Preparation method of ifosfamide, intermediate and ifosfamide composition

Abstract

The invention provides a preparation method of ifosfamide, an intermediate and an ifosfamide composition, and belongs to the technical field of medicines. Comprising the following steps: S1, carrying out cyclization reaction on phosphorus oxychloride and n-propanolamine under the action of an acid-binding agent to obtain a compound 1; s2, the compound 1 and 3-halo-propane-1-alcohol are subjected to a reaction, and a compound 2 is prepared; s3, reacting the compound 2 with ethanolamine to prepare a compound 3; and S4, carrying out a reaction on the compound 3 and thionyl chloride to prepare the ifosfamide. The preparation method of the ifosfamide is simple, mild in reaction condition, obviously improved in total yield, fewer in by-products and higher in product purity, and has a wide application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of medicine, and in particular to a preparation method of ifosfamide, an intermediate and an ifosfamide composition. Background Art

[0002] Ifosfamide, also known as Helexan, is an isomer of cyclophosphamide. It was first developed and synthesized by the German company Asta in the mid-1960s, but it did not enter clinical use until the 1980s, with the advent of the urinary tract protectant Mesna. It is now widely used worldwide. Ifosfamide is a broad-spectrum anti-tumor agent and a second-generation alkylating agent with inhibitory effects on various tumors. Its mechanism of action is to cross-link with DNA, inhibiting DNA synthesis. It can also interfere with RNA function, making it a non-specific agent for the cell cycle.

[0003] Currently, the synthesis methods of ifosfamide mainly include the following methods: Route 1: .

[0004] This route uses ethyleneimine as the starting material and obtains ifosfamide through a three-step reaction. The reaction process is relatively complicated and the yield is only about 20%. At the same time, the ethyleneimine used is highly toxic and requires a large amount. In the first step of the reaction, the amount of ethyleneimine required is 10 times that of 3-chloropropanol. Although it can be subsequently recovered by distillation, the recovery is not complete and there is a large amount of residue, making it unsuitable for industrial production.

[0005] Route 2: .

[0006] This route uses phosphorus oxychloride as the starting material and obtains ifosfamide through a four-step reaction, with the total yield increased to about 30%. However, in the second step, side reactions are prone to occur during the reaction between chloroethylamine hydrochloride and 2-chlorotetrahydro-2H-1,3,2-oxazinophosphorus-2-oxide, affecting the final yield.

[0007] Chinese invention patent CN115505003B discloses a method for preparing ifosfamide and a method for preparing ifosfamide for injection, the route used is as follows: .

[0008] Although the total yield in this reaction is increased to more than 36%, in the third step, chloroethylamine hydrochloride easily undergoes a nucleophilic substitution reaction with an alkyl chloride, thereby increasing the amount of by-products and reducing the reaction yield.

[0009] Chinese invention patent CN111943979B discloses an ifosfamide intermediate and its preparation method and application. The synthesis route is as follows: .

[0010] This reaction proposed a new synthesis method for ifosfamide, with an overall yield of 39.8%. However, in the third step, 3-bromo-propan-1-ol reacts not only with secondary amines but also with alcoholic hydroxyl groups, resulting in an increase in reaction byproducts and a significant decrease in the overall yield. Summary of the Invention

[0011] The present invention aims to provide a preparation method, intermediate and ifosfamide composition of ifosfamide. The preparation method is simple, the total yield is significantly improved, the by-products are relatively few, and the product purity is high. A hydrogen bond structure is formed between ifosfamide and a porous sugar microsphere scaffold, which inhibits intermolecular agglomeration, thereby significantly improving the glass transition temperature thereof. The group capable of forming hydrogen bonds with water molecules is reduced, thereby reducing hygroscopicity, overcoming the industry problems of ifosfamide hygroscopicity and sintering, and having broad application prospects.

