA).Reaction dissolvent can also select dichloromethane or chloroform, and deprotection agent can also use trimethyl iodine silicon protective embankment or trimethylchloro-silicane to wash/KI.

(A)

Reaction process Fig. 1

TD synthesis and method of purification are as shown in reaction process Fig. 2：

Dry PMPA solids are suspended in polar solvent, organic amine is then added, in order to promote PMPA to dissolve in the reactive mixture, the phase transfer catalyst of catalytic amount can also be added.Reactant mixture stirs addition pivaloyl halo methyl esters after 0.5 2 hours at ambient temperature, reactant mixture is diluted with a large amount of polar organic solvents after being reacted 2 ~ 48 hours under 20 ~ 70 °C, filtering, with weak alkaline aqueous solution and water washing organic phase, dry, the TD crude products that oily is obtained after organic solvent are removed in vacuum.

The above-described preferred DMF and N- methylpyrroles protective embankment ketone (NMP) of polar solvent;The weight ratio scope of PMPA and polar solvent is 1:1~1:20, preferably 1:2~1:10.The preferred trialkylamine of organic amine or Ν, Ν-dicyclohexyl -4- morpholines amidine (DCM), more preferably triethylamine, tri-butylamine and ethyl diisopropyl amine；Organic amine and PMPA mol ratio are 2 ~ 6：1, It is preferred that 3 ~ 4_:The preferred tributyl benzyl ammonium chloride of l o phase transfer catalysts.The preferred pivaloyl chloride methyl esters of pivaloyl halo methyl esters and pivaloyl iodine methyl esters, salt compounded of iodine or bromide are added as the catalyst of substitution reaction to being also an option that property when using pivaloyl chloride methyl esters；Pivaloyl halo methyl esters and PMPA mol ratio are 38_:1, preferably 4 ~ 6： 1.It is preferred that reaction temperature be 45 ~ 65 °C.Dilution organic solvent ethyl acetate or isopropyl acetate, the preferred sodium bicarbonate aqueous solution of weak alkaline aqueous solution.

X=CI, Br, l HA=acid

Reaction process Fig. 2

TD is purified from TD crude products can use following several method：

1) column chromatography：

The use of silica gel is stationary phase, is eluted with 2% ~ 8% methanol dichloromethane solution, collect the component containing TD, decompression boils off the TD purified after solvent.This method purification TD-as be grease, being placed in can slowly decompose at room temperature.

2) crystallisation：

There is a highly polar adenine group to also have two strong lipophilic pivaloyl group in TD molecules, therefore TD can be dissolved in most of polar organic solvents, but the solubility very little in nonpolar or weakly polar organic solvent and water.

TD and solubility can be dissolved it is more than the solvents of 10 mg/ml solvents and be referred to as TD good solvent, it is impossible to dissolves TD or be referred to as TD poor solvent to the solvent that TD solubility is less than 1 mg/ml solvent, the good solvent for the TD that can be selected has：Organic alcohols solvent, organic ketone solvent, esters solvent, halo protective embankment class solvent, organic amide class solvent and organic nitrile solvent and part ether solvent；TD poor solvent has：Alkane solvents, part ether solvent and water.

It is preferred that TD good solvent have：Acetone, butanone, methanol, ethanol, isopropanol, n-butanol, the tert-butyl alcohol, DMF, NMP, acetonitrile, dichloromethane protective embankment, chloroform, ethyl acetate, methyl acetate, isopropyl acetate, Ethyl formate, tetrahydrofuran and oxinane. It is preferred that TD poor solvents have：Methyl tertiary butyl ether(MTBE), di-n-propyl ether, diisopropyl ether, di-n-butyl ether, petroleum ether, just own protective embankment, hexamethylene, pentane, positive protective embankment in heptan and water.

First by TD dissolving crude product in appropriate good solvent, then the solution of gained is mixed with appropriate poor solvent, so that TD solution is close to or up saturation state, then the method such as constitute so that TD solution is supersaturated by changing temperature, evaporation solvent or changing solvent, TD is separated out in the form of crystal.Or TD crude products are directly dissolved in be pre-mixed by good solvent and poor solvent in obtained solvent, form TD solution, crystallization, the TD purified.

TD can be dissolved and the single solvent or mixed solvent that the TD of dissolving separates out with crystalline state can be made to be referred to as TD recrystallisation solvent, the solution formed by TD and its recrystallisation solvent is referred to as TD crystallization solution.Usual TD recrystallisation solvent is one or more kinds of good solvents or the mixed solvent being made up of one or more kinds of good solvents and one or more kinds of non-benign solvents.

It is preferred that TD recrystallisation solvent include above-mentioned all good solvents and optionally from acetone, butanone, methanol, ethanol, isopropanol, n-butanol, the tert-butyl alcohol, DMF, NMP, acetonitrile, dichloromethane protective embankment, chloroform, ethyl acetate, methyl acetate, isopropyl acetate, Ethyl formate, one kind in tetrahydrofuran and oxinane is with optional from methyl tertiary butyl ether(MTBE), di-n-propyl ether, diisopropyl ether, di-n-butyl ether, petroleum ether, just own protective embankment, hexamethylene, positive penta protective embankment, the protective embankment of ring penta, a kind of formed mixed solvent in positive protective embankment in heptan and water, wherein good solvent and not the volume range of good solvent 20:1~1:Between 20.

When good solvent used in recrystallisation solvent is organic alcohols compound and organic ketone compound, preferably ether compound and water are used as non-benign solvent, such as methanol/diisopropyl ether, acetone/diisopropyl ether and ethanol/water mixed solvent.

When good solvent used in crystallization solvent is ester type compound and halogen alkyl compound, preferably alkane derivative is used as non-benign solvent, such as ethyl acetate/just own protective embankment or dichloromethane protective embankment/petroleum ether.

When good solvent used in crystallization solvent is organic amide class compound and organic nitrile compound, preferably water is non-benign solvent.

TD contents in TD crude oil are generally between 5% ~ 60%., can be by TD crude oil in higher temperature is dissolved in the recrystallisation solvent that the good solvent of appropriate amount is constituted when TD comparision contents are high (TD contents are more than 25%), then reduce the crystallization that temperature can obtain TD；And when TD comparision contents are low（25%) TD contents are less than, and generally require with good solvent and the mixture of non-benign solvent formation as recrystallisation solvent.The ratio of usual crystallization solvent and TD crude products is 1:1 and 20:Between 1.

Crystallize the temperature occurred it is general -20 between room temperature, preferably -10 °C ~ 1 (between TC, most preferably 0.C.Relatively low temperature (- 10'C) can improve crystallization yields, but the purity of crystallization is often relatively low；Typically can be while ensure higher yield and purer product being crystallized close under conditions of 0 °C, and the also more convenient and economy in Chemical Manufacture.

3) salt forming method：

It has now been discovered that the salt that TD is formed with acid has good crystal property, the salt pair that usual TD is formed with acid mostly The requirement of crystallization condition crystallizes quantity of solvent used less than relatively low.Therefore, a kind of purification TD method is exactly that TD crude products and appropriate acid first are prepared into salt, then crystallization obtains pure TD salt, pure TD salt is dissolved in appropriate solvent again, neutralized with the weakly alkaline aqueous solution, removing acid group is washed with water, water removal is finally dried, the pure TD that solvent obtains free state is removed.

TD can be into salt with most inorganic acid and organic acid, and the method into salt is：Acid is mixed into salt in appropriate solvent with TD crude products, then separated out salt in crystalline form.The recrystallisation solvent of salt can with it is identical or different into salt solvent, can be again into first removing into salt solvent after salt when different, then the dissolving crude product of TD salt is recrystallized to give to the sterling of TD salt in recrystallisation solvent.

The equivalent of TD in TD crude products is usually slightly larger than into sour equivalent used in salt, the ratio of acid and TD is general 1.1:1~1.3:Between 1.TD amount can be obtained with HPLC methods or UV absorptiometry to determine in TD crude products.

The preferred fumaric acid of salt, maleic acid, salicylic acid or oxalic acid for purifying TD and TD into salt.

In the organic alcohols solvent that TD salt is typically easily soluble in, it can also be dissolved in organic ketone and esters solvent.Neutralized from TD salt obtain the TD of free state-as can use following method：TD salt is dissolved in water-immiscible organic solvent, preferably organic esterses, most preferably ethyl acetate；Then the solution formed is washed into deacidification, the preferred bicarbonate aqueous solution of dilute alkaline aqueous solution with dilute alkaline aqueous solution；After after sour neutralization completely, organic phase uses water or saturated common salt water washing again；Finally dry remove organic solvent just can obtain pure free state TD, now obtain pure free state TD-as in the form of grease exist, for a long time place after can solidify.

The TD and its derivative of solid state preparation and identification：

Due to TD grease less stable, and it is unfavorable for being prepared into suitable preparation, uses, need to be solidified in terms of medicine preparation and storage for the ease of it.The TD of crystallization and unformed solid state, crystal or solid state TD salt and TD cyclodextrin inclusion compound have been prepared now.

(one）The A types crystallization of TD crystallizations, the preparation of unformed solidfied material and identification-I. TD

TD of the present invention A types crystallization refers to the TD crystallizations for being substantially free of water or other solvents, D (X-ray powder diffraction) the spectrum crystal faces of TD A types crystallization represent generally there is peak in 9.774 A, 6.32 A, 5.726 A, 4.967 A, 4.849 A away from d values, it is further typical in 14.917 A, 9.774 A, 6.32 A, the A of 5.726 A. 5.387,5.211 k, 4.967 A, 4.849 A, 4.647 A, 4.553 A, 3.817 A have peak.

(means of differential scanning calorimetry is determined DSC）Endothermic transition temperature is about 100.

IR (infrared absorption spectroscopies）Absworption peak be listed in the table below：

Functional group's absworption peak wavelength

The cm mono- of Ν-Η 3334¹

CH(Ar-H) 3164 cm"¹ Unless otherwise indicated, The TD of anhydrous crystal state accounts for more than the 50% of composition weight, preferably more than 80%, more preferably more than 90%, preferably more than 95% in composition, unformed solidfied material and other crystal formations in addition to the TD of anhydrous crystal state in composition also containing TD.

