Crystalline (3z,5s)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one o-methyloxime useful in methods for treating diseases related to OT-r activity

Abstract

To provide efficient and orally selective OT-R antagonist for the treatment of diseases related to the OT-R activity, in particular, preterm labor.SOLUTION: Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is disclosed. Also disclosed are a pharmaceutical composition containing the crystalline compound, and multiple methods for treating multiple conditions related to the OT-R activity, such as preterm labor, and for increasing embryo implantation rate in a mammal undergoing embryo transfer, the methods comprising administering the crystalline compound.SELECTED DRAWING: Figure 4

Description

[Technical field]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 020,076, filed July 2, 2014, the contents of which are incorporated herein by reference in their entirety. This disclosure relates to crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and methods of using same. Additionally, the disclosure relates to pharmaceutical compositions comprising crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, methods for producing same, and methods for treating disorders related to OT-R activity, particularly preterm labor, with crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. [Background technology]

[0002] Oxytocin (OT) is a cyclic nonapeptide that mediates its physiological actions through activation of the oxytocin receptor (OT-R), a cell membrane receptor belonging to the class of G protein-coupled receptors similar to the arginine vasopressin receptor. Among its other roles in the body, OT causes contractions of the mammalian uterus during labor. Repeated, coordinated, and regular contractions of the uterus cause dilation of the cervix and rupture of the fetal membranes, resulting in expulsion of the fetus. Preterm labor is when these contractions occur before the normal expected date of delivery of the pregnancy. A premature increase in uterine activity is the most common sign of preterm labor.

[0003] Premature birth causes unwanted premature birth and remains a serious health problem that is a major cause of perinatal mortality and significant morbidity, especially respiratory distress syndrome, intraventricular hemorrhage, bronchopulmonary dysplasia, and necrotizing enterocolitis, which are much more common in premature infants than in full-term infants. Long-term disabilities such as cerebral palsy, visual impairment, and hearing impairment are also more common in premature infants. Today, premature birth remains a major cause of fetal mortality and morbidity in developed countries. Neonatal intensive care for premature infants is very costly. When considering the provision of health care for premature birth-related illnesses such as respiratory distress syndrome, cardiac disease, cerebral palsy, epilepsy, and severe learning disabilities, the actual costs are much higher for society. Thus, management of premature labor is a major challenge in the field of obstetrics.

[0004] The OT / OT-R system plays a key role in the initiation of labor in mammals, especially in humans. The density of OT-R increases significantly in the myometrium before and during the onset of labor. It is also believed that in humans, local OT peptide hormone concentrations increase significantly before parturition. High blood concentrations of progesterone induce uterine quiescence, while the uterus gains contractility. Just before the expected day of labor, plasma progesterone concentrations fall, uterine OT-R expression increases, OT is released, and uterine contractile activity increases. On the day of parturition, contractions become progressively stronger, culminating in parturition as a result of two interacting positive feedback loops. The first is a local uterine loop in which contractile prostaglandins are produced and released within the uterus itself in response to OT and uterine contractions. These prostaglandins may play further roles in cervical ripening and attenuation of the fetal membranes. The second loop involves the hypothalamus, which responds to uterine contractions and vaginal and cervical distension by causing magnocellular oxytocin neurons in the hypothalamus to increase their activity, leading to the release of OT from their axon terminals in the posterior pituitary gland, which acts on the uterus, stimulating further production of prostaglandins and further contributing to uterine contractions.

[0005] Another possible advantage of antagonizing OT-R is in the field of assisted reproductive technology (ART). Although many efforts have been made over the past decades to improve the outcome of assisted reproduction, the overall effectiveness of in vitro fertilization (IVF) techniques remains limited. Various factors can affect the success rate after IVF. The transfer of the embryo is the key factor influencing the outcome of infertility treatment. ART first involves performing controlled ovarian hyperstimulation (COH) to stimulate the growth of some follicles, allowing the recovery of some follicular cells for IVF. COH is associated with supraphysiological estrogen levels and has been shown to increase uterine contractile activity at the time of embryo transfer in IVF patients compared to naturally occurring menstrual cycles.

[0006] Since uterine contractile activity plays an important role in embryo implantation, uterine contraction is regarded as one of the most fundamental elements of uterine receptivity. Excessive uterine contraction may reduce the embryo implantation rate in IVF cycles because the contractile activity may expel the embryo from the uterus. So far, several therapeutic strategies used to reduce uterine contraction before embryo transfer, such as the use of beta agonists or nonsteroidal anti-inflammatory drugs, have not provided sufficient benefit.

[0007] In addition, systemic and endometrial levels of OT, as well as OT-R expression, are strongly influenced by estrogen, e.g., the highest levels of expression of OT-R are observed mid-cycle in non-pregnant women and in the final month of pregnancy in pregnant women.

[0008] For at least these reasons, it is believed that reducing uterine contractions at the time of embryo transfer by administration of OT-R and / or V1a antagonists may increase embryo implantation rates and, therefore, pregnancy rates in ART.

[0009] Therefore, blocking the effects of OT by antagonizing the OT-R may be an attractive therapy for the treatment of several diseases related to OT-R activity, particularly preterm labor and embryo implantation failure due to uterine contractions.

[0010] Tocolytics, i.e., uterine relaxants, have been used in clinical studies for the medical treatment of preterm labor. Most of these drugs are used off-label. They have shown very limited efficacy, if any, at longer gestational ages and have not shown any clear evidence of improved neonatal outcome. In addition, many tocolytics are often associated with undesirable adverse effects on the woman, fetus, or neonate. Such tocolytics include beta-2-adrenergic agonists, prostaglandin synthesis inhibitors, magnesium sulfate, nitrate donors, and calcium channel blockers. Beta-2-adrenergic agonists, such as ritodrine and terbutaline, produce numerous cardiovascular and metabolic side effects, including maternal tachycardia, palpitations, hypotension, altered thyroid function, and fetal and neonatal hypoglycemia and tachycardia.

[0011] Nifedipine, a calcium channel blocker, is also used in an attempt to stop the contractions. Some possible side effects of this drug include flushing, headache, nausea, palpitations, and lightheadedness. The total prostaglandin synthesis inhibitor (NSAID) indomethacin has also been used, but it too can have serious effects on the fetus, such as ductus arteriosus constriction, pulmonary hypertension, reduced renal function with oligohydramnios, intraventricular hemorrhage, hyperbilirubinemia, necrotizing enterocolitis, and side effects on the mother, such as abdominal discomfort, nausea, vomiting, depression, and dizziness attacks. Sulindac, another NSAID, has a similar side effect profile to indomethacin. A meta-analysis conducted on magnesium sulfate did not support it as a tocolytic agent. Women reported side effects such as flushing, lethargy, headache, muscle weakness, pulmonary edema, and cardiac arrest. Additionally, newborns exposed to magnesium sulfate can show lethargy, hypotension, respiratory depression, bone problems, osteopenia, and fractures. The FDA is now advising health care professionals not to give women magnesium sulfate injections for more than five to seven days to stop premature labor.

[0012] Another drug, atosiban, an antagonist of both the vasopressin V1a receptor and the OT-R, is marketed in Europe and used to stop contractions and delay premature labor by a few days. Atosiban is a peptide that is not orally bioavailable and must be administered parenterally. It is rapidly degraded by enzymes in the circulation, limiting its use to a maximum of 48 hours.

[0013] Selective, orally active small molecule antagonists for the OT-R have been developed in an attempt to overcome these problems. In particular, non-peptide OT-R antagonists have been developed, such as several pyrrolidine derivatives (WO 01 / 72705, WO 02 / 102799, WO 2002 / 074741, WO 2004 / 005249).

[0014] Pyrrolidine derivatives, including a mixture of isomers, have been disclosed for use as oxytocin antagonists (WO 2004 / 005249). Suitable conditions for crystallization of pure (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime or the crude isomeric mixture were not reported therein.

[0015] There remains a significant unmet need for efficient, orally selectable OT-R antagonists for the treatment of diseases related to OT-R activity, particularly preterm labor, In particular, there is a need for orally available, pharmacologic effective products that are rapidly absorbed, have a half-life long enough to accommodate once-daily dosing, and are safe for the mother and fetus in long-term maintenance treatment over the several weeks up to the end of pregnancy. Summary of the Invention

[0016] One embodiment relates to crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, in embodiments of which the crystalline compound may have an XRPD pattern essentially as shown in Figure 1, a DSC curve essentially as shown in Figure 6 or Figure 7, or a TGA curve essentially as shown in Figure 8 or Figure 9.

