Composition for increasing solubility of quinone compound and application thereof

Abstract

The invention discloses a composition for increasing solubility of quinone compounds and application thereof, and belongs to the technical field of pharmaceutical preparations, the composition comprises insoluble quinone compounds, amphiphilic phospholipids or derivatives thereof and / or amphiphilic stabilizers, and the insoluble quinone compounds are benzoquinone and naphthoquinone compounds; wherein the benzoquinone compound is selected from idebenone, coenzyme Q10, 2, 6-dimethoxy-1, 4-benzoquinone, 2-methoxy-6-pentyl-1, 4-benzoquinone, 2, 3-dimethoxy-5-methyl-6-decaisopentenyl-1, 4-benzoquinone and creptyquinone, and the benzoquinone compound is selected from idebenone, coenzyme Q10, 2, 6-dimethoxy-1, 4-benzoquinone, 2, 3-dimethoxy-5-methyl-6-decaisopentenyl-1, 4-benzoquinone, 2, 3-dimethoxy-5-methyl-6-decaisopentenyl-1, 4-benzoquinone and creptyquinone. The naphthoquinone compound is selected from alkannin and derivatives thereof, juglone, blue snow quinone and isoalkannin. The composition has the advantages of strong solubilizing ability and good stability, and when the drug concentration is greater than 0.5%, the phenomenon of drug crystallization and precipitation does not occur within 6 months in an acceleration test at 40 DEG C and under the condition that the composition is diluted by 50 times. The composition can be used for preparing a preparation for topical administration through skin or mucous membranes or systemic administration through injection and gastrointestinal tracts.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a composition for increasing the solubility of quinone compounds, a preparation method and application thereof. Background Art

[0002] Quinone compounds are a class of small molecule compounds existing in nature. Most of them contain unsaturated cyclic diketone structures and have a wide range of pharmacological activities. Many quinone compounds have low solubility. Representative hydrophobic para-benzoquinone compounds include idebenone, coenzyme Q10, and oxadiazine quinone, and representative hydrophobic naphthoquinone compounds include shikonin and isoshikonin. Low solubility seriously limits the clinical application of such active compounds. Improving the local or systemic absorption of poorly soluble compounds has always been a major challenge in the field of formulation technology. Due to their poor solubility, hydrophobic quinone compounds have problems of poor absorption and low bioavailability when administered via different routes. Therefore, it is necessary to use appropriate formulation technology to increase solubility, so as to avoid limiting their absorption and clinical application due to factors such as insufficient dissolution of the compound.

[0003] With the development of formulation technology, new technologies represented by nanoemulsions, inclusion compounds, micelles, etc. are used to improve the solubility of poorly soluble compounds. However, the above technologies still face some difficult-to-solve obstacles, such as: 1. Nanoemulsions use oil phases to increase the solubility of poorly soluble drugs. There is an oil-water two-phase interface, and a large amount of surfactants and auxiliary surfactants are required to maintain its thermodynamic stability, which is likely to cause irritation; 2. Nanoemulsions have poor physical stability, and structural damage will occur during storage or after administration, resulting in drug molecule leakage and aggregation, resulting in decreased solubility and permeability; 3. Inclusion compounds represented by β-cyclodextrin have structural selectivity for solubilization of poorly soluble drug molecules, and the solubilization effect on poorly soluble compounds with quinone structures is very limited.

[0004] Micelles are a very promising solubilization technology, but one of the main technical obstacles faced by micellar solubilization technology is the poor stability after loading drugs, which is manifested as the rapid crystallization of drug molecules in the drug-loaded micelle solution during dilution or placement, resulting in a decrease in solubility and failure to meet the requirements of medication. The stability of drug-loaded micelles is closely related to the drug loading amount. As the drug loading amount increases, the stability of micelles decreases significantly. In order to solve the stability problem, the micelle solution needs to be freeze-dried and stored, and needs to be re-dissolved and prepared before use, such as the paclitaxel micelles currently used in clinical practice. When the drug concentration in the micelle is low, the stability of the micelle solution may be enhanced. For example, the cyclosporine A micelle eye drops (Cequa) used clinically exist in liquid form, and its drug concentration is only 0.09%. At present, there is a lack of general technical means for increasing the stability of high-loaded drug micelle solutions in this technical field. Summary of the invention

[0005] In order to overcome the defects in the prior art, the inventors unexpectedly discovered that bile salt amphiphilic stabilizers and compositions thereof with amphiphilic phospholipids or their derivatives can significantly improve the solubility of poorly soluble p-benzoquinone and naphthoquinone compounds, and that the stability of the solubilized solution is good when the drug concentration is greater than 0.5% (50 times higher than the drug concentration of commercially available cyclosporine A micelles), and that the micelle solution does not precipitate drug molecules when placed at high temperature (40° C.) for 6 months.

