KR20070111099A - Novel crystalline form of sitagliptin hydrochloride - Google Patents

Abstract

A novel crystalline form of sitagliptin hydrochloride having inhibitory effects on DP-IV(dipeptidyl peptidase-IV) is provided to improve water stability by reducing water-absorption over the time, thereby inhibiting decomposition and purity lowering of the compound and storing it for a long period. A novel crystalline form of sitagliptin hydrochloride having a powder X-ray diffraction pattern showing peaks at 2-theta of 13.7, 18.0, 22.6, 25.7 and 27.0±0.2 degrees is prepared by reacting (a) amine hydrochloride or a mixture of amine compounds and hydrochloric acid having smaller pKa than maximum pKa of sitagliptin with (b) sitagliptin in at least one organic solvent selected from hydroxy, nitroalkane, acetonitrile, acetate, ether, alkane chloride and ketone, and optionally stirring the reaction product to promote crystallization of sitagliptin.

Description

Novel crystalline form of cytagliptin hydrochloride, preparation method thereof and pharmaceutical composition comprising the same {NOVEL CRYSTALLINE FORM OF SITAGLIPTIN HYDROCHLORIDE}

1 is a view showing a powder X-ray diffraction pattern for the crystalline form of cytagliptin hydrochloride according to the present invention,

Figure 2 is a characteristic DSC thermal analysis for the crystal form of cytagliptin hydrochloride according to the present invention,

3 is a diagram showing an IR spectrum of the crystalline form of cytagliptin hydrochloride according to the present invention.

4 is a diagram showing a powder X-ray diffraction pattern for the comparative cytagliptin hydrochloride (as disclosed in WO 03/004498) prepared according to the prior art,

FIG. 5 is a characteristic DSC thermal analysis of the comparative cytagliptin hydrochloride (as disclosed in WO 03/004498) prepared according to the prior art,

FIG. 6 is a diagram showing an IR spectrum of a comparative cytagliptin hydrochloride (as disclosed in WO 03/004498) prepared according to the prior art.

The present invention relates to a novel crystalline form of cytagliptin hydrochloride with improved water stability, a preparation method thereof, and a pharmaceutical composition comprising the same.

Diabetes is generally known in two forms. Type 1 diabetes, namely insulin dependent diabetes mellitus, develops by producing little or no insulin, a hormone that regulates sugar utilization in the body. In contrast, type 2 diabetes, namely insulin independent diabetes mellitus, has sufficient insulin in the body, but has been shown to have an effect on the insulin-promoting effect on sugar and lipid metabolism in major insulin-sensitive tissues such as muscle, liver and fat cells. With greater tolerance, plasma insulin levels develop by failing to overcome significant resistance.

Among the two types of diabetes, various treatment methods for type 1 diabetes have been researched and developed in various ways, but the treatment for type 2 diabetes has been limited for a long time.

For this reason, various drugs are being studied to help treat or prevent type 2 diabetes, among which dipeptidyl peptidase-VI (“DP-VI” or “DPP-VI”; hereinafter “DP-VI”). Compounds that are inhibitors of enzymes continue to be studied as drugs useful in the treatment of type 2 diabetes. The usefulness of such DP-VI inhibitors for the treatment of type 2 diabetes is that glucagon-like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP), which are a type of incretin produced when food is consumed in the body, may be The VI inhibitor is inactivated. That is, the incretin promotes insulin production, whereby DP-VI inhibitors induce reduced inactivation of this incretin, whereby the incretin is produced by the body only when food is consumed. Because of this, by using the DP-VI inhibitor, an increase in the level of insulin at an inappropriate time, such as between meals, can be suppressed, thereby increasing the risk of hypoglycemia, a dangerous side effect associated with the use of insulin secretagogues. You can expect to increase insulin without.

On the other hand, WO 03/004498 (published Jan. 16, 2003) of Merck & Co., Ltd. has a strong DP-VI inhibitory activity, and is a compound useful for the treatment of type 2 diabetes. 7-[(3R) -3-amino-4- (2,4,5-trifluorophenyl) butanoyl] -3- (trifluoromethyl) -5,6,7,8-tetrahydro-1 , 2,4-triazole- [4,3-a] pyrazine (also referred to as "SITAGLIPTIN"; hereafter abbreviated as "cytagliptin") has been disclosed.

[Formula 1]

In addition, WO 03/004498 discloses a method for preparing hydrochloride salt of cytagliptin of Chemical Formula 1 (Example 7), and also treats type 2 diabetes comprising such cytagliptin hydrochloride as an active ingredient. For pharmaceutical compositions are disclosed.

