Gelated and / or micronized nucleoside medicine and preparation method thereof

Abstract

The invention discloses a gelatinized and / or micronized nucleoside drug and a preparation method thereof, and belongs to the technical field of drugs. The preparation method comprises the following steps: mixing lipoic acid, a nucleoside drug and an esterification catalyst, carrying out a grafting reaction in a reaction solvent to obtain a lipoic acid grafted nucleoside drug, dispersing the lipoic acid grafted nucleoside drug in an organic solvent to obtain a dispersion liquid, dropwise adding the dispersion liquid into hot water, and carrying out ring opening polymerization to obtain the lipoic acid grafted nucleoside drug. The nucleoside drug nanogel, nanoparticle or nanofiber is obtained, and the structure and performance improvement based on the drug itself is realized. The gelated and / or micronized nucleoside medicine prepared by the invention has obviously improved scavenging rates of DPPH free radicals and ABTS free radicals, and the method can improve the oxidation resistance of the nucleoside medicine and has the characteristic of improving the stability and functionality of the medicine.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a gelled and / or microparticulated nucleoside drug and a preparation method thereof. BACKGROUND

[0002] Nucleoside drugs interfere with the synthesis of viral nucleic acids by mimicking the structure of natural nucleosides. Their antiviral activity mainly depends on the following core processes: selective phosphorylation, competitive inhibition with natural substrates, and chain termination reactions. Anti-herpes virus drugs are one of the most successful applications of nucleoside drugs, with representative drugs including acyclovir and ganciclovir. These drugs have high selectivity, rely on viral-encoded kinases for activation, significantly reducing toxicity to host cells, and are widely used in clinical practice and in-depth research by inhibiting viral DNA replication and assisting the host immune system in clearing viruses. However, drugs such as acyclovir and ganciclovir still face problems such as drug resistance, suboptimal pharmacokinetic properties, and functional deficiencies, necessitating improvements in their stability and functionality.

[0003] Currently, the main methods for improving the efficacy of nucleoside drugs include drug delivery systems using gels, DNA tetrahedrons, nanomaterials, and other carriers, combining photothermal dynamic therapy, or using penetration-enhancing drugs to improve drug permeability in the human body. These methods often introduce new problems, such as the lack of multifunctional properties in the drugs themselves, such as antioxidant properties, the use of carriers to load drugs often faces problems such as uncontrollable drug dispersion and release rate, and the combination of photothermal dynamics often requires the introduction of photosensitizers and light sources. While penetration-enhancing drugs can improve the permeability of nucleoside drugs to some extent, they cannot avoid the harm to the human body caused by penetration-enhancing drugs. SUMMARY

[0004] In view of the above prior art, the present application provides a gelled and / or microparticulated nucleoside drug and a preparation method thereof to solve the technical problems of suboptimal pharmacokinetic properties and insufficient functionality of nucleoside drugs.

[0005] To achieve the above-mentioned purpose, the technical solution adopted by the present application is to provide a preparation method of a gelled and / or microparticulated nucleoside drug, comprising the following steps: S1: grafting lipoic acid to a nucleoside drug to obtain a nucleoside drug grafted with lipoic acid; S2: ring-opening polymerization of the nucleoside drug grafted with lipoic acid to obtain a gelled and / or microparticulated nucleoside drug.

[0006] On the basis of the above technical solution, the present application can also be improved as follows.

[0007] Further, the method for grafting lipoic acid with nucleoside drugs in S1 is: The lipoic acid, nucleoside drugs and esterification catalyst are co-dissolved in a reaction solvent, and stirred at room temperature for 12-24 h to obtain the product; the esterification catalyst is a mixture of 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4-dimethylaminopyridine.

[0008] Further, the nucleoside drugs are acyclovir, the reaction solvent is dimethyl sulfoxide, and the esterification catalyst is obtained by mixing 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4-dimethylaminopyridine at a molar ratio of 1:1; the molar ratio of lipoic acid, acyclovir and esterification catalyst is 1:1:3; and the material-liquid ratio of acyclovir to dimethyl sulfoxide is 1 mol:20 L.

[0009] Further, the reaction time for grafting lipoic acid with acyclovir is 12 h.

[0010] Further, the nucleoside drugs are ganciclovir, the reaction solvent is dimethyl sulfoxide, and the esterification catalyst is obtained by mixing 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4-dimethylaminopyridine at a molar ratio of 1:1; the molar ratio of lipoic acid, ganciclovir and esterification catalyst is 1:2:6; and the material-liquid ratio of ganciclovir to dimethyl sulfoxide is 1 mol:10 L.