[0006] On this basis, the purpose of the present invention is to provide a poorly soluble quinone compound solubilization composition and its application. The quinone compound solubilization composition prepared by the present invention has a particle size of less than 100nm, good stability, and high drug concentration, which provides a solution for the preparation of such drugs into dosage forms suitable for oral, injection and external use for clinical application.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a solubilizing composition for increasing the solubility of a poorly soluble quinone compound, wherein the solubilizing composition comprises a poorly soluble quinone compound and an amphiphilic stabilizer, and may further contain an amphiphilic phospholipid or a derivative thereof.

[0009] As a preferred embodiment, in the solution of the solubilizing composition, the poorly soluble quinone compounds are preferably benzoquinone and naphthoquinone compounds; wherein the benzoquinone compounds are selected from: idebenone, coenzyme Q10, 2,6-dimethoxy-1,4-benzoquinone, 2-methoxy-6-pentyl-1,4-benzoquinone, 2,3-dimethoxy-5-methyl-6-decisopentenyl-1,4-benzoquinone and shikonin; the naphthoquinone compounds are selected from: shikonin and its derivatives, juglone, blue snow quinone and isoshikonin.

[0010] As a preferred mode, when the solution of the solubilizing composition contains amphiphilic phospholipids or their derivatives and amphiphilic stabilizers, the amphiphilic phospholipids or their derivatives and the amphiphilic stabilizers self-assemble to form stable nano-micelle particles, and when only the amphiphilic stabilizers are contained, the amphiphilic stabilizers self-assemble to form stable nano-micelle particles. Preferably, the poorly soluble quinone compounds are stably solubilized in the solution of the solubilizing composition, and the drug molecules do not precipitate during the storage process.

[0011] As a preferred mode, in the solution of the solubilizing composition, the amphiphilic phospholipid or its derivative is preferably a glycerophospholipid or its derivative, including a phospholipid molecule having the following structural characteristics: two hydroxyl groups of glycerol form esters with fatty acids, and the third hydroxyl group is phosphated. Further preferably, the glycerophospholipid or its derivative is selected from one or more of natural phospholipids, semi-synthetic phospholipids, and synthetic phospholipids; the natural phospholipid is selected from one or more of soybean lecithin, egg yolk lecithin, cardiolipin, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylinositol; the semi-synthetic phospholipid is selected from one or more of hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, and hydrogenated sunflower lecithin; the synthetic phospholipid is selected from one or more of synthetic glycerophospholipids and polyethylene glycol phospholipids. Further preferably, the amphiphilic phospholipid or its derivative is selected from one or more of soybean lecithin, egg yolk lecithin, hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, sn-1 and sn-2 stearoylation, palmitoylation, oleoylation, myristoylated choline, sn-1 and sn-2 stearoylation, palmitoylation, oleoylation, myristoylated glycerol, sn-1 and sn-2 stearoylation, palmitoylation, oleoylation, myristoylated ethanolamine.

[0012] As a preferred embodiment, in the solution of the solubilizing composition, the amphiphilic stabilizer is preferably bile salt or its derivatives, further preferably bile acid and its salts, bile acid-amino acid conjugates and their salts, more preferably glycocholic acid and its salts, deoxycholic acid and its salts, glycodeoxycholic acid and its salts, taurocholic acid and its salts, taurodeoxycholic acid and its salts.

[0013] As a preferred embodiment, the solution of the solubilization composition can be mixed with other pharmaceutical excipients or other substances before use.

[0014] As a preferred embodiment, the solution of the solubilizing composition can be stored after removing the solvent through a suitable drying process, and then used after being re-dissolved.