However, the cytagliptin hydrochloride disclosed in WO 03/004498 has very high hygroscopicity in air. For this reason, when the pharmaceutical composition and the medicament for treating type 2 diabetes are prepared using the same, decomposition of the active ingredient occurs due to the water absorption of the active ingredient, ie, cytagliptin hydrochloride, during storage of such a drug. For this reason, the drug containing the sitagliptin hydrochloride disclosed in the prior art as an active ingredient has a problem that the purity of the active ingredient rapidly decreases with time and can not be stored while maintaining its efficacy for a long time. Furthermore, the citagliptin hydrochloride disclosed in the prior art has been found to be unsuitable for use as an active ingredient in pharmaceutical compositions and pharmaceuticals for treating type 2 diabetes due to the large hygroscopicity described above.

WO 05/030127 (published Apr. 7, 2005) of Merck & Company, Inc., solves the problem of the sitagliptin hydrochloride disclosed in WO 03/004498 to some extent, and is a cytagliptin phosphate anhydride. Form IV of was disclosed. However, the crystal form IV of the cytagliptin phosphate anhydride also has a relatively high hygroscopicity and it is described in the examples of WO 05/030127 above that it absorbs moisture in relatively high humidity air and converts it to monohydrate. Therefore, Form IV of the cytagliptin phosphate anhydride also cannot completely solve the problem due to the large hygroscopicity exhibited by the cytagliptin hydrochloride.

Furthermore, since Form IV of cytagliptin phosphate anhydride should be prepared once from cytagliptin phosphate monohydrate and then subjected to a separate process therefor, the overall manufacturing process is complicated and the yield accordingly There was also a problem of poor performance.

Accordingly, the present invention is to provide a novel crystalline form of cytagliptin hydrochloride with improved water stability by solving the above problems of the prior art.

Another object of the present invention is to provide a simple method for preparing a novel crystalline form of the cytagliptin hydrochloride.

Another object of the present invention is to provide a pharmaceutical composition comprising the novel crystalline form of the cytagliptin hydrochloride.

In order to achieve the above object, the present invention provides a crystal form of cytagliptin hydrochloride having a powder X-ray diffraction pattern exhibiting peaks at 2θ of 13.7, 18.0, 22.6, 25.7 and 27.0 ° ± 0.2 °. Preferably, the crystal form of the stagliptin hydrochloride according to the present invention is 6.5, 8.0, 13.7, 16.0, 18.0, 18.7, 19.6, 19.8, 20.3, 22.6, 23.4, 24.8, 25.4, 25.7, 27.0, 29.4, 32.3 ° ± It may have a powder X-ray diffraction pattern showing a peak at 2θ of 0.2 °, more preferably, 6.3, 6.5, 7.6, 8.0, 13.2, 13.7, 16.0, 18.0, 18.7, 19.6, 19.8, 20.3, 21.7, 22.6, 23.4, 23.8, 24.8, 25.4, 25.7, 26.5, 27.0, 27.6, 28.8, 29.4, 29.8, 31.0, 31.6, 32.0, 32.3, 32.9, 33.3, 34.2, 34.8, 35.6, 36.0, 36.5, 37.5, 38.3, It may have a powder X-ray diffraction pattern showing a peak at 2θ of 38.7, 43.3 ° ± 0.2 °. Also, most preferably, the crystalline form of the stagliptin hydrochloride according to the present invention may have a powder X-ray diffraction pattern shown in FIG. 1.

As will be described in more detail in Test Example 1 to be described later, the present inventors through the powder X-ray diffraction pattern analysis, the novel crystalline form of the cytagliptin hydrochloride according to the present invention (see powder X-ray diffraction pattern of Figure 1) WO 03 It was confirmed that it had a specific crystal form that was completely different from the cytagliptin hydrochloride disclosed in US00 / 004498 (cytagliptin hydrochloride disclosed in Example 7 of WO 03/004498, etc .; see the powder X-ray diffraction pattern of FIG. 4).

In addition, the novel crystalline form of the cytagliptin hydrochloride according to the present invention can also be confirmed by thermal properties. Referring to the DSC thermal analysis of FIG. 2, the novel crystalline form of cytagliptin hydrochloride according to the invention at 10 ° C./min shows a clear endothermic peak at 135.08 ° C. FIG. In comparison, the cytagliptin hydrochloride disclosed in WO 03/004498 only exhibits an exothermic peak at 123.33 ° C. This means that while the cytagliptin hydrochloride disclosed in WO 03/004498 does not exhibit a specific crystalline form and is almost amorphous, the novel crystalline form of cytagliptin hydrochloride according to the present invention has a specific single crystalline form.

In addition, the novel crystalline form of the cytagliptin hydrochloride according to the present invention can also be identified through the IR spectrum. Referring to the IR spectrum of FIG. 3, the novel crystalline form of cytagliptin hydrochloride according to the invention exhibits an IR spectrum which is distinct from the cytagliptin hydrochloride (see IR spectrum of FIG. 6) disclosed in WO 03/004498. It is confirmed that it possesses a specific crystalline form that distinguishes it from the cytagliptin hydrochloride disclosed in WO 03/004498.