TD A types crystallization is crystallized under anhydrous conditions by TD to be obtained, and the water content of general recrystallisation solvent used is less than 0.5%.The method of preparation has following several：

1 mixed solvent method：Anhydrous organic ketone or alcohols are used for good solvent, material of organic ethers is that poor solvent constitutes recrystallisation solvent, change the A types crystallization that temperature obtains TD after TD is dissolved.It is preferred that recrystallisation solvent be acetone：Diisopropyl ether volume ratio is 1:2-5 mixed solvent, methanol：Di-n-butyl ether volume ratio is 1:2-10 mixed solvent.The temperature for dissolving TD is 35 60 °C, and crystallization temperature is -20 ~ 35 °C, and preferably -5 ~ 5 °C, crystallization time is 5 4S hours.

2 single solvent methods：Use anhydrous good solvent, it is preferred that acetone, butanone, methanol, ethanol, isopropanol, acetonitrile, dichloromethane, ethyl acetate, methyl acetate, isopropyl acetate, tetrahydrofuran, ether and toluene, pure TD is dissolved by heating, heating-up temperature is usually no more than 50 °C, obtain saturation or the TD solution close to saturation state, then resulting solution is placed under low temperature and crystallizes or resulting solution is placed in room temperature, allow solvent to volatilize naturally, obtain TD A types crystallization.Should be noted it must is when being crystallized using alcohols or ketones solvent, because alcohols or ketones solvent can be with the moisture content in absorption air, it would be possible to obtain the mixture that TD A types crystallization and TD Type B are crystallized, in addition completely TD Type B crystallization.

3 natural coagulation methods：Pure TD is dissolved in anhydrous good solvent, is removed in vacuum and the A types crystallization for obtaining TD is placed after solvent, the A types crystallization for the TD that this method is obtained is mixed with unformed TD sometimes.

II. TD Type B crystallization

TD of the present invention Type B crystallization refers to the TD crystallizations containing two crystallizations water, the XRD spectrum crystal face of TD Type B crystallization is represented generally in 20.157 A, 9.995 A, 4.449 A away from d values, 3.965 A, 3.297 A have peak, it is further typical in 20.157 people, 9.995 A, 5.555 A, 4.696 A, 4.449 A, 3.965 A, 3,677 A, 3.297 A, 3.125 A, 2.822 A have peak.DSC endothermic transitions temperature is at about 55 °C.

IR absworption peak is listed in the table below：

Unless otherwise indicated, TD of the present invention Type B crystallization refers to such composition, two water crystallization state TD account for more than the 50% of composition weight in composition, preferably more than 80%, more preferably more than 90%, preferably more than 95%, unformed solidfied material and other crystal formations in addition to two water crystallization state TD in composition also containing TD.

TD Type B crystallization is separated out by TD in the state of with the presence of water from crystallization solution to be obtained, and at least 0.5% water is contained in general recrystallisation solvent used.The conventional method for preparing TD Type B crystallization is that pure TD is first dissolved in a kind of and miscible good solvent of water, then water is added in the solution formed, so that TD is separated out with the state of crystal, or dissolves pure TD using aqueous good solvent, then crystallize it.

TD A types crystallize in the case where humidity is very high also can moisture absorption be transformed into TD Type B crystallization.

It should be noted that in XRD, the diffraction spectrogram obtained by crystalline compounds is often characteristic for specific crystal formation, wherein bands of a spectrum（Especially in low angle）Relative intensity may because of crystallization condition, the difference of particle diameter and other condition determinations and the advantage orientation effect that produces and change.Therefore, the relative intensity of diffraction maximum is not characteristic to targeted crystal formation, when judging whether identical with known crystal formation, it should be noted that the relative position at peak rather than their relative intensity.Peak position generally is represented away from d with 2 Θ angles or crystal face in XRD spectrum, represents more representative away from d because 2 Θ angles are relevant with the wavelength of incident X-rays, therefore with crystal face.There is simple conversion relation between the two：D=A/2sin9, wherein d represent crystal face away from λ represents the wavelength of incident X-rays（For Cu-K_a, the Α of λ=1.54187), θ is the angle of diffraction.For the crystal formation of the same race of same compound, its XRD spectra has similitude on the whole, and the d values error for characterizing peak position is general within ± 2%, and most of error is no more than ± 1 %;Relative intensity error can be larger, but variation tendency is consistent.In addition, judge crystal formation whether when should be noted that holding organic conception because being not that a diffracted ray represents a thing phase, but a set of specific " " data just represent a certain thing phase.It should also be mentioned that, in the identification of mixture, because the factors such as content decline can cause the missing of part diffracted ray, now, the whole bands of a spectrum observed in high-purity sample need not be relied on, even one bands of a spectrum may also be characteristic to given Tiotropium Bromide crystallization, and A types crystallization crystal face crystallizes crystal face away from the peak for 4.449 A away from the peak for 4.849 people or Type B in such as present invention.

DSC is determined when crystallization is because its crystal structure changes or crystal melt and transition temperature when absorbing or discharge heat.For the crystal formation of the same race of same compound, in continuous analysis, thermal transition temperature and fusing point error are typical within about 5 °C, generally within about 3 °C, when we say that a compound has a given DSC peaks or fusing point, this refers to ± 5 °C of the DSC peaks or Rong points.DSC provides a kind of householder method for distinguishing different crystal forms.Different crystal habits can be recognized according to its different transition temperature feature.It is to be noted that for mixture, its DSC peak or fusing point may change in the larger context.Further, since with decomposition during material melts, therefore fusion temperature and heating rate are closely related.

IR determines infrared absorption caused by the related specified chemical key of the group for corresponding to light in molecule and vibrating.Because the electrical environment of different crystal forms intramolecular covalent bonds is different, covalent bond strength can also be changed, and the change of covalent bond strength is inevitably resulted in The difference of different crystal forms IR spectrum.

III. unformed solidfied materials of TD

The present invention, which is also provided, does not have obvious sharp bands of a spectrum peak in TD unformed solidfied material, the XRD spectrum of the unformed solidification compounds of described TD, only one very wide unformed solid peak.It is also possible to be mixed with a small amount of TD crystallizations in usual TD unformed solidfied material, in general, the content of the unformed solidfied materials of TD is more than 70%.

The preparation methods of the unformed solidfied materials of described TD is following-

1. pure TD is dissolved in good solvent, in substantial amounts of low temperature poor solvent is added in the case of being stirred vigorously, TD is separated out and solidified, and forms the unformed solids of TD.Usually, the temperature of poor solvent is below -20 °C.

2. can also obtain the unformed solidfied materials of TD by solvent is removed with vacuum freeze drying after pure TD dissolvings, the solid powder XRD generally prepared with this method shows the content of the unformed solids of TD more than 70%.

The unformed solids of TD obtained with the method for freeze-drying are usually loose shape solid, and the dissolubility in water is better than the TD of crystalline state, and rate of dissolution is high, are suitable for preparing injection powder injection formulation.

Fig. 7 composes for the x-ray diffractogram of powder of the unformed solids of TD, does not have obvious sharp bands of a spectrum peak in the collection of illustrative plates, only one very wide unformed solid peak.

(-) TD salt

TD and acid reaction generating structure formula following salt or salt form compound： .

Wherein_aIt is the mol ratio of acid and TD, a is between 1 ~ 5, preferably 1 ~ 3, more preferably 1;HA is acid.

Can there should be the acidity for being enough that stable salt is formed with TD with the suitable acid of TD forming salts or salt form compound.Suitable acid can be monoacid or polyacid, including inorganic acid, organic sulfonic acid, organic carboxyl acid and containing acidic-group and with the organic compound or natural products of protection liver effect.

Suitable inorganic acid includes sulfuric acid, phosphoric acid, nitric acid, hydrochloric acid, hydroiodic acid, hydrobromic acid, hydrofluoric acid etc., and suitable organic sulfonic acid includes C₆~₁₆Aryl sulfonic acid, C₆~_I6Heteroaryl sulfonic acid and C_W6 alkyl sulfonic acids, preferably taurine, benzene sulfonic acid, p-methyl benzenesulfonic acid, α-naphthalenesulfonicacid, beta-naphthalenesulfonic-acid,（S)-camphorsulfonic acid, methanesulfonic acid, ethyl sulfonic acid, positive propane sulfonic acid, isopropyl sulfonic acid, positive fourth sulfonic acid, Zhong Ding sulfonic acid, isobutyl sulfonic acid, tertiary fourth sulfonic acid, penta sulfonic acid and own sulfonic acid.Organic carboxyl acid can be unitary or polybasic carboxylic acid, including ~₁₆Alkyl carboxylic acid, C_W6Aryl carboxylic acid and Cw₆Heteroaryl carboxylic acid, preferably acetic acid, glycolic, lactic acid, pyruvic acid, malonic acid, glutaric acid, tartaric acid, citric acid, fumaric acid, butanedioic acid, malic acid, maleic acid, grass Acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, no 1 diacid, cinnamic acid, mandelic acid, salicylic acid and 1- phenoxy benzoic acids, nicotinic acid, pantothenic acid.Organic carboxyl acid also includes amino acid, and suitable amino acid has many, the natural amino acid found especially as protein component, preferably aspartic acid, glutamic acid, valine.

Organic compound or the preferred ascorbic acid of natural products, oleanolic acid, malol, ursolic acid, glycyrrhizic acid, enoxolone, red sage root acid, forulic acid, glucuronic acid, gluconic acid and levulinic acid containing acidic-group and acted on protection liver.Most preferred TD salt has TD fumarates, TD oxalates, TD salicylates, TD oleanolic acid salts and TD aspartates.

The present invention have also obtained the crystallization of TD fumarates, its XRD spectrum crystal face is represented generally in 18.706 people, 6.112A away from d values, 4.562A, 3.645A, 3.56lA, 3.033A, 2.596A has peak, further typical in 18.706 people, 6.112A, 5.075A, 4.562 people, 4.414A, 4.14lA, 4.044A 3.776A, 3.645 A_¾3.561 A, 3.257A, 3.033A, 2.985A, 2.596A have peak.

