Tecovirimat composition, preparation method therefor, and use thereof

Abstract

Disclosed are a tecovirimat composition, a preparation method therefor, and use thereof. The composition comprises an active ingredient tecovirimat, an inclusion agent cyclodextrin, a solubilizer meglumine, and a basic additive in a weight ratio of 1:(15-30):(0.05-0.2):(0.1-0.2). The use of a combination of the three excipients, i.e., the inclusion agent, the solubilizer, and the basic additive, in a ratio can improve the solubility of the active ingredient tecovirimat. Compared with the prior art, the amount of each excipient used in the composition can meet the prescribed limit in relevant regulations, the risks of allergies, hemolysis, and irritation are all relatively low, and the safety of the formulation is better. In addition, the method provided for preparing the composition is simple and easy to implement, the quality of the finished formulation is more stable, and the application is more convenient. The composition can be used for treating or preventing viral infections and related diseases caused by orthopoxviruses.

Description

A tevilima composition, its preparation method and application Technical Field

[0001] This invention relates to the pharmaceutical field, and more specifically, to a tevirima composition, its preparation method, and its application. Background Technology

[0002] Tecovirimat (English name: Tecovirimat; research code: ST-246; formerly known as Tecovirim; Formula 1), chemically named N-[(3aR,4R,4aR,5aS,6S,6aS)-3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-vinylcyclopropyl[f]isoindol-2(1H)-yl]-4-(trifluoromethyl)benzamide monohydrate, is an effective broad-spectrum inhibitor of orthopox virus infection, including smallpox, monkeypox, and cowpox. This compound targets the conserved protein VP37 of orthopox virus, preventing the formation of virus-specific encapsulated complexes, thereby inhibiting the production of infectious enveloped viral particles and blocking intercellular and long-distance viral transmission. However, tecovirimat has a water solubility of approximately 3.35 μg / mL, classifying it as a nearly insoluble or insoluble compound, a characteristic that severely limits its clinical application.

[0003] Chinese patent CN103281898 discloses a formulation for tevirimal injection, which uses hydroxypropyl-β-cyclodextrin as an excipient to improve the solubility of tevirimal (up to 21.23 mg / mL). Based on this patent, a tevirimal injection (marketed under the name: [brand name missing]) has been developed. Hereinafter referred to as binary prescriptions, binary compositions, The dosage of hydroxypropyl-β-cyclodextrin in injectable formulations is as high as 40%, far exceeding the regulatory limits according to clinical usage protocols. Studies have shown that high doses of hydroxypropyl-β-cyclodextrin pose a risk of hemolysis and kidney-related toxicity. Injectable tevirima is contraindicated in patients with severe renal impairment. It is worth noting that tevirima has been approved for monkeypox treatment; however, a large proportion of monkeypox patients are children with underdeveloped renal function or severe renal impairment, thus significantly limiting its use. Clinical applications. The injection is a solution and must be stored and transported at 2-8℃.

[0004] Patent CN107625967 discloses another injectable formulation of tevirimal (hereinafter referred to as a ternary composition), which uses cyclodextrin and meglumine as excipients. The amount of meglumine used (calculated based on the dosing regimen of similar marketed injectables) far exceeds the 75 mg / day limit specified in the US Excipient Database (IID). Furthermore, due to insufficient safety data regarding excessive use of meglumine as an excipient, it cannot be directly used for commercial development. In addition, this patented formulation requires preparation at high temperature (60°C), placing high demands on preparation conditions.

[0005] Therefore, developing a new formulation that is safer, more stable, and suitable for commercial development is an urgent problem to be solved. Summary of the Invention