[0017] Another embodiment is a pharmaceutical composition comprising crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and a pharma- ceutically acceptable excipient. Yet another embodiment is a method for treating a disorder related to OT-R activity comprising administering to a subject a therapeutically effective amount of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. The disorder related to OT-R activity may be selected from the group consisting of premature labor, premature birth, dysmenorrhea, premature ejaculation, sexual dysfunction, endometriosis, embryo implantation failure due to uterine contractions, infertility, benign prostatic hyperplasia, neuropsychiatric disorders, autism, social behavior disorders, psychosocial stress, and cardiovascular disorders. [Brief description of the drawings]

[0018] [Figure 1] Figure 2 shows X-ray powder diffraction (XRPD) analysis for the as-synthesized crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime.

[0019] [Diagram 2]Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (top line) prepared by ethyl acetate crystallization, crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (middle line) prepared by diethyl ether crystallization, vacuum dried on a small scale at ambient temperature for 20 hours and further vacuum dried at 45°C for 65 hours. A comparative XRPD analysis for the O-methyl oxime (bottom trace) is shown.

[0020] [Diagram 3] Figure 2 shows the observed peaks of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime prepared by small scale diethyl ether crystallization and dried under vacuum at ambient temperature for 20 hours with output generated using TriadsTM v2.0.

[0021] [Figure 4] FIG. 1 shows the XRPD pattern of as-synthesized crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime compared to the XRPD pattern of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime after stirring in pH 7 phosphate buffer for 24 hours.

[0022] [Diagram 5]FIG. 1 shows the XRPD pattern of as-synthesized amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime compared to the XRPD pattern of amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime after stirring in pH 7 phosphate buffer for 24 hours.

[0023] [Figure 6] This is a differential scanning calorimetry (DSC) curve of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime prepared by crystallization from diethyl ether.

[0024] [Figure 7] FIG. 1 is a DSC curve of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2′-methyl-1,1′-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime prepared by crystallization from ethyl acetate.

[0025] [Figure 8] Thermogravimetric analysis (TGA) curve of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime prepared by small scale crystallization from diethyl ether.

[0026] [Figure 9] FIG. 2 is a TGA curve of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2′-methyl-1,1′-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime prepared by crystallization from ethyl acetate. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] Crystalline Forms: A novel crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is disclosed herein. Initial attempts to crystallize (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime failed. Over 120 polymorph screening experiments were subsequently performed. More than half of these did not produce solid material suitable for further characterization, and approximately one-third produced an amorphous form. After much experimentation and discovery, the inventors have obtained a stable crystalline form of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, which can be used as a medicament active ingredient in pharmaceutical compositions. The present disclosure teaches how to produce this novel crystalline form and its multiple advantages.

[0028] One embodiment of the present invention is crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime.

[0029] The crystalline compound of the present invention is an essentially pure crystalline compound. An essentially pure crystalline compound is composed mainly of a single crystalline phase, preferably more than about 85% by weight, more preferably more than about 90% by weight, more preferably more than about 95% by weight, even more preferably more than about 98% by weight, and most preferably about 100% by weight of a single crystalline phase. In another embodiment, the crystalline compound is essentially free of amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. Preferably, less than about 8% by weight of amorphous form is present, more preferably less than about 5% by weight of amorphous form is present, and even more preferably less than about 3% by weight of amorphous form is present.

[0030] The term "crystalline" as used herein refers to a compound in the solid state that has a periodic, repeating three-dimensional internal arrangement of the atoms, ions, or molecules characteristic of a crystal. The term crystalline does not necessarily mean that the compound has the appearance of a crystal to the naked eye, but rather that it has this crystal-like internal structural arrangement. The term "amorphous" as used herein refers to a compound that lacks crystalline structure, has only short-range order, without repeating patterns, and is widely disordered.

[0031] Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime can be produced using a solvent of diethyl ether (also referred to as ethyl ether) or ethyl acetate, preferably using diethyl ether. In certain embodiments of the present invention, the amount of residual diethyl ether present after crystallization is less than about 6% by weight, preferably less than about 4% by weight, more preferably less than about 2% by weight. To reduce the amount of residual diethyl ether in the crystalline compound and to avoid melting the solid, the solid can be dried under vacuum at a temperature between about 35°C and about 60°C, preferably between about 40°C and about 55°C, more preferably between about 40°C and about 50°C, and most preferably about 45°C, for a period of at least about 50 hours, preferably at least about 100 hours, more preferably at least about 130 hours. In a particular embodiment, crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is prepared using diethyl ether crystallization and the resulting solid is dried under vacuum at about 45°C for about 133 hours to remove residual ether. Preferably, little manipulation of the solid is required to aid in drying.

[0032] Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime may be used to treat, prevent, or otherwise ameliorate disorders related to OT-R activity.

[0033] Disorders related to OT-R activity include premature labor, premature birth, dysmenorrhea, premature ejaculation, sexual dysfunction, endometriosis, embryo implantation failure due to uterine contractions, infertility, benign prostatic hyperplasia, neuropsychiatric disorders, autism, social behavioral disorders, psychosocial stress, and cardiovascular disorders.

[0034] The term "preterm labor", which also refers to preterm birth, shall mean the expulsion of a viable fetus from the uterus before the normal termination of pregnancy, or more specifically, the onset of labor with effacement and dilation of the cervix before the 37th week of pregnancy, which may or may not be associated with vaginal bleeding or rupture of the membranes.

[0035] The term "dysmenorrhea" refers to a disorder characterized by cyclical pain associated with menstruation during the ovulatory cycle. The pain is thought to result from uterine contractions and ischemia.

[0036] The term "sexual dysfunction" refers to any disturbance or variation in the four phases that characterize the human sexual response: excitement phase, plateau phase, orgasmic phase, and resolution phase.

[0037] As used herein, the term "neuropsychiatric disorder" refers to, for example, depression, obsessive-compulsive disorder and other mental disorders resulting from illnesses of the nervous system.

[0038] As used herein, the term "social behavior disorder" refers to a range of emotional disturbances, inappropriate types of behavior or feelings, pervasive feelings of unhappiness or depression, and perceived difficulties in establishing or maintaining satisfying interpersonal relationships.

[0039] The term "psychosocial stress" as used herein refers to a disorder resulting from a perceived threat to social status, social esteem, self-esteem, respect or acceptance within a group, leading to the development of a stress response in the body and physical symptoms.

[0040] For its use in embryo implantation, infertility, which affects about 10% of human male-female pairs worldwide, can be treated by in vitro fertilization and embryo transfer (IVF-ET) or, in less complicated cases, by artificial insemination. Usually, the success of embryo implantation depends on uterine receptivity, an entity defined as the ability of the uterus to provide optimal conditions for proper implantation and embryo development. The basic elements of uterine receptivity are uterine contractile activity and endometrial disease. Uterine contractions occurring during embryo transfer may expel the embryo from the uterus towards the vagina or fallopian tubes, which may be the cause of an unsuccessful treatment or may later be the cause of ectopic pregnancy, a serious and potentially life-threatening complication. Thus, the crystalline compound may be used in assisted reproduction, more specifically for use by reducing embryo implantation failure due to uterine contractions.

[0041] Common methods for analyzing crystalline forms include crystallography by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA).

[0042] XRPD analyses disclosed herein were collected on an Inel XRG-3000 diffractometer or a PANalytical X'Pert PRO MPD diffractometer.

[0043] An Inel XRG-3000 diffractometer was equipped with a curvilinear position sensitive detector with a 2-theta range of 120°. Real-time data were collected using CuKα radiation starting at approximately 4° 2-theta with a resolution of 0.03° 2-theta. Tube voltage and amperage were set at 40 kV and 30 mA, respectively. Samples were prepared for analysis by packing them into thin-walled glass capillaries. Each capillary was mounted on a goniometer head that was motorized to allow rotation of the capillary during data acquisition. Instrument calibration was performed daily using a silicon reference standard. This setup is shown as the top line in Figure 2, i.e., it was used to collect the XRPD pattern of a crystalline compound produced by ethyl acetate crystallization.

[0044] A PANalytical X'Pert PRO MPD diffractometer was used with an incident beam of CuKα radiation generated using a long fine beam source and a nickel filter, or with an incident beam of Cu radiation generated using an OptiX long fine beam source.

[0045] In the former case, using an incident beam of CuKα alpha radiation generated using a long fine-gas source and a nickel filter, the diffractometer was configured with a symmetric Bragg-Brentano geometry. Prior to the analysis, a silicon specimen (NIST SRM640d) was analyzed to verify that the observed position of the Si111 peak matched the NIST certified position. Specimen samples were prepared as thin circular layers centered on silicon background-free substrates or packed into a backfill holder. Anti-scattering slits (SS) were used to minimize background generated by air. Solar slits for the incident and diffracted beams were used to minimize the spread from axial dispersion. Diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) placed 240 mm from the specimen and data collector software v.2.2b. The data acquisition parameters for each pattern were displayed above the image of the data section, including the divergence slit (DS) and incident beam (SS). This instrument was used to collect the XRPD pattern of the crystalline compound shown in the bottom line of Figure 2, produced by diethyl ether crystallization with small scale vacuum drying at ambient temperature for 20 hours followed by vacuum drying at 45°C for 65 hours.

