Patent Application
20190336479
The present invention belongs to the technical field of medicine, particularly, relates to a Tecovirimat-containing pharmaceutical composition for injection and a preparation method thereof.
Smallpox is a highly lethal and infectious disease, which is mainly spread by droplet or clothing, and the clinical manifestations of which are facial and systemic rashes until death. Although WHO declared in 1980 that smallpox had been eliminated in the nature, smallpox will still have disastrous consequences for humans or even cause wars around the world once it is unintentionally or deliberately released. Since smallpox vaccines have serious adverse reactions, medical therapy is still necessary. However, up to now, no therapeutic regimen against smallpox has been approved worldwide.
Tecovirimat (designated as ST-246, with a chemical name of 4-trifluoromethyl-N-(3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl)-benzamide, Formula 1) is a highly active small-molecule virus inhibitor, which works by binding to viral genes so as to prevent viral release in a cell. In addition, the use of Tecovirimat and a smallpox vaccine in combination can also prevent and treat the adverse reactions caused by the smallpox vaccine, reduce the damage level and promote wound healing. However, Tecovirimat has a very poor water solubility, which is less than 3 μg/ml, and its low solubility restricts its clinical application.
CN201180048043.1 discloses a novel liquid preparation in which Tecovirimat is solubilized in cyclodextrin and a novel method for preparing the preparation. In the method, the solubility of Tecovirimat (ST-246) is increased to 1.5-11 mg/ml by using 20%-40% (w/v) hydroxypropyl-cyclodextrin (HP-3-CD) at 37° C. In particular, Tecovirimat (ST-246) can have a maximal solubility of 21.23 mg/ml by using 40% (w/v) HP-β-CD at 70° C. The presence of a cosolvent (PEG400) and a nonionic surfactant (Tween 80) cannot further improve the solubility of ST-246 in HP-β-CD.
Contents of Invention
After research, the inventors have creatively invented a novel ternary solubilizing composition comprising Tecovirimat, cyclodextrin, and an additive Meglumine (MEG), which greatly improves the solubility (with a maximal solubility of up to 152 mg/ml) of the Tecovirimat with poor solubility as compared with the prior art, and has the advantages such as simple formulation process, short preparation time, stable quality, strong controllability, good reproducibility and low cost.
The present invention includes the following items:
1. A pharmaceutical composition of Tecovirimat for injection, comprising Tecovirimat as an active ingredient, cyclodextrin and an additive,
wherein said additive is selected from the group consisting of meglumine, glycine, arginine, hydroxypropyl methyl cellulose, polyethylene glycol, chitosan, and polyvinyl pyrrolidone, and is preferably meglumine.
2. The pharmaceutical composition according to Item 1 of the present invention, wherein said cyclodextrin is selected from the group consisting of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin and a pharmaceutically acceptable cyclodextrin derivative (e.g. dimethyl-β-cyclodextrin, 2-hydroxyethyl-β-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, 3-hydroxypropyl-β-cyclodextrin, trimethyl-β-cyclodextrin);
preferably, said cyclodextrin is selected from the group consisting of β-cyclodextrin, dimethyl-β-cyclodextrin, 2-hydroxyethyl-β-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, 3-hydroxypropyl-β-cyclodextrin, and trimethyl-β-cyclodextrin;
further preferably, said cyclodextrin is selected from the group consisting of 2-hydroxypropyl-β-cyclodextrin, and 3-hydroxypropyl-β-cyclodextrin;
further more preferably, said cyclodextrin is 2-hydroxypropyl-β-cyclodextrin.
3. The pharmaceutical composition according to Item 1 or Item 2 of the present invention, wherein said cyclodextrin and Tecovirimat have a weight ratio of 4˜10:1, preferably 5˜8:1, further preferably 5:1, 6:1, 7:1, or 8:1, more preferably 6:1.
4. The pharmaceutical composition according to any one of Items 1 to 3 of the present invention, wherein said additive and Tecovirimat have a weight ratio of 0.5˜5:1, preferably 1˜4:1, further preferably 1:1, 1.6:1, 2:1, 3:1, or 4:1, more preferably 2:1 or 1.6:1.
