WO2017067438A1 - Crystal form of anamorelin and preparation method therefor - Google Patents

Abstract

The present invention provides a crystal form of anamorelin and a preparation method therefor.

Description

Crystal form of alamorelin and preparation method thereof Technical field

The present invention relates to a kind of (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide New crystal form and its preparation method.

Background technique

Anamorelin, whose chemical name is: (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2- Trimethylformylhydrazide is a compound that increases mammalian growth hormone levels and has a compound structure as shown in Formula I:

Cancer cachexia is a state of consumption in which patients lose a lot of weight and muscle mass. It is necessary for the treatment of cachexia because it weakens the patient, affects the quality of life and interferes with the patient's treatment plan. The drug alamorelin produces the same effect as the so-called "starved hormone" ghrelin, which stimulates hunger. Alamolin is a mimetic of ghrelin, which is secreted by the stomach and is a ligand for growth hormone receptors. . Alamolin binds to this receptor, causing the release of growth hormone, causing a metabolic cascade that affects a variety of different factors, including fat-removing body weight, as well as blood sugar metabolism. Therefore, alamorelin can also enhance the appetite of patients and help patients stay healthy. The 2014 European Society of Medical Oncology (ESMO) in Madrid, Spain, announced that Alamolin is expected to be the first drug in history to effectively improve cancer cachexia.

Alamolin is a drug developed by Helsinn Therapeutics (Switzerland) from Novo Nordisk for the development of a cachexia and anorexia for patients with cancer, including non-small cell lung cancer. It can also be used to treat hip fractures and preventive diseases. The strength of the elderly and the elderly has continued to decline. In two key, 12-week Phase III clinical trials (ROMANA 1, ROMANA 2), alamorelin can significantly increase the body fat loss, and is generally tolerated; the incidence of serious adverse drug reactions is less than 3%, mainly related to hyperglycemia and diabetes. Compared with the placebo group, alamorelin continued to increase body weight and improve cancer anorexia-cachexia-related symptoms and concerns; however, there was no significant difference in the improvement of grip strength between the alamolin group and the placebo group. Therefore, this product has excellent clinical value and market value.

The polymorphic form of the drug free base and its preparation are reported as follows:

Synthesis of (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylmethyl is disclosed in the patent ZL99806010.0 A method for synthesizing hydrazide, and using [(1R)-2-[(3R)-3-benzyl-3-(N,N',N'-trimethylmethylcarbonyl)piperidin-1-yl tert-Butyl ester of 1-((1H-indol-3-yl)methyl)-2-oxoethyl]carbamate is dissolved in dichloromethane, then trifluoroacetic acid is added to remove tert-butyl formate After the base, the mixture was concentrated to remove the solvent, and then the product was extracted with dichloromethane, and the obtained extract was concentrated to dryness to give (3R)-1-(2-methylalanyl-D-color ammonia as an amorphous powder. Acyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide.

Patent ZL00815145.8 discloses the synthesis of alamorelin and its compounds as pharmaceutically acceptable salts, relating to novel diastereomeric compounds, pharmaceutically acceptable salts thereof, compositions containing them and their use in therapy Lack of use of medical conditions caused by growth hormone. Synthesis of (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformyl is disclosed in this patent. The synthesis method of hydrazine, and using [(1R)-2-[(3R)-3-benzyl-3-(N,N',N'-trimethylmethylcarbonylcarbonyl)piperidin-1-yl] 1-((1H-Indol-3-yl)methyl)-2-oxoethyl]carbamic acid tert-butyl ester was dissolved in ethyl acetate, and then hydrogen chloride gas was passed to remove the tert-butyl formate protection group. , the solid is dissolved in water, and then the pH is adjusted to about 7 with sodium carbonate, and the product is extracted with dichloromethane; the extract phase is concentrated to obtain (3R)-1-(2-methylalanyl-D-tryptophan). -3-Benzyl-3-piperidine 1,2,2-trimethylformylhydrazide.