The absorption of the IR spectrum of the crystallization of TD fumarates about exists： SSllcnT¹, 2979cm"¹ ^ 2941cm^-1, 2879 cm mono-¹、 1752cm^_1、 1683cm— 1304cm"¹ 1142cm"¹, 980cm— ¹There is peak.

It is typically to mix TD in the solution according to the ratio into salt with acid to prepare TD salt, and used acid can also be slightly excessive.Solvent typically selects organic alcohols, when acid is inorganic acid or organic sulfonic acid and some water miscible sour such as amino acid, solvent can from ~₄Alcohol, water or water and the mixed solvent of organic solvent formation.To some fat-soluble strong acid such as oleanolic acids, malol etc. can make solvent into during salt with alkyl halide and esters.After TD is mixed in a liquid with acid, in the case of stirring or cooling, the crystal of salt can be separated out.Solvent evaporation in the solution of TD salt can generally also be obtained into TD salt solids, these solids can be the mixture of the unformed solid of crystal or TD salt or both.

TD salt exists with solid state mostly.Many TD salt has fusing point high compared with TD, and stability is good, and the characteristics of easily forming crystalline solid is conducive to industrialized production and storage, is also beneficial to the preparation and storage of preparation.TD salt or salt form compound still has and TD identical antiviral activities; and if TD be combined with containing acidic-group and with protection liver effect organic compound or the salt or salt form of natural products formation; then these salt can retain original antiviral activity, but also with liver protection function.Therefore, TD salt or salt form are compound may also be used for preparing antiviral drugs.(three）TD cyclodextrin inclusion compound

Cyclodextrin be by 6,7 or 8 glucose molecules are with 1, the cyclic oligomer carbohydrate compound of 4- glycosidic bonds connection, is water miscible irreducibility white crystalline powder, and structure is hollow round flow-through, it is in hydrophily at the opening in hole, the inside in hole is in very strong hydrophobicity.Many molecules can be embedded in interior formation supramolecular structure by cyclodextrin molecular bag.

Medicine, which is made after inclusion compound, using cyclodextrin to solidify liquid drug, improve the stability of medicine, increase the solubility of medicine, improve the bioavilability of medicine.

We have found that TD can form inclusion compound with cyclodextrin, and because lipophilic pivaloyl group is embedded into cyclodextrin After in hydrophobic cavity; not only so that pivaloyl group is more difficult to hydrolysis to improve TD stability; and improve solubility and dissolution velocity of the TD in water, it is possible to increase and the dissolution rate and bioavilability of preparation, also allow for being prepared into the solution type preparations such as injection.

Described TD cyclodextrin inclusion compounds are TD and cyclodextrin in molar ratio 1:1~1 :10 inclusion compounds formed, preferably 1:1-1:3；Described cyclodextrin is α cyclodextrin or derivatives thereof, beta cyclodextrin or derivatives thereof, γ cyclodextrin or derivatives thereof, preferably beta cyclodextrin or derivatives thereof, most preferably beta cyclodextrin.

TD cyclodextrin inclusion compound can be in the liquid phase obtained by mixing by by TD with cyclodextrin, and adoptable preparation method includes saturated water solution method, polishing, freeze-drying and ultrasonic method etc..

1) saturated water solution method

The TD organic solvents such as appropriate alcohols or ketone are dissolved, by weighing cyclodextrin with 1 ~ 10 times of TD mol ratios amount and be made into 50-80 saturated aqueous solution, two kinds of solution are mixed into more than 30min, freezing makes inclusion compound Precipitation, filtering, is cleaned with organic solvents such as appropriate alcohols or ketone, is drying to obtain.Alcohols or the preferred methanol of organic solvent of ketone, ethanol, isopropanol and acetone.

2) polishing

By a certain amount of TD, the cyclodextrin of 1 ~ 10 times of amount of addition after being dissolved with organic solvents such as appropriate alcohols or ketones adds suitable quantity of water mixing, is fully ground into pastel, after low temperature drying, then is produced with the organic solvent cleaning, drying such as alcohols or ketone.

3) freeze-drying

By TD and cyclodextrin in molar ratio 1:1 ~ 10 weighs, and is dissolved in the water containing the organic solvent such as 0 ~ 20% (v/v) alcohols or ketone, stirring and dissolving is simultaneously degerming by miillpore filter, puts to refrigerate in liquid nitrogen container and be freeze-dried about 24h again, produces.

TD Benexate Hydrochloride is dissolved in after water, with after 6% methanol dichloromethane solution expansion on TLC, TD Benexate Hydrochloride is found in origin in ultraviolet lower fluorescence developing, Rf values are 0, and free TD Rf values are 0.4.Above-mentioned qualification result all illustrates that TD and beta-schardinger dextrin form inclusion compound.

The dissolubility and stability of TD and its derivative after solidification contrast following-dissolubility analysis

Tested with reference to two notes on the use of China's coastal port, precision weighs sample lg, is slowly added to a certain amount of solvent, every strength shaking in 5 minutes 30 seconds, observe the dissolving situation in 30 minutes, as a result see the table below-solubility experiment

Solvent methanol absolute ethyl alcohol 0.1N HC1 water O.lN NaOH quantity of solvent TD A types crystallization 1.4 5.0 33.5 880 910

(ml) TD Type B crystallization 1.5 6.0 27.3 840 790

Unformed the 2.0 5.6 25.2 380 613 of TD TD fumarate 7.4 15.5 18.5 120 65

TD salicylate 8.2 12.0 23.6 75 82.5

TD oxalates 84.3 128.3 19.8 89.5 91.2

TD oleanolic acid salt 760 650>1000 >1000 >1000

TD beta cyclodextrin clathrate> 1000 > 1000 6.5 7.4 7.0

The TD readily soluble dissolving slightly soluble slightly soluble of Α types crystallization

The TD readily soluble dissolving slightly soluble slightly soluble of Type B crystallization

TD unformed readily soluble dissolving dissolving slightly soluble

The readily soluble dissolving dissolving dissolving conclusion TD of the TD fumarate readily soluble dissolving dissolving dissolving of salicylate

TD oxalates dissolving dissolving dissolving dissolving dissolving

TD oleanolic acid salt slightly soluble slightly soluble is insoluble insoluble

The insoluble readily soluble readily soluble analysis of stability η of TD beta cyclodextrin clathrate-

(-) exposure experiments to light

Sample is uniformly shared into open culture dish, thickness≤5mm, adjustable range, it is 4500 ± 500Lx to make intensity of illumination, respectively at 5, sampling detection in 10 days, and compareed, as a result see the table below with the result of 0 day：

Exposure experiments to light (4500 ± 500Lx)

Note：23-26 °C of temperature change;Relative humidity variations 56%-63%.

(：Two) hot test

Sample is positioned in sealing clean vial, is placed in 60 °C of thermostatic drying chambers, respectively at sampling detection in 5,10 days, and is compareed with the result of 0 day.As a result see the table below- Hot test（60 °C) 54%-62 ° of relative humidity variations/₀

(3) high wet test

Sample is hooked and shared into open culture dish, thickness≤5mm is placed in room temperature（25 °C), relative humidity is in 75 ± 5% constant temperature and humidity incubator, was measured, and compareed with the result of 0 day respectively at sampling in 5,10 days.As a result it see the table below：

High wet test（Room temperature, 23-26 °C of temperature change of relative humidity 75 ± 5%)

Time（My god）

Investigation project

0 5 10

TD A types crystallization 4.3 5.6

TD Type B crystallization 0 0.2

TD unformed 1.0 2.3 moisture absorption weightening TD fumarate 00

(%) TD salicylate 00

TD oxalates 00

TD oleanolic acid salt 00

TD beta cyclodextrin clathrate 0.2 0.7

TD Α types crystallization 99.7 98.5 97.2

TD Type B crystallization 99.6 98.2 96.9

TD unformed 99.0 97.9 95.8 content TD fumarate 99.2 99.1 99.1

(%) TD salicylate 99.1 99.1 99.0

TD oxalates 99.4 99.3 99.2

TD oleanolic acid salt 99.2 99.1 99.1

TD beta cyclodextrin clathrate 99.4 99.2 99.2 TD A types crystallization 96.3-97.1 89.2-90.7 86.1-90.7

The TD 2-64.7 of Type B crystallization 63.5-64.5 62.8-65.2 62.

TD's is unformed

TD fumarate 118.7-119.1 118.0-119.0 118.3-118.8 fusing point rc)

TD salicylate 87.3-88.3 87.5-87.9 87.9-88.3

TD oxalates 153.5-154.0 152.8-153.7 153.4-153.9

TD oleanolic acid salt 242.3 (decomposition) 242.0 (is decomposed）241.6 (decompose）

TD beta cyclodextrin clathrate 312.3 (decomposition) 312.0 (decomposition) 312.1 (decomposition)

(：Four:) accelerated test

Sample is packed with polyethylene film plastic bag sealing, 40 ± 2 °C are placed in, relative humidity is in 75 ± 5% constant temperature and humidity incubator, places 3 months, respectively at 1,2,3 the end of month sampling detections, and is compareed with the result of 0 month.As a result it see the table below：

Accelerated test（4 (TC, relative humidity 75%)

From the above results, all TD and its derivative that the present invention is obtained have good stability, are adapted to be prepared into any form of composition or pharmaceutical preparation, especially the TD crystallization of A types and TD salt.With TD crystal and solid phase ratio, most of TD salt and TD cyclodextrin bag and thing have good water solubility, can be prepared into pharmaceutical solutions, including primary infusion, liquid drugs injection, oral liquid or powder pin.

Method of administration and Pharmaceutical composition

The TD or its physiologically acceptable derivative that the present invention is provided include:TD A types crystallization, TD Type B crystallization, the unformed solidfied materials of TD, TD salt form compound and cyclodextrin inclusion compound, can be by the way of any suitable treated disease Footpath is administered.Usually, TD or its physiologically acceptable derivative can pass through per rectum, vagina, intranasal, part（Including eyes, oral cavity and sublingual）And parenteral（Including subcutaneous, muscle, intravenous, intracutaneous, intrathecal and exterior dura）Etc. approach administration, preferably it is administered orally.

