WO2015163724A1

WO2015163724A1 - Solid form antiviral agent and preparation method therefor

Info

Publication number: WO2015163724A1
Application number: PCT/KR2015/004101
Authority: WO
Inventors: 지준홍; 유형철; 김재선; 엄기안; 이정범; 이보람
Original assignee: 지준홍; 제이투에이치바이오텍 (주); 주식회사 휴온스
Priority date: 2014-04-25
Filing date: 2015-04-24
Publication date: 2015-10-29
Also published as: KR101548724B1

Abstract

The present invention relates to a novel crystal form of Tenofovir disoproxil free base. The amount of related substances generated according to an elapsed time can be minimized in the Tenofovir disoproxil free base, according to the present invention, compared with Tenofovir disoproxil fumarate or a conventional Tenofovir disoproxil free base, and thus the stability of a preparation can be increased by lowering the amount of generated impurities in a product storage process. In addition, the novel crystal form of Tenofovir disoproxil free base of the present invention has improved hygroscopicity, excellent solubility according to pH, and excellent physical and chemical properties, and thus can be used as a useful active ingredient of a pharmaceutical composition.

Description

【Specification】

[Name of invention]

Antiviral agent in solid form and preparation method thereof

This patent application is filed with the Korean Patent Application No. 10-2014—0050011 filed with the Korean Patent Office on April 25, 2014 and the Korean Patent Application No. 10-2014-0151114 filed with the Korean Patent Office on November 03, 2014. Priority is claimed, the disclosures of which are incorporated herein by reference.

The present invention relates to a novel crystalline form of tenofovirisoproxyl free base and a process for preparing the same.

[Background technology]

Tenofovirdisopoxyfumarate (trade name Viread, hereinafter TenenofovirDisoproxilFumarate) has the chemical name 9- [2— (R) ᅳ

[[Bis [[(isopropoxycarbonyl) oxy] methoxy] phosphinoyl] methoxy] propyl] adenine fumaric acid (1: 1) (CAS Registry number 201238-50-9). Tenofodisodixoxyfumarate has a molecular weight of 635.52 and is represented by the following structural formula ().

Tenofovirodisulfoxyfumarate is a nucleotide analog reverse transcriptase and HBV polymerase inhibitor, as determined by Giliard Science. It is developed and marketed under the trade name Vi read, and is a useful drug for the treatment of HIV-1 infection and chronic hepatitis B.

Tenofovirisofufosilfumarate is an crystalline solid and has an endothermic peak of about 118 ^° as measured in DSC (Di f ierental scanning calorimetry) as described in the prior publications WO099 / 05150 and EP998480. Tenofovirisofufocilfumarate is known to have a variety of crystal polymorphs, and the transition of the crystal form may occur due to formulation preparation and change over time. On the other hand, it is known that tenofovirisoproxyl has a problem in stability due to the influence of moisture. In the published literature (Pharmaceutical Research, 18: 234-237 (2001); Pharmaceutical Research, 17: 1098-1103 (2000)), tenofovirdisoproxil is hydrolyzed in the presence of water to generate formaldehyde, It has been reported that the amine groups of the formed formaldehyde and tenofovirodisoxyl cause a condensation reaction to generate Tenenofovirdisoproxil Dimer as an impurity. Thus, preparations of tenofovirisofufolmulfate are essential to strict control and to maintain dry conditions during packaging and storage to prevent moisture absorption.

According to the International Pharmacopoeia of the World Health Organization (WHO), the structure of several flexible substances is known in the case of tenofobicisopropoxyfumarate, and optical isomers, hydrolyzates, hydrolyzed and alkyl substituted substances and dimers ( Dimer) and 13 hydrolysates or all hydrolyzed ones and combinations of these flexible materials have 13 flexible materials to be managed and stored as 2-8 degrees Celsius in a shaded and sealed container as storage conditions. It is supposed to be kept in a condition. On the other hand, 9-prop-1-enyl-9H-purinium 6-amine (9- (prop-l-enyl) — 9H-purin-6-amine), which is designated as analog K, is suspected to be genotoxic. It is regulated to control to 5 ppm or less in the drug substance.

Previous publications US2004 / 0018150, US6,465,649, US5, 935, 946, and US5,977,089 describe the synthesis of tenofovirisoproxyl and salts thereof from tenofovirodisoxyl freebases, which are in the form of mucus oils. Tenofovirisophthoxy fumarate can be prepared or tenofovirisophthoxylate A method of producing tenofovirisofufosilate is used without isolation.

When tenofovirodisotoxyl is reacted in the form of an oil to obtain a salt or to proceed with a salt formation reaction without obtaining a salt (i so l at i on), various problems may occur. Compared with solid form, tenofovirisoproxil in oil form has problems of stability, difficulty in controlling equivalent weight according to salts and exact molar ratio, and difficulty in strictly managing below-standard flexible compounds of acid addition salts. There is this. For example, excess organic acid may be precipitated and mixed during salt formation. It is likely that the drug does not meet the requirements for leaded substances in pharmaceuticals that require strict control standards.