[0011] Further, the reaction time for grafting lipoic acid with ganciclovir is 24 h.

[0012] Further, the method for ring-opening polymerization in S2 is: The nucleoside drugs grafted with lipoic acid are dispersed in an organic solvent to obtain a dispersion liquid with a concentration of 1-5 mg / mL; then the dispersion liquid is dropped into water at 80-100 DEG C, and stirred and polymerized for 2-24 h to obtain the product.

[0013] Further, the prepared drug is a gelled nucleoside drug; the concentration of the dispersion liquid is 5 mg / mL; and the stirring and polymerization time is 12 h.

[0014] Further, the prepared drug is a micronized nucleoside drug; the concentration of the dispersion liquid is 1 mg / mL; and the stirring and polymerization time is 2 h.

[0015] The application further discloses the gelled and / or micronized nucleoside drugs prepared by the preparation method; wherein the micronized nucleoside drugs are nucleoside drug nanoparticles or nucleoside drug nanofibers; the particle size of the nucleoside drug nanoparticles is 200-350 nm; and the width of the nucleoside drug nanofibers is 200-400 nm, and the length-width ratio is 1.5-3.

[0016] The application has the following beneficial effects: 1. The present application induces ring-opening polymerization by grafting lipoic acid on nucleoside drugs to obtain drug gels or drug microparticles, which have significantly improved DPPH free radical and ABTS free radical scavenging rates, and the method of the present application can improve the antioxidant properties of nucleoside drugs, and has the characteristics of improving drug stability and functionality.

[0017] 2. The present application mixes lipoic acid, nucleoside drugs and esterification catalysts, carries out grafting reaction in a reaction solvent to obtain lipoic acid grafted nucleoside drugs, disperses the lipoic acid grafted nucleoside drugs in an organic solvent to obtain a dispersion liquid, drops the dispersion liquid into hot water to induce ring-opening polymerization, and obtains nucleoside drug nanogels, nanoparticles or nanofibers, thereby realizing improvement based on the structure and performance of the drugs themselves. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 The reaction equation for grafting lipoic acid on acyclovir is as follows: Figure 2 The reaction equation for grafting lipoic acid on ganciclovir is as follows: Figure 3 The nuclear magnetic resonance hydrogen spectrum of acyclovir grafted with lipoic acid is as follows: Figure 4 The mass spectrum of acyclovir grafted with lipoic acid is as follows: Figure 5 The scanning electron microscope image of acyclovir nanogels obtained in Example 1 is as follows: Figure 6 The mass spectrum of acyclovir nanogels obtained in Example 1 is as follows: Figure 7 The scanning electron microscope image of acyclovir nanoparticles obtained in Example 2 is as follows: Figure 8 The nuclear magnetic resonance hydrogen spectrum of ganciclovir grafted with lipoic acid is as follows: Figure 9 The mass spectrum of ganciclovir grafted with lipoic acid is as follows: Figure 10 The scanning electron microscope image of ganciclovir nanogels obtained in Example 3 is as follows: Figure 11 The scanning electron microscope image of ganciclovir nanofibers obtained in Example 4 is as follows: Figure 12 The antioxidant performance test results of acyclovir grafted with lipoic acid obtained in Example 1 are as follows: Figure 13 The rheological test results of acyclovir nanogels obtained in Example 1 are as follows: Figure 14 The antioxidant performance test results of acyclovir nanogels obtained in Example 1 are as follows. DETAILED DESCRIPTION

[0019] The technical principle of the technical scheme of the present application is that lipoic acid is used as a grafting group to combine with the hydroxyl group in nucleoside drugs to modify the nucleoside drugs, and lipoic acid grafted nucleoside drugs are obtained; then ring-opening polymerization of the lipoic acid grafted nucleoside drugs is induced to prepare gelled and / or microparticulated nucleoside drugs. The nucleoside drugs used in the present application include acyclovir and ganciclovir; wherein the structural formula of acyclovir is shown as formula I, and the structural formula of ganciclovir is shown as formula II:

[0020] The lipoic acid used has a carboxyl group at one end and a dithiolane ring at the other end; the carboxyl group of lipoic acid is coupled with the hydroxyl group of the nucleoside drug molecule through esterification reaction to obtain nucleoside drugs with polymerization ability and antioxidant ability.

[0021] The reaction equations of acyclovir and ganciclovir grafting lipoic acid are shown as Figure 1 and Figure 2 The process for grafting lipoic acid to nucleoside drug molecules is as follows: Lipoic acid, nucleoside drugs and esterification catalysts are mixed to perform grafting reaction in a reaction solvent to obtain lipoic acid grafted nucleoside drugs. The molar ratio of lipoic acid, nucleoside drugs and esterification catalysts is 1: (1-2): (3-6), the grafting reaction is performed under stirring, and the time is 12-24 h; the esterification catalyst is 1-ethyl- (3-dimethylaminopropyl) carbonyldiimide and 4-dimethylaminopyridine; the reaction solvent is dimethyl sulfoxide or dimethylformamide.