[0046] In the latter case, using an incident beam of Cu radiation generated with an Optix long fine-gas source, an elliptically graded multilayer mirror was used to focus the CuKα X-rays through the specimen and onto the detector. Prior to analysis, a silicon specimen (NIST SRM640d) was analyzed to verify that the observed position of the Si111 peak matched the NIST certified position. The specimen sample was sandwiched between 3 μm thick membranes and analyzed in transmission geometry. A beam stop, short anti-scattering extension, and anti-scattering knife edges were used to minimize background generated by air. Multiple Soller slits on the incident and diffracted beams were used to minimize spread from axial dispersion. Multiple diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) placed 240 mm from the specimen and data collector software v.2.2b. The data acquisition parameters for each pattern are displayed above the image in the data section, including the divergence slit (DS) in front of the mirror. This setup was used to collect the XRPD patterns shown in Figures 1, 3, 4, 5, and the middle line in Figure 2.

[0047] In certain embodiments, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime has an XRPD pattern after crystallization essentially as shown in Figure 1. In another embodiment, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime has an XRPD pattern essentially as shown in Figure 3.

[0048] As used herein, the term "XRPD pattern" refers to a graphical representation of data collected by XRPD analysis, a technique used to characterize the crystallographic structure, size, and preferred orientation in polycrystalline or powdered solid samples. This diffraction can also be used to characterize heterogeneous solid mixtures to determine the percentage of crystalline compounds present, and provide structural information for unknown materials.

[0049] The terms "essentially" and "about" as used herein in connection with XPRD patterns refer to XPRD patterns in which the recited peaks occur within 0.1 degrees 2-theta, including and including 0.2 degrees 2-theta, of a given 2-theta value.

[0050] "After crystallization" means after preparation of the solid and drying at ambient temperature up to about 45° C. For example, as shown in FIG. 2, the XRPD patterns of a sample dried at ambient temperature for about 20 hours and a sample further dried under vacuum at about 45° C. for about 65 hours are the same. Thus, drying under vacuum at about 45° C. for about 65 hours does not change the solid form of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime.

[0051] In a preferred embodiment, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime has at least characteristic peaks appearing at diffraction angles 2-theta degrees of about 7.05, about 13.13, and about 23.34 as measured by XRPD. In a more preferred embodiment, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime has at least characteristic peaks at diffraction angles 2-theta of about 7.05, about 13.13, about 16.54, about 21.84, and about 23.34 as measured by XRPD, and even more preferably has characteristic peaks at diffraction angles 2-theta of about 7.05, about 12.25, about 13.13, about 16.54, about 18.00, about 21.84, and about 23.34 as measured by XRPD.

[0052] As used herein, the term "characteristic peak" refers to a peak in an XRPD pattern that has an intensity at least 20%, preferably 30% greater than the baseline noise.

[0053] In another embodiment, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime has multiple peaks as measured by XRPD as set forth in Table 1 and shown, for example, in FIG. 3. Table 1: Peaks Observed by XRPD Analysis [Table 1]

[0054] TGA and DSC analyses are used to measure thermal behavior and can be used to distinguish polymorphs. One polymorph may exhibit thermal behavior that is different from that of the amorphous material or from that of another polymorph.

[0055] DSC is a thermoanalytical technique in which the difference between the amount of heat required to increase the temperature of a sample and a reference is measured as a function of temperature. DSC can be used to measure a number of characteristic properties of a sample, allowing the observation of crystallization events. In particular, DSC allows the observation of small energy changes that occur as a material transition from solid to liquid crystal and from liquid crystal to isotropic liquid. The presence of events in the DSC curve can be used to assess the stability of a compound, as well as the presence of solvates or hydrates.

[0056] The DSC analyses disclosed herein were collected on a TA Instrument, Differential Scanning Calorimeter 2920. The instrument was calibrated for energy and temperature using indium. Samples were placed in standard aluminum DSC pans in a no-crimp lid configuration and weights were accurately recorded. The sample cells were equilibrated at 25°C and heated under a nitrogen purge at a rate of 10°C / min up to a final temperature of 350°C. Samples prepared for glass transition determination were heated at 20°C / min in a cyclic experiment. The cyclic experiment consisted of heating the material to either 120°C or 145°C, cooling to 10°C, heating back to that temperature again, cooling to 10°C, and then heating to a final temperature of 350°C.

[0057] TGA is used to determine the change in weight associated with a change in temperature and can reveal degradation of a compound and the presence of solvates or hydrates. The TGA analyses disclosed herein were collected on a TA Instruments 2050 Thermogravimetric Analyzer. Temperature was measured using an Alumel TM and nickel. Each sample was placed in a platinum pan and inserted into a TG furnace. The furnace was heated under a nitrogen purge. Several data acquisition parameters were displayed for each thermogram. The sample whose TGA curve is shown in FIG. 8 was heated from ambient temperature to 350° C. at 10° C. / min. The sample whose TGA curve is shown in FIG. 9 was first equilibrated at 25° C. and then heated to 350° C. at 10° C. / min.

[0058] In certain embodiments, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime exhibits a DSC curve essentially as shown in Figure 6 or Figure 7. In another embodiment, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime exhibits a DSC curve with an endotherm with a signal maximum around about 70°C to about 77°C, a baseline shift at about 122°C to about 130°C, and a large exotherm with a signal maximum at about 230°C to 235°C, possibly due to decomposition. Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime prepared with ethyl acetate may show a DSC curve with a small and broad endotherm with a signal maximum between about 71°C and about 72°C, preferably at about 71.67°C, a baseline shift between about 126°C and about 127°C, preferably at about 127.46°C, and a large exotherm with a signal maximum between about 231°C and about 232°C, preferably at about 231.50°C. Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime prepared with diethyl ether may show a DSC curve with an endotherm with a signal maximum at about 76°C, a baseline shift at about 124°C, and a large exotherm with a signal maximum at about 233°C.

[0059] The term "essentially" as used herein in connection with DSC curves means a DSC curve that exhibits one peak within 1°C, including within 0.5°C, of ​​a given temperature.

[0060] In certain embodiments, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime exhibits a TGA curve essentially as shown in Figure 8 or Figure 9. Preferably, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime analyzed by TGA exhibits a weight loss of about 2% to about 7% at about 25°C to about 200°C, more preferably a weight loss of about 3% to about 6% at about 25°C to about 200°C. In certain embodiments, crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime after crystallization with diethyl ether and analyzed by TGA exhibits a weight loss of about 4.5% to about 5.8%, more preferably a weight loss of about 5.1% to about 5.6%, at about 25°C to about 200°C.

[0061] The term "essentially" as used herein in connection with TGA curves means a curve that shows the percentage of weight loss within 1%, including within 0.5%, of a given value related to temperature change.

[0062] In another embodiment, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime formulated into a tablet or capsule is stable during storage at about 25°C and about 60% relative humidity for at least 6 months, preferably at about 25°C and about 60% relative humidity for at least 12 months, more preferably at about 2°C to about 8°C and ambient humidity for at least 12 months. As used herein, stability during storage means that at least 95% of the crystalline compound is unchanged from the start of the storage period, preferably at least 96% of the crystalline compound is unchanged from the start of the storage period, and most preferably at least 97% of the crystalline compound is unchanged from the start of the storage period.

[0063] The terms "stable" and "stability" as used herein refer to both the physical form and chemical purity of a crystalline compound. As used herein, "crystalline compound" refers to the disclosed crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime of the present invention. Physical stability can be measured by XPRD.

[0064] Ambient conditions as used herein refer to a temperature of about 20° C. to about 25° C. and a relative humidity (RH) of about 40%. Pharmaceutical Compositions

[0065] One embodiment of the present invention relates to a pharmaceutical composition comprising crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and a pharma- ceutically acceptable excipient.

[0066] The pharmaceutical composition of the present invention comprises an effective amount of the crystalline compound and a pharma- ceutical acceptable excipient, and in some embodiments, it may also contain one or more additional active ingredients. The content of the crystalline compound in the pharmaceutical composition of the present invention varies depending on the subject, the route of administration, and the target disease, among other variables. The pharmaceutical composition of the present invention may be administered orally, topically (e.g., transdermally), vaginally, rectally, or parenterally (e.g., intravenously). In one embodiment, the pharmaceutical composition is administered orally.

