Acetyl L-carnitine hydrochloride derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medical compounds, and relates to an acetyl L-carnitine hydrochloride derivative as well as a preparation method and application thereof, in particular to an acetyl L-carnitine hydrochloride derivative which is synthesized by taking L-carnitine as a raw material through a one-step chemical reaction, and finally, the acetyl L-carnitine hydrochloride derivative is obtained through preparative chromatography purification. The high-purity acetyl L-carnitine hydrochloride derivative is obtained. The method for preparing the acetyl L-carnitine hydrochloride derivative provided by the invention is simple and convenient to operate and low in cost, amplification preparation can be performed as required, a new reference substance is provided for detection of the acetyl L-carnitine hydrochloride derivative in acetyl L-carnitine hydrochloride, and a guarantee is provided for further improvement of the quality of acetyl L-carnitine hydrochloride.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical compounds, and specifically relates to an acetyl-L-carnitine hydrochloride derivative and a preparation method and application thereof. Background Art

[0002] Acetyl-L-carnitine hydrochloride, as a naturally occurring substance in the body, participates in a series of important metabolic processes in the human body and has the effects of protecting nerves and promoting nerve function. It can be used to relieve paresthesia caused by diabetic peripheral neuropathy, assist in the treatment of progressive brain degeneration, Alzheimer's disease, etc., and is therefore widely used in food and medicine.

[0003] The study of acetyl-L-carnitine hydrochloride impurities is the premise for ensuring product quality and safety. Therefore, it is very necessary to study and control the impurities in acetyl-L-carnitine hydrochloride. It is not only the key to improve the quality of acetyl-L-carnitine hydrochloride, but also a reliable control for the production process of acetyl-L-carnitine hydrochloride, which has very important practical significance. In the process of studying the impurities of acetyl-L-carnitine hydrochloride series, our company discovered a potential new acetyl-L-carnitine hydrochloride derivative, which has not been found in any literature or patent reports. Summary of the invention

[0004] In order to solve the technical problems existing in the prior art, the purpose of the present invention is to provide an acetyl-L-carnitine hydrochloride derivative and a preparation method and application thereof.

[0005] In order to achieve the above purpose and the above technical effect, the technical solution adopted by the present invention is:

[0006] An acetyl-L-carnitine hydrochloride derivative, the structure of the acetyl-L-carnitine hydrochloride derivative is as follows:

[0007]

[0008] Further, the acetyl-L-carnitine hydrochloride derivative is prepared by the following steps:

[0009] Dissolve acetyl-L-carnitine hydrochloride and L-carnitine in acetic acid, add a condensation reagent dropwise at 70-100° C., and perform a chemical reaction to prepare an acetyl-L-carnitine hydrochloride derivative solution;

[0010] Separating the acetyl-L-carnitine hydrochloride derivative solution obtained in step (1) to obtain an acetyl-L-carnitine hydrochloride derivative preparation solution;

[0011] The prepared solution obtained in step (2) is concentrated to dryness, and then loaded onto a preparative chromatographic column for separation and purification to obtain a prepared solution containing an acetyl-L-carnitine hydrochloride derivative;

[0012] The prepared solution obtained in step (3) is concentrated to dryness to obtain a solid acetyl-L-carnitine hydrochloride derivative.

[0013] Furthermore, in the step (1), the molar ratio of acetyl-L-carnitine hydrochloride to L-carnitine is 1:1.0-30; preferably, the molar ratio of acetyl-L-carnitine hydrochloride to L-carnitine is 1:1.0-1.5.

[0014] Furthermore, in the step (1), the mass ratio of acetyl-L-carnitine hydrochloride to acetic acid is 1:1.0-3.0; preferably, the mass ratio of acetyl-L-carnitine hydrochloride to acetic acid is 1:1.2-1.8.

[0015] Furthermore, in the step (1), the reaction temperature is 20 to 100°C, preferably, the reaction temperature is 60 to 80°C.

