Preparation method of Fmoc-Lys (Mtt)-OH
By reacting Fmoc-Lys·HCl with Mtt-Cl under the action of organic alkali, Fmoc-Lys(Mtt)-OH is prepared, which solves the problems of low yield and high cost in the prior art, and realizes an efficient and low-cost preparation method.
Patent Information
- Application Number
- CN202411626542.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-14
- Publication Date
- 2025-02-25
AI Technical Summary
The method for preparing Fmoc-Lys(Mtt)-OH in the prior art has low reaction yield, large amount of Mtt-Cl, high cost of raw and auxiliary materials, and is not suitable for modern industrial production.
Fmoc-Lys·HCl and Mtt-Cl were reacted under the action of organic alkali to obtain Fmoc-Lys(Mtt)-OH, which simplified the process, simple operation, mild reaction conditions, and reduced the difficulty of industrial production.
The high yield and high purity of Fmoc-Lys(Mtt)-OH are achieved, with a yield of more than 80% and a purity of more than 99%, reducing production costs.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of drug synthesis, and particularly relates to a method for preparing Fmoc-Lys(Mtt)-OH. Background Art
[0002] Fmoc-Lys(Mtt)-OH is a commonly used intermediate for the synthesis of peptides such as semaglutide. Its Chinese name is fluorenylmethoxycarbonyl-N'-4-methyltriphenyl-lysine, and its molecular formula is C 41 H 40 N2O4, CAS number is 167393-62-6, and its structural formula is as follows:
[0003]
[0004] In the prior art, a common method for preparing Fmoc-Lys(Mtt)-OH is: the raw materials Lys and Mtt-Cl are first synthesized into Mtt-Lys(Mtt)-OH, one of the Mtt is removed with an acid to obtain Lys(Mtt)-OH, and then Fmoc-Osu is used to protect the obtained Fmoc-Lys(Mtt)-OH. The reaction yield is low, the amount of Mtt-Cl used is large, the cost of raw materials is high, and it is not suitable for modern industrial production. The specific reaction is as follows:
[0005] Summary of the invention
[0006] In view of this, the object of the present invention is to provide a method for preparing Fmoc-Lys(Mtt)-OH. The preparation method provided in the present application has a simple process, mild reaction conditions, and the obtained product has a high yield and purity.
[0007] The present application provides a method for preparing Fmoc-Lys(Mtt)-OH, comprising the following steps:
[0008] The compound represented by formula (1) reacts with the compound represented by formula (II) under the action of an organic base to obtain Fmoc-Lys(Mtt)-OH represented by formula (III);
[0009]
[0010] In some specific implementations, the organic base is selected from at least one of triethylamine and N,N-diisopropylethylamine.
[0011] In some specific implementations, the molar ratio of the compound represented by formula (1), the compound represented by formula (II) and the organic base is 1:(0.8-1.3):(2-2.5).
[0012] In some specific implementations, the organic solvent for the reaction is selected from dichloromethane, methanol, ethanol or tetrahydrofuran.
[0013] In some specific implementations, the reaction temperature is 5°C to 25°C, and the reaction time is 2h to 10h.
[0014] In some specific implementations, the compound represented by formula (1) is mixed evenly with an organic solvent, and then the compound represented by formula (II) is added and mixed evenly, and then an organic base is added to react.
[0015] In some specific implementations, the temperature for adding the compound represented by formula (II) is 5°C to 25°C.
[0016] In some specific implementations, the organic base is added dropwise.
[0017] In some specific implementations, the obtained reaction product is further subjected to post-processing.
[0018] In some specific implementations, the post-processing includes:
[0019] The obtained reaction product is washed with citric acid solution and brine in turn, dried, filtered and then crystallized to obtain Fmoc-Lys(Mtt)-OH represented by formula (III).