[0067] Examples of topical administration of pharmaceutical compositions include transdermal, buccal or sublingual administration. For topical administration, the pharmaceutical composition can be suitably mixed into a pharma- ceutically inert topical carrier such as a gel, ointment, lotion, or cream. Such pharma-ceutically inert topical carriers include water, glycerol, alcohol, propylene glycol, fatty alcohol, triglyceride, fatty acid ester, or mineral oil. Other possible pharma-ceutically inert topical carriers are liquid petrolatum, isopropyl palmitate, polyethylene glycol, ethanol 95%, aqueous solution of polyoxyethylene monolaurate 5%, aqueous solution of sodium lauryl sulfate 5%, and the like. In addition, materials such as antioxidants, humectants, viscosity stabilizers, and the like may also be added.

[0068] For oral administration, the crystalline compound may be administered as capsules, tablets, or granules. Tablets may contain various excipients such as microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate, and glycine, along with various disintegrating agents such as starch (preferably corn, potato, or tapioca starch), alginic acid, and certain silicate complexes, as well as granulation binders such as polyvinylpyrrolidone, sucrose, gelatin, and acacia. In certain embodiments, tablets may be coated. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often very useful for tablets. Several other solid compositions may also be used as fillers in gelatin capsules. Preferred materials in this regard also include lactose or milk sugar, and high molecular weight polyethylene glycols. When aqueous suspensions and / or elixirs are desired for oral administration, the crystalline compound may be mixed with various sweeteners or flavorings, coloring substances or dyes, and, if desired, the drug may be emulsified and / or suspended with diluents such as water, ethanol, propylene glycol, glycerin, and various similar combinations thereof. Pharmaceutical compositions may be formulated so that the crystalline compound is released over a period of time after administration.

[0069] Pharmaceutical compositions containing the crystalline compounds in addition to pharma- ceutically acceptable excipients and, optionally, additional active ingredients, may be prepared by any conventional technique known in the art.

[0070] In one embodiment, the amount of the crystalline compound present in the pharmaceutical composition is from about 0.01% to about 90% by weight of the total composition. A suitable therapeutically effective amount of the crystalline compound is typically in the range of about 0.01 mg / kg to about 1 g / kg of body weight per day, in another embodiment, in the range of about 1 mg / kg to about 600 mg / kg of body weight per day, in another embodiment, in the range of about 1 mg / kg to about 250 mg / kg of body weight per day, in another embodiment, in the range of about 10 mg / kg to about 400 mg / kg of body weight per day, in another embodiment, in the range of about 10 mg / kg to about 200 mg / kg of body weight per day, in another embodiment, in the range of about 1 ... In one embodiment, the range is from about 0 mg / kg to about 100 mg / kg of body weight per day, in one embodiment, from about 10 mg / kg to about 25 mg / kg of body weight per day, in another embodiment, from about 1 mg / kg to about 10 mg / kg of body weight per day, in another embodiment, from about 0.001 mg / kg to about 100 mg / kg of body weight per day, in another embodiment, from about 0.001 mg / kg to about 10 mg / kg of body weight per day, in another embodiment, from about 0.001 mg / kg to about 10 mg / kg of body weight per day. In certain embodiments, when the pharmaceutical compositions described herein are orally administered, a suitable therapeutically effective amount of the crystalline compound is from about 0.01 to about 100 milligrams per kilogram of the recipient's body weight per day, preferably from about 0.1 to about 50 milligrams per kilogram of the recipient's body weight per day, and more preferably from about 0.1 to about 20 milligrams per kilogram of the recipient's body weight per day. The desired dose may be administered once a day or in several sub-doses, e.g., two to five sub-doses, at appropriate intervals throughout the day or on any other suitable schedule.

[0071] The term "pharmaceutical acceptable excipients" as used herein includes, but is not limited to, one or more of polymers, resins, plasticizers, fillers, lubricants, diluents, binders, disintegrants, solvents, co-solvents, surfactants, buffer systems, preservatives, sweetening agents, flavoring agents, pharmaceutical grade dyes or pigments, chelating agents, viscosity modifiers, and combinations thereof. Pharmaceutically acceptable excipients may be used in any component for making the dosage form, i.e., the core tablet or the coating. Flavoring agents and dyes and pigments among those useful herein include, but are not limited to, those described in the Handbook of Pharmaceutical Excipients (Pharmaceutical Press, 4th Edition, 2003). Suitable co-solvents include, but are not limited to, ethanol, isopropanol, acetone, and combinations thereof. Suitable surfactants include, but are not limited to, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene monoalkyl ethers, sucrose monoesters, simethicone emulsion, sodium lauryl sulfate, Tween® 80, and lanolin esters, ethers, and combinations thereof. Suitable preservatives include, but are not limited to, phenol, alkyl esters of parahydroxybenzoic acid, benzoic acid and its salts, boric acid and its salts, sorbic acid and its salts, chlorbutanol, benzyl alcohol, thimerosal, phenylmercuric acetate, and nitric acid, nitromersol, benzalkonium chloride, cetylpyridinium chloride, methyl paraben, propyl paraben, and combinations thereof. Suitable fillers include, but are not limited to, starch, lactose, sucrose, maltodextrin, and microcrystalline cellulose. Suitable plasticizers include, but are not limited to, triethyl citrate, polyethylene glycol, propylene glycol, dibutyl phthalate, castor oil, acetylated monoglycerides, triacetin, and combinations thereof.Suitable polymers include, but are not limited to, ethyl cellulose, cellulose acetate trimellitate, hydroxypropylmethylcellulose phthalate, cellulose acetate phthalate, polyvinyl acetate phthalate, and Eudragit® L30-D, Eudragit® L100-55, Eudragit® F530D, and Eudragit® S100 (Rohm Pharma GmbH and Co. KG, Darmstadt, Germany), Acryl-EZE® and Sureteric® (Colorcon, Inc. West Point, Pa.), and combinations thereof. Suitable lubricants include, but are not limited to, magnesium stearate, stearic acid, talc, and combinations thereof.

[0072] The term "additional active ingredient" as used herein includes any agent known in the art for treating, preventing, or reducing the symptoms of the disease being treated by the pharmaceutical composition. Such agents include, but are not limited to, agents known for treating, preventing, or reducing uterine contractions or premature labor, such as calcium channel blockers, magnesium sulfate, selective prostaglandin modulators, beta-2-adrenergic agonists, beta-3-adrenergic receptor agonists, corticosteroids, and mixtures thereof.

[0073] Alternatively, the crystalline compound may be administered in combination with or separately from at least one compound selected from the group consisting of calcium channel blockers (such as nifedipine), magnesium sulfate, prostaglandin receptor modulators (such as agonists or antagonists of any of the EP1, EP2, EP3, EP4, or FP receptors), prostaglandin synthesis inhibitors (such as indomethacin, nimesulide, sulindac, rofecoxib, celecoxib), beta-2-adrenergic agonists (such as ritodrine, terbutaline, salbutamol), beta-3-adrenergic receptor agonists, nitric oxide donors (such as glyceryl nitrate), corticosteroids (such as dexamethasone, betamethasone), and mixtures thereof. As used herein, "combined" refers to administration of the crystalline compound immediately followed or followed by administration of at least one compound. As used herein, "separately" includes sequential or subsequent administration and refers to administration of the crystalline compounds followed by administration of at least one other compound after a period of withdrawal.

[0074] The pharmaceutical composition may be used to treat diseases related to OT-R activity. Certain embodiments relate to pharmaceutical compositions comprising crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and a pharma- ceutical acceptable excipient for use in the treatment of preterm labor. [Method of Use]

[0075] One embodiment of the invention relates to a method for treating a disease associated with OT-R activity comprising administering to a subject a therapeutically effective amount of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. In one embodiment, the invention relates to a method for treating preterm labor.

[0076] The term "treatment" as used herein includes therapeutic and / or prophylactic treatment of a disease related to OT-R activity. Treatment includes the attenuation or alleviation of at least one symptom associated with a disease related to OT-R activity or with another disease described herein.

[0077] The term "therapeutically effective amount" as used herein means an amount of a compound or composition high enough to significantly positively alleviate the symptoms and / or disease to be treated, but low enough to avoid serious side effects (at a reasonable risk / benefit ratio) within the bounds of sound medical judgment. The therapeutically effective amount of a crystalline compound is selected according to a variety of factors, including the type, race, age, weight, sex, and health of the patient, the severity of the disease to be treated, the route of administration, and the renal and hepatic function of the patient. A physician skilled in the art can easily determine and prescribe the effective amount of the drug required to prevent, counter or stop the progression of the disease.

[0078] The term "subject" as used herein is an animal. A "subject" includes, but is not limited to, a human, a mouse, a rat, a guinea pig, a dog, a cat, a horse, a cow, a pig, a monkey, a chimpanzee, a baboon, or a rhesus monkey. In one embodiment, a "subject" is a mammal. In another embodiment, a "subject" is a human, preferably a human female, more preferably a human female of childbearing age.