[0016] Furthermore, in the step (1), the reaction time is 1 to 4 hours; preferably, the reaction time is 2 hours.

[0017] Furthermore, in step (1), the condensation reagent is acetyl chloride, acetic anhydride, oxalyl chloride, or thionyl chloride; preferably, the condensation reagent is acetyl chloride.

[0018] Furthermore, in the step (1), the molar ratio of acetyl-L-carnitine hydrochloride to the condensation reagent is 1:1.0-3.0; preferably, the molar ratio of acetyl-L-carnitine hydrochloride to the condensation reagent is 1:1.3.

[0019] Furthermore, in the step (2), the acetyl-L-carnitine hydrochloride derivative solution obtained in the step (1) is subjected to high performance liquid chromatography preparative separation, and the high performance liquid chromatography column is selected from: ThermoAPS-2Hypersil TM The specification of the HPLC column is 4.6x250mm; the chromatographic conditions also include: flow rate 0.5-1.5ml / min; the preferred flow rate is: 1.0ml / min; detection wavelength: 205nm.

[0020] Furthermore, in the step (3), the acetyl-L-carnitine hydrochloride derivative solution obtained in the step (2) is subjected to high performance liquid chromatography preparative separation, and the high performance liquid chromatography column is selected from: Agilent preparative liquid chromatography column XRsDiphenyl reverse phase preparative column; the specification of the high performance liquid chromatography column is: 21.2*250mm; the chromatographic conditions also include: flow rate 2~30mL / min; the preferred flow rate is: 5~10mL / min; detection wavelength: 205nm.

[0021] The invention also discloses an application of the acetyl-L-carnitine hydrochloride derivative in the purity detection of acetyl-L-carnitine hydrochloride.

[0022] The invention discloses an acetyl-L-carnitine hydrochloride derivative and a preparation method thereof, which are beneficial to the quality control of acetyl-L-carnitine hydrochloride and L-carnitine, thereby ensuring the use safety of acetyl-L-carnitine hydrochloride.

[0023] The invention provides a method for preparing acetyl-L-carnitine hydrochloride derivatives, obtains acetyl-L-carnitine hydrochloride derivatives with a chromatographic purity higher than 98%, and carries out structural identification of the acetyl-L-carnitine hydrochloride derivatives.

[0024] The acetyl-L-carnitine hydrochloride derivative prepared by the invention provides a new reference substance for detecting impurities in acetyl-L-carnitine hydrochloride, can calibrate the content of the acetyl-L-carnitine hydrochloride derivative in acetyl-L-carnitine hydrochloride, provides a direction for controlling the synthesis method of acetyl-L-carnitine hydrochloride, can also be used for qualitative and quantitative analysis of the acetyl-L-carnitine hydrochloride derivative in the production of acetyl-L-carnitine hydrochloride, is more conducive to the development of a detection method for related substances in acetyl-L-carnitine hydrochloride, thereby controlling product quality, has important application value for quality research and preparation research of acetyl-L-carnitine hydrochloride raw materials, and provides important guiding significance for safe use of medicines. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is the structural formula diagram of the acetyl-L-carnitine hydrochloride derivative of the present application;

[0026] Figure 2 This is a flow chart of the preparation principle of acetyl-L-carnitine hydrochloride derivatives of this application;

[0027] Figure 3 This is a typical HPLC chromatogram of the solution containing acetyl-L-carnitine hydrochloride derivative in Example 1;

[0028] Figure 4 This is a typical HPLC chromatogram of the solution containing acetyl-L-carnitine hydrochloride derivative of Example 2;

[0029] Figure 5 This is a typical HPLC chromatogram of the first purification preparation process of Example 3;

[0030] Figure 6 This is a typical HPLC chromatogram of the pure triacetyl-L-carnitine hydrochloride derivative of this embodiment;

[0031] Figure 7 It is the LC-MS spectrum of acetyl-L-carnitine hydrochloride derivative;

[0032] Figure 8 Acetyl L-carnitine hydrochloride derivative1 H-NMR spectrum;

