KR100971486B1 - Adefovir dipivoxil malonate and pharmaceutical composition including the same - Google Patents

Abstract

PURPOSE: An adefovir dipivoxil malonate is provided to ensure stability, non-absorption property, and solubility and to use in pharmaceutical composition for treating HIV and hepatitis type B infection. CONSTITUTION: An adefovir dipivoxil malonate is crystalline and is denoted by chemical formula I. The adefovir dipivoxil malonate has diffraction angle(2θ) having 10% or more of I/I0(I: peak intensity in the diffraction angle, I0: maximum peak intensity) is 8.1±0.2, 12.1±0.2, 16.1±0.2, 17.1±0.2, 19.1±0.2, 19.5±0.2, 20.9±0.2, 22.6±0.2, 24.6±0.2, 26.6±0.2, and 32.9±0.2. A pharmaceutical composition for treating HIV or hepatitis type B infection contains the adefovir dipivoxil malonate and pharmaceutically acceptable carrier.

Description

Adefovir dipivoxil malonate and pharmaceutical composition including the same}

The present invention relates to adefovir dipoxyl malonate and pharmaceutical compositions comprising the same. More specifically, the present invention relates to adefovir difficyl malonate and a pharmaceutical composition comprising the same, which are excellent in stability, non-hygroscopicity and solubility, which can be effectively used in pharmaceutical compositions for the treatment of HIV and hepatitis B virus infection. .

Adefovir difixyl (9- [2-[[bis [(pivaloyloxy) -methoxy] phosphonyl] methoxy] ethyl] adenine) represented by the following formula (II) is a nucleotide reverse transcriptase inhibitor, And excellent in vivo antiviral activity against hepatitis B (HBV) (U.S. Pat. No. 5,731,591, J. Infect. Dis., 176 (2): 406, 1997 and J. Med. Chem., 37 : 1857-1864, 1994).

Adefovir difficile was developed to improve the low oral absorption rate of 9- (phosphonylmethoxyethyl) adenine (Adefovir; US Patent No. 4808716), which was previously developed as an antiviral agent. Adefovir difficile, developed as a prodrug of adefovir, is a reverse transcriptase inhibitor that has been developed for the treatment of HIV, and its use as a drug for HIV has been discontinued due to kidney toxicity. Currently, it has been developed and used as a treatment for adult patients with chronic hepatitis B who have evidence of active virus proliferation and have elevated blood ALT (aspartate aminotransferase) or AST (aspartate aminotransferase). Adefovir difficile is a kind of purine-type nucleoside that can be used orally. Like other nucleosides, it acts as a chain terminator, DNA polymerase inhibitory effect, and immune system function enhancement. It has a role in slowing the development of chronic hepatitis. It has also been shown to have an inhibitory effect on lamivudine-resistant HBV mutants.

However, adefovir difficile chamber has not only low stability against heat and moisture, but also has high hygroscopicity and low solubility in water. In general, increasing the stability of the drug against heat and water can provide a better pharmaceutical composition because the active ingredient decomposes in the process of manufacturing the drug into tablets, thereby lowering the possibility of causing impurities and allowing long-term storage. . In addition to the requirement that the active ingredients used in the pharmaceutical compositions have good thermal stability and no moisture hygroscopicity, they must also meet the conditions of high solubility in water or aqueous solutions over a wide pH range. This is because the solubility of the active ingredient in water or aqueous solution has a great effect on the dissolution rate of the pharmaceutical composition, and consequently, the bioavailability of the active ingredient is greatly affected. For this reason, there has been a need for the development of new crystalline forms or new acid addition salts that can be expected to be more stable and higher bioavailability.

On the other hand, U.S. Patent No. 6645340 and Republic of Korea Patent No. 618663 have anhydrous crystalline form (crystalline form 1), hydrated form (crystalline form 2) and solvate with crystalline form (crystalline form 3) of adefovir difficile. It is disclosed that it has good melting point and / or flow rate or capacity density characteristics to facilitate ignition. In addition, US Patent Registration No. 7417036 and Korean Patent Publication No. 2005-0086415 disclose a new crystalline form of adefovir die fibsil. In addition, U.S. Patent No. 6645340 and U.S. Patent No. 618663 introduce new acid addition salts of adefovir difficile. Specifically, new acid addition salts include fumarate, hemisulfate, hydrobromide, hydrochloride, nitrate, Methanesulfonate, ethanesulfonate, β-naphthalenesulfonate, α-naphthalenesulfonate, (S) -camposulfonate, succinate, maleate, ascorbate and nicotinate, with their respective melting points and / or crystalline structures Is disclosed. In addition, Chinese Unexamined Patent Publication No. 1528766 discloses adefovir diepoxyl oxalate as a novel acid addition salt of adefovir diepsil.

However, the above-mentioned patent documents do not describe the contents related to the stability and hygroscopicity of the crystalline form of the adefovir difficile acid and the heat and moisture of acid addition salts. On the contrary, since the ester structure of adefovir difficile has a property of being easily hydrolyzed by an acidic catalyst or a basic catalyst, it is judged that acid addition salt has a high possibility of negatively affecting the stability of the adefovir difficile. come. According to U.S. Patent No. 6635278 and Korean Patent Registration No. 624214, the stability of the formulation is reported to be superior when the adefovir difficile and the basic excipient are used together without the basic excipient. .

The present inventors have diligently researched to develop adefovir difficile compounds having excellent formulation properties such as stability against heat and moisture, and non-hygroscopicity. Surprisingly, adefovir difficyl malonate is not only highly stable against heat and moisture, but also hygroscopic This low and high solubility in water was found and the present invention was completed.

It is therefore an object of the present invention to provide adefovir difixyl malonate excellent in stability, non-hygroscopicity and solubility.

Another object of the present invention is to provide a pharmaceutical composition for the treatment of HIV or hepatitis B virus infection comprising adefovir difficile malonate.

The present invention relates to adefovir dipoxyl malonate represented by the following general formula (I).

The adefovir difixyl malonate of the present invention is preferably crystalline adefovir difixyl malonate, more preferably I / I ₀ (I: intensity of peak at each diffraction angle, I in X-ray diffraction analysis) ₀ : the diffraction angle (2θ) with the intensity of the largest peak) of 10% or more is 8.1 ± 0.2, 12.1 ± 0.2, 16.1 ± 0.2, 17.1 ± 0.2, 19.1 ± 0.2, 19.5 ± 0.2, 20.9 ± 0.2, 22.6 ± It is crystalline adefovir difixyl malonate (hereinafter referred to as 'crystalline Form I'), which is 0.2, 24.6 ± 0.2, 26.6 ± 0.2, 32.9 ± 0.2.

Alternatively, the adefovir difficyl malonate of the present invention preferably has an I / I ₀ (I: intensity of peak at each diffraction angle, I ₀ : intensity of largest peak) in X-ray diffraction analysis. The diffraction angle (2θ) with a% or more is 11.1 ± 0.2, 14.6 ± 0.2, 16.5 ± 0.2, 17.2 ± 0.2, 17.5 ± 0.2, 18.3 ± 0.2, 19.0 ± 0.2, 19.5 ± 0.2, 19.8 ± 0.2, 20.5 ± 0.2, 20.8 ± 0.2, 21.7 ± 0.2, 22.1 ± 0.2, 22.6 ± 0.2, 23.0 ± 0.2, 24.8 ± 0.2, 29.6 ± 0.2, crystalline adefovir difficyl malonate (hereinafter referred to as 'crystalline Form II') )to be.

The adefovir difficile malonate according to the present invention not only solved the problem of heat and moisture instability and hygroscopicity caused by adefovir dificil and its other acid addition salts, but also by using malonic acid which is one of the safest organic acids. Pharmaceutically very advantageous advantages. The malonic acid used in the present invention is not only very safe with a value of 4000 mg / Kg of LD ₅₀ (50% lethal dose in mice) [Handbook of Pharmaceutical Salts, p 293 (2008)], molecular weight 104.03 It is an organic acid which is very advantageous for the preparation of acid addition salts because it is relatively small as g / mol.

In addition, the adefovir difficile malonate according to the present invention is expected to increase the solubility at various pH conditions such as stomach, intestine, blood, etc., thereby increasing the absorption and utilization of the active ingredient in the body.

The adefovir dipoxyl malonate represented by the formula (I) according to the present invention can be prepared by reacting the adefovir dipoxyl represented by the following formula (II) with malonic acid represented by the following formula (III).

Hereinafter, a method for preparing adefovir dipoxyl malonate according to the present invention will be described in more detail.

It is preferable that the adefovir difixyl malonate of the present invention is prepared by dissolving adefovir difixyl and malonic acid in an organic solvent and stirring. It can be prepared by dissolving adefovir dificil and malonic acid in an organic solvent, respectively, or mixing adefovir dificil and malonic acid together in an organic solvent. The amount of malonic acid to be used is preferably about 1 equivalent to the adefovir difficile.

