Patent Description:
Pain is divided into inflammatory pain, neuropathic pain, nociceptive pain, psychotic (psychogenic) pain, etc., depending on the cause. The inflammatory pain is a pain generated as going with inflammation caused by extracorporeal and nociceptive mechanical stimulation, heat stimulation, chemical stimulation, and the like. It is known that when an inflammatory pain occurs, not only inflammatory sites, but also inflammatory cytokines and cyclooxygenase existed in the spinal cord also play important roles. Neuropathic pain is a pathological pain caused by dysfunction of the peripheral or central nervous system itself. The psychotic pain is a pain generated due to an injury of a normal tissue or the application of a nociceptive stimulation which possibly causes an injury of a normal tissue, and is divided into bodily pain and visceral pain.

As a therapeutic agent for inflammatory pain, a cyclooxygenase (COX) inhibitor such as indomethacin, a cyclooxygenase II (COX-II) inhibitor such as celecoxib, a central analgesic such as tramadol, an analgesic-antipyretic agent such as paracetamol, and the like may be used. However, if the cyclooxygenase inhibitor is used for a prolonged period, as a side effect sometimes gastrointestinal disturbance may be caused, which is problematic. Additionally, as reported the cyclooxygenase II inhibitor may also cause gastric ulcer, and recently its side effects in heart circulation system, such as myocardial infarction, cerebral infarction and the like, also come as problem issues.

As a therapeutic agent for neuropathic pain, a papaverine-like analgesic such as morphine, an anticonvulsive drug such as gabapentin and pregabalin may be used, but it is known that along with the prolonged use time, it is sometimes necessary to increase the dosage and thus a side effect such as sedation may be caused. Currently there are no existing pharmaceutical agent which can be administrated safely without any side effect.

The patent literature <NUM> (<CIT>) described that <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol is capable of selectively inhibiting adenylate cyclase <NUM> and can be used for treating neuropathic pain and/or inflammatory pain. However, the patent literature <NUM> did provide neither a method suitable for preparing <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol for industrial application, nor a crystal form suitable for pharmaceutical preparation. Moreover, in respect of residual organic solvent in the final product, in different preparation and crystallization methods it is desirable to avoid the use of two or more types of organic solvents harmful to mammals, thereby reducing the adverse effect of residual organic solvent on the user.

For the polymorphic form of a medicament, different polymorphic forms may have different chemical and physical properties, including the melting point, chemical stability, apparent solubility, dissolution rate, optical and mechanical properties, vapor pressure, density and the like. These properties may directly or indirectly affect the processing or production of bulk pharmaceutical chemicals and formulations, and may also affect the stability, solubility and bioavailability of the formulations. Thus, the polymorphic form of the medicament is important for the quality, safety and efficacy of the pharmaceutical formulation. For <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol, the requirement thereof in the art is: having a polymorphic form which is suitable for industrial production and has excellent physicochemical properties.

The polymorphism in pharmaceuticals is one of essential factors which affect the pharmaceutical quality and clinically therapeutic effect. The patent literature <NUM> (<CIT>) described that the <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol is capable of selectively inhibiting adenylate cyclase <NUM> and can be used for treating neuropathic pain and/or inflammatory pain, but provided no crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol applicable to clinical application.

Therefore, disclosed herein is a crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol for use in a medicament for the treatment of neuropathic pain and/or inflammatory pain, which has excellent performance in efficacy and safety.

Additionally, through extensive studies, the inventor surprisingly finds a synthesis route of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol and obtains a new crystal form, which successfully solves the problems existed in the prior art. The new crystal form has advantages of excellent physicochemical properties, great stability, and being more suitable for industrial preparation.

The present invention provides a method for the preparation of a crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol, which has the following data of powder x ray diffraction:
the crystal is measured by using Cu-Ka and shows a characteristic peak at an interplanar crystal spacing d of <NUM>° ± <NUM>°, <NUM>° ± <NUM>°, <NUM>° ± <NUM>°, <NUM>°±<NUM>°, <NUM>°±<NUM>°, <NUM>° ± <NUM>°, <NUM>° ± <NUM>°, <NUM>° ± <NUM>° and <NUM>° ± <NUM>° in powder x ray diffraction.

Disclosed herein is a synthesis route of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol as below:
<CHM>.