United States Patent Application Publication No. 2011/0112292 discloses a method for preparing a crystalline tenofovirosophyxyl free base to solve this problem; It presents a novel crystal form, and a method of producing with a melting point in C to 66 ^'C. However, when it has such a low melting point, it is difficult to use as an active ingredient of pharmaceuticals because of its low thermal stability, and the present invention also has the usefulness of crystalline tenofovirisophoxyl as an intermediate for preparing tenofovirisophthoxyfumarate To emphasize.

Published Patent Publication No. 2012-0095874 discloses a novel salt of tenofovirisoproxyl and a method for preparing the same, and also discloses a method of obtaining a free base from tenofovirdisofoxyl fumarate. In the case of the free base disclosed in the present invention, ethyl acetate is added to the fumarate, and the resultant is solidified by stirring with sodium bicarbonate, followed by drying and concentration. The free base disclosed in this invention was used as a raw material for preparing aspartate, which is a crystalline acid addition salt, instead of being used as a drug substance.

As a result of analyzing and preparing a glass base according to the method disclosed in Korean Laid-Open Patent Publication No. 2012-0095874, the present inventor's stone is a sticky solid having a melting point formed at 94-97 ^° C., resulting in an uneven particle size distribution. When tablet tableting, adhesion becomes a problem, stability and hygroscopicity are poor, and purity products required for the drug substance are not obtained. It was required, and the amount of residual solvent after drying did not conform to the ICH (International Standard) standard, and thus could not be used as a pharmaceutical raw material.

Therefore, the present inventors have a thermodynamically stable and hygroscopicity, and thus can be stored for a long time at room temperature, and they are not sticky and have excellent solubility. I want to.

Although crystalline acid addition salts of tenofovirisoproxyl have been disclosed in various crystal polymorphisms in a number of prior inventions, they are more stable than existing crystalline acid addition salts, and the specifications and pharmacologically required physical and chemical properties required by the drug substance. The crystalline free bases which stratify have not been reported in the prior invention or literature.

. When crystalline free base is used instead of crystalline acid addition salt as a medical ingredient

One step can be reduced in the production process of GMP raw material, which is also industrially meaningful. The manufacturing method disclosed in the present invention is very efficient and economical because the production yield is very high and high purity does not require additional purification. Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

[Content of invention]

[Problem to solve]

The present inventors have attempted to obtain a novel crystalline free base of tenofovirisopropyl having excellent physicochemical properties and stability while overcoming the drawbacks of the conventional tenofovirisophyxyl fumarate.

As a result, the present inventors have developed a method and conditions capable of preparing a new crystalline form of tenofovirisoproxyl free base having the above-mentioned advantages, and thus the tenofovirisoproxyl freebase prepared is thermodynamically It was found to be stable and, by itself, of high purity suitable for use as an active ingredient in pharmaceuticals. Accordingly, it is an object of the present invention to provide a novel crystalline tenofovirodisoxyl free base.

It is another object of the present invention to provide a method for preparing a crystalline tenofovirisoxopropyl free base.

Another object of the present invention is to provide an antiviral pharmaceutical composition comprising the above-mentioned crystalline tenofovirodisoxyl free base of the present invention. Other objects and advantages of the present invention will become apparent from the following detailed claims and drawings.

[Measures of problem]

According to one aspect of the present invention, the present invention has a positive peak diffraction angle (2Θ ± 0.Γ) in an X-ray diffraction (PXRD) analysis of 7.4 ± 0.2 ^° , 8.1 ^° 0.2 ^° , 10.8 ± 0.2 ^° ,

13 ± 0.2 ^° , 14.4 ± 0.2 ^° , 14.8 ± 0.2 ^° , 16.2 士 0.2 ^° , 16.7 ± 0.2 ^° , 17.6 ± 0.2 ^° 18.04 士 0.2 ^° , 19.19 ± 0.2 ^° , 19.4 土 0.2 ^° , 20.9 士 0.2 ^° , 22.9 2 ^° 24.4 ^° 0.2 ^° , 26.7 ^° 0.2 ° and 28.6 ° 0.2 ^° , and have a differential scanning calorimetry (DSC) characteristic of FIG. 2 and a melting point in the range of 107 ± 3 ^° C. It provides a crystalline tenofovirisoofyl free base:

Formula 2

The present inventors have attempted to obtain a novel crystalline free base of tenofovirisophoxyl having excellent purity, excellent physicochemical properties, and stability while overcoming the disadvantages of the conventional tenofovirdisopoxylfumarate.

As a result, the present inventors have developed a method and conditions capable of preparing a new crystalline form of tenofovirisoproxyl free base having the above-mentioned advantages, and thus the tenofovirisoproxyl free base is thermodynamically stable, It was found to be suitable for use as an active ingredient of medicines by themselves.

