HK1114849A - Crystalline forms of a known pyrrolidine factor xa inhibitor - Google Patents

Description

Crystalline forms of a known pyrrolidine factor Xa inhibitor

Technical Field

The present invention relates to crystalline forms of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1), which exhibit inhibitory effect on the serine protease factor Xa. In particular, the present invention relates to 1, 2-pyrrolidinedicarboxamide, crystalline forms A, B and C of N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) and methods of using them as therapeutic agents for treating diseases characterized by abnormal thrombosis in a mammal.

Background

Ischemic heart disease and cerebrovascular disease are the leading causes of death in the world. Abnormal coagulation and inappropriate thrombosis within blood vessels contributes to many acute cardiovascular diseases.

Thrombin can be considered a key or major regulatory enzyme in the coagulation cascade; as a positive and negative feedback regulator in normal hemostasis, it has multiple effects. However, in certain pathological conditions, positive feedback regulation is amplified by the catalytic activation of cofactors required for thrombin generation. Such cofactors include factor Xa, a serine protease, which occupies a critical position in the coagulation cascade.

Abnormal intravascular coagulation and inappropriate thrombosis contribute to a number of cardiovascular diseases such as myocardial infarction, myocardial ischemia, stroke associated with atrial fibrillation, Deep Vein Thrombosis (DVT), pulmonary embolism, cerebral ischemia or infarction, peripheral arterial disease, restenosis, atherosclerosis, and thromboembolism. In addition, thrombosis is also associated with non-cardiovascular diseases, such as cancer, diabetes and sepsis. Currently, some of these conditions are treated with antithrombotic agents. However, many such agents require close monitoring of the patient to prevent bleeding. It has recently been recognized that inhibition of factor Xa provides sustained antithrombotic protection. In animal studies, short-term exposure to factor Xa inhibitors can produce a sustained antithrombotic effect. The data show that factor Xa inhibition potentially provides a large therapeutic window between antithrombotic effect and bleeding tendency. Thus, there is a range in which inhibition of factor Xa can be achieved without simultaneously increasing the sensitivity of the patient to bleeding, unlike currently available drugs.

Sepsis is a complex extension of acute inflammation and involves a progressive amplification cycle of coagulation and inflammation. The close involvement of the coagulation system in the progression of this disease has led to treatments that include antithrombotic agents. However, the currently available antithrombotic agents do not provide an adequate treatment for the disease.

There is a well-known association between malignancy and thrombosis. Recent evidence shows that factor Xa plays a role in tumor metastasis independently of its role in thrombosis and hemostasis.

Type 2 diabetic patients who have not previously had clinical coronary artery disease may have the same likelihood of death from coronary artery disease as non-diabetic patients who have previously had a myocardial infarction. Increased cardiovascular risk in diabetes is contributed by the accumulation of cardiovascular risk factors, including hypertension, dyslipidemia, hyperinsulinemia, hyperglycemia, obesity, and hemostatic risk factors such as hyper-fibrinogen and increased levels of plasminogen activator inhibitor-1. These risk factors combine to produce life-threatening thrombotic conditions that can be effectively reduced by treatment with a factor Xa inhibitor.

Factor Xa inhibitors are known in the art and one of the compounds, Ximelgatran, has recently been approved for marketing in europe. However, it is apparent that there remains a need for more effective agents that can modulate factor Xa proteolytic activity.

Various methods for preparing cyclic amino acids and proline derivatives, all of which inhibit factor Xa, are described in U.S. patent application No. US2003/0162787A1 (the' 787 application) to Bigge et al. Example 150 more specifically describes the synthesis of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). (see (2R, 4R) 4-methoxy-pyrrolidine-1, 2-dicarboxylic acid 1- [ (4-chlorophenyl) -amide ]2- { [ 2-fluoro-4- (2-oxo-2H-pyridin-1-yl) -phenyl ] -amide } in the' 787 application).

Chemical and physical properties are important in the commercial development of pharmaceutical compounds. These properties include, but are not limited to: (1) loading properties such as molar volume, density and hygroscopicity, (2) thermodynamic properties such as melting temperature, vapor pressure and solubility, (3) kinetic properties such as dissolution rate and stability (including stability under ambient conditions, especially stability to humidity and under storage conditions), (4) surface properties such as surface area, wettability, interfacial tension and shape, (5) mechanical properties such as hardness, tensile strength, compactibility, handleability, flowability and blendability, (6) filtration properties and (7) bioavailability. These properties can influence, for example, the processing and storage of compositions comprising 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1- (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).

To improve these chemical and physical properties, crystalline forms of N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) that provide improvements in one or more of these properties over the amorphous forms of 1, 2-pyrrolidine dicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) are desirable.

