WO2020186960A1 - Bulleyaconitine a crystalline form c, preparation method therefor and application thereof - Google Patents

Abstract

A bulleyaconitine A crystalline form C, and a preparation method therefor. An X-ray powder diffraction spectrum of the crystalline obtained by measurement using a Cu-Kα ray is as shown in fig. 1. The preparation method therefor comprises: using DMF as a solvent and obtaining by means of a gas-solid permeation method. The procedure of the preparation process is simple, and the obtained crystalline form is a solvate crystalline form, has high purity, and has good stability to light, humidity, and heat.

Description

A kind of aconitine C crystal form and its preparation method and application

This application claims the priority of the Chinese patent application filed with the Chinese Patent Office on March 15, 2019, the application number is 201910197723.8, and the invention title is "a crystal form of aconitine C and its preparation method and application", all of which The content is incorporated in this application by reference.

Technical field

The invention relates to the field of medicinal chemistry, in particular to a crystal form of aconitine C and its preparation method and application.

Background technique

The chemical name of oxaconitine is (1α, 6α, 14α, 16β) tetrahydro-8,13,14-triol-20-ethyl-1,6,16-trimethoxy-4-methoxymethyl- 8-Acetoxy-14-(4'-p-methoxybenzyl)-aconitine. It is a diterpene diester alkaloid extracted and isolated from the root tuber of Aconitum georgei Comber, a plant of the genus Aconitum in the Ranunculaceae family, named Crassicauline A. It was later renamed Bulleyaconitine A (T2), which is a known natural compound in plant species, and its structural formula is as follows:

At present, aconitine preparations are widely used clinically to treat rheumatoid arthritis (RA), osteoarthritis, myofibritis, neck and shoulder pain, low back pain, cancer pain and chronic pain caused by various reasons.

Drug polymorphism is a common phenomenon in drug development and an important factor affecting drug quality. The same drug with different crystal forms has differences in appearance, solubility, melting point, dissolution, and bioavailability, and may even have significant differences, which will affect the stability, bioavailability and efficacy of the drug. Moreover, the crystal form of the drug will also affect the quality of the pharmaceutical preparation of the drug, the absorption behavior in the human body, and ultimately affect the therapeutic effect of the preparation in the human body and the benefit ratio of side effects. With the in-depth research of aconitine A, the research on the crystal form and physicochemical properties of aconitine A is of great significance to the evaluation of the efficacy, quality and safety of aconitine A. The Chinese patent with application number 201710423005.9 discloses dissolving aconitine A with C1-C4 organic solvents, and the obtained aconitine solution is added dropwise to water, stirring while adding, after the addition, suction filtration, and the filter cake is dried. Obtained the amorphous grass Aconitum. At present, there is no relevant report on the crystalline aconitine.

Summary of the invention

In view of this, the purpose of the present invention is to provide a new crystal form of aconitine and its preparation method.

An object of the present invention is to research, discover and provide the crystal form C crystal form of aconitine by crystallographic methods.

The present invention uses the internationally recognized X-ray powder diffraction method (XRPD) to study and characterize the crystalline form of aconitine. Measurement conditions and methods: Cu/K-alpha1 (target), 45KV-40mA (working voltage and current), 2θ=3-40 (scanning range), scanning time per step (s) is 17.8-46.7, scanning step size ( 2θ) is 0.0167-0.0263,

The substantially pure crystal form C provided by the present invention has an X-ray powder diffraction pattern as shown in Figure 1, and its X-ray powder diffraction pattern has a 2θ value of 5.8±0.2, 6.3±0.2, 10.8±0.2, 11.8±0.2 , 13.8±0.2, and 17.5±0.2 have obvious characteristic absorption peaks.

The present invention also adopts thermogravimetric analysis method to study and characterize the crystalline form of aconitine C. The detection conditions are: starting from room temperature, heating gradient: heating up to 400°C at a rate of 10°C/min, and the protective gas is nitrogen.

The thermogravimetric analysis curve of the substantially pure crystalline form of aconitine C provided by the present invention is shown in Figure 2, which has the following characteristics: the sample TGA loses 15.93% of its weight, and when heated to 60°C, its weight loss is 8.58%; At 120°C, the weight loss is 7.35%.