[0006] One object of this invention is to provide a tevirimal composition and its key formulation for the treatment of vaccinia virus infection. To address the problems existing in current tevirimal injections, this invention explores novel formulations. Cyclodextrin is used in the solubilization of various poorly soluble drugs, but high doses pose safety risks such as nephrotoxicity and hemolysis; therefore, the dosage of cyclodextrin must be controlled within an appropriate range. Furthermore, tevirimal is a weakly acidic compound, and alkaline conditions can promote its dissolution. However, tevirimal contains an amide structure, and excessive use of alkali can easily lead to the degradation of the compound, increasing impurity content and affecting the stability of the formulation. This invention investigated various excipients and their dosages, discovering a composition that can significantly improve the solubility of tevirimal. Unexpectedly, it was found that the simultaneous use of the inclusion agent cyclodextrin, the solubilizer meglumine, and the alkaline additive (referred to in this invention as a quaternary formulation or quaternary composition) is necessary to completely dissolve the active ingredient tevirimal while maintaining good stability, provided that the dosages of each excipient comply with relevant regulatory requirements. Using one or two of the inclusion agent cyclodextrin, the solubilizer meglumine, and the basic additive as excipients can lead to problems such as incomplete dissolution of tevirimal or excipient safety risks. Therefore, the proper combination of these three excipients—cyclodextrin, meglumine, and basic additives—is crucial to ensuring the complete dissolution of the active ingredient tevirimal and the safety of the formulation.

[0007] The tevirima quaternary composition provided by this invention meets regulatory requirements in terms of the amount of each excipient, and the safety of the excipients is fully guaranteed. While improving the solubility of the active ingredient tevirima, it also enhances the stability of the formulation. The formulation has low risks of allergy, hemolysis and irritation, and high safety, making it suitable for commercial development.

[0008] Another object of the present invention is to provide a method for preparing the tevirima composition.

[0009] Another object of the present invention is to provide the use of the tevirima composition.

[0010] To achieve the above objectives, in one respect, the present invention provides a tevirimal composition, wherein the composition contains at least tevirimal, an inclusion agent, a solubilizer, and an alkaline additive, in a weight ratio of 1:(15-30):(0.05-0.2):(0.1-0.2);

[0011] The inclusion agent is selected from one or more combinations of β-cyclodextrin, methyl-β-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, 3-hydroxypropyl-β-cyclodextrin, 2,3-dihydroxypropyl-β-cyclodextrin or sodium sulfobutyl-β-cyclodextrin;

[0012] The solubilizer is selected from one or more combinations of meglumine, hydroxypropyl methylcellulose and copovidone;

[0013] The alkaline additive is selected from one or more of organic or inorganic bases and their combinations.

[0014] According to some specific embodiments of the present invention, the weight ratio of tevitroma, inclusion agent, solubilizer and alkaline additive is 1:(15-20):(0.05-0.15):(0.1-0.15).

[0015] According to some specific embodiments of the present invention, the weight ratio of tevirima, inclusion agent, solubilizer and alkaline additive is 1:15:0.1:0.1.

[0016] According to some specific embodiments of the present invention, the inorganic base is selected from hydroxides, carbonates or bicarbonates of alkali metals or alkaline earth metals; the organic base is selected from triethanolamine or arginine.

[0017] According to some specific embodiments of the present invention, the inorganic base is selected from sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, or potassium bicarbonate.

[0018] According to some specific embodiments of the present invention, wherein,

[0019] The inclusion agent is one or a combination of 2-hydroxypropyl-β-cyclodextrin, 3-hydroxypropyl-β-cyclodextrin and 2,3-dihydroxypropyl-β-cyclodextrin; preferably one or a combination of two of 2-hydroxypropyl-β-cyclodextrin and 3-hydroxypropyl-β-cyclodextrin; more preferably 2-hydroxypropyl-β-cyclodextrin.

[0020] The solubilizer is one or more of meglumine, hydroxypropyl methylcellulose, and copovidone; preferably meglumine.

[0021] The alkaline additive is one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, triethanolamine, and arginine; preferably sodium hydroxide or potassium hydroxide; more preferably sodium hydroxide.

[0022] According to some specific embodiments of the present invention, the inclusion agent is selected from one or more combinations of 2-hydroxypropyl-β-cyclodextrin, 3-hydroxypropyl-β-cyclodextrin and 2,3-dihydroxypropyl-β-cyclodextrin;

[0023] The solubilizer is meglumine;

[0024] The alkaline additive is sodium hydroxide.

[0025] According to some specific embodiments of the present invention, the inclusion agent is selected from one or a combination of two of 2-hydroxypropyl-β-cyclodextrin and 3-hydroxypropyl-β-cyclodextrin.