[0020] The present application uses Fmoc-Lys·HCl shown in formula (I) as a starting material, and reacts with Mtt-Cl shown in formula (II) in the presence of an organic base to synthesize Fmoc-Lys(Mtt)-OH shown in formula II(I), which simplifies the process, is simple to operate, has mild reaction conditions, reduces the difficulty of industrial production, does not involve precious raw materials, is conducive to controlling production costs, and obtains Fmoc-Lys(Mtt)-OH with high purity and high yield. Experimental results show that the yield of Fmoc-Lys(Mtt)-OH prepared by the preparation method provided in the present application is more than 80%, and the purity is more than 99%. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 This is the high performance liquid chromatography (HPLC) spectrum of the product prepared in Example 1;
[0022] Figure 2 The infrared spectrum of the product prepared in Example 1;
[0023] Figure 3 This is the high performance liquid chromatography (HPLC) spectrum of the product prepared in Example 2;
[0024] Figure 4 The infrared spectrum of the product prepared in Example 2;
[0025] Figure 5 This is the high performance liquid chromatography (HPLC) spectrum of the product prepared in Example 3;
[0026] Figure 6 The infrared spectrum of the product prepared in Example 3;
[0027] Figure 7 This is the NMR spectrum of the product prepared in Example 3. DETAILED DESCRIPTION
[0028] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0029] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.
[0030] In addition, the terms "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. In the description of the present invention, "multiple" means two or more, unless otherwise clearly and specifically defined. In addition, the terms "installed", "connected" and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.
[0031] The present application provides a method for preparing Fmoc-Lys(Mtt)-OH, comprising the following steps:
[0032] The compound represented by formula (1) reacts with the compound represented by formula (II) under the action of an organic base to obtain Fmoc-Lys(Mtt)-OH represented by formula (III);
[0033]
[0034] The present application uses the compound represented by formula (1) and the compound represented by formula (II) as raw materials, wherein the compound represented by formula (1) is Fmoc-Lys·HCl, and the compound represented by formula (II) is Mtt-Cl. The present application has no special restrictions on their sources, and they can be purchased on the market.
[0035] In a specific implementation, the present application first mixes the compound represented by formula (1) with an organic solvent, then adds the compound represented by formula (II) and mixes them evenly, and then adds an organic base to react to obtain Fmoc-Lys(Mtt)-OH represented by formula (III).
[0036] The present application first mixes the compound shown in formula (I) with an organic solvent under stirring. The present application has no particular restrictions on the parameters of the stirring, and the compound of formula (I) and the organic solvent are uniformly mixed. In some specific implementations, the organic solvent of the reaction is selected from dichloromethane, methanol, ethanol or tetrahydrofuran. The present application has no particular restrictions on the amount of the organic solvent, as long as it is conducive to the reaction.
[0037] After the compound represented by formula (I) is mixed with an organic solvent, the compound represented by formula (II) is added under stirring and mixed. In some specific implementations, the compound represented by formula (II) is added at a reaction temperature, for example, 5°C to 25°C.
[0038] After adding the compound shown in formula (II) and mixing evenly, an organic base is added to react, preferably by adding the organic base dropwise. In some specific implementations, the organic base is selected from at least one of triethylamine or N,N-diisopropylethylamine, preferably N,N-diisopropylethylamine. In some specific implementations, the reaction temperature is 5°C to 25°C, preferably 15°C to 25°C, and the reaction time is 2h to 10h, preferably 2h to 6h. In the present application, the reaction temperature is lower than 5°C and the reaction is not thorough, and the reaction impurities increase when the reaction temperature is higher than 25°C, resulting in a low yield.
[0039] In some specific implementations, the molar ratio of the compound represented by formula (1), the compound represented by formula (II) and the organic base is 1:(0.8-1.3):(2-2.5), preferably 1:1.05:2.5.
[0040] After the reaction is completed, the obtained reaction product is post-processed. The present application has no particular restrictions on the specific manner of the post-processing, which may include the following steps:
[0041] The obtained reaction product is washed with citric acid solution and brine in turn, dried, filtered and then crystallized to obtain Fmoc-Lys(Mtt)-OH represented by formula (III).
[0042] After obtaining the reaction product, it is first washed with a citric acid aqueous solution, the mass concentration of which is 5% to 15%, preferably 10%, and the washing times are preferably twice. After washing with the citric acid aqueous solution, it is washed again with saturated brine, and the washing times are preferably 3 times. After washing, the obtained organic phase is dried, for example, by drying with anhydrous sodium sulfate, filtered, and then concentrated.
[0043] The concentrated reaction product is crystallized, and the crystallization solvent includes but is not limited to petroleum ether. Specifically, the concentrated reaction product is cooled to 20°C to 25°C, and then a solvent is added, and the temperature is cooled to 0°C to 5°C for crystallization, and the final product is obtained after filtering and drying. In some specific implementations, it is preferably dried at 50°C to 55°C.
