Preparation method and application of 7-hydroxy anisodamine

Abstract

The invention relates to the field of pharmaceutical chemicals, in particular to a preparation method and application of 7-hydroxy anisodamine. According to the preparation method of the 7-hydroxyanisodamine, provided by the invention, a qualified 7-hydroxyanisodamine reference substance is provided for research on impurities of an anisodamine hydrobromide bulk drug and a preparation thereof, and the 7-hydroxyanisodamine reference substance is high in purity and can be used as a reference substance to be applied to quality control of the anisodamine hydrobromide bulk drug and the preparation thereof; the quality control of the anisodamine hydrobromide raw material medicine and the preparation thereof is favorably improved, and the medicine safety is improved; and in addition, the unqualified anisodamine hydrobromide raw material medicine and the preparation thereof can be treated in time, so that the storage cost is reduced.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and specifically to a preparation method and application of 7-hydroxy anisodamine. Background Art

[0002] Anisodamine hydrobromide, whose structural formula is This product is an anticholinergic drug that blocks M cholinergic receptors, has the effects of relaxing smooth muscle, relieving vasospasm, improving microcirculation, and has an analgesic effect. However, its effects of dilating pupils and inhibiting glandular secretion are weak, and it rarely causes central excitatory symptoms.

[0003] Impurities with the structure of may be generated during the production and storage of anisodamine hydrobromide raw materials and their preparations. The generation mechanism is that anisodamine hydrobromide raw materials and their preparations decompose anisodamine, and then anisodamine (C 17 H 23 NO4, with a molecular weight of 305.37) undergoes hydroxylation to produce this impurity. This impurity is called 7-hydroxy anisodamine. After detection, its molecular formula is C 17 H 23 NO5, with a molecular weight of 321.37, and its chemical name is (1R,3r,5S,6S,7R)-6,7-dihydroxy-8-methyl-1-8-azabicyclo[3.2.1]octan-3-yl-(S)-3-hydroxy-2-phenylpropionate. The chemical equation of the generation mechanism of this impurity is as follows:

[0004]

[0005] Currently, there is no high-purity preparation method for the above substance, so it is impossible to obtain a qualified 7-hydroxy anisodamine reference substance for quality control. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a preparation method and application of 7-hydroxy anisodamine. The preparation method provided by the present invention can prepare 7-hydroxy anisodamine with high purity, which can be used as a reference substance in the quality control of anisodamine hydrobromide raw materials and their preparations.

[0007] The present invention provides a preparation method of 7-hydroxy anisodamine, including the following steps:

[0008] S1) Extract and purify the anisodamine extract with dichloromethane and sulfuric acid;

[0009] S2) Perform column chromatography on the liquid obtained in step S1);

[0010] The specific column chromatography is as follows: using reverse-phase silica gel as the stationary phase, using a mixed solution of acetonitrile and phosphoric acid as the mobile phase, and the elution program is:

[0011]

[0012] S3) Crystallize and dry the liquid obtained in step S2) together with petroleum ether and acetone to obtain 7-hydroxyanisodamine.

[0013] In the present invention, the anisodamine extract is first extracted and purified with dichloromethane and sulfuric acid. Specifically, after concentrating the anisodamine extract to obtain a concentrated solution of the extract, adjust the pH to 8.0 - 10.0, then extract with dichloromethane and sulfuric acid to obtain an extract solution. Mix the extract solution with sulfuric acid, separate the aqueous phase, and concentrate the obtained aqueous phase. In certain embodiments of the present invention, add ethanol to the anisodamine extract and perform vacuum concentration at 70°C - 90°C to obtain a concentrated solution of the extract, then adjust the pH to 8.0 - 10.0 with a NaOH solution having a mass concentration of 8% - 12%, and then extract 2 - 4 times with dichloromethane having a volume 0.8 - 1.2 times that of the material to be extracted and purified to obtain an extract solution. Mix the extract solution with sulfuric acid having a volume concentration of 1.5% - 2.5%, separate the aqueous phase, and perform vacuum concentration on the obtained aqueous phase solution at 70°C - 90°C to obtain a sample loading solution. The volume concentration of the sulfuric acid in the present invention is 1.5% - 2.5%; the volume ratio of dichloromethane to sulfuric acid is 1:(0.4 - 0.6). Based on the calculation of 20 unit volumes of the concentrated solution of the extract, mix the extract solution with 8 - 12 unit volumes of sulfuric acid.