[0012] The technical solution of the present invention is achieved as follows: The present invention provides a method for preparing ifosfamide, which comprises the following steps: S1. Phosphorus oxychloride and n-propanolamine were subjected to a cyclization reaction in the presence of an acid binding agent to give compound 1; S2. Compound 1 is reacted with 3-halogen-propane-1-ol to obtain compound 2; S3. Compound 2 is reacted with ethanolamine to obtain compound 3; S4. Compound 3 is reacted with thionyl chloride to obtain ifosfamide; The specific chemical reaction formula is as follows: As a further improvement of the present invention, the 3-halogen-propane-1-ol in step S2 is selected from at least one of 3-bromo-propane-1-ol and 3-chloro-propane-1-ol, the molar ratio of compound 1 to 3-halogen-propane-1-ol is 1-1.1:1, triethylamine is also added to the reaction system, and the reaction conditions are stirring at room temperature for 2-4 hours.

[0013] As a further improvement of the present invention, the molar ratio of compound 2 to ethanolamine in step S3 is 1:1.1-1.2, triethylamine is also added to the reaction system, and the reaction conditions are 30-40° C. and stirring for 2-4 hours.

[0014] As a further improvement of the present invention, the molar ratio of compound 3 to thionyl chloride in step S4 is 1:2.5-3, and the reaction conditions are stirring at room temperature for 1-3 hours.

[0015] The present invention further protects an ifosfamide synthesis intermediate, the structural formula of which is shown in Formula I: Formula I.

[0016] The present invention further protects a method for preparing the above-mentioned ifosfamide synthetic intermediate, which is prepared by reacting a compound represented by Formula II with ethanolamine: Formula II.

[0017] The present invention further protects a method for preparing an ifosfamide composition, comprising the following steps: S1. Dissolve sodium alginate and glucose in water, add a porogen and an emulsifier, and stir to form an aqueous phase; S2. The aqueous phase was added to the fish oil, emulsified, and a calcium chloride solution was added dropwise. The mixture was solidified at room temperature, centrifuged, washed, and dried to prepare a porous sugar microsphere scaffold. S3. Dissolve ifosfamide in water, add the porous sugar microsphere scaffold, stir and mix evenly, and freeze-dry to prepare an ifosfamide composition.

[0018] As a further improvement of the present invention, the mass ratio of sodium alginate, glucose, porogen and emulsifier in step S1 is 10-12:3-5:1-2:2-3, the porogen is selected from at least one of cetyltrimethylammonium chloride, cetyltrimethylammonium bromide, cetyltrimethyl sodium chloride and cetyltrimethyl sodium bromide, and the emulsifier is selected from at least one of Tween-20, Tween-40, Tween-60 and Tween-80.

[0019] As a further improvement of the present invention, the mass ratio of ifosfamide to the porous sugar microsphere scaffold in step S3 is 8-10:3-5.

[0020] The present invention further protects an ifosfamide composition prepared by the above preparation method.

[0021] The present invention has the following beneficial effects: The synthetic route of the present invention is as follows: .

[0022] The present invention prepares compound 2 by reacting with 3-halogen-propane-1-ol on the basis of the existing compound 1. This process produces fewer by-products. Compound 2 is further reacted with ethanolamine. Since the substitution reaction activity of amino group with chlorine is significantly stronger than that of alcoholic hydroxyl group, the yield of this step is higher and the by-products are fewer. A product mainly composed of compound 3 can be obtained. Subsequently, ifosfamide is prepared by unified chlorination with thionyl chloride. Through the reaction of the present invention, the total yield is significantly improved, the by-products are fewer, and the product purity is higher.

[0023] In addition, the present invention prepares an ifosfamide composition, by first preparing a porous sugar microsphere scaffold, adsorbing ifosfamide and then freeze-drying, forming a hydrogen bond structure between ifosfamide and the porous sugar microsphere scaffold, inhibiting intermolecular agglomeration, thereby significantly improving its glass transition temperature, and reducing the groups that can form hydrogen bonds with water molecules, thereby reducing hygroscopicity, overcoming the industry problems of ifosfamide hygroscopicity and sintering, and having broad application prospects. DETAILED DESCRIPTION

[0024] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0025] Example 1 This embodiment provides a method for preparing ifosfamide, comprising the following steps: S1. Add 0.1 mol of phosphorus oxychloride to 100 mL of dichloromethane. At 4°C, add dropwise 50 mL of a dichloromethane solution containing 0.1 mol of n-propanolamine and 0.2 mol of triethylamine. Warm to room temperature and stir for 3 h to allow the cyclization reaction to proceed. Filter and concentrate the filtrate under reduced pressure. Add 100 mL of tetrahydrofuran, filter, and concentrate under reduced pressure. Separate and purify by column chromatography to obtain compound 1 with a yield of 72.5%. ESI-MS calculated value: C3H8ClNO2P(M+H) + 155.99, measured value: 156.0.