Patent WO2006016995 discloses (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylmethyl as a medicament Crystalline polymorphs of hydrazides, methods of producing and separating these polymorphs, and pharmaceutical compositions and drug therapies containing these polymorphs, the crystalline polymorphs for direct application to the pituitary Gland cells release the growth hormone. This patent discloses (4R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazone 4 Crystal form: Form A, Form B, Form C and Form D. The patent also provides the preparation of 3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazone. The method of crystal form, especially the preparation method of Form C, in which the method of removing the tert-butyl formate protecting group of methanesulfonic acid in methanol is utilized without exception. As a well-known cause in the art, clinical studies have found that mesylate is genotoxic, and its DNA alkylation leads to mutagenic effects, in which methyl methanesulfonate and ethyl methanesulfonate have been reported. (eg document EMEA/44714/2008). The invention adopts hydrochloric acid or hydrogen chloride gas to remove the tert-butyl formate protecting group, avoids the method of removing methanesulfonic acid, thereby avoiding the risk of the genotoxic impurities in the process, and increasing the risk. The safety of the drug.

(3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-three prepared by the patent ZL99806010.0 and the patent ZL00815145.8 Methyl formyl hydrazide, no data on the purity of its compounds, we found that (3R)-1-(2-methylalanyl-D-tryptophan)-3 was prepared by this method. -Benzyl-3-piperidine 1,2,2-trimethylformylhydrazide does not help to remove the impurities produced, and the purity of the obtained product is not high, and it is difficult to meet the medicinal requirements. And (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethyl obtained by the preparation method of the present invention. The crystal form of the formyl hydrazide has a purity of 99.8% and a single impurity of less than 0.1%, which fully meets the requirements for medicinal purity. Moreover, the crystal form is stable to conditions such as pressure, temperature, humidity and illumination, and the preparation method is simple in operation and suitable for industrial production.

Summary of the invention

The present invention provides a (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-tricarboxylate of formula I. The crystal form of methyl formyl hydrazide characterized by X-ray powder diffraction pattern expressed by 2θ angle and interplanar spacing using Cu-Kα1 radiation, the crystal having a characteristic absorption peak at 2θ±0.20 as follows : 6.50, 9.20, 10.68, 12.74, 14.58, 15.30, 16.99, 18.49, 19.63, 20.70,

The crystalline form may be in the form of a hydrate or solvated form, all of which are included within the scope of the invention. Preferably the crystalline form is a hydrate, preferably a monohydrate.

Preferably, the Form E of the present invention has an X-ray powder diffraction pattern as shown in FIG. The location of the characteristic peaks and their intensities in the figure are listed in the following table:

Peak number 2θ value D Relative Strength NO. (degree) (A) % 1 6.50 13.58 100 2 9.20 9.60 82.6 3 10.68 8.27 22.0 4 12.74 6.94 29.4 5 13.83 6.39 17.0 6 14.58 6.07 44.9 7 15.30 5.78 32.6 8 15.86 5.58 3.8 9 16.99 5.21 25.2 10 18.49 4.79 21.0 11 19.63 4.51 24.6 12 20.70 4.28 29.0 13 21.63 4.10 15.2 14 22.47 3.95 17.2 15 23.67 3.75 10.2 16 24.49 3.63 5.8 17 25.62 3.47 1.6 18 27.08 3.29 7.4 19 29.30 3.04 8.4

20 32.43 2.75 7.0 twenty one 37.06 2.42 1.3 twenty two 38.69 2.32 1.3 twenty three 39.69 2.26 1.2

Another object of the present invention is to provide a suitable industrial preparation of (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2- A method of crystal form of trimethylformylhydrazine E. In particular, the present invention provides two different methods for efficiently obtaining the E crystal form.

The first method provided by the present invention comprises (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethyl a step of dissolving a hydrazide or a solvate thereof in a mixed solvent of an organic solvent and water, cooling, and crystallization, preferably, the organic solvent is selected from an alcohol having less than 5 carbon atoms or other polar solvent, more preferably Selected from methanol, ethanol, isopropanol, butanol, ethylene glycol, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, acetone , butanone, methyl ethyl ketone, methyl isobutyl ketone, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane.

The mixed solvent may be added separately, specifically, for example, according to (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2 , 2-trimethylformylhydrazide is first dissolved in an organic solvent and then added with water, or (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3 - Piperidine 1,2,2-trimethylformylhydrazine is first suspended in water and then added as an organic solvent. Preferably, (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide is dissolved in the mixture The solvent is then warmed to a temperature of 30 to 90 bases, preferably 40 to 80 ° C, most preferably 50 to 70 ° C, and stirred at this temperature for a period of time.

In the method, the (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylmethyl The weight ratio of the hydrazide to the mixed solvent may be from 1:1 to 20.