[0026] According to some specific embodiments of the present invention, the inclusion agent is 2-hydroxypropyl-β-cyclodextrin.

[0027] According to some specific embodiments of the present invention, the tevirimal composition is tevirimal powder for injection or tevirimal injection solution.

[0028] According to some specific embodiments of the present invention, when the teviquilmab composition is an injection solution, the concentration of teviquilmab is 5-50 mg / mL; preferably 10-20 mg / mL;

[0029] According to some specific embodiments of the present invention, when the tevirima composition is an injection solution, the pH value of the injection solution is 8-12.

[0030] According to some specific embodiments of the present invention, when the tevirima composition is an injection solution, the pH value of the injection solution is 9-11.

[0031] According to some specific embodiments of the present invention, when the tevirimal composition is a powder for injection, after reconstitution at a tevirimal concentration of 5-50 mg / mL, the pH value of the resulting solution is 8-12.

[0032] According to some specific embodiments of the present invention, when the tevirimal composition is a powder for injection, it can be reconstituted at a tevirimal concentration of 10-20 mg / mL before use.

[0033] According to some specific embodiments of the present invention, when the tevirima composition is a powder for injection, the pH value of the solution obtained after reconstitution is 9-11.

[0034] According to some specific embodiments of the present invention, when the tevirimal composition is a powder for injection, after reconstitution at a tevirimal concentration of 10-20 mg / mL, the pH value of the resulting solution is 9-11.

[0035] According to some specific embodiments of the present invention, when the tevirima composition is a powder for injection, its reconstitution solvent is one or more of water for injection, 0.9% sodium chloride and 5% glucose solution.

[0036] On the other hand, the present invention also provides a method for preparing a teviquirida composition, comprising the following steps:

[0037] (1) Take an appropriate amount of water for injection, add an inclusion agent, a solubilizer and an alkaline additive, stir and dissolve to obtain the first solution;

[0038] (2) Add tevirimal to the first solution, add the remaining water for injection, and stir until the solid dissolves to obtain tevirimal injection solution; or

[0039] Optionally, step (3) is also included: freeze-drying or spray-drying the injection solution obtained in step (2) to obtain tevirima powder for injection.

[0040] According to some specific embodiments of the present invention, step (1) includes taking 60%-80% of the total amount of water for injection, adding an inclusion agent, a solubilizer and an alkaline additive, and stirring to dissolve to obtain a first solution.

[0041] According to some specific embodiments of the present invention, step (1) involves controlling the temperature of water for injection to be no higher than 60°C and adding an inclusion agent, a solubilizer, and an alkaline additive.

[0042] According to some specific embodiments of the present invention, step (1) involves controlling the temperature of water for injection to be not lower than 15°C and adding an inclusion agent, a solubilizer and an alkaline additive.

[0043] According to some specific embodiments of the present invention, step (1) involves controlling the temperature of water for injection at 15°C-60°C and adding an inclusion agent, a solubilizer, and an alkaline additive.

[0044] According to some specific embodiments of the present invention, step (2) includes adding tevirima to the first solution and adding the remaining water for injection, and sterilizing to obtain tevirima injection solution.

[0045] According to some specific embodiments of the present invention, the sterilization in step (2) is either filtration sterilization or autoclaving. The filtration sterilization filter membrane has a diameter of 0.22 μm to 0.45 μm; the autoclaving parameters are 115℃ to 121℃ for 15 min to 30 min.

[0046] According to some specific embodiments of the present invention, step (2) involves controlling the temperature of the first solution to be no higher than 60°C and adding tevirima.

[0047] According to some specific embodiments of the present invention, step (2) involves controlling the temperature of the first solution to be not lower than 15°C and adding tevirima.

[0048] According to some specific embodiments of the present invention, step (2) involves controlling the temperature of the first solution at 15°C-60°C and adding tevirima.

[0049] The tevirima injection obtained by this invention can be used directly in combination with other solutions; while the tevirima powder injection obtained by this invention can be reconstituted and used with water for injection, 0.9% sodium chloride, or 5% glucose solution.

[0050] In another aspect, the present invention also provides the use of the tevirima composition in the preparation of medicaments for the treatment or prevention of viral infections.