[0033] Fig. 9 Acetyl L-carnitine hydrochloride derivative 13 C-NMR spectrum;

[0034] Fig.10 This is a typical HPLC chromatogram of the acetyl-L-carnitine hydrochloride raw material containing acetyl-L-carnitine hydrochloride derivatives. DETAILED DESCRIPTION

[0035] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without creative work belong to the scope of protection of the present invention. The experimental methods without specific conditions in the following embodiments are usually measured according to national standards. The experimental materials without the source in the following embodiments are all commercially available raw materials. The equipment used in each step in the following embodiments is conventional equipment. If there is no corresponding national standard, it is carried out according to the general international standards, conventional conditions, or according to the conditions recommended by the manufacturer. Unless otherwise specified, all parts are weight parts and all percentages are mass percentages. Unless otherwise defined or specified, all professional and scientific terms used in the present invention have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equal to the recorded content can be applied to the method of the present invention.

[0036] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments can be combined with each other. The present invention is further described below in conjunction with specific embodiments, but is not intended to be a limitation of the present invention.

[0037] The present invention discloses an acetyl-L-carnitine hydrochloride derivative, the structure of the acetyl-L-carnitine hydrochloride derivative is as follows: Figure 1 As shown:

[0038]

[0039] Further, the following steps are used to prepare the acetyl-L-carnitine hydrochloride derivative, as shown in the reaction formula: Figure 2 :

[0040] Dissolve acetyl-L-carnitine hydrochloride and L-carnitine in acetic acid, add a condensation reagent dropwise at 70-100° C., and perform a chemical reaction to prepare an acetyl-L-carnitine hydrochloride derivative solution;

[0041] Separating the acetyl-L-carnitine hydrochloride derivative solution obtained in step (1) to obtain an acetyl-L-carnitine hydrochloride derivative preparation solution;

[0042] The prepared solution obtained in step (2) is concentrated to dryness, and then loaded onto a preparative chromatographic column for separation and purification to obtain a prepared solution containing an acetyl-L-carnitine hydrochloride derivative;

[0043] The prepared solution obtained in step (3) is concentrated to dryness to obtain a solid acetyl-L-carnitine hydrochloride derivative.

[0044] Embodiment 1:

[0045] Preparation of solutions containing acetyl-L-carnitine hydrochloride derivatives

[0046] 20.0 g of L-carnitine, 29.7 g of acetyl-L-carnitine hydrochloride and 32.0 g of acetic acid were added to a 500 ml four-necked bottle, the temperature was raised to 70-80 ° C with stirring, and 12.6 g of acetyl chloride was added dropwise. After the addition was complete, the reaction was carried out at the same temperature for 2 h. After the reaction was completed, the solution was concentrated under reduced pressure to a certain volume, 34.0 g of acetone was added dropwise for crystallization, and suction was filtered to obtain 64.5 g of a solution containing an acetyl-L-carnitine hydrochloride derivative (the HPLC purity of the acetyl-L-carnitine hydrochloride derivative was 8.64%, see Figure 3 , see Table 1 for spectral data).

[0047] Table 1

[0048]

[0049]

[0050] Embodiment 2:

[0051] Preparation of crude acetyl-L-carnitine hydrochloride containing acetyl-L-carnitine hydrochloride derivatives

[0052] 20.0 g of L-carnitine, 29.7 g of acetyl-L-carnitine hydrochloride and 32.0 g of acetic acid were added to a 500 ml four-necked bottle, the temperature was raised to 70-80° C. with stirring, 20.3 g of oxalyl chloride was added dropwise, and the reaction was carried out at the same temperature for 2 h. After the reaction was completed, the solution was concentrated under reduced pressure to a certain volume, 34.0 g of acetone was added dropwise for crystallization, and suction was filtered to obtain 69.8 g of a solution containing an acetyl-L-carnitine hydrochloride derivative (the HPLC purity of the acetyl-L-carnitine hydrochloride derivative was 6.14%, see Figure 4 , see Table 2 for spectral data).