The ratio of water to organic solvent is from 5% to 95% V/V, preferably from 20% to 80% V/V, more preferably from 30% to 70% V/V, most preferably from 40% to 60% V/V.

Another preparation method provided by the present invention comprises (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2- The acid addition salt of trimethylformylhydrazine is dissolved in water, and then a base is added to carry out a step of crystallization after neutralization.

The acid addition salt is an inorganic or organic acid, preferably selected from the group consisting of hydrochloride, hydrobromide, sulfate, acetate, phosphate, fumarate, maleate, mandelate, O-phenylene Formate, glutarate, methanesulfonate, salicylate, succinate, tartrate, besylate, tosylate.

The base is an inorganic base or an organic base, preferably selected from the group consisting of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydrogencarbonate, dipotassium hydrogen sulfate, aqueous ammonia, triethylamine, N-methyl. Morpholine, diisopropylethylamine, diethylamine.

In the method, preferably, the step of adjusting the pH is further included, and the pH ranges from 6 to 14.

In the above two steps, the crystallization may be carried out under static conditions or under stirring; the crystallization method is a conventional method in the art, such as cooling, distilling off part of the solvent, adding anti-solvent, and adding crystal Single or combined use of methods. Wherein "anti-solvent" means (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-three at normal temperature. The solubility of methylformylhydrazide is not good, but it can dissolve 3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine with step 1). A solvent-miscible solvent of 1,2,2-trimethylformylhydrazide, such as water, n-hexane, n-heptane, and the like.

In the above two steps, it is preferred to further comprise the steps of separating and drying.

Separation may be carried out by conventional methods in the art, such as filtration, centrifugation, etc. Alternatively, the separated solid may be washed with the corresponding solvent used in the above steps.

The drying temperature is usually from 30 ° C to 100 ° C, preferably from 40 ° C to 70 ° C, and it can be dried at normal pressure or dried under reduced pressure. Preferably, the drying is dried under reduced pressure.

Among the above two methods, seed crystals can be added.

In a further aspect of the invention there is provided a pharmaceutical composition comprising the aforementioned crystalline form E and a pharmaceutically acceptable carrier.

The crystalline form E of the present invention or the aforementioned pharmaceutical composition can be used for the preparation of a medicament for directly acting on pituitary gland cells to release growth hormone.

It has been experimentally proved that the E crystal form of the present invention has higher stability with higher stability than the amorphous state and other crystal forms of the compound, such as HPLC area normalization purity of 98%, 99% or More than 99.5%, due to the high stability and high purity of these crystal forms, they can achieve the more stringent drug regulation and specifications normally required for pharmaceutical preparations: these advantages are beneficial to them to make corresponding Formulations, such as their formulations, have good stability and effectiveness in preparation and storage; on the other hand, they are also useful for making high purity acid adducts, such as they can be made from. The HPLC area normalization method has a purity of 98%, 99% or more, and a single impurity is less than 0.15%, 0.10% or 0.05% of the anamorelin hydrochloride and the fumarate acid addition product. Used for preparation of preparations and the like.

Compared to the prior art, the invention has unique advantages:

1) The (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethyl group obtained by the present invention Formyl hydrazide E crystal form, with stable physicochemical properties, stable conditions such as pressure, temperature, humidity and light; from the DSC analysis, the crystal form of the publicly reported crystal form shows that the DSC pattern is 100 ° C and then 116.8 ° C The melting endotherm of the peak, while the E-type crystal of the present invention has a DSC showing a melting endotherm which starts at 120.05 ° C and then peaks at 127.91 ° C, and there is also a significant difference. From this aspect, the new crystal form of the present invention has more High melting point value, within a certain range, this product has higher thermal stability.

2) The (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethyl group obtained by the present invention Formyl hydrazide E crystal form, has a simple and feasible preparation method: after adopting the preparation process of the invention, the operation is simple, easy to obtain and separate, and the stable physical and chemical properties can be dried at normal pressure or under reduced pressure, and Therefore, large-scale production can be carried out. At the same time, its good stability makes this product do not require special storage conditions, these crystal forms can be stored for a long time and stable.

3) The (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethyl group prepared by the present invention. Formyl hydrazine E crystal form, with high purity, its purity can reach 99.8%, single impurity is less than 0.1%, fully meet the requirements of medicinal purity.

4) The (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethyl group prepared by the present invention. Formyl hydrazide E crystal form avoids the generation of genotoxic impurities, reduces the risk of toxic impurities on the drug, and increases the safety of the drug.