On the other hand, in general, it is well known to those skilled in the art that the biological or pharmaceutical activity of compounds having a polymorphic or amorphous form is the same or equivalent to the same compounds having a particular crystalline form. However, in WO 03/004498, it is apparent to those skilled in the art that cytagliptin hydrochloride exhibits the activity as a DP-IV inhibitor and thus shows a pharmacological activity useful for the treatment or prevention of type 2 diabetes. Thus, the specific crystalline form of the sitagliptin hydrochloride according to the present invention, which differs only in the crystal form from the sitagliptin hydrochloride disclosed in WO 03/004498, is also the same or equivalent DP to the sitagliptin hydrochloride disclosed in WO 03/004498. It can be obvious to those skilled in the art that it shows activity as a -IV inhibitor and thus shows a pharmaceutical activity useful for the treatment or prevention of type 2 diabetes.

In addition, according to the results of the inventors' experiments, the specific crystalline form of the cytagliptin hydrochloride according to the present invention is the cytagliptin hydrochloride disclosed in WO 03/004498 or cytagliptin phosphate anhydride disclosed in WO 05/030127. It was confirmed that it exhibits the activity as a DPP-IV inhibitor equivalent to Form IV of (see Test Example 2).

In addition, as will be described in more detail through Test Example 3 to be described later, the results of the inventors of the present inventors, surprisingly, the specific crystalline form of the cytagliptin hydrochloride according to the present invention as described above does not substantially exhibit hygroscopicity in the air time It has been confirmed that high stability over time, accordingly, can maintain high purity and efficacy for a long time. More specifically, the specific crystalline form of the cytagliptin hydrochloride according to the present invention exhibits hygroscopicity of only about 1/20 of the cytagliptin hydrochloride disclosed in WO 03/004498, and the cytagliptin disclosed in WO 05/030127. It was confirmed that the hygroscopicity was only about 1 / 7.5 compared to Form IV of phosphate anhydride.

Therefore, by using the specific crystalline form of the cytagliptin hydrochloride according to the present invention as the active ingredient, it is possible to maintain the high purity of the active ingredient for a long time as the decomposition of the active ingredient by the absorption of moisture in the air is minimized, thereby It is evident that it is possible to provide a medicament for the treatment or prophylaxis of type 2 diabetes which can maintain a high medicament for a long time.

On the other hand, the present invention also under any one or two or more organic solvents selected from the group consisting of hydroxy solvent, nitroalkane solvent, acetonitrile solvent, acetate solvent, ether solvent, methane chloride solvent and ketone solvent, a) a hydrochloride or mixture of an amine compound and hydrochloric acid of an amine compound having a pKa less than the maximum pKa of cytagliptin, and b) reacting cytagliptin according to the present invention. Provided are methods for preparing the crystalline form.

Regarding this preparation method, WO 97/07112 (published Feb. 27, 1997) discloses the reaction of free amine compounds with donor compounds having amine salt moieties having a pKa less than the maximum pKa of such free amine compounds. , A method for producing an acid addition salt of the free amine compound is disclosed. Referring to WO 97/07112 above, as a result of this reaction, the acid moiety of the donor compound is converted into the free amine compound to prepare an acid addition salt of the free compound, and in the course of this reaction, the polarity of the organic solvent is reduced. By adjusting, acid addition salts of the free compounds can be crystallized into specific crystalline forms.

However, WO 97/07112 does not disclose or anticipate the fact that such preparation methods can be used to prepare cytagliptin hydrochloride or its specific crystalline form.

In the present invention, the application of the acid-moiety conversion reaction disclosed in WO 97/07112 is prepared to produce cytagliptin hydrochloride, and in this preparation process, an organic solvent having an appropriate polarity, that is, a hydroxy solvent, a nitroalkane solvent, By using any one or two or more organic solvents selected from the group consisting of acetonitrile solvents, acetate solvents, ether solvents, alkane chloride solvents and ketone solvents, a specific crystalline form of cytagliptin hydrochloride according to the present invention is prepared. do.

At this time, in the method for producing a crystal form of cytagliptin hydrochloride according to the present invention, b) cytagliptin is a) hydrochloride or amine compound of the amine compound having a pKa less than the maximum pKa of the cytagliptin, and the hydrochloric acid And reacted with the mixture, and as a result of the measurement of the inventors, the maximum pKa of b) cytagliptin was found to be about 8.5-9.