[0079] In one embodiment, the therapeutically effective amount of the crystalline compound is administered in a single dose, the single dose being between about 10 mg and about 1000 mg, preferably between about 50 mg and about 900 mg, more preferably between about 100 mg and about 600 mg. The crystalline compound may be administered at least once a week, once every two weeks, daily, or multiple times per day. It may be administered in a single dose, or the total dose may be administered in 2, 3, or 4 divided doses per day.

[0080] In one embodiment, crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is administered orally, topically, vaginally, rectally or parenterally, preferably it may be administered intravenously or orally, more preferably it may be administered orally.

[0081] For premature labor treatment, the crystalline compounds may rapidly reduce the frequency and stop uterine contractions for about 2 to about 30 minutes following their administration, preferably for about 5 to about 20 minutes following their administration.

[0082] Another embodiment of the present invention relates to a method for increasing embryo implantation rate in a female mammal during embryo transfer comprising administering to the subject a therapeutically effective amount of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, in a preferred embodiment thereof, the mammal is a human, more preferably a human female of childbearing age.

[0083] "Embryo implantation rate" refers to the number of embryos that attach to the endometrium of a mammal following fertilization produced using assisted reproductive technology (ART). A high embryo implantation rate refers to a high likelihood that an embryo, when transferred to the uterus, will be able to implant in the uterine environment and result in a viable fetus.

[0084] Embryo transfer is understood as related to ART: it is a reproductive technique in which an embryo is produced in the laboratory by fertilization with an egg of a recipient female or a separate donor female and then implantation into the uterus of a recipient female mammal.

[0085] The definitions and preferred embodiments disclosed above for the other embodiments are the same as for this embodiment.

[0086] A further embodiment of the present invention relates to a method for preparing a crystalline compound of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, comprising the steps of: 1) dissolving essentially pure Z-form (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime in a solvent selected from the group consisting of ethyl acetate and diethyl ether; 2) forming crystals; and 3) removing the solvent and concentrating the solution to dryness to provide the formed crystals.

[0087] When essentially pure Z-form (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is contacted with ethyl acetate, the suspension is then 1) sonicated in an ambient bath for about 60 minutes to provide a clear solution, which is further sonicated for about 10 minutes and cooled, 2) crystals are allowed to form at a temperature between -15°C and -25°C for about 4 days, and 3) the solvent, ethyl acetate, is then decanted and the solid is washed with hexane and dried under a nitrogen flow for about 30 minutes.

[0088] When essentially pure Z-form (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is contacted with diethyl ether, the solid is then dissolved by swirling leaving a clear solution at 18-23°C, 2) crystals are allowed to form at about 18°C ​​to 23°C for about 20 hours, and 3) the crystals are isolated by filtration and dried under nitrogen at about 18°C ​​to 23°C for about 4 hours to remove the solvent diethyl ether.

[0089] The following examples illustrate the practice of the invention in some of its preferred embodiments. Other embodiments within the scope of the claims will be apparent to those of ordinary skill in the art.

[0090] [Example] The following examples provide the synthesis of several compounds described herein. Synthesis and Purification of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (amorphous form) [ka]

[0091] The synthesis and purification of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is described in European Patent Application No. 13183723.9, the contents of which are incorporated herein by reference in their entirety. 1.1 Synthesis of (3Z / E,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime

[0092] (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is obtained as a crude isomeric mixture containing (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and (3E,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. The synthesis routes of some of the compounds of the invention are described, for example, in WO 2004 / 005249 and WO 2005 / 082848.

[0093] "(3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime" is also defined by the nomenclature used as "(4Z,2S)-2-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl-carbonyl)]pyrrolidin-4-one O-methyloxime-".

[0094] For example, the compound of the present invention (3Z / E,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime can also be prepared according to steps 1 to 7 described below.

[0095] Stage 1: Preparation of 4-(2-methylphenyl)benzoic acid [ka]

[0096] An aqueous solution (2.20 L, 5.0 vol) of potassium carbonate (0.908 Kg, 6.57 mol, 2.06 wt) was charged to a slurry of an aqueous solution (4.41 L, 15.0 vol) of 4-bromobenzoic acid (0.441 Kg, 2.19 mol, 1.0 wt) at 15° C. to 25° C. The resulting slurry was stirred at 15° C. to 25° C. and degassed three times using vacuum-nitrogen purge cycles. Tetrakis(triphenylphosphine)palladium(0) (0.022 Kg, 0.019 mol, 0.05 wt) was charged and the vacuum-nitrogen purge cycle was repeated. A solution of o-tolylboronic acid (0.313 Kg, 2.30 moles, 0.707 wt) in methanol (3.53 L, 8.0 vol) was degassed three times using vacuum-nitrogen purge cycles and then charged to the slurry of 4-bromobenzoic acid at 15° C. to 25° C. 1The reaction mixture was heated to reflux (71° C. to 78° C.) and maintained at that temperature until completion (reaction deemed complete at 95% conversion), which was typically 1.5 to 2.5 hours, as judged by H NMR analysis (d6-DMSO). The reaction mixture was concentrated to 15 volumes under vacuum at 40° C. to 45° C. Toluene (4.41 L, 10.0 volumes) and tetrahydrofuran (4.41 L, 10.0 volumes) were added to the residue, the resulting mixture was vigorously stirred and acidified to pH 1 with hydrochloric acid (6 M, 2.00 L, 4.5 volumes). The contents were vigorously stirred for 30 to 60 minutes, and the layers were separated. Toluene (2.20 L, 5.0 volumes) and tetrahydrofuran (2.20 L, 5.0 volumes) were added to the aqueous phase, and the mixture was stirred for 5 to 10 minutes. The layers were separated and the combined organic phase was filtered and concentrated under vacuum to 10.0 vol. at 35° C. to 40° C. Toluene (4.41 L, 10.0 vol.) was added to the residue and the reaction product was concentrated under vacuum at 35° C. to 40° C. The tetrahydrofuran content of the resulting slurry was 1 The solid was dried in a vacuum oven at 35°C to 40°C and purified as 4-(2-methylphenyl)benzoic acid [0.438Kg, 94.1%th, 99.3%w / w, 1.0%w / w, 1.2 ... 1 H NMR (d6-DMSO) was consistent with the structure.

[0097] Stage 2: Preparation of 4-(2-methylphenyl)benzoyl chloride [ka]

[0098] Thionyl chloride (0.300 L, 4.11 moles, 0.685 vol) was added to a slurry of 4-(2-methylphenyl)benzoic acid (0.435 Kg, 2.05 moles, 1.0 wt) in toluene (4.35 L, 10.0 vol) at 10° C. to 25° C., and the mixture was 1 The reaction was heated to and maintained at 75-80°C for a period of time usually 4-5 hours until complete by H NMR analysis (d6-benzene). Completion of the reaction was accompanied by the formation of a hazy solution. The reaction product was concentrated to 5.0 volumes by removal of toluene under reduced pressure at 35-45°C. Toluene (2.18 L, 5.0 volumes) was added to the concentrate and the mixture was concentrated to 4.0 volumes by removal of toluene under reduced pressure at 35-45°C. The reaction product was filtered through glass fiber paper and the filter cake was washed with toluene (0.44 L, 1.0 volume). 4-(2-methylphenyl)benzoyl chloride [0.439 Kg, 92.8% th, 100.9% w / w, 1 A toluene solution of [1H NMR (d6-benzene) consistent with structure] was used directly in Stage 3.

[0099] Stage 3: Preparation of (4R)-4-hydroxy-1-[(2'-methyl-1,1'-biphenyl-4-yl)-carbonyl]-L-proline [ka]

[0100] An aqueous solution (0.57 L, 1.3 vol) of potassium carbonate (0.526 Kg, 3.81 mol, 1.2 wt) was charged to a solution of 4-hydroxy-L-proline (0.274 Kg, 2.09 mol, 0.625 wt) in tetrahydrofuran (2.20 L, 5.0 vol) and water (0.44 L, 1.0 vol) at 15° C. to 25° C., followed by a line rinse of water (0.44 L, 1.0 vol). The mixture was cooled to 0° C. to 5° C. with rapid stirring, and a solution of 4-(2-methylphenyl)benzoyl chloride (0.438 Kg, 1.90 mol, 1.0 wt) in toluene (2.19 L, 5.0 vol) was charged at that temperature, followed by a line rinse of toluene (0.44 L, 1.0 vol). The reaction mixture was warmed to 15° C. to 25° C. over 1 to 2 hours and stirred at this temperature until judged complete by TLC analysis. Water (2.20 L, 5.0 vol) was charged to the reaction mixture at 15° C. to 25° C. and the layers were separated. The aqueous phase was acidified to pH 5 to 6 with aqueous hydrochloric acid (6 M, 0.66 L, 1.5 vol) at 15° C. to 25° C., then acidified to pH 1 with aqueous hydrochloric acid (2 M, 0.88 L, 2.0 vol). The mixture was cooled and aged to 0° C. to 5° C. for 30 to 60 minutes, the precipitated solid was collected by filtration, and the filter cake was washed with water (2×1.75 L, 2×4.0 vol) and toluene (0.88 L, 2.0 vol) and sucked dry on the filter for 12 to 24 hours. The collected solid was dried under vacuum at 40° C. to 45° C. until the moisture content by KF was less than 0.2% w / w to give (4R)-4-hydroxy-1-[(2′-methyl-1,1′-biphenyl-4-yl)carbonyl]-L-proline [0.599 Kg, 97.0% th, 136.8% w / w, 1 H NMR (d6-DMSO) was consistent with the structure.