In the method for preparing adefovir dipoxyl malonate according to the present invention, the step of reacting the adefovir dipoxyl represented by Formula (II) with malonic acid represented by Formula (III) in an organic solvent,

(i) concentrating the reaction solution under reduced pressure, adding a precipitated solvent to the obtained residue, stirring and filtering the obtained solid;

(ii) lowering the temperature of the reaction solution and stirring, and filtering the obtained solid;

(iii) adding a precipitating solvent to the reaction solution, and stirring, and filtering the obtained solid.

Examples of the organic solvent include alcohols such as methanol, ethanol, isopropanol, 1-butanol and 1-hexanol, ethers such as tetrahydrofuran, dioxane and isopropyl ether, nitriles such as acetonitrile and acetone And ketones such as 2-butanone and esters such as ethyl acetate and isopropyl acetate can be used.

Examples of the precipitation solvent include alcohols such as isopropanol, 1-butanol, and 1-hexanol, ethers such as tetrahydrofuran, dioxane, and isopropyl ether, nitriles such as acetonitrile, acetone, and 2-view Ketones such as tanone, hydrocarbons such as n-pentane, n-hexane, aromatic hydrocarbons such as benzene, toluene and xylene, esters such as ethyl acetate and isopropyl acetate, and dichloromethane, chloroform, 1,2 One or more selected from chlorinated hydrocarbons such as dichloroethane can be used.

The reaction time is preferably 1 to 5 hours, the reaction temperature is preferably 20 to 60 ℃. The method for preparing adefovir difixyl malonate of the present invention may further include the step of filtering and then washing and drying the obtained solid.

On the other hand, the present invention provides a pharmaceutical composition comprising adefovir difficile malonate together with a pharmaceutically acceptable carrier, specifically for the treatment of HIV or hepatitis B virus infection, in particular chronic hepatitis B virus infection. It relates to a pharmaceutical composition.

The pharmaceutical composition of the present invention may include other bioactive substances in addition to adefovir dipoxyl malonate.

The pharmaceutical composition according to the present invention may be formulated in various forms such as tablets, capsules, granules, powders, emulsions, suspensions, syrups and the like. The various types of pharmaceutical compositions include excipients, fillers, extenders, binders, disintegrators, lubricants, preservatives, antioxidants, isotonic agents, buffers, coatings, sweeteners, solubilizers, bases, dispersants It can be prepared by known techniques using pharmaceutically acceptable carriers commonly used in each formulation, such as wetting agents, suspending agents, stabilizers, colorants, fragrances.

The content of the adefovir difficyl malonate of the present invention in the preparation of the medicament depends on the form of the medicament, but is preferably in a concentration of 0.01 to 10% by weight, more preferably 0.01 to 1% by weight.

The dosage of the pharmaceutical composition of the present invention varies in a wide range depending on the type of mammal including the person to be treated, route of administration, weight, sex, age, degree of disease, judgment of the doctor, and the like. In general, in the case of oral administration, 0.05 to 0.5 mg of the active ingredient may be administered per kg body weight per day. The above-described daily dose may be used at one time or dividedly depending on the extent of the disease, the judgment of the doctor and the like.

Adefovir difficile malonate according to the present invention is a pharmaceutical composition for the treatment of HIV and hepatitis B virus infection by improving the stability of heat and water, lower hygroscopicity, and low solubility in water of adefovir difficile malpilate Can be used effectively.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it is obvious to those skilled in the art that the scope of the present invention is not limited to these examples.

Example 1 Preparation of Adefovir Difixyl Malonate (crystalline Form I)

5.01 g (10.0 mmol) of anhydrous adefovir difficile and 1.04 g (10.0 mmol) of malonic acid were completely dissolved in 50 ml of methanol and stirred at 20-25 ° C. for 2 hours. The reaction solution was concentrated by distillation under reduced pressure, and 35 ml of acetone was added to the residue, followed by stirring at 20 to 25 ° C for 3 hours. The resulting white crystalline solid was filtered, washed with 5 ml of cooled acetone, and dried under vacuum at 50 ° C. for 24 hours to give 4.40 g of adefovir dipoxyl malonate (crystalline Form I). The yield was 72.7%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile malonate were carried out and the results are shown in FIGS. 1 and 2, respectively.

Melting Point: 108 ~ 110 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.09 (s, 1 H), 8.03 (s, 1 H), 7.19 (brs, 2 H), 5.52 (s, 2 H), 5.49 (s , 2 H), 4.29 to 4.27 (m, 2 H), 3.92 (d, 2 H, J = 7.6 Hz), 3.86 to 3.84 (m, 2 H), 3.19 (s, 2 H), 1.09 (s, 18 H)

Example 2: Preparation of Adefovir Difixyl Malonate (crystalline Form II)

5.01 g (10 mmol) of anhydrous adefovir difficile and 1.04 g (10 mmol) of malonic acid were completely dissolved in 30 mL of isopropanol. After 200 ml of isopropyl ether was added dropwise to the reaction solution, the mixture was stirred at 20 to 25 ° C. for 5 hours, and further stirred at 0 ° C. for 1 hour. The resulting white crystalline solid was filtered, washed with 5 ml of cooled isopropyl ether, and dried under vacuum at 50 ° C. for 24 hours to give 5.45 g of adefovir dipoxyl malonate (crystalline Form II). The yield was 90.1%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile malonate were carried out and the results are shown in FIGS. 3 and 4, respectively.

Melting Point: 104 ~ 106 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.09 (s, 1 H), 8.03 (s, 1 H), 7.19 (brs, 2 H), 5.52 (s, 2 H), 5.49 (s , 2 H), 4.29 to 4.27 (m, 2 H), 3.92 (d, 2 H, J = 7.6 Hz), 3.86 to 3.84 (m, 2 H), 3.19 (s, 2 H), 1.09 (s, 18 H)

Comparative Example 1: Preparation of Adefovir Difisil Glutarate

5.01 g (10.0 mmol) of anhydrous adefovir difficile acid and 1.32 g (10.0 mmol) of glutaric acid were completely dissolved in 50 mL of methanol, followed by stirring at 20 to 25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, 20 ml of acetone and 120 ml of isopropyl ether were added to the residue, and the mixture was heated to 50 ° C. to completely dissolve it. 60 mL of the solvent was removed under reduced pressure and stirred at 20-25 ° C. for 3 hours. The resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to give 5.44 g of adefovir dipoxyl glutarate. The yield was 86.0%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile glutarate were performed and the results are shown in FIGS. 5 and 6, respectively.

Melting Point: 86 ~ 88 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.09 (s, 1 H), 8.03 (s, 1 H), 7.18 (brs, 2 H), 5.53 (s, 2 H), 5.50 (s , 2H), 4.29 (t, 2H, J = 5.2 Hz), 3.93 (d, 2H, J = 8.0 Hz), 3.86 (t, 2H, J = 5.2 Hz), 2.20 (t, 4H , J = 7.2 Hz), 1.70 to 1.63 (m, 2 H), 1.09 (s, 18 H)

Comparative Example 2: Preparation of Adefovir Difixyl L-Mandelate

2.50 g (5.0 mmol) of anhydrous adefovir difficile acid and 0.76 g (5.0 mmol) of L-mandelic acid were completely dissolved in 50 mL of methanol, followed by stirring at 20 to 25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, 15 ml of isopropanol and 100 ml of isopropyl ether were added to the residue, followed by stirring at room temperature for 12 hours. The resulting white crystalline solid was filtered off and dried under vacuum at 50 ° C. for 24 hours to give 3.10 g of adefovir difficile L-mandelate. Yield 95.0%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile L-mandelate were performed and the results are shown in FIGS. 7 and 8, respectively.

Melting Point: 100 ~ 108 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.09 (s, 1H), 8.03 (s, 1H), 7.40-7.23 (m, 5H), 7.17 (brs, 2H) , 5.53 (s, 2H), 5.50 (s, 2H), 4.98 (s, 1H), 4.29 (t, 2H, J = 5.2 Hz), 3.93 (d, 2H, J = 7.6 Hz) , 3.86 (t, 2H, J = 5.2 Hz), 1.10 (s, 18H)

Comparative Example 3: Preparation of Adefovir Difixyl Salicylate

2.50 g (5.0 mmol) of anhydrous adefovir difficile and 0.70 g (5.0 mmol) of salicylic acid were completely dissolved in 50 mL of methanol, and then stirred at 20 to 25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, 10 ml of acetone and 50 ml of isopropyl ether were added to the residue, followed by stirring at room temperature for 12 hours. The resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to give 2.41 g of adefovir dipoxyl salicylate. The yield was 75.5%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile salicylate were carried out and the results are shown in FIGS. 9 and 10, respectively.

