CN113896670B

CN113896670B - Salts of pyrrolidine amide derivatives and uses thereof

Info

Publication number: CN113896670B
Application number: CN202110759380.7A
Authority: CN
Inventors: 金传飞; 陈康智
Original assignee: Sunshine Lake Pharma Co Ltd
Current assignee: Guangdong HEC Pharmaceutical
Priority date: 2020-07-07
Filing date: 2021-07-06
Publication date: 2023-06-09
Anticipated expiration: 2041-07-06
Also published as: CN113896670A

Abstract

The present invention relates to salts of pyrrolidine amide derivatives and uses thereof. The invention also relates to a pharmaceutical composition comprising said salt, and the use of said salt or of said pharmaceutical composition of said salt for the preparation of a medicament for the prevention, treatment or alleviation of diseases mediated by MAO-B, including neurodegenerative diseases, in particular Parkinson's disease.

Description

Salts of pyrrolidine amide derivatives and uses thereof

Technical Field

The invention belongs to the technical field of medicines, relates to salts of pyrrolidine amide derivatives and application thereof, in particular to salts of (2S, 4S) -4-fluoro-1- (4- (3-fluorobenzyloxy) benzyl) pyrrolidine-2-carboxamide, crystal forms of the salts and application thereof, and further relates to a pharmaceutical composition containing the salts or the crystal forms of the salts.

Background

Parkinson's Disease (PD) is a common chronic degenerative disease of the nervous system, common to the elderly, and rarely seen in young parkinsonism with an average age of about 60 years and less than 40 years. The prevalence of PD in people over 65 years old in China is about 1.7%. Most parkinson's disease patients are sporadic cases, with less than 10% of patients having a family history. Parkinson's disease is a disease that is insidious and slow in progression. The first symptoms are typically tremors or clumsiness of one limb, which in turn involves the opposite limb. Clinically, it is mainly manifested by resting tremor, bradykinesia, myotonia and postural gait disorder. In recent years, people increasingly pay attention to non-motor symptoms such as depression, constipation, sleep disturbance and the like, which are common complaints of parkinsonism patients, and influence on the life quality of patients even exceeds motor symptoms.

The most important pathological changes of parkinson's disease are degeneration and death of neurons of the midbrain substantia nigra Dopamine (DA), thereby causing significant reduction of striatal DA content to cause disease. The exact etiology leading to this pathological change is currently unknown, and genetic factors, environmental factors, age-related aging, oxidative stress, etc. may be involved in the degenerative death process of PD dopaminergic neurons.

Most cases are likely to be associated with environmental factors or as a result of interactions of environmental factors with genetic factors. Part of the pathogenesis involves free radicals, oxidative stress, glutamate excitotoxicity, lack of neurotrophic agents, inflammation, apoptosis and loss of mitochondrial complex I, and these mechanisms interact in cascade biochemical reactions ultimately leading to neuronal death (Teismann P, schulz jb. Cellular pathology of Parkinson's disease: astrocytotes, microglia and inflammation [ J ]. Cell Tissue Res,2004, 318:149-161). Genetic factors play a decisive role in part of familial PD. Recent genetic studies have found that functional defects of the ubiquitin-proteinase system and abnormal aggregation of denatured proteins play an important role in the pathogenesis of most PD. In addition, factors such as oxidative stress and the formation of free radicals, excitotoxicity mediated by excessive glutamate release, mitochondrial dysfunction, inflammation, and neuronal apoptosis caused by damage to the ubiquitin-protease system are closely related to the progression of the disease.

Currently, the main treatment for PD is symptomatic treatment of dopamine replacement, and L-dopa (L-dopa) is still the most effective drug for controlling PD symptoms and signs clinically (RASCO O, GOETZ C, KORLER W, et al, treatment interventions for Parkinson's disease: an evidence based assessment [ J ]. Lancet,2002,359 (9317): 1589-1598). Although L-dopa is capable of temporarily controlling the symptoms of PD, chronic administration can lead to a number of adverse effects such as catabolism, motor fluctuations and psychotic symptoms. Although the use of sustained DA can neuronal stimulation, deep brain stimulation of surgical pathways (deep brain stimuli, DBS), long acting dopamine receptor agonists can reduce the incidence of these complications to some extent (SCHAPIRA A H V, EMREB M, JENNER P, et al Levodopa in the treatment of Parkinson's disease [ J ]. Eur J Neurol,2009,16 (9): 982-989), does not delay the progression of the disease. In addition, dopamine receptor agonists such as cabergoline (cabergoline), catechol-oxy-methyltransferase inhibitors (COMT) such as entacapone (Comtan), glutamate receptor antagonists such as memantine (memantine), anticholinergic agents such as benzohexol (ambam) all have adverse effects, but can be used as auxiliary drugs of levodopa, and the drug effect of the levodopa can be enhanced, the dosage of the levodopa can be reduced and the adverse effects can be reduced by combining different action mechanisms of the drug. Therefore, research and development of new drugs which can improve the symptoms of DA and non-DA systems of PD patients and slow down or even prevent the progression of diseases to exert neuroprotection are particularly important.

Monoamine oxidase (MAO, EC 1.4.3.4) is a flavin-containing enzyme responsible for oxidative deamination of endogenous monoamine neurotransmitters, including: dopamine, 5-hydroxytryptamine, epinephrine or norepinephrine, and trace amines such as phenylethylamine, many amine xenobiotics, and the like. Monoamine oxidase can be divided into two subtypes, monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B). Their gene codes are different (A.W.Bach et al, proc.Natl.Acad.Sci.USA 1988,85,4934-4938) and also differ in structure, tissue distribution and substrate specificity. MAO-A is present in liver and gastrointestinal mucosA, and can inactivate catecholamine in blood circulation system and vasoactive substances (such as tyrosine) in diet, thereby assisting in degradation of neurotransmitter in brain; MAO-B is found mainly in the brain and in platelets. MAO-A has higher affinity for octopamine, 5-hydroxytryptamine, epinephrine, and norepinephrine; whereas the natural substrates for MAO-B are tyramine and phenylethylamine. Both isoforms, however, oxidize dopamine.

Monoamine oxidase B (monoamine oxidase B, MAO-B) is one of the key enzymes of DA catabolism, and can improve clinical symptoms by selectively and specifically inhibiting endogenous and exogenous dopamine decomposition and prolonging the acting time of dopamine, so that the monoamine oxidase B can be used for early single drug treatment of PD and adjuvant treatment after symptom fluctuation. The main effects are as follows: (1) Decomposition of dopamine to 3, 4-dihydroxyphenylacetic acid And homovanillic acid, and simultaneously producing small molecule H ₂ O ₂ Producing toxic effects on nerve cells; (2) Deamination of beta-phenylethylamine which stimulates dopamine secretion and inhibits dopamine reuptake is deactivated; (3) Also can decompose 1-methyl-4-phenyl-1, 2,3, 6-tetrahydropyrimidine (MPTP) into l-methyl-4-phenylpyridine ion (MPP) with neurotoxicity ⁺ ). Thus, according to the physiological function of MAO-B, on one hand, inhibiting the activity of MAO-B can reduce the degradation and reuptake of dopamine, increase the concentration of dopamine in brain, and improve the clinical symptoms of PD; on the other hand, by reducing H ₂ O ₂ 、MPP ⁺ Isotoxin levels delay the death process of melanocytes (HEIKKILA R E, MANZINO L, CABBAT F S, et al protection against the dopaminergic neurotoxiciy of 1-methyl-1,2,3,6-tetrahydropyridine (MPTP) by monoamine inhibitors [ P)].Nature,1984,311(5985):467-469；YOUDIM M B H,BAKHLE Y S.Monoamine oxidase isoforms and inhibitors in Parkinson’s disease and depressive illness[J].Br J Pharmacol,2006,147(S1):S287-S296；NAOI M,WAKAKO M.Monoamine oxidase inhibitors as neuroprotective agents in age-dependent neurodegenerative disorders[J]Curr Pharm Des,2010,16 (25): 2799-2817) can alter PD progress. MAO-B inhibitors are hot spots in current anti-Parkinson's disease drug research because they not only improve PD symptoms, but also play a role in neuroprotection.