Accordingly, a) a hydrochloride or amine-based compound of an amine-based compound having a pKa less than the maximum pKa of cytagliptin, and a mixture of hydrochloric acid, a hydrochloride of an amine-based compound having a pKa of less than about 8.5-9 or such an amine-based compound. Mixtures of compounds and hydrochloric acid (or hydrogen chloride) can be used. For example, hydrochlorides of unrestricted amine compounds such as pyridine compounds, quinoline compounds and aniline compounds having a pKa of less than about 8.5-9 or mixtures of these amine compounds with hydrochloric acid (or hydrogen chloride) can be used. have. Preferably, a) a hydrochloride or mixture of an amine compound and hydrochloric acid of an amine compound having a pKa less than the maximum pKa of cytagliptin, a mixture of hydrochloride salt of triallineamine, hydrochloride salt of pyridine, triallinamine and hydrochloric acid. Or mixtures of pyridine and hydrochloric acid can be used.

On the other hand, the method for producing a crystal form of cytagliptin hydrochloride according to the present invention, after the reaction step of a) a mixture of the hydrochloride or amine compound and hydrochloric acid of the amine compound and b) cytagliptin, stirring the reaction product It is preferable to further include the step of promoting the crystallization of the hydrochloride salt of cytagliptin.

Since the specific crystalline form of cytagliptin hydrochloride according to the present invention has great stability, simply proceeding with the reaction step of a) a hydrochloride of an amine compound or a mixture of an amine compound and hydrochloric acid and b) cytagliptin Although it can be prepared, by further stirring the reaction product, the crystallization of the cytagliptin hydrochloride is further promoted to prepare a specific crystalline form of the cytagliptin hydrochloride according to the present invention in high yield and high purity within a short time. can do.

In the method for producing a crystal form of cytagliptin hydrochloride according to the present invention, polarity is appropriately adjusted in the course of the reaction of a) the hydrochloride or the amine compound and the hydrochloric acid of the amine compound and b) cytagliptin. By causing the crystallization of cytagliptin hydrochloride, certain crystalline forms of cytagliptin hydrochloride according to the invention are prepared. Therefore, the above-mentioned predetermined organic solvent having an appropriate polarity is used in this reaction process. In particular, as such organic solvent, methoxyethanol, ethoxyethanol, methoxymethanol, ethoxymethanol, methanol, ethanol, propanol, isopropanol, Any or two selected from the group consisting of butanol, nitromethane, acetonitrile, methyl acetate, ethyl acetate, methyl acetoacetate, ethyl acetoacetate, tetrahydrofuran, 1,4-dioxane, methane chloride, chloroform, carbon tetrachloride and acetone It is preferable to use the above organic solvent. In addition, among these organic solvents, ethoxyethanol, ethanol, 2-propanol, acetonitrile, ethyl acetate, chloroform, carbon tetrachloride or acetone can be more preferably used. By using such an organic solvent, the specific crystalline form of the cytagliptin hydrochloride according to the present invention can be produced with higher purity and higher yield.

As described above, according to the preparation method of the present invention, a) a hydrochloride salt of an amine compound having a predetermined pKa or a mixture of an amine compound and hydrochloric acid and b) a cytagliptin is reacted to the present invention. Certain crystalline forms of cytagliptin hydrochloride according to the invention can be prepared. Therefore, in order to prepare crystalline Form IV of cytagliptin phosphate anhydride disclosed in WO 05/030127, there is no problem such as having to go through a complicated two-step manufacturing process or accompanied by a lower yield or purity of the active ingredient. Do not.

On the other hand, the present invention also includes a pharmaceutically effective amount of cytagliptin hydrochloride and a pharmaceutically acceptable carrier, wherein the cytagliptin hydrochloride consists of a specific crystalline form of cytagliptin hydrochloride according to the present invention which is substantially pure. It provides a pharmaceutical composition. For example, the cytagliptin hydrochloride contained in the pharmaceutical composition of the present invention contains a specific crystalline form of the cytagliptin hydrochloride according to the present invention in a crystal purity of 90% or more.

Such a pharmaceutical composition contains, as an active ingredient, a specific crystalline form of the cytagliptin hydrochloride according to the present invention which shows little hygroscopicity and shows high stability, thereby minimizing decomposition of the active ingredient by moisture, thereby ensuring high purity of the active ingredient for a long time. It can be stored in a state that maintains high efficacy against type 2 diabetes for a long time.

Such a pharmaceutical composition according to the present invention exhibits a high inhibitory activity against DP-IV for a long time, and thus may be very useful as a pharmaceutical composition for treating or preventing type 2 diabetes.

On the other hand, the pharmaceutical composition according to the present invention can be administered through any route of administration, such as oral, parenteral, inhalation spray, nasal, vaginal, rectal, sublingual, topical, and the like according to the route of administration Can be prepared in a suitable dosage unit dosage form containing an acceptable carrier, adjuvant or vehicle. Since the configuration and preparation method of each unit dosage form is according to the configuration obvious to those skilled in the art according to the route of administration and the type of unit dosage form, a detailed description thereof will be omitted.