Melting Point: 90 ~ 96 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.10 (s, 1H), 8.04 (s, 1H), 7.76-7.77 (m, 1H), 7.48-7.44 (m, 1H) , 7.21 (brs, 2 H), 6.91 to 6.85 (m, 2 H), 5.53 (s, 2 H), 5.50 (s, 2 H), 4.29 (t, 2 H, J = 5.2 Hz), 3.92 ( d, 2H, J = 7.6 Hz), 3.86 (t, 2H, J = 5.2 Hz), 1.10 (s, 18H)

Comparative Example 4: Preparation of Adefovir Dificil Orotate

2.50 g (5.0 mmol) of anhydrous adefovir difficile and 0.75 g (5.0 mmol) of orotic acid monohydrate were added to 50 mL of methanol, followed by reflux stirring for 1 hour to completely dissolve. The reaction solution was slowly cooled to 20-25 ° C. and stirred for 3 hours. The resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to yield 2 g of adefovir dipoxyl orotate. The yield was 77.8%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir dipoxyl orotate were carried out and the results are shown in FIGS. 11 and 12, respectively.

Melting Point: 186 ~ 190 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 11.28 (s, 1 H), 10.76 (brs, 1H), 8.14 (s, 1 H), 8.08 (s, 1 H), 7.45 (brs, 2 H), 5.95 (d, 1 H, J = 1.6 Hz), 5.53 (s, 2 H), 5.50 (s, 2 H), 4.30 (t, 2 H, J = 5.2 Hz), 3.93 (d, 2 H, J = 7.6 Hz), 3.86 (t, 2 H, J = 5.2 Hz), 1.08 (s, 18 H)

Comparative Example 5: Preparation of Adefovir Dicarboxyl Gentiate

2.50 g (5.0 mmol) of anhydrous adefovir difficile acid and 0.77 g (5.0 mmol) of gentic acid were completely dissolved in 50 mL of methanol, followed by stirring at 20 to 25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, 10 ml of isopropanol and 90 ml of isopropyl ether were added to the residue, and the mixture was heated to 50 ° C to completely dissolve it. The reaction solution was stirred at room temperature for 12 hours, and the resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to obtain 2.14 g of adefovir dicarboxyl genitate. The yield was 65.2%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile gentiate were carried out and the results are shown in FIGS. 13 and 14, respectively.

Melting Point: 122 ~ 124 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.11 (s, 1H), 8.04 (s, 1H), 7.24 (brs, 2H), 7.11 (d, 1H, J = 2.8 Hz ), 6.93 to 6.90 (m, 1 H), 6.74 (d, 1 H, J = 8.8 Hz), 5.53 (s, 2 H), 5.50 (s, 2 H), 4.29 (t, 2 H, J = 5.2 Hz), 3.92 (d, 2H, J = 7.6 Hz), 3.86 (t, 2H, J = 5.2 Hz), 1.05 (s, 18H)

Comparative Example 6: Preparation of Adefovir Difisil DL-Malate

2.50 g (5.0 mmol) of anhydrous adefovir difficile acid and 0.67 g (5.0 mmol) of DL-malic acid were completely dissolved in 50 mL of methanol, followed by stirring at 20 to 25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, 50 ml of acetone was added to the residue, and the mixture was heated to 50 ° C to completely dissolve it. The reaction solution was slowly cooled to 20-25 ° C. and stirred at the same temperature for 12 hours. The resulting white crystalline solid was filtered and then dried under vacuum at 50 ° C. for 24 hours to give 2.94 g of adefovir difficile DL-maleate. The yield was 92.4%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile DL-maleate were carried out and the results are shown in FIGS. 15 and 16, respectively.

  Melting Point: 98 ~ 104 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.08 (s, 1 H), 8.03 (s, 1 H), 7.16 (brs, 2 H), 5.53 (s, 2 H), 5.50 (s , 2H), 4.28 (t, 2H, J = 5.2 Hz), 4.24-4.21 (m, 1H), 3.93 (d, 2H, J = 7.6 Hz), 3.85 (t, 2H, J = 5.2 Hz), 2.64 to 2.59 (m, 1 H), 2.43 to 2.37 (m, 1 H), 1.10 (s, 18 H)

Comparative Example 7: Preparation of Adefovir Difixyl Benzenesulfonate

Anhydrous adefovir difficile 3.00 g (6.0 mmol) was completely dissolved in 15 ml of acetone, and 1.05 g (6.0 mmol) of benzenesulfonic acid was added and stirred at 20 to 25 ° C. for 2 hours. 25 ml of isopropyl ether was added to the reaction solution, and the temperature of the reaction solution was slowly cooled to 0 to 5 ° C and stirred for 1 hour. The resulting white crystalline solid was filtered and then dried under vacuum at 50 ° C. for 24 hours to give 3.84 g of adefovir dipoxyl benzenesulfonate. The yield was 97.3%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difixyl benzenesulfonate were carried out and the results are shown in FIGS. 17 and 18, respectively.

Melting Point: 128 ~ 132 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.42 (s, 1 H), 8.39 (s, 1 H), 7.58 ~ 7.55 (m, 2H), 7.30 ~ 7.26 (m, 3H) , 5.53 (s, 2H), 5.51 (s, 2H), 4.41-4.39 (m, 2H), 3.94 (d, 2H, J = 7.6 Hz), 3.90-3.88 (m, 2H), 1.08 (s, 18 H)

Comparative Example 8: Preparation of Adefovir Dificil Citrate

3.00 g (6.0 mmol) of anhydrous adefovir difficile and 1.15 g (6.0 mmol) of citric acid were completely dissolved in 60 ml of methanol, and then stirred at 20 to 25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, and then 10 ml of acetone was added to the residue and dissolved. 30 ml of isopropyl ether was added to the reaction solution, and the mixture was stirred at 20 to 25 ° C. for 2 hours. The resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to give 3.53 g of adefovir dipoxyl citrate. The yield was 85.1%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile citrate were carried out and the results are shown in FIGS. 19 and 20, respectively.

Melting Point: 106 ~ 110 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.10 (s, 1H), 8.03 (s, 1H), 7.18 (brs, 2H), 5.53 (s, 2H), 5.50 (s , 2H), 4.29 (t, 2H, J = 5.2 Hz), 3.93 (d, 2H, J = 7.6 Hz), 3.86 (t, 2H, J = 5.2 Hz), 2.67 (dd, 4H , J = 41.6, 15.6 Hz), 1.10 (s, 18 H)

Comparative Example 9: Preparation of Adefovir Difixyl Tartarate

2.50 g (5.0 mmol) of anhydrous adefovir difficile acid and 0.75 g (5.0 mmol) of tartaric acid were completely dissolved in 50 mL of methanol, followed by stirring at 20 to 25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, and 50 ml of acetone was added to the residue, followed by stirring at 20 to 25 ° C for 3 hours. The resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to give 2.65 g of adefovir dipoxyl tartarate. The yield was 81.6%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile tartarate were performed and the results are shown in FIGS. 21 and 22, respectively.

Melting Point: 110 ~ 118 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.10 (s, 1 H), 8.03 (s, 1 H), 7.20 (brs, 2 H), 5.54 (s, 2 H), 5.50 (s , 2 H), 4.30-4.28 (m, 4 H), 3.93 (d, 2 H, J = 7.6 Hz), 3.86 (t, 2 H, J = 5.2 Hz), 1.09 (s, 18 H)

Comparative Example 10 Preparation of Adefovir Dicarboxyl Citrate Hemihydrate

1.00 g (1.4 mmol) of adefovir difficile citrate prepared in Comparative Example 8 were continuously exposed at a temperature of 40 ° C. and a relative humidity of 75% for 24 hours to obtain 1.01 g of adefovir difficile citrate hemihydrate. It was. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile citrate hemihydrate were carried out and the results are shown in FIGS. 23 and 24, respectively.

Melting Point: 116 ~ 118 ℃

Comparative Example 11: Preparation of Adefovir Difixyl Benzenesulfonate Monohydrate

1.00 g (1.5 mmol) of adefovir difixyl benzenesulfonate prepared in Comparative Example 7 were continuously exposed at a temperature of 40 ° C. and 75% relative humidity for 24 hours to give 1.03 g of adefovir difixyl benzenesulfonate monohydrate. Obtained. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difixyl benzenesulfonate monohydrate were carried out and the results are shown in FIGS. 25 and 26, respectively.