[0022] The structural formulas of lipoic acid grafted acyclovir and lipoic acid grafted ganciclovir are shown as formula III and formula IV, respectively:

[0023] The ring-opening polymerization process of lipoic acid grafted nucleoside drugs in the present application is as follows: The lipoic acid grafted nucleoside drugs are dispersed in an organic solvent to obtain a dispersion liquid, the dispersion liquid is dropped into hot water at 80-100 ℃, and stirring polymerization is performed for 2-24 h; the concentration of lipoic acid grafted nucleoside drugs in the dispersion liquid is 1-5 mg / mL; the organic solvent is dimethyl sulfoxide or dimethylformamide; specifically, if nucleoside drug nanogel is to be obtained, the stirring polymerization time is 12 h, and the concentration of lipoic acid grafted nucleoside drugs in the dispersion liquid is 5 mg / mL; if microparticulated nucleoside drugs are to be obtained, the stirring polymerization time is 2 h, and the concentration of lipoic acid grafted nucleoside drugs in the dispersion liquid is 1 mg / mL.

[0024] The nucleoside drug microparticles prepared by the preparation method of the application comprise nucleoside drug nanoparticles or nucleoside drug nanofibers; wherein the particle size of the nucleoside drug nanoparticles is 200-350 nm; the width of the nucleoside drug nanofibers is 200-400 nm, and the length-width ratio is 1.5-3.

[0025] The specific embodiments of the application will be described in detail below with reference to examples.

[0026] Example 1 An acyclovir nanogel is prepared by the following steps: (1) 1 mmol of lipoic acid, 3 mmol of esterification catalyst and 1 mmol of acyclovir are mixed and dissolved in 20 mL of dimethyl sulfoxide, and stirred at room temperature for 12 h to obtain acyclovir grafted with lipoic acid; wherein the esterification catalyst is 1.5 mmol of 1-ethyl-(3-dimethylaminopropyl) carbonyldiimide and 1.5 mmol of 4-dimethylamino pyridine; (2) The acyclovir grafted with lipoic acid is dispersed in dimethyl sulfoxide to obtain a dispersion liquid, the concentration of the acyclovir grafted with lipoic acid in the dispersion liquid is 5 mg / mL, the dispersion liquid is dropped into hot water at 90 ℃, and stirred at 90 ℃ for 12 h to make ring-opening polymerization, thereby obtaining the acyclovir nanogel.

[0027] Example 2 An acyclovir nanoparticle is prepared by the following steps: (1) 1 mmol of lipoic acid, 3 mmol of esterification catalyst and 1 mmol of acyclovir are mixed and dissolved in 20 mL of dimethyl sulfoxide, and stirred at room temperature for 12 h to obtain acyclovir grafted with lipoic acid; wherein the esterification catalyst is 1.5 mmol of 1-ethyl-(3-dimethylaminopropyl) carbonyldiimide and 1.5 mmol of 4-dimethylamino pyridine; (2) The acyclovir grafted with lipoic acid is dispersed in dimethyl sulfoxide to obtain a dispersion liquid, the concentration of the acyclovir grafted with lipoic acid in the dispersion liquid is 1 mg / mL, the dispersion liquid is dropped into hot water at 90 ℃, and stirred at 90 ℃ for 2 h to make ring-opening polymerization, thereby obtaining the acyclovir nanoparticle.

[0028] Example 3 A ganciclovir nanogel is prepared by the following steps: (1) 1 mmol lipoic acid, 6 mmol esterification catalyst and 2 mmol ganciclovir were mixed and dissolved in 20 mL dimethyl sulfoxide, and stirred at room temperature for 24 h to obtain ganciclovir grafted with lipoic acid; wherein the esterification catalyst was 3 mmol 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4 mmol 4-dimethylaminopyridine; (2) The ganciclovir grafted with lipoic acid was dispersed in dimethyl sulfoxide to obtain a dispersion liquid, and the concentration of ganciclovir grafted with lipoic acid in the dispersion liquid was 5 mg / mL; the dispersion liquid was dropped into hot water at 90°C, and stirred at 90°C for 12 h to make ring-opening polymerization, thereby obtaining ganciclovir nanogel.