In addition, the pharmaceutical composition of the present invention may be prepared in a dosage form suitable for oral administration such as tablets, troches, lozenges, aqueous or oily suspensions, dispersion powders or granules, emulsions, hard or soft capsules, syrups or elixirs. Can be. In this case, the pharmaceutical composition of the present invention may be prepared according to any method known in the art for preparing oral pharmaceutical compositions.

In addition, the oral pharmaceutical composition of the present invention may include a suitable pharmaceutically acceptable non-toxic excipient according to the form of each formulation. For example, the excipient may be an inert diluent such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; Granules or disintegrants such as cornstarch, or alginic acid; Binders such as starch, gelatin or acacia; Lubricants such as magnesium stearate, stearic acid or talc.

In addition, any of the pharmaceutically acceptable excipients which were obviously used by those skilled in the art according to the form of each formulation may be used in the oral pharmaceutical composition of the present invention, and the preparation method thereof is also obvious to those skilled in the art according to the form of each formulation. According to one manufacturing method.

Hereinafter, the configuration and operation of the present invention through the preferred embodiment of the present invention will be described in more detail. However, the scope of the present invention is not determined according to these embodiments, which are merely presented as examples.

Comparative Example 1 Preparation of Citagliptin Hydrochloride Disclosed in the Prior Art (WO 03/004498) (See Example 7 of WO 03/004498)

According to the method described in step A of Example 7 of WO 03/004498, (3R) -3- [(1, 1-dimethylethoxy-carbonyl) -amino] -4- (2, 4, 5-tri Fluorophenyl) -butanoic acid and 3- (trifluoromethyl) -5, 6, 7, 8-tetrahydro-1, 2, 4-triazolo [4, 3-α] pyrazine by reacting 7- [ (3R) -3-[(1,1-dimethylethoxycarbonyl) amino] -4- (2,4,5-trifluorophenyl) butanoyl] -3- (trifluoromethyl) -5, 6,7,8-tetrahydro-1,2,4-triazole- [4,3-a] pyrazine was prepared.

¹ H NMR (500 MHz, CDCl ₃ ) δ 1.37 (s, 9H), 2.61-3.00 (m, 4H), 3.92-4.30 (m, 5H), 4.93 (s, 1H), 4.95-5.12 (m, 1H) , 5.22-5.35 (br, 1 H), 6.83-6.95 (m, 1 H), 7.02-7.12 (m, 1 H). LC / MS (M + 1-t-Bu) 452.

2 ml of a hydrochloric acid solution saturated in methanol was added to 7-[(3R) -3-[(1,1-dimethylethoxycarbonyl) amino] -4- (2,4,5-trifluorophenyl) butanoyl] After adding dropwise to 22 mg (0.039 mmol) of -3- (trifluoromethyl) -5,6,7,8-tetrahydro-1,2,4-triazole- [4,3-a] pyrazine, The mixture was stirred for 1 hour at, and the solvent was depressurized to obtain 16.5 mg of the title cytagliptin hydrochloride.

¹ H NMR (500 MHz, CD ₃ OD) δ 2.75-3.15 (m, 4H), 3.82-4.35 (m, 5H), 4.90-5.05 (m, 2H), 7.16-7.25 (m, 1H), 7.30-7.42 (m, 1 H). LC / MS (M + 1) 408.

Comparative Example 2: Preparation of Form IV of Cytagliptin Phosphate Anhydride disclosed in the prior art (WO 05/030127) (see Example of WO 05/030127)

First, the cytagliptin base was prepared from the cytagliptin hydrochloride prepared in Comparative Example 1.

Thereafter, 15.0 g (36.9 mmol) of cytagliptin base, 31.5 ml of isopropanol, 13.5 ml of water, and 4.25 g (36.9 mmol) of 85% aqueous phosphoric acid solution were added to a 250 ml round bottom flask and mixed. This mixture was heated to 75 ° C. A white precipitate was formed at low temperatures but it was found to dissolve up to 75 ° C. The solution was cooled to 68 ° C. and held at this temperature for 2 hours. During maintenance, a solid slurry bed formed. The temperature of this slurry was cooled to 21 ° C. at 4 ° C. per hour and then maintained overnight. Subsequently, 105 ml of isopropanol was added to the slurry. After 1 hour, the slurry was filtered and washed with 45 ml of isopropanol. The solid formed was dried on frit while exposed to air. 18.6 g of cytagliptin phosphate monohydrate was obtained in solid form. It was confirmed that the cytagliptin phosphate monohydrate obtained by HPLC was 99.8% or more.

Form IV of cytagliptin phosphate anhydride was prepared by heating the cytagliptin phosphate monohydrate at 120 ° C. for about 2 hours or at about 58 ° C. or higher for about 8 hours.