Melting Point: 122 ~ 128 ℃

Comparative Example 12: Preparation of Adefovir Difixyl Alpha Ketoglutarate

5.01 g (10.0 mmol) of anhydrous adefovir difficile and 1.46 g (10.0 mmol) of alpha ketoglutaric acid were completely dissolved in 50 mL of isopropanol and stirred at 20-25 ° C. for 2 hours. 300 ml of isopropyl ether was added to the reaction solution, and the mixture was stirred at 20 to 25 ° C. for 24 hours. The resulting white crystalline solid was filtered off, washed with 25 ml of cooled isopropanol and dried under vacuum at 50 ° C. for 24 hours to give 6.30 g of adefovir difficile alpha ketoglutarate. The yield was 97.0%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile alpha ketoglutarate were performed, and the results are shown in FIGS. 27 and 28, respectively.

Melting Point: 88 ~ 92 ℃

¹ H NMR (400 MHz, CDCl ₃ ): δ = 10.40 (brs, 2 H), 8.27 (s, 1 H), 8.17 (s, 1 H), 5.70 to 5.63 (m, 4 H), 4.47 to 4.44 (m, 2H), 3.98-3.96 (m, 2H), 3.89 (d, 2H, J = 7.6 Hz), 2.90 (brs, 2H), 2.77-2.74 (m, 2H), 1.21 ( s, 18 H)

Comparative Example 13: Preparation of Adefovir Dificil Hyfurate

5.01 g (10.0 mmol) of anhydrous adefovir difficile and 1.79 g (10.0 mmol) of hypofuric acid were completely dissolved in 50 mL of isopropanol and stirred at 20-25 ° C. for 1 hour. The reaction solution was concentrated by distillation under reduced pressure, 10 ml of isopropanol and 60 ml of isopropyl ether were added to the residue, and the mixture was stirred at 20 to 25 ° C for 3 hours. Once a solid started to form, it was cooled to 0-5 ° C and further stirred for 2 hours. The resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to give 5.65 g of adefovir dipoxyl hypofurate. Yield 83.0%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile hypofurate were carried out and the results are shown in FIGS. 29 and 30, respectively.

Melting Point: 84 ~ 90 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.81 to 8.78 (m, 1H), 8.09 (s, 1H), 8.03 (s, 1H), 7.84 (d, 2H, J = 7.6 Hz), 7.53 to 7.42 (m, 3 H), 7.15 (s, 2 H), 5.53 (s, 2 H), 5.50 (s, 2 H), 4.28 to 4.27 (m, 2 H), 3.94 to 3.86 (m, 6H), 1.10 (s, 18H)

Comparative Example 14 Preparation of Adefovir Difixyl Crotonate

5.01 g (10.0 mmol) of anhydrous adefovir difficile and 1.79 g (10.0 mmol) of crotonic acid were completely dissolved in 50 mL of isopropanol and stirred at 20-25 ° C. for 1 hour. The reaction solution was evaporated under reduced pressure, concentrated, and then 25 ml of toluene was added to the residue, followed by heating to completely dissolve it. 75 ml of n-hexane was slowly added to the reaction solution, stirred at 20 to 25 ° C. for 2 hours, cooled to 0 to 5 ° C., and further stirred for 2 hours. The resulting white crystalline solid was filtered and dried under vacuum at 50 ° C. for 24 hours to give 5.48 g of adefovir dipoxyl crotonate. The yield was 93.4%. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir difficile crotonate were carried out and the results are shown in FIGS. 31 and 32, respectively.

Melting Point: 72 ~ 74 ℃

¹ H NMR (400 MHz, CDCl ₃ ): δ = 8.29 (s, 1H), 7.97 (s, 1H), 7.08 to 6.99 (m, 1H), 6.76 (brs, 4H), 5.91 to 5.86 (m, 1H), 5.70 to 5.63 (m, 4H), 4.41 (t, 2H, J = 5.2 Hz), 3.95 (t, 4H, J = 4.8 Hz), 3.86 (d, 4 H, J = 7.6 Hz), 1.92-1.90 (m, 3 H), 1.21 (s, 18 H)

Comparative Example 15 Preparation of Adefovir Dificil Hyfurate Dihydrate

1.00 g (1.5 mmol) of adefovir difficile hypofurate prepared in Comparative Example 13 was continuously exposed at a temperature of 40 ° C. and a relative humidity of 75% for 24 hours so that 1.05 g of adefovir difficile hypofurate dihydrate. Obtained. X-ray diffraction analysis and differential scanning calorimetry of the obtained crystalline adefovir dipoxyl hypofurate dihydrate were carried out and the results are shown in FIGS. 33 and 34, respectively.

Melting Point: 64 ~ 68 ℃

Comparative Example 16: Preparation of Adefovir Difixyl Oxalate

1.23 g (10.0 mmol) of oxalic acid dihydrate were completely dissolved in 15 mL of methanol. 5.01 g (10.0 mmol) of anhydrous adefovir difficile was added to another reaction vessel, completely dissolved in 35 ml of methanol, and the solution of the oxalic acid dihydrate was slowly added dropwise for 30 minutes. The reaction solution was stirred at 20 to 25 ° C. for 3 hours, then cooled to 0 to 5 ° C. and further stirred for 2 hours. The resulting crystalline solid was filtered, washed with 10 ml of cooled methanol and dried under vacuum at 50 ° C. for 24 hours to give 5.40 g of adefovir dipoxyl oxalate. The yield was 90.0%.

Melting Point: 164 ~ 166 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.14 (s, 1 H), 8.08 (s, 1 H), 7.47 (brs, 2 H), 5.53 (s, 2 H), 5.50 (s , 2H), 4.30 (t, 2H, J = 5.2 Hz), 3.86 (t, 2H, J = 5.2 Hz), 1.09 (s, 18H)

Comparative Example 17 Preparation of Adefovir Difixyl Methanesulfonate

5.01 g (10.0 mmol) of anhydrous adefovir difficile acid and 0.96 g (10.0 mmol) of methanesulfonic acid were completely dissolved in 50 mL of methanol, followed by stirring at 20 to 25 ° C for 1 hour. The reaction solution was distilled off under reduced pressure, 50 ml of acetone was added to the residue, and the mixture was heated to 50 ° C to completely dissolve it. The reaction solution was slowly cooled to 20-25 ° C. and stirred at the same temperature for 12 hours. The resulting white solid was filtered and dried under vacuum at 50 ° C. for 24 hours to give 5.46 g of adefovir dipoxyl methanesulfonate. The yield was 91.5%.

Melting Point: 140 ~ 142 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.44 (s, 1 H), 8.39 (s, 1 H), 5.52 (s, 2 H), 5.49 (s, 2 H), 4.40 (t , 2H, J = 5.2 Hz), 3.93 (d, 2H, J = 7.6 Hz), 3.89 (t, 2H, J = 5.2 Hz), 2.36 (s, 3H), 1.09 (s, 18H )

Comparative Example 18 Preparation of Adefovir Difixyl Fumarate

5.01 g (10.0 mmol) of anhydrous adefovir difficile acid and 1.16 g (10.0 mmol) of fumaric acid were completely dissolved in 50 ml of methanol, and then stirred at 20 to 25 ° C. for 1 hour. The reaction solution was distilled off under reduced pressure, 50 ml of acetone was added to the residue, and the mixture was heated to 50 ° C to completely dissolve it. The reaction solution was slowly cooled to 20-25 ° C. and stirred at the same temperature for 12 hours. The resulting white solid was filtered and then dried under vacuum at 50 ° C. for 24 hours to give 4.78 g of adefovir dipoxyl fumarate. The yield was 77.4%.

Melting Point: 146 ~ 150 ℃

¹ H NMR (400 MHz, DMSO-d ₆ ): δ = 8.09 (s, 1 H), 8.03 (s, 1 H), 7.15 (brs, 2 H), 6.59 (s, 2 H), 5.53 (s , 2H), 5.50 (s, 2H), 4.28 (t, 2H, J = 5.2 Hz), 3.92 (d, 2H, J = 7.6 Hz), 3.86 (t, 2H, J = 5.2 Hz ), 2.64 to 2.59 (m, 1 H), 2.43 to 2.37 (m, 1 H), 1.10 (s, 18 H)

Test Example 1 Qualitative Confirmation of the Crystalline Adefovir Difficile Acid Addition Structure

1, 3, 5, 7, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 and 33. From the crystalline adefovir difficyl malonate prepared in Examples 1 and 2 and the crystalline adefovir difficile glutarate, respectively, prepared in Comparative Examples 1 to 15, crystalline adefovir difficile L- Mandelate, crystalline adefovir dipoxyl salicylate, crystalline adefovir dipoxyl orotate, crystalline adefovir dipoxyl genitate, crystalline adefovir dipoxyl DL-maleate, crystalline adefovir difixyl benzenesulfonic acid Salt, crystalline adefovir dipoxylate citrate, crystalline adefovir dipoxyl tartarate, crystalline adefovir dipoxylate citrate hemihydrate, crystalline adefovir dipoxyl benzenesulfonate monohydrate, crystalline Defovir difficile alpha ketoglutarate, crystalline adefovir difficile hypofurite, crystalline adefovir difficile crotonate, and crystalline adefovir difficile hypofurlate dihydrate were respectively determined in powder X-ray diffraction analysis. It was confirmed that it had a characteristic crystal form. 1, 3, 5, 7, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 and 33. The characteristic peaks of the powder X-ray diffraction spectra of are shown in Tables 1 to 17, respectively, where '2θ' is a diffraction angle, 'd' is a distance between crystal planes, and 'I / I' ₀ 'means the relative intensity of the peak (peak).