[0014] The anisodamine extract in the present invention is specifically a hydrochloric acid extract of anisodamine. Specifically, the anisodamine extract is obtained by extraction with hydrochloric acid having a molar concentration of 4% - 6%. In certain embodiments of the present invention, soak anisodamine in hydrochloric acid having a molar concentration of 4% - 6% for extraction, filter, and then perform vacuum concentration at 70°C - 90°C to obtain the anisodamine extract.

[0015] After the anisodamine extract is extracted and purified with dichloromethane and sulfuric acid in the present invention, perform column chromatography on the liquid obtained in step S1). Specifically, perform column chromatography on the obtained liquid, enrich and concentrate the chromatographed solution obtained after column chromatography, then adjust the pH to 9.0 - 10.0, and then perform extraction and purification with dichloromethane and then concentrate. In certain embodiments of the present invention, perform column chromatography on the liquid obtained in step S1), perform column chromatography again on the chromatographed solution obtained after column chromatography to enrich the target component, then perform vacuum concentration on the enriched material at 70°C - 90°C, then adjust the pH to 9.0 - 10.0 with a NaOH solution having a mass concentration of 8% - 12%, then perform extraction and purification with dichloromethane, dehydrate and filter, and then perform vacuum concentration at 40°C - 60°C to obtain a paste.

[0016] The dosage of dichloromethane in the present invention is 0.8 to 1.2 times the volume of the material to be extracted and purified, preferably 1 time. The number of extraction times in the present invention is 4 to 6 times. The present invention uses anhydrous sodium sulfate for dehydration. The column chromatography in the present invention preferably uses reversed-phase bonded silica gel as the stationary phase and a mixed solution of acetonitrile and phosphoric acid with a volume concentration of 0.1 wt% to 0.3 wt% as the mobile phase.

[0017] After the column chromatography in the present invention, the liquid obtained in step S2), petroleum ether and acetone are crystallized and dried together to obtain 7-hydroxyanisodamine. Specifically, a mixed solution of petroleum ether and acetone is added to the liquid obtained in step S2), heated and dissolved at 45°C to 55°C, stirred and crystallized at 0°C to 10°C, and then the obtained crystals are dried under reduced pressure at 40°C to 60°C to obtain the anisodamine hydrobromide impurity compound. The volume ratio of the petroleum ether and acetone in the present invention is 1:(0.9 to 1.1), and the total dosage of the petroleum ether and acetone is 2 to 3 times the volume of the material obtained in step S2), preferably 2.5 times.

[0018] The present invention also provides the application of the above-mentioned anisodamine hydrobromide impurity compound as a reference substance in the impurity detection of anisodamine hydrobromide raw material medicine and its preparations. The preparation method provided by this patent provides a qualified 7-hydroxyanisodamine reference substance for the impurity research of anisodamine hydrobromide raw material medicine and its preparations, which is beneficial to improving the quality control of anisodamine hydrobromide raw material medicine and its preparations and enhancing the drug safety.

[0019] The present invention provides a preparation method and application of 7-hydroxyanisodamine. The purification preparation method of 7-hydroxyanisodamine provided by the present invention has not been reported. The preparation method of 7-hydroxyanisodamine provided by the present invention provides a qualified 7-hydroxyanisodamine reference substance for the impurity research of anisodamine hydrobromide raw material medicine and its preparations, which is beneficial to improving the quality control of anisodamine hydrobromide raw material medicine and its preparations and enhancing the drug safety; and it also helps to timely process the substandard anisodamine hydrobromide raw material medicine and its preparations, reducing the storage cost. Description of the Drawings

[0020] Figure 1 It is the preparation flow chart of 7-hydroxyanisodamine described in the present invention;

[0021] Figure 2 It is the liquid chromatogram of 7-hydroxyanisodamine obtained in Example 1 of the present invention;