The present invention also adopts differential scanning calorimetry to study and characterize the crystal form of aconitine C. The detection method is starting from 25°C, with a heating gradient: heating up to 280°C at a rate of 10°C/min, and the protective gas is nitrogen.

The differential scanning calorimetry curve of the substantially pure crystalline form of aconitine C provided by the present invention is shown in Fig. 2, and it has the following characteristics: the differential scanning calorimetry curve is at 67.1℃ and 81.6 respectively. Endothermic signals appeared at ℃, 123.7℃, and 173.8℃.

It is worth noting that for the X-ray powder diffraction pattern of the above crystal form, the characteristic peaks of the X-ray powder diffraction pattern may be between one machine and another machine and between one sample and another sample. There will be slight changes. The value may differ by about 1 unit, or by about 0.8 unit, or by about 0.5 unit, or by about 0.3 unit, or by about 0.1 unit, so the value given cannot be considered For absolute. Similarly, the values given in the differential scanning calorimetry graph of the above-mentioned crystal forms cannot be regarded as absolute.

The crystal form can also be characterized by other analytical techniques known in the art. For example, proton nuclear magnetic resonance spectroscopy ( ¹ HNMR).

The substantially pure crystalline form of aconitine C provided by the present invention has a hydrogen nuclear magnetic resonance spectrum as shown in FIG. 3.

The invention also provides a method for preparing the high-purity aconitine C crystal form.

The preparation method of the crystalline form of aconitine C provided by the present invention is obtained by using DMF as a solvent and a gas-solid permeation method. The specific operation is as follows: put aconitine A in a vial, take another larger bottle and add DMF to it, put the vial open in the larger bottle, seal it and let it stand at room temperature, and collect the solid.

In the preparation method of the crystalline form of aconitine C of the present invention, the vial is placed in a slightly larger bottle with an open mouth, and the time of standing at room temperature after sealing is not less than 2 days.

The preparation method of the crystalline form of aconitine C of the present invention has a crystal form content of more than 99%, high purity, consistent X-ray powder diffraction spectrum characteristics and DSC characteristics, stable properties, and good stability to light, humidity and heat .

The present invention also provides the application of the crystal form of aconitine C in the preparation of drugs for the prevention and/or treatment of rheumatoid arthritis RA, osteoarthritis, myofibritis, neck and shoulder pain, low back pain or cancer pain.

It can be known from the above technical scheme that the present invention discloses the preparation method of the aconitine C crystal form and the crystalline form of aconitine C. The X-ray powder diffraction spectrum of the crystal form of the present invention measured by using Cu-Kα rays is shown in Figure 1. The crystalline form of Caconitine A is obtained by gas-solid permeation method with DMF as solvent. The preparation process is simple and the obtained crystalline form has high purity. It is determined to be crystalline form C by XRD, DSC, TGA, ¹ HNMR characterization. The obtained aconitine crystal is a solvate crystal form, and the stability test shows that the crystal has good stability to light, humidity and heat.

Description of the drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art.

Figure 1 XRPD pattern of crystal form C;

Figure 2 TGA/DSC chart of crystal form C;

Figure 3 ¹ HNMR spectrum of Form C.

detailed description

The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

In order to further understand the present invention, the present invention will be described in detail below in conjunction with specific embodiments. In the following examples, unless otherwise specified, the test method is usually implemented under conventional conditions or conditions recommended by the manufacturer. All reagents are equally divided into analytical grade.

Crystal type identification test parameters:

The XRPD images were collected on PANalytacal Empyrean and X’Pert3 X-ray powder diffraction analyzers. The scanning parameters are shown in Table 1.

Table 1 XRPD test parameters

Thermogravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC)

TGA and DSC graphs were collected on TA Q5000 TGA/TA Discovery TGA5500 thermogravimetric analyzer and TA Q2000 DSC/TA Discovery DSC2500 differential scanning calorimeter respectively. Table 2 lists the test parameters.

Table 2 Test parameters of TGA and DSC

parameter	TGA	DSC
method	Linear heating	Linear heating
Sample tray	Aluminum pan, open	Aluminum plate, gland
temperature range	Room temperature-set endpoint temperature	25℃-set the end temperature
Scanning rate (℃/min)	10	10
Protective gas	Nitrogen	Nitrogen

Liquid NMR

The liquid NMR spectra were collected on a Bruker 400M NMR instrument with DMSO-d6 as the solvent.