Example 1 Preparation of Certain Crystalline Forms of Cytagliptin Hydrochloride According to the Present Invention

First, the cytagliptin base was prepared from the cytagliptin hydrochloride prepared in Comparative Example 1.

After dissolving 2 g (5 mmol) of the prepared cytagliptin base in 20 ml of methane chloride solvent, a solution of 1.9 g (110 mmol) of trialinamine hydrochloride in 4 ml of methane chloride was added dropwise at room temperature. At this time, the dropping process and the reaction process were performed in a state in which water in the air is blocked. Thereafter, the mixed solution in which the reaction was completed was crystallized by stirring. The crystals were filtered to obtain 2.1 g (yield: 95%) of the crystalline form of cytagliptin hydrochloride according to the present invention.

¹ H NMR (MeOH- d ₄ , 300 MHz) δ 7.37-7.36 (m, 1H), 7.24-7.19 (m, 1H), 5.01-4.92 (m, 2H), 4.30-4.22 (m, 2H), 4.01-3.88 (m, 3H), 3.07-3.03 (m , 2H), 2.95-2.83 (m, 2H)

Example 2 Preparation of Certain Crystalline Forms of Cytagliptin Hydrochloride According to the Present Invention

First, the cytagliptin base was prepared from the cytagliptin hydrochloride prepared in Comparative Example 1.

After dissolving 306 mg (0.75 mmol) of the prepared cytagliptin base in 3 ml of acetonitrile solvent, 104 mg (0.90 mmol) of pyridine hydrochloride were added and stirred at room temperature for 12 hours. After completion of the reaction, the reaction solution was completely removed, and about 5 ml of methane chloride was added thereto, followed by stirring for 30 minutes. The white solid produced was filtered to give 295 mg (yield 88.5%) of the crystalline form of cytagliptin hydrochloride according to the present invention.

¹ H NMR (MeOH- d ₄ , 300 MHz) δ 7.37-7.36 (m, 1H), 7.24-7.19 (m, 1H), 5.01-4.92 (m, 2H), 4.30-4.22 (m, 2H), 4.01-3.88 (m, 3H), 3.07-3.03 (m , 2H), 2.95-2.83 (m, 2H)

Example 3 Preparation of Certain Crystalline Forms of Cytagliptin Hydrochloride According to the Present Invention

First, the cytagliptin base was prepared from the cytagliptin hydrochloride prepared in Comparative Example 1.

312 mg (0.76 mmol) of the prepared cytagliptin base was dissolved in 3 ml of methane chloride solvent, and then 106 mg (0.92 mmol) of pyridine hydrochloride were added thereto, followed by stirring at room temperature for 12 hours. The white solid produced was filtered to obtain 343 mg (yield 99%) of the crystalline form of cytagliptin hydrochloride according to the present invention.

¹ H NMR (MeOH- d ₄ , 300 MHz) δ 7.37-7.36 (m, 1H), 7.24-7.19 (m, 1H), 5.01- 4.92 (m, 2H), 4.30-4.22 (m, 2H), 4.01-3.88 (m, 3H), 3.07-3.03 (m , 2H), 2.95-2.83 (m, 2H)

Example 4 Preparation of Certain Crystalline Forms of Cytagliptin Hydrochloride According to the Present Invention

First, the cytagliptin base was prepared from the cytagliptin hydrochloride prepared in Comparative Example 1.

310 mg (0.76 mmol) of the prepared cytagliptin base was dissolved in 3 ml of a methane chloride solvent, and then 159 mg (0.91 mmol) of triallylamine hydrochloride was added thereto, followed by stirring at room temperature for 12 hours. The white solid produced was filtered to obtain 319 mg (yield 93.2%) of the crystalline form of cytagliptin hydrochloride according to the present invention.

¹ H NMR (MeOH- d ₄ , 300 MHz) δ 7.37-7.36 (m, 1H), 7.24-7.19 (m, 1H), 5.01-4.92 (m, 2H), 4.30-4.22 (m, 2H), 4.01-3.88 (m, 3H), 3.07-3.03 (m , 2H), 2.95-2.83 (m, 2H)

Test Example 1 Confirmation of Crystal Form of Cytagliptin Hydrochloride

Requested by the Korea Institute of Chemistry, DSC thermal analysis, powder X-ray diffraction pattern and IR spectrum of each cytagliptin hydrochloride prepared in Examples 1 to 4 and Comparative Example 1 were measured, and the results are shown in FIG. 1 to 6 are shown. More specifically, DSC thermal analysis (FIG. 2), powder X-ray diffraction pattern (FIG. 1) and IR spectrum (FIG. 3) of specific crystal forms of the cytagliptin hydrochloride prepared in Examples 1 to 4 are shown in FIG. 3 to 3, DSC thermal analysis (FIG. 5), powder X-ray diffraction pattern (FIG. 4) and IR spectrum (FIG. 6) of the cytagliptin hydrochloride prepared in Comparative Example 1 are shown in FIGS. 6 is shown.