During the development of a drug, it is generally envisaged that it is important to find the most stable crystalline form of the drug. This most stable crystalline form is the form that is likely to have the best chemical stability and thus the longest shelf life in the formulation. However, it is also advantageous to have multiple drug forms, such as salts, hydrates, crystalline forms and non-crystalline forms. Because different physical forms offer different advantages, there is no perfect physical form of a drug. Finding the most stable form, and such other forms, is laborious and the results are unpredictable.

We have surprisingly and unexpectedly found crystalline forms of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1), designated forms A, B and C.

Brief description of the invention

Thus, the present invention includes crystalline forms of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).

The chemical formula of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) is as follows:

one embodiment of the present invention is crystalline form a (form a) of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). Form a is characterized by an X-ray powder diffraction (PXRD) pattern (table 1) and/or a Nuclear Magnetic Resonance (NMR) spectrum (table 4).

Another embodiment of the invention is crystalline form B (form B) of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). Form B is characterized by an X-ray powder diffraction (PXRD) pattern (table 2) and/or a Nuclear Magnetic Resonance (NMR) spectrum (table 4).

Another embodiment of the invention is crystalline form C of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) (form C). Form C is characterized by an X-ray powder diffraction (PXRD) pattern (table 3) and/or a Nuclear Magnetic Resonance (NMR) spectrum (table 4).

Other embodiments of the invention include, but are not limited to: a crystalline form having a powder X-ray diffraction pattern comprising at least one peak at 6.0, 16.1, 19.7, 23.2, or 25.4 degrees 2 Θ;

a crystalline form having a powder X-ray diffraction pattern comprising peaks at 19.7 and 23.2 degrees 2 Θ and one or more other peaks at 16.1 or 21.9 degrees 2 Θ;

a crystalline form having a powder X-ray diffraction pattern with peaks at 19.7 and 23.2 degrees 2 Θ and having one or more solid state NMR chemical shifts at 173.8 or 111.3 ppm;

a crystalline form having a powder X-ray diffraction pattern with a spectral peak at 16.1, 19.7, or 21.9 degrees 2 Θ and having one or more solid state NMR chemical shifts at 173.8 or 111.3 ppm;

a crystalline form having a powder X-ray diffraction pattern comprising at least one peak at 18.9, 25.9, 26.0, 28.7, or 34.8 degrees 2 Θ;

a crystalline form having a powder X-ray diffraction pattern comprising peaks at 26.0 and 25.9 degrees 2 Θ and one or more other peaks at 18.9 or 21.8 degrees 2 Θ;

a crystalline form having a powder X-ray diffraction pattern with a spectral peak at 25.9 or 26.0 degrees 2 Θ and having one or more solid state NMR chemical shifts at 172.9 or 110.0 ppm;

a crystalline form having a powder X-ray diffraction pattern comprising at least one peak at 18.9 or 21.8 degrees 2 Θ and having one or more solid state NMR chemical shifts at 172.9 or 110.0 ppm;

a crystalline form having a powder X-ray diffraction pattern with at least one spectral peak at 13.5 or 17.6 degrees 2 Θ;

a crystalline form having a powder X-ray diffraction pattern comprising peaks at 13.5 and 17.6 degrees 2 Θ and one or more other peaks at 9.2, 18.3, or 22.5 degrees 2 Θ;

a crystalline form having a powder X-ray diffraction pattern with a spectral peak at 13.5 or 17.6 degrees 2 Θ and having one or more solid state NMR chemical shifts at 174.3, 105.4, or 130.3 ppm;

a crystalline form having a powder X-ray diffraction pattern with at least one peak at 9.2, 13.5, 17.6, 18.3, or 22.5 degrees 2 Θ and having one or more solid state NMR chemical shifts at 174.3, 105.4, or 130.3 ppm.

Another embodiment of the invention is a composition comprising one or more of the above forms and a pharmaceutically acceptable excipient, diluent or carrier.

Another embodiment of the invention is a composition comprising one or more of the above forms and a pharmaceutically acceptable excipient, diluent or carrier and one or more of the following agents: a non-steroidal anti-inflammatory drug, a thrombin inhibitor, a factor VIIa inhibitor, a platelet aggregation inhibitor, a vitamin K antagonist, a GPIIbIIIa antagonist, a heparinoid, a thrombolytic agent, or a fibrinolytic agent.