[0044] The present application uses Fmoc-Lys·HCl shown in formula (I) as a starting material, and reacts with Mtt-Cl shown in formula (II) in the presence of an organic base to synthesize Fmoc-Lys(Mtt)-OH shown in formula II(I), which simplifies the process, is simple to operate, has mild reaction conditions, reduces the difficulty of industrial production, does not involve precious raw materials, is conducive to controlling production costs, and obtains Fmoc-Lys(Mtt)-OH with high purity and high yield. Experimental results show that the yield of Fmoc-Lys(Mtt)-OH prepared by the preparation method provided in the present application is more than 80%, and the purity is more than 99%.
[0045] The preparation method of Fmoc-Lys(Mtt)-OH provided in the present application is described in detail below with reference to the examples.
[0046] Example 1
[0047] 405 g of dichloromethane was added to a 2L three-necked flask, and 40.5 g of Fmoc-Lys·HCl was added under stirring. The temperature was controlled at 15°C to 25°C, 30.8 g of Mtt-Cl was added, and 27.1 g of N,N-diisopropylethylamine was added dropwise. After the addition was completed, the temperature was controlled at 15°C to 25°C, and the reaction was carried out for 4 hours. TLC showed that Fmoc-Lys·HCl was completely reacted; the reaction solution was washed twice with 10% citric acid aqueous solution (80 g / time), the organic phase was washed three times with saturated brine (80 g / time), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to a residual 130 g in the system; the temperature was lowered to 20°C to 25°C, 100 g of petroleum ether was added, a large amount of solid was precipitated, the temperature was lowered to 0 to 5°C for crystallization for 2h to 4h, filtered, and dried at 50°C to 55°C to obtain 52.1 g of white solid with a yield of 83.4% and a purity of 99.77%.
[0048] The white solid was analyzed and the results are shown in Figure 1 and Figure 2 , Figure 1 The high performance liquid phase (HPLC) spectrum of the product prepared in Example 1, Figure 2 This is the infrared spectrum of the product prepared in Example 1. Figure 1 and Figure 2 It can be seen that the white solid prepared in this application is Fmoc-Lys(Mtt)-OH.
[0049] Example 2
[0050] Add 405 g of methanol to a 2L three-necked flask, add 40.5 g of Fmoc-Lys·HCl under stirring, control the temperature at 15℃~25℃, add 30.8 g of Mtt-Cl, add 27.1 g of N,N-diisopropylethylamine dropwise, control the temperature at 15℃~25℃, react for 5 hours, and TLC Fmoc-Lys·HCl was completely reacted, 80g of water was added to the reaction solution, the pH was adjusted to 7-7.5 with citric acid, the methanol in the system was basically concentrated to dryness, 240g of dichloromethane was added, the reaction solution was washed twice with 10% citric acid aqueous solution (80g / time), the organic phase was washed three times with saturated brine (80g / time), the organic phase was dried with anhydrous sodium sulfate, filtered, and concentrated to a residual 133g in the system; the temperature was lowered to 20°C-25°C, 100g of petroleum ether was added, a large amount of solid was precipitated, the temperature was lowered to 0°C-5°C for crystallization for 2-4 hours, filtered, and dried at 50°C-55°C to obtain 53.3g of white solid, with a yield of 85.3% and a purity of 99.82%.
[0051] The white solid was analyzed and the results are shown in Figure 3 and Figure 4 , Figure 3 The high performance liquid phase (HPLC) spectrum of the product prepared in Example 2, Figure 4This is the infrared spectrum of the product prepared in Example 2. Figure 3 and Figure 4 It can be seen that the white solid prepared in this application is Fmoc-Lys(Mtt)-OH.
[0052] Example 3
[0053] Add 1500 kg of dichloromethane to a 3000 L enamel reactor, add 150 kg of Fmoc-Lys·HCl under stirring, control the temperature at 15℃~25℃, add 114 kg of Mtt-Cl, add 119.5 kg of N,N-diisopropylethylamine dropwise, control the temperature at 15℃~25℃, react for 6 hours, and TLC Fmoc-Lys·HCl was completely reacted, the reaction solution was washed twice with 10% citric acid aqueous solution (300 kg / time), the organic phase was washed three times with saturated brine (300 kg / time), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to a residual amount of 300-400 L in the system; the temperature was lowered to 20°C-25°C, 300 kg of petroleum ether was added, a large amount of solid was precipitated, the temperature was lowered to 0°C-5°C for crystallization for 2-4 hours, filtered, and dried at 50°C-55°C to obtain 191.65 kg of white solid with a yield of 82.8% and a purity of 99.81%.