[0022] Figure 3 It is the mass spectrum of 7-hydroxyanisodamine obtained in Example 1 of the present invention;

[0023] Figure 41H NMR spectrum of 7-hydroxyanisodamine obtained in Example 1 of the present invention;

[0024] Figure 5 13C NMR spectrum of 7-hydroxyanisodamine obtained in Example 1 of the present invention. Detailed implementation manners

[0025] The present invention discloses a preparation method and application of 7-hydroxyanisodamine. Those skilled in the art can draw on the content of this article and appropriately improve the process parameters to achieve. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all regarded as included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and those skilled in the art can obviously make changes or appropriate alterations and combinations to the methods and applications in this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0026] As Figure 1 shown, Figure 1 is the preparation flow chart of 7-hydroxyanisodamine described in the present invention. The present invention prepares the anisodamine hydrobromide impurity compound described in the present invention according to the flow Figure 1 shown.

[0027] The present invention is further described below in conjunction with embodiments:

[0028] Example 1

[0029] (1) Extraction

[0030] 50 kg of anisodamine medicinal materials are pulverized, added with 600 L of 5% hydrochloric acid solution for soaking extraction, filtered to obtain an extraction solution, and concentrated under reduced pressure at 80 °C to 50 L. 200 L of absolute ethanol is added to the extraction solution, and it is left standing for 12 h, filtered, the filtrate is concentrated under reduced pressure at 80 °C to 20 L, the pH is adjusted to 9.5 with 10% NaOH solution, and it is extracted 3 times with dichloromethane in a ratio of 1:1, and the dichloromethane extraction solutions are combined; 10 L of 2% sulfuric acid is added to the dichloromethane extraction solution, mixed and left standing, the aqueous phase is separated, concentrated under reduced pressure at 80 °C to 2 L, and filtered with a 0.22 μm filter cloth to obtain a sample loading solution.

[0031] (2) Column chromatography purification

[0032] The sample loading solution in (1) is loaded onto a reverse-phase silica gel column (10 μm, reverse-phase bonded silica gel), and eluted with acetonitrile - 0.2% phosphoric acid. The elution program is as follows. The components containing the target substance are collected, loaded onto the column for purification again, eluted in the same way, and the target components are enriched again, and concentrated under reduced pressure at 80 °C to obtain 30 mL of a concentrated solution.

[0033]

[0034] (3) Extraction purification

[0035] Adjust the pH of the concentrated solution in (2) to 9.5 with 10% NaOH solution, extract it 5 times with dichloromethane at a ratio of 1:1, combine the extracts, add anhydrous sodium sulfate for dehydration and filtration, and concentrate it under reduced pressure to a paste at 50 °C.

[0036] (4) Crystallization and drying

[0037] Add 2 mL of petroleum ether - acetone (volume ratio 1:1), heat and dissolve it at 50 °C, stir it at 5 °C, precipitate crystals, and dry it under reduced pressure at 50 °C to obtain 0.62 g of the target product. Chromatographic purity: 98.39%. The chromatogram is as Figure 2 shown. Figure 2 This is the liquid chromatogram of 7 - hydroxyscopolamine obtained in Example 1 of the present invention. Figure 2 The corresponding data is as described in Table 1:

[0038] Table 1

[0039] Peak No. Compound Name Retention Time Area Height Area % Theoretical Plate Number Resolution Tailing Factor 1 4.598 11366 1370 0.10 6827 -- 1.32 2 5.128 15946 1739 0.15 6837 2.25 1.29 3 7.941 29278 2326 0.27 8850 9.60 1.18 4 9.455 30304 2093 0.28 9748 4.20 1.16 5 10.078 24927 1671 0.23 10147 1.59 1.13 6 7-Hydroxyanisodamine 11.584 10743340 486549 98.39 6579 3.10 1.78 7 33.030 28311 2531 0.26 190340 49.07 1.59 8 36.930 35900 4038 0.33 371281 14.31 1.33 Total 10919372 502316 100.00

[0040] (5) Structure confirmation

[0041] Mass spectrometry: As Figure 3 shown. Figure 3 This is the mass spectrum of 7 - hydroxyscopolamine obtained in Example 1 of the present invention. It can be Figure 3 seen that the theoretical molecular weight is 321.37, and the measured molecular ion peaks are m / z 322.2 [M + 1] and m / z 322.3 [M + 2], which are consistent with the theoretical molecular weight.