[0053] Table 2

[0054]

[0055]

[0056] Embodiment 3:

[0057] Purification and preparation of acetyl-L-carnitine hydrochloride derivatives:

[0058] Purification preparation process:

[0059] First purification: 30.00 g of the solution containing the acetyl-L-carnitine hydrochloride derivative prepared in Example 1 of the present application was loaded onto a high performance liquid chromatography column (chromatographic column: ThermoAPS-2Hypersil TM , 4.6x250mm) for purification and preparation, the mobile phase is a mixture of acetonitrile and 0.03mol / L potassium dihydrogen phosphate solution (the volume ratio of acetonitrile and potassium dihydrogen phosphate is 65:35, and the pH of the potassium dihydrogen phosphate solution is 4.7), the sample loading flow rate of the mobile phase is 1.0ml / min, the column temperature is 30°C, the detection wavelength is 205nm, and the preparation solution containing acetyl-L-carnitine hydrochloride derivative is collected and concentrated to dryness under reduced pressure to obtain 1.5g of solid crude acetyl-L-carnitine hydrochloride derivative (HPLC purity 44.5%, area normalization, see Figure 5 , the peak attribution is shown in Table 3).

[0060] Second purification: The above 1.2 g of the crude solid product of acetyl-L-carnitine hydrochloride derivative was loaded onto a high performance liquid chromatography column (chromatographic column is XRsDiphenyl reverse phase preparative column, 21.2x250 mm) for separation and purification in multiple times. The mobile phase is acetonitrile and water (the volume ratio of acetonitrile and water is 65:35). The loading flow rate of the mobile phase is 5.0 ml / min, the column temperature is 30°C, and the detection wavelength is 205 nm. The preparation solution containing the acetyl-L-carnitine hydrochloride derivative is collected and concentrated to dryness under reduced pressure to obtain 260 mg of the pure product of acetyl-L-carnitine hydrochloride derivative (HPLC purity 99.14%, area normalization, see Figure 6 , the peak attribution is shown in Table 4)

[0061] Table 3 HPLC chromatographic peak attribution and peak area

[0062]

[0063]

[0064] Table 4 HPLC chromatographic peak attribution and peak area

[0065]

[0066] Embodiment 4:

[0067] Structural confirmation of acetyl-L-carnitine hydrochloride derivatives: Mass spectrometry of acetyl-L-carnitine hydrochloride derivatives: ThermoScientificISQEM ultra-high performance liquid chromatography-mass spectrometry; ESI-: The peak of m / z 347.3 is the [M-Cl]- peak of acetyl-L-carnitine hydrochloride derivatives, obtained Figure 7 The liquid chromatography-mass spectrometry of the acetyl-L-carnitine hydrochloride derivative was determined to be 382.9.

[0068] NMR data of acetyl-L-carnitine hydrochloride derivatives: Instrument: BRUKERAV-600 NMR instrument, solvent: D2O; Determination data (the structural formula of acetyl-L-carnitine hydrochloride derivatives is shown in Figure 1 ) and then get Figure 8 Acetyl L-carnitine hydrochloride derivatives 1 H-NMR spectra and Fig. 9 Acetyl L-carnitine hydrochloride derivatives 13 C-NMR spectrum; Figure 8 and Fig. 9 The analysis is shown in the following table:

[0069] Table 5 Hydrogen spectrum analysis table

[0070] Serial number Chemical shift (ppm) Multiplicity J Number of protons 1 1.98 s 3H 2 2.54～2.59 d 2H 4 2.77～2.78 d 2H 6 3.03～3.05 d 18H 8 3.48～3.52 d 2H 10 3.75 s 2H 11 5.48 s 2H

[0071] Table 6 Carbon spectrum analysis table

[0072]

[0073]

[0074] Comprehensive analysis of all NMR spectrum data and mass spectrum splitting results indicates that the chemical name of the acetyl-L-carnitine hydrochloride derivative is (3R,3'R)-((3-acetoxy-4-(trimethylamino)butyryl)oxy)-4-(N,N,N-trimethylamino)butyrate, with the structural formula shown in Figure 1 .