A more specific embodiment of the present invention is the above composition, wherein the non-steroidal anti-inflammatory drug is one of: aspirin, ibuprofen, naproxen sodium (naproxensodium), indomethacin, celecoxib, valdecoxib or piroxica. The thrombin inhibitor is one of the following: agatroban, effegatran, inogatran (inogatran), hirudin, hirulog, ximelagatran or melagatran. The platelet aggregation inhibitor is one of the following: dipyrimidole, agrrenox, clopidogrel, ticlopidine or P2Y12 inhibitors. The vitamin K antagonist is one of the following: warfarin (coumadin), warfarin (warfarin), or a coumarin derivative. The GPIIbIIIa antagonist is one of the following: abciximab, epifibatide or tirofiban. The heparinoids are: heparin, fast-avoiding (fraxiparin), tinzaparin (tinzaparin), idraparanux, dermatan sulfate, fondaparinux or enoxaparin. The thrombolytic or fibrinolytic agent is one of: tissue plasminogen activators, urokinase, streptokinase, plasminogen activator inhibitor-1 inhibitors, or inhibitors of thrombin-activatable fibrinolysis inhibitors.

The crystalline forms of the invention or mixtures of said forms can be administered in a therapeutically effective amount to a mammal in which the use of a factor Xa inhibitor is indicated. Mammals as used herein include, but are not limited to, humans.

Other embodiments of the invention include, but are not limited to: a method of treating acute, subacute or chronic thrombotic disorders in a mammal with a therapeutically effective amount of a crystalline form or composition of the invention.

A method of treating primary deep vein thrombosis or secondary deep vein thrombosis in a mammal with a therapeutically effective amount of a crystalline form or composition of the invention. A method of treating atrial fibrillation in a mammal with a therapeutically effective amount of a crystalline form or composition of the invention.

A method of treating the following conditions in a mammal with a therapeutically effective amount of a crystalline form or composition of the invention: venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, cerebral infarction, restenosis, atherosclerosis, angina pectoris, primary deep vein thrombosis, secondary deep vein thrombosis, cancer, sepsis, diabetes or thromboembolism associated with cardiovascular disease.

Other embodiments of the invention include, but are not limited to: use of at least one form a, form B or form C in the manufacture of a medicament; use of a crystalline form or composition of the invention in the manufacture of a medicament for the treatment of a condition in a mammal which condition is beneficially treated by inhibition of factor Xa; use of a crystalline form or composition of the invention in the manufacture of a medicament for the treatment of acute, subacute or chronic thrombotic disorders; use of a crystalline form or composition of the invention in the manufacture of a medicament for the treatment of primary deep vein thrombosis or secondary deep vein thrombosis; use of a crystalline form or composition of the invention in the manufacture of a medicament for the treatment of a thromboembolic event in a mammal having atrial fibrillation; use of a crystalline form or composition of the invention in the manufacture of a medicament for the treatment of: use of venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, cerebral infarction, restenosis, atherosclerosis, angina pectoris, primary deep vein thrombosis, secondary deep vein thrombosis, cancer, diabetes, or thromboembolism associated with a cardiovascular disease, or a crystalline form of the invention, in the manufacture of a medicament for the treatment of: venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, cerebral infarction, restenosis, atherosclerosis, angina pectoris, primary and secondary deep vein thrombosis, cancer, sepsis, diabetes, or thromboembolism associated with cardiovascular disease (including but not limited to acute coronary syndrome, atrial fibrillation, heart valve replacement, and deep vein thrombosis).

The crystalline forms and compositions of the present invention, or mixtures thereof, may be administered in unit dosage form contained in a package or kit. The kit includes the unit dosage form and a container. Typically, the kit includes instructions for administration of the unit dosage form according to a treatment regimen. The instructions may include instructions suggesting how to use the kit for the treatment of acute, subacute, or chronic thrombotic disorders, including, but not limited to, treatment of venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, cerebral infarction, restenosis, atherosclerosis, angina, primary and secondary deep vein thrombosis, thromboembolism associated with cardiovascular disease (including, but not limited to, acute coronary syndrome, atrial fibrillation, heart valve replacement, and deep vein thrombosis), or treatment of cancer, sepsis, and diabetes. The container may be of any conventional shape or form known in the art, for example a carton, glass or plastic bottle, or blister pack of individual dosage forms which can be pressed from the back.

Brief description of the drawings

FIG. 1A

A diffractogram of crystalline form a of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).

FIG. 1B

A diffractogram of crystalline form B of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).

FIG. 1C

Diffraction pattern of crystalline form C of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).

FIG. 2A

Solid state of form A¹³C nuclear magnetic resonance spectroscopy.

FIG. 2B

Solid state of form B¹³C nuclear magnetic resonance spectroscopy.

FIG. 2C

Solid state of form C¹³C nuclear magnetic resonance spectroscopy.

FIG. 3 of the drawings

Differential Scanning Calorimetry (DSC) patterns of forms A, B and C.

Detailed description of the invention

Definition of

1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) is also known as (2R, 4R) 4-methoxy-pyrrolidine-1, 2-dicarboxylic acid 1- [ (4-chlorophenyl) -amide ]2- ([ 2-fluoro-4- (2-oxo-2H-pyridin-1-yl) -phenyl ] -amide, depending on the nomenclature used to identify the compound.