Although TD or its physiologically acceptable derivative can be administered in the form of pure material, it is administered generally in the form of TD pharmaceutical preparation.TD pharmaceutical preparation includes TD or its physiologically acceptable derivative and one or more pharmaceutical carriers; optionally; other treatment composition or auxiliary element can also be contained, for example other antivirotics, immunopotentiating agent, protection liver drug and L-BETAIN and its salt etc..Pharmaceutical carrier includes adhesive, diluent, disintegrant, preservative, dispersant, glidant（Antitack agent）And lubricant.

The solid pharmaceutical preparation of suitable oral TD or its physiologically acceptable derivative includes tablet, capsule, pulvis, granule, dripping pill, powder, bolus, tincture or paste etc.；Wherein tablet is conventional tablet, dispersible tablet, effervescent tablet, sustained release tablets, controlled release tablet or enteric coatel tablets, and capsule is conventional capsule, spansule, controlled release capsule or capsulae enterosolubilis.

The unit formulation of the tablet and capsule of TD or its physiologically acceptable derivative contains TD5 ~ 300mg, preferably 5 ~ 150mg.In addition to active ingredient, generally also contain appropriate filler, such as starch, sucrose and lactose；Adhesive, such as water, ethanol, polyvinyl pyrrolidone and pre-gelatinized starch；Disintegrant, such as microcrystalline cellulose, Ac-Di-Sol, cross-linked pvp；Lubricant, such as magnesium stearate, talcum powder or silica pharmaceutical carrier.It can also contain formaldehyde scavenger in addition（Such as lysine or gelatin）So that catch can releasable formaldehyde in TD storage processes.

Alkaline pharmaceutical carrier, including basic carbonate and alkaline hydrated oxide can also be contained in the tablet and capsule of TD or its physiologically acceptable derivative.It is preferred that basic carbonate be calcium carbonate, magnesium carbonate, zinc carbonate, ferrous carbonate and aluminium carbonate；It is preferred that alkaline hydrated oxide be magnesium hydroxide, calcium hydroxide, aluminium hydroxide and iron hydroxide.These alkaline pharmaceutical carriers can improve the stability of TD in preparation, reduce TD degraded.

The preparation of TD or its physiologically acceptable derivative can also contain VBT or its salt（Such as VBT tartrate (2:1 )).TD is metabolized the pivalic acid of generation and seems to make the VBT concentration in patient's body to decline in vivo.Preparation containing VBT or its salt and TD can reduce effect of the pivalic acid in terms of the VBT in TD patient's body is taken in reduction.The amount of the L- carnitines of addition can be determined according to the depletion degree of VBT in patient's body.

' the dispersible tablet of TD or its physiologically acceptable derivative can reach quickly disintegrated purpose containing about 0.5 ~ 60% disintegrant；Contain enteric material in TD enteric coatel tablets or be coated with enteric-soluble coating material; capsulae enterosolubilis preparation can be the particle or piller that are coated by enteric material that the capsule made of enteric material is packed the capsule preparations made or packed with conventional capsule.

The tablet and capsule of TD or its physiologically acceptable derivative can be prepared with method general in pharmacy.Tabletting after water or ethanol wet granulation can be used by preparing tablet, can also use dry powder direct tabletting, and capsule can be filling after first wet granulation, can also be directly filling with dry powder. TD or its physiologically acceptable derivative can also be administered in the way of injection, and preparation includes injection sterile powder and injecting fluid.

TD bioactivity

First, acute toxicity test, using median lethal dose (LD_5Q) experiment

TD fumarates and TD A types crystallization are dissolved in 0.1M aqueous citric acid solution respectively, and animal uses body weight

18 ~ 22g health Kunming mouses 140, randomized blocks is divided into 14 groups of every group 10, male and female half and half.

Intragastrically, by trial test result, TD fumarates and TD A types crystallization calculate LD respectively with 7 administration group various dose gastric infusions, Continuous Observation 14 days, the reaction of observation mouse toxicity and death condition₅₍₎。

The LD of TD fumarates_5QFor 6.05g/kg, 95% credible is limited to 4.50 7.87g/kg.

The LD of TD A types crystallization₅Q is 4.31g/kg, and 95% credible is limited to 2.83 5.44g/k.

2nd, long term toxicity test

Using BEAGLE dogs as animal model, using AD as reference substance, the long term toxicity of TD A types crystallization is investigated, high spot reviews TD A types crystallize the influence to renal function.

30 BEAGLE dogs are randomly divided into 5 groups, every group 6, one of which is blank control group, three groups be respectively TD A types crystallization basic, normal, high three dosage groups, the dosage of low dose group be 5mg/kg 1 time a day, the dosage of middle dose group for 15mg/kg 1 time a day, the dosage of high dose group for 45mg/kg 1 time a day, another set be AD control group, dosage be 40mg/kg 1 time a day.

The dosage that medicine should be taken by each dog is with drug after salad oil mixing, and successive administration thing 6 months stops observation post administration 21 days.

Each group animal medication therebetween and convalescence no abnormality seen performance, animal unexpected death does not occur, it is low that hematology and hematuria biochemics inspection Check have found that blank control group and TD A types are crystallized, in, there are no significant for every hematological indices and hematuria biochemics standard difference between high three dosage groups, but the serum creatinine of AD and the index of urea nitrogen are significantly raised, illustrate that long-term taking AD has Nephrotoxicity, and the TD of same dosage A types crystallization is then safe, it see the table below：

3rd, Antiviral assay in vivo

Internal anti-hepatitis B virus experiment, observation are carried out using the sheldrake of the vertical infection duck hepatitis B virus at two monthly ages Drug effect.80 Gaoyou sheldrakes are randomly divided into 8 groups every group 10, wherein three groups are administered TD fumarates 5,15,45mg/kg 1 time a day respectively, three groups are administered tenofovir disoproxil respectively in addition（Tenofovir disoproxil fomarate) 5,15,45mg/kg 1 time a day, remaining one group of administration AD 15mg/kg 1 time a day, last group be blank control group.Every 7 days 28 days sampling blood of administration determines the inhibition to DHBV- DNA levels with PCR methods, and inhibiting rate see the table below.Experimental result shows that TD interior resisting virus activity is far above tenofovir disoproxil and AD.

4th, pharmacokinetics and tenofovir is distributed in vivo

1st, bioavilability

Mouse 10, is randomly divided into two groups, every group 5, difference gastric infusion 1H-TD fumarate 30mg/lcg, radiological dose is 135 Ci/kg_;Tenofovir disoproxil 30mg/kg, 135 Ci/kg, take determination of plasma radioactivity to be converted into blood concentration in different time.

The concentration of-TD fumarates and tenofovir salt in blood plasma（Ug/ml)-time compares.

2nd, distribution in the tissue

From Wistar rats 30,6 groups are randomly divided into, after fasting 12 hours, 3 groups pass through gastric infusion 20mg/kg's_TDFumarate, 3 groups of tenofovir disoproxil fumarates by gastric infusion 20mg/kg in addition, in administration 1,4,8 hours live in killing through femoral artery bloodletting respectively and take TD fumarates and tenofovir disoproxil fumarate（Control group）Each one group of animal.The liver and kidney of animal are taken respectively, tissue assay balance claims weight in wet base, 1 is pressed with distilled water:3 prepare homogenate, and 1000g is centrifuged 10 minutes, takes supernatant.Animal organ is hooked into slurry Supernatant samples 0.25ml to be placed in tool plug teat glass, plus Enter the PMEA aqueous solution of 50 microlitres of redistilled water and concentration for 10mg/L（Inner mark solution）50 microlitres.0.5 milliliter of methanol is added after well mixed, vortex is centrifuged 10 minutes after 1 minute（3000r/min), 20 microlitres of concentration with the PMPA in HPLC/MS-MS tissue of supernatant is taken.

The chromatographic condition of Liquid Chromatography/Mass Spectrometry is as follows：

Chromatographic column is Diamonsil C-18 posts, 250mmX4.6mm, 5 micron grain sizes；Mobile phase is methanol-water-formic acid（20: 80： 1 )；Flow velocity 0.5mL/min.

Mass Spectrometry Conditions：

U.S. Finnigan TSQ types chromatography-mass spectroscopy-GC-MS.Ion gun is ESr sources, source voltage 4.5kV;Collision induced dissociation voltage is 40eV;Positive ion mode is detected；Ionic reaction for quantitative analysis is m/z288 → m/zl76.From PMEA as internal standard, ionic reaction is m/z274 → m/zl62.

The distribution of TD fumarates and tenofovir disoproxil fumarate in the tissue is compared

Explanation：

TD groups refer to the animal groups for taking TD fumarates, and control group refers to the animal groups for taking tenofovir disoproxil fumarate.

Rat takes after same amount of TD fumarates and tenofovir disoproxil fumarate respectively, the concentration for the PMPA that the concentration ratio the latter of the PMPA that the former produces in liver produces is higher by about 70% ~ 100% respectively at different time points, and from the point of view of the ratio that liver kidney is distributed, take after TD fumarates, concentration of the PMPA in liver is about 4 times of the PMPA concentration in kidney, and takes 2.5 times that concentration of the PMPA in liver after tenofovir disoproxil fumarate is about the PMPA concentration in kidney.Therefore, its metabolite PMPA that TD fumarates can make significantly is enriched with liver, with hepatic targeting.