[0029] Example 4 A ganciclovir nanofiber was prepared by the following steps: (1) 1 mmol lipoic acid, 6 mmol esterification catalyst and 2 mmol ganciclovir were mixed and dissolved in 20 mL dimethyl sulfoxide, and stirred at room temperature for 24 h to obtain ganciclovir grafted with lipoic acid; wherein the esterification catalyst was 3 mmol 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4 mmol 4-dimethylaminopyridine; (2) The ganciclovir grafted with lipoic acid was dispersed in dimethyl sulfoxide to obtain a dispersion liquid, and the concentration of ganciclovir grafted with lipoic acid in the dispersion liquid was 1 mg / mL; the dispersion liquid was dropped into hot water at 90°C, and stirred at 90°C for 2 h to make ring-opening polymerization, thereby obtaining ganciclovir nanofiber glue.

[0030] Experimental Example I. Morphological characterization of gelled and / or microparticulated nucleoside drugs The product of step (1) of Example 1 was subjected to nuclear magnetic resonance detection, and the hydrogen spectrum and mass spectrum are shown in Figure 3 and 4 , Figure 4 The characteristic peaks of the target product are in the boxes; as can be seen from Figure 3 and 4 , lipoic acid grafting is successful, and the molecular weight of the target product is 413.

[0031] The scanning electron microscope picture of the acyclovir nanogel of Example 1 is shown in Figure 5 , which shows a crosslinked network structure, and the presence of nanoparticles in the skeleton can be observed, which is a gel network formed by the mutual crosslinking of nanoparticles. The mass spectrum analysis result of the acyclovir nanogel is shown in Figure 6The box in the figure is the characteristic peak of the target product; it can be seen that the gel molecular weight of this embodiment is 826, which matches the molecular weight of the two lipoic acid grafted acyclovir, indicating that the ring-opening polymerization from a polymer to a dimer is achieved in this embodiment.

[0032] The acyclovir nanoparticles of Example 2 were subjected to scanning electron microscope test, and the results are shown in Figure 7 , wherein, Figure 7 (a) is a scanning electron microscope image of the nanoparticles, Figure 7 (b) is a particle size distribution graph of the nanoparticles; it can be seen from Figure 7 that the prepared acyclovir nanoparticles are mostly spherical with a particle size of 200-350 nm.

[0033] The product of Example 3 step (1) was subjected to nuclear magnetic resonance detection, and the hydrogen spectrum and mass spectrum are shown in Figure 8 and 9 , wherein, Figure 9 the box in the figure is the characteristic peak of the target product. It can be seen from Figure 8 and 9 that lipoic acid grafting is successful, and the molecular weight of the target product is 631.

[0034] The scanning electron microscope image of the ganciclovir nanogel of Example 3 is shown in Figure 10 . It can be seen from Figure 10 that the ganciclovir nanogel presents an interlaced network structure, and the presence of nanofibers in the skeleton can be observed, which is a gel network formed by the mutual crosslinking of nanofibers.

[0035] The ganciclovir nanofiber product of Example 4 was subjected to scanning electron microscope test, and the results are shown in Figure 11 , wherein, Figure 11 (a) is a scanning electron microscope image of the nanofiber, Figure 11 (b) is a particle size distribution graph of the nanofiber; it can be seen from Figure 11 that the prepared ganciclovir nanofiber is in the form of an elongated strip with a fiber width of 200-400 nm and an aspect ratio of 1.5-3.

[0036] II. Performance analysis of gelled and / or microparticulated nucleoside drugs The performances of the products obtained in Examples 1-4 are similar, and the product prepared in Example 1 is taken as an example to explain the performance of the product in detail.

[0037] The antioxidant performance of the lipoic acid grafted acyclovir prepared in Example 1 step (1) was investigated, and acyclovir was used as a comparison; the investigation items included DPPH clearance rate and ABTS clearance rate. Among them, the DPPH clearance rate test method is as follows: Mix 300 μL of 0.1 mM DPPH ethanol solution, 200 μL of 1 mg / mL drug solution and 2700 μL of absolute ethanol, test the absorbance (517 nm) at different times (every 5 min, a total of 30 min) by using a microplate reader, and calculate according to the following formula: Antioxidant capacity = [(Ab-As) / Ab] x 100%, wherein, Ab is the absorbance of DPPH ethanol solution, and As is the absorbance at each time point after drug treatment.

[0038] The ABTS clearance rate test method is as follows: Put 0.946 mg of potassium persulfate (K2S2O8) into 1 mL of 7 mM ABTS aqueous solution, mix thoroughly to obtain an ABTS ion solution, mix 100 μL of ABTS ion solution, 200 μL of 1 mg / mL drug solution and 2900 μL of deionized water, and then test the absorbance (714 nm) at different times (every 5 min, a total of 30 min) by using a microplate reader, and the calculation formula is the same as that of DPPH.