The term "form a, form B and form C" as used herein refers to the crystalline forms of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). The references to "form a", "form a polymorph", "crystalline form a" and "form a polymorph of 1, 2-pyrrolidine dicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1)" are the same and are used interchangeably herein. The references to "form B", "form B polymorph", "crystalline form B", and "form B polymorph of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1)" are the same and are used interchangeably herein. The references to "form C", "form C polymorph", "crystalline form C" and "form C polymorph of 1, 2-pyrrolidine dicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1)" are the same and are used interchangeably herein.

The terms "polymorph" and "crystalline form" are used interchangeably herein.

The terms "polymorphic form" and "polymorph" are used interchangeably herein.

The term "amorphous" when applied to 1, 2-pyrrolidinedicarboxamide, N1- (4 chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) refers to a solid state in which the 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) molecules are in a disordered arrangement and do not form a distinguishable crystal lattice or unit cell.

The term "crystalline form", "polymorphic form" or "polymorph" when applied to 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) refers to a solid state form, wherein the molecular arrangement of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) forms a distinguishable crystal lattice that produces characteristic diffraction peaks when exposed to X-ray radiation.

The term "DSC" means differential scanning calorimetry.

The term "mammal" as used herein includes, but is not limited to, humans.

The term "pharmaceutically acceptable" means suitable for use in mammals.

The term "PXRD" means powder X-ray diffraction.

The term "slurry" means a stirred suspension of a solid compound in a solvent, wherein the concentration of the compound is higher than its solubility in the solvent. "slurrying" refers to preparing a slurry.

The terms "pattern" and "diffraction pattern" are used herein in the same sense when used in conjunction with PXRD.

The terms "treatment" and "treating" as used herein include palliative, curative and prophylactic treatment.

Powder X-ray diffraction (PXRD)

Compounds having the same chemical structure may exist in different physical forms. They may be amorphous or present in different crystalline forms. Different crystalline forms often have different physical properties (i.e., bioavailability, solubility, melting point, etc.). These different crystalline forms are sometimes referred to as polymorphs. One method of determining the structure of a crystalline form is known as powder X-ray diffraction (PXRD) analysis. PXRD analysis involves the collection of crystallographic data from a set of crystals. For PXRD analysis, a powder sample of the crystalline material was placed in a holder and then placed in a diffractometer. The X-ray beam is directed at the sample, initially at a small angle relative to the plane of the support, and then moved in an arc, continuously increasing the angle between the incident beam and the plane of the support. The intensity of the reflected radiation is recorded. These data may be represented graphically in a PXRD pattern.

The differences in measurements associated with such X-ray powder analysis come from a variety of factors including: (a) errors in sample preparation (e.g., sample height), (b) instrument errors (e.g., flat sample errors), (c) calibration errors, (d) operator errors (including those in determining spectral peak positions), (e) material properties (e.g., preferred orientation and transparency errors), (f) batch-to-batch differences in compounds, and (g) machine type. Calibration errors, sample height errors, lot-to-lot variations, and machine type difference anomalies result in shifts of all spectral peaks in the same direction. These shifts can be identified from the powder X-ray diffraction pattern and can be eliminated by compensation for the shift (applying a systematic correction factor to all spectral peak position values) or recalibration of the instrument. Such correction factors are typically in the range of 0 to 0.2 degrees 2 θ.

Form a, form B and form C of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) are characterized by their PXRD pattern. The samples were placed in an aluminum holder ready for analysis. Powder X-ray diffraction (PXRD) patterns shown in fig. 1A, 1B and 1C were collected on a Rigaku (Tokyo, Japan) Ultima-plus diffractometer using CuK α radiation operating at 40kV and 40 mA. Diffracted radiation was detected using a NaI scintillation detector. The sample was scanned from 3 degrees to 50 degrees 2 theta at 2.4 seconds per step, 0.04 or 0.02 degree 2 theta steps. Alumina standards were analyzed to check instrument calibration. Sample data were collected at room temperature. The Data was processed using Materials Data inc. jade (version 3.1).

Form A is characterized by a PXRD pattern expressed in degrees 2 theta values and relative intensities ≧ 20.0 (Table 1). Form B is characterized by a PXRD pattern expressed in degrees 2 theta values and relative intensities ≧ 19.5 (Table 2). Form C is characterized by a PXRD pattern expressed in degrees 2 theta values and relative intensities ≧ 10.0 (Table 3).