The crystalline tenofovirodisoficyl freebase of the present invention represented by the formula (2) exhibits an X-ray diffraction pattern different from other known freebases. According to one embodiment of the present invention, the crystalline tenofovirodisoxyl free base of the present invention has the X-ray diffraction pattern shown in FIG. More specifically, the crystalline tenofovirosophyxyl free base of the present invention has a differential scanning calorimetry (DSC) characteristic of FIG. 2 and a melting point of 107 Pa 3 ^° C. The crystalline tenopovirdisopoxyl free base of the present invention is a conventional free base (e.g., US Patent Publication No. 2011/0112292 and Korean Patent Application Publication No. 2012-0095874) and is currently used as a raw material for pharmaceuticals Compared to povidisopoxyl fumarate, it has excellent stability, meets the specifications of flexible materials and residual solvents, and is a crystalline solid without hygroscopicity and tackiness. According to another aspect of the present invention, the present invention provides a process for the preparation of crystalline tenofovirisoopoxyl free base comprising the following steps:

(a) dissolving or suspending tenofovirodisoxyl acid addition salt in a mixed solvent of methylene chloride or methylene chloride and methanol, adjusting the pH by adding an aqueous solution of sodium carbonate or sodium hydrogen carbonate, and then separating the organic layer;

(b) adding magnesium sulfate (MgSO ₄ ) or sodium sulfate (Na ₂ SO ₄ ) as an absorbent and stirring the resultant of step (a); (c) filtering the resultant of step (b) and concentrating and drying under reduced pressure;

(d) of step (c). Ethanol, isopropanol; Or adding a co-solvent of ethanol and isopropanol, and obtaining crystalline tenofovirodisopropyl free base through vigorous stirring and selective seeding. The manufacturing method of the present invention will be described in detail for each step as follows:

Step (a): Tenofovirosophyxyl acid addition salt treatment

In the present invention, the step of dissolving or suspending tenofovirisoxolic acid addition salt in a mixed solvent of methylene chloride or methylene chloride and methanol, and adjusting the pH by adding an aqueous solution of sodium carbonate or sodium hydrogen carbonate to separate the organic layer by extraction Include.

On the other hand, the treatment of the aqueous sodium carbonate or sodium bicarbonate solution and the mixed solvent of methylene chloride or methylene chloride and methanol added to tenofovirisoxolate acid addition salt is not restricted in that order.

Tenofovirosophyxyl acid addition salts used in the present invention include various acid addition salts of tenofovirisoproxyl, for example, tenofovirisoxofumarate can be used.

PH adjustment by treatment of aqueous sodium carbonate or sodium bicarbonate solution means to create a condition of pH 9-10. Steps (b) — (c): Obtaining Crystalline Tenofovirodisopropyl Free Base by Recrystallization

In step (b), magnesium sulfate (MgS0 ₄ ) or sodium sulfate (Na ₂ S0 ₄ ) is added to the moisture absorbent and stirred to remove water. In step (c), the resultant is preferably filtered at 5 ^° C., concentrated under reduced pressure and dried at 50 ^° C. or lower. Step (d): == r of crystalline tenofovirisoxosiloxane free base by recrystallization

To the product of step (c) or to the starting material of step (a) (ii), ethanol or isopropanol and a co-solvent of the two solvents are added and vigorous stirring and optional seeding as necessary. To obtain a crystalline tenofovirisoopoxyl free base.

Ethanol or isoprapan and the mixing ratio of the two solvents can be prepared in various ways, and the amount of the solvent used is 10-50 ml with respect to 1 g of tenofovirdisopoxyl free base.

It is advantageous to vigorously stir the tenofovirisoxoyl free base and solvent for crystallization, the stirring time being 1-8 hours, more specifically 1-4 hours.

Upon stirring, the tenofovirodixosiloxane crystalline free base according to the present invention can be selectively seeded to speed up the precipitation time of the crystal. Seeding is not essential but is effective when no crystalline solid is precipitated after several hours of stirring. According to another aspect of the invention, the invention provides a process for the preparation of crystalline tenofovirisotropoxyl bases comprising the following steps:

(a) ethanol and isopropanol in the slime free, amorphous free or crystalline free base of tenofovirisoxo; Or adding ethane and a co-sol vent of isopropanol;

(b) filtering the product of step (a);

(c) obtaining the crystalline tenofovirosophyxyl free base through selective seeding of the resultant of step (b). . The manufacturing method of the present invention will be described in detail for each step as follows:

Steps (a)-(b): Treatment of tenofovirisoproxyl freebases (mucous oils or amorphous and crystalline solids) In the preparation method, instead of tenofovirisoxolate addition salts, mucous free bases or amorphous and other forms of crystalline freebases (for example, Korean Laid-Open Patent Publication No. 2012-0095874) are prepared as starting materials. (preparat ion).

A tenofovirisoproxyl free base using a tenofovirisoxoyl free base as a starting material to obtain a crystalline tenofovirisoopyl freebase, which is a slime, amorphous or crystalline tenofovirosophyloxyl free base different from the crystalline form of the present invention. Is the starting material.

