High-load isothiocyanate stable preparation and preparation method thereof

Abstract

The invention provides a high-load isothiocyanate stable preparation and a preparation method thereof, and belongs to the technical field of pharmaceutical preparations. According to the invention, the HPMC with a specific substitution degree is selected to prepare the isothiocyanate stabilizing preparation, the HPMC can stabilize the isothiocyanate through a hydrophobic methoxyl group, can also form hydrogen bonds with a trace amount of water in a system through hydrophilic hydroxyl and hydroxypropyl, and has a certain slow release effect. According to the invention, HPMC and isothiocyanate are uniformly dispersed in an aprotic solvent, stable combination of a hydrophobic isothiocyanate group in isothiocyanate and a methoxyl region of HPMC is realized, and an isothiocyanate stable preparation is obtained after drying. The isothiocyanate stable tablet with an ideal slow-release effect is prepared by adding auxiliary materials, namely microcrystalline cellulose, calcium carbonate and magnesium stearate.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical preparations, in particular to a high-load isothiocyanate stable preparation and a preparation method thereof. BACKGROUND

[0002] Isothiocyanate compounds are important natural active ingredients from Brassicaceae plants, mainly existing in mustard, horseradish, broccoli, cabbage and other plants, and are produced by glucosinolate enzymolysis. Isothiocyanate compounds have anti-inflammatory, antioxidant and anticancer properties and have attracted much attention in recent years. However, the stability of isothiocyanate is extremely poor, and the isothiocyanate group (-N=C=S) contained therein is easily inactivated due to hydrolysis, with a half-life of less than one day at 37℃. Therefore, there are few preparations with isothiocyanate as the single active ingredient on the domestic market at present, and the products containing some isothiocyanate compounds with plant powders such as broccoli and radish as the main components can be purchased, but the content of isothiocyanate is less than 1%.

[0003] In addition, isothiocyanate compounds have certain side effects, and direct intake may cause severe gastrointestinal irritation. CN113662935A discloses an oil agent for reducing the gastrointestinal reactions caused by sulforaphane and sulforaphen, which can improve the strong gastrointestinal irritation of isothiocyanate, but the drug loading of isothiocyanate is relatively low. Therefore, improving the stability and sustained-release property of isothiocyanate is crucial for promoting its clinical application. SUMMARY

[0004] Therefore, the purpose of the present application is to provide a high-load isothiocyanate stable preparation and a preparation method thereof. The isothiocyanate stable preparation provided by the present application has good stability, extremely high isothiocyanate loading rate, and takes into account the sustained-release effect.

[0005] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical solutions:

[0006] The present application provides an isothiocyanate stable preparation, comprising isothiocyanate and hydroxypropyl methyl cellulose loaded with the isothiocyanate.

[0007] In the hydroxypropyl methyl cellulose, the substitution degree of methoxy is 20-29%, and the substitution degree of hydroxypropyl is 7-12%.

[0008] Preferably, the isothiocyanate comprises one or more of sulforaphane, sulforaphen, allyl isothiocyanate, benzyl isothiocyanate and indole isothiocyanate.

[0009] Preferably, in the isothiocyanate stable preparation, the loading amount of isothiocyanate is 1-40%.

[0010] The application provides a preparation method of the isothiocyanate stable preparation, and comprises the following steps:

[0011] The isothiocyanate, the hydroxypropyl methyl cellulose and the aprotic solvent are mixed to obtain a mixed solution;

[0012] The aprotic solvent in the mixed solution is removed to obtain the isothiocyanate stable preparation.

[0013] Preferably, the mass ratio of the isothiocyanate to the hydroxypropyl methyl cellulose is 1:1.5-50.

[0014] The volume ratio of the aprotic solvent to the total mass of the isothiocyanate and the hydroxypropyl methyl cellulose is 5-50 mL:1 g.

[0015] Preferably, the mixing is stirring mixing, the stirring rate is 200-1000 rpm, and the stirring time is 10-240 min.

[0016] The application provides an application of the isothiocyanate stable preparation in preparation of an anti-inflammatory, anti-oxidation or anticancer drug.