Brief description of the drawings

Fig. 1:TD nuclear magnetic resonance（1H-NMR) collection of illustrative plates

Fig. 2:TD mass spectrum（MS) collection of illustrative plates

Fig. 3:The XRD spectrum of TD A types crystallization

Fig. 4_:The DSC collection of illustrative plates of TD A types crystallization

Fig. 5:The IR collection of illustrative plates of TD A types crystallization

Fig. 6:The XRD spectrum of TD Type B crystallization

Fig. 7:Thermogravimetic analysis (TGA) (TG) collection of illustrative plates of TD Type B crystallization

Fig. 8:The DSC collection of illustrative plates of TD Type B crystallization Fig. 9:The IR collection of illustrative plates of TD Type B crystal

Figure 10：The XRD spectrum of the unformed solidfied materials of TD

Figure 11:The ^-NMR collection of illustrative plates of TD fumarates

Figure 12:The IR collection of illustrative plates of TD fumarates

Figure 13:The XRD spectrum of TD fumarates

Figure 14:The 1H-NMR collection of illustrative plates of TD oxalates

Figure 15:The IR collection of illustrative plates of TD oxalates

Figure 16:The XRD spectrum of TD oxalates

Figure 17_:The IR collection of illustrative plates of TD salicylates

Figure 18:The IR collection of illustrative plates of TD oleanolic acid salts

Embodiment：

The synthesis of embodiment 1 (R)-carbonic acid -1,2- propylene diesters：

To diethyl carbonate（380ml, 15.1 moles）(9g sodium methoxides are dissolved in the solution in 50ml absolute ethyl alcohols with denatured ethyl alcohol 40ml is added in the mixture of 200g (R) -1,2- propane diols）, solution then is heated into 8, and (TC slowly boils off ethanol.The process of reaction detects with TLC, until TLC show it is micro or when can not detect (R)-l, 2- propane diols untill.Extremely ooze, then distilled with vavuum pump without ethanol in 120 water pump vacuum distillations, obtain 111 grams of colourless transparent liquid, yield 81.2%, product purity is 97% through GC analyses.

The synthesis of the p-benzenesulfonyloxymethyl phosphoric acid diethylester of embodiment 2：

In inert gas（Nitrogen）Protection under, by toluene（200ml), diethyl phosphite（400ml), paraformaldehyde (120g) and triethylamine（50ml) it is mixed and heated to 70 °C and reacts 2 hours, is then warming up to after backflow and continues to react, until with TLC (solvent normal hexanes：Ethyl acetate=1:4) show micro or can not detect that reaction terminates during diethyl phosphite.Solution is cooled to less than 10 °C, adds paratoluensulfonyl chloride（560g), then it is slowly added into triethylamine under about 5（560ml), temperature is maintained to be no more than 10 °C.Be warmed to room temperature after completion of dropping, react 8h, until TLC show it is micro or when can not detect to toluene semi-annular jade pendant acyl chlorides untill.Suction filtration removes solid, and solid is washed with q. s. toluene.Cleaning solution and filtrate use 5%NaC0 after merging₃The aqueous solution and water wash paint 2 times respectively, and solvent is boiled off at a temperature of not higher than 50 °C after anhydrous sodium sulfate water removal, obtain 600g colourless liquids, and purity is 86% through GC analyses, purified can not be directly used in subsequent reactions.

The synthesis of embodiment 3 (R) -9- [2- (diethyl phosphono methoxyl group) propyl group] adenine

In inert gas（Nitrogen）Protection under add adenine（100g), sodium hydroxide（1.2g),（R)-carbonic acid -1,2- propylene diesters（84g), and Ν, Ν-dimethylformamide（700ml), reactant mixture is stirred 30 hours until TLC (10%MeOH CH at 130 °C₂C1₂Solution（Volume ratio））Untill the remaining adenine of display is not higher than 0.5%.Reactant mixture is cold But to 25 °C, lithium hydride is added（8g), 70 °C are heated under nitrogen protection to react 2 hours, room temperature is subsequently cooled to, and add p-benzenesulfonyloxymethyl phosphoric acid diethylester（300_g), reactant mixture maintains 60 °C untill TLC shows reaction completely.Reactant mixture is concentrated in vacuo at a temperature of no more than 80 °C, is added water（500ml) dissolve, the aqueous solution is continuously extracted with dichloromethane protective embankment, merge dichloromethane protective embankment extract, not higher than 80^DExtract is concentrated in vacuo under C, viscous orange oil 200 is obtained_g, HPLC analyses, which are shown in orange, contains 65% (R 9- [2- (diethyl phosphono methoxyl groups）Propyl group] adenine, being somebody's turn to do (R) -9- [2- (diethyl phosphono methoxyl group) propyl group] adenine crude product purified can not be directly used in subsequent reactions.

Embodiment 4 (R) -9- [2- (phosphonic acids methoxyl group) propyl group] fast cry of certain animals of gland（PMPA synthesis)-

(R) -9- [2- (diethyl phosphono methoxyl group) propyl group] adenine crude product (100g) is dissolved in acetonitrile (122ml), bromination trimethyl silicane protective embankment (207g) is added under nitrogen protection, reactant mixture flows back 4 hours under 70 °C, TLC shows that raw material is depressurized after being wholly absent and boils off solvent, residue 200ml water dissolves, it is cooled to 20 °C, Lip river is washed with dichloromethane or ethyl acetate, aqueous phase adjusts PH to 3.1 ~ 3.5 with 50% sodium hydrate aqueous solution, room temperature is slowly stirred about 3 hours, solid is collected by filtration, cold water is used respectively（50ml) and acetone（50ml) wash, obtain PMPA solid crude products 60g.90 ° of 0 pure water 200 is added into PMPA solid crude products!1, be cooled to room temperature after being sufficiently stirred for, filter, continuously washed with cold water and acetone after standing overnight, be dried in vacuo under 50 °C, obtain 45 grams of PMPA, HPLC analyses show purity be 99 °/..

Embodiment 5 (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine（TD synthesis) and silicagel column separating-purifying：

Under nitrogen protection by solid PMPA (40g) and anhydrous N-N dimethylformamides（160ml) and triethylamine（120ml) mix, be slowly stirred the suspension of gained and add chloromethyl pivalate after being heated to 50 °C, 1 hour（60ml), temperature maintains 50 ~ 55 °C, reacts and cools down for 8 hours or so, adds ethyl acetate（4000ml), it is sufficiently stirred for, is filtered to remove solid, filtrate uses 5%NaHC0₃Washed respectively with water 2 times, the dry water removal of anhydrous sodium sulfate thousand, at a temperature of not higher than 50 °C, organic solvent is removed in vacuum, viscous yellow oil 47g is obtained, containing about (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine 55%.Take 200 grams of 200-300 mesh silica gel, dry column-packing, by grease（Dry method after proper silica gel 47g) is wrapped up in be loaded.Eluted successively using the mixed liquor of 5%-10% ethanol/methylene, collect the eluent containing TD, after component merges, boil off solvent, TD the grease 18.0g, HPLC purified shows that purity is 95.2%.

'H-NMRCCDCb): 8.347(lH,s,H-8)、 7.969(lH,s,H-2)、 5.819(2H,s,N¾)、 5.676(4H,m,CH₂), 4.360 OP (lH, dd, J=14.4,2.8, H-l), 4.132 (lH, dd, J=14.4,7.2, H-r), 3.933 (lH, m, H-2),

3.898 (lH, dd, J=14.0,8.8, H-4), 3.677 (lH, dd, J=14.0,9.2, H-4'), 1.238 (3H, D, J=6.0, C), 1.215 (18H, d, J=6.0, CH₃) (Fig. 1)

MS:Molecular ion peak m/e: 516.2 (M+H⁺) , 538.2 (M+Na⁺) (Fig. 2)

UV-VIS (methanol）：Maximum absorption band 260nm. Embodiment 6 (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] fast cry of certain animals of gland（TD synthesis) and crystallization and purification are by PMPA (40g) and 1-METHYLPYRROLIDONE（160ml), ethyl diisopropyl amine（Iodometyl pivalate is added after 140ml) being mixed and heated to 50 °C, 30 minutes under nitrogen protection（65ml), temperature maintains 50 ~ 55 °C, cold after reacting 4 hours to lack to room temperature.In the reactive mixture plus 4000ml ethyl acetate, it is sufficiently stirred for, is filtered to remove solid, filtrate uses NaHC0₃And water（Each 200ml) respectively to wash three times, anhydrous sodium sulfate drying water removal at a temperature of not higher than 50 °C, is evaporated in vacuo organic solvent, obtains viscous yellow oil 66g.HPLC shows that the content of (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine is about 38%.Use methanol（Grease 200ml) is dissolved, water is added（White solid is obtained after 800ml), is washed after filtering with the ethanol freezed on a small quantity, room temperature in vacuo obtains TD solids 21g, HPLC and shows that purity is 96.3% after drying.

Embodiment 7 (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] fast cry of certain animals of gland（TD synthesis and crystallisation) is purified PMPA (40g) and 1-METHYLPYRROLIDONE（160ml), triethylamine（120ml) with (1 gram of tributyl Benzylphosphonium Bromide ammonium）Addition pivalic acid methyl esters after 50 °C, 30 minutes is mixed and heated under nitrogen protection（60ml), temperature maintains 50 ~ 55 °C, and reaction 8 hours or so is rear cold to be lacked to room temperature.In the reactive mixture plus 4000ml ethyl acetate, it is sufficiently stirred for, is filtered to remove solid, filtrate uses NaHC0₃And water（Each 200ml) respectively to wash three times, anhydrous sodium sulfate drying water removal at a temperature of not higher than 50 °C, is evaporated in vacuo organic solvent, obtains viscous yellow oil 53g.HPLC shows that the content of (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] fast cry of certain animals of gland is about 56%.Use acetone（Yellow oil 200ml) is dissolved, isopropyl ether is added（800ml), room temperature is cooled to after mixing, crystal seed is added, it is positioned over and (white crystals is obtained after TC24 hours, are washed after filtering with a small amount of isopropyl ether, solid 26g is obtained, XRD analysis are shown as TD A types crystallization, and HPLC shows that purity is 98.9%.

Embodiment 8 (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine（TD synthesis and crystallisation) is purified PMPA (40g) and 1-METHYLPYRROLIDONE（160ml), triethylamine（Chloromethyl pivalate is added after 120ml) being mixed and heated to 50 °C, 30 minutes under nitrogen protection（60ml), temperature maintains 50 ~ 55 °C, and reaction 12 hours or so is rear cold to be lacked to room temperature.In the reactive mixture plus 4000ml ethyl acetate, it is sufficiently stirred for, is filtered to remove solid, filtrate uses NaHC0₃And water（Each 200ml) respectively to wash three times, anhydrous sodium sulfate drying water removal at a temperature of not higher than 50 °C, is evaporated in vacuo organic solvent, obtains viscous yellow oil 49g.HPLC shows that the content of (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine is about 52%.Use acetone（Yellow oil 200ml) is dissolved, n-butyl ether is added（800ml) it is positioned over after 0 24 hours and obtains white crystals, washed after filtering with a small amount of n-butyl ether, obtain solid 22g, XRD analysis is shown as TD A types crystallization, and HPLC shows that purity is 98.3%.