5. The pharmaceutical composition according to any one of Items 1 to 4 of the present invention, wherein said additive and cyclodextrin have a weight ratio of 1:1˜5, preferably 1: 2˜4, further preferably 1:2.5, 1:3, 1:3.5, or 1:3.75.
6. The pharmaceutical composition according to any one of Items 1 to 5 of the present invention, wherein said pharmaceutical composition further comprises one or more components selected from the group consisting of water, glucose, and physiological saline.
7. The pharmaceutical composition according to any one of Items 1 to 6 of the present invention, wherein said pharmaceutical composition is a liquid, wherein Tecovirimat preferably has a concentration of 15˜80 mg/ml, more preferably has a concentration of 50 mg/ml.
8. The pharmaceutical composition according to any one of Items 1 to 7 of the present invention, wherein an amount of said hydroxypropyl-J3-cyclodextrin is 10% (w/v)-40% (w/v), preferably 30% (w/v).
9. A method for preparing the pharmaceutical composition according to any one of Items 1 to 8 of the present invention, comprising the following steps:
a) dissolving an additive and cyclodextrin in a desired volume of water, and mixingwell;
b) adding Tecovirimat, and stirring well;
c) carrying out sterilization,
optionally, sterilization is followed by drying, to obtain the desired product.
10. The method for preparing the pharmaceutical composition according to Item 9 of the present invention, wherein said drying is freeze-drying.
11. A method for treating smallpox, comprising administering to a subject in need thereof a therapeutically and/or prophylactically effective amount of the pharmaceutical composition according to any one of Items 1 to 8 of the present invention.
12. The pharmaceutical composition according to any one of Items 1 to 8 of the present invention, for use in the treatment of smallpox.
13. Use of the pharmaceutical composition according to any one of Items 1 to 8 of the present invention for the manufacture of a medicament for treating smallpox.
In a preferred embodiment, the pharmaceutical composition according to the present invention can be prepared by the following method comprising:
a. dissolving an additive and cyclodextrin in a desired volume of water, and mixing well;
b. adding the active ingredient Tecovirimat, and stirring in a water bath;
c. subjecting the prepared solution to aseptic filtration or autoclaving;
d. subpackaging the resultant solution into vials or ampoules, which are then sealed/capped.
In another preferred embodiment, the pharmaceutical composition according to the present invention can also be prepared by another method comprising:
a. dissolving an additive and cyclodextrin in a desired volume of water, and mixing well;
b. adding the active ingredient Tecovirimat, and stirring in a water bath;
c. subjecting the prepared solution to freeze-drying to remove water,
d. subpackaging the resultant solution into vials or ampoules, which are then sealed/capped.
In the present invention, the term “an additive” refers to a substance that can interact with an active ingredient, so as to enhance the inclusion efficiency of cyclodextrin, and further to improve drug solubility.
In the present invention, “% (w/v)” refers to a mass/volume concentration, which represents the mass (expressed as gram) of a solute contained in per 100 ml of a solution. For example, 20% (w/v) represents 20 g of a solute contained in per 100 ml of a solution.
The embodiments of the present invention are described in detail by combining the following examples. However, a person skilled in the art will understand that the following examples and experimental examples are only used to describe the present invention, and should not be regarded as defining the scope of the present invention. In the case where the concrete conditions are not indicated in the examples and experimental examples, the examples are carried out according to conventional conditions or the conditions recommended by manufacturers. The reagents or apparatuses, the manufacturers of which are not indicated, are the conventional products that are commercially available.
Preparation method: a prescribed amount of meglumine and hydroxypropyl-β-cyclodextrin were dissolved in a certain volume of water, and then a prescribed amount of Tecovirimat was added; water was added to a final volume of 100 ml, and the resultant mixture was stirred at a medium speed at 60° C. in a water bath, thereby obtaining the desired composition.