Table 1 PXRD spectral peak list for form a

Degree 20	Relative Strength (%)
		5.8	43.2
6.0	25.7
		8.1	90
14.6	43.5
		16.1	22
16.6	39.3
		17.2	100.0
17.3	60.3
		18.0	73.1
18.2	41.6
		19.7	33.4
20.2	24.3
		20.6	71.7
20.8	57
		21.7	26.9
21.9	31.1
		22.9	41.1
23.2	29.7
		25.4	21.3

Table 2 PXRD spectral peak list for form B

Degree 20	Relative Strength (%)
		5.7	36.5
8.1	61.5
		9.0	36.0
14.5	20.5
		16.6	35.0
17.2	100.0
		18.1	97.5
18.9	19.5
		20.2	44.5
20.3	29.5
		20.7	41.5
21.8	36.0
		23.6	37.0
25.0	23.5
		25.9	25.0
26.0	30.0
		28.7	24.0
34.8	20.0

Table 3 PXRD spectral peak list for form C

Degree 20	Relative Strength (%)
		9.2	19.0
13.5	20.7
		14.2	11.3
17.6	14.2
		18.3	41.0
22.0	26.9
		22.5	100.0
22.8	10.4
		23.5	13.2
23.8	14.4
		25.6	25.8
26.3	10.8
		27.7	25.5
30.2	13.0

Solid state Nuclear Magnetic Resonance (NMR)

Another method for determining the structure of a crystalline form of a compound is to use solid state NMR. Representative solid state NMR spectra of forms A, B and C are shown below in fig. 2A, 2B, and 2C. Standard acquisition and processing parameters were used. For solid state NMR, the acquisition was done on a 500MHz Varian INOVA spectrometer at 125.65MHz¹³C CP/MAS data, and the methyl resonance of hexamethylbenzene (17.3ppm) as external reference. The spectrometer was equipped with a 2.5mm chemagneticsnencil probe. 3712 data points were collected over a 46kHz scan width. The total transient of 2048-. At 140kHz¹The H-decoupling field uses variable amplitude cross polarization to acquire data. The sample was rotated at 14 kHz. Table 4 lists the temporary chemical shift assignments. These designations are made based on data acquired using intermittently decoupled solid state NMR pulse sequences and information from solution NMR chemical shifts. One skilled in the art will recognize that chemical shift positions may vary from batch to batch of compounds. Furthermore, the chemical shift position may vary depending on the instrument used for the measurement. Table 4 gives the characteristic displacement values for form a, form B and form C.

TABLE 4

Chemical shift (form A)	Chemical shift (form B)	Chemical shift (form C)
			173.8	172.9	174.3
162.6	163.4	162.4
			153.5	153.7	155.8
144.1	143.6	153.2
			139.7	139.3	138.8
135.9	137.1	130.3
			127.8	135.5	124.7
125.7	127.4	113.7
			119.9	121.7	105.4
111.3	115.4	79.6
			79.4	110.0	62.2
58.2	79.5	56.4
			53.8	61.7	50.4
37.1	58.1	38.0
				53.2
	37.2

Differential Scanning Calorimetry (DSC)

Experiments were performed using a DSC2920 instrument (TA Instruments, New castle, DE). Nitrogen was used as a purge gas for the DSC sample cell (cell) at a flow rate of 50ml/min and for the refrigeration cooling system at 110 ml/min. The calorimeter was calibrated for temperature and sample cell constant using indium (melting point 156.61 ℃, melting enthalpy 28.71J/g). A sealed aluminum pan with a pinhole was used and the sample (typically 3-5mg) was heated at a rate of 10 ℃/min. Data Analysis was performed using TA Instruments, Universal Analysis 2000 software for Windows version 3.8B. One skilled in the art will appreciate that sample purity can alter the characteristics of the data obtained by DSC. Fig. 5 shows DSC traces from forms A, B and C. Depending on the experimental conditions, recrystallization and secondary melting of form C may not be observed. Table 5 below lists the melting onset temperatures for forms A, B and C.

TABLE 5

Form(s) of	Onset of melting,. degree.C
		A	211.7
B	212.7
		C	First melting: 171.5 second melting: 207.6

Forms of the 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) described herein having equivalent PXRD diffraction patterns, regardless of the degree of water and/or solvent, are within the scope of the invention. The present invention provides one or more methods of preparing 1, 2-pyrrolidinedicarboxamide, forms A, B and C of N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1), comprising forming a solution or slurry in a solvent under conditions that yield 1, 2-pyrrolidinedicarboxamide, forms A, B or C of N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). The precise conditions used to form forms A, B and C may be determined empirically and may only give a few methods that have been found to be suitable in practice.