Embodiment 9 (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine（TD synthesis) and salt forming method purification wash PMPA (40g) and N- methylpyrroles ketone（160ml), triethylamine（Chloromethyl pivalate is added after 120ml) being mixed and heated to 50 °C, 30 minutes（60ml), 50 ~ 55 °C of temperature maintenance, reaction 8 hours or so, in the reactive mixture Power mouthful 4000ml ethyl acetate, is sufficiently stirred for, is filtered to remove solid, filtrate uses NaHC0₃And water（Each 200ml).After water removal at a temperature of not higher than 50, organic phase is evaporated in vacuo, viscous yellow oil 48g is obtained.HPLC shows TD contents about 56% in grease.Grease methanol（After 100ml) dissolving, fumaric acid solution is added（7g is dissolved in 100 ml methanols）, it is placed in that (TC is stirred overnight, and is filtrated to get 29 grams of TD fumarates.Gained fumarate is dissolved in ethyl acetate, with the Na H C O of saturation₃Aqueous solution 200ml is washed 3 times, is washed with water to neutrality, is separated aqueous phase discarded.Organic phase is removed water, at a temperature of not higher than 50 °C, and vacuum distillation organic phase obtains TD grease 21g, and grease is progressively solidified into solid-like TD after room temperature is placed.Solid powder is ground to obtain after solid vacuum drying, XRD analysis show that solid crystallizes for TD A types, and HPLC shows that TD purity is 99.1%.

Embodiment 10 (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine（TD synthesis and salt forming method) is purified PMPA (40g) and 1-METHYLPYRROLIDONE（160ml), triethylamine（50'C 120ml) is mixed and heated to, chloromethyl pivalate is added after 30 minutes（60ml), temperature maintains 50 ~ 55 °C, reacts 8 hours or so, the π ethyl acetate of power 4000, is sufficiently stirred in the reactive mixture, is filtered to remove solid, and filtrate uses NaHC0₃And water（Each 200ml).After water removal at a temperature of not higher than 50, organic phase is evaporated in vacuo, viscous yellow oil 60g is obtained.HPLC shows TD contents about 38% in grease.Grease acetone（After 100ml) dissolving, oxalic acid solution is added（5g is dissolved in 100 ml methanols）, it is placed in 0 °C overnight, is filtrated to get 24 grams of TD oxalates.Gained oxalates is dissolved in ethyl acetate, with the NaH C O of saturation₃Aqueous solution 200ml is washed 3 times, is washed with water to neutrality, is separated aqueous phase discarded.Organic phase is removed water, at a temperature of not higher than 50 °C, and vacuum distillation organic phase obtains TD grease 19g, and grease is progressively solidified into solid-like TD after room temperature is placed.XRD analysis show that solid is that (A types are crystallized TD）With unformed TD mixture, HPLC shows that TD purity is 99.3%.

The preparation of the TD of embodiment 11 A types crystallization

95% 2 grams of TD grease is dissolved in absolute methanol at about 35 °C（In 10ml), isopropyl ether is instilled to this solution under stirring condition（In 30ml), -4 °C are placed in until separating out solid, filtering, vacuum drying obtains the A types crystallization that TD crystallization l.'38g, XRD analyses are shown as TD, and HPLC analyses show that purity is 98.5%.

The preparation of the TD of embodiment 12 A types crystallization

95% 2 grams of TD grease is dissolved in anhydrous THF (6ml) at about 40 °C, place at room temperature until separating out solid, filtering, vacuum drying obtains TD crystallizations 1.62g, XRD analysis are shown as TD A types crystallization, and HPLC analyses show that purity is 97.8%.

The preparation of the TD of embodiment 13 A types crystallization

95% 0.5 gram of TD grease is dissolved in dry toluene at about 60 °C（In 60ml), place at room temperature until separating out solid, filtering, vacuum drying obtains TD crystallization 0.42g, XRD analysis are shown as TD A types crystallization, and HPLC analyses show that purity is 97.2%.

The preparation of the TDA types of embodiment 14 crystallization 99% 1 gram of TD grease is dissolved in linl ethyl acetate, resulting solution is slowly added dropwise to the 200ml quickly stirred to be previously cooled in -20 just own protective embankments, separate out solid, filtering, vacuum drying obtains TD crystallizations 0.82g, XRD analysis are shown as TD A types crystallization, and HPLC analyses show that purity is 98.2%.

The physical property characteristic of the TD of embodiment 15 A types crystallization

The full-automatic X-ray diffractometer of A type crystallization D/MAX- IIIC types for the TD that embodiment 11 is obtained（Rigaku Electric Co., Ltd）Survey XRD spectra（Fig. 3), TD A type crystalline characteristics are as follows：

The Θ d value relative intensities of sequence number 2

1 5.92 14.917 48

2 7.66 11.532 37

3 9.04 9.774 77

4 12.40 7.132 32

5 14.00 6.320 53

6 14.70 6.021 33

7 15.46 5.726 68

8 16.44 5.387 46

9 17.00 5.211 46

10 17.84 4.967 58

11 18.28 4.849 100

12 19.08 4.647 49

13 19.48 4.553 41

14 20.04 4.427 36

15 20.60 4.308 35

16 20.86 4.254 34

17 22.04 4.029 32

18 22.82 3.893 33

20 27.66 3.222 28

21 28.58 3.120 26

Use thermal analysis system（DSC2010, TA companies of the U.S.）A types crystallization to TD has carried out differential scanning calorimeter, and is under the rate of heat addition of 10/minute, and it has an endothermic transition peak, and peak value is 100 °C, starting point 97 (see Fig. 4).

Use infrared spectrophotometer（MagNa-IR550, U.S. Buddhist nun high-tensile strength company）Infrared spectrum analysis is carried out using KBr pressed disc methods, the infrared absorption spectroscopy of TD A types crystallization is in the cm " ' of cm -1760 of about 3334 cnT^ of cm -3,164 2979,1659 cm^-1, 1605 cm—！、 1490 cm" 1250 cm— 1142 cm^-1, 980 cm— ¹With 910 cm-¹There is characteristic spectrum belt（See Fig. 5).

Use numeral melting point instrument（WRS-1B Shanghai precision Science and Technology Ltd.）Determine TD A types crystallization, its

Melted between %.2 ~ 97.9 °C.

The preparation of the TD of embodiment 16 Type B crystallization

99% TD (2g) is dissolved in 95% ethanol（In 10ml), room temperature condition is placed in lower 24 hours, obtain TD crystallizations

1.61g, XRD analysis display gained solid crystallizes for TD Type B, and HPLC shows that purity is 98.8%.

The preparation of the TD of embodiment 17 Type B crystallization TD (2g, 95%) is dissolved in acetone（In 15ml), water is instilled in 35 ~ 40 stirrings（In 30ml), 4 °C are cooled to, a small amount of TD Type B crystalline seed is added, crystallized 24 hours, I.4g filtering, vacuum drying obtains white solid.XRD analysis display gained solid crystallizes for TD Type B, and HPLC shows that purity is 97.8%.

The physical property characteristic of the TD of embodiment 18 Type B crystallization

The full-automatic X-ray diffractometer of Type B crystallization D AX-IIIC types for the TD that embodiment 16 is obtained（Rigaku Electric Co., Ltd）Survey its XRD spectrum（See Fig. 6), feature is as follows：

The Θ d value relative intensities of sequence number 2

1 4.38 20.157 53

2 8.84 9.995 50

3 9.46 9.341 15

4 12.02 7.357 19

5 12.32 7.178 20

6 13.34 6.631 14

7 14.08 6.284 17

8 15.94 5.555 34

9 17.24 5.139 23

10 17.88 4.956 15

11 18.48 4.797 16

12 18.88 4.696 33

13 19.94 4.449 100

14 22.40 3.965 55

15 24.18 3.677 43

16 25.82 3.447 15

17 26.16 3.403 22

18 27.02 3.297 72

19 28.54 3.125 26

20 29.12 3.064 10

21 30.26 2.951 11

22 31.68 2.822 42

23 32.94 2.716 11

24 35.58 2.521 8

25 36.42 2.464 13

26 41.18 2.190 8

27 47.08 1.928 6

28 48.26 1.884 6

29 51.92 1.759 7

Thermogravimetic analysis (TGA) system（TGA-7, PE companies of the U.S.）Analysis result shows that TD Type B crystallization has two weightless peaks between 35 ~ 45 °C, altogether weightlessness 6.675%, shows in TD Type B crystallization containing two crystallizations water, its thermogravimetic analysis (TGA)（TG) collection of illustrative plates is as shown in Figure 7.

Use thermal analysis system（DSC2010, TA companies of the U.S.）Type B crystallization to TD has carried out differential scanning calorimeter, under the rate of heat addition of 10'C/ minutes, and it has an endothermic transition peak, and peak value is 55'C, starting point 46'C, and gained differential scanning calorimeter figure is shown in Fig. 8.

Digital Rong points instrument（WRS-1B Shanghai precision Science and Technology Ltd.）Determine TD Type B crystallization 63.2 ~ 64.7 °C it Between melt.

Use infrared spectrophotometer（MagNa-IR550, U.S. Buddhist nun high-tensile strength company）Infrared spectrum analysis is carried out using KBr pressed disc methods, the infrared absorption spectroscopy of TD Type B crystallization is in about 3373 cm'2979 of cm -3203 cm^-1, 1760 cm"¹, 1652 cm— 1605 cm"¹ , 1312 cm"¹, 1250 cm"¹ , 1034 cm— ¹With 965 cm-¹There is characteristic spectrum belt.The representational infrared absorption spectroscopy of TD Type B crystallization is shown in Fig. 9.

The preparation of the unformed solidfied materials of the TD of embodiment 19

99% 1 gram of TD grease is dissolved in 25 ml ethanol, -80 °C or so cryocoagulations, -60 °C of vacuum freeze dryings 24 hours, obtains 0.98 gram of white solid, XRD spectrum as shown in Figure 10, is shown as the unformed solids of TD.