Ethane isopropanol in the crystalline tenofovirodixopropyl free base of the above-mentioned mucoid oil, amorphous form or other forms of the present invention; Or co-solvent of these two solvents is added. Step (c): τ of the crystalline tenofovirodisoxyl free base by recrystallization = for crystallization of the result of steps (a)-(b). Violent stirring and optionally seeding as necessary yields a crystalline tenofovirisotropoxyl freebase.

It is advantageous to stir vigorously the tenofovirisopropyl radical and the solvent, the stirring time being 1-8 hours, more specifically 1-4 hours.

Upon stirring, seeding of the tenofovirosophyxyl crystalline free base according to the present invention may be performed seeding to speed up the precipitation time of the crystal. Seeding is not essential but is effective when no crystalline solid is precipitated after several hours of stirring. According to another aspect of the present invention, the present invention provides a pharmaceutical composition for anti-viral comprising the above-mentioned crystalline tenofovirodisofoxyl free base of the present invention.

According to another embodiment of the invention, the virus is human immunodeficiency virus (HIV) -1 or hepatitis B virus (HBV).

Therefore, the composition of the present invention is a drug useful for the treatment of HIV-1 infection and chronic hepatitis B. When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in the preparation, lactose, dextrose, sucrose, sorbbi, manny, starch, acacia rubber, calcium phosphate, alginate, Gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil Including, but not limited to. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives ^, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered to various mammals such as oral or parenteral to mammals such as rats, mice, livestock, humans, and in the case of parenteral administration, intravenous injection, subcutaneous injection, intramuscular injection, abdominal cavity Administration by infusion, transdermal administration, or the like.

Suitable dosages of the pharmaceutical compositions of the present invention will vary depending on factors such as the formulation method, mode of administration, age, weight, sex, pathological condition, food, time of administration, route of administration, rate of excretion and reaction in response to the patient. It may be prescribed. The daily dose of the pharmaceutical composition of the present invention is, for example, 0.01-100 mg / kg. The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be used in any form suitable for pharmaceutical preparations, including powders, granules, tablets, capsules, suspensions, oral formulations such as emulsions, syrups, aerosols, external preparation suppositories such as ointments and creams, and sterile injectable solutions. It may further comprise a dispersant or stabilizer. 【Effects of the Invention】

The features and advantages of the present invention are summarized as follows: (a) The tenofovirosophyxyl free base according to the present invention can minimize the generation of a flexible material with the change over time, compared to tenofovirisoofoxyl fumarate, lowering the amount of impurities generated during the storage of the product, the stability of the formulation Can increase.

(b) In addition, the tenofovirosophyloxy free base according to the present invention can be produced with much better Sejo purity (pur i ty) compared to the known tenofovirisophyxyl free base.

(c) In addition, the tenofovirodisoxyl free base according to the present invention has excellent physicochemical properties such as hygroscopicity, solubility, and adhesiveness, so that it can be used as a useful active ingredient of a pharmaceutical composition.

(d) Tenofovirosophyxyl free base according to the present invention is industrially useful because it is suitable for high yield and high purity, the specification of the flexible material and residual solvent as a pharmaceutical raw material. [Brief Description of Drawings]

1 is a view showing an X-ray diffraction pattern of the tenofovirosophyxyl free base crystal form according to the present invention.

FIG. 2 is a diagram showing a DSC (Di f ferent i Al scanni ng cal or imet ry) record of the tenofovirisoproxyl free base crystal form according to the present invention.

Fig. 3 shows the X-ray diffraction pattern (top) of the tenofovirosophyxyl free base crystalline form of the present invention and the X-ray diffraction pattern (bottom) of the glass base prepared by the method described in Korean Laid-Open Patent Publication No. 10-2014-0028790. Are diagrams comparing each.

FIG. 4 is a diagram showing the DSC data of tenofovirosophyloxy free base crystalline form (green graph) and tenofovirosophyloxy free base (blue graph) of JP 10-2014-0028790 of the present invention. .

5 is a view showing a particle diagram of the tenofovirosophyxyl free base of the present invention.

6 is a view showing a particle diagram of a free base described in Patent Publication No. 10—2014-0028790. Figure 7 is a view showing a comparison of the powder picture of the free base (left) and the tenofovirsoxotoxyl free base of the present invention (right) according to the invention described in Korea Patent Publication No. 10-2014-0028790. While the glass base according to Korean Patent Laid-Open Publication No. 10-2014- 0028790 is strong and agglomerated, the tenofoviroxoxyl free base according to the present invention shows the form of a homogeneous powder.