International application WO 201917515 A1 discloses the compound (2 s,4 s) -4-fluoro-1- (4- (3-fluorobenzyloxy) benzyl) pyrrolidine-2-carboxamide (compound of formula (I)) which has a better inhibitory effect on the activity of MAO-B. However, no studies have been made in the prior art on salts of the compound or crystalline forms thereof.

Different salts and solid forms of the pharmaceutically active ingredient may have different properties. Different salts and solid forms may vary significantly in appearance, solubility, melting point, dissolution, bioavailability, etc., and may also have different effects on stability, bioavailability, efficacy, etc. of the drug. Thus, problems with the salt form and/or solid form of the drug should be fully considered in drug development.

The inventor finds that the compound has poor water solubility and poor patent drug property when researching the compound, so in order to find a solid form with better patent drug property, a large amount of experimental researches show that after the compound shown as the formula (I) forms a salt, the physicochemical properties of different salts are greatly changed, and the properties of some salts are not better than those of the compound in a free state; the inventor discovers that the physical properties and various properties of the compound phosphate shown in the formula (I) prepared by the method can be obviously improved, and the preparation development is facilitated.

Disclosure of Invention

The invention provides a salt of a compound shown in a formula (I), and researches on the preparation of the salt, the solid form of the salt, the physicochemical properties and the pharmacological properties of the salt show that the salt formed by the compound and different acids has larger difference in physicochemical properties; wherein various physicochemical properties of the phosphate are better than those of other salts, for example, the phosphate crystal form A obtained after the compound shown in the formula (I) is salified with phosphoric acid has better pharmacokinetic properties than the corresponding hydrochloride crystal form A, maleate crystal form A and p-toluenesulfonate crystal form A. Therefore, the phosphate crystal form A of the invention has better properties and better pharmacokinetic properties, thereby having better patentability.

In particular, the invention relates to salts of the compounds of formula (I) and the use of the crystalline forms of the salts or pharmaceutical compositions comprising the salts or the crystalline forms of the salts in the manufacture of a medicament for the prevention, treatment or alleviation of diseases mediated by MAO-B, including neurodegenerative diseases, in particular Parkinson's disease. The salt is phosphate. Further, the salt is phosphate crystal form A. The crystalline forms of the invention may also be in the form of solvates, for example hydrates.

In one aspect, the present invention provides a salt of a compound of formula (I),

in some embodiments, the salts of the present invention are organic or inorganic acid salts.

In other embodiments, the inorganic acid salts described herein include, but are not limited to, hydrochloride, hydrobromide, phosphate, nitrate, sulfate, or the like; the organic acid salts include, but are not limited to, acetates, oxalates, fumarates, maleates, tartrates, citrates, succinates, camphorsulfonates, malonates, benzoates, salicylates, benzenesulfonates, methanesulfonates, p-toluenesulfonates, and the like.

In some embodiments, the salt of the compound of formula (I) of the present invention is a phosphate salt.

In some embodiments, the phosphate salt of the present invention is phosphate form a, which has an X-ray powder diffraction pattern with diffraction peaks at the following 2θ angles: 4.28 ° ± 0.2 °,12.13 ° ± 0.2 °,12.22 ° ± 0.2 °,12.83 ° ± 0.2 °,13.07 ° ± 0.2 °,16.29 ° ± 0.2 °,17.06 ° ± 0.2 °,18.14 ° ± 0.2 °,18.80 ° ± 0.2 °,20.34 ° ± 0.2 ° and 25.42 ° ± 0.2 °.

In some embodiments, the phosphate salt of the present invention is phosphate form a, which has an X-ray powder diffraction pattern with diffraction peaks at the following 2θ angles: 4.28 ° ± 0.2 °,12.13 ° ± 0.2 °,12.22 ° ± 0.2 °,12.83 ° ± 0.2 °,13.07 ° ± 0.2 °,16.29 ° ± 0.2 °,17.06 ° ± 0.2 °,17.77 ° ± 0.2 °,18.14 ° ± 0.2 °,18.80 ° ± 0.2 °,19.39 ° ± 0.2 °,19.81 ° ± 0.2 °,20.05 ° ± 0.2 °,20.34 ° ± 0.2 °,23.15 ° ± 0.2 °,25.42 ° ± 0.2 ° 26.72 ° ± 0.2 °.

In some embodiments, the phosphate salt of the present invention is phosphate form a, which has an X-ray powder diffraction pattern with diffraction peaks at the following 2θ angles: 4.28 ° ± 0.2 °,12.13 ° ± 0.2 °,12.22 ° ± 0.2 °,12.83 ° ± 0.2 °,13.07 ° ± 0.2 °,16.29 ° ± 0.2 °,17.06 ° ± 0.2 °,17.77 ° ± 0.2 °,18.14 ° ± 0.2 °,18.80 ° ± 0.2 °,19.39 ° ± 0.2 °,19.81 ° ± 0.2 °,20.05 ° ± 0.2 °,20.34 ° ± 0.2 °,22.70 ° ± 0.2 °,23.15 ° ± 0.2 °,23.91 ° ± 0.2 °,24.47 ° ± 0.2 °,24.95 ° ± 0.2 °,25.42 ° ± 0.2 °,26.51 ° ± 0.2 °, 35 ° ± 0.2 °,30.04 ° ± 0.2 °,32.10 ° ± 0.2 °,32.83 ° ± 0.2 °,33.44 ° ± 0.44 ° ± 0.2 °.

In some embodiments, the phosphate salt of the present invention is phosphate form a, which has an X-ray powder diffraction pattern substantially as shown in fig. 1.

In some embodiments, the phosphate salt of the present invention is phosphate form a, which comprises an endothermic peak at 155.79 ℃ ± 3 ℃.

In some embodiments, the phosphate salt of the present invention is phosphate form a, which has a differential scanning calorimeter substantially as shown in fig. 5.

In some embodiments, the salt of the compound of formula (I) of the present invention is the hydrochloride salt.

In some embodiments, the hydrochloride salt of the present invention is hydrochloride form a, which has an X-ray powder diffraction pattern with diffraction peaks at the following 2θ angles: 7.54 ° ± 0.2 °,12.49 ° ± 0.2 °,15.16 ° ± 0.2 °,16.49 ° ± 0.2 °,18.32 ° ± 0.2 °,19.34 ° ± 0.2 °,19.92 ° ± 0.2 °,22.50 ° ± 0.2 °.

In some embodiments, the hydrochloride salt of the present invention is hydrochloride form a, which has an X-ray powder diffraction pattern with diffraction peaks at the following 2θ angles: 7.05°±0.2°,7.54°±0.2°,12.49°±0.2°,14.13°±0.2°,15.16°±0.2°,16.49°±0.2°,18.32°±0.2°,19.34°±0.2°,19.92°±0.2°,21.28°±0.2°,22.50°±0.2°,23.63°±0.2°,25.40°±0.2°,25.86°±0.2°,26.93°±0.2°,27.42 °±0.2°,27.92 °±0.2°,29.21°±0.2°,29.38°±0.2°,35.14 ++0.2°.