For reference, in order to measure the DSC thermal analysis, powder X-ray diffraction pattern and IR spectrum, respectively, TA instrument 2910 DSC (DSC thermal analysis device), Bruker D / 8 Discover X-ray Diffractometer with GADDS (powder X-ray diffraction) Analysis device) and SensIR Technology TravelIR II (IR analysis device).

As a result of this measurement, in the specific crystal form of the cytagliptin hydrochloride of Examples 1 to 4, that is, the powder X-ray diffraction pattern (Fig. 1) of the specific crystal form of the cytagliptin hydrochloride according to the present invention, basically 13.7, 18.0, Significantly strong characteristic peaks appear at 2θ at 22.6, 25.7 and 27.0 ° ± 0.2 °, whereas the cytagliptin hydrochloride of Comparative Example 1, ie, the powder of cytagliptin hydrochloride according to the prior art (WO 03/004498) It was confirmed that these characteristic peaks did not appear at all in the X-ray diffraction pattern (FIG. 4).

In addition, DSC thermal analysis of the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4 (Fig. 2) showed a clear endothermic peak at 135.08 ℃, DSC thermal analysis of the cytagliptin hydrochloride of Comparative Example 1 5, only the exothermic peak appeared at 123.33 degreeC.

In addition, it was confirmed that the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4 (FIG. 3) is distinguished from the cytagliptin hydrochloride (FIG. 6) of Comparative Example 1 also in the IR spectrum.

Based on the above measurement results, the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4, that is, the specific crystalline form of the cytagliptin hydrochloride according to the present invention, is the cytagliptin hydrochloride of Comparative Example 1, namely, It was confirmed that the crystal form was completely different from cytagliptin hydrochloride according to the technique (WO 03/004498).

In addition, the measurement results of FIGS. 1 to 3 were compared with DSC thermal analysis and powder X-ray diffraction pattern (see FIGS. 1 to 5 of WO 05/030127) for Form IV of cytagliptin phosphate anhydride of Comparative Example 2. As a result of comparison, the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4, that is, the specific crystalline form of the cytagliptin hydrochloride according to the present invention is different from the crystalline form IV of the cytagliptin phosphate anhydride of Comparative Example 2 The form was confirmed.

Test Example 2: DP-IV enzyme inhibitory activity test

The DP-IV enzyme inhibitory activity of the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4, the crystalline form IV of the cytagliptin hydrochloride of Comparative Example 1 and the cytagliptin phosphate anhydride of Comparative Example 2 was determined. Comparative test.

In order to compare the enzyme inhibitory activity, a test method of basically dissolving each compound in the appropriate concentration and reacting with the enzyme for each concentration was applied.

More specifically, the reaction solution was used 50mM HEPES (pH 8.0), Ala-Pro-AFC was used as the substrate. First, 10ul of each compound, 20ul of enzyme and 145ul of the reaction solution were added to react at room temperature for 30 minutes, and the substrate was added thereto, followed by enzymatic reaction at 37 ° C for 30 minutes.

After the reaction was completed, the fluorescence value was measured under Excitation 400nm and Emission 508nm using a fluorescence photometer and the activity inhibition was calculated by comparing with the control.

The inhibitory activity against the DP-IV enzyme of each compound measured in this manner is shown in Table 1 below.

TABLE 1

Examples 1-4 Comparative Example 1 Comparative Example 2 Enzyme Inhibitory Effects (IC ₅₀ ) 44 nM 45 nM 45 nM

Referring to Table 1 above, the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4, ie, the specific crystalline form of the cytagliptin hydrochloride according to the present invention, is citagliptin according to the prior art (WO 03/004498). It has been confirmed that hydrochloride and DP-IV inhibitory activity equivalent to crystalline Form IV of cytagliptin phosphate anhydride disclosed in WO 05/030127 show a pharmacological activity useful for treating or preventing type 2 diabetes.

Test Example 3 Hygroscopicity Comparison

Hygroscopicity in air was measured for the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4, the crystalline form IV of the cytagliptin hydrochloride of Comparative Example 1 and the cytagliptin phosphate anhydride of Comparative Example 2 above.

The conditions of the hygroscopic measurement are as follows, and the measurement results are summarized in Table 2 below.

Hygroscopicity measurement conditions

Temperature and humidity conditions: 25 ℃, 45% humidity

Exposure time in air: 24 hours

Measurement method: Karl-fisher measurement titration

TABLE 2

Examples 1-4 Comparative Example 1 Comparative Example 2 Moisture content before air exposure 0.98% 1.02% 1.85% Moisture content after 24 hours exposure 1.35% 8.8% 4.62%

Referring to Table 2, the cytagliptin hydrochloride of Comparative Example 1 absorbs about 7.8% of water in air for 24 hours, and Form IV of cytagliptin phosphate anhydride of Comparative Example 2 is also about 2.8% of water. While the specific crystalline form of the cytagliptin hydrochloride of Examples 1-4 absorbed about 0.37% moisture.