[Specific contents to carry out invention]

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. . In the example, the 1H- 蘭 R analytical instrument used Agilent 400 MHz, HPLC used UltiMate-3000 from Dionex, melting point was DSC 823 from Met tier Toledo and B-545 from Buchi, Reagents and solvents were purchased from Aldrich unless otherwise noted. Example

Example 1 Tenofovirosophyroxyl Free Base Novel Crystalline Production Phase 200 g of tenofovirisofluorofumalate (China Asta chemical) was dissolved in 3.6 L of methylene chloride and 360 ml of methanol. An aqueous 10% sodium carbonate solution was added thereto, stirred for 10 minutes, the pH was adjusted to about 9-10, and the organic layer was separated. Magnesium sulfate was used to remove water from the organic worms, and the filtrate was concentrated under reduced pressure at 45 ^° C. Thereafter, 600 ml of isopropyl alcohol was added thereto, followed by stirring at 24 ° C. for 2 hours, followed by filtration (washing with isopropyl alcohol 200 ml) to obtain 140 g of tenofovirodisopropyl base crystalline form (melting point (nip): 106 ^° C.). Purity: 99.90%).

DMS0-d6 8.133 (1H, s), 8.026 (1H, s), 7.192 (2H, s), 5.577-5.506 (4H, m), 4.840-4.803 (2H, m), 4.20 (1H, d, J = 2 Hz), 4.194 (1H, d, J = 6 Hz), 3.999-3.953 (3H, m), 1.246 (12H, d, J = 6H), 1.070 (3H, d J = 6Hz). Example 2 Tenofovirosophyxyl free base In a novel crystalline form, 200 g of tenofovirisophthoxyfumalate was dissolved in 3.6 L of methylene chloride and 360 ml of methanol. Thereafter, 10% aqueous sodium carbonate solution was added thereto, stirred for 10 minutes, the pH was adjusted to about 9-10, and the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, filtered and concentrated under reduced pressure at 45 ^° C. Thereafter, 300 ml of isopropyl alcohol and 300 ml of ethanol were added thereto, followed by stirring at 24 ^° C. for 2 hours, followed by filtration (washing with 200 ml of isopropyl alcohol) to obtain 135 g of a new crystalline form of tenofovirodisopropyl base (melting point (mp): 106 ^° C, Purity: 99.90%).

DMS0-d6 8.133 (1H, s), 8.026 (1H, s), 7.192 (2H, s), 5.577-5.506 (4H, m), 4.840-4.803 (2H, m), 4.20 (1H, d, J = 2 Hz), 4.194 (1H, d, J = 6 Hz), 3.999-3.953 (3H, m), 1.246 (12H, d, J = 6H), 1.070 (3H, d, J = 6 Hz). Example 3: Manufacture of Tenofovirosophyxyl Free Base Novel Crystalline At room temperature, 200 g of tenofovirisoxofulmulfate was dissolved in 3.6 L of methylene chloride and 360 ml of methanol. Thereafter, an aqueous 10% sodium carbonate solution was added thereto, stirred for 10 minutes, the pH was adjusted to about 9-10, and the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, filtered and concentrated under reduced pressure at 45 ^° C. Thereafter, 600 ml of ethane was added thereto, followed by stirring at 24 ^° C. for 2 hours, followed by filtration (washing with 200 ml of isopropyl alcohol) to obtain 135 g of a new crystalline form of tenofovirisoxopropyl base (melting point (mp): 106 ^° C ᅳ). Purity: 99.88%).

DMS0-d6 8.133 (1H, s), 8.026 (1H, s), 7.192 (2H, s), 5.577-5.506 (4H, m), 4.840-4.803 (2H, m), 4.20 (1H, d, J = 2 Hz), 4.194 (1H, d, J = 6 Hz), 3.999-3.953 (3H, ni), 1.246 (12H, d ᅳ J = 6H), 1.070 (3H, d, J = 6 Hz). Example 4 Preparation of New Crystalline Tenofovirosophyxyl Free Base

In the phase, 100 g of tenofovirisonafadasilate was dissolved in 1.8 L of methylene chloride and 180 ml of methanol. Thereafter, an aqueous 10% sodium carbonate solution was added thereto, stirred for 10 minutes, the pH was adjusted to about 9-10, and the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, filtered, and concentrated under reduced pressure at 45 degrees. Thereafter, 300 ml of ethanol was added, followed by stirring at 24 ° C. for 2 hours, followed by filtration (washing with 200 ml of isopropyl alcohol) to give 56.5 g of a new crystalline form of tenofoviridisopropyl base (melting point (mp): 106 ° C, purity ( Purity): 99.80%).

DMS0-d6 8.133 (1H, s), 8.026 (1H, s), 7.192 (2H, s), 5.577-5.506

(4H, m), 4.840-4.803 (2H, πι), 4.20 (1H, d, J = 2Hz), 4.194 (1H, d, J = 6Hz), 3.999-3.953 (3H, m), 1.246 (12H, d, J = 6H), 1.070 (3H, d, J-6 Hz). Example 5 Tenofovirosophyloxy Free Base New Crystalline Preparation

400 ml of isopropyl alcohol was added to 100 g of liquid tenofovirisopropyl-based free base prepared by the method of U.S. Patent No. 5935946, followed by vigorous stirring at room temperature for 4 hours, and then the resulting solid was filtered (isopropyl alcohol). Washed with 200 ml) 92.5 g of a new crystalline form of tenofovirisophthoxyl free base was obtained (melting point (mp): 106 ° C, Purity: 99.89%).