In some embodiments, the hydrochloride salt of the present invention is hydrochloride salt form a having an X-ray powder diffraction pattern substantially as shown in fig. 2.

In some embodiments, the hydrochloride salt of the present invention is hydrochloride form a, which comprises an endothermic peak at 264.34 ℃ ± 3 ℃.

In some embodiments, the hydrochloride salt of the present invention is hydrochloride form a having a differential scanning calorimeter substantially as shown in fig. 6.

In some embodiments, the salt of the compound of formula (I) of the present invention is a maleate salt.

In some embodiments, the maleate salt of the present invention is maleate salt form a, and the X-ray powder diffraction pattern of maleate salt form a has diffraction peaks at the following 2θ angles: 6.25 ° ± 0.2 °,9.37 ° ± 0.2 °,9.66 ° ± 0.2 °,15.08 ° ± 0.2 °,16.60 ° ± 0.2 °,18.93 ° ± 0.2 °,21.06 ° ± 0.2 °,22.95 ° ± 0.2 °,27.99 ° ± 0.2 °.

In some embodiments, the maleate salt of the present invention is maleate salt form a, and the X-ray powder diffraction pattern of maleate salt form a has diffraction peaks at the following 2θ angles: 6.25 ° ± 0.2 °,9.37 ° ± 0.2 °,9.66 ° ± 0.2 °,15.08 ° ± 0.2 °,16.60 ° ± 0.2 °,17.74 ° ± 0.2 °,18.41 ° ± 0.2 °,18.93 ° ± 0.2 °,20.54 ° ± 0.2 °,21.06 ° ± 0.2 °,21.79 ° ± 0.2 °,22.95 ° ± 0.2 °,24.72 ° ± 0.2 °,25.38 ° ± 0.2 °,27.28 ° ± 0.2 °,27.99 ° ± 0.2 °,29.40 ° ± 0.2 °,30.07 ° ± 0.2 °,31.85 ° ± 0.2 °,33.73 ° ± 0.2 °,34.64 ° ± 0.2 °.

In some embodiments, the maleate salt of the present invention is maleate salt form a having an X-ray powder diffraction pattern substantially as shown in figure 3.

In some embodiments, the maleate salt of the present invention is maleate salt form a, and the differential scanning calorimetry trace of maleate salt form a comprises an endothermic peak at 218.14 ℃ ± 3 ℃.

In some embodiments, the maleate salt of the present invention is maleate salt form a having a differential scanning calorimeter substantially as shown in fig. 7.

In some embodiments, the salt of the compound of formula (I) of the present invention is p-toluenesulfonate.

In some embodiments, the p-toluenesulfonate salt of the present invention is p-toluenesulfonate salt form a, the X-ray powder diffraction pattern of which has diffraction peaks at the following 2θ angles: 4.25 ° ± 0.2 °,8.44 ° ± 0.2 °,9.35 ° ± 0.2 °,9.59 ° ± 0.2 °,13.26 ° ± 0.2 °,13.55 ° ± 0.2 °,15.10 ° ± 0.2 °,15.49 ° ± 0.2 °,17.75 ° ± 0.2 °,18.24 ° ± 0.2 °,18.85 ° ± 0.2 °,19.17 ° ± 0.2 °,25.75 ° ± 0.2 °,25.96 ° ± 0.2 °.

In some embodiments, the p-toluenesulfonate salt of the present invention is p-toluenesulfonate salt form a, the X-ray powder diffraction pattern of which has diffraction peaks at the following 2θ angles: 4.25 ° ± 0.2 °,8.44 ° ± 0.2 °,9.35 ° ± 0.2 °,9.59 ° ± 0.2 °,13.26 ° ± 0.2 °,13.55 ° ± 0.2 °,15.10 ° ± 0.2 °,15.49 ° ± 0.2 °,17.75 ° ± 0.2 °,18.24 ° ± 0.2 °,18.85 ° ± 0.2 °,19.17 ° ± 0.2 °,21.00 ° ± 0.2 °,21.15 ° ± 0.2 °,25.75 ° ± 0.2 °,25.96 ° ± 0.2 °.

In some embodiments, the p-toluenesulfonate salt of the present invention is p-toluenesulfonate salt form a, the X-ray powder diffraction pattern of which has diffraction peaks at the following 2θ angles: 4.25 ° ± 0.2 °,8.44 ° ± 0.2 °,9.35 ° ± 0.2 °,9.59 ° ± 0.2 °,11.84 ° ± 0.2 °,13.26 ° ± 0.2 °,13.55 ° ± 0.2 °,15.10 ° ± 0.2 °,15.49 ° ± 0.2 °,16.63 ° ± 0.2 °,17.75 ° ± 0.2 °,18.24 ° ± 0.2 °,18.85 ° ± 0.2 °,19.17 ° ± 0.2 °,19.59 ° ± 0.2 °,21.00 ° ± 0.2 °,21.15 ° ± 0.2 °,21.43 ° ± 0.2 °,22.83 ° ± 0.2 °,23.20 ° ± 0.2 °,24.63 ° ± 0.2 °,25.46 ° ± 0.2 °,25.75 ° ± 0.96 ° ± 0.2 °,28.31 ° ± 0.2 °,29.06 ° ± 0.34 ° ± 0.30 °.

In some embodiments, the p-toluenesulfonate salt of the present invention is p-toluenesulfonate salt form a having an X-ray powder diffraction pattern substantially as shown in fig. 4.

In some embodiments, the p-toluenesulfonate salt of the present invention is p-toluenesulfonate salt form a, the differential scanning calorimetry pattern of p-toluenesulfonate salt form a comprising endothermic peaks at 130.74 ℃ ± 3 ℃ and 242.83 ℃ ± 3 ℃.

In some embodiments, the p-toluenesulfonate salt of the present invention is p-toluenesulfonate salt form a having a differential scanning calorimeter substantially as shown in fig. 8.

In another aspect, the present invention relates to a pharmaceutical composition comprising any of the salts described herein, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, or combination thereof.

In one aspect, the invention relates to the use of said salt or said pharmaceutical composition for the preparation of a medicament for the prevention, treatment or alleviation of a disease state mediated through MAO-B.

In some of these regimens, the disease modulated by MAO-B of the invention is a neurodegenerative disease, a psychotic disorder, or a cancer.

In some of these regimens, the neurodegenerative disease of the invention is parkinson's disease, cerebral ischemia, alzheimer's disease, amyotrophic lateral sclerosis, bovine spongiform encephalopathy, huntington's disease, creutzfeldt-jakob disease, ataxia telangiectasia, cerebellar atrophy, spinal muscular atrophy, primary lateral sclerosis, or multiple sclerosis.

In another aspect, the invention relates to the use of said salt or said pharmaceutical composition for the preparation of a medicament for inhibiting MAO-B.

In another aspect, the present invention also relates to a process for the preparation of a salt of a compound of formula (I) or a crystalline form thereof.

The solvent used in the process for producing a salt or a crystal form thereof of the present invention is not particularly limited, and any solvent which can dissolve the starting material to a degree and does not affect the properties thereof is included in the present invention. In addition, many similar modifications, equivalent substitutions, or equivalent solvents, combinations of solvents, and different proportions of solvent combinations described herein are considered to be encompassed by the present invention. The present invention gives the preferred solvents to be used in each reaction step.