[0026] NMR results: 1 H NMR (300MHz, CDCl3) δ4.10 (t, 2H), 2.58 (t, 2H), 1.95-2.0 (m, 3H); S2. 0.1 mol of compound 1, 0.1 mol of 3-bromo-propan-1-ol, and 0.15 mol of triethylamine were added to 150 mL of dichloromethane and stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure and purified by column chromatography to obtain compound 2 in an 83.2% yield. ESI-MS calculated value: C5H 12 ClNO3P(M+H) + 200.02, measured value: 200.0.

[0027] NMR results: 1 H NMR (300MHz, CDCl3) δ4.04 (t, 2H), 3.59 (t, 2H), 2.67 (t, 2H), 2.47 (t, 2H), 2.0 (br, 1H), 1.67 (m, 2H); S3. 0.1 mol of compound 2, 0.11 mol of ethanolamine, and 0.12 mol of triethylamine were reacted, added to 120 mL of dichloromethane, and stirred at room temperature for 2 h. The reaction was concentrated under reduced pressure and purified by column chromatography to obtain compound 3 in a yield of 78.9%. ESI-MS calculated value: C8H 18 N2O4P(M+H) + 225.09, measured value: 225.1.

[0028] NMR results: 1 H NMR (300MHz, CDCl3) δ4.01 (t, 2H), 3.85 (t, 2H), 3.72 (t, 2H), 2.78 (t, 2H), 2.69 (t, 2H), 2.43 (t, 2H), 2.0-2.2 (br, 3H), 1.71 (m, 2H); S4. 0.1 mol of compound 3 was added to 100 mL of dichloromethane, followed by the addition of 50 mL of a dichloromethane solution containing 0.2 mol of thionyl chloride. The reaction was stirred at room temperature for 1 h. The solvent and excess thionyl chloride were removed under reduced pressure, and the mixture was washed and dried to obtain ifosfamide. The yield was 94.2% and the purity was 99.94%. ESI-MS calculated value: C7H 16 Cl2N2O2P(M+H) + 261.02, found value: 261.0, melting point 48-51℃, NMR results are consistent with the literature.

[0029] Example 2 This embodiment provides a method for preparing ifosfamide, comprising the following steps: S1. 0.1 mol of phosphorus oxychloride was added to 100 mL of dichloromethane. 50 mL of a dichloromethane solution containing 0.11 mol of n-propanolamine and 0.25 mol of triethylamine was added dropwise at 4°C. The mixture was allowed to warm to room temperature and stirred for 3 hours for a cyclization reaction. The mixture was filtered and concentrated under reduced pressure. 100 mL of tetrahydrofuran was added, the mixture was filtered, and the mixture was concentrated under reduced pressure. Compound 1 was isolated and purified by column chromatography to obtain compound 1 in a yield of 72.9%. S2. 0.11 mol of compound 1, 0.1 mol of 3-bromo-propan-1-ol, and 0.15 mol of triethylamine were added to 150 mL of dichloromethane. The reaction was stirred at room temperature for 4 h. The mixture was concentrated under reduced pressure and purified by column chromatography to obtain compound 2 in a yield of 84.5%. S3. 0.1 mol of compound 2, 0.12 mol of ethanolamine, and 0.12 mol of triethylamine were reacted in 120 mL of dichloromethane. The reaction was stirred at room temperature for 4 h, concentrated under reduced pressure, and purified by column chromatography to obtain compound 3 in a yield of 79.5%. S4. Add 0.1 mol of compound 3 to 100 mL of dichloromethane, followed by 50 mL of a dichloromethane solution containing 0.3 mol of thionyl chloride. Stir the reaction at room temperature for 3 h. Remove the solvent and excess thionyl chloride under reduced pressure, wash, and dry to obtain ifosfamide in a 95.3% yield and 99.95% purity.