[0042] Hydrogen spectrum: As Figure 4 described. Figure 4 This is the hydrogen spectrum of 7 - hydroxyscopolamine obtained in Example 1 of the present invention, and the corresponding data is 1 H NMR(400MHz, Methanol - d4)δ7.38 - 7.26(m, 5H), 4.97(t, J = 5.4Hz, 1H), 4.37(d, J = 6.4Hz, 1H), 4.16 - 4.09(m, 1H), 3.97(d, J = 6.4Hz, 1H), 3.75(qd, J = 5.5, 2.7Hz, 2H), 2.99(p, J = 1.8Hz, 1H), 2.90(dt, J = 3.9, 1.8Hz,1H), 2.47(s,3H), 2.16(dddd,J = 29.5,15.7,5.5,4.1Hz, 2H), 1.59(dt,J = 15.3,2.3Hz, 1H), 1.40–1.33(m, 1H).

[0043] Carbon spectrum: As Figure 5 described. Figure 5This is the carbon spectrum of 7-hydroxyanisodamine obtained in Example 1 of the present invention, and the corresponding data are 13 C NMR(101MHz,Methanol-d4)δ173.07,137.22,129.98,129.98,129.17,129.17,128.91,75.18,75.04,68.73,67.59,67.49,64.7,56.19,36.51,29.11,28.94。

[0044] The above are only the preferred specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, making equivalent substitutions or changes should be covered within the protection scope of the present invention.

Claims

1. Preparation method of 17-hydroxyanisodamine, characterized in that, It includes the following steps: S1) Extract and purify the anisodamine extract with dichloromethane and sulfuric acid; S2) Perform column chromatography on the material obtained in step S1); The specific column chromatography is as follows: Use reverse-phase silica gel as the stationary phase, and the elution program is: S3) Crystallize and dry the material obtained in step S2) together with petroleum ether and acetone to obtain 7-hydroxyanisodamine.

2. The preparation method according to claim 1, characterized in that, In step S1), the anisodamine extract is the hydrochloric acid extract of anisodamine.

3. The preparation method according to claim 2, characterized in that, In step S1), the anisodamine extract is obtained by extraction with hydrochloric acid with a molar concentration of 4% - 6%.

4. The preparation method according to claim 1, characterized in that, Step S1) is specifically: Concentrate the anisodamine extract to obtain a concentrated extract, then adjust the pH to 8.0 - 10.0, then extract with dichloromethane to obtain an extract, mix the extract with sulfuric acid, separate the aqueous phase, and concentrate the obtained aqueous phase.

5. The preparation method according to claim 4, characterized in that, In step S1), calculated based on 20 unit volumes of the concentrated extract of the extract, mix the extract with 8 - 12 unit volumes of sulfuric acid.

6. The preparation method according to claim 4, wherein, In step S1), the volume concentration of the sulfuric acid is 1.5% - 2.5%.

7. The preparation method according to claim 2, characterized in that, In step S2), use a mixed solution of acetonitrile and 0.1wt% - 0.3wt% phosphoric acid as the mobile phase.

8. The preparation method according to claim 2, characterized in that, Step S2) also includes: Enrich and concentrate the chromatographed solution obtained after column chromatography, then adjust the pH to 9.0 - 10.0, then extract and purify with dichloromethane and then concentrate.

9. The preparation method according to claim 2, wherein, In step S3), the volume ratio of the petroleum ether to the acetone is 1:(0.9 - 1.1), and the total amount of the petroleum ether and the acetone used is 2 - 3 times the volume of the material obtained in step S2) by weight.

10. The application of 7-hydroxyanisodamine obtained by the preparation method according to any one of claims 1 - 9 as a reference substance in the impurity detection of anisodamine hydrobromide bulk drug and its preparations.

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