DRAWINGS

The accompanying drawings, which are incorporated in the claims

Figure 1 (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine E crystal powder X-ray diffraction spectrum;

Figure 2 (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazone E crystal Differential thermal analysis DSC spectrum;

Figure 3 (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazone E crystal Thermogravimetric analysis of TGA spectra.

detailed description

The above description of the present invention will be further described in detail with reference to the specific embodiments of the embodiments of the present invention. The scope of the topic is limited to the following examples. Various alterations and modifications may be made without departing from the spirit and scope of the invention.

Test instrument used in the experiment

1. X-ray diffraction spectrum

Instrument model: Bruker D8FOCUS X-ray powder diffractometer (Brook, Germany)

Ray: Monochrome Cu-Kα1 ray

Scanning mode: θ/2θ, scanning range: 2 to 40°

Voltage: 40KV, current: 40mA

2, DSC spectrum

Instrument model: METTLER TOLEDO DSC1 (METTLER TOLEDO)

Purge gas: nitrogen

Heating rate: 10.00K/min

Temperature range: 40～300°C

3, TGA

Instrument model: Mettler Toledo DSC1/TG209F3 (METTLER TOLEDO)

Purge gas: nitrogen

Heating rate: 20.00K/min

Temperature range: 40-700 ° C

Synthesis of Amorphous (3R)-1-(2-Methylalanyl-D-tryptophanyl)-3-benzyl-3-piperidine 1,2,2-Trimethylformylhydrazide

Example 1:

300 g of [(1R)-2-[(3R)-3-benzyl-3-(N,N',N'-trimethylcarbamidocarbonyl)piperidin-1-yl]-1-(( 1H-Indol-3-yl)methyl)-2-oxoethyl]carbamic acid tert-butyl ester was added to the reaction flask, and then 4 L of dichloromethane was added to the reaction flask, and the raw material was completely dissolved by stirring.

Then, the reaction system is cooled to 10 ° C or lower in an ice bath, hydrogen chloride gas is continuously supplied to the reaction liquid, and solids are gradually precipitated, and the reaction is further maintained at about 10 ° C for 3 to 5 hours, and the sample is detected. After the reaction of the raw materials is completed, the reaction system is completed. 1.5 L of water was added thereto, the solid was completely dissolved, and then the pH was adjusted to about 8 with a 20% aqueous sodium hydroxide solution, and the layers were separated; the aqueous phase was extracted once more with dichloromethane, and the organic phases were combined.

The organic phase was dried over anhydrous sodium sulfate for 3 hrs, filtered, and then evaporated to ethylamine 3-Benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude 246 g, yield 97.2%. HPLC content (area normalization method) was 96.1%.

Example 2:

300 g of [(1R)-2-[(3R)-3-benzyl-3-(N,N',N'-trimethylcarbamidocarbonyl)piperidin-1-yl]-1-(( 1H-Indol-3-yl)methyl)-2-oxoethyl]carbamic acid tert-butyl ester was added to the reaction flask, 36% concentrated hydrochloric acid was added to the reaction flask, and the reaction system was heated to 40 with stirring. The reaction was carried out at ° C to 50 for 3 hours.

Then, the sample is detected. After the reaction of the raw material is completed, the reaction system is cooled to 10 or less, and 2.0 L of dichloromethane is added to the reaction system, and then the pH is adjusted to about 8 with a 20% aqueous sodium hydroxide solution, and the aqueous phase is further separated. It was extracted once with dichloromethane and the organic phases were combined.

The organic phase was dried over anhydrous sodium sulfate for 3 hrs, filtered, and then evaporated to ethylamine 3-Benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude 248 g, yield 98%. HPLC content (area normalization method) was 96.2%.

Preparation of (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazone E crystal form

Example 3

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10g was added to the reaction flask and 30 ml of N- was added.

Methylpyrrolidone, stirred and dissolved completely. Then, 60 ml of water was added dropwise to the reaction flask at room temperature, and the reaction liquid was heated to 60 ° C. The solution became cloudy, and a white solid was gradually precipitated, and stirring was continued for 2 hours.