[0030] Example 3 This embodiment provides a method for preparing ifosfamide, comprising the following steps: S1. Add 0.1 mol of phosphorus oxychloride to 100 mL of dichloromethane. At 4°C, add dropwise 50 mL of a dichloromethane solution containing 0.1 mol of n-propanolamine and 0.2 mol of triethylamine. Warm to room temperature and allow the cyclization reaction to proceed with stirring for 3 hours. Filter and concentrate the filtrate under reduced pressure. Add 100 mL of tetrahydrofuran, filter, and concentrate under reduced pressure. Separate and purify by column chromatography to obtain compound 1 in a yield of 72.5%. S2. 0.105 mol of compound 1, 0.1 mol of 3-chloro-propane-1-ol, and 0.15 mol of triethylamine were added to 150 mL of dichloromethane. The reaction was stirred at room temperature for 3 h. The mixture was concentrated under reduced pressure and purified by column chromatography to obtain compound 2 in a yield of 83.9%. S3. 0.1 mol of compound 2, 0.115 mol of ethanolamine, and 0.12 mol of triethylamine were reacted in 120 mL of dichloromethane. The reaction was stirred at room temperature for 3 h, concentrated under reduced pressure, and purified by column chromatography to obtain compound 3 in a yield of 79.2%. S4. Add 0.1 mol of compound 3 to 100 mL of dichloromethane, followed by 50 mL of a dichloromethane solution containing 0.25 mol of thionyl chloride. Stir the reaction at room temperature for 2 h. Remove the solvent and excess thionyl chloride under reduced pressure, wash, and dry to obtain ifosfamide in a yield of 94.8% and a purity of 99.97%.

[0031] Example 4 This embodiment provides a method for preparing an ifosfamide composition, comprising the following steps: S1. Dissolve 10g of sodium alginate and 3g of glucose in 200mL of water, add 1g of hexadecyltrimethylammonium bromide and 2g of Tween-40, and stir to form an aqueous phase; S2. The aqueous phase was added to 500 mL of fish oil and emulsified at 8000 rpm for 15 min. 50 mL of a 2 wt% calcium chloride solution was added dropwise and the mixture was cured at room temperature for 30 min. The mixture was centrifuged, washed, and dried to obtain a porous sugar microsphere scaffold. S3. Dissolve 8 g of ifosfamide in 200 mL of water, add 3 g of the porous sugar microsphere scaffold, stir and mix for 20 minutes, and freeze-dry to prepare an ifosfamide composition.

[0032] Example 5 This embodiment provides a method for preparing an ifosfamide composition, comprising the following steps: S1. Dissolve 12g of sodium alginate and 5g of glucose in 200mL of water, add 2g of hexadecyltrimethylammonium bromide and 3g of Tween-60, and stir to form an aqueous phase; S2. The aqueous phase was added to 500 mL of fish oil and emulsified at 8000 rpm for 15 min. 50 mL of a 2 wt% calcium chloride solution was added dropwise and the mixture was cured at room temperature for 30 min. The mixture was centrifuged, washed, and dried to obtain a porous sugar microsphere scaffold. S3. Dissolve 10 g of ifosfamide in 200 mL of water, add 5 g of the porous sugar microsphere scaffold, stir and mix for 20 minutes, and freeze-dry to prepare an ifosfamide composition.

[0033] Example 6 This embodiment provides a method for preparing an ifosfamide composition, comprising the following steps: S1. Dissolve 11g of sodium alginate and 4g of glucose in 200mL of water, add 1.5g of hexadecyltrimethylammonium chloride and 2.5g of Tween-80, and stir to form an aqueous phase; S2. The aqueous phase was added to 500 mL of fish oil and emulsified at 8000 rpm for 15 min. 50 mL of a 2 wt% calcium chloride solution was added dropwise and the mixture was cured at room temperature for 30 min. The mixture was centrifuged, washed, and dried to obtain a porous sugar microsphere scaffold. S3. Dissolve 9 g of ifosfamide in 200 mL of water, add 4 g of the porous sugar microsphere scaffold, stir and mix for 20 minutes, and freeze-dry to prepare an ifosfamide composition.