Pharmacology, dosage and formulation

The crystalline forms of the present invention may be administered to a patient at dosage levels ranging from 0.1 to 2,000 mg/day. In another embodiment, the crystalline form of the present invention is administered to a patient in the range of 0.01 to 700 mg/day. In another embodiment, the crystalline form of the present invention is administered to a patient at a dosage level in the range of 0.1 to 300 mg/day. In another embodiment, the crystalline form of the present invention is administered to a patient at a dosage level in the range of 0.1 to 150 mg/day. However, the particular dosage employed may vary. For example, the dosage will depend on a number of factors, including the requirements of the patient, the severity of the condition being treated, and the pharmacological activity of the crystalline form of the invention employed. Determination of the optimal dosage for a particular patient is well known to those skilled in the art.

The crystalline forms of the invention are typically administered in admixture with a suitable pharmaceutical excipient, diluent or carrier selected with regard to the intended route of administration and standard pharmaceutical practice. The pharmaceutical preparation may be in unit dosage form. In this form, the formulation is subdivided into unit dosage forms containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. In addition, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

For example, the crystalline forms of the present invention may be administered orally, buccally or sublingually in the form of tablets, capsules, multiparticulates, gels, films, ovules, elixirs, solutions or suspensions, which may contain flavouring or colouring agents, for immediate-, delayed-, modified-, sustained-, pulsed-or controlled-release applications. The crystalline forms of the present invention may also be administered in a fast-delivering or fast-dissolving dosage form or as a high-energy dispersion or as coated particles. Suitable formulations of the crystalline forms of the invention may be in coated or uncoated form, as desired.

Such solid compositions, e.g. tablets, may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate, glycerol and starch (preferably corn, potato or tapioca starch); disintegrants such as sodium starch glycolate, croscarmellose sodium and certain complex silicates; and granulation binders such as polyvinylpyrrolidone, Hydroxypropylmethylcellulose (HPMC), Hydroxypropylcellulose (HPC), sucrose, gelatin, and acacia. In addition, lubricating agents such as magnesium stearate, stearic acid, glyceryl behenate and talc may also be included.

The percentage of the composition and formulation may of course vary and may conveniently be between 2 and 60% by weight of a given dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level is obtained.

The crystalline forms of the present invention are useful for treating acute, subacute, or chronic thrombotic disorders, and more particularly, the crystalline forms of the present invention are useful for treating venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, cerebral infarction, restenosis, atherosclerosis, angina pectoris, primary and secondary deep vein thrombosis, thromboembolism associated with cardiovascular disease (including, but not limited to, acute coronary syndrome, atrial fibrillation, heart valve replacement, and deep vein thrombosis). The crystalline forms of the invention are also useful in the treatment of cancer, sepsis and diabetes.

The crystalline forms are well suited for formulation for convenient administration to a mammal for the treatment of such disorders. The crystalline forms of the present invention may be administered alone or in combination with one or more therapeutic agents. These include, for example, other anticoagulants including, but not limited to, nonsteroidal anti-inflammatory drugs including, but not limited to, aspirin, ibuprofen, naproxen sodium (naproxensodium), indomethacin, celecoxib, valdecoxib, and piroxica; thrombin inhibitors, including but not limited to argatroban, effegatran, enogatran, hirudin, hirulog, ximelagatran and melagatran; vitamin K antagonists including, but not limited to, warfarin (coumadin), warfarin (warfarin), and other coumarin derivatives; a factor VIIa inhibitor; platelet aggregation inhibitors including, but not limited to, dipyrimidole, aggrenox, clopidogrel, ticlopidine or other P2Y12 antagonists; GPIIbIIIa antagonists including, but not limited to, abciximab, epifibatide and tirofiban; heparinoids including, but not limited to, heparin, fast-setting, tinzaparin, idraparanux, dermatan sulfate, fondaparinux, enoxaparin; and thrombolytic or fibrinolytic agents such as tissue plasminogen activators, urokinase or streptokinase, inhibitors of plasminogen activator inhibitor-1, and inhibitors of thrombin-activatable fibrinolysis inhibitors. If administered in a combination of active agents, these agents may be administered simultaneously, separately or sequentially. The following non-limiting examples serve to illustrate the processes that can be used to prepare the crystalline forms of the present invention.

Examples

Example 1

(2R, 4R) -4-methoxy-pyrrolidine-2-carboxylic acid [ 2-fluoro-4- (2-oxo-pyridin-1-yl) phenyl ] -amide

Step 1 preparation of (2R, 4R) -4-methoxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