The experiments for preparing the salts or crystal forms thereof according to the present invention will be described in detail in the examples section. Meanwhile, the invention provides pharmacological property test experiments (such as pharmacokinetic experiments), solubility experiments, stability experiments, hygroscopicity experiments and the like of the salt or the crystal form thereof. Experiments prove that the phosphate crystal form A has unexpected technical advantages:

1. the phosphate crystal form A has good stability, for example, no or almost no hygroscopicity, does not change when placed at normal temperature, is stable under high temperature, high humidity and illumination conditions, and has basically no change in appearance and purity; the water solubility is good.

2. The phosphate form a has higher plasma concentration and exposure than other salts, such as hydrochloride form a, maleate form a, p-toluenesulfonate form a, in rats, and thus has better pharmacokinetic properties.

Therefore, the phosphate crystal form A has better bioactivity and higher stability, and is more suitable for pharmaceutical use.

Definitions and general terms

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference in their entirety. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are described herein.

"crystalline form" or "crystalline form" refers to a solid having a highly regular chemical structure, including, but not limited to, single or multicomponent crystals, and/or polymorphs, solvates, hydrates, clathrates, co-crystals, salts, solvates of salts, hydrates of salts of the compounds. The crystalline form of a substance may be obtained by a number of methods known in the art. Such methods include, but are not limited to, melt crystallization, melt cooling, solvent crystallization, crystallization in a defined space, e.g., in a nanopore or capillary, crystallization on a surface or template, e.g., on a polymer, crystallization in the presence of additives such as co-crystallizing anti-molecules, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, reactive crystallization, anti-solvent addition, milling, solvent drop milling, and the like.

"amorphous" or "amorphous form" refers to a substance that forms when particles (molecules, atoms, ions) of the substance are non-periodically arranged in three dimensions, characterized by a diffuse, non-spiking X-ray powder diffraction pattern. Amorphous is a special physical form of solid material whose locally ordered structural features suggest a myriad of interactions with crystalline material. Amorphous forms of a substance can be obtained by a number of methods known in the art. Such methods include, but are not limited to, quenching, antisolvent flocculation, ball milling, spray drying, freeze drying, wet granulation, and solid dispersion techniques, among others.

"solvent" refers to a substance (typically a liquid) that is capable of completely or partially dissolving another substance (typically a solid). Solvents useful in the practice of the present invention include, but are not limited to, water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, methylene chloride, dimethyl sulfoxide, 1, 4-dioxane, ethanol, ethyl acetate, butanol, tertiary butanol, N-dimethylacetamide, N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, mixtures thereof, and the like.

"antisolvent" refers to a fluid that facilitates precipitation of a product (or product precursor) from a solvent. The antisolvent may comprise a cold gas, or a fluid that promotes precipitation by a chemical reaction, or a fluid that reduces the solubility of the product in the solvent; it may be the same liquid as the solvent but at a different temperature, or it may be a different liquid than the solvent.

"solvate" means a compound having a solvent on, in, or on and in the crystal lattice that can be water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, methylene chloride, dimethyl sulfoxide, 1, 4-dioxane, ethanol, ethyl acetate, butanol, t-butanol, N-dimethylacetamide, N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, methyl pyrrolidone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, mixtures thereof, and the like. A specific example of a solvate is a hydrate, wherein the solvent on the surface, in the crystal lattice or both is water. The hydrate may or may not have other solvents than water on the surface of the substance, in the crystal lattice, or both.

The crystalline forms may be identified by a variety of techniques such as X-ray powder diffraction (XRPD), infrared absorption spectroscopy (IR), melting point, differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA), nuclear magnetic resonance, raman spectroscopy, X-ray single crystal diffraction, dissolution calorimetry, scanning Electron Microscopy (SEM), quantitative analysis, solubility and dissolution rate, and the like.

The X-ray powder diffraction (XRPD) can detect the information of crystal form change, crystallinity, crystal structure state and the like, and is a common means for identifying the crystal form. The peak positions of the XRPD patterns are largely dependent on the structure of the crystalline form, relatively insensitive to experimental details, and their relative peak heights depend on many factors related to sample preparation and instrument geometry. Thus, in some embodiments, the crystalline forms of the invention are characterized by XRPD patterns having certain peak positions, substantially as shown in the XRPD patterns provided in the figures of the invention. Meanwhile, the measure of 2θ of the XRPD pattern may have experimental errors, and the measure of 2θ of the XRPD pattern may slightly differ from instrument to instrument and sample to sample, so the value of 2θ cannot be regarded as absolute. Depending on the instrument conditions used in this test, diffraction peaks have a margin of error of + -0.2 deg..

Differential Scanning Calorimeter (DSC) is a method for measuring the temperature of a sample and an inert reference substance (commonly used alpha-Al) by continuously heating or cooling under the control of a program ₂ O ₃ ) A technique in which the energy difference between them varies with temperature. The endothermic peak height of the DSC curve depends on many factors related to sample preparation and instrument geometry, while peak position is relatively insensitive to experimental details. Thus, in some embodiments, the crystalline forms of the invention are characterized by a DSC plot having characteristic peak positions,substantially as shown in the DSC profile provided in the accompanying drawings of the present invention. Meanwhile, the DSC profile may have experimental errors, and the peak position and peak value of the DSC profile may slightly differ from instrument to instrument and from sample to sample, so that the peak position or the value of the DSC endothermic peak cannot be regarded as absolute. Depending on the instrument conditions used in this test, there is an error margin of + -3deg.C for the endothermic peak.

Thermogravimetric analysis (TGA) is a technique for measuring the mass of a substance as a function of temperature under program control, and is suitable for examining the loss of a solvent in a crystal or the sublimation and decomposition processes of a sample, and can be used to infer the presence of water of crystallization or a crystallization solvent in the crystal. The quality change exhibited by the TGA profile depends on many factors such as sample preparation and instrumentation; the quality of TGA detection varies slightly from instrument to instrument and from sample to sample. Depending on the instrument conditions used in this test, there was a margin of error of + -0.1% for the mass change.

In the context of the present invention, the 2 theta values in the X-ray powder diffraction pattern are all in degrees (°).

The term "substantially as shown in the figures" means that at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 99% of the peaks in the X-ray powder diffraction pattern or DSC pattern or raman spectrum or infrared spectrum are shown in the figures.

When referring to a spectrogram or/and data appearing in the graph, a "peak" refers to a feature that one skilled in the art can recognize that is not attributable to background noise.

The present invention relates to salts of said (2 s,4 s) -4-fluoro-1- (4- (3-fluorobenzyloxy) benzyl) pyrrolidine-2-carboxamide and/or crystalline forms thereof, which are present in substantially pure crystalline form.

By "substantially pure" is meant that one form is substantially free of the other form or forms, i.e., the purity of the form is at least 80%, or at least 85%, or at least 90%, or at least 93%, or at least 95%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9%, or the form contains less than 20%, or less than 10%, or less than 5%, or less than 3%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01% of the total volume or total weight of the forms.

By "substantially free" is meant that the percentage of one or more other crystalline forms in the total volume or weight of the crystalline forms is less than 20%, or less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01%.

"relative intensity" (or "relative peak height") in an XRPD pattern refers to the ratio of the intensity of the first intensity peak to the intensity of the first intensity peak of all diffraction peaks of the X-ray powder diffraction pattern, taken as 100%.