Disclosed herein is a medicament containing the new crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol.

The medicament disclosed herein is an inhibitor of adenylate cyclase <NUM>.

The medicament disclosed herein is an analgesic.

The medicament disclosed herein is a medicament for the treatment of neuropathic pain and/or inflammatory pain.

The medicament disclosed herein is a medicament for the treatment of cancerous pain.

The medicament disclosed herein is a medicament for the treatment of anxiety caused by the neuropathic pain and other disease-related anxiety and depression.

The medicament disclosed herein is a medicament for the treatment of chronic visceral pain and anxiety and depression associated thereto.

Disclosed herein is the use of the crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol in the manufacture of a medicament for the treatment of neuropathic pain and/or inflammatory pain.

Disclosed herein is the use of the crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol in the manufacture of a medicament for the treatment of anxiety caused by the neuropathic pain and other disease-related anxiety and depression.

Disclosed herein is the use of the crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol in the manufacture of a medicament for the treatment of chronic visceral pain and anxiety and depression associated thereto.

Disclosed herein is the use of the crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol in the treatment of neuropathic pain and/or inflammatory pain.

Disclosed herein is the use of the crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol in treatment of anxiety caused by the neuropathic pain and other disease-related anxiety and depression.

Disclosed herein is the use of the crystal of <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol in the treatment of chronic visceral pain and anxiety and depression associated thereto.

The <NUM>-((<NUM>-(<NUM>-amino)-<NUM>H-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol of the present invention has the following structure:
<CHM>.

This compound is numbered as NB001 in the present invention.

The compound NB001 of the present invention can be synthesized and purified through the following scheme.

Compound <NUM> is <NUM>-(<NUM>-Aminopurin-<NUM>-yl)ethanol, a commercially available product from e.g., J&K Scientific, Meryer (Shanghai) Chemical Technology Co. , or INTATRADE GmbH of Germany, and in the embodiments of the present invention the compound <NUM> is purchased from Cool Pharm.

Compound <NUM> is <NUM>-Amino-<NUM>-pentanol, a product available from Wako Pure Chemical Industries, Ltd. or HBCChem, Inc. , and the compound <NUM> of the present invention is purchased from TCI.

A compound <NUM> is prepared through a substitution reaction of the compound <NUM> with thionyl chloride in accordance with the afore-mentioned scheme. The substitution reaction is carried out in a conventional manner or in a solvent which does not affect the reaction.

By way of example, the solvent which does not affect the reaction may include: ethers such as dioxane, tetrahydrofuran, <NUM>,<NUM>-dimethoxyethane and the like; halogenated hydrocarbons such as chloroform; aromatic hydrocarbons such as toluene; amides such as N,N-dimethylformamide; and sulfoxides such as dimethyl sulfoxide and the like. More than two of these solvents may be mixed in an appropriate ratio when used. The amount of these solvents is, for example, <NUM> to <NUM> times by volume of the compound <NUM>.

The reaction temperature is generally about <NUM> to about <NUM>, and preferably <NUM> to <NUM>.

The reaction time is generally about <NUM> to about <NUM> hours.

The compound <NUM> obtained in such a manner can be isolated and purified by a well-known separation and purification method such as concentration, concentration under reduced pressure, solvent extraction, crystallization, recrystallization, phase transfer, chromatography and the like. It should be noted that compound <NUM> can be used directly in subsequent reactions without isolation.

Next, compound <NUM> is subjected to a condensation reaction with compound <NUM> to obtain the compound NB001 of the present invention.

The reaction is carried out in a conventional manner and in a solvent which does not affect the reaction.

By way of example, the solvent which does not affect the reaction may include: hydrocarbons such as hexane and the like; alcohols such as methanol and the like; ethers such as tetrahydrofuran and the like; esters such as ethyl acetate and the like; halogenated hydrocarbons such as chloroform; aromatic hydrocarbons such as toluene; amides such as N,N-dimethylformamide; and sulfoxides such as dimethyl sulfoxide and the like. More than two of these solvents may be mixed in an appropriate ratio when used. The amount of these solvents is, for example, <NUM> to <NUM> times by volume of the compound <NUM>.

The reaction temperature is usually about <NUM> to about <NUM>, and preferably <NUM> to <NUM>.