Slowly cooled to below 20 ° C, filtered, and the filter cake was washed with a mixture of N-methylpyrrolidone / H ₂ O; the cake was vacuum dried at about 55 ° C to obtain (3R)-1-(2-methylalanyl) -D-tryptophan)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 9.5 g), HPLC content (area normalization) 99.72%. The XRD pattern is shown in Fig. 1, the DSC chart is shown in Fig. 2, and the TGA pattern is shown in Fig. 3, where the crystal form is defined as the E crystal form. The DSC of the crystal form has an endotherm at 120.05, the TGA is heated at 60A, and the crystal loss of 5 is about 3.1%. Combined with the Karl Fischer method, the moisture content of the product is determined. 3.1% and 3.2% indicate that the sample is present as a monohydrate.

Example 4:

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 30 ml of N,N-dimethylformamide was added, stirred, and dissolved completely. Then, 30 ml of water was added dropwise to the reaction flask at room temperature, and the reaction solution was heated to 50 ° C. The solution became cloudy, and a white solid was gradually precipitated, and stirring was continued for 2 h.

Slowly cool to below 10 ° C, filter, filter cake washed with N, N-dimethylformamide / H ₂ O mixture; vacuum cake dried at around 55 ° C to obtain (3R)-1-(2-A Alanyl-D-tryptophanyl-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 8.5 g), HPLC content (area normalization) ) 99.87%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 5:

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 30 ml of dimethyl sulfoxide was added, stirred, and dissolved completely. Then, 40 ml of water was added dropwise to the reaction flask at room temperature, and the reaction liquid was heated to 60 ° C, the solution became cloudy, and a white solid was gradually precipitated, and stirring was continued for 2 hours.

Slowly cooled to below 10 ° C, filtered, and the filter cake was washed with a mixture of dimethyl sulfoxide / H ₂ O; the cake was vacuum dried at about 50 ° C to obtain (3R)-1-(2-methylalanyl) -D-tryptophanyl-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 9.1 g), HPLC content (area normalization) 99.61%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 6

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 40 ml of 1,4-dioxane was added, stirred, and dissolved completely. Then, 50 ml of water was added dropwise to the reaction flask at room temperature, and the reaction solution was heated to 70 ° C. The solution became cloudy, and a white solid was gradually precipitated, and stirring was continued for 2 hours.

Slowly cooled to below 10 ° C, filtered, and the filter cake was washed with a mixture of 1,4-dioxane/H ₂ O; the cake was vacuum dried at about 50 ° C to obtain (3R)-1-(2-methyl alanyl-D-tryptophan-3-

Benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 8.7 g), HPLC content (area normalization) 99.11%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 7

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 40 ml of N,N-dimethylacetamide was added, stirred, and dissolved completely. Then, 40 ml of water was added dropwise to the reaction flask at room temperature, and the reaction solution was heated to 70 ° C. The solution became cloudy, and was slowly cooled to about 50 ° C. Seed crystals were added thereto, and cooling was continued to gradually precipitate a solid.

The reaction system was cooled to about 10 ° C, filtered, and the filter cake was washed with a mixture of N,N-dimethylacetamide/H ₂ O; the cake was vacuum dried at about 50 ° C to obtain (3R)-1-(2- Methylalanyl-D-tryptophanyl-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 8.1 g), HPLC content (area normalized) Law) 99.78%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 7

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 50 ml of acetone was added, stirred, and dissolved completely. Then, 70 ml of water was added dropwise to the reaction flask at room temperature, and the reaction liquid was heated to 45 ° C. The solution became cloudy, and a white solid was gradually precipitated, and stirring was continued for 2 hours.

Slowly cool to below 10 ° C, filter, filter cake washed with acetone / H ₂ O mixture; filter cake vacuum dried at around 50 ° C to obtain (3R)-1-(2-methylalanyl-D-color Aminoacyl-3-phenylmethyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 9.3 g), HPLC content (area normalization) 98.9%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 9

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 50 ml of ethanol was added, stirred, and dissolved completely. Then, 60 ml of water was added dropwise to the reaction flask at room temperature, and the reaction solution was heated to 60 ° C. The solution became cloudy. After stirring for 2 hours, the mixture was cooled to 40 ° C, seed crystals were added, and cooling was continued to precipitate a solid.

Cool to about 0 ° C, filter, filter cake washed with ethanol / H ₂ O mixture; filter cake dried at around 50 ° C to obtain (3R)-1-(2-methylalanyl-D-tryptoyl --3-Benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 9.5 g), HPLC content (area normalization) 99.05%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 10:

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 30 ml of tetrahydrofuran was added, stirred, and dissolved completely. Then, 45 ml of water was added dropwise to the reaction flask at room temperature, and the reaction liquid was heated to 60 ° C, and the solution became cloudy. After stirring for 2 hours, a solid was precipitated.