In the context of the present invention, when used or whether or not the word "about" or "about" is used, means within 10%, suitably within 5%, particularly within 1% of a given value or range. Alternatively, the term "about" or "approximately" means within an acceptable standard error of the average value to one of ordinary skill in the art. Whenever a number is disclosed having a value of N, any number within the values of N+/-1%, N+/-2%, N+/-3%, N+/-5%, N+/-7%, N+/-8% or N+/-10% will be explicitly disclosed, where "+/-" means plus or minus.

"room temperature" in the present invention means a temperature from about 10℃to about 40 ℃. In some embodiments, "room temperature" refers to a temperature from about 20 ℃ to about 30 ℃; in other embodiments, "room temperature" refers to 20 ℃,22.5 ℃,25 ℃,27.5 ℃, and so forth.

Pharmaceutical compositions, formulations, administration and uses of the salts or crystalline forms thereof of the invention

The pharmaceutical composition of the invention is characterized by comprising salts of the compound shown in the formula (I) and/or crystal forms thereof and pharmaceutically acceptable carriers, auxiliary agents or excipients. The amount of the salt of the compound or crystalline form thereof in the pharmaceutical composition of the invention is effective to detectably treat or ameliorate central nervous system dysfunction in a patient. The pharmaceutical compositions of the present invention may optionally further comprise other therapeutic and/or prophylactic ingredients.

Suitable carriers, adjuvants and excipients are well known to those skilled in the art and are described in detail in, for example, ansel h.c. et al, ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems (2004) Lippincott, williams & Wilkins, philiadelphia; gennaro a.r.et al, remington: the Science and Practice of Pharmacy (2000) Lippincott, williams & Wilkins, philadelphia; and Rowe R.C., handbook of Pharmaceutical Excipients (2005) Pharmaceutical Press, chicago.

The skilled artisan will know and be familiar with the art to which they will be able to select the appropriate amount of suitable pharmaceutically acceptable excipients for use in the present invention. Furthermore, there are a number of resources available to the skilled person, who describe pharmaceutically acceptable excipients and are used to select the appropriate pharmaceutically acceptable excipient. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), the Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

Various carriers for formulating pharmaceutically acceptable compositions, and well known techniques for their preparation, are disclosed in Remington, the Science and Practice of Pharmacy,21st edition,2005,ed.D.B.Troy,Lippincott Williams&Wilkins,Philadelphia,and Encyclopedia of Pharmaceutical Technology,eds.J.Swarbrick and J.C.Boylan,1988-1999,Marcel Dekker,New York, the contents of each of which are incorporated herein by reference. It is within the scope of the present invention to contemplate its use in addition to any common carrier that is incompatible with the compounds of the present invention, such as by producing any undesirable biological effect, or by interacting in a deleterious manner with any other component of the pharmaceutically acceptable composition.

The pharmaceutical compositions of the present invention are prepared using techniques and methods known to those skilled in the art. Some methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

In another aspect, the present invention relates to a process for preparing a pharmaceutical composition comprising a salt of a compound of the invention or a crystalline form thereof and a pharmaceutically acceptable excipient, carrier, adjuvant, vehicle, or combination thereof, which process comprises mixing the various ingredients. Pharmaceutical compositions comprising salts of the compounds of the invention or crystalline forms thereof may be prepared by mixing, for example, at ambient temperature and atmospheric pressure.

Salts of the compounds of the invention, or crystalline forms thereof, are typically formulated into dosage forms suitable for administration to a patient by the desired route. For example, dosage forms include those suitable for the following routes of administration: (1) Oral administration, such as tablets, capsules, caplets, pills, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets and cachets; (2) Parenteral administration, such as sterile solutions, suspensions, and reconstituted powders; (3) transdermal administration, such as transdermal patch tablets; (4) rectal administration, such as suppositories; (5) inhalations, such as aerosols, solutions and dry powders; and (6) topical administration, such as creams, ointments, lotions, solutions, pastes, sprays, foams and gels.

The pharmaceutical composition provided by the invention can be provided in a soft capsule or a hard capsule, and can be prepared from gelatin, methylcellulose, starch or calcium alginate. Hard gelatin capsules, also known as Dry Filled Capsules (DFCs), consist of two segments, one being filled into the other, thus completely encapsulating the active ingredient. Soft Elastic Capsules (SEC) are soft, spherical shells, such as gelatin shells, which are plasticized by the addition of glycerol, sorbitol or similar polyols. The soft gelatin shell may contain a preservative to prevent microbial growth. Suitable preservatives are those described herein, including methyl and propyl parabens, and sorbic acid. Liquid, semi-solid and solid dosage forms provided herein may be encapsulated in capsules. Suitable liquid and semi-solid dosage forms include solutions and suspensions in propylene carbonate, vegetable oils or triglycerides. Capsules containing such solutions can be prepared as described in U.S. patent nos.4,328,245;4,409,239 and 4,410,545. The capsules may also be coated as known to those skilled in the art to improve or maintain dissolution of the active ingredient.

In one embodiment, the methods of treatment of the present invention comprise administering to a patient in need thereof a safe and effective amount of a salt of a compound of the present invention or a crystalline form thereof or a pharmaceutical composition comprising a salt of a compound of the present invention or a crystalline form thereof. Embodiments of the present invention include treating the diseases mentioned herein by administering to a patient in need thereof a safe and effective amount of a salt of the compound of the present invention or a crystalline form thereof or a pharmaceutical composition comprising the salt of the compound of the present invention or a crystalline form thereof.

In one embodiment, the salts of the compounds of the invention or crystalline forms thereof or pharmaceutical compositions comprising the salts of the compounds of the invention or crystalline forms thereof may be administered by any suitable route of administration, including systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, and rectal administration. Typical parenteral administration refers to administration by injection or infusion and includes intravenous, intramuscular, and subcutaneous injection or infusion. Topical administration includes application to the skin, intraocular, otic, intravaginal, inhalation, and intranasal administration. In one embodiment, the salt of the compound of the invention or a crystalline form thereof or a pharmaceutical composition comprising the salt of the compound of the invention or a crystalline form thereof may be administered orally. In another embodiment, a salt of the compound of the invention or a crystalline form thereof or a pharmaceutical composition comprising a salt of the compound of the invention or a crystalline form thereof may be administered by inhalation. In yet another embodiment, a salt of a compound of the invention or a crystalline form thereof or a pharmaceutical composition comprising a salt of a compound of the invention or a crystalline form thereof may be administered intranasally.

In one embodiment, a salt of a compound of the invention or a crystalline form thereof or a pharmaceutical composition comprising a salt of a compound of the invention or a crystalline form thereof may be administered at one time or, depending on the dosing regimen, several times at different time intervals over a specified period of time. For example, once, twice, three times or four times daily. In one embodiment, the administration is once daily. In yet another embodiment, the administration is twice daily. The administration may be performed until the desired therapeutic effect is achieved or the desired therapeutic effect is maintained indefinitely. Suitable dosing regimens for salts of the compounds of the invention or crystalline forms thereof or pharmaceutical compositions comprising the salts of the compounds of the invention or crystalline forms thereof depend on the pharmacokinetic properties of the salts of the compounds, such as absorption, distribution and half-life, which can be determined by the skilled person. Furthermore, suitable dosing regimens for salts of the compounds of the invention, or crystalline forms thereof, or pharmaceutical compositions comprising the salts of the compounds of the invention, or crystalline forms thereof, include factors within the knowledge and experience of the skilled artisan, including the duration of time for which the regimen is performed, the disease to be treated, the severity of the disease to be treated, the age and physical condition of the patient to be treated, the medical history of the patient to be treated, the nature of concurrent therapy, the desired therapeutic effect, and the like. Such a skilled artisan will also appreciate that adjustments to the appropriate dosing regimen may be required for the individual patient's response to the dosing regimen, or as the individual patient needs to change over time.