The compound NB001 obtained in such a manner can be isolated and purified by a well-known separation and purification method such as concentration, concentration under reduced pressure, solvent extraction, crystallization, recrystallization, phase transfer, chromatography and the like. It should be noted that compound NB001 can be used directly in subsequent reactions without isolation.

Examples of the solvate crystals of the compound NB001 include, for example, alcohol solvate crystals such as methanol solvate crystals, ethanol solvate crystals and the like; organic solvent hydrate crystals having water and an organic solvent (e.g., alcohol solvate hydrate crystals, such as methanol hydrate crystallization, ethanol hydrate crystallization, etc.); and the like.

The crystal disclosed herein can be produced by subjecting the amorphous compound NB001, or other crystal forms of the compound NB001, to a crystal transition. The change in crystal form is a phenomenon in which the crystal structure changes when the temperature or pressure exceeds a certain level.

Examples of the crystal transition method include a method known per se, for example, by crystallization of solution (e.g., a concentration method, a slow cooling method, a reaction method (a diffusion method, and an electrolysis method), a hydrothermal growth method, and a flux method), by vapor crystallization (e.g., a gasification process (a pipe sealing method, and an airflow method), a vapor reaction method, and a chemical transport method), by a melt crystallization method (e.g., a conventional freezing method (a lifting method, a temperature gradient method, and a Bridgman method), a zone melting method (a zone leveling method, and a floating zone method), a special growth method (a VLS method, a liquid phase epitaxy method), a stream fog method (wherein the crystal is dissolved in a solvent, filtered, and the solvent is evaporated under atmospheric conditions), a slurry method (wherein the crystal is added to a solvent such that an excess amount of solid is retained therein to obtain a suspension, the suspension is stirred at room temperature or under a heating or cooling condition, and then the solid is collected by filtration), and methods such as drying under reduced pressure, grinding, pulverization, pressurization and the like.

For obtaining the crystal disclosed herein, the slurry method is particularly preferred among the afore-mentioned methods. Specifically, the method is preferably as follows: adding the crystal of the compound NB0011 into a solvent such that an excess amount of solid is retained therein to obtain a suspension, stirring the suspension, and collecting the solid by filtration. The solvent as used includes, for example, aromatic hydrocarbons (e.g., benzene, toluene, xylene, etc.), halogenated hydrocarbons (e.g., dichloromethane, chloroform, etc.), saturated hydrocarbons (e.g., hexane, heptane, cyclohexane, etc.), ethers (e.g., diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, etc.), nitriles (e.g., acetonitrile, etc.), ketones (e.g., acetone, etc.), sulfoxides (e.g., dimethylsulfoxide, etc.), amides (e.g., N,N-dimethylformamide, etc.), esters (e.g., ethyl acetate, etc.), alcohols (e.g., methanol, ethanol, <NUM>-propanol, etc.), water and the like. These solvents may be used alone or in combination of two or more in an appropriate ratio (e.g., <NUM>:<NUM> to <NUM>:<NUM>). The solvent is preferably alcohols (e.g., <NUM>-propanol, etc.), ketones (methyl ethyl ketone, etc.) and esters (e.g., ethyl acetate, etc.), and more preferably ketones (methyl ethyl ketone, etc.).

The amount of the solvent as used is generally about <NUM> to about <NUM>, and preferably about <NUM> to about <NUM>, relative to the crystal of compound NB001 (<NUM>).

The suspension is preferably stirred at room temperature or from about <NUM> to about <NUM>, and more preferably from about <NUM> to about <NUM>. In the present specification, room temperature refers to a temperature of about <NUM> to about <NUM>. The stirring time at about <NUM> to about <NUM> is generally about <NUM> minutes to about <NUM> hours, and preferably about <NUM> hours to about <NUM> hours. The cooling temperature is room temperature. The stirring time under the cooling condition is generally from about <NUM> minutes to about <NUM> hours, and preferably from about <NUM> minutes to about <NUM> hours. The crystals from the suspension can be separated by methods known per se such as filtration or the like. The filtration temperature is room temperature, and preferably about <NUM> to about <NUM>.

Alternatively, A method in which the suspension is stirred at about <NUM> to about <NUM>, and then filtered at about <NUM> to about <NUM> to collect the crystal may be adopted.

The obtained crystal may be dried by a method known per se to obtain the crystal of the present invention. The drying may be carried out by reducing the pressure or by ventilation. The drying temperature is preferably no higher than about <NUM>, and more preferably about <NUM> to about <NUM>.