[0017] The application provides an isothiocyanate stable tablet, which comprises the following components in mass fractions:

[0018]

[0019] The application provides a preparation method of the isothiocyanate stable tablet, and comprises the following steps:

[0020] The isothiocyanate stable preparation, the microcrystalline cellulose, the calcium carbonate and the magnesium stearate are stirred and mixed to perform tabletting to obtain the isothiocyanate stable tablet.

[0021] Preferably, the stirring rate is 3000-12000 rpm, and the stirring time is 1-60 min.

[0022] The tabletting pressure is 5-40 kN.

[0023] The present application provides a stable isothiocyanate preparation, comprising isothiocyanate and hydroxypropyl methylcellulose (HPMC) loaded with the isothiocyanate; in the hydroxypropyl methylcellulose, the substitution degree of methoxyl is 20-29%, and the substitution degree of hydroxypropyl is 7-12%. The hydroxypropyl methylcellulose has excellent biocompatibility and biodegradability. Due to the introduction of relatively hydrophobic methoxyl and hydrophilic hydroxypropyl on the glucose monomer, the HPMC not only has excellent water solubility, but also has good compatibility with hydrophobic compounds. The compatibility of HPMC with hydrophobic compounds means that the loading rate of isothiocyanate compounds is high, and the loading amount of isothiocyanate can be up to 40%. The present application uses HPMC as a dispersion carrier for isothiocyanate, and has the following advantages:

[0024] ①The water content is the main factor affecting the stability of isothiocyanate compounds. HPMC can not only stabilize isothiocyanate through hydrophobic methoxyl, but also form hydrogen bonds with trace water in the system through hydrophilic hydroxyl and hydroxypropyl, thereby reducing the content of free water in the preparation. These interactions spatially separate isothiocyanate and water, reducing the possibility of their reaction, thereby significantly improving the stability of isothiocyanate, and the t 0.9 (effective component remaining 90% time elapsed) is extended to 596.5 days.

[0025] ②HPMC is an excellent hydrophilic gel matrix material that can effectively control the release rate of the active ingredient. Isothiocyanate compounds have hydrophobic isothiocyanate (-N=C=S) groups and side chains, which can interact with the hydrophobic methoxyl region of HPMC with a specific substitution degree through hydrophobic interaction, thereby forming a stable high-loading solid dispersion with good sustained-release effect. When the loading amount of isothiocyanate is within 40%, the solid dispersion shows good sustained-release effect for isothiocyanate. The prerequisite for sustained release is the stable loading of HPMC for isothiocyanate, and then, due to the hydration of HPMC, a gel barrier is quickly formed on the surface, and isothiocyanate is slowly released by diffusing through the gradually swelling gel layer. According to the different substitution degrees and amounts, different release rates can be adjusted.

[0026] ③Compared with existing isothiocyanate capsule oil, the isothiocyanate stable preparation provided by the present application is a solid preparation, which has the advantages of easy storage and transportation.

[0027] ④The preparation method of the isothiocyanate stable preparation provided by the present application is simple in operation, low in cost, and easy to realize industrialized batch production. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1A photograph of the stable formulation of isothiocyanate obtained in Example 1;

[0029] Figure 2 An atomic force microscope image of the stable formulation of isothiocyanate obtained in Example 2;

[0030] Figure 3 A Raman mapping image of the stable formulation of isothiocyanate obtained in Example 2;

[0031] Figure 4 A graph of the accelerated stability test data of the stable formulation of isothiocyanate obtained in Example 2;

[0032] Figure 5 A graph of the in vitro release test data of the stable tablet of isothiocyanate obtained in Example 2;

[0033] Figure 6 A graph of the in vitro release test data of the stable tablet of isothiocyanate obtained in Comparative Example 1;

[0034] Figure 7 A photograph of the stable formulation of isothiocyanate obtained in Comparative Example 2;

[0035] Figure 8 A graph of the in vitro release test data of the stable tablet of isothiocyanate obtained in Comparative Example 3;

[0036] Figure 9 A photograph of the stable formulation of isothiocyanate obtained in Comparative Example 5. DETAILED DESCRIPTION

[0037] The present application provides a stable formulation of isothiocyanate, comprising isothiocyanate and hydroxypropyl methylcellulose on which the isothiocyanate is loaded.