The salts of the compounds of the invention or crystalline forms thereof may be administered simultaneously with, or before or after, one or more other therapeutic agents. The salts of the compounds of the present invention or crystalline forms thereof may be administered separately from other therapeutic agents by the same or different routes of administration, or in the same pharmaceutical composition as they are.

The term "therapeutically effective amount" as used herein refers to the total amount of each active ingredient sufficient to exhibit a beneficial therapeutic effect. For example, an amount sufficient to treat, cure, or alleviate symptoms of the disease is administered or equilibrated in vivo. The effective amount required for a particular therapeutic regimen will depend upon a variety of factors including the disease being treated, the severity of the disease, the activity of the particular agent being used, the mode of administration, the rate of clearance of the particular agent, the duration of the treatment, the combination, the age, body weight, sex, diet and health of the patient, etc. Other factors that need to be considered in the art for "therapeutically effective amounts" can be described in Gilman et al, eds., goodman And Gilman's: the Pharmacological Bases of Therapeutics,8th ed., pergamon Press,1990; remington's Pharmaceutical Sciences,17th ed., mack Publishing Company, easton, pa.,1990. The compounds of the present invention have in vivo activity when administered orally, intraperitoneally, or intravenously in a therapeutically effective amount of 0.1-200 mg/kg.

The optimal therapeutically effective amount to be administered can be readily determined by one skilled in the art and will vary substantially depending on the strength of the formulation, the mode of administration, and the advancement of the disease or condition being treated. In addition, factors related to the particular subject being treated include the subject's age, weight, diet, and time of administration, resulting in the need to adjust the dosage to an appropriate therapeutically effective level.

The term "administering" refers to providing a therapeutically effective amount of a drug to an individual by means including oral, sublingual, intravenous, subcutaneous, transdermal, intramuscular, intradermal, intrathecal, epidural, intraocular, intracranial, inhalation, rectal, vaginal, and the like. The administration form includes paste, lotion, tablet, capsule, pill, powder, granule, suppository, pellet, lozenge, injection, sterile solution or nonaqueous solution, suspension, emulsion, patch, etc. The active ingredient is compounded with a non-toxic pharmaceutically acceptable carrier (e.g., dextrose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, silica gel, potato starch, urea, dextran, etc.).

The preferred route of administration will vary with clinical characteristics, and the dosage will vary depending on the condition of the patient being treated, and the physician will determine the appropriate dosage for the individual patient. The therapeutically effective amount per unit dose depends on the body weight, physiology and the chosen vaccination regimen. The weight of the compound per unit dose is the weight of the compound per administration and does not include the weight of the carrier (the carrier is contained in the drug). Pharmaceutical compositions comprising a compound of formula (I) as defined above comprise from about 0.1mg to about 500mg, most preferably from 1 to 10mg, of one or more active ingredients per unit dose, e.g., capsule, tablet, powder injection, teaspoon capacity, suppository, etc.

The salt of the compound or the crystal form and the pharmaceutical composition thereof provided by the invention can be used for preparing medicines for preventing, treating or relieving diseases regulated by MAO-B of patients, medicines for preventing, treating or relieving neurodegenerative diseases, psychosis or cancers, and medicines for inhibiting the activity of MAO-B.

In particular, the amount of the compound in the pharmaceutical composition of the present invention is effective to detectably and selectively inhibit the activity of MAO-B, and the salt of the compound of the present invention or a crystalline form thereof can be used as a medicament for treating diseases modulated by MAO-B, such as Parkinson's disease.

The salts of the compounds of the present invention or crystalline forms thereof may be used, but are in no way limited to, the use of an effective amount of a salt of the compound of the present invention or a crystalline form or pharmaceutical composition thereof for administration to a patient to prevent, treat or ameliorate neurodegenerative diseases. The neurodegenerative disease further includes, but is not limited to, parkinson's disease, cerebral ischemia, alzheimer's disease, amyotrophic lateral sclerosis, hearing loss due to aging, dementia, retinal degeneration, macular degeneration, glaucoma, bovine spongiform encephalopathy, huntington's chorea, creutzfeldt-jakob disease, ataxia telangiectasia, cerebellar atrophy, spinal muscular atrophy, primary lateral sclerosis or multiple sclerosis.

Salts of the compounds of the present invention or crystalline forms thereof may be used, but are in no way limited to, administration to a patient of an effective amount of a compound or pharmaceutical composition of the present invention to prevent, treat or alleviate psychosis. The psychosis is schizophrenia and/or anxiety, wherein schizophrenia further includes but is not limited to brief psychotic disorder, delusional disorder, schizoaffective disorder, and schizophreniform disorder; wherein anxiety disorders further include, but are not limited to, panic disorder, obsessive-compulsive disorder, post-traumatic stress disorder, social phobia or social anxiety disorder, specific phobia, and generalized anxiety disorder.

Salts of the compounds of the present invention or crystalline forms thereof may be used, but are in no way limited to, administration to a patient of an effective amount of a compound or pharmaceutical composition of the present invention to prevent, treat or ameliorate cancer. The cancers further include, but are not limited to, prostate cancer, breast cancer, testicular cancer, colorectal cancer, lung cancer, brain tumor, kidney tumor, or blood cancer.

An "effective amount" or "effective dose" of a salt of a compound of the invention or a crystalline form or pharmaceutically acceptable composition thereof refers to an amount effective to treat or reduce the severity of one or more of the conditions referred to herein. According to the methods of the invention, the salts of the compounds of the invention, or crystalline forms or pharmaceutically acceptable compositions thereof, may be in any amount and by any route of administration effective to treat or reduce the severity of the disease. The exact amount necessary will vary depending on the patient's condition, depending on the race, age, general condition of the patient, severity of the infection, particular factors, mode of administration, and the like. Salts of the compounds of the invention, or crystalline forms or pharmaceutically acceptable compositions thereof, may be administered in combination with one or more other therapeutic agents, as discussed herein.

The salts of the compounds of the invention or their crystalline forms and pharmaceutical compositions are useful in veterinary therapy for mammals, in addition to human therapy, in companion animals, animals of introduced species and farm animals. Examples of other animals include horses, dogs, and cats.

Drawings

FIG. 1 is an X-ray powder diffraction (XRPD) pattern of phosphate form A of a compound of formula (I).

Fig. 2 is an X-ray powder diffraction (XRPD) pattern of hydrochloride salt form a of the compound of formula (I).

Fig. 3 is an X-ray powder diffraction (XRPD) pattern of maleate salt form a of the compound of formula (I).

Fig. 4 is an X-ray powder diffraction (XRPD) pattern of para-toluenesulfonate salt form a of the compound of formula (I).

FIG. 5 is a Differential Scanning Calorimeter (DSC) of phosphate form A of a compound of formula (I).

FIG. 6 is a Differential Scanning Calorimeter (DSC) of hydrochloride form A of the compound of formula (I).

FIG. 7 is a Differential Scanning Calorimeter (DSC) of maleate form A of a compound of formula (I).

FIG. 8 is a Differential Scanning Calorimeter (DSC) of the p-toluenesulfonate salt form A of the compound of formula (I).

Detailed Description

The invention is further illustrated by way of examples which are not intended to limit the scope of the invention.