DMS0-d6 8.133 (IH, s), 8.026 (1H, s), 7.192 (2H, s), 5.577-5.506 (4H, m), 4.840- .803 (2H, m), 4.20 (1H, d, J = 2 Hz), 4.194 (1H, d, J = 6 Hz), 3.999-3.953 (3H, m), 1.246 (12H, d, J = 6H), 1.070 (3H, d, J = 6 Hz). Example 6 Manufacture of Tenofovirisophthoxyl Free Base New Crystalline Form

In phase silver, 100 g of tenofovirdisoficylpydolate was dissolved by adding 1.8 L of methylene chloride and 180 ml of methanol. Thereafter, the prepared 10% aqueous sodium carbonate solution was added thereto, stirred for 10 minutes, the pH was adjusted to about 9-10, and the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, filtered and concentrated under reduced pressure at 45 ° C. After that 300 ml of ethane was added, followed by stirring at 24 ^° C. for 2 hours, followed by filtration (washing with 200 ml of isopropyl alcohol) to give 61 g of tenofovirodisoftosyl base novel crystalline form (melting point (nip): 105.5 ^° C.). Purity: 99.90%).

DMS0-d6 8.133 (1H, s), 8.026 (1H, s), 7.192 (2H, s), 5.577-5.506 (4H, in), 4.840-4.803 (2H, m), 4.20 (1H, d, J = 2 Hz), 4.194 (1H, d, J = 6 Hz), 3.999-3.953 (3H, m), 1.246 (12H, d, J = 6H), 1.070 (3H, d, J = 6 Hz). Example 7: Preparation of a new crystalline form of tenofovirisoxosiloxane free base 100 g of the free base obtained by the method known in Korean Patent Publication No. 10-2014-0028790 was added 300 ml of isopropyl alcohol and 300 ml of ethanol, and then at 24 ^° C. Stir vigorously for 4 hours, and then filter (wash 200 ml of isopropyl alcohol) to give 95 g of tenofovirodisoxyl base novel crystalline form (melting point (mp): 106 ^° C., Purity: 99.93%).

DMS0-d6 8.133 (1H, s), 8.026 (1H, s), 7.192 (2H, s), 5.577-5.506

(4H, m), 4.840-4.803 (2H, m), 4.20 (1H, d, J = 2 Hz), 4.194 (1H, d, J = 6 Hz), 3.999-3.953 (3H, m), 1.246 (12H, d, J = 6H), 1.070 (3H, d, J-6 Hz). Comparative Example 1: Preparation of the free base of the method known in Korea Patent Publication No. 10-2014-0028790

200 g of tenofovirodisoxylfumalate was obtained using a method known from Korean Patent Application Publication No. 10-2014-0028790 to obtain 125 g of free base (melting point (mp): 96), purity: 98.37% ).

Experimental Example 1 Experimental Example 1 Polymorphic Formation (XRD) of Novel Tenofovirosophyxyl Free Base

X-ray powder diffraction pattern is widely used to differentiate crystalline forms and hydrates of a unique characteristic of the ^drug. Therefore, the X-ray powder pattern of the tenofovirisophyxyl free base of this invention was measured. Powder X-ray diffraction (PXRD) analysis of a new crystalline form of tenofobicisoproxyl free base showed that the 2Θ diffraction angle was 7.4 ± 0.2 ^° , 8.1 ± 0.2 ^° , 10.8 ± 0.2 ^° , 13 ^° 0.2 ^° , 14.4 ± 0.2 °, 14.8 ± 0.2 ^° , 16.2 ± 0.2 ^° , 16.7 ± 0.2 °, 17.6 ± 0.2 °, 18.04 ± 0.2 °, 19.19 ± 0.2 °, 19.4 ± 0.2 °, 20.9 ± 0.2 °, 22.9 ± 0.2 °, 24.4 ± 0.2 ^° , Specific peaks were obtained at 26.7 土 0.2 ^° and 28.6 ± 0.2 ^° , respectively.

Scanning range: 5 to 40 °, 2Θ

Scan Speed: 6 deg / min

The value of the powder X-ray diffraction (PXRD) angle of the novel tenofovirisoxoyl free base crystalline form shows a diffraction angle different from that of the tenofovirosomal free base of US Patent Publication No. 2011/0112292. Bar, it was confirmed that it is a new crystalline form.

In addition, a pattern of diffraction angles different from that of the free base prepared in Korean Patent Laid-Open Publication No. 10-2014-0028790 was shown. The difference is shown in FIG. 3 as comparative experimental data.

As shown in Fig. 3, the upper graph is the tenofovirisoproxyl free base of the present invention, and the lower graph is the data of the free base described in Korean Laid-Open Patent Publication No. 10-2014-0028790. . Experimental Example 2 Analysis of Differential Scanning Calorimetry (DSC) of a Novel Tenofobidisoproxyl Free Base

Temperature Differential Scanning Calorimetry of the New Tenofovirisoopoxyl Free Base Novel Crystalline Form Presented in the Present Invention for Comparison with Novel Tenofovirosophyloxy Free Bases Having Melting Points of 61 ^° C to 66 ^° C as disclosed in US2011 / 0112292 (DSC) analysis showed that the endothermic start temperature 103 ± 3 ^° C, endothermic silver

107 土; c shows an endothermic peak.