[0101] Stage 4: Preparation of 1-(2'-methyl-1,1'-biphenyl-4-yl)carbonyl-4-oxo-L-proline [ka]

[0102] Triethylamine (1.80 L, 13.56 moles, 3.0 volumes) was charged to a solution of (4R)-4-hydroxy-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]-L-proline (0.598 Kg, 1.84 moles, 1.0 wt) in dimethyl sulfoxide (4.42 L, 7.4 volumes) at 15° C. to 20° C. Pyridine-sulfur trioxide complex (0.879 Kg, 5.52 moles, 1.47 wt) was charged in portions at 15° C. to 25° C. and the reaction mixture was stirred at that temperature until the reaction was complete as judged by TLC analysis (usually 1 to 3 hours). 7 The reaction was quenched with an aqueous solution of hydrochloric acid (3M, 4.80L, 8.0 vol) at 0-30°C, charged with tetrahydrofuran (3.00L, 5.0 vol) and heptane (0.60L, 1.0 vol), the layers were separated and the aqueous phase was extracted with tetrahydrofuran (2x3.00L, 2x5.0 vol). The combined organic phase was washed with an aqueous solution of hydrochloric acid (1M, 2x1.20L, 2x2.0 vol) and saturated sodium chloride solution (2x1.20L, 2x2.0 vol) and the aqueous washes were combined and back extracted with tetrahydrofuran (2x0.60L, 2x1.0 vol). The combined organics were dried over magnesium sulfate (1.794 Kg, 3.0 wt), filtered, the filter cake washed with tetrahydrofuran (0.60 L, 1.0 vol), and the filtrate concentrated under vacuum at 40°-45° C. to give a light brown foam. Ethyl acetate (6.00 L, 10.0 vol) was charged to the foam, the contents stirred for 5-10 minutes until dissolution occurred, and the solvent was removed under vacuum at 40°-45° C. This indicates that tetrahydrofuran is 1The reaction was repeated with ethyl acetate (6.00 L, 5.0 vol) until no H NMR analysis (d6-DMSO) detected. The residue was slurried in ethyl acetate (4.80 L, 8.0 vol) and activated charcoal (0.084 Kg, 0.14 wt) was added followed by a line rinse of ethyl acetate (3.00 L, 5.0 vol) and the reaction mixture was heated to and maintained at 70° C. to 80° C. for 20 to 30 minutes, cooled to 40° C. to 55° C., and filtered through glass fiber paper. The filter cake was washed with ethyl acetate (1.50 L, 2.5 vol) and the combined filtrate and washes were concentrated under vacuum at 40° C. to 45° C. to 2.5 to 3.5 vol.

[0103] Crystallization began during concentration. The concentrate was transferred to a suitable container and heated to 70-80° C. with a line rinse of ethyl acetate (0.30 L, 0.5 vol). Additional ethyl acetate (0.30 L, 0.5 vol) was added as needed to achieve dissolution. Heptane (1.80 L, 3.0 vol) was added at 70-80° C. and the contents were allowed to cool to between 15 and 25° C. over 1-2 hours. The slurry was further cooled and aged to 0-5° C. over 2-3 hours, filtered, and the filter cake was washed with ethyl acetate:heptane (1:1, 0.60 L, 1.0 vol) at 0-5° C., followed by heptane (3.0 L, 2.5 vol). The collected solid was dried under vacuum at 40° C. to 45° C. to give 1-[(2′-methyl-1,1′-biphenyl-4-yl)carbonyl]-4-oxo-L-proline [0.444 Kg, 74.7% th, 74.2% w / w, 1 H NMR (d6-DMSO) was consistent with the structure.

[0104] Stage 5: Preparation of (4Z / E)-4-methoxyimino-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]-L-proline [ka]

[0105] Triethylamine (0.40 L, 2.85 mol, 0.92 vol) was added to a solution of 1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]-4-oxo-L-proline (0.434 Kg, 1.34 mol, 1.0 wt) in dichloromethane (4.40 L, 10.0 vol) at 10°C to 25°C, followed by a line rinse of dichloromethane (0.43 L, 1.0 vol). Methoxylamine hydrochloride (0.130 Kg, 1.56 moles, 0.30 wt) was added portionwise at 10° C. to 25° C., followed by a line rinse of dichloromethane (0.43 L, 1.0 vol), and the reaction mixture was stirred at 10° C. to 25° C. until completion as judged by TLC analysis (usually 3 to 5 hours, TLC eluent: dichloromethane:methanol:acetic acid (90:10:1); UV visualization). The solvent was removed under vacuum at 35° C. to 40° C., and the reaction product was dissolved in ethyl acetate (4.40 L, 10.0 vol) and washed with an aqueous solution of hydrochloric acid (1 M, 2×2.20 L, 2×5.0 vol). The acidic washes were back extracted with ethyl acetate (2.20 L, 5.0 vol), and the combined organic phases were washed with saturated aqueous sodium chloride solution (3.10 L, 7.0 vol), dried under magnesium sulfate (0.300 Kg, 0.69 wt), filtered, and the filter cake was washed with ethyl acetate (2.20 L, 5.0 vol). The filtrate and washes were combined and concentrated under vacuum at 35° C. to 40° C. to give 4-methoxyimino-1-[(2′-methyl-1,1′-biphenyl-4-yl)carbonyl]-L-proline [0.476 Kg, 100.6% th, 109.6% w / w, 1 1 H NMR (CDCl3) was consistent with the structure.

[0106] Stage 6: Preparation of (4Z / E,2S)-methyl-1-[(2'-methyl-1,1'-biphenyl-4-yl)-carbonyl]-4-methoxyimino pyrrolidine-2-carboxylate [ka]

[0107] Potassium carbonate (0.476 Kg, 3.44 moles, 1.0 wt) was added to a solution of 4-methoxyimino-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]-L-proline (0.475 Kg, 1.35 moles, 1.0 wt) in acetone (4.75 L, 10.0 vol) and the mixture was cooled to 0-10°C. Dimethyl sulfate (0.128 L, 1.35 moles, 0.27 vol) was added at 0-15°C and the mixture was stirred at 15-25°C until the reaction was complete as judged by TLC analysis, usually 3-16 hours. The solvent was removed under vacuum at 40-45°C and the reaction product was partitioned between ethyl acetate (3.80 L, 8.0 vol) and water (3.80 L, 8.0 vol). The layers were separated and the organic phase was washed with saturated aqueous sodium chloride solution (2.85 L, 6.0 vol), dried over sodium sulfate (0.953 Kg, 2.0 wt), and filtered. The filter cake was washed with ethyl acetate (0.48 L, 1.0 vol) and the combined filtrate and washings were concentrated under vacuum at 40° C. to 45° C. The excess ethyl acetate was removed by azeotropic distillation with tetrahydrofuran (2×0.95 L, 2×2.0 vol) under vacuum at 40-45° C. to give (4Z / E,2S)-Methyl-1-[(2′-methyl-1,1′-biphenyl-4-yl)-carbonyl]-4-methoxyiminopyrrolidine-2-carboxylate [0.492 Kg, 99.6% th, 103.6% w / w, 1 1 H NMR (CDCl3) was consistent with the structure.