The X-ray powder diffraction analysis method used in the invention comprises the following steps: an Empyrean diffractometer was used to obtain X-ray powder diffraction patterns using Cu-K alpha radiation (45 KV,40 mA). The powdered sample was prepared as a thin layer on a monocrystalline silicon sample holder, placed on a rotating sample stage and analyzed in steps of 0.0167 ° in the range of 3 ° -40 °. Data was collected using Data Collector software, highScore Plus software processed the Data, and Data Viewer software read the Data.

The Differential Scanning Calorimeter (DSC) analysis method used in the invention comprises the following steps: differential scanning calorimeter was performed using a TA Q2000 module with a thermal analysis controller. Data were collected and analyzed using TA Instruments Thermal Solutions software. About 1-5mg of the sample was accurately weighed into a specially made aluminum crucible with a lid, and sample analysis was performed from room temperature to about 300 ℃ using a linear heating device of 10 ℃/min. During use, the DSC cell was purged with dry nitrogen.

The solubility of the invention is measured by an Agilent 1200 high performance liquid chromatograph DAD/VWD detector, and the model of the chromatographic column is Agilent XDB-C18 (4.6X10 mm,5 μm). The detection wavelength is 266nm, the flow rate is 1.0mL/min, the column temperature is 35 ℃, and the mobile phase A: acetonitrile/0.01M ammonium acetate = 10/90 (V/V) analytical method: acetonitrile/mobile phase a=70/30 (V/V), run time: 10 minutes.

Detailed description of the preferred embodiments

A compound of formula (I): (2S, 4S) -4-fluoro-1- (4- (3-fluorobenzyloxy) benzyl) pyrrolidine-2-carboxamide, a specific synthetic method is described in example 1 of International application WO 201917515 A1.

Examples

Example 1: the phosphate crystal form A of the invention

1. Preparation of phosphate form A

The compound (100 mg) represented by the formula (I) was added to ethyl acetate (2.5 mL), followed by addition of phosphoric acid (50 mg, mass fraction: 85%) and reaction at room temperature overnight. Stopping the reaction, filtering and drying to obtain white solid powder which is the phosphate crystal form A.

2. Identification of phosphate form A

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, there are the following characteristic peaks expressed in degrees 2θ:4.28 °,12.13 °,12.22 °,12.83 °,13.07 °,16.29 °,17.06 °,17.77 °,18.14 °,18.80 °,19.39 °,19.81 °,20.05 °,20.34 °,22.70 °,23.15 °,23.91 °,24.47 °,24.95 °,25.42 °,26.51 °,26.72 °,30.04 °,32.10 °,32.83 °,33.44 °,34.72 °, there is an error margin of ±0.2 °.

(2) Identification by TA Q2000 Differential Scanning Calorimeter (DSC) analysis: the scan rate was 10 c/min, including an endothermic peak at 155.79 c, with an error margin of ± 3 c.

Example 2: the hydrochloride of the invention is in the form A

1. Preparation of hydrochloride form A

The compound (100 mg) represented by the formula (I) was added to ethyl acetate (2 mL), followed by addition of hydrochloric acid (40 mg, mass fraction: 36-38%) and reaction at room temperature overnight. Stopping the reaction, filtering, and drying to obtain white solid powder which is hydrochloride crystal form A.

2. Identification of hydrochloride form A

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, there are the following characteristic peaks expressed in degrees 2θ:7.05 °,7.54 °,12.49 °,14.13 °,15.16 °,16.49 °,18.32 °,19.34 °,19.92 °,21.28 °,22.50 °,23.63 °,25.40 °,25.86 °,26.93 °,27.42 °,27.92 °,29.21 °,29.38 °,35.14 °, there is an error margin of ±0.2 °.

(2) Identification by TA Q2000 Differential Scanning Calorimeter (DSC) analysis: the scan rate was 10 c/min, including an endothermic peak at 264.34 c, with an error margin of ± 3 c.

Example 3: the maleate salt of the present invention form A

1. Preparation of maleate salt form A

The compound (100 mg) represented by the formula (I) was added to ethyl acetate (2 mL), followed by maleic acid (41 mg), and the mixture was reacted at room temperature overnight. Stopping the reaction, carrying out suction filtration and drying to obtain white solid powder which is the maleate crystal form A.

2. Identification of maleate salt form A

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, there are the following characteristic peaks expressed in degrees 2θ:6.25 °,9.37 °,9.66 °,15.08 °,16.60 °,17.74 °,18.41 °,18.93 °,20.54 °,21.06 °,21.79 °,22.95 °,24.72 °,25.38 °,27.28 °,27.99 °,29.40 °,30.07 °,31.85 °,33.73 °,34.64 °, there is an error margin of ±0.2 °.

(2) Identification by TA Q2000 Differential Scanning Calorimeter (DSC) analysis: the scan rate was 10 c/min, including an endothermic peak at 218.14 c, with an error margin of ± 3 c.

Example 4: the p-toluenesulfonate crystal form A of the invention

1. Preparation of p-toluenesulfonate form A

The compound (100 mg) represented by the formula (I) was added to ethyl acetate (3 mL), and p-toluenesulfonic acid (71 mg) was added thereto, and the mixture was reacted at room temperature overnight. Stopping the reaction, filtering, and drying to obtain white solid powder which is p-toluenesulfonate crystal form A.

2. Identification of para-toluenesulfonate form A

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, there are the following characteristic peaks expressed in degrees 2θ:4.25 °,8.44 °,9.35 °,9.59 °,11.84 °,13.26 °,13.55 °,15.10 °,15.49 °,16.63 °,17.75 °,18.24 °,18.85 °,19.17 °,19.59 °,21.00 °,21.15 °,21.43 °,22.83 °,23.20 °,24.63 °,25.46 °,25.75 °,25.96 °,28.31 °,29.06 °,30.34 °, there is an error margin of ±0.2 °.

(2) Identification by TA Q2000 Differential Scanning Calorimeter (DSC) analysis: the scan rate was 10 ℃/min, including endothermic peaks at 130.74 ℃ and 242.83 ℃, with a margin of error of + -3 ℃.

EXAMPLE 5 pharmacokinetic experiments on the salts or crystalline forms of the invention

The sample for administration (i.e., the salt of the present invention or a crystalline form thereof, or the compound of formula (I) of the present invention) is filled into capsules for oral administration.

Taking 3 male rats of 0.2-0.3kg, orally administering capsule containing test sample at dosage of 5mg/kg, and collecting blood at time point 0.25,0.5,1.0,2.0,5.0,7.0 and 24 hr. A standard curve of a suitable range is established according to the concentration of the sample, the concentration of the sample to be tested in the plasma sample is determined in MRM mode by using AB SCIEX API5500 type LC-MS/MS, and quantitative analysis is carried out. According to the drug concentration-time curve, the pharmacokinetic parameters were calculated using the WinNonLin 6.3 software non-compartmental model method. The experimental results are shown in table 1.

TABLE 1 pharmacokinetic experimental data for the salts or crystalline forms of the invention

Sample for sample	T _max (h)	C _max (ng/ml)	AUC _last (h*ng/ml)
				Example 1 (phosphate Crystal form A)	1.15	471	1020
Example 2 (hydrochloride Crystal form A)	0.7	143	378
				Example 3 (maleate salt form A)	3.4	151	450
Example 4 (para-toluenesulfonate Crystal form A)	2.2	162	470
				Compounds of formula (I)	0.8	256	548

Conclusion of experiment:

as shown in table 1, compared with the compound shown in formula (I), the hydrochloride crystal form a, the maleate crystal form a and the p-toluenesulfonate crystal form a of the compound shown in formula (I), the phosphate crystal form a of the present invention has higher blood concentration and larger exposure in rats and has better pharmacokinetic properties.