Preparation method: a prescribed amount of meglumine and hydroxypropyl-3-cyclodextrin were dissolved in a certain volume of water, and then a prescribed amount of Tecovirimat was added; water was added to a final volume of 100 ml, and the resultant mixture was stirred at a medium speed at 60° C. in a water bath, thereby obtaining the desired composition.
Experimental method: the solubilizing composition for injection (4 ml) as prepared in Example 1 was added to 100 ml glucose injection or physiological saline, and changes of the appearance, pH value, and the related substance were observed at 0 h, 2 h, 4 h, 6 h, and 8 h, respectively.
The experimental results were shown in Table 3 and Table 4.
The experimental results show that the solubilizing composition had good compatibility with glucose injection or physiological saline.
Experimental method: to 10 test tubes, designated as 1˜10, 1 ml phosphate buffer (pH 7.4) was added, and to Tube 1, the solubilizing composition (1 ml) as prepared in Example 1 or Example 2 was then added; the resultant solution was mixed well under shaking, the resultant solution (1 ml) was taken from Tube 1, and added to Tube 2; the operation was repeated until Tube 10, and the tubes were on standing and were observed for clarity.
Experimental result: no drug was precipitated in 10 tubes within 5 min, indicating that the formulation had good compatibility with the phosphate buffer (pH=7.4).
An excessive amount of Tecovirimat was added to an aqueous solution comprising meglumine and cyclodextrin at a different ratio to form a suspension. The suspension was placed in a constant-temperature incubation shaker, and was shaken at a temperature of 25±1° C., 37±1° C. and 60±1° C. for 72 h, respectively. After reaching equilibrium, the suspension was filtered through a 0.45 μm hydrophilic filter membrane to obtain a filtrate. After the filtrate was properly diluted with an acetonitrile-water (50:50, v/v) solution, the absorbance was measured at 224 nm, and the solubility of Tecovirimat was calculated. The corresponding solubility curve was plotted by using the solubility of the drug as the ordinate, and using the concentration of cyclodextrin and/or meglumine as the abscissa. The solubility curves of Tecovirimat at 25° C., 37° C. and 60° C. were shown in
As seen from
As seen from
As seen from
According to the experimental result of solubility, when the amount of cyclodextrin was in a range from 5% to 40% (w/v), the thermodynamic parameters were calculated for the inclusion complex formed by the drug and cyclodextrin. The result was shown in Table 5. A negative ΔG indicated that inclusion process could occur spontaneously; a positive ΔH indicated that the main driving force of inclusion process was hydrophobic interaction, and meanwhile the inclusion process was an endothermic reaction, and the inclusion reaction could be promoted by increasing the temperature properly. However, the ternary composition had a lower ΔH value because the entrance of meglumine into the cavity resulted in the release of more enthalpy-rich water. A positive ΔS indicated that the inclusion process was an enthalpy increasing process, and the ternary composition had a lower ΔS value because the degree of freedom for Tecovirimat and meglumine in the cavity was reduced, and a more stable inclusion system was formed.
Experimental method: a suitable amount of Tecovirimat, meglumine, cyclodextrin, a binary composition (Tecovirimat/meglumine at a weight ratio of 1:2), a binary composition of (Tecovirimat/cyclodextrin at a weight ratio of 1:6), and a ternary composition (Tecovirimat/meglumine/cyclodextrin at a weight ratio of 1:2:6) were dissolved in DMSO-d6 to prepare samples, respectively, and the possible intermolecular interaction was analyzed by 1H NMR spectroscopy.
Experimental result: the NMR spectra were shown in
To sum up, as compared with the prior art, the present invention provides a pharmaceutical composition of Tecovirimat; the addition of an additive makes the drug to be included more easily, which greatly enhance the inclusion efficiency of cyclodextrin and simplifies the operation; and meanwhile, the amount of cyclodextrin used is reduced, thereby reducing the potential medicament risk. The pharmaceutical composition also has the advantages such as simple formulation, low cost, easy operation, stable and controllable quality, and good reproducibility.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201610559479.1 | Jul 2016 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2017/091650 | 7/4/2017 | WO | 00 |