To a 500ml 3-necked flask equipped with a mechanical stirrer and a thermocouple purged with nitrogen was added 60% (w/w) NaH (8g, 200mmol) and hexane (250 ml). The mixture was stirred for 1 minute, after which the agitation was stopped and the solid was allowed to settle. The hexane was removed using a filter (candle filter). THF (250ml) and CH were then added to the flask₃I (6.51ml, 105mmol) and the resulting mixture was cooled to 0 ℃ in an ice bath. Then, (R, R) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester (22g, 95mmol) was added in portions while maintaining the reaction temperature at 5 ℃ or less. The reaction was allowed to warm to room temperature overnight. Adding H to the reaction mixture₂O (100ml), 1N HC1(100ml) and NaCl (42 g). The reaction was stirred for 10 minutes. Separating the liquid layer, and using MgSO 2 as the organic layer₄Dried, filtered and concentrated to a thick oil. When the solid just begins to precipitate, addHexane (50ml) was added and a precipitate formed immediately. The mixture was filtered to give the title compound as a white or off-white solid (20.16 g). After standing for 1 day, the filtrate was filtered to give a second crop of the title compound (1.42 g). The two batches were combined to give the title compound as a white to off-white solid (21.58g, 93% yield; chiral purity by chiral HPLC: 100%).

Step 2: 1- (4-amino-3-fluoro-phenyl) -pyridin-2-one

2-fluoro-4-iodoaniline (10.0g, 42.2mmol) was reacted with delta-valerolactam (6.27g, 63.3mmol), Cul (0.804g, 4.22mmol) and K₃P04(22.4g, 105mmol) was mixed. 1, 4-dioxane (60ml) was added followed by trans-1, 2-diaminocyclohexane (1.01ml, 8.44 mmol). After heating the mixture to reflux for 22 hours, it was cooled and diluted with EtOAc. The mixture was filtered through a plug of silica gel eluting with EtOAc, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash chromatography to give the title compound as a brown solid (3.40g, 39%). MS: APC I (AP)⁺)：209.1(M)⁺。

And step 3: (2R, 4R) -2- [ 2-fluoro-4- (2-oxo-pyridin-1-yl) -phenylcarbamoyl ] -4-methoxy-pyrrolidine-1-carboxylic acid tert-butyl ester

To the compound of step (1) (0.250g, 1.02mmol) in CHCl₃To a solution in (10ml) were added the compound of step (2) (0.212g, 1.02mmol), EEDQ (0.302g, 1.22mmol) and triethylamine (0.213ml, 1.53 mmol). After stirring to reflux the solution for 19 hours, cool to room temperature and add EtOAc. The solution was washed successively with 10% aqueous citric acid, 1N NaOH, water and brine, then MgSO₄Dried and concentrated under reduced pressure. The crude product was purified by flash chromatography to give the title compound as a brown foam (0.329g, 74%). MS: APCI (AP)⁺)：436.1(M)⁺，(AP-)：434.1(M)^-。

And 4, step 4: (2R, 4R) -4-methoxy-pyrrolidine-2-carboxylic acid [ 2-fluoro-4- (2-oxo-pyridin-1-yl) -phenyl ] -amide

To the compound of step 3 (0.329g, 0.761mmol) in anhydrous CH₂Cl₂To a solution in (5ml) was added TFA (5 ml). After stirring the solution at room temperature for 0.5 h, concentration under reduced pressure gave the title compound as a brown oil (0.255g, 100%).

Example 2

Form A was synthesized from amorphous 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy, (2R, 4R) - (9C 1).

1.8g of amorphous 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) (prepared as described in example 150 of US2003/016272787, Bigge et al) was slurried in 100ml of water at room temperature for three days. The solid was filtered off, washed with 50ml of water and dried under vacuum overnight to give 1.44g of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). PXRD and DSC confirmed that the crystalline form is form a.

Example 3

Synthesis of form A from form B

0.62g of form B (example 4) are stirred in 9.3ml of methanol and 3.2ml of water at 50 ℃. The solution was cooled to room temperature at a rate of 5 deg.C/hour. The solid was filtered to give a white solid. The white solid was shown to be crystalline form a by PXRD and DSC.

Example 4

Crystalline form B was synthesized from amorphous 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).

30.19g of amorphous 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) -phenyl ] -4-methoxy-, (2R, 4R) - (9C1) (prepared as described in Bigge et al, example 150 of US 2003/016272787) was heated to reflux in 275ml MeOH. 175ml of water was heated to 80 ℃ and slowly added to a solution of MeOH/1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). The solution was then cooled to room temperature at a rate of 5 deg.C/hour. The solid was filtered off and washed with 50ml 1: 1 MeOH: water washed and dried in a vacuum oven overnight to give 26.49g of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). The solid was determined to be form B by PXRD and DSC.

Example 5

Synthesis of form C from form B

Form C was synthesized from form B, 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1). About 10mg of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) and 2ml ethyl acetate were placed in a vial. Add stir bar and stir at room temperature for 3 weeks. The solid was filtered off and air dried. The solid was determined to be crystalline form C by PXRD and DSC.

Example 6

Synthesis of form C from a mixture of form B and form C

52g of form B were slurried with 0.5g of form C in 400ml EtOAc overnight. The slurry was filtered to give 33.28g of a solid. The solid was confirmed to be form C via melting point and PXRD.