Temperature range: 20-150 ^° C

Temperature rising rate: 10 ^° C / min The new crystalline form of tenofovirisoxoyl free base presented in the present invention is

Thermodynamically ^' stable because it has a melting point above 100 ^° C, It is also good to use as an active ingredient of medicines, there is also an advantage in the preparation of acid addition salts, such as tenofovirisoproxyl.

On the other hand, DSC measurement results of the salt prepared in Korean Patent Application Laid-Open No. 10-2014-0028790 were 95 degrees and the endothermic peak was different from the crystalline form of the present invention and was not a thermodynamically stable crystalline form.

The DSC data of the novel crystalline form of the tenofovirosophyloxy free base of the present invention (green graph) and the tenofovirosophyloxy free base (blue graph) of JP-A 10-2014—0028790 is shown in FIG. 4. Experimental Example 3 ： Stability Comparison Evaluation (Acceleration / Severity)

The stability test of a drug is to set the storage period and the period of use of the drug, and to set the expiration date by determining the significant change based on the specified test method after setting the appropriate standard. Is one of the most important factors in the commercialization of drugs.

Therefore, in order to confirm the feasibility of commercialization of tenofovirisoxopropyl free base having a novel crystalline form of the present invention, ICH guide using crystalline tenofovirisoxofucil fumaric acid (Ast a chem i ca ls, ch i na) as a control Acceleration and harsh stability were performed along the line and analyzed using Liquid Chromatography (HPLC) analysis described in the USP. The results are shown in Tables 1 and 2.

Table 11

Adenine 0.16 ― ― _ ― _ 0.003 0.006 0.006 tenofovir 0.24 0.217 0.317 0.356 0.256 0.776 0.978 disoproxyl

Monoester

Tenofovir 0.80 0.052 0.082 0.231 0.005 0.00S 0.010 0.025 0.036 0.046 disoproxyl

Ethyl ester

Tenofovir 0.82 0.103 0.011 0.015 0.078 disoproxyl

Isoester

Tenofovir 1.40 0.026 0.035 0.037 0.022 0.023 0.029 0.010 0.025 0.036 disoproxyl

Carbamate

Tenofovir 1.76 0.26 0.28 Disoftoxyl

Dimer

Table 2

Severe (60。 C ± 2 ^° C) condition

Ethyl ester

Tenofovir 0.82 0 .125 0.001 0.001 0.003 0.011 0.019 0.030 disoproxyl

Isoester

Tenofovir 1 .40 0.026 0.050 0.053 0.022 0.024 0.024 0.010 0.035 0.356 Disoftoxyl

Carbamate

Tenofovir 1.76 0.037 0.056 disoproxyl

Dimer As shown in Table 1 and Table 2, the tenofovirisotropylic free base having the novel crystalline form represented by the present invention is a tenophoreodisopropyl fumaric acid or a control material under accelerated and harsh conditions. It can be seen that the material is very stable compared to the free base specified in 10-2014—0028790. In particular, the number of flexible materials and the increasing width is small, there is no problem to be affected by decomposition products that may occur during storage. In particular, Tenofovirisopromonol ester (Tenofov irl sopropox il monoester), which is the main product of tenofovirisofluorofumarate, was 1.246% in case of tenofovirisofluorofumaric acid under severe conditions. In the case of the free base specified in Patent Publication No. 10-2014-0028790, the increase in the amount of the decomposition product is 0.1% or less. Significantly improved.

Therefore, it can be seen that tenofovirosophyloxy free base according to the present invention can maintain a high purity for a long time by minimizing the generation of the flexible material, it is excellent storage stability material. Experimental Example 4: Evaluation of Solubility Comparison According to pH A complete solution was prepared for each pH as shown in Table 3, and each sample was filtered at 100 ° C. at a stirring speed of 25 ° 150 rpm at each pH, and then filtered for each sample to remove undissolved substances. Quantification using a chromatography (HPLC) assay.

Solubility for each pH was determined based on the solubility obtained under the above conditions.

