Preparation method and application of 2 '-hydroxyanisodamine
Through dichloromethane and sulfuric acid extraction, reverse phase silica gel column chromatography and crystallization drying, the high-purity preparation problem of 2’-hydroxyanisopamine impurities in the raw materials for anisopamine hydrobromide was solved, and the preparation of reference products was realized, improving quality control and drug safety.
Patent Information
- Application Number
- CN202510136849.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-07
- Publication Date
- 2025-07-04
AI Technical Summary
In the prior art, the 2’-hydroxyanisoposamine impurities produced by the hydrobromide raw materials and their preparations during production and storage lack high-purity preparation methods, resulting in the inability to effectively control.
2’-hydroxyanisoposamine was prepared by extraction purification of dichloromethane and sulfuric acid, reverse phase silica gel column chromatography and crystallization drying. High purity of 2’-hydroxyanisoposamine was obtained by extraction, elution and crystallization steps.
It provides a high-purity 2’-hydroxyanisopsamine reference product for the quality control of the anisopsamine hydrobromide raw materials and their preparations, improves the safety of drugs and reduces the storage cost of unqualified products.
Smart Images
Figure BDA0005263533340000021 
Figure BDA0005263533340000022 
Figure FDA0005263533330000011
Abstract
Description
Technical Field
[0001] The present invention relates to the field of pharmaceutical chemistry, and specifically to a preparation method and application of 2'-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 muscles, 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 is decomposed from anisodamine hydrobromide raw materials and their preparations, and then anisodamine (C 17 H 23 NO4, molecular weight 305.37) is oxidized to produce this impurity. This impurity is called 2'-hydroxy anisodamine, whose molecular formula is C 17 H 23 NO5, molecular weight 321.37, and chemical name is (1R,3R,5R,6S)-6-hydroxy-8-methyl-8-azabicyclo[3.2.1]octan-3-yl (R)-2,3-dihydroxy-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 2'-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 2'-hydroxy anisodamine. The preparation method provided by the present invention can prepare 2'-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 2'-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: 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 dichloromethane to obtain 2'-hydroxyanisodamine.
[0013] In the present invention, the anisodamine extract is first extracted and purified with dichloromethane and sulfuric acid. Specifically, the anisodamine extract is concentrated to obtain a concentrated extract, and then the pH is adjusted to 8.0 - 10.0, and then extracted with dichloromethane to obtain an extract. The extract is mixed with sulfuric acid, and the aqueous phase is separated, and the obtained aqueous phase is concentrated. In certain embodiments of the present invention, the anisodamine extract is added with ethanol and concentrated under reduced pressure at 70°C - 90°C to obtain a concentrated extract, and then the pH is adjusted to 8.0 - 10.0 with an NaOH solution having a mass concentration of 8% - 12%, and then extracted 2 - 4 times with dichloromethane having a volume 0.8 times - 1.2 times that of the material to be extracted and purified to obtain an extract. The extract is mixed with sulfuric acid having a volume concentration of 1.5% - 2.5%, and the aqueous phase is separated, and the obtained aqueous phase is concentrated under reduced pressure at 70°C - 90°C to obtain a sample loading solution. Calculated based on 20 unit volumes of the concentrated extract of the extract, the extract is mixed 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, anisodamine is soaked in hydrochloric acid having a molar concentration of 4% - 6% for extraction, filtered, and then concentrated under reduced pressure 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, the material obtained in step S1) is subjected to column chromatography. Specifically, the obtained material is subjected to column chromatography, and the chromatographic solution obtained after column chromatography is enriched and concentrated, and then the pH is adjusted to 9.0 - 10.0, and then extracted and purified with dichloromethane and then concentrated. In certain embodiments of the present invention, the liquid obtained in step S1) is subjected to column chromatography, and the chromatographic solution obtained after column chromatography is subjected to column chromatography again to enrich the target component, and then the enriched material is concentrated under reduced pressure at 70°C - 90°C and then the pH is adjusted to 9.0 - 10.0 with an NaOH solution having a mass concentration of 8% - 12%, and then extracted and purified with dichloromethane, dehydrated and filtered, and then concentrated under reduced pressure 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 reverse-phase silica gel as the stationary phase and a mixed solution of acetonitrile and phosphoric acid with a volume concentration of 0.1% to 0.3% as the mobile phase.
[0017] After the column chromatography in the present invention, the material obtained in step S2) and dichloromethane are crystallized and dried together to obtain 2'-hydroxyanisodamine. Specifically, dichloromethane is added to the material obtained in step S2), heated and dissolved at 35°C to 45°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 2'-hydroxyanisodamine. The dosage of dichloromethane in the present invention is 2.5 to 3.5 times the volume of the material obtained in step S2), that is, 2.5 to 3.5 times the volume of the paste obtained in step S2).
[0018] The present invention also provides the application of 2'-hydroxyanisodamine obtained by the above preparation method as a reference substance in the impurity detection of anisodamine hydrobromide raw materials and their preparations. The preparation method provided by this patent provides a qualified 2'-hydroxyanisodamine reference substance for the impurity research of anisodamine hydrobromide raw materials and their preparations, which is beneficial to improving the quality control of anisodamine hydrobromide raw materials and their preparations and enhancing the drug safety.