[0038] In the hydroxypropyl methylcellulose, the degree of substitution of methoxyl group is 20 to 29%, and the degree of substitution of hydroxypropyl group is 7 to 12%.

[0039] In the present application, the isothiocyanate is an isothiocyanate compound present in a cruciferous plant, and preferably comprises one or more of sulforaphane, raphanusin, allyl isothiocyanate, benzyl isothiocyanate, and indole isothiocyanate.

[0040] In the present application, the degree of substitution of methoxyl in the hydroxypropyl methyl cellulose is 20-29%, specifically can be 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28% or 29%, preferably 28%; the degree of substitution of hydroxypropyl is 7-12%, specifically can be 7%, 8%, 9%, 10%, 11% or 12%, preferably 9%. By controlling the degree of substitution of methoxyl and hydroxypropyl in the hydroxypropyl methyl cellulose, the present application can ensure stable high loading of isothiocyanate compounds and appropriate release rate.

[0041] In the present application, the loading of isothiocyanate in the isothiocyanate stable preparation is 1-40%, specifically can be 1%, 5%, 10%, 15%, 20%, 30 or 40%. In the present application, the morphology of the isothiocyanate stable preparation is powder.

[0042] The present application provides a preparation method of the above-mentioned isothiocyanate stable preparation, comprising the following steps:

[0043] Mixing isothiocyanate, hydroxypropyl methyl cellulose and aprotic solvent to obtain a mixed solution;

[0044] Removing the aprotic solvent in the mixed solution to obtain the isothiocyanate stable preparation.

[0045] The present application mixes isothiocyanate, hydroxypropyl methyl cellulose and aprotic solvent to obtain a mixed solution. In the present application, the aprotic solvent is preferably a polar aprotic solvent, specifically can be dichloromethane and / or ethyl acetate. In the present application, the mass ratio of isothiocyanate to hydroxypropyl methyl cellulose is preferably 1:1.5-50, specifically can be 1:1.5, 1:4, 1:10, 1:20, 1:30, 1:40 or 1:50, more preferably 1:4; the volume ratio of the total mass of isothiocyanate and hydroxypropyl methyl cellulose to the aprotic solvent is preferably 1g:5-50mL, more preferably 1g:10mL.

[0046] In the present application, the mixing is preferably stirring mixing, the stirring rate is preferably 200-1000rpm, more preferably 400rpm; the stirring temperature is preferably 4-40℃, more preferably 25℃, and the time is preferably 10-240min, more preferably 30-60min. By the stirring mixing, the present application can make the hydroxypropyl methyl cellulose and isothiocyanate uniformly dispersed in the solvent.

[0047] After obtaining the mixed solution, the present application removes the aprotic solvent in the mixed solution to obtain the isothiocyanate stable preparation. In the present application, the removal of the aprotic solvent is preferably drying, which is preferably carried out in a vacuum drying oven, and the temperature of the drying is preferably 20-50℃, more preferably 25℃. The present application does not have special requirements for the time of the drying, and the solid is dried to a constant weight.

[0048] The present application provides the use of the above-mentioned isothiocyanate stable preparation in the preparation of anti-inflammatory, antioxidant or anticancer drugs.

[0049] The present application provides an isothiocyanate stable tablet, which comprises the following components in mass fraction:

[0050]

[0051]

[0052] In the present application, the microcrystalline cellulose acts as a filler for supporting the tablet structure.

[0053] In the present application, the calcium carbonate acts as a filler for improving the flowability.

[0054] In the present application, the magnesium stearate acts as a lubricant.

[0055] As a preferred scheme of the present application, the isothiocyanate stable tablet comprises the following components in mass fraction:

[0056]

[0057] In the present application, the thickness of the single isothiocyanate stable tablet is preferably 2-6mm, more preferably 3mm; and the mass is preferably 100-1000mg, more preferably 400mg.