[Table 3]

As shown in Table 3, the novel crystalline tenofovirisoproxyl free base made in the present invention has an equivalent degree of solubility in buffer solution for each pH compared to tenofovirodisoxylfumaric acid. Therefore, it can be seen that there is no difference in solubility according to the silvering effect between tenofovirisofumaric acid and tenofovirisofluorosilic acid base of the present invention. Experimental Example 5 Comparative Evaluation of Hygroscopicity and Particle Size

Since hygroscopicity is one of the important factors for processing and storage of the physical properties required for use as a drug substance, the degree of hygroscopicity was compared with respect to the novel crystalline tenofovirisoxosiloxane base. Experimental method was used as a control group of tenofovirdifuxyl fumarate 500 mg and 500 mg of free base of Patent Publication No. 10-2014-0028790, and the new crystal tenofovirdiisopropyl glass base 500 mg of glass tube for measuring hygroscopicity After drying in nitrogen at 25 ° C for 12 hours, the rate of mass change of the sample was automatically measured at a relative humidity of 15, 35, 55, 75, and 95% using a hygroscopic measuring instrument (Quant achrome). Measured as shown in Table 4. According to the Korea Food and Drug Administration standard, tenofovirisophthoxyfumarate is controlled to less than 1.0% moisture. As shown in Table 4, tenofovirdiphyxyl fumarate and the free base of Korean Patent Application Laid-open No. 10-2014-0028790 increase the moisture content rapidly as the relative humidity increases, but the novel crystalline tenofobibi according to the present invention. Whereofoxyl free base has a relatively low hygroscopicity can be seen in the form of pharmaceutically very stable crystals.

Table 4

In addition, in the case of a solid powder, compatibility with an excipient during tableting, a flow chart of particles, and uniformity of particles are very important. This is how homogeneous the particles are distributed on the particle distribution map, and in general, the ideally shaped powder having a very uniform and symmetrical particle distribution on the PSD is known to be easy to tablet. The particle size of the free base of Unexamined-Japanese-Patent No. 10-2014-0028790 was measured and compared. Particle size was measured using a Mast sizer 2000 (niasters i zer 2000) equipment using a general solid dispersion method. 5 and 6 show particle sizes of the novel crystalline tenofovirosophyxyl free base and Patent Publication No. 10-2014—0028790, respectively. As shown in FIGS. 5 and 6, the novel crystalline tenofovirodisoxyl glass base of the present invention shows a symmetrical particle distribution form with a narrow and narrow distribution, but the glass of Korean Patent Application Laid-Open No. 10-2014-0028790 In the case of a base, it has a particle distribution in the form of a pharmaceutical formulation with a wide variety of peaks.

In addition, as shown in Figure 7, the glass base (left) according to Korean Patent Laid-Open No. 10-2014-0028790 has a strong adhesiveness and agglomeration, while the tenofovirsotoxyl glass base according to the present invention (right) ) Shows the form of a homogeneous powder. Having described the specific part of the present invention in detail, it will be apparent to those skilled in the art that the specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the invention will be defined by the appended claims and equivalents thereof.

Claims

[Patent Claims]

[Claim 1]

Specific peak diffraction angle (2Θ ± 0.1 ^° ) in X-ray diffraction (PXRD) '7.4 ± 0.2 ^° , 8.1 ± 0.2 ^° , 10.8 ± 0.2 ^° 13 ± 0.2 ^° 14.4 土 0.2 ° 14.8 ± 0.2 ^° , 16.2 ± 0.2 ^° 16.7 ± 0.2 ° 17.6 ± 0.2 ^° , 18.04 ± 0.2 ° 19.19 ± 0.2 ^° 19.4 士 0.2 °, 20.9 ± 0.2 ^° 22.9 ± 0.2 ^° , 24.4 ± 0.2 ^°

A crystalline tenofovirisopropyl radical represented by the following Chemical Formula 2, characterized by having a melting point in the range of 26.7 ^{° C} 0.2 ^° and 28.6 ± 0.2 ^° and having a differential scanning calorimetry (DSC) characteristic of 107 ± 3 t:

Formula 2

[Claim 2]

The crystalline tenofovirisophthoxyl free base according to claim 1, wherein the crystalline tenofovirisophthosil free base has the X-ray diffraction pattern shown in FIG.

[Claim 3]

Process for the preparation of crystalline tenofovirisotropylic free base comprising the following steps:

(b) adding magnesium sulfate (MgSO ₄ ) or sodium sulfate (Na ₂ SO ₄ ) as an absorbent and stirring the resultant of step (a); (c) filtering the resultant of step (b) and concentrating and drying under reduced pressure; And

(d) ethanol and isopropane in the resultant of step (c); Or adding a co-solvent of ethanol and isopropanol, and obtaining crystalline tenofovirisoxosiloxane free base through vigorous stirring and selective seeding.

[Claim 4]

(a) ethane and isopropane in the slime free, amorphous free or crystalline free base of tenofovirisoxo; Or adding a co-solvent of ethanol and isopropanol;

(b) filtering the product of step (a);

(c) obtaining the crystalline tenofovirosophyxyl free base through selective seeding of the resultant of step (b).

[Claim 5]

The method of claim 3, wherein the adjustment of the pH of step (a) is characterized in that it is adjusted to a pH range of pH 9 to 10.

[Claim 6]

According to claim 3, Ethanol, isopropanol of step (d); Or ethane and a co-solvent of isopropanol in the range of 10-50 ml for tenofoviroxy free base lg.

[Claim 7]

The method of claim 3, wherein the stirring time for crystallization of step (d) is 1 to 8 hours. [Claim 8]

A pharmaceutical composition for an antiviral comprising the crystalline tenofovirodisoxyl free base of claim 1.