[0075] Embodiment 5:

[0076] HPLC test results of acetyl-L-carnitine hydrochloride raw materials containing acetyl-L-carnitine hydrochloride derivatives: (HPLC chart of batch 2405946 is attached Fig.10 )

[0077] Table 7 HPLC chromatographic peak detection results (area normalization)

[0078]

[0079] In summary, the present invention provides a preparation method and application of acetyl-L-carnitine hydrochloride derivatives. In the process of producing acetyl-L-carnitine hydrochloride bulk drug, the present invention finds acetyl-L-carnitine hydrochloride derivatives, and confirms the structure of acetyl-L-carnitine hydrochloride derivatives through preparation, separation, purification and structural characterization. The structure of the acetyl-L-carnitine hydrochloride derivative is a new compound and has not been reported in the literature. At the same time, the present invention provides a method for preparing acetyl-L-carnitine hydrochloride derivatives, which can obtain high-purity acetyl-L-carnitine hydrochloride derivatives as reference substances or standard substances for controlling the content of acetyl-L-carnitine hydrochloride derivatives in acetyl-L-carnitine hydrochloride bulk drugs, and has a wide range of practical application value.

[0080] The above are only preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be regarded as limiting the present invention, and the protection scope of the present invention should be based on the scope defined by the claims. For ordinary technicians in this technical field, several improvements and modifications can be made without departing from the spirit and scope of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. An acetyl-L-carnitine hydrochloride derivative, characterized in that: The structure of the acetyl-L-carnitine hydrochloride derivative is as follows:

2. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 1, characterized in that: The method comprises the following steps: (1) dissolving acetyl-L-carnitine hydrochloride and L-carnitine in acetic acid, adding a condensation reagent dropwise at a certain temperature, and performing a chemical reaction to prepare an acetyl-L-carnitine hydrochloride derivative solution; (2) separating the acetyl-L-carnitine hydrochloride derivative solution obtained in step (1) to obtain an acetyl-L-carnitine hydrochloride derivative preparation solution; (3) concentrating the prepared solution obtained in step (2) to dryness, loading the sample onto a preparative chromatography column again, separating and purifying, and obtaining a prepared solution containing an acetyl-L-carnitine hydrochloride derivative; (4) Concentrating the preparation solution containing the acetyl-L-carnitine hydrochloride derivative obtained in step (3) to dryness to obtain a solid acetyl-L-carnitine hydrochloride derivative.

3. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 2, characterized in that: In the step (1), the molar ratio of acetyl-L-carnitine hydrochloride to L-carnitine is 1:1.0-3.

0.

4. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 2, characterized in that: In the step (1), the mass ratio of acetyl-L-carnitine hydrochloride to acetic acid is 1:1.0-3.

0.

5. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 2, characterized in that: In the step (1), the reaction temperature is 20 to 100°C.

6. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 2, characterized in that: In the step (1), the reaction time is 1 to 4 hours.

7. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 2, characterized in that: In the step (1), the condensation reagent is one or more of acetyl chloride, acetic anhydride, oxalyl chloride, and thionyl chloride.

8. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 2, characterized in that: In the step (1), the molar ratio of acetyl-L-carnitine hydrochloride to the condensation reagent is 1:1.0-3.

0.

9. The method for preparing the acetyl-L-carnitine hydrochloride derivative according to claim 2, characterized in that: In the step (2), the acetyl-L-carnitine hydrochloride derivative solution obtained in the step (1) is subjected to high performance liquid chromatography preparative separation, and the high performance liquid chromatography column is selected from: ThermoAPS-2Hypersil TM The specification of the high performance liquid chromatography column is 4.6*250mm; the chromatographic conditions also include: a flow rate of 0.5-1.5ml / min.

10. Use of the acetyl-L-carnitine hydrochloride derivative according to claim 1 in the purity detection of acetyl-L-carnitine hydrochloride.

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