[0051] According to some specific embodiments of the present invention, the virus is orthopoxvirus.

[0052] According to some specific embodiments of the present invention, the virus is vaccinia virus, monkeypox virus, smallpox virus, or vaccinia virus.

[0053] It is understood that, without contradiction, the various specific embodiments of the present invention can be combined arbitrarily.

[0054] The term "optional" in this invention means that the condition or situation may occur or exist, or may not occur or not exist. For example, the term "optionally includes step (3)" in this invention means that step (3) may or may not exist. When step (3) exists, tevirimal powder injection is obtained, and when step (3) does not exist, tevirimal injection solution is obtained.

[0055] In summary, this invention provides a tevirimal composition, its preparation method, and its application. The tevirimal composition of this invention has the following advantages:

[0056] 1) Compared to existing technologies, this invention develops a quaternary formulation of tevirimal and determines its key proportions. This formulation improves the solubility of tevirimal while ensuring that the dosage of each excipient meets the requirements of relevant regulations, making it suitable for commercial development.

[0057] 2) Compared with the patent CN107625967, the preparation method of the tevirima quaternary formulation of the present invention is simple, with low impurity content, which ensures the repeatability and quality stability of the product.

[0058] 3) The quaternary composition provided by this invention has a lower risk of allergy, hemolysis, and irritation. Compared to already marketed... Injectable solutions offer better safety and can cover a wider range of people (such as patients with severe kidney damage).

[0059] 4) The quaternary composition provided by this invention and Compared to the injectable form, it significantly improves the solubility of tevimet, has a higher temperature tolerance, and better stability, allowing for transportation, storage, and use at room temperature, greatly improving the convenience of clinical application.

[0060] This invention provides a novel tevirivima composition with a simple preparation method, higher safety, and better stability, as well as a method for its preparation, suitable for commercial development. This composition can be used to treat or prevent viral infections and related diseases caused by orthopoxvirus. Attached Figure Description

[0061] Figure 1 shows the results of an in vitro hemolysis comparison study between the tevirima quaternary composition and the binary composition in Example 13. Detailed Implementation

[0062] The following detailed embodiments illustrate the implementation process and beneficial effects of the present invention, aiming to help readers better understand the essence and characteristics of the present invention, and are not intended to limit the scope of implementation of this case.

[0063] Example 1: Formulation of the teviquirida composition

[0064] Preparation method: As shown in Table 1, weigh the corresponding weights of hydroxypropyl-β-cyclodextrin, meglumine and sodium hydroxide in each formulation, dissolve them in a certain volume of water for injection at room temperature, stir and mix evenly, then weigh 1g of tevirima and add it to the solution, add water for injection to 100mL, stir at room temperature, and observe the dissolution after 24 hours.

[0065] Table 1. Solubility of Tivirima compositions with different ratios

[0066] Experimental Results: According to the relevant regulations of the US FDA and the EU regarding pharmaceutical excipients, the maximum dosage of each excipient is as follows: hydroxypropyl-β-cyclodextrin 200 mg / kg / day, meglumine 75 mg / day, and sodium hydroxide 78 mg / day. Based on the dosing regimens of similar marketed formulations (see Table 6), for individuals of normal weight, under the above dosage limits, it was found that tevirimal could only be completely dissolved when all three excipients were used simultaneously; using any two of the three excipients failed to achieve complete dissolution. Therefore, the optimal combination of the inclusion compound cyclodextrin, the solubilizer meglumine, and the alkaline additive sodium hydroxide is crucial to ensuring the complete dissolution of the active ingredient tevirimal and the safety of the formulation.

[0067] Example 2: Solubility of the tevirima quaternary composition

[0068] Preparation method: As shown in Table 2, weigh the corresponding weights of hydroxypropyl-β-cyclodextrin, meglumine and sodium hydroxide in each formulation, dissolve them in a certain volume of water for injection at room temperature, stir and mix evenly, then weigh 2-5g of tevirima and add it to the solution, adjust the pH and add water for injection to 100mL, continue stirring, and observe the dissolution after 24 hours.