[0039] The 30-minute antioxidant performance test results of lipoic acid grafted acyclovir are shown in Figure 12 , wherein, Figure 12 (a) is the DPPH clearance rate, Figure 12 (b) is the ABTS clearance rate. It can be seen from Figure 12 that the DPPH radical clearance rate of lipoic acid grafted acyclovir reaches 60%, which is much higher than that of acyclovir (15%); the ABTS radical clearance rate of lipoic acid grafted acyclovir is 54%, which is much higher than that of acyclovir (13%), and the reason is that the disulfide bond in lipoic acid grafted acyclovir can react with active oxygen to achieve the purpose of clearing DPPH radicals and ABTS radicals.

[0040] The rheological test results of acyclovir nanogel of Example 1 are shown in Figure 13 , and it can be seen from Figure 13 that the storage modulus is higher than the loss modulus, indicating that acyclovir nanogel is successfully prepared.

[0041] The antioxidant performance test results of acyclovir nanogel of Example 1 are shown in Figure 14 , and the test method is the same as the antioxidant performance test method of lipoic acid grafted acyclovir above. It can be seen from Figure 14 that the DPPH radical clearance rate of acyclovir nanogel reaches 58%, and the ABTS radical clearance rate is 49%, which is basically consistent with the antioxidant performance of lipoic acid grafted drug, and both are much higher than that of the corresponding drug itself.

[0042] Although the present application has been described in detail with particular references to a presently preferred embodiment, it should be understood that the protection scope of the present patent is not limited to the specific embodiment described. Various modifications and changes can be made by those skilled in the art without departing from the protection scope of the present patent, as described in the claims.

Claims

1. A method for preparing a gelled and / or micronized nucleoside drug, characterized in that, The method comprises the following steps: S1: grafting lipoic acid to nucleoside drugs to obtain nucleoside drugs grafted with lipoic acid; S2: ring-opening polymerization of the nucleoside drugs grafted with lipoic acid to obtain gelled and / or micronized nucleoside drugs.

2. The production method according to claim 1, characterized by, The method for grafting lipoic acid to nucleoside drugs in S1 is as follows: lipoic acid, nucleoside drugs and esterification catalyst are dissolved in a reaction solvent, and stirred at room temperature for 12-24 h to obtain the product; the esterification catalyst is a mixture of 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4-dimethylaminopyridine.

3. The method of claim 2, wherein: The nucleoside drugs are acyclovir, the reaction solvent is dimethyl sulfoxide, the esterification catalyst is obtained by mixing 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4-dimethylaminopyridine at a molar ratio of 1:1; the molar ratio of lipoic acid, acyclovir and esterification catalyst is 1:1:3; the solid-liquid ratio of acyclovir to dimethyl sulfoxide is 1 mol:20 L.

4. The method of claim 3, wherein: The reaction time is 12 h.

5. The method of claim 2, wherein: The nucleoside drugs are ganciclovir, the reaction solvent is dimethyl sulfoxide, the esterification catalyst is obtained by mixing 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 4-dimethylaminopyridine at a molar ratio of 1:1; the molar ratio of lipoic acid, ganciclovir and esterification catalyst is 1:2:6; the solid-liquid ratio of ganciclovir to dimethyl sulfoxide is 1 mol:10 L.

6. The method of claim 5, wherein: The reaction time is 24 h.

7. The preparation method according to claim 1, characterized in that, The method for ring-opening polymerization in S2 is as follows: The nucleoside drugs grafted with lipoic acid are dispersed in an organic solvent to obtain a dispersion liquid with a concentration of 1-5 mg / mL; then the dispersion liquid is dropped into water at 80-100 ℃, and stirred and polymerized for 2-24 h to obtain the product.

8. The method of claim 7, wherein: The prepared drug is gelled nucleoside drugs; the concentration of the dispersion liquid is 5 mg / mL; and the stirring and polymerization time is 12 h.

9. The method of claim 7, wherein: The prepared drug is micronized nucleoside drugs; the concentration of the dispersion liquid is 1 mg / mL; and the stirring and polymerization time is 2 h.

10. The gelated and / or microparticulated nucleoside drug prepared according to the process of any one of claims 1 to 9, characterized in that: The micronized nucleoside drugs are nucleoside drug nanoparticles or nucleoside drug nanofibers; the particle size of the nucleoside drug nanoparticles is 200-350 nm; the width of the nucleoside drug nanofibers is 200-400 nm, and the length-width ratio is 1.5-3.

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