The crystal other than the crystal disclosed herein can be prepared by other known methods.

In order to analyze the obtained crystal, an X-ray diffraction crystallization analysis method is generally used. Furthermore, the crystal orientation can also be determined by a mechanical or optical method (e.g., FT-Raman spectroscopy, solid-state NMR spectroscopy, etc.).

The peaks of the spectra obtained through the afore-mentioned analysis method inevitably produce a certain determination error in nature. A crystal having a spectral peak within the error range is also encompassed in the concept of crystal disclosed herein.

For example, "± <NUM>" in the description of the interplanar crystal spacing (d) of the powder X-ray diffraction means that the error can be allowed.

The compound NB001 was synthesized according to the following synthetic route.

The compound <NUM> (<NUM>, <NUM> mmol, <NUM> eq) was dissolved in dioxane (<NUM>), and then SOCl<NUM> (<NUM>, <NUM> mmol, <NUM>, <NUM> eq) was slowly added to the afore-mentioned reaction solution, and the mixture was continuously stirred at <NUM> for <NUM> hours. The LCMS assay showed the raw materials are reacted completely and the desired product was generated. The solvent in the reaction solution was removed under reduced pressure in a water pump. The gray residue was added into <NUM> of ethanol and stirred for <NUM> minutes. The mixture was filtered through a sintered glass funnel. <NUM> of saturated sodium carbonate solution was added to the filtered solid and stirred for <NUM> minutes. The mixture was filtered through a sintered glass funnel. The filtered solid was spin-dried in a water pump under reduced pressure to obtain crude intermediate <NUM> (<NUM>, <NUM> mmol, with a yield of <NUM>% and a purity of <NUM>%), and the product is directly used in the next step without further purification.

The compound <NUM> (<NUM>, <NUM> mmol, <NUM> eq) was dissolved in n-BuOH (<NUM>), and then the compound <NUM> (<NUM>, <NUM> mmol, <NUM> eq) was slowly added to the afore-mentioned reaction solution, and the mixture was continuously stirred at <NUM> for <NUM> hours. The LCMS assay showed the raw materials are reacted completely and the desired product was generated. The solvent was removed under reduced pressure through a water pump, and the residue was concentrated to obtain a yellow crude product. <NUM> of DMF was added into the yellow crude product, then the mixture was continuously stirred at -<NUM> for <NUM> hour. Then the mixture was filtered through a sintered glass funnel. <NUM> of ethyl acetate was added into the filtered solid, and the mixture was filtered again to obtain an off-white solid NB001 (<NUM>, <NUM> mmol, with a yield of <NUM>% and a purity <NUM>%).

Into <NUM> vials were weighed <NUM> of the raw material respectively. Then appropriate amount of methanol, ethanol, isopropyl alcohol, n-butanol, acetonitrile, acetone, ethyl acetate, <NUM>-methyl tetrahydrofuran, tetrahydrofuran, dioxane, n-pentyl alcohol, toluene, isopropyl alcohol + water (with a mass ratio of <NUM>:<NUM>), acetonitrile + water (with a mass ratio of <NUM>:<NUM>), acetone + water (with a mass ratio of <NUM>: <NUM>), and ethanol + water (with a mass ratio of <NUM>:<NUM>) were added into respective vials, and the mixture was shaken rapidly to dissolve the compound of the mixture until the solution becomes clear. The approximate solubility of the resultant bulk pharmaceutical chemicals (BPC) in an organic solvent and a mixed solvent was shown in Table <NUM> below.

Into <NUM> vials were weighed <NUM> BPCs according to Table <NUM> respectively. Then appropriate amount of acetonitrile, tetrahydrofuran, acetone, isopropyl alcohol, ethyl acetate, and ethanol were added into respective vials to obtain a suspension. The suspension was stirred in a mixer under a condition of <NUM> for two days, and then dried by baking at <NUM> after centrifugation. Methanol, ethanol + water (with the mass ratio of <NUM>:<NUM>), isopropanol + water (with the mass ratio of <NUM>:<NUM>), and acetonitrile + water (with the mass ratio of <NUM>:<NUM>) were naturally evaporated <NUM> days after complete dissolution. The resultant solids were subjected to XRPD respectively and then compared with the XRPD of BPCs.