[0069] Table 2 Solubility of Teverimilma Quaternary Compositions

[0070] Experimental Results: All the above formulations can completely dissolve tevirime, and the formulation provided by this invention can increase the solubility of tevirime to 50 mg / mL, which is beneficial to the industrial production of tevirime preparations. During the experiment, it was also observed that: as the amount of tevirime increases, the time required for the formulation to dissolve becomes longer; when the tevirime content is limited, the more excipients in the formulation, the shorter the time required for complete dissolution.

[0071] Example 3: Comparison of preparation conditions between tevirima quaternary and ternary compositions

[0072] The tevirima quaternary composition was prepared according to the preparation method in Example 2, and the ternary composition was prepared according to the preparation method in patent CN107625967. The preparation volume was 100 mL. Each formulation was stirred at a specific temperature. After 24 hours, the dissolution state of each composition was recorded. The results are shown in Table 3.

[0073] Table 3. Comparison of the solubility states of each formulation of the tevirima quaternary composition and the ternary composition. Note: The formulation ratios of the ternary compositions are taken from the preferred formulation of patent CN107625967. Ternary composition 1 is tevirime: meglumine: hydroxypropyl-β-cyclodextrin = 2g: 4g: 12g, and ternary composition 2 is tevirime: meglumine: hydroxypropyl-β-cyclodextrin = 5g: 10g: 30g.

[0074] Experimental results: All formulations of the tevirima quaternary composition were completely soluble at preparation temperatures ranging from 25°C to 60°C, while the ternary composition prepared according to the patent formulation of CN107625967 required a preparation temperature of 60°C to be completely soluble.

[0075] Example 4: Determination of impurities in tivalima quaternary composition solution

[0076] The solution prepared in Example 2 was left at room temperature for 12 hours, and its appearance, pH and the content of related substances were measured. The results are shown in Table 4.

[0077] Table 4. Solution stability of quaternary compositions

[0078] Experimental results: After 12 hours of storage, the appearance and pH of each formulation of the tevirima quaternary composition did not change, and the impurity content was at a low level, indicating that the quaternary composition solution has good stability.

[0079] Example 5: Reconstitution and compatibility stability study of tivalima quaternary composition

[0080] The quaternary composition solution was prepared according to the method in Example 2, dispensed (10 mL / vial), and freeze-dried to obtain a powder for injection. The powder was reconstituted with 20 mL of water for injection, and 40 mL of 5% glucose or 0.9% sodium chloride was added to obtain a compatible solution. The prepared solution was left at room temperature for 6 hours, and the pH and the content of related substances were measured. The results are shown in Table 5.

[0081] Table 5. Four-component freeze-dried powder * Compatibility stability Note: *The freeze-drying process here is as follows: (1) Pre-freezing and heat preservation: pre-freezing and heat preservation at -40℃; (2) Vacuuming; (3) When the vacuum degree is lower than 0.15mbar, start heating: increase the temperature from -20℃ to 35℃.

[0082] Experimental results: After reconstitution and compatibility at room temperature and standing for 6 hours, the lyophilized powder of the tevirima quaternary composition showed no significant changes in any of the measured items, indicating that the lyophilization process of each formulation of the quaternary composition was stable and the compatibility was good.

[0083] Example 6: Comparison of excipient dosage between quaternary composition formulation 10 and existing patented formulations at the intended clinical dosage

[0084] according to Injection instructions, calculate prescription 10, Dosage of excipients for injection solutions, patented formulations (CN103281898), and ternary compositions in patients with different body weights (see Tables 6 to 8).

[0085] Table 6 Comparison of hydroxypropyl-β-cyclodextrin excipient dosage for different formulations Note: The EU Annex to the Guidelines on Excipients in Labelling and Packaging Instructions for Human Use of Pharmaceutical Products stipulates that the limit for hydroxypropyl-β-cyclodextrin in injectable formulations is 200 mg / kg / day. The US Excipient Database (IID) lists the limit for hydroxypropyl-β-cyclodextrin in injectable formulations as 9600 mg / day. (Prescription a is from...) Injection solution instructions. b is from patent CN103281898, and is the formulation with the lowest amount of hydroxypropyl-β-cyclodextrin in all embodiments of the patent.

[0086] Table 7 Comparison of excipient dosage for meglumine in different formulations Note: According to the U.S. Excipients Database (IID