[0015] As a preferred mode, in the solution of the solubilizing composition, the particle size range of the nanoparticles formed by the amphiphilic stabilizer or the self-assembly of the nanoparticles with the amphiphilic phospholipid or its derivative is less than 100 nm. Preferably, the particle size range of the nanoparticles is less than 40 nm, and more preferably, the particle size range of the nanoparticles is less than 20 nm.

[0016] As a preferred mode, in the solution of the solubilizing solution composition, the concentration of the amphiphilic stabilizer is 0.5-80%, preferably 1%-60%, by weight percentage; the mass ratio of the amphiphilic phospholipid or its derivative to the amphiphilic stabilizer is between 0:1-4:1, preferably 0:1-3:1. The concentration of the poorly soluble quinone compound is 0.01%-25%, preferably 0.5-20%.

[0017] In the second aspect, the present invention provides that the solubilizing composition described in the first aspect can be used to prepare various preparation forms for gastrointestinal and parenteral administration, and the parenteral administration route includes local administration through the skin or mucosa, and systemic administration via injection.

[0018] In a third aspect, the present invention provides the solubilization composition described in the first aspect for use in local skin beauty, disease prevention, treatment or auxiliary treatment.

[0019] In a fourth aspect, the present invention provides use of the solubilizing composition described in the first aspect in preparing a drug delivery system.

[0020] The solubilized composition of the present invention is a colloidal delivery system, and the quinone compounds are solubilized in the hydrophobic core of the composition delivery system.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The composition for solubilizing water-insoluble quinone compounds of the present invention has the advantages of strong solubilizing ability and good stability. When the drug concentration is greater than 0.5%, no drug crystal precipitation occurs under high temperature (40°C) accelerated test for 6 months and under the condition of 50 times dilution. The most significant advantage of the present invention is that the water-insoluble quinone compound solubilizing solution composition can be used to prepare suitable preparations, which can be administered topically through the skin or mucous membrane or systemically through injection, gastrointestinal tract, and inhalation. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the representative particle size measurement result of Sample 2 in Example 1 of the present invention.

[0024] Figure 2 This is a transmission electron microscope photograph of sample 2 in Example 1 of the present invention.

[0025] Figure 3 This is the cumulative transdermal permeation curve of Sample 2 and Idebenone solution in Example 2 of the present invention within 24 hours. DETAILED DESCRIPTION

[0026] The present invention will be further described with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0027] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0028] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0029] Example 1: Preparation of composition solution and determination of pharmaceutical properties

[0030] Weigh the required glycerophospholipids or their derivatives and poorly soluble quinone compounds according to the weight percentages in Table 1-2, add anhydrous ethanol solution or other organic solvent, stir until a clear solution is obtained, heat and evaporate the solvent under vacuum conditions, add water or phosphate buffered saline solution of a stabilizer, mix and stir evenly to obtain a solution of the drug-loaded micelle composition.

[0031] Take the solutions of the composition of sample 2, sample 6 and sample 8-10, dilute them 50 times with water, observe whether there is precipitation, and determine the concentration and encapsulation efficiency of the drug after filtering the diluted solution through a 0.1 μm microporous filter membrane. The results are shown in Table 3. Take appropriate amounts of the solutions of the composition of sample 2, sample 6 and sample 8-10, fill them into vials, cap them and place them at 40°C ± 2°C and RH75% ± 5% for 6 months, take samples at 0, 3, and 6 months, place them at 30°C ± 2°C and RH65% ± 5% for 24 months, take samples at 12 and 24 months, observe the appearance and determine the concentration and encapsulation efficiency of the drug. The results are shown in Table 4.

[0032] Table 1 Solution formulation composition of phospholipid / cholate derivative composition

[0033]

[0034] Table 2 Solution formulation composition of the phospholipid / cholate derivative composition of the comparative example

[0035]

[0036] Table 3 Main pharmaceutical properties of samples 1-10

[0037]

[0038]

[0039] a: drug concentration in micellar solution; b: drug solubility in water.

[0040] The representative particle size measurement results of sample 2 are as follows: Figure 1 shown.

[0041] The transmission electron microscope photo of sample 2 is shown in Figure 2 shown.

[0042] The results in Table 3 show that the average light intensity particle size of the micellar nanoparticles prepared by the present invention is less than 100 nm. Compared with the solubility in water, the micelles of the composition of bile salt and phospholipids or their derivatives can significantly increase the solubility of quinone compounds. Compared with the comparative example, the mass ratio of bile salt to phospholipid in the composition is greater than or equal to 1:4, and the drug loading and drug concentration of the micellar solution are significantly increased.