[0054] The white solid was analyzed and the results are shown in Figure 5 , Figure 6 and Figure 7 , Figure 5 The high performance liquid phase (HPLC) spectrum of the product prepared in Example 3, Figure 6 The infrared spectrum of the product prepared in Example 3, Figure 7 This is the NMR spectrum of the product prepared in Example 3. Figure 5 , Figure 6 and Figure 7 It can be seen that the white solid prepared in this application is Fmoc-Lys(Mtt)-OH.
[0055] Embodiment 4-5
[0056] This example investigates the effect of organic solvents on the product. Specifically, referring to the preparation method provided in Example 1, only the type of organic solvent was changed, and the other preparation steps and process parameters were the same as in Example 1. The product yield is shown in Table 1 below.
[0057] Table 1 Yield and purity of the products prepared in Examples 1 to 4 of the present application
[0058] Organic solvents Yield purity Example 1 Dichloromethane 83.4％ 99.77％ Example 2 Methanol 85.3％ 99.82％ Example 4 Ethanol 83.6％ 99.13％ Example 5 Tetrahydrofuran 83.2％ 99.34％
[0059] Comparative Example 1
[0060] This comparative example investigates the effect of organic base on the product. Specifically, referring to the preparation method provided in Example 1, only the type of organic base is changed, and the other preparation steps and process parameters are the same as those in Example 1. After testing, the product yield is shown in Table 2 below.
[0061] Table 2 Yield and purity of the products prepared in the examples and comparative examples of the present application
[0062] Organic base Yield purity Example 1 N,N-Diisopropylethylamine 83.4％ 99.8％ Comparative Example 1 Triethylamine 72.8％ 98.34％
[0063] Example 6, Comparative Examples 2 to 4
[0064] This example and comparative example investigate the effect of reaction temperature on the product. Specifically, referring to the preparation method provided in Example 1, only the reaction temperature is changed, and the other preparation steps and process parameters are the same as those in Example 1. After testing, the product yield and reaction time are shown in Table 3 below.
[0065] Table 3 Yield and purity of the products prepared in Examples 6 to 7 and Comparative Examples 2 to 3 of the present application
[0066]
[0067]
[0068] In summary, the present invention provides a method for preparing Fmoc-Lys(Mtt)-OH, which is suitable for scale-up. The synthesis method requires simple equipment, has good reaction efficiency, low production cost, and the purity of the final product is greater than 99.0%, and the yield is above 80%.
[0069] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A method for preparing Fmoc-Lys(Mtt)-OH, comprising the following steps: The compound represented by formula (1) reacts with the compound represented by formula (II) under the action of an organic base to obtain Fmoc-Lys(Mtt)-OH represented by formula (III); 2. The preparation method according to claim 1, characterized in that: The organic base is selected from at least one of triethylamine and N,N-diisopropylethylamine.
3. The preparation method according to claim 1, characterized in that: The molar ratio of the compound represented by formula (1), the compound represented by formula (II) and the organic base is 1:(0.8-1.3):(2-2.5).
4. The preparation method according to claim 1, characterized in that: The organic solvent for the reaction is selected from dichloromethane, methanol, ethanol or tetrahydrofuran.
5. The preparation method according to claim 1, characterized in that: The reaction temperature is 5°C to 25°C, and the reaction time is 2h to 10h.
6. The preparation method according to any one of claims 1 to 5, characterized in that: The compound represented by formula (1) is mixed evenly with an organic solvent, and then the compound represented by formula (II) is added and mixed evenly, and then an organic base is added to react.
7. The preparation method according to claim 6, characterized in that: The temperature for adding the compound represented by formula (II) is 5°C to 25°C.
8. The preparation method according to claim 6, characterized in that: The organic base is added dropwise.
9. The preparation method according to claim 8, characterized in that: The method also includes post-processing the obtained reaction product.
10. The preparation method according to claim 9, characterized in that: The post-processing includes: The obtained reaction product is washed with citric acid solution and brine in turn, dried, filtered and then crystallized to obtain Fmoc-Lys(Mtt)-OH represented by formula (III).