Example 1. Preparation and identification of aconitine C crystal form

Obtained by gas-solid permeation method. The specific preparation steps are as follows: weigh 14.7 mg of aconitine in a 3 ml vial, take another 20 ml vial and add 2 ml DMF to it, and place the 3 ml vial open in 20 ml After sealing the vial, let it stand at room temperature. After about 6 days, collect the solid and perform XRPD, TGA/DSC and ¹ HNMR tests.

XRPD results show that there are obvious characteristic absorption peaks at diffraction angles (2θ angles) of 5.8±0.2, 6.3±0.2, 10.8±0.2, 11.8±0.2, 13.8±0.2, and 17.5±0.2. The TGA/DSC results show that when the sample is heated to 60°C, the weight loss is 8.5%; multiple endothermic signals appear on the DSC curve. ¹ HNMR results indicated that 1.68 moles of DMF (16.02 wt%) were detected in the sample.

It was identified as crystal form C, solvate.

The spectra are shown in Fig. 1 X-ray powder diffraction pattern of Aconitine C crystal form, Fig. 2 TGA/DSC analysis chart of Aconitine C crystal form, and Fig. 3 ¹ HNMR chart of Aconitine C crystal form.

Example 2. Preparation of aconitine C crystal form

Obtained by gas-solid permeation method. The specific preparation steps are as follows: weigh 14.7 mg of aconitine in a 3 ml vial, take another 20 ml vial and add 2 ml DMF to it, and place the 3 ml vial open in 20 ml The vial was sealed and allowed to stand at room temperature. After 2 days, the solid was collected and subjected to XRPD test. The result was consistent with Figure 1.

Example 3 Preparation of aconitine C crystal form

Obtained by gas-solid permeation method. The specific preparation steps are as follows: weigh 14.7 mg of aconitine in a 3 ml vial, take another 20 ml vial and add 2 ml DMF to it, and place the 3 ml vial open in 20 ml The vial was sealed and allowed to stand at room temperature. After 20 days, the solid was collected and subjected to XRPD test. The result was consistent with Figure 1.

Example 4 Stability test of aconitine C crystal form

In order to evaluate the solid-state stability of the crystal form, a suitable amount of samples were respectively weighed and left open for 1 week and 1 month at 25°C/60%RH and 40°C/75%RH, and sealed at 80°C for 24 hours. XRPD and HPLC characterization of the placed samples were performed to detect the crystal form changes and chemical purity.

The HPLC results are shown in Table 3. The results show that the chemical purity of the sample has hardly changed under the selected test conditions; the XRPD results show that the crystal form of the sample has not changed under the selected test conditions.

Table 3 Summary of stability data of crystal form C

In conclusion, the crystal form C has good physical and chemical stability.

Claims (8)

1. The crystalline form of aconitine C is characterized in that its X-ray powder diffraction pattern has obvious characteristics at 2θ values of 5.8±0.2, 6.3±0.2, 10.8±0.2, 11.8±0.2, 13.8±0.2, and 17.5±0.2 Absorption peak.
2. The crystalline form according to claim 1, wherein the thermogravimetric analysis curve has a weight loss of 8.58% when heated to 60°C, and a weight loss of 7.35% when heated to 120°C.
3. The crystal form according to claim 1, characterized in that the differential scanning calorimetry analysis curve has endothermic signals at 67.1°C, 81.6°C, 123.7°C, and 173.8°C, respectively.
4. The crystal form according to claim 1, wherein the proton nuclear magnetic resonance spectrum is shown in FIG. 3.
5. The preparation method of aconitine C crystal form according to claim 1, characterized in that DMF is used as a solvent and obtained by a gas-solid permeation method.
6. The preparation method according to claim 5, characterized in that the gas-solid permeation method specifically includes placing aconitine in a vial, taking a slightly larger bottle and adding DMF to it, and placing the vial open. In a slightly larger bottle, seal it and let it stand at room temperature to collect the solid.
7. The preparation method according to claim 6, characterized in that, the vial is placed in a slightly larger bottle with an open mouth and sealed and then left standing at room temperature for no less than 2 days.
8. The use of the crystalline form of aconitine C of claim 1 in the preparation of drugs for the prevention and/or treatment of rheumatoid arthritis RA, osteoarthritis, myofibritis, neck and shoulder pain, low back pain or cancer pain.

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