Example 7

Synthesis of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C1) and subsequent isolation as form C.

2g of (2R, 4R) -4-methoxy-pyrrolidine-2-carboxylic acid [ 2-fluoro-4- (2-oxo-pyridin-1-yl) -phenyl]Amide hydrochloride (prepared as in example 1) was stirred with 0.92ml triethylamine in 17ml EtOAc for 90 min. The reaction mixture was filtered through a pad of celite and then filtered through a pad of 14ml EtOAc rinse. To the reaction filtrate was added 8ml of EtOAc containing 0.86g 4-chlorophenyl isocyanate followed by suspension in 0.25ml CH₃10mg of form C in CN. The reaction was stirred for 2 hours, filtered and the solid washed twice with 6.5ml EtOAc. The solid was dried in an oven to give 2.17g of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridyl) phenyl]-4-methoxy-, (2R, 4R) - (9C 1). The solid was form C as determined by melting point.

Example 8

Synthesis of form B from a mixture of form B and form C

At a temperature above 54 ℃, mixing 1: 1 mixture of forms B and C was slurried overnight in EtOAc at a concentration of greater than 25 mg/ml. The mixture was filtered to give a solid. The solid was determined to be crystalline form B by DSC and PXRD.

Example 9

Synthesis of form B from form C

1.38g of form C was heated to 175 ℃ without solvent and incubated for 10 minutes. The solid was cooled to room temperature. The solid was confirmed to be crystalline form B by melting point and DSC.

Claims (14)

1. Crystalline forms of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).
2. 2. A crystalline form having a powder X-ray diffraction pattern substantially as shown in figure 1A, 1B or 1C.
3. 3. The crystalline form of claim 1, having a powder X-ray diffraction pattern comprising peaks at 19.7 and 23.2 degrees 2 Θ, and one or more other peaks at 16.1 or 21.9 degrees 2 Θ.
4. 4. The crystalline form of claim 1, having a powder X-ray diffraction pattern comprising at least one peak at 16.1, 19.7, 21.9, or 23.2 degrees 2 Θ and having one or more solid state NMR chemical shifts at 173.8 or 111.3 ppm.
5. 5. The crystalline form of claim 1, having a powder X-ray diffraction pattern comprising at least one spectral peak, in degrees 2 Θ, at about 18.9, about 25.9, about 26.0, about 28.7, or about 34.8.
6. 6. The crystalline form of claim 5, having one or more solid state NMR chemical shifts at 172.9 or 110.0 ppm.
7. Crystalline form C of 1, 2-pyrrolidinedicarboxamide, N1- (4-chlorophenyl) -N2- [ 2-fluoro-4- (2-oxo-1 (2H) -pyridinyl) phenyl ] -4-methoxy-, (2R, 4R) - (9C 1).
8. 8. The crystalline form of claim 1 having a powder X-ray diffraction pattern comprising at least one spectral peak, in terms of 2 Θ, at about 13.5 or 17.6.
9. 9. The crystalline form of claim 9, having a powder X-ray diffraction pattern comprising peaks at 13.5 and 17.6 degrees 2 Θ, and one or more other peaks at 9.2, 18.3, or 22.5 degrees 2 Θ.
10. 10. The crystalline form of claim 9, having a powder X-ray diffraction pattern comprising peaks, in terms of 2 Θ, at 13.5 and 17.6 and having one or more solid state NMR chemical shifts at 174.3, 105.4, or 130.3 ppm.
11. 11. The crystalline form of claim 9, having a powder X-ray diffraction pattern comprising at least one peak at 9.2, 13.5, 17.6, 18.3, or 22.5 degrees 2 Θ and having one or more solid state NMR chemical shifts at 174.3, 105.4, or 130.3 ppm.
12. 12. A composition comprising a pharmaceutically acceptable excipient, diluent or carrier, and a therapeutically effective amount of the crystalline form of claim 2, or a mixture thereof.
13. 13. The composition of claim 13, further comprising one or more of the following agents:

    a) non-steroidal anti-inflammatory drugs;

    b) a thrombin inhibitor;

    c) a factor VIIa inhibitor;

    d) a platelet aggregation inhibitor;

    e) a vitamin K antagonist;

    f) a GPIIbIIIa antagonist;

    g) a heparinoid; and

    h) thrombolytic or fibrinolytic agents.
14. 14. A method of treating the following disorders in a mammal with a therapeutically effective amount of the crystalline form of claim 2 or a mixture thereof: venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, cerebral infarction, restenosis, atherosclerosis, angina pectoris, primary deep vein thrombosis, secondary deep vein thrombosis, cancer, sepsis, diabetes or thromboembolism associated with cardiovascular disease.