[0019] The present invention provides a preparation method and application of 2'-hydroxyanisodamine. The preparation method of 2'-hydroxyanisodamine provided by the present invention has not been reported. The preparation method of 2'-hydroxyanisodamine provided by the present invention provides a qualified 2'-hydroxyanisodamine reference substance for the impurity research of anisodamine hydrobromide raw materials and their preparations, which is beneficial to improving the quality control of anisodamine hydrobromide raw materials and their preparations and enhancing the drug safety; and it also helps to timely process the substandard anisodamine hydrobromide raw materials and their preparations, reducing the storage cost. Description of the Drawings
[0020] Figure 1 It is the preparation flow chart of 2'-hydroxyanisodamine described in the present invention;
[0021] Figure 2 It is the liquid chromatogram of the refined product of 2'-hydroxyanisodamine obtained in Example 1 of the present invention;
[0022] Figure 3 It is the mass spectrum of 2'-hydroxyanisodamine obtained in Example 1 of the present invention;
[0023] Figure 4 It is the hydrogen spectrum of 2'-hydroxyanisodamine obtained in Example 1 of the present invention;
[0024] Figure 5 This is the carbon spectrum of 2'-hydroxyanisodamine obtained in Example 1 of the present invention. Detailed implementation manners
[0025] The present invention discloses a preparation method and application of 2'-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 This is the preparation flow chart of 2'-hydroxyanisodamine described in the present invention. The present invention prepares the anisodamine hydrobromide impurity compound described in the present invention according to the Figure 1 shown process.
[0027] The present invention is further elaborated 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 and 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, allowed to stand 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 extracted 3 times with dichloromethane at 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 allowed to stand, the aqueous phase is separated, concentrated under reduced pressure at 80 °C to 2 L, and filtered through 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), eluted with acetonitrile - 0.2% phosphoric acid, and the elution program is as follows. Collect the components containing the target substance, load them onto the column for purification again, elute in the same way, enrich the target components again, and concentrate under reduced pressure at 80 °C to obtain 30 mL of a concentrated solution.
[0033] Time min Mobile phase 0～65 0.1% Phosphoric acid - Acetonitrile [Volume ratio 95:5] 65～80 0.1% Phosphoric acid - Acetonitrile [Volume ratio 20:80]
[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 extraction solutions, add anhydrous sodium sulfate for dehydration and filtration, and concentrate it under reduced pressure at 50 °C to a paste.
[0036] (4) Crystallization and drying
[0037] Add 2.5 mL of dichloromethane, heat and dissolve it at 40 °C, stir it at 5 °C to precipitate crystals, and dry it under reduced pressure at 50 °C to obtain 0.45 g of the target product. Chromatographic purity: 95.44%. The chromatogram is as Figure 2 shown, Figure 2 the liquid chromatogram of 2'-hydroxyanisodamine obtained in Example 1 of the present invention, Figure 2 and the corresponding data are as described in Table 1:
[0038] Table 1
[0039] Peak number Compound name Retention time Area Height Area % Theoretical plate number Resolution Tailing factor 1 4.376 94202 11550 1.13 6415 -- 1.28 2 6.862 236870 17378 2.84 7314 9.22 1.14 3 2'-Hydroxyanisodamine 7.586 7959535 597721 95.44 7158 2.13 1.41 4 8.356 21335 1673 0.26 9263 2.18 -- 5 10.100 27732 1539 0.33 6427 4.10 1.07 Total 8339675 629861 100.00
[0040] (5) Structure confirmation
[0041] Mass spectrum: As Figure 3 shown, Figure 3 the mass spectrum of 2'-hydroxyanisodamine 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] 1H NMR spectrum: As Figure 4 described, Figure 4 the 1H NMR spectrum of 2'-hydroxyanisodamine obtained in Example 1 of the present invention, and the corresponding data are 1 H NMR (400 MHz, Methanol-d4) δ 7.60–7.54 (m, 2H), 7.43–7.32 (m, 3H), 5.03 (t, J = 4.8 Hz, 1H), 4.63 (d, J = 7.5 Hz, 1H), 4.31 (d, J = 11.2 Hz, 1H), 3.91 (s, 1H), 3.80 (d, J = 11.2 Hz, 1H), 3.65 (s, 1H), 3.00 (s, 3H), 2.60 (t, J = 18.0 Hz, 2H), 2.41 (dd, J = 14.8, 7.9 Hz, 1H), 2.08 (d, J = 15.1 Hz, 2H), 1.87 (d, J = 16.1 Hz, 1H).
[0043] 13C NMR spectrum: As Figure 5 described, Figure 5 the 13C NMR spectrum of 2'-hydroxyanisodamine obtained in Example 1 of the present invention, and the corresponding data are 1313C NMR (101 MHz, Methanol-d4) δ 173.68, 140.57, 129.62, 129.62, 129.34, 129.34, 126.76, 80.77, 73.01, 71.62, 68.81, 67.35, 64.42, 41.39, 37.08, 35.11, 33.74。
[0044] As described above, it is only a preferred specific embodiment 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, shall be covered by the protection scope of the present invention.
Claims
1. A preparation method of 2'-hydroxyanisodamine, characterized in that, It includes the following steps: S1) Extract and purify the anisodamine extract with dichloromethane and sulfuric acid; S2) Subject the material obtained in step S1) to column chromatography; The specific column chromatography is as follows: using reverse-phase silica gel as the stationary phase, and a mixed solution of acetonitrile and phosphoric acid as the mobile phase. The elution program is: S3) Crystallize and dry the material obtained in step S2) together with dichloromethane to obtain 2'-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 having a molar concentration of 4% to 6%.
4. The preparation method according to claim 1, characterized in that, Step S1) specifically is: 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 1, characterized in that, In step S2), use a mixed solution of acetonitrile and phosphoric acid with a volume concentration of 0.1% - 0.3% as the mobile phase.
8. The preparation method according to claim 1, characterized in that, Step S2) also includes: Enrich and concentrate the material 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 1, characterized in that, In step S3), the dosage of dichloromethane is 2.5 - 3.5 times the volume of the material obtained in step S2) by weight.
10. The application of 2'-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.