Cool to about 0 ° C, filter, filter cake washed with ethanol / H ₂ O mixture; filter cake dried at around 50 ° C to obtain (3R)-1-(2-methylalanyl-D-tryptoyl --3-Benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 8.8 g), HPLC content (area normalization) 98.95%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 11

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10 g was added to the reaction flask, 30 ml of water was added, the crude product was suspended in water, 30 ml of N,N-dimethylformamide was added to the reaction, and the reaction mixture was heated to 55 ° C. The solution became cloudy and gradually precipitated a white solid. Continue to stir at constant temperature for 2 h.

Slowly cooled to below 10 ° C, filtered, and the filter cake was washed with a mixture of N,N-dimethylformamide/H ₂ O; the cake was vacuum dried at about 55 ° C to obtain (3R)-1-(2- Methylalanyl-D-tryptophanyl-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 8.6 g), HPLC content (area normalized) Law) 99.70%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 12

Taking the above amorphous (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine crude product 10g was added to the reaction flask and 200 ml of acetic acid was added.

The ethyl ester was stirred and dissolved, and the temperature was controlled at about 20 ° C. Fumaric acid (2.0 g) was suspended in 50 ml of ethyl acetate, and then slowly added dropwise to the reaction system with stirring to precipitate a solid.

After the dropwise addition was completed, stirring was continued for about 1 hour at 20 ° C, and then filtered. The filter cake was washed with ethyl acetate and dried to give (3R)-1-(2-methylalanyl-D-tryptoyl)-3-benzyl-3-piperidine 1,2,2 Trimethylformyl hydrazide fumarate (white solid, 10.3 g).

(3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformyl hydrazide fumarate 10.3 g was added to the reaction flask, then 40 ml of water was added, stirred, and the solid was completely dissolved. A solution of sodium hydroxide (2.17 g) in water (20 ml) was added dropwise to the reaction mixture, and the pH was adjusted to 7 to 8 to precipitate a white solid. After stirring for 1 hour, it was filtered. Filter cake

After washing with water, it is dried (55 ° C) to obtain (3R)-1-(2-methylalanyl-D-tryptophyl)-3- Benzyl-3-piperidine 1,2,2-trimethylformylhydrazine (white solid, 9.1 g), HPLC content (area normalization) 97.05%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 13

The (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-triate containing the E crystal form obtained in the above Example 10 was obtained. 4 g of crude methylformylhydrazine was added to the reaction flask, 12 ml of N,N-dimethylformamide was added, stirred, and dissolved completely. Then, 15 ml of water was added dropwise to the reaction flask at room temperature, and the reaction liquid was heated to 50 ° C. The solution became cloudy, and a white solid was gradually precipitated, and stirring was continued for 2 hours.

Slowly cooled to below 10 ° C, filtered, and the filter cake was washed with a mixture of N,N-dimethylformamide/H ₂ O; the cake was vacuum dried at about 50 ° C to obtain (3R)-1-(2-A Alanyl-D-tryptophanyl-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide (white solid, 3.4 g), HPLC content (area normalization) ) 99.75%. Upon comparison, it was confirmed that the solid was in the E crystal form.

Example 14

(3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylate prepared in the above examples. The 肼E crystal form crystal was dried under reduced pressure at 100 ° C for 24 hours, and the sample was subjected to X-RD ray powder diffraction and DSC analysis to find that the crystal form of the sample did not change, indicating that the crystal form has thermal stability.

Example 15

(3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylate prepared in the above examples. The crystal form of 肼E crystal was pressed at a pressure of 25 kg/cm ² , and the crystal form of the sample was not changed by X-RD ray powder diffraction and DSC analysis, indicating that the crystal form was stable under high pressure.

Example 16:

(3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylate prepared in the above examples.肼E crystal form crystals were tested at high temperature 40±2°C (0-60 days), light (0-60 days), high humidity 75%±5% (0-60 days), respectively. Related substances and crystal form detection.

The relevant substances are detected by HPLC, and the detection conditions are as follows:

RP analysis was performed on a 218 TP54 4.6 mm x 250 mm C18 silica column eluting at a flow rate of 1 ml/min at 42 ° C using UV detection at 214, 254, 276 and 301 nm. The column was equilibrated with 5% acetonitrile, 85% water and 10% aqueous 0.5% trifluoroacetic acid.