[0043] Table 4 Stability results of sample 2, sample 6 and samples 8-10 solutions

[0044]

[0045]

[0046] As shown in Table 4, when the mass ratio of sodium glycocholate to phospholipid in the composition is less than or equal to 1:5 (samples 9 and 10), the solubilized micelles show drug crystallization and decreased encapsulation efficiency under dilution conditions and stability test conditions, indicating that the stability of the micelle composition is poor. Under the same conditions, when the mass ratio of sodium glycocholate to phospholipid is greater than or equal to 1:4, the appearance and encapsulation efficiency of the solubilized micelle solution do not change significantly, showing better stability.

[0047] Example 2: Study on topical skin administration of the composition solution

[0048] Preparation of ex vivo mouse skin

[0049] The abdominal skin of healthy male rats was used as the model skin. The preparation process was as follows: anesthetize with intraperitoneal injection of 20% urethane aqueous solution, remove the long hair on the abdomen with a pet electric razor, and then remove the remaining short hair with a razor. Kill by dislocating the neck, peel off the abdominal skin, and carefully remove the subcutaneous fat and connective tissue. Rinse with saline and set aside.

[0050] In vitro permeation test

[0051] The transdermal test was conducted using a horizontal transdermal diffusion cell at 32°C ± 0.5°C. The skin was fixed between the two halves of the horizontal diffusion cell, with the stratum corneum facing the supply cell and the dermis facing the receiving cell. The effective permeation area (S) of the diffusion cell was 2.0 cm 2, the volume of the receiving pool is 20 ml. The same volume of sample 2 and idebenone solution (ethanol / water = 4 / 3, v / v) were added to the supply chamber, and the electromagnetic stirrer was turned on to stir at a speed of 100 r / min. 1 ml of the receiving solution was taken out at 1, 2, 3, 4, 5, 6, 7, 8, 10, and 12 hours respectively (an equal amount of blank receiving solution was added after each sampling). After the obtained sample was centrifuged, 20 μl of the supernatant was injected and the drug content was determined by HPLC. The chromatographic conditions were: kromasil c18 (200×4.6 mm), mobile phase: methanol: water (78:22, v / v), detection wavelength: 278 nm, flow rate: 1.0 ml / min, injection volume: 20 μl, column temperature: 30°C. The cumulative permeation amount was plotted against time to draw the transdermal permeation curve.

[0052] The experimental results are as follows Figure 3 As shown in Figure 2, the cumulative percutaneous permeation of idebenone solution and sample 2 in 24 hours was 203.90±12.66μg / cm 2 , 195.94±15.20μg / cm 2 The results showed that the percutaneous penetration effect of the prepared high-concentration idebenone composition was similar to that of the idebenone solution. Since the solution contained a higher concentration of ethanol, it was more irritating to the skin. Compared with the solution, the idebenone composition micelle was more suitable for use as an external preparation.

[0053] Example 3: Oral and intravenous administration study of the composition solution

[0054] Male Wistar rats (190-210 g) were used as model animals. Idebenone solution and sample 6 (equivalent to 75 mg / kg of idebenone) were administered orally or intravenously. About 0.5 mL of blood was collected from the eye sockets at 0.5, 1, 2, 3, 4, 6, 8, 12, and 24 h. Centrifugation was performed at 16,000 rpm and 4°C for 5 min, and the upper plasma sample was stored in a -80°C refrigerator. After all samples were collected, the idebenone concentration in plasma was determined by liquid chromatography-mass spectrometry. Chromatographic conditions: Phenomenex Luna C18 (2.0×50 mm, particle size 3 μm) was used, the mobile phase was 5 mM ammonium acetate-methanol (25:75, v / v), and the flow rate was 0.3 mL min -1, injection volume was 5 μl, column temperature was 35°C. Mass spectrometry conditions: APCI source was used as ion source, positive ion monitoring, scanning mode was multiple reaction monitoring (MRM), Gas1 (N2): 50 psi, Curtain Gas (N2): 40 psi, CAD Gas (N2): 7 psi, TEM: 500°C, Nebulizer Current: 3 μA. Ion detection channel: Idebenone (analyte): m / z 339→m / z 339, Duloxetine (internal standard): m / z 298.2→m / z 154.1.