Table 1 X-ray diffraction analysis of adefovir difixyl malonate (crystalline Form I)

2θ d I / I ₀ 2θ d I / I ₀ 8.0745 10.94998 100 22.1117 4.02021 5.86 11.0128 8.03422 6.30 22.6445 3.9268 21.30 12.0606 7.33843 83.98 24.6260 3.61515 15.27 12.4269 7.12298 6.72 26.5880 3.35266 16.93 14.5307 6.09605 2.25 28.1775 3.16705 7.03 16.0622 5.51811 29.00 29.2376 3.05459 6.55 17.1308 5.17622 24.70 29.6318 3.01484 7.92 18.0868 4.90471 3.95 32.2507 2.77575 3.53 19.0792 4.65179 47.85 32.9477 2.71861 15.74 19.5164 4.54856 15.12 34.6059 2.59204 1.69 20.0521 4.42823 9.60 36.8857 2.43691 2.50 20.8816 4.25417 39.25 38.5505 2.33542 3.29 21.6346 4.10776 7.81

Table 2 X-ray diffraction analysis of adefovir difixyl malonate (crystalline Form II)

2θ d I / I ₀ 2θ d I / I ₀ 8.0136 11.03311 8.60 20.5171 4.32892 16.26 9.8430 8.98625 1.74 20.7999 4.27068 14.31 11.0529 8.00514 13.18 21.6876 4.09785 21.26 11.9982 7.37648 8.46 22.0918 4.02378 16.45 12.4280 7.12232 9.03 22.6075 3.93314 24.09 13.4905 6.56367 3.86 22.9891 3.86871 24.44 14.5948 6.06944 16.06 24.7577 3.59621 45.49 16.5236 5.36503 10.17 26.9709 3.30592 9.13 17.1737 5.16340 100.00 27.9278 3.1948 8.35 17.5410 5.05609 22.66 29.6497 3.01306 15.82 18.2552 4.85985 13.78 32.1818 2.78154 3.43 19.0286 4.66405 28.18 32.9341 2.71970 7.80 19.4956 4.55335 40.99 34.7685 2.58030 4.17 19.7859 4.48721 15.99 37.8172 2.37900 2.24

Table 3 X-ray Diffraction Analysis of Adefovir Difixyl Glutarate

2θ d I / I ₀ 2θ d I / I ₀ 5.1701 17.09312 8.61 23.1079 3.8491 0.81 8.511 10.38943 0.36 24.0978 3.69318 0.68 10.3349 8.55964 3.93 26.0478 3.42096 18.46 13.7798 6.42652 1.19 27.6392 3.22749 4.04 14.4262 6.14001 1.10 28.9451 3.08479 1.18 15.5355 5.70397 7.00 31.372 2.85146 8.42 17.195 5.15704 3.67 32.6609 2.74182 1.17 17.7089 5.00853 2.31 35.4248 2.53398 0.29 20.7747 4.27581 100 36.765 2.44463 1.14 22.4494 3.96049 1.43

Table 4 X-ray Diffraction Analysis of Adefovir Difixyl L-Mandelate

2θ d I / I ₀ 2θ d I / I ₀ 6.9222 12.77005 34.22 20.7286 4.28521 34.50 10.3817 8.52112 36.98 23.2373 3.82795 100 10.9665 8.06802 10.64 24.0203 3.70491 70.77 11.8227 7.4856 35.78 25.0998 3.54797 38.44 12.5011 7.08083 18.84 26.308 3.38771 49.67 13.2104 6.7022 4.39 27.1023 3.29019 14.84 14.6466 6.04808 18.13 28.3199 3.15144 51.38 15.2093 5.82557 37.79 29.4681 3.03121 51.95 15.7593 5.62346 33.04 30.2544 2.9542 4.70 15.9444 5.55859 37.09 31.3607 2.85246 11.34 16.2763 5.44598 18.48 32.1462 2.78454 19.25 16.9008 5.24613 91.99 33.0917 2.70711 15.43 17.2389 5.14399 53.69 34.3092 2.61378 8.97 17.519 5.0624 47.76 35.4616 2.53144 5.84 18.42 4.81676 82.68 36.294 2.47527 7.28 19.3141 4.59574 54.91 37.682 2.38723 8.14 20.3122 4.37211 87.70

Table 5 X-ray diffraction analysis of adefovir dipoxyl salicylate

2θ d I / I ₀ 2θ d I / I ₀ 6.9673 12.68754 32.32 22.0084 4.03884 20.40 8.0924 10.92588 5.88 23.3413 3.81113 28.04 8.9773 9.85073 40.73 24.9231 3.57271 10.31 11.5577 7.6566 5.01 25.976 3.43024 8.18 12.7034 6.96852 10.15 26.6078 3.35021 13.44 13.9062 6.36837 28.60 27.8624 3.20215 15.63 15.2726 5.80157 100 28.8948 3.09004 27.06 15.8242 5.60055 21.63 30.6132 2.9204 6.13 17.8781 4.96151 29.67 32.0741 2.79063 3.02 19.241 4.61304 6.48 35.2334 2.54731 1.66 21.0135 4.22775 64.41 37.598 2.39236 2.65

Table 6 X-ray Diffraction Analysis of Adefovir Dificil Orotate

2θ d I / I ₀ 2θ d I / I ₀ 5.8830 15.02315 4.85 17.9847 4.93234 41.67 8.9038 9.93188 29.30 18.6301 4.76290 27.76 10.9992 8.04413 12.99 20.6523 4.30088 10.74 12.3980 7.13948 5.87 23.2501 3.82586 14.46 14.8550 5.96370 14.94 26.6154 3.34927 100 17.1541 5.16924 33.10

Table 7 X-ray Diffraction Analysis of Adefovir Dificent Genitate

2θ d I / I ₀ 2θ d I / I ₀ 7.8918 11.20317 22.91 24.1273 3.68873 81.64 14.0731 6.29323 85.63 26.3857 3.37791 96.03 18.7083 4.74315 100

Table 8 X-ray Diffraction Analysis of Adefovir Difisil DL-Malate

2θ d I / I ₀ 2θ d I / I ₀ 6.6936 13.20566 11.53 21.0564 4.21924 15.97 9.2829 9.52712 7.84 22.4387 3.96234 12.29 10.0089 8.83770 39.06 23.5143 3.78348 25.65 10.7166 8.25561 8.33 24.7631 3.59544 7.05 12.7713 6.93163 41.75 26.3493 3.38249 29.68 13.3599 6.62754 5.54 28.3198 3.15145 19.82 14.6686 6.03907 5.43 29.4430 3.03374 7.39 15.3405 5.77604 7.42 30.5130 2.92975 8.56 16.7061 5.30684 100 32.0054 2.79647 3.54 17.2183 5.15011 37.27 32.8690 2.72493 3.97 18.0539 4.91358 23.74 33.6851 2.66076 5.95 18.4939 4.79768 33.29 35.1027 2.55649 5.29 19.2346 4.61456 17.09 37.4235 2.40311 1.61 19.7042 4.50562 17.33 38.8293 2.31929 2.49 20.6569 4.29993 13.20

Table 9 X-ray Diffraction Analysis of Adefovir Difixyl Benzenesulfonate

2θ d I / I ₀ 2θ d I / I ₀ 6.874 12.85939 1.69 22.7263 3.91286 30.72 8.6425 10.23162 1.01 23.5866 3.77204 13.31 10.1352 8.72785 100 24.2274 3.67372 7.35 11.3136 7.82129 6.68 24.7896 3.59165 5.75 12.4955 7.084 3.56 25.8933 3.44101 17.03 13.4882 6.56479 23.11 27.2783 3.26936 8.77 13.9314 6.35694 4.07 28.6023 3.12097 2.74 15.129 5.85631 6.97 29.1418 3.06441 4.36 15.5911 5.68374 3.72 30.1383 2.96532 4.95 16.4373 5.393 8.06 30.4888 2.93202 5.96 17.6556 5.02354 9.64 32.4184 2.76177 1.94 18.0771 4.90733 14.40 33.4383 2.67984 2.16 19.5787 4.53423 17.30 34.333 2.61202 1.93 20.1787 4.40073 5.63 35.4941 2.5292 2.74 20.6185 4.30785 11.47 36.526 2.46008 3.36 21.1094 4.20876 17.13 37.4261 2.40296 3.30