That is, the specific crystalline form of the cytagliptin hydrochloride of Examples 1 to 4, that is, the specific crystalline form of the cytagliptin hydrochloride according to the present invention is about 1 / of the cytagliptin hydrochloride according to the prior art (WO 03/004498). 20, and also exhibits extremely low hygroscopicity, which is only about 1 / 7.5 of crystalline Form IV of cytagliptin phosphate anhydride disclosed in WO 05/030127, thus exhibiting a markedly high stability in air.

As described above, according to the present invention, it is possible to provide a novel crystalline form of cytagliptin hydrochloride which has minimal hygroscopicity and markedly improved water stability compared to the pharmaceutical salt of cytagliptin or a crystalline form thereof used in the prior art.

Therefore, the novel crystal form of the cytagliptin hydrochloride can be used to provide a pharmaceutical composition and a medicament for treating type 2 diabetes, which have very high purity of the active ingredient, high stability over time, and which can maintain medicinal effects for a long time.

Moreover, the novel crystalline form of the hydrochloride salt of cytagliptin has the advantage that it can be produced in high yield through a very simple method.

Claims (10)

Crystalline form of cytagliptin hydrochloride having a powder X-ray diffraction pattern showing a peak at 2θ at 13.7, 18.0, 22.6, 25.7, and 27.0 ° ± 0.2 °. The powder of claim 1, wherein the powder exhibits peaks at 2θ of 6.5, 8.0, 13.7, 16.0, 18.0, 18.7, 19.6, 19.8, 20.3, 22.6, 23.4, 24.8, 25.4, 25.7, 27.0, 29.4, 32.3 ° ± 0.2 ° Crystalline Form Of Cytagliptin Hydrochloride With X-ray Diffraction Patterns. 6.3, 6.5, 7.6, 8.0, 13.2, 13.7, 16.0, 18.0, 18.7, 19.6, 19.8, 20.3, 21.7, 22.6, 23.4, 23.8, 24.8, 25.4, 25.7, 26.5, 27.0, 27.6, Powder X-ray diffraction pattern showing peaks at 2θ of 28.8, 29.4, 29.8, 31.0, 31.6, 32.0, 32.3, 32.9, 33.3, 34.2, 34.8, 35.6, 36.0, 36.5, 37.5, 38.3, 38.7, 43.3 ° ± 0.2 ° Crystalline form of cytagliptin hydrochloride. 4. The crystalline form of cytagliptin hydrochloride according to claim 3, having a powder X-ray diffraction pattern shown in FIG. Under any one or two or more organic solvents selected from the group consisting of hydroxy solvents, nitroalkane solvents, acetonitrile solvents, acetate solvents, ether solvents, alkane chloride solvents and ketone solvents, a) The citagliptin according to any one of claims 1 to 4, comprising reacting a hydrochloride or mixture of an amine compound and hydrochloric acid of an amine compound having a pKa less than the maximum pKa, and b) cytagliptin. Method for preparing the crystalline form of the hydrochloride. The method of claim 5, further comprising the step of promoting the crystallization of the hydrochloride salt of cytagliptin by stirring the reaction product. The hydrochloride or amine-based compound and a mixture of hydrochloric acid of an amine compound having a pKa less than the maximum pKa of cytagliptin are a hydrochloride salt of a trialinamine, a hydrochloride salt of pyridine, a triallinamine salt and a hydrochloric acid. A process for preparing the crystal form of cytagliptin hydrochloride which is a mixture of or a mixture of pyridine and hydrochloric acid. The method of claim 5, wherein the organic solvent is methoxyethanol, ethoxyethanol, methoxymethanol, ethoxymethanol, methanol, ethanol, propanol, isopropanol, butanol, nitromethane, acetonitrile, methyl acetate, ethyl acetate, methyl aceto A method for producing a crystalline form of cytagliptin hydrochloride, which is one or two or more organic solvents selected from the group consisting of acetate, ethyl acetoacetate, tetrahydrofuran, 1,4-dioxane, methane chloride, chloroform, carbon tetrachloride and acetone. A pharmaceutically effective amount of cytagliptin hydrochloride and a pharmaceutically acceptable carrier, wherein the cytagliptin hydrochloride has a crystalline purity of at least 90% of the crystal form of cytagliptin hydrochloride according to any one of claims 1 to 4. Pharmaceutical composition comprising a. The pharmaceutical composition of claim 9, which is used for the treatment or prevention of type 2 diabetes.

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