[0055] The experimental results are shown in Table 5 and Table 6. The absolute bioavailability of the idebenone solution and sample 6 after oral administration is 43.39% and 45.98%, respectively. The absorption degree of the prepared composition solution is similar to that of the solution, indicating that the idebenone composition solution has good stability in vivo, and no drug crystals are precipitated after dilution with digestive fluid or blood. There is no significant difference in the pharmacokinetic curve and main kinetic parameters of the idebenone solution and sample 6 after intravenous administration. Compared with the idebenone solution containing ethanol, sample 6 (the micellar solution of the idebenone composition) does not contain an organic solvent and is less irritating to oral and injection administration.

[0056] Table 5 Pharmacokinetic parameters of sample 6 and idebenone solution for oral administration

[0057]

[0058] Table 6 Pharmacokinetic parameters of sample 6 and idebenone solution administered intravenously

[0059]

Claims

1. A composition for increasing the solubility of quinone compounds, It is characterized in that The composition comprises a poorly soluble quinone compound and an amphiphilic stabilizer.

2. A composition for increasing the solubility of quinone compounds according to claim 1, It is characterized in that It also contains amphiphilic phospholipids or their derivatives.

3. A composition for increasing the solubility of quinone compounds according to claim 1, It is characterized in that The poorly soluble quinone compounds are benzoquinone and naphthoquinone compounds; wherein the benzoquinone compounds are selected from: idebenone, coenzyme Q10, 2,6-dimethoxy-1,4-benzoquinone, 2-methoxy-6-pentyl-1,4-benzoquinone, 2,3-dimethoxy-5-methyl-6-decisopentenyl-1,4-benzoquinone and shikonin; the naphthoquinone compounds are selected from: shikonin and its derivatives, juglone, blue snow quinone and isoshikonin.

4. A composition for increasing the solubility of quinone compounds according to claim 1, It is characterized in that The amphiphilic stabilizer is selected from cholic acid and its salts, cholic acid-amino acid conjugates and their salts.

5. A composition for increasing the solubility of quinone compounds according to claim 4, It is characterized in that The amphiphilic stabilizer is selected from glycocholic acid and its salts, deoxycholic acid and its salts, glycodeoxycholic acid and its salts, taurocholic acid and its salts, and taurodeoxycholic acid and its salts.

6. A composition for increasing the solubility of quinone compounds according to claim 1 or 2, It is characterized in that When the solution of the composition contains amphiphilic phospholipids or their derivatives and amphiphilic stabilizers, the amphiphilic phospholipids or their derivatives and the amphiphilic stabilizers self-assemble to form stable nano-micelle particles; when only the amphiphilic stabilizer is contained, the amphiphilic stabilizer self-assembles to form stable nano-micelle particles, and the particle size of the nanoparticles is less than 100 nm; the poorly soluble quinone compounds are stably solubilized in the solution of the composition, and the drug molecules do not precipitate during being placed under the conditions of 40°C±2°C and RH75%±5% for 6 months.

7. A composition for increasing the solubility of quinone compounds according to claim 2, It is characterized in that The amphiphilic phospholipids or their derivatives are preferably selected from one or more of natural phospholipids, semi-synthetic phospholipids, and synthetic phospholipids; the natural phospholipids are selected from one or more of soybean lecithin, egg yolk lecithin, cardiolipin, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylinositol; the semi-synthetic phospholipids are selected from one or more of hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, and hydrogenated sunflower phospholipids; the synthetic phospholipids are selected from one or more of synthetic glycerophospholipids and polyethylene glycol phospholipids.

8. A composition for increasing the solubility of quinone compounds according to claim 2, It is characterized in that In the solution of the composition, the concentration of the amphiphilic stabilizer is 0.5-80% by weight, the mass ratio of the amphiphilic phospholipid or its derivative to the amphiphilic stabilizer is between 0:1-4:1, and the concentration of the poorly soluble quinone compound is 0.01%-25%.

9. Use of the composition for increasing the solubility of quinone compounds according to any one of claims 1 to 5 in preparing a drug delivery system.

10. Use of the composition for increasing the solubility of quinone compounds according to any one of claims 1 to 5 in preparing various preparations for gastrointestinal and parenteral administration.

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