Table 10 X-ray Diffraction Analysis of Adefovir Dificil Citrate

2θ d I / I ₀ 2θ d I / I ₀ 6.9518 12.71573 26.94 23.9678 3.7129 66.81 8.4195 10.50211 26.63 25.6032 3.47935 39.75 12.7202 6.95938 17.41 26.737 3.33431 26.98 14.4029 6.14985 47.27 28.0005 3.18666 51.98 14.9688 5.9186 42.91 29.3172 3.04647 16.99 15.699 5.64493 74.28 31.0231 2.88274 16.40 17.3572 5.10921 100.00 31.8499 2.80976 9.98 18.982 4.67537 65.34 33.3478 2.6869 7.04 20.7746 4.27583 60.27 35.7424 2.51219 18.70 22.5266 3.94709 53.61

Table 11 X-Ray Diffraction Analysis of Adefovir Difixyl Tartarate

2θ d I / I ₀ 2θ d I / I ₀ 5.7659 15.32805 32.87 19.0613 4.65612 97.59 8.8865 9.95116 54.87 20.7564 4.27953 100 10.3308 8.56301 82.33 21.582 4.11767 65.64 15.5164 5.71094 92.84 26.0801 3.41679 39.22 17.2415 5.14323 59.01 31.5593 2.83496 19.98 18.301 4.8478 84.98 33.9082 2.64377 12.93

Table 12 X-ray diffraction analysis of adefovir dicarboxyl citrate hemihydrate

2θ d I / I ₀ 2θ d I / I ₀ 6.5234 13.54974 21.30 20.2000 4.39615 45.90 8.5116 10.38871 47.56 21.9406 4.05117 24.71 9.8793 8.95327 7.41 23.5475 3.77822 41.79 13.1602 6.72764 8.88 24.4117 3.64639 30.04 14.6083 6.06384 16.72 26.1599 3.40655 16.15 15.1398 5.85217 21.29 27.2199 3.27625 21.38 15.9579 5.55391 35.36 28.1571 3.16930 11.40 16.7360 5.29744 26.08 29.2886 3.04938 13.22 17.7853 4.98718 100 33.0837 2.70774 4.55 18.5256 4.78953 26.78 35.2123 2.54879 11.92

Table 13 X-ray Diffraction Analysis of Adefovir Difixyl Benzenesulfonate Monohydrate

2θ d I / I ₀ 2θ d I / I ₀ 6.9665 12.68892 30.90 22.7564 3.90774 22.32 8.1077 10.90521 31.74 23.3118 3.81588 14.38 10.4439 8.47053 61.23 24.3364 3.65751 26.57 12.9266 6.84869 41.98 24.8015 3.58995 27.69 13.4592 6.57888 17.73 25.9613 3.43215 36.27 13.9012 6.37066 62.81 26.1319 3.41013 27.64 15.7905 5.61244 27.20 27.1602 3.28332 18.07 16.4048 5.40364 100 27.6136 3.23043 12.07 17.3767 5.10353 29.49 28.2227 3.16208 4.77 17.8897 4.95831 17.96 28.7904 3.10101 7.03 18.1043 4.90002 16.20 30.0174 2.97698 10.91 18.8872 4.69863 24.25 30.6727 2.91486 28.40 19.5093 4.55021 74.90 31.5322 2.83734 3.30 20.0020 4.43921 11.35 33.0319 2.71187 5.47 20.9065 4.24915 65.56 34.2006 2.62183 6.28 21.5263 4.12819 9.81 36.5246 2.46017 18.27 21.8232 4.07268 7.71 37.0759 2.42484 4.23 22.4570 3.95915 25.75 39.3223 2.29134 4.34

Table 14 X-ray Diffraction Analysis of Adefovir Difixyl Alpha Ketoglutarate

2θ d I / I ₀ 2θ d I / I ₀ 7.4773 11.82327 100.00 18.8919 4.69749 44.05 8.0776 10.94579 80.56 19.8869 4.46463 59.58 10.3961 8.50936 21.59 21.3636 4.15926 20.00 11.3645 7.78633 5.00 23.1601 3.84053 40.02 12.9283 6.84780 18.92 24.1490 3.68547 75.76 15.4826 5.72333 40.18 25.9995 3.42719 19.17 16.0981 5.50588 51.26 27.3176 3.26476 23.17 16.5588 5.35372 41.96 28.8146 3.09845 12.45 17.8820 4.96044 86.59 31.0262 2.88245 6.69

Table 15 X-Ray Diffraction Analysis of Adefovir Dificil Hyfurate

2θ d I / I ₀ 2θ d I / I ₀ 7.1658 12.33638 48.38 21.8647 4.06505 44.71 7.7324 11.43372 16.35 22.1881 4.00653 10.37 12.1954 7.25763 14.43 22.7274 3.91267 20.88 12.7299 6.9541 25.54 23.3431 3.81084 37.54 12.9729 6.82437 30.34 24.1644 3.68315 7.09 13.7188 6.45495 10.62 24.5169 3.63099 5.17 14.2825 6.20144 6.54 25.3402 3.51486 7.69 15.8888 5.57793 31.59 26.1085 3.41313 7.13 17.3781 5.10311 100 27.9434 3.19305 32.03 17.5619 5.05013 83.25 28.5742 3.12397 11.33 17.9864 4.93187 8.82 28.9090 3.08855 12.19 19.3717 4.58221 6.22 31.3412 2.85420 5.91 19.9065 4.46029 5.59 32.7923 2.73114 10.32 20.9752 4.23538 86.38 34.6319 2.59016 7.85 21.4183 4.14876 54.67 36.3820 2.46948 2.54 21.6334 4.10800 69.29 37.0556 2.42613 2.06

Table 16 X-ray Diffraction Analysis of Adefovir Difixyl Crotonate

2θ d I / I ₀ 2θ d I / I ₀ 5.0211 17.60004 100 21.9033 4.05798 15.21 6.1699 14.32521 15.55 22.9225 3.8798 12.88 7.9157 11.16931 25.15 23.4646 3.79139 19.92 9.9614 8.87966 45.00 24.2045 3.67714 19.70 11.6678 7.58464 16.17 25.6869 3.46819 33.17 12.2498 7.22555 6.02 26.5798 3.35368 11.40 13.2299 6.69237 16.80 28.0079 3.18583 10.21 14.92 5.93786 10.63 28.8148 3.09843 6.52 15.7877 5.61344 40.99 30.5959 2.922 9.00 16.1214 5.49798 31.21 31.8202 2.81232 3.44 16.7291 5.2996 3.70 33.0811 2.70795 8.29 17.3723 5.10481 14.25 34.1781 2.62351 3.05 18.1156 4.897 61.48 35.8064 2.50785 5.27 19.1252 4.6407 71.40 37.0365 2.42734 3.68 20.1026 4.41722 13.51 37.6363 2.39002 4.16 20.8489 4.26076 28.94 38.3221 2.34881 3.23 21.2569 4.17989 28.23

Table 17 X-ray Diffraction Analysis of Adefovir Difixyl Hipfurate Dihydrate

2θ d I / I ₀ 2θ d I / I ₀ 8.5550 10.33602 43.68 21.8628 4.06540 100 8.7910 10.05913 39.65 22.2329 3.99857 20.77 9.5402 9.27076 23.72 23.4065 3.80066 36.63 11.1927 7.90546 35.35 23.7526 3.74606 13.12 12.3386 7.17372 10.71 24.4388 3.64242 73.50 12.9812 6.82001 20.42 25.2864 3.52221 16.21 14.4732 6.12013 15.51 25.8144 3.45135 16.89 15.0840 5.87367 6.11 28.0079 3.18584 63.66 16.3643 5.41689 77.23 28.8912 3.09041 18.46 16.8867 5.25050 18.22 30.2244 2.95707 13.43 17.6474 5.02584 29.17 30.9022 2.89374 16.43 18.3651 4.83103 26.85 31.6748 2.82489 5.89 18.8232 4.71446 18.98 32.8065 2.72999 29.95 19.7478 4.49577 61.80 34.4275 2.60507 7.16 21.0005 4.23035 47.26 35.8029 2.50809 4.58 21.3333 4.16509 59.98 37.0164 2.42861 6.10