[0058] The present application provides a preparation method of the above-mentioned isothiocyanate stable tablet, which comprises the following steps:

[0059] The isothiocyanate stable preparation, microcrystalline cellulose, calcium carbonate and magnesium stearate are stirred and mixed, and then tabletting is carried out to obtain the isothiocyanate stable tablet.

[0060] In this invention, the mixing is preferably dry mixing; the stirring and mixing rate is preferably 3000-12000 rpm, more preferably 8000 rpm; the time is preferably 1-60 min, more preferably 10 min. In this invention, the tableting pressure is preferably 5-40 kN, more preferably 8-10 kN.

[0061] In this invention, the isothiocyanate stabilized tablet is preferably spindle-shaped, with a length of 1-2 cm, more preferably 1.7 cm, and a width of 0.6-1.2 cm, more preferably 1 cm.

[0062] The following detailed description of the high-load isothiocyanate stabilizer and its preparation method provided by the present invention, with reference to the embodiments, should not be construed as limiting the scope of protection of the present invention.

[0063] Example 1

[0064] The degree of substitution of HPMC used in Example 1 was 28% methoxy and 9% hydroxypropyl, used to disperse sulforaphane, an isothiocyanate derived from broccoli.

[0065] The method for preparing isothiocyanate stabilizers comprises the following steps:

[0066] Weigh out sulforaphane and HPMC at a mass ratio of 1:4. Add dichloromethane at a total weight of 1 g to 10 mL solvent. Stir at 400 rpm for 30 minutes at 25 °C to ensure uniform dispersion of HPMC and sulforaphane in the solvent. Then, remove the solvent in a vacuum drying oven at 25 °C to obtain the isothiocyanate stabilized formulation with an isothiocyanate loading of 20%.

[0067] The method for preparing isothiocyanate stabilized tablets comprises the following steps:

[0068] (1) The prepared isothiocyanate stabilizer was mixed with excipients microcrystalline cellulose, calcium carbonate and magnesium stearate in a mass ratio of 1:2:0.25:0.03 and stirred at 8000 rpm for 10 minutes to obtain a uniform powder.

[0069] (2) The mixture powder was placed in a tablet press, each tablet containing 400 mg and a pressure of 8 kN, to obtain isothiocyanate stabilized tablets with a spindle shape, a length of 1.7 cm and a width of 1 cm.

[0070] A physical image of the isothiocyanate stabilizer obtained in Example 1 is shown below. Figure 1 As shown, by Figure 1 It can be seen that the obtained isothiocyanate stabilizer is a uniform white powder.

[0071] Example 2

[0072] The degree of substitution of HPMC used in Example 2 was 28% methoxy and 9% hydroxypropyl, and it was used to disperse radish-derived isothiocyanate raphanin.

[0073] The method for preparing isothiocyanate stabilizers comprises the following steps:

[0074] Weigh raphanin and HPMC separately according to a mass ratio of 1:4. Add dichloromethane according to a total weight of the mixture to solvent ratio of 1g:20mL. Stir at 400rpm for 20 minutes at 25℃ to disperse HPMC and raphanin evenly in the solvent. Then, remove the solvent in a vacuum drying oven at 25℃ to obtain the isothiocyanate stabilized formulation with an isothiocyanate loading of 20%.

[0075] The method for preparing isothiocyanate stabilized tablets comprises the following steps:

[0076] (1) The prepared isothiocyanate stabilizer was mixed with excipients microcrystalline cellulose, calcium carbonate and magnesium stearate in a mass ratio of 1:3:0.25:0.025 and stirred at 10000 rpm for 10 minutes to obtain a uniform powder.

[0077] (2) The mixture powder was placed in a tablet press, each tablet containing 200 mg and a pressure of 5 kN, to obtain isothiocyanate stabilized tablets with a spindle shape, a length of 1.7 cm and a width of 1 cm.