[0034] Comparative Example 1 Compared with Example 3, the difference is that no porous sugar microsphere scaffold is prepared, and glucose and ifosfamide are directly mixed and freeze-dried.

[0035] 9 g of ifosfamide was dissolved in 200 mL of water, 4 g of glucose was added, the mixture was stirred for 20 minutes, and freeze-dried to prepare an ifosfamide composition.

[0036] Test Example 1 The ifosfamide compositions prepared in Examples 4-6 and Comparative Example 1 and commercially available ifosfamide were stored at a temperature of 40° C. and a humidity of 85 RH% for 6-9 months. The observation results are shown in Table 1.

[0037] Table 1 As can be seen from the table above, the ifosfamide compositions prepared in Examples 4-6 of the present invention overcome the industry problem of ifosfamide sintering and are not easily sintered. Comparative Example 1 significantly alleviates sintering compared to commercially available products; however, sintering also occurs after storage for more than 7 months. Therefore, the ifosfamide compositions prepared in Examples 4-6 of the present invention have better effects.

[0038] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing ifosfamide, characterized in that: The preparation method comprises the following steps: S1. Phosphorus oxychloride and n-propanolamine were subjected to a cyclization reaction in the presence of an acid binding agent to give compound 1; S2. Compound 1 is reacted with 3-halogen-propane-1-ol to obtain compound 2; S3. Compound 2 is reacted with ethanolamine to obtain compound 3; S4. Compound 3 is reacted with thionyl chloride to obtain ifosfamide; The specific chemical reaction formula is as follows: 。 2. The preparation method according to claim 1, characterized in that The 3-halogen-propane-1-ol in step S2 is selected from at least one of 3-bromo-propane-1-ol and 3-chloro-propane-1-ol, the molar ratio of compound 1 to 3-halogen-propane-1-ol is 1-1.1:1, triethylamine is also added to the reaction system, and the reaction conditions are stirring at room temperature for 2-4 hours.

3. The preparation method according to claim 1, characterized in that In step S3, the molar ratio of compound 2 to ethanolamine is 1:1.1-1.2, triethylamine is also added to the reaction system, and the reaction conditions are 30-40° C. and stirring for 2-4 hours.

4. The preparation method according to claim 1, characterized in that In step S4, the molar ratio of compound 3 to thionyl chloride is 1:2.5-3, and the reaction is stirred at room temperature for 1-3 hours.

5. A synthetic intermediate of ifosfamide, characterized in that: Its structural formula is shown in Formula I: Formula I.

6. The method for preparing the ifosfamide synthetic intermediate according to claim 5, characterized in that: The compound represented by formula II is reacted with ethanolamine to prepare: Formula II.

7. A method for preparing an ifosfamide composition, characterized in that: The following steps are involved: S1. Dissolve sodium alginate and glucose in water, add a porogen and an emulsifier, and stir to form an aqueous phase; S2. The aqueous phase was added to the fish oil, emulsified, and a calcium chloride solution was added dropwise. The mixture was solidified at room temperature, centrifuged, washed, and dried to prepare a porous sugar microsphere scaffold. S3. Dissolve ifosfamide in water, add the porous sugar microsphere scaffold, stir and mix evenly, and freeze-dry to prepare an ifosfamide composition.

8. The preparation method according to claim 7, characterized in that The mass ratio of sodium alginate, glucose, porogen and emulsifier in step S1 is 10-12:3-5:1-2:2-3, the porogen is selected from at least one of cetyltrimethylammonium chloride, cetyltrimethylammonium bromide, cetyltrimethyl sodium chloride and cetyltrimethyl sodium bromide, and the emulsifier is selected from at least one of Tween-20, Tween-40, Tween-60 and Tween-80.

9. The preparation method according to claim 7, characterized in that The mass ratio of ifosfamide to the porous sugar microsphere scaffold in step S3 is 8-10:3-5.

10. An ifosfamide composition obtained by the preparation method according to any one of claims 7 to 9.

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