[0108] Stage 7: Preparation of (3Z / E,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime [ka]

[0109] Lithium borohydride (0.049 Kg, 2.26 moles, 0.1 wt) was added portionwise to a stirred solution of (4Z / E,2S)-methyl-1-[(2'-methyl-1,1'-biphenyl-4-yl)-carbonyl]-4-methoxyiminopyrrolidine-2-carboxylate (0.492 Kg, 1.34 moles, 1.0 wt) in tetrahydrofuran (2.31 L, 4.7 volumes) and methanol (2.31 L, 4.7 volumes) under nitrogen at 0° C. to 30° C. The mixture was stirred at 15° C. to 25° C. until the reaction was complete as judged by TLC analysis (eluent: ethyl acetate, color reagent: ninhydrin), usually for 2 to 6 hours. The reaction mixture was quenched with water (0.40 L, 0.8 vol) at 15-25°C and stirred at 15-25°C for 16-20 hours. The reaction product was concentrated under vacuum at 40-45°C and the residue partitioned between water (2.46 L, 5.0 vol) and ethyl acetate (4.92 L, 10.0 vol). The layers were separated and the organic phase was washed successively with aqueous hydrochloric acid (1M, 2.46 L, 5.0 vol), saturated aqueous sodium bicarbonate solution (2.46 L, 5.0 vol), and saturated aqueous sodium chloride solution (2.46 L, 5.0 vol). The organic phase was dried over magnesium sulfate (0.985 Kg, 2.0 wt), filtered, and the filter cake was washed with ethyl acetate (0.50 L, 1.0 vol). The combined filtrate and washings were concentrated under vacuum to give (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime as a viscous brown oil and (3E,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime [0.395Kg, 86.9%th, 80.3%w / w, 1A crude mixture of isomers was obtained containing 1H NMR (CDCl3) consistent with structure, 82.0% area by HPLC, Z / E ratio of 71.4:28.6. The oil was dissolved in toluene (0.40 L, 1.0 volume relative to the weight of the product) and stored until needed. 1.2 Essentially pure form of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime 1.2.1 Small scale purification

[0110] The isolation step of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime in essentially pure form was carried out by chromatography using a Biotage system (Biotage AB, SE-751 03 Uppsala, Sweden) of the crude isomer mixture isolated after reduction of the oxime ester (stage 7 of Example 1).

[0111] Five separate batches of the crude isomeric mixture (No. 020, 180, 062, 068, 076) were purified by Biotage chromatography. In addition, different conditions were used for batches No. 068 and 076. Purification was performed on a Biotage column (No. 076) spiked with 5% w / w of oxime methyl ester.

[0112] Each chromatography was performed using a Biotage 40M cartridge (40 g silica) that was pre-flushed with toluene:MeOH (99:1 v / v) and then eluted and collected in 100 ml fractions (total volume 4 L) followed by a flush with toluene:MeOH (96:4 v / v).

[0113] Fractions were analyzed by TLC (eluent: ethyl acetate) to determine which fractions could be discarded and which contained the Z isomer. These Z fractions were then analyzed by HPLC. The acceptance criteria for fractions were >96% Z isomer and <1.2% E isomer.

[0114] Surprisingly, purification of the various batches via Biotage chromatography is highly efficient, with the essentially pure form of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime being purified to 99.4% (Batch No. 020, No. 062, No. 068) and 99.2% (Batch No. 180, No. 076). In particular, Biotage chromatography in the presence of oxime ester removes 5% w / w of oxime ester without loss of recovery or quality (Batch No. 068), and 25% overloading of the Biotage column does not cause a reduction in yield or quality (Batch No. 076). Table 2: Efficiency of Biotage chromatography [Table 2]

[0115] 1.2.2 Large Scale Purification Various batches of crude (3Z / E,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (0.392 kg, 1.16 moles, 1.0 wt) were charged to a Biotage 150 L SIM unit as approximately 50% w / w solution in toluene and purified using 1% methanol in toluene (150 L) followed by 2% methanol in toluene (50 L) in fraction sizes of 5.0 L. Collected fractions were analyzed by TLC and HPLC analysis as appropriate. The fractions believed to contain clean (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (criteria: Z isomer ≥ 96.00% area, E isomer ≤ 1.20% area) were combined and concentrated under vacuum at 40-45°C. Anhydrous ethanol (2x2L) was added to the residue and the solution was concentrated under vacuum at 40-45°C until a foamy solid could be worked up. The desired product (3Z,5S)-1-[(biphenyl-4-yl-carbonyl)-5-hydroxy-methyl]pyrrolidin-3-one-O-methyloxime (0.089Kg, 22.7% w / w, 1 H NMR (CDCl3) consistent with structure, 99.3% area by HPLC, Z / E ratio of 98.4:0.9) was obtained as a slightly off-white to light brown solid. Table 3: Summary of different batch purifications of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime in essentially pure form. [Table 3]

[0116] Appropriate batches of (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (2.713 kg, 1.0 wt.) isolated by biotage chromatography were combined, dissolved in absolute ethanol (5.16 L, 2.0 vol.) at 15-25°C, and clarified by filtration through glass fiber paper, with an absolute ethanol wash (0.50 L, 0.2 vol.) applied to the filter. The combined filtrate was concentrated in portions under vacuum at 40-45°C. The reaction product was transferred to a drying tray and dried under vacuum at 30°C for 24 hours. The oven temperature was then gradually increased from 30°C to 40°C for 80 hours. Residual solvent levels were determined to be 0.01% by weight. 1 When the solvent level was found to be less than 1.0% w / w as determined by H NMR analysis (CDCl3), the solid was passed through a 500 μm opening sieve. The solid was returned to the oven to obtain (3Z,5S)-1-[(biphenyl-4-yl-carbonyl)-5-hydroxy-methyl]-pyrrolidin-3-one-O-methyloxime (2.633 Kg, 97.1% w / w ... 1 The solid was dried at 40-42°C until 1 H NMR (CDCl3) consistent with structure, 98.65% area by HPLC was obtained.

[0117] The combined steps are summarized below: Input: 2.713 kg Output: 2.633 kg Yield: 97.1% w / w Synthesis of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime ("Crystalline Compound")

[0118] A series of experiments were performed before successful crystallization was achieved. Unsuccessful crystallization attempts included the use of acetone, ACN, CH2CL2, 1,4-dioxane, ethanol, hexane, IPA, methanol, MEK, TFE, THF, toluene, and water, among other solvents. After each of these experiments, the resulting solid was examined and XRPD analysis showed the compound to be amorphous. 2.1 Crystallization with Ethyl Acetate

[0119] Ethyl acetate was added to the amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. The suspension was sonicated in an ambient bath for 60 minutes to provide a clear solution, which was sonicated for an additional 10 minutes and cooled. The clear solution was placed in a freezer for 4 days. The solvent was decanted and the solid was washed with hexane and then dried under a nitrogen flow for 30 minutes. The resulting solid was analyzed by XRPD and confirmed to be crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. Figures 7 and 9 are the DSC and TGA curves, respectively, obtained by examining this sample. Characterization data are presented in Table 4. Table 4: Characterization data of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime from ethyl acetate [Table 4]

[0120] 2.2 Crystallization with Diethyl Ether Diethyl ether (40 mL) was added to amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (4.0 g). The solid was dissolved by swirling and the clear solution was left at 18-23°C. Spontaneous nucleation occurred after about 10-15 min and the crystallization vessel and contents were allowed to stand undisturbed at 18-23°C for 20 h. The crystals were isolated by filtration and dried under nitrogen at 18-23°C for 4 h. The yield was 3.38 g (84%) of white crystals with a chemical purity of 99.43%.

[0121] The resulting solid was analyzed by XRPD and confirmed to be crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. Figures 6 and 8 are the DSC and TGA curves, respectively, obtained by examining this sample. Characterization data are presented in Table 5. Table 5: Characterization data of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime from diethyl ether [Table 5]

[0122] 2.3 Other crystallization attempts using diethyl ether Numerous attempts to prepare crystalline compounds using diethyl ether were carried out, as listed in Table 6. Table 6: Attempts to prepare crystals from amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime using diethyl ether [Table 6]

[0123] One such preparation involved adding 0.897 ml of diethyl ether to 240 mg of amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and the mixture was left at ambient temperature for approximately 24 hours. The liquid was decanted and the solid was transferred to a drying vessel to air dry for 5 hours. The solid was then vacuum dried at ambient temperature for 20 hours. The resulting solid was analyzed by XRPD and confirmed to be a crystalline compound. The XRPD pattern from this sample is shown in Figure 1.

[0124] 23 mg of the solid was then dried under vacuum at 45° C. for an additional 65 hours. The resulting solid was analyzed by XRPD and confirmed to be a crystalline compound. Characterization data is presented in Table 7. Table 7: Characterization data for crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime from diethyl ether [Table 7]

[0125] inspection Solid State Pressure Experiments Solid state pressure experiments were performed on crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, the results of which are shown in Tables 8 and 9 below. Confirmed by XRPD, the results showed that the crystalline form remained unchanged after pressing the material at 75% RH for 3 days. Pressing at 97% RH for the same time dissolved the material. The sample was also pressed at 10,000 psi for 1 minute, and XRPD showed that it remained in crystalline form. However, hand grinding for 1 minute produced a mixture of crystalline and amorphous compounds, as shown by XRPD. Table 8: Solid-state pressure studies on crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime [Table 8] Table 9: Solid-state pressure study of amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime [Table 9]

[0126] Solution Pressure Experiments Solution pressure experiments were also performed and the results are presented in Table 10. Table 10: Crystallization Solution Pressure Experiments [Table 10]

[0127] Sonication in the ethyl acetate example resulted in material that appeared as grouped blades by light microscopy. After washing with hexane and drying under nitrogen, the material appeared as irregular clumps. The sample was analyzed by XRPD and found to be crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. In another example using diethyl ether, the mixture was frozen before mixing and drying under nitrogen to produce the crystalline form.