The X-ray powder diffractometer (XRPD) as used is the PANalytical Empyrean, with specific parameters of: tube: copper: K-Alpha (λ = <NUM>Ǻ); generator: voltage: 40kV, current: 40mA; scanning range: <NUM>-<NUM> degrees; sample rotation speed: <NUM> rpm; scanning speed: <NUM> degrees/min, and the results were as shown in <FIG>.

The physical characterization of the BPCs was determined using PLM, TGA, DSC, and DVS. The results are shown in <FIG>.

The solid samples were dispersed in silicone oil and observed under a PLM using a 10x ocular lens and a <NUM>/50x object lens. The microscope as used is Polarized Light Microscope-Nikon Eclipse LV 100POL from Nikon, and the images of the samples under the 20x object lens of this microscope were as shown in <FIG>.

<NUM> to <NUM> of the samples were placed in sample trays and heated from room temperature to <NUM> at a rate of <NUM>/min. The results were as shown in <FIG>.

DSC Assay: appropriate amount of samples was placed in specific aluminum trays and heated from <NUM> to <NUM> at <NUM> /min. The results were as shown in <FIG>.

Onto the sample trays were weighed about <NUM> of samples, and then the sample trays are put into the instrument for testing.

Stress test, long-term stable test and accelerated stability test of the compound with the crystal form:
The stability of the compound with such a crystal form was investigated under conditions of a high temperature (<NUM>), high humidity (<NUM>% RH), strong light irradiation (<NUM> Klx), <NUM>/<NUM>% RH (for accelerated test) and <NUM>/<NUM>% RH (for long term test), in accordance with "<NPL>).

<NUM> of the crystalline sample involved in the Examples of the present invention was accurately weighed and placed on the bottom of a glass sample vial, and then spread out to form a thin layer. The samples for test under high temperature and high humidity conditions and for the long-term test were contained in vials sealed with aluminum-foil paper, and some small holes were made on the aluminum-foil paper to ensure that the sample can be fully in contact with ambient conditions; The samples for test under strong light irradiation and for the accelerated test were contained in vials sealed with screw bottle caps. The conditions and timing for sample placing were shown in Table <NUM>. The samples placed under different conditions were taken and analyzed on day <NUM>, <NUM>, and <NUM>, and the analysis results were compared with the initial test results on day <NUM>. The results of the stability test were shown in Table <NUM>.

The above results showed that the stability of the compound with such a crystal form under various test conditions is good.

PWL determination at <NUM> after the 7th administration (21d): as compared with the model group, the non-crystalline NB001 group (<NUM>/kg, i. ); the group of the crystal dislcosed herein (<NUM>/kg, i. ); morphine (<NUM>/kg, i. ); and gabapentin (<NUM>/kg, i. ) all had a better analgesic effect, wherein it can be seen from <FIG> that the analgesic effect of the group of the crystal disclosed herein (<NUM>/kg, i. ) was extremely significant (P < <NUM>).

The visceral pain is induced by injecting yeast polysaccharide into the mice colony, and the yeast polysaccharide is derived from Saccharomyces cerevisiae (Sigma), wherein one kind of dextran is resident on the cell wall of the yeast and is identified as a protein-carbohydrate complex. Specifically, the mice were anesthetized after inhaling <NUM>-<NUM>% isoflurane, and then a <NUM> suspension of yeast polysaccharide (formulated as a <NUM>/ml solution in physiological saline) was injected into the colon of the mice by using a <NUM> long plastic feed needle within <NUM> minutes. The control group was injected with <NUM> physiological saline. The yeast polysaccharide or physiological saline was injected daily for <NUM> days. A visceral pain behavior test was carried out according to the Laird procedure. The number of times the mice licking the abdomen within <NUM> minutes was recorded, excluding other grooming behaviors, stretching of whole body, pressing the abdomen against the floor, and an <NUM>-<NUM>-second arched posture. On day <NUM>, <NUM> and <NUM> after the injection into the colon, an open field test was carried out from <NUM> am to <NUM> pm, and the entire test was a double-blind test. Prior to the behavioral test, the mice were acclimated for <NUM> minutes in an observation room. The mice were placed at the middle of a new open zone (<NUM> × <NUM> × <NUM><NUM>) with micro light (<<NUM> lux) and a fan. The movement distance, vertical count, movement count, stereotypic behavior count, and jumping count of animals were recorded within <NUM> minutes by means of an activity monitoring system equipped with multiple pairs of packaged light beam sets.