The preparation of the unformed solids of the TD of embodiment 20

99% 1 gram of TD grease is dissolved in lml dichloromethane, resulting solution is slowly added dropwise into quickly stirring

200m Amount are initially cooled in -60 °C of just own protective embankment, continue quick stirring 2 hours after completion of dropwise addition, filter the solid separated out, and vacuum drying obtains solid 0.95g, and XRD analysis are shown as the unformed solids of TD, and HPLC analyses show that purity is 98.5%.The preparation of 21 TD of embodiment-Benexate Hydrochloride

Weigh TD20g, plus the dissolving of 40ml absolute ethyl alcohols；45gp- cyclodextrin is weighed, 567ml water is added and is made into 60 saturated aqueous solutions.TD ethanol solution is instilled in beta-schardinger dextrin saturated aqueous solution, insulated and stirred 30 minutes, stop continuing to stir 4 hours after heating；It is put into refrigerator and freezes 24 hours；Filtering, washs filter cake with absolute ethyl alcohol, is dried under reduced pressure, finely ground, obtains TD 62.5 grams of Benexate Hydrochloride, yield is 96%, and drugloading rate is 30.15%.

The preparation of the TD- Benexate Hydrochlorides of embodiment 22

Weigh TD10g, plus the dissolving of 10ml absolute ethyl alcohols；22.7gp- cyclodextrin is weighed, the mixing of 284ml water is added, room temperature is fully ground into pastel, after low temperature drying, absolute ethyl alcohol cleaning, drying, obtains TD 25 grams of Benexate Hydrochloride, yield is 78%, and drugloading rate is 21.64%.

The preparation of the TD- Benexate Hydrochlorides of embodiment 23

Weigh TD10.02g and 22.7 § beta-schardinger dextrins, in the aqueous solution for being dissolved in 300ml 8 % (v/v) absolute ethyl alcohol, stirring and dissolving and the miillpore filter for passing through 0.45nm, put to refrigerate in liquid nitrogen container and be freeze-dried about 24h again, TD and Benexate Hydrochloride, yield be 98 °/., drugloading rate is 30.5%.

The preparation of the TD fumarates of embodiment 24

The g of TD grease (purity 95%) 5.3 is taken to be dissolved in 30ml methanol, stirring, the fumaric acid methanol solution 10ml of the g containing U6 is slowly added dropwise into this solution simultaneously, it is stirred continuously under 25 °C of constant temperature 1 hour, filtering removes insoluble matter, is positioned over suction filtration after 0 ~ 4 °C, 5 hours, obtain 4.8 grams of the solid of white, 119 °C of fusing point.

¾NMR (DMSO-d₆): 8.13(lH,s,H-8)、 8.03(lH,s,H-2)、 7.15(2H,s,NH₂), 6.63 (2H, s, fumaric acid H-2, H-3), 5.54 (4H, m, CH₂OP) 、 4,21(2H,ddd, J=4,l,4,4,3,4.8)、 3.94(3H,m,H-4,H-4')、 1.15(18H,d,J=3.2,CH₃)、 1.62(3H,d,J=6,H-3

A unimodal characteristic peak for fumaric acid H-2, H-3 at 6.63 on ifiNMR collection of illustrative plates, by integration understand TD and fumaric acid into salt ratio be 1: 1.' HNMR collection of illustrative plates is as shown in figure 11.

IR collection of illustrative plates is as shown in figure 12.

As shown in figure 13, feature is as follows for XRD spectrum：

The Θ d value relative intensities of sequence number 2

1 4.72 18.706 42

2 10.60 8.339 6

3 13.04 6.783 7

4 14.48 6.112 32

5 17.46 5.075 9

6 19.44 4.562 48

7 20.10 4.414 8

8 21.44 4.141 8

9 21.96 4.044 8

' 10 23.54 3.776 8

11 24.40 3.645 100

12 24.98 3.561 10

13 26.42 3.370 7

14 27.36 3.257 8

15 28.48 3.131 7

16 29.42 3.033 44

17 29.90 2.985 9

18 33.24 2.693 6

Shang：^ 34.52 2.596 12

20 39.66 2.270 4

21 50.26 1.813 3

The preparation of the TD fumarates of embodiment 25

The pure g of TD grease 5.15 is dissolved in 30ml acetone, and slow be added dropwise into this solution contains 1.16 while stirring_!gFumaric acid methanol solution 10ml, it is stirred continuously under 25 °C of constant temperature 1 hour, filtering removes insoluble matter, rotary evaporation remove solvent after by residual solids 45 °C be dissolved in 20 ml ethyl acetate, 0 ~ 4 °C of suction filtration after 12 hours is positioned over, 5.5 grams of the solid of TD fumarates white is obtained.119 °C of fusing point.

The preparation of the TD oxalates of embodiment 26

TD grease 5.15g is taken to be dissolved in 30ml ethyl acetate, stirring, it is simultaneously slow that the oxalic acid ethanol solution containing 0.9 g is added dropwise into this solution, it is stirred continuously under 45 constant temperature, insoluble matter will be filtered to remove with about 20 minutes after oxalic acid ethanol solution completion of dropping, room temperature is gradually decreased to, continues suction filtration after the small inch of stir about 5,4.6 grams of the solid of TD Oxalateses white is obtained.153-154 °C of fusing point.

¹HNMR(DMSO-d6):8.15 (lH, s, H-8), 8.05 (lH, s, H-2), 7.29 (2H, s, NH₂)、

5.54 (4H, m, C OP), 4.22 (2H, ddd, J=0.4,14.4,35.6, H-l, H-l, H-2), 3.95 (3H, m, H-4, H-4'), U5 (18H, d, J=2.8, C), 1.08 (3H, d, J=6, H- 3), ^NMR collection of illustrative plates it is as shown in figure 14. As shown in figure 15, XRD spectrum is as shown in figure 16 for IR collection of illustrative plates.

The preparation of the TD salicylates of embodiment 27

Take TD grease or unformed solidfied material or crystal 5.15_gIt is dissolved in 30ml ethyl acetate, stirring, it is simultaneously slow that the salicylic acid ethanol solution containing 1.76 g is added dropwise into this solution, it is stirred continuously under 45 ' C constant temperature, with about 20 minutes by salicylic acid ethanol solution completion of dropping, insoluble matter is filtered to remove, room temperature is gradually decreased to, continuation stir about obtains off-white color solid after 8 hours is TD salicylates.Fusing point 88V, IR collection of illustrative plates is as shown in figure 17.

The preparation of the TD oleanolic acid salts of embodiment 28

Take 99% g of TD crystal 5.15 to be dissolved in 30ml dichloromethane, 4.5g oleanolic acid is added in this backward solution in 100ml ethanol:Dichloromethane（1 :1) solution in, is stirred continuously after 120 minutes under 50 °C of constant temperature and solvent in mixture is removed in vacuum, and it is TD oleanolic acid salts to obtain linen solid, and fusing point 242'C (is decomposed）, IR collection of illustrative plates is as shown in figure 18.

The preparation of the TD aspartates of embodiment 29

Take 99% g of TD crystal 1.0 to be dissolved in 10ml ethanol, stir, while slow be added dropwise into this solution contains 0.266_gAspartic acid（Preferred L-Aspartic acid）The aqueous solution, (is stirred continuously under TC constant temperature 4, with about 20 minutes by aspartic acid aqueous solution completion of dropping, continues to be gradually decreased to room temperature after stirring 150 minutes at this temperature, vacuum freeze drying obtains the solid of off-white color, 163 °C of fusing point.

The preparation of the TD taurates of embodiment 30

99% g of TD crystal 1.0 is taken to be dissolved in 10ml ethanol; the taurine aqueous isopropanol containing 0.25 g is added dropwise into this solution; the solid that solvent in mixture obtains off-white color, 172 °C of fusing point is removed in vacuum in stir about under 45'C constant temperature after 120 minutes.

The preparation of the TD hydrochlorides of embodiment 31

99% TD crystal 1.03g is taken to be dissolved in lOmlTHF, 1M hydrogen chloride THF solution 2.2ml is added dropwise at 0 °C, continue stir about after completion of dropping and -20 °C are placed after 120 minutes overnight, 0.95 gram of the white solid of filtering, 192 °C of fusing point (is decomposed）.

The preparation of the TD Hemisulphates of embodiment 32

Take 99% TD crystal 1.03g to be dissolved in lOmlTHF, stir, be added dropwise at 0 °C after 1M methanolic solution 2.2ml, completion of dropping and continue stir about 120 minutes, vacuum freeze drying obtains the solid of white.

The preparation of the TD tosilate of embodiment 33

Take 99% TO crystal 1.03g to be dissolved in lOmlTHF, stir, (continuing stir about 120 minutes after TC dropwise additions 1M p-methyl benzenesulfonic acid methanol solution 2.2ml, completion of dropping, the foaming solid for obtaining white is removed in vacuum.

The TD of embodiment 34 A types crystallize the preparation of tablet

2S Prescription（Based on 1000）：The TD crystallization of A types 30g, lactose 200g, carboxyrnethyl starch sodium 2g, PVP（K30) 15g, magnesium stearate 0.4_g, talcum powder 1.2g.

Preparation method：TD A types are crystallized, lactose, carboxyrnethyl starch sodium, PVP（00), magnesium stearate, talcum powder crosses SO mesh sieves respectively, standby.Then by the tenofovir disoproxil of prescription full dose, lactose, carboxyrnethyl starch sodium, PVP (K30), the magnesium stearate of recipe quantity 50%, talcum powder is sufficiently mixed with equally addition and hooked, and the granulation of 18 mesh sieves is crossed with Drygranulatemachine；Remaining stearic acid mirror is added, talcum powder is sufficiently mixed uniform, tabletting, produces every tablet of tablet containing 30 milligrams of TD.The TD of embodiment 35 Α types crystallize the preparation of tablet

Prescription（Based on 1000）：The TD crystallization of A types 10g, starch 100g, carboxyrnethyl starch sodium 2g, PVP（K30) 10g, magnesium stearate 0.4g, talcum powder 1.2g, magnesium carbonate 2_g。

Preparation method：TD A types are crystallized, starch, carboxyrnethyl starch sodium, PVP（K30), magnesium stearate, talcum powder and calcium carbonate, 80 mesh sieves are crossed respectively, then the TD of recipe quantity A types are crystallized, starch, carboxyrnethyl starch sodium, PVP (30) and magnesium carbonate mixing, add appropriate water and prepare softwood, sieving granulation determines content and moisture after drying, ' add magnesium stearate and talcum powder and be sufficiently mixed uniformly, tabletting is produced.