[0078] Atomic force microscopy image of the obtained isothiocyanate stabilized tablets is shown below. Figure 2 As shown, Figure 2 In the diagram, a represents HPMC, and b represents HPMC loaded with isothiocyanate. (From...) Figure 2 It can be seen that the surface roughness of HPMC increases after loading isothiocyanate.

[0079] The Raman profile of the obtained isothiocyanate stabilized tablets is shown in the figure. Figure 3 As shown, Figure 3 In the diagram, a represents the N=C bond distribution in the isothiocyanate group, b represents the C=S bond distribution in the isothiocyanate group, and c represents the CH bond distribution of the HPMC methyl group. Figure 3 It can be seen that the isothiocyanate group of isothiocyanate overlaps with the methoxy region of HPMC.

[0080] The accelerated stabilization experimental data of the obtained isothiocyanate stabilized tablets are shown in the figure below. Figure 4 As shown, accelerated stability testing was performed according to the Chinese Pharmacopoeia 2020 edition, 9001 Guidelines for Stability Testing of Drug Substances and Preparations. Figure 4 t can be calculated0.9 The time elapsed when 90% of the active ingredient remains is extended to 596.5 days.

[0081] The in vitro release experiment data graph of the obtained isothiocyanate stable tablet is shown in Figure 5 The in vitro release experiment was carried out according to the Chinese Pharmacopoeia 2020 version of simulated gastric juice and simulated intestinal juice. Figure 5 In the formula, a and b are the release times in the simulated gastric environment and intestinal environment, respectively. It can be seen from the formula that Figure 5 It can be seen that the release time of the isothiocyanate stable tablet in the simulated gastric environment and intestinal environment can be controlled at 2 hours and 6 hours, respectively.

[0082] Example 3

[0083] The degree of substitution of HPMC used in Example 3: methoxy is 25%, and hydroxypropyl is 8%, which is used to disperse isothiocyanate from broccoli source-sulforaphane.

[0084] The preparation method of the isothiocyanate stable preparation adopts the following steps:

[0085] According to the mass ratio of sulforaphane to HPMC 1:2, sulforaphane and HPMC are weighed, dichloromethane is added according to the ratio of the total weight of the mixture to the solvent 1g:10mL, and stirring is carried out at 500rpm for 30 minutes at 25°C to make HPMC and sulforaphane uniformly dispersed in the solvent. Then, the solvent is removed in a vacuum drying oven at 25°C to obtain an isothiocyanate stable preparation, and the loading capacity of isothiocyanate is 33.3%.

[0086] The preparation method of the isothiocyanate stable tablet adopts the following steps:

[0087] (1) The prepared isothiocyanate stable preparation is mixed with excipients microcrystalline cellulose, calcium carbonate, magnesium stearate and magnesium stearate according to the mass ratio 1:1.5:0.25:0.05, and a stirrer is used to stir at 8000rpm for 10 minutes to obtain a mixture powder;

[0088] (2) The mixture powder is placed in a tablet press, each tablet is 400 milligrams, and the pressure is 8kN to prepare an isothiocyanate stable tablet, which is spindle-shaped, 1.7cm in length and 1cm in width.

[0089] Comparative Example 1

[0090] Compared with Example 2, the difference is that the degree of substitution of HPMC used is: methoxy is 30%, and hydroxypropyl is 9%.

[0091] The isothiocyanate stable preparation and the isothiocyanate stable tablet are prepared in the manner of Example 2.

[0092] Figure 6In vitro release data graph for the isothiocyanate stable tablet obtained from Comparative Example 1. The data were obtained by Figure 6 It can be seen that the average release rate of Comparative Example 1 is 1 / 10 of that of Example 2, because the tablet of Comparative Example 1 forms a thick gel layer in the simulated gastrointestinal environment.

[0093] Comparative Example 2

[0094] The difference from Example 2 is the degree of substitution of the HPMC used: methoxyl 19%, hydroxypropyl 9%.

[0095] An isothiocyanate stable formulation was prepared in the manner of Example 2, and the physical appearance of the resulting formulation is shown in Figure 7 The results show that a uniform formulation could not be obtained, and the mixture exhibited an irregular block structure.