It was equilibrated from 5% (acetonitrile + 0.1% TFA) in aqueous TFA (0.1%). After the injection, the sample was eluted by a gradient solution of 5% to 60% (acetonitrile + 0.1% TFA) in the same aqueous buffer for 50 minutes. The content of the substance is calculated by the area normalization method.

Chiral HPLC analysis:

Chiral analysis was carried out using UV detection at 225 and 254 nm on a 4.6 mm x 250 mm Chirale 0J column packed with a 4.6 mm x 80 mm Chirale0J precolumn at room temperature at a flow rate of 0.7 ml/min. The sample was eluted with an isocratic eluate of heptane:isopropanol:trifluoroacetic acid (92:8:0.1).

The results of the survey are shown in the following table:

Claims (15)

(3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformyl group of formula I The E crystal form of ruthenium is characterized in that an X-ray powder diffraction pattern expressed by 2θ angle and interplanar spacing is obtained using Cu-Ka radiation, and the crystal has a characteristic absorption peak at 2θ±0.20 as follows: 6.50, 9.20 , 10.68, 12.74, 14.58, 15.30, 16.99, 18.49, 19.63, 20.70,

The crystal form of E according to claim 1, which has an X-ray powder diffraction pattern as shown in FIG. Crystal Form E according to claim 1, characterized in that the crystalline form is a hydrate, preferably a monohydrate. A process for the preparation of the E form according to any one of claims 1 to 3, which comprises (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzene a step of dissolving methyl-3-piperidine 1,2,2-trimethylformylhydrazine or a solvate thereof in a mixed solvent of an organic solvent and water, cooling, and crystallization, preferably, the organic solvent is selected from carbon An alcohol or other polar solvent having an atomic number of less than 5, more preferably selected from the group consisting of methanol, ethanol, isopropanol, butanol, ethylene glycol, N,N-dimethylformamide, N,N-dimethyl B Amide, N-methylpyrrolidone, dimethyl sulfoxide, acetone, methyl ethyl ketone, methyl ethyl ketone, methyl isobutyl ketone, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane. The production method according to claim 4, wherein said (3R)-1-(2- Methylalanyl-D-tryptophanyl-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazide is first dissolved in an organic solvent and then added to water, or (3R)- 1-(2-Methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2-trimethylformylhydrazine was first suspended in water and then added to an organic solvent. The method according to claim 4 or 5, wherein (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2- The trimethylformylhydrazide is heated to 30 to 90 ° C, preferably 40 to 80 ° C, and most preferably 50 to 70 ° C after being dissolved in the mixed solvent. The method according to claim 4, wherein said (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzyl-3-piperidine 1,2,2 The weight ratio of trimethylformylhydrazide to the mixed solvent is 1:1 to 20. The production method according to claim 4, wherein the ratio of the water to the organic solvent is 5% to 95% V/V. A process for the preparation of the E form according to any one of claims 1 to 3, which comprises (3R)-1-(2-methylalanyl-D-tryptophyl)-3-benzene The acid addition salt of methyl-3-piperidine 1,2,2-trimethylformylhydrazine is dissolved in water, and a step of neutralizing the crystal by adding a base is added. A process according to claim 9 wherein said acid addition salt is an inorganic or organic acid, preferably selected from the group consisting of hydrochloride, hydrobromide, sulfate, acetate, phosphate, fumarate. , maleate, mandelate, phthalate, glutarate, methanesulfonate, salicylate, succinate, tartrate, besylate, tosylate. The method according to claim 9, wherein said base is an inorganic base or an organic base, preferably selected from the group consisting of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydrogencarbonate, dipotassium hydrogen sulfate, Ammonia, triethylamine, N-methylmorpholine, diisopropylethylamine, diethylamine. The method of claim 9 further comprising the step of adjusting the pH, said pH being in the range of 6-14. The method of claim 4 or 9, further comprising the step of seeding. A pharmaceutical composition comprising the crystalline form E of any one of claims 1 to 3 and a pharmaceutically acceptable carrier. Use of the E crystal form according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 14 for the preparation of a medicament for directly acting on pituitary gland cells to release growth hormone, preferably for preparing a cancer patient Use in drugs for cachexia and anorexia.

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