[0128] All other slurries did not produce any solid material. Comparative Stability Study

[0129] Slurries of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime and amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime were prepared by weighing directly into vials of 100 mg of crystalline or amorphous material. Approximately 3 ml of pH 7 phosphate buffer USP was added to each sample. Each sample of the slurry was stirred in pH 7 phosphate buffer (USP) at 25°C for 24 hours. At the end of the 24 hours, the solvent was removed from the samples by pipetting out and the solids were allowed to air dry overnight in a ventilated compartment. The solids were then submitted for XRPD analysis. Observations of the samples were taken at various time intervals and are provided in Table 11. Both samples initially appeared as white powders. Images of the samples were taken at time points corresponding to 5 minutes, 15 minutes, 1 hour, 3 hours, and 24 hours after addition of pH 7 phosphate buffer. Table 11: Observations of Samples Stirred in pH 7 Phosphate Buffer at 25° C. [Table 11]

[0130] Amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime turned into an opaque sticky material after about 15 minutes. After about 1 hour, the crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime slurry showed the form of a white sticky material. Analysis of XPRD pattern showed the presence of an amorphous phase after stirring amorphous (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime for 24 hours. For comparison, Figure 5 shows the XRPD pattern of the amorphous form as synthesized and after stirring in pH 7 buffer for 24 hours. Figure 4 shows the XRPD pattern of the crystalline compound as synthesized and after stirring in pH 7 buffer for 24 hours. The XPRD pattern of the solid obtained upon stirring crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime in buffer shows multiple peaks that are consistent with those seen in the powder pattern of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime (Figure 4). When comparing the before and after XRPD patterns of a sample of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, slight peak shifts are seen at angles higher than 2 theta. Some diffuse scattering is also seen in the powder pattern of the solid obtained from a slurry of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime.

[0131] Numerous changes, modifications, and variations of the preferred embodiments disclosed herein will be apparent to those skilled in the art, all of which are anticipated and considered within the spirit and scope of the claimed invention. For example, while several specific embodiments have been described in detail, those skilled in the art will understand that several of the above embodiments and variations can be modified to incorporate various types of substitutions, additions, or alternative materials. Thus, even if only a few variations of the invention are described herein, it will be understood that such additional modifications and variations, and their equivalents, are included within the spirit and scope of the invention as defined in the following claims. All patent applications, patents, and other publications cited herein are incorporated by reference in their entirety. Here, examples of the invention according to this embodiment will be described as items. [Item 1] Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. [Item 2] 2. The crystalline compound according to claim 1, having an XRPD pattern essentially as shown in FIG. [Item 3] Item 1. The crystalline compound according to item 1, having characteristic peaks at diffraction angles (2 theta degrees) of at least about 7.05, about 12.25, about 13.13, about 16.54, about 18.00, about 21.84, and about 23.34 as measured by XRPD. [Item 4] Item 4. The crystalline compound according to item 3, having at least characteristic peaks appearing at diffraction angles (2 theta degrees) of about 23.34, about 13.13 and about 7.05 as measured by XRPD. [Item 5] 5. The crystalline compound according to any one of items 1 to 4, having a DSC curve essentially as shown in FIG. 6 or FIG. 7. [Item 6] 5. The crystalline compound according to any one of items 1 to 4, having a DSC curve with an endotherm with a signal maximum around about 70°C to about 77°C, a baseline shift at about 122°C to about 130°C, and a large exotherm with a signal maximum at about 230°C to 235°C. [Item 7] Item 7. The crystalline compound according to item 6, having a DSC curve with a small and broad endotherm with a signal maximum between about 71°C and about 72°C, preferably at about 71.67°C, a baseline shift between about 126°C and about 127°C, preferably at about 127.46°C, and a large exotherm with a signal maximum between about 231°C and about 232°C, preferably at about 231.50°C. [Item 8] Item 7. The crystalline compound according to item 6, having a DSC curve with an endotherm containing a signal maximum at about 76° C., a baseline shift at about 124° C., and a large exotherm containing a signal maximum at about 233° C. [Item 9] 9. The crystalline compound according to any one of items 1 to 8, having a TGA curve essentially as shown in FIG. 8 or FIG. [Item 10] 9. The crystalline compound according to any one of items 1 to 8, exhibiting a weight loss of about 2% to about 7% at about 25° C. to about 200° C. as analyzed by TGA, more preferably a weight loss of about 3% to about 6% at about 25° C. to about 200° C. [Item 11] 11. The crystalline compound according to item 10, exhibiting a weight loss of about 4.5% to about 5.8%, more preferably about 5.1% to about 5.6%, at about 25° C. to about 200° C. as analyzed by TGA. [Item 12] 12. A pharmaceutical composition comprising a crystalline compound according to any one of items 1 to 11 and a pharma- ceutical acceptable excipient. [Item 13] 13. The pharmaceutical composition according to item 12, for use in the treatment of diseases related to OT-R activity. [Item 14] 14. The pharmaceutical composition according to item 13, wherein the disease is selected from the group comprising premature labour, premature birth, dysmenorrhea, premature ejaculation, sexual dysfunction, endometriosis, embryo implantation failure due to uterine contractions, infertility, benign prostatic hyperplasia, neuropsychiatric disorders, autism, social behavioural disorders, psychosocial stress and cardiovascular disorders. [Item 15] 15. The pharmaceutical composition according to item 14, wherein the disease is preterm labor. [Item 16] 12. A method for treating a disease associated with OT-R activity, comprising administering to a subject a therapeutically effective amount of the crystalline compound according to any one of items 1 to 11. [Item 17] 17. The method according to item 16, wherein the disease is selected from the group comprising premature labour, premature birth, dysmenorrhea, premature ejaculation, sexual dysfunction, endometriosis, embryo implantation failure due to uterine contractions, infertility, benign prostatic hyperplasia, neuropsychiatric disorders, autism, social behavioural disorders, psychosocial stress and cardiovascular disorders. [Item 18] 18. The method of claim 16 or 17, wherein the subject is a human. [Item 19] 12. A method for treating premature labor comprising administering to a subject a therapeutically effective amount of the crystalline compound of any one of items 1 to 11. [Item 20] 20. The method of claim 19, wherein the crystalline compound is administered in a single dose of about 10 mg to about 1000 mg. [Item 21] 21. The method of claim 20, wherein the single dose is from about 50 mg to about 900 mg. [Item 22] 22. The method of any one of items 19 to 21, wherein the subject is a human. [Item 23] 23. The method of claim 22, wherein the subject is a human female. [Item 24] A method for increasing an embryo implantation rate in a subject during embryo transfer comprising administering to the subject a therapeutically effective amount of crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime. [Item 25] 25. The method of claim 24, wherein the subject is a human. [Item 26] 26. The method of claim 25, wherein the subject is a human female. [Item 27] 1. A process for preparing a crystalline compound of essentially pure Z-form (3Z,5S)-5-(hydroxymethyl)-1-[(2′-methyl-1,1′-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime, comprising the steps of: 1) dissolving (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime using a solvent selected from the group consisting of ethyl acetate and diethyl ether; 2) forming a plurality of said crystals; 3) removing the solvent and concentrating the solution to dryness to provide a plurality of the formed crystals; Equipped method. [Item 28] 28. The method of claim 27, wherein 1) the essentially pure Z-form (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is contacted with ethyl acetate and the suspension is sonicated in an ambient bath for about 60 minutes to provide a clear solution, which is further sonicated and cooled for about 10 minutes; 2) the crystals are allowed to form at a temperature between about -15°C and -25°C for about 4 days; and 3) the solvent, ethyl acetate, is then decanted and the solid is washed with hexane and dried with a nitrogen flow for about 30 minutes. [Item 29] 28. The method of claim 27, wherein 1) the essentially pure Z-form (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime is contacted with diethyl ether, the solid is then dissolved by swirling and the clear solution is left at 18°C ​​to 23°C, 2) the crystals are allowed to form at about 18°C ​​to 23°C for about 20 hours, and 3) the crystals are isolated by filtration to remove the solvent diethyl ether at about 18°C ​​to 23°C for about 4 hours and dried under nitrogen.

Claims

[Claim 1] Crystalline (3Z,5S)-5-(hydroxymethyl)-1-[(2'-methyl-1,1'-biphenyl-4-yl)carbonyl]pyrrolidin-3-one O-methyloxime.

Images