An anxious animal model of the prior art - the elevated plus maze test (EPM) was adopted. The mice were acclimated for <NUM> minutes in the room prior to the behavior observation. The EPM was composed of two open arms (<NUM> lux) and two closed arms (<NUM> lux) arranged opposite to each other. For each test, an individual animal was placed at the center of the test position and allowed to move freely. The test is carried out for <NUM> minutes. The number of entering the test position for each arm and the time spent therefore were recorded.

Bright/dark box test: The test procedure was carried out by using an improved method in accordance with the prior art. The device was composed of a rectangular organic glass box (<NUM> × <NUM> × <NUM><NUM>), which is separated into bright and dark compartments with the equal size by using a door. A <NUM> watt lamp (<NUM> lux) in turn-on state was placed at <NUM> above the box to irradiate the bright compartment. Each animal was first placed in the dark compartment for <NUM> seconds, and then the door leading to the bright compartment was open. The time each animal stayed in the dark compartment and the time each animal enters the bright compartment within <NUM> minutes were recorded.

The physiological saline, non-crystalline NB001 (<NUM>/kg), the crystal disclosed herein (<NUM>/kg), and Gabapentin (<NUM>/kg) as a positive control were intraperitoneally injected (IP) into the mice treated with yeast polysaccharide respectively. The behaviors of Mice were observed <NUM> days after the yeast polysaccharide treatment. The results were shown in Table <NUM>, indicating that as compared with the positive control group, the non-crystalline NB001 (<NUM>/kg), the crystal disclosed herein (<NUM>/kg) both had significant therapeutic effects. Furthermore, the therapeutic effect of the group of the crystal disclosed herein is extremely significant.

The other two sets of tests were conducted to test the effect of the crystal disclosed herein on inhibiting anxiety caused by chronic pain and neuropathic pain.

The EPM test tested the anxiety behavior of mice after the test treatment. The physiological saline, non-crystalline NB001 (<NUM>/kg), the crystal disclosed herein (<NUM>/kg), and Gabapentin (<NUM>/kg) as a positive control were intraperitoneally injected (IP) into the mice treated with yeast polysaccharide respectively. The behaviors of Mice were observed <NUM> days after the yeast polysaccharide treatment. The results were shown in Table <NUM>, indicating that as compared with the positive control group, the non-crystalline NB001 (<NUM>/kg), the crystal disclosed herein (<NUM>/kg) both had significant therapeutic effects. Furthermore, the therapeutic effect of the group of the crystal disclosed herein is extremely significant.

In order to confirm the effect of the crystal disclosed herein on inhibiting anxiety, mice were subjected to a bright/dark box test, and the results were as shown in Table <NUM>. The physiological saline, non-crystalline NB001 (<NUM>/kg), the crystal disclosed herein (<NUM>/kg), and Gabapentin (<NUM>/kg) as a positive control were intraperitoneally injected (IP) into the mice treated with yeast polysaccharide respectively. The behaviors of Mice were observed <NUM> days after the yeast polysaccharide treatment. The results were shown in Table <NUM>, indicating that as compared with the positive control group, the non-crystalline NB001 (<NUM>/kg), the crystal dislcosed herein (<NUM>/kg) both had significant therapeutic effects. Furthermore, the therapeutic effect of the group of the crystal disclosed herein is extremely significant.

Claim 1:
Method for the preparation of a crystal of <NUM>-((<NUM>-(<NUM>-amino-<NUM>-purin-<NUM>-yl)ethyl)amino)pentan-<NUM>-ol,
wherein the crystal is measured by using Cu-Ka and shows a characteristic peak at an interplanar crystal spacing d of <NUM>°±<NUM>°, <NUM>°±<NUM>°, <NUM>°±<NUM>°, <NUM>°±<NUM>°, <NUM>°±<NUM>°, <NUM>°±<NUM>°, <NUM>°±<NUM>°, <NUM>°±<NUM>° and <NUM>°±<NUM>° in powder x ray diffraction,
characterized in that said method comprising the synthesis route as below:
<CHM>
and a subsequent crystal transition method selected from the group consisting of crystallization of solution, vapor crystallization, a melt crystallization method, a special growth method, a stream fog method and a slurry method.