[0096] Comparative Example 3

[0097] The difference from Example 2 is the degree of substitution of the HPMC used: methoxyl 28%, hydroxypropyl 13%.

[0098] An isothiocyanate stable formulation and an isothiocyanate stable tablet were prepared in the manner of Example 2. The resulting tablet was loose in texture.

[0099] Figure 8 In vitro release data graph for the isothiocyanate stable tablet obtained from Comparative Example 3. The data were obtained by Figure 8 It can be seen that the tablet has little sustained release effect in the simulated gastrointestinal environment, and the loaded sulforaphane is completely released within 3 minutes.

[0100] Comparative Example 4

[0101] The difference from Example 2 is the degree of substitution of the HPMC used: methoxyl 28%, hydroxypropyl 6%.

[0102] An isothiocyanate stable formulation and an isothiocyanate stable tablet were prepared in the manner of Example 2. The resulting tablet completely releases sulforaphane in the simulated gastrointestinal environment in 18 hours and 24 hours, respectively, and the release rate is slow, and the effective ingredient utilization rate is low.

[0103] Comparative Example 5

[0104] The difference from Example 1 is that the mass ratio of sulforaphane to HPMC is 1:1.4.

[0105] An isothiocyanate stable formulation was prepared in the manner of Example 1, and the results were a mixture of irregular sulforaphane and HPMC that was bonded into a block, and the physical appearance of the resulting formulation is shown in Figure 9

[0106] ​The above merely describes the preferred embodiments of the present application, and it should be pointed out that, for those skilled in the art, several improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as falling within the protection scope of the present application.

Claims

1. A stable formulation of isothiocyanate, characterized in that, The isothiocyanate and hydroxypropyl methyl cellulose loaded with the isothiocyanate; The degree of substitution of the methoxyl group in the hydroxypropyl methyl cellulose is 20-29%, and the degree of substitution of the hydroxypropyl group is 7-12%.

2. The isothiocyanate stable formulation of claim 1, wherein, The isothiocyanate includes one or more of sulforaphane, raphanusulfane, allyl isothiocyanate, benzyl isothiocyanate and indole isothiocyanate.

3. The isothiocyanate stable formulation according to claim 1 or 2, characterized in that, The loading amount of the isothiocyanate in the isothiocyanate stable preparation is 1-40%.

4. A process for the preparation of the stable preparation of isothiocyanate according to any one of claims 1 to 3, characterized in that, The method comprises the following steps: The isothiocyanate, the hydroxypropyl methyl cellulose and the aprotic solvent are mixed to obtain a mixed solution; The aprotic solvent in the mixed solution is removed to obtain the isothiocyanate stable preparation.

5. The preparation method according to claim 4, characterized in that, The mass ratio of the isothiocyanate to the hydroxypropyl methyl cellulose is 1:1.5-50; The volume ratio of the aprotic solvent to the total mass of the isothiocyanate and the hydroxypropyl methyl cellulose is 1g:5-50mL.

6. The preparation method according to claim 4, characterized in that, The mixing is stirring mixing, the stirring rate is 200-1000rpm, and the stirring time is 10-240min.

7. Use of the isothiocyanate stable preparation of any one of claims 1-3 or the isothiocyanate stable preparation prepared by the preparation method of any one of claims 4-6 in preparation of an anti-inflammatory, antioxidant or anticancer drug.

8. An isothiocyanate stable tablet characterized in that, The components include the following mass fractions: The isothiocyanate stable preparation is the isothiocyanate stable preparation of any one of claims 1-3 or the isothiocyanate stable preparation prepared by the preparation method of any one of claims 4-6.

9. The process for the preparation of isothiocyanate stable tablets according to claim 8, characterized in that, The method comprises the following steps: The isothiocyanate stable preparation, microcrystalline cellulose, calcium carbonate and magnesium stearate are stirred and mixed to obtain isothiocyanate stable tablets.

10. The method of claim 9, wherein, The stirring rate is 3000-12000rpm, and the time is 1-60min. The pressure for tabletting is 5-40kN.

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