The preparation of the tablet of the TD fumarates of embodiment 36

Prescription（Based on 1000）：TD fumarate 50g, starch 1000g, VBT（L- stone hydrochlorates）200g, carboxyrnethyl starch sodium 20g, PVP（K30) 10g, magnesium stearate 2g, talcum powder 5g.

Preparation method：Auxiliary material in TD fumarates and prescription is crossed into 80 mesh sieves respectively, then by the TD fumarates of recipe quantity, starch, VBT（L-TARTARIC ACID salt）, carboxyrnethyl starch sodium, PVP（K30) mix, add appropriate water and prepare softwood, sieving granulation, for determining content and moisture after doing, adds magnesium stearate and talcum powder is sufficiently mixed uniformly, tabletting is produced.The TD of embodiment 37 A types crystallize the preparation of capsule

Prescription（Based on 1000）：The TD crystallization of A types 30g, pregelatinized starch 200g, talcum powder 2g.

Preparation method：Take main ingredient and each auxiliary material to crush 100 mesh sieves after drying respectively standby, main ingredient and each auxiliary material are measured by prescription, it is well mixed by equally addition；Determine mixed-powder content, moisture；Powder directly filling is produced.

The preparation of the TD fumarate capsules of embodiment 38

Prescription（Based on 1000）：TD fumarate 50g, pregelatinized starch 400g, VBT（L-TARTARIC ACID salt）100g, talcum powder 10g.

Preparation method：Take main ingredient and each auxiliary material to crush 100 mesh sieves after drying respectively standby, main ingredient and each auxiliary material are measured by prescription, it is well mixed by equally addition；The granulation of 18 mesh sieves is crossed with thousand formula comminutors, mixed-powder content, moisture is determined；It is particles filled to produce.

The TD A types of embodiment 39 crystallize the preparation of dispersible tablet

Prescription（Based on 1000）：TD A types crystallization 10_g, pregelatinized starch 20g, microcrystalline cellulose 60g, lactose 20g, carboxyrnethyl starch sodium 25g, 12 protective embankment base sodium sulphate lg, magnesium stearate lg.

Preparation method：Recipe quantity TD A types were crystallized into 100 mesh sieves, recipe quantity pregelatinized starch is separately taken, microcrystalline cellulose, lactose, carboxyrnethyl starch sodium, lauryl sodium sulfate, magnesium stearate crosses 60 mesh sieves, were well mixed.Main ingredient and each auxiliary material are measured by prescription, is well mixed by with waiting fl addition, determines content, direct powder compression is produced.The disintegration time of obtained tablet is less than 1 minute.

The TD- beta cyclodextrin clathrate injection powder pins of embodiment 40

Prescription：

TD- beta cyclodextrin clathrates（Carrying drug ratio 30%) 10 grams

5.5 grams of trisodium citrate

500 grams of mannitol

Water for injection adds to 1000ml

Be made altogether 1000 bottles '

Preparation technology：

The sodium citrate of recipe quantity is taken, is dissolved in appropriate water for injection, by the TD- beta cyclodextrin clathrates of recipe quantity（Carrying drug ratio 30%) add, it is stirred to dissolve, plus water for injection about 900ml, the mannitol of recipe quantity is being added, Jiao, which is mixed, makes dissolving：PH5.5 or so is adjusted with 0.1mol/L citric acid solution, is added to the full amount of water for injection, 0.03% (m/v) needle-use activated carbon is added and stirs 30 minutes, through 0.22 μ π ι miillpore filter malleation aseptic filtrations；After the inspection of semifinished product is qualified, in the aseptic subpackaged vial with after washing and sterilizing, per bottled amount hnl;Through frozen drying about 24 hours, sealing was produced.Packed after product inspection is qualified.

The TD fumarate parenteral solutions of embodiment 41, injection for intravenous.

Prescription：

3.3 grams of TD fumarates

9.0 grams of sodium chloride

Appropriate water for injection

Full dose 1000ml

1000 bottles are made altogether

TD fumarates, the sodium chloride of preparation technology-weigh recipe quantity, plus water for injection 900ml, it is heated to 80 °C of dissolvings, again with 0.1mol/L lemons acid for adjusting pH to 4.0 5.0, add to the full amount of water for injection, plus activated carbon 0.01%w/v, 15min is stirred, by sand rod decarburization, then through 0.45 μ ι η filtering with microporous membrane, filtrate is filling in 100ml glass infusion bottles, mylar is put, plug, gland is covered, 115 °C of flowing steam sterilizations 30 minutes, lamp inspection, packaging are produced.

Claims (1)

1. Claim

   1. the application of compound (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine or its physiologically acceptable derivative in the scorching virus of anti-hepatitis B shown in formula (I)

   The crystallization of one kind (R) -9- 2. [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine, it is characterised in that：Its X-ray powder diffraction spectrum crystal face represents generally that in 9.774 A, 6.32 A 5.726 A, 4.967 A, 4.849 A have peak away from d values.

   3. the crystallization described in claim 2, it is characterised in that：Its X-ray powder diffraction spectrum crystal face represents generally there is peak in 14.917 people, 9.774 A, 6.32 A, 5.726 A, the A of 5.387 As 5,211,4,967 A, 4.849 A, 4.647 A, 4.553 A, 3.817 A away from d values.

   4. described in claim 2 (crystallization of R 9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine, it is characterised in that：Its Differential scanning calorimetry collection of illustrative plates has maximum endothermic peak at 100 °C or so.

   5. one kind (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyl] Xian Piao Ling of 2- crystallization, described crystallization, it is characterised in that：Its X-ray powder diffraction spectrum crystal face represents generally that in 20.157 people, 9.995 A, 4.449 A 3.965 k, 3.297 A have peak away from d values.

   6. the crystallization described in claim 5, it is characterised in that：Its X-ray powder diffraction spectrum crystal face represents generally that in 20.157 A, the A of 9.995 A. 5.555,4.696 A, 4.449 A, 3.965 A, the A of 3.677 A. 3.297 3.125 people, 2.822 A have peak away from d values.

   7. the crystallization of (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine described in claim 5, it is characterised in that：Its Differential scanning calorimetry collection of illustrative plates has maximum endothermic peak at 55 °C or so.

   8.-kind of (R) -9- [2- pairs (；Pivaloyloxymethyl；) phosphonylmethoxypropyl] and adenine unformed solidfied material, it is characterised in that：Wherein the content of TD unformed shapes is more than 70%.

   9. the salt of (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] fast cry of certain animals of gland as shown in formula (II),

   (II)

   Wherein a is acid and (mol ratio of R 9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine, a is between 1 ~ 5：HA is acid.

   10. (R) -9- [2- double] pivaloyloxymethyl described in claim 9) phosphonylmethoxypropyl] adenine salt, wherein HA be 5 sulfuric acid, phosphoric acid, nitric acid, hydrochloric acid, hydroiodic acid, hydrobromic acid, hydrofluoric acid, taurine, benzene sulfonic acid, p-methyl benzenesulfonic acid, α-naphthalenesulfonicacid, beta-naphthalenesulfonic-acid,（S)-camphorsulfonic acid, methanesulfonic acid, ethyl sulfonic acid, positive propane sulfonic acid, isopropyl sulfonic acid, positive fourth sulfonic acid, Zhong Ding sulfonic acid, isobutyl sulfonic acid, tertiary fourth sulfonic acid, penta sulfonic acid, own sulfonic acid, acetic acid, glycolic, lactic acid, pyruvic acid, malonic acid, glutaric acid, tartaric acid, citric acid, fumaric acid, butanedioic acid, malic acid, maleic acid, oxalic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, tussol, cinnamic acid, mandelic acid, the poplar of water 10 acid, 1- phenoxy benzoic acids, nicotinic acid, pantothenic acid, aspartic acid, glutamic acid, valine, ascorbic acid, oleanolic acid, malol, ursolic acid, glycyrrhizic acid, enoxolone, red sage root acid, forulic acid, glucuronic acid, gluconic acid or levulinic acid.

   11. (R) -9- [2- double] pivaloyloxymethyl described in claim 9) phosphonylmethoxypropyl] adenine salt, wherein HA is fumaric acid, oxalic acid, salicylic acid, oleanolic acid and aspartic acid.

   The fumarate of 15 12. (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine as shown in formula (Π Ι)：

   (III)

   13. the crystallization of 0-9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine fumarate described in claim 12, it is characterised in that：Its X-ray powder diffraction spectrum crystal face represents generally there is peak in 18.706A, 6.112A, 4.562A, 3.645A, 3.561 A, 3.033A, 2.596 people away from d values.

   The cyclodextrin inclusion compound of 20 14. (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine, it is characterised in that：(R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2- " mol ratio of Xian Piao Ling and cyclodextrin is 1.1 ~ 1 Λ 0.

   15. (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of the 2-] purposes of adenine and its derivative in antiviral of the solid state described in claim 2,5,8,9,12 or 14.

   16. a kind of composition, (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyl] Xian Piao Ling of 2- or its physiologically acceptable derivative and pharmaceutical carrier containing solid state.

   17. (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of the 2-] adenines or its physiologically acceptable derivative described in claim 16 are the crystallization of A types, Type B crystallization, unformed solidfied material, salt or the cyclodextrin inclusion compound of (R) -9- [double (pivaloyloxymethyl) phosphonylmethoxypropyls of 2-] adenine.

   18. the composition described in claim 16, it is characterised in that：Described composition also contains VBT or its salt.

   19. the composition described in claim 16, it is characterised in that：Described composition also contains basic medicinally carrier.