Test Example 2: Stability Test for Moisture and Heat

Moisture and heat stability of the active ingredient used in the pharmaceutical composition is an important factor not only in the manufacturing process of the composition but also in the long term storage of the composition, so that anhydrous adefovir difficile having a structure of Form 1 (see US Patent No. 6645340 And Korean Patent Registration No. 618663) and the crystalline adefovir dipoxyl malonate prepared in Examples 1 and 2 and the crystalline adefovir difixyl glutarate prepared in Comparative Examples 1 to 18, respectively. , Crystalline adefovir dipoxyl L-mandelate, crystalline adefovir dipoxyl salicylate, crystalline adefovir dipoxyl orotate, crystalline adefovir dipoxyl gentiate, crystalline adefovir dipoxyl DL-maleate Crystalline Adefovir Difixyl Benzenesulfonate, Crystalline Adefovir Difixyl Citrate, Sex Adefovir Dicarboxyl Tartarate, Crystalline Adefovir Dicarboxyl Citrate Hemihydrate, Crystalline Adefovir Dicarboxyl Benzenesulfonate Monohydrate, Crystalline Adefovir Difibox Alphaketoglutarate, Crystalline Adefovir Dicoat Silhifurate, Crystalline Adefovir Difficile Crotonate, Crystalline Adefovir Difficile Hypurate Dihydrate, Crystalline Adefovir Difixyl Oxalate (see Chinese Patent Publication No. 1528766), Crystalline adefovir diecoat Moisture of the real methanesulfonic acid salt (cf. U.S. Pat. The stability to heat was measured. Specifically, 0, 3, 7, 14 and 28 days of each of the above-described adefovir difficile acid and their acid addition salts were stored in a sealed state under severe conditions of 40 ° C. and 75% relative humidity. Residual ratios of initial active ingredient values were analyzed by high performance liquid chromatography (HPLC) for the samples afterwards. The results are shown in Table 18 below.

Table 18 Stability Test for Moisture and Heat

salt Early 3 days 7 days 14 days 28 days Adefovir Daifi Room 100.0 100.0 99.8 98.8 96.7 Adefovir dipoxyl malonate (crystalline Form I) 100.0 99.9 99.9 99.8 99.3 Adefovir Difixyl Malonate (crystalline Form II) 100.0 99.9 99.9 99.6 99.0 Adefovir Difisil Glutarate 100.0 95.2 89.3 80.4 69.2 Adefovir Difisil L-Mandelate 100.0 98.9 97.4 96.4 94.5 Adefovir Difisil Salicylate 100.0 98.8 97.2 94.8 90.1 Adefovir Difisil Orotate 100.0 97.8 94.5 88.4 65.7 Adefovir Dificent Gentiate 100.0 98.6 94.9 90.5 73.5 Adefovir Difisil DL-Malate 100.0 98.7 97.6 95.4 86.6 Adefovir Difixyl Benzenesulfonate 100.0 98.9 98.3 90.7 67.5 Adefovir difficile citrate 100.0 97.7 94.3 88.9 83.6 Adefovir difficile tartarate 100.0 98.8 95.6 87.1 76.6 Adefovir dipoxylate citrate hemihydrate 100.0 99.4 97.0 94.3 90.5 Adefovir Difixyl Benzenesulfonate Monohydrate 100.0 99.0 98.7 92.8 75.8 Adefovir difficile alpha ketoglutarate 100.0 99.4 97.4 84.5 30.2 Adefovir Difisil Hypurate 100.0 99.2 99.6 95.7 78.9 Adefovir difficile crotonate 100.0 99.0 98.2 94.6 86.7 Adefovir Dififil Hyfurate Dihydrate 100.0 100.0 99.2 99.1 96.6 Adefovir Difixo Oxalate 100.0 99.8 99.3 98.9 97.0 Adefovir Difisil Methanesulfonate 100.0 98.0 93.3 87.6 70.1 Adefovir Difisil Fumarate 100.0 98.8 95.8 86.8 77.1

As shown in Table 18 above, the adefovir dipoxyl malonate of Form I and Form II was significantly superior to the adefovir dipivox of Form 1, as well as other acid addition salts of adefovir dipivoxil in a 28 day harsh condition experiment. Showed stability. From these results, it was confirmed that the crystalline adefovir difficile malonate of the present invention has a chemical stability that can be usefully used as a pharmaceutical composition for the treatment of HIV and hepatitis B virus infection.

Test Example 3: Nonhygroscopic Test

Crystalline adefovir dipoxyl malonate prepared in Examples 1 and 2, and crystalline adefovir dipoxyl glutarate prepared in Comparative Examples 1 to 18, respectively, and crystalline adefovir difficile L-mandel Acid salts, crystalline adefovir dipoxyl salicylate, crystalline adefovir dipoxyl orotate, crystalline adefovir dipoxyl genitate, crystalline adefovir dipoxyl DL-maleate, crystalline adefovir difixyl benzenesulfonate , Crystalline adefovir dipoxylate citrate, crystalline adefovir dipoxylate tartarate, crystalline adefovir dipoxylate citrate hemihydrate, crystalline adefovir diefixyl benzenesulfonate monohydrate, crystalline adefovir difibox alphaketo Glutarate, Crystalline Adefovir Difisil Hypurate, Crystalline Adefovy Hygroscopicity for Dipicyl Crotonate, Crystalline Adefovir Diphyxyl Hyfurate Dihydrate, Crystalline Adefovir Dipicyl Oxalate, Crystalline Adefovir Dipicyl Methanesulfonate and Crystalline Adefovir Dificil Fumarate It was compared with the hygroscopicity of the anhydrous adefovir difficile chamber having a structure of 1. After each compound was continuously exposed for 2 hours, 8 hours, 24 hours and 3 days at a temperature of 40 ° C. and a relative humidity of 75%, the moisture content was measured with a Karl-Fisher moisture meter. . Table 19 shows the measured values of the water content of the active ingredient in terms of moisture content (% by weight).

Table 19 Nonhygroscopic Test

salt Early 2 hours 8 hours 24 hours 3 days Adefovir Daifi Room 0.25 2.40 5.49 7.55 7.57 Adefovir dipoxyl malonate (crystalline Form I) 0.22 0.21 0.24 0.26 0.24 Adefovir Difixyl Malonate (crystalline Form II) 0.26 0.26 0.30 0.33 0.36 Adefovir Difisil Glutarate 0.19 0.26 0.27 0.26 0.26 Adefovir Difisil L-Mandelate 0.14 0.15 0.16 0.16 0.15 Adefovir Difisil Salicylate 0.15 0.16 0.20 0.25 0.26 Adefovir Difisil Orotate 1.02 1.52 1.51 1.81 1.92 Adefovir Dificent Gentiate 0.77 0.76 0.77 0.81 0.87 Adefovir Difisil DL-Malate 0.17 0.19 0.29 0.32 0.37 Adefovir Difixyl Benzenesulfonate 0.22 3.30 3.31 3.31 3.35 Adefovir difficile citrate 0.31 0.51 1.74 1.94 2.01 Adefovir difficile tartarate 0.34 0.52 0.53 0.53 0.56 Adefovir dipoxylate citrate hemihydrate 1.67 1.95 1.95 2.03 2.25 Adefovir Difixyl Benzenesulfonate Monohydrate 3.31 3.31 3.30 3.32 3.35 Adefovir difficile alpha ketoglutarate 0.48 0.48 0.49 0.50 0.56 Adefovir Difisil Hypurate 0.34 0.36 0.38 2.51 5.69 Adefovir difficile crotonate 0.35 0.47 0.66 1.31 2.23 Adefovir Dififil Hyfurate Dihydrate 5.69 5.68 5.69 5.72 5.74 Adefovir Difixo Oxalate 0.15 0.24 0.26 0.28 0.29 Adefovir Difisil Methanesulfonate 0.20 0.21 0.25 0.28 0.37 Adefovir Difisil Fumarate 0.22 0.29 0.32 0.33 0.38

As shown in Table 19, the adefovir difficile malonate of Form I and Form II showed significantly better non-hygroscopicity than the anhydrous adefovir Dificil having the structure of Form 1. From these results, it was confirmed that the crystalline adefovir difficyl malonate according to the present invention is relatively stable to moisture and thus can be usefully used as an active ingredient of the pharmaceutical composition.

Test Example 4: Solubility test in pH range in vivo

The solubility of the active ingredient of the pharmaceutical composition in aqueous solution will affect the dissolution rate of the composition, which will greatly affect the bioavailability of the drug. Crystalline adefovir dipoxyl malonate prepared in Examples 1 and 2, and crystalline adefovir dipoxyl glutarate prepared in Comparative Examples 1 to 18, respectively, and crystalline adefovir difficile L-mandel Acid salts, crystalline adefovir dipoxyl salicylate, crystalline adefovir dipoxyl orotate, crystalline adefovir dipoxyl genitate, crystalline adefovir dipoxyl DL-maleate, crystalline adefovir difixyl benzenesulfonate , Crystalline adefovir dipoxylate citrate, crystalline adefovir dipoxylate tartarate, crystalline adefovir dipoxylate citrate hemihydrate, crystalline adefovir diefixyl benzenesulfonate monohydrate, crystalline adefovir difibox alphaketo Glutarate, Crystalline Adefovir Difisil Hypurate, Crystalline Adefovy The solubility of dipoxyl crotonate, crystalline adefovir dipoxyl dipuric acid dihydrate, crystalline adefovir dipoxyl oxalate, crystalline adefovir dipoxyl methanesulfonate and crystalline adefovir dipoxyl fumarate was determined in crystalline form 1. The solubility of the anhydrous adefovir difficile acid having a structure was compared. Specifically, the solubility experiment of the active ingredient was conducted in the pH range in vivo required for bioabsorption, i.e., pH (1.2) in the stomach, pH (5.2) in the intestine and pH (7.4) close to the blood pH (7.4). After dissolving each of the adefovir difficile acid addition salts and adefovir dificil to reach a saturation state, the solution was analyzed by high performance liquid chromatography (HPLC) and dissolved on the basis of the adefovir difficile base (free base). The amount was measured. The results are shown in Table 20 below.