EXAMPLE 6 stability test of the salt or Crystal form of the invention

(1)High temperature experiments: taking a batch of samples for sample preparation, putting the samples into a flat weighing bottle, spreading the samples into a thin layer with the thickness less than or equal to 5mm, placing the weighing bottle into a constant temperature box with the temperature of 40+/-2 ℃/75+/-5% RH and 60+/-2 ℃/75+/-5% RH for 10 days, sampling on the 5 th day and the 10 th day, and detecting according to the stability key investigation items: the color change of the sample was observed and the purity of the sample was checked by HPLC.

(2)High humidity experiment: a batch of samples for sample preparation is taken and put into a flat weighing bottle, spread into a thin layer with the thickness less than or equal to 5mm, placed for 10 days under the conditions of 25 ℃ and RH 75% +/-5% or RH 90% +/-5%, sampled on the 5 th and 10 th days, and detected according to stability key investigation projects: the color change of the sample was observed and the purity of the sample was checked by HPLC.

(3)Illumination test: taking a batch of samples, placing into a flat weighing bottle, spreading into a thin layer with thickness less than or equal to 5mm, placing into an illumination box (with ultraviolet lamp), and placing the sample into a light box with illuminance of 4500+ -500 lx and ultraviolet light of more than or equal to 0.7w.h/m ² Is placed for 16 days, is sampled on days 5, 10 and 16, and is tested according to the stability key investigation project: observing the color of the sampleThe sample purity was checked by HPLC.

The experimental results show that the appearance and purity of the phosphate crystal form A are not obviously changed under the conditions of high temperature, high humidity and illumination. Namely, the phosphate crystal form A has better stability under various lofting conditions and is suitable for pharmaceutical use.

EXAMPLE 7 hygroscopicity test of the salt or Crystal form of the invention

1. Experimental method

1) The dried glass weighing bottle with plug (with the outer diameter of 50mm and the height of 15 mm) is placed in a constant temperature dryer (with the lower part placed with ammonium chloride saturated solution) at 25+/-1 ℃ in the previous day, and is precisely weighed (m ₁ )。

2) Taking a proper amount of sample, spreading in the weighing bottle, and precisely weighing (m ₂ )。

3) The weighing bottle is opened and placed under the constant temperature and humidity condition for 24 hours together with the bottle cap.

4) The lid of the weighing bottle is covered, and the bottle is precisely weighed (m ₃ ) And (3) calculating: percent weight gain = (m ₃ -m ₂ )/(m ₂ -m ₁ )×100％

5) The wet permeability results are shown in Table 2.

TABLE 2 determination of moisture wicking results

From the experimental results, the phosphate crystal form A disclosed by the invention has no or almost no hygroscopicity and is not easy to be deliquesced due to the influence of high humidity.

EXAMPLE 8 solubility testing of the salts or crystalline forms of the invention

And (3) placing the sample in organic ultrapure water at 37 ℃ to prepare supersaturated solution, oscillating for 24 hours, filtering with a water-based filter membrane to obtain filtrate, and detecting the solubility of the target sample in water by using an HPLC method. The experimental results are shown in table 3.

TABLE 3 solubility test data for the salts or crystalline forms of the invention

Sample for sample	Concentration of the Compound of formula (I) in saturated aqueous solution (. Mu.g/mL)
		Example 1 (phosphate Crystal form A)	19493.14
Example 2 (hydrochloride Crystal form A)	4719.01
		Example 3 (maleate salt form A)	575.35
Example 4 (para-toluenesulfonate Crystal form A)	1571.18
		Compounds of formula (I)	11.40

Conclusion of experiment:

experimental results show that compared with the compound shown in the formula (I), the hydrochloride crystal form A, the maleate crystal form A and the p-toluenesulfonate crystal form A of the compound shown in the formula (I), the phosphate crystal form A has higher solubility in water, so that the phosphate crystal form A has better drug property and is suitable for formulation development.

The above description is merely a basic description of the inventive concept, and any equivalent transformation according to the technical solution of the present invention shall fall within the protection scope of the present invention.

In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.

While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.

Claims

1. Salts of the compounds of formula (I),

(I)，

the method is characterized in that the salt is phosphate; wherein the phosphate is a phosphate form a, and an X-ray powder diffraction pattern of the phosphate form a has diffraction peaks at the following 2θ angles: 4.28 ° ± 0.2 °, 12.13 ° ± 0.2 °, 12.22 ° ± 0.2 °, 12.83 ° ± 0.2 °, 13.07 ° ± 0.2 °, 16.29 ° ± 0.2 °, 17.06 ° ± 0.2 °, 17.77 ° ± 0.2 °, 18.14 ° ± 0.2 °, 18.80 ° ± 0.2 °, 19.39 ° ± 0.2 °, 19.81 ° ± 0.2 °, 20.05 ° ± 0.2 °, 20.34 ° ± 0.2 °, 23.15 ° ± 0.2 °, 25.42 ° ± 0.2 ° 26.72 ° ± 0.2 °.

2. The salt of claim 1, wherein the phosphate salt is phosphate form a, and the X-ray powder diffraction pattern of the phosphate form a has diffraction peaks at the following 2Θ angles: 4.28 ° ± 0.2 °, 12.13 ° ± 0.2 °, 12.22 ° ± 0.2 °, 12.83 ° ± 0.2 °, 13.07 ° ± 0.2 °, 16.29 ° ± 0.2 °, 17.06 ° ± 0.2 °, 17.77 ° ± 0.2 °, 18.14 ° ± 0.2 °, 18.80 ° ± 0.2 °, 19.39 ° ± 0.2 °, 19.81 ° ± 0.2 °, 20.05 ° ± 0.2 °, 20.34 ° ± 0.2 °, 22.70 ° ± 0.2 °, 23.15 ° ± 0.2 °, 23.91 ° ± 0.2 °, 24.47 ° ± 0.2 °, 24.95 ° ± 0.2 °, 25.42 ° ± 0.2 °, 26.51 ° ± 0.2 °, 35 ° ± 0.2 °, 30.04 ° ± 0.2 °, 32.10 ° ± 0.2 °, 32.83 ° ± 0.2 °, 33.44 ° ± 0.44 ° ± 0.2 °.

3. The salt according to any one of claims 1-2, wherein the phosphate salt is phosphate form a, the phosphate form a having an X-ray powder diffraction pattern substantially as shown in figure 1.

4. The salt of claim 1, wherein the phosphate salt is phosphate form a, and the differential scanning calorimetry trace of phosphate form a comprises an endothermic peak at 155.79 ± 3 ℃.

5. The salt of claim 4, wherein the phosphate salt is phosphate form a having a differential scanning calorimetry pattern substantially as shown in figure 5.

6. A pharmaceutical composition comprising the salt of any one of claims 1-5, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, or combination thereof.

7. Use of a salt according to any one of claims 1-5 or a pharmaceutical composition according to claim 6 for the manufacture of a medicament for preventing, treating or alleviating a disease mediated by MAO-B in a patient;

wherein the disease modulated by MAO-B is a neurodegenerative disease, a psychotic disorder or a cancer;

wherein the neurodegenerative disease is Parkinson's disease, cerebral ischemia, alzheimer's disease, amyotrophic lateral sclerosis, bovine spongiform encephalopathy, huntington's chorea, creutzfeldt-Jakob disease, ataxia telangiectasia, cerebellar atrophy, spinal muscular atrophy, primary lateral sclerosis or multiple sclerosis.