Table 20 Solubility Test

salt Deionized water
(mg / ml) pH 1.2
(mg / ml) pH 5.2
(mg / ml) pH 7.4
(mg / ml) Adefovir Daifi Room 0.24 32.10 0.24 0.19 Adefovir dipoxyl malonate (crystalline Form I) 1.94 36.28 0.54 0.29 Adefovir Difixyl Malonate (crystalline Form II) 1.72 41.66 0.37 0.32 Adefovir Difisil Glutarate 0.66 17.70 0.25 0.18 Adefovir Difisil L-Mandelate 1.05 17.17 0.35 0.21 Adefovir Difisil Salicylate 0.81 8.13 0.31 0.20 Adefovir Difisil Orotate 1.60 2.91 0.44 0.18 Adefovir Dificent Gentiate 0.90 7.39 0.27 0.19 Adefovir Difisil DL-Malate 0.97 34.0 0.28 0.20 Adefovir Difixyl Benzenesulfonate 3.53 0.16 0.36 0.19 Adefovir difficile citrate 0.99 39.78 0.34 0.20 Adefovir difficile tartarate 0.29 38.20 0.64 0.20 Adefovir dipoxylate citrate hemihydrate 2.39 108.84 0.34 0.24 Adefovir Difixyl Benzenesulfonate Monohydrate 7.49 7.15 0.23 0.20 Adefovir difficile alpha ketoglutarate 10.55 69.21 1.05 0.34 Adefovir Difisil Hypurate 1.43 19.79 0.40 0.34 Adefovir difficile crotonate 0.64 33.55 0.36 0.38 Adefovir Dififil Hyfurate Dihydrate 0.65 17.17 0.40 0.26 Adefovir Difixo Oxalate 4.49 5.94 0.80 0.19 Adefovir Difisil Methanesulfonate 4.60 429.15 0.35 0.19 Adefovir Difisil Fumarate 9.27 10.02 0.35 0.20

As shown in Table 20, the adefovir difixyl malonate of Form I and Form II showed better solubility than the anhydrous adefovir difixyl having a structure of Form 1 in all pH ranges.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a diagram showing X-ray diffraction spectroscopy of the crystalline adefovir difixyl malonate (crystalline Form I) obtained in Example 1.

Figure 2 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difficyl malonate (crystalline Form I) obtained in Example 1.

3 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir difixyl malonate (crystalline Form II) obtained in Example 2. FIG.

Figure 4 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difixyl malonate (crystalline Form II) obtained in Example 2.

FIG. 5 is a diagram showing an X-ray diffraction spectrum diagram of the crystalline adefovir difixyl glutarate obtained in Comparative Example 1. FIG.

Figure 6 is a diagram showing the differential scanning calorimetry of the crystalline adefovir dipoxyl glutarate obtained in Comparative Example 1.

FIG. 7 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir difficile L-mandelate obtained in Comparative Example 2.

8 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difixyl L-mandelate obtained in Comparative Example 2. FIG.

9 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir dipoxyl salicylate obtained in Comparative Example 3. FIG.

10 is a diagram showing the differential scanning calorimetry of the crystalline adefovir dipoxyl salicylate obtained in Comparative Example 3.

FIG. 11 is a diagram showing an X-ray diffraction spectrum diagram of the crystalline adefovir dipoxyl orotate obtained in Comparative Example 4. FIG.

12 is a diagram showing the differential scanning calorimetry of the crystalline adefovir dipoxyl orotate obtained in Comparative Example 4. FIG.

Fig. 13 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir dipoxyl genitate obtained in Comparative Example 5.

14 is a diagram showing the differential scanning calorimetry of the crystalline adefovir dipoxyl gentiate obtained in Comparative Example 5. FIG.

15 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir dipoxyl DL-maleate obtained in Comparative Example 6. FIG.

FIG. 16 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difficile DL-maleate obtained in Comparative Example 6. FIG.

Fig. 17 shows the X-ray diffraction spectroscopy of the crystalline adefovir difixyl benzenesulfonate obtained in Comparative Example 7.

FIG. 18 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difixyl benzenesulfonate obtained in Comparative Example 7.

19 is a diagram showing an X-ray diffraction spectroscopy of the crystalline adefovir dipoxyl citrate obtained in Comparative Example 8. FIG.

20 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difficile citrate obtained in Comparative Example 8.

Fig. 21 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir dipoxyl tartarate obtained in Comparative Example 9.

Fig. 22 is a diagram showing the differential scanning calorimetry of the crystalline adefovir dipoxyl tartarate obtained in Comparative Example 9.

FIG. 23 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir dipoxyl citrate hemihydrate obtained in Comparative Example 10.

FIG. 24 is a diagram showing the differential scanning calorimetry of the crystalline adefovir dicarboxyl citrate hemihydrate obtained in Comparative Example 10. FIG.

25 is a diagram showing an X-ray diffraction spectrum diagram of the crystalline adefovir difixyl benzenesulfonate monohydrate obtained in Comparative Example 11. FIG.

Fig. 26 shows the differential scanning calorimetry of the crystalline adefovir difixyl benzenesulfonate monohydrate obtained in Comparative Example 11.

FIG. 27 is a diagram showing an X-ray diffraction spectrum of crystalline adefovir dipoxyl alpha ketoglutarate obtained in Comparative Example 12.

28 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difficile alpha ketoglutarate obtained in Comparative Example 12. FIG.

FIG. 29 is a diagram showing an X-ray diffraction spectrum diagram of the crystalline adefovir dipoxyl hypofuric acid salt obtained in Comparative Example 13. FIG.

30 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difficile hypofuric acid salt obtained in Comparative Example 13. FIG.

Fig. 31 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir dipoxyl crotonate obtained in Comparative Example 14.

32 is a diagram showing the differential scanning calorimetry of the crystalline adefovir difficile crotonate obtained in Comparative Example 14. FIG.

FIG. 33 is a diagram showing an X-ray diffraction spectrogram of the crystalline adefovir dipoxyl hypofurate dihydrate obtained in Comparative Example 15.

Fig. 34 is a diagram showing the differential scanning calorimetry of the crystalline adefovir dipoxyl hypofurate dihydrate obtained in Comparative Example 15.

Claims (6)

delete delete Adefovir difixyl malonate represented by the following general formula (I) of crystallinity, wherein in X-ray diffraction analysis, I / I ₀ (I: intensity of peak at each diffraction angle, I ₀ : intensity of largest peak) The diffraction angle (2θ) of 10% or more is 8.1 ± 0.2, 12.1 ± 0.2, 16.1 ± 0.2, 17.1 ± 0.2, 19.1 ± 0.2, 19.5 ± 0.2, 20.9 ± 0.2, 22.6 ± 0.2, 24.6 ± 0.2, 26.6 ± 0.2 Adefovir dipoxyl malonate, characterized in that 32.9 ± 0.2:

Adefovir difixyl malonate represented by the following general formula (I) of crystallinity, wherein in X-ray diffraction analysis, I / I ₀ (I: intensity of peak at each diffraction angle, I ₀ : intensity of largest peak) The diffraction angle (2θ) of 10% or more is 11.1 ± 0.2, 14.6 ± 0.2, 16.5 ± 0.2, 17.2 ± 0.2, 17.5 ± 0.2, 18.3 ± 0.2, 19.0 ± 0.2, 19.5 ± 0.2, 19.8 ± 0.2, 20.5 ± 0.2 Adefovir dipoxyl malonate, characterized in that 20.8 ± 0.2, 21.7 ± 0.2, 22.1 ± 0.2, 22.6 ± 0.2, 23.0 ± 0.2, 24.8 ± 0.2, 29.6 ± 0.2:

A pharmaceutical composition for the treatment of HIV or hepatitis B virus infection comprising the adefovir difficile malonate according to claim 3 or 4 together with a pharmaceutically acceptable carrier. A pharmaceutical composition according to claim 5 for the treatment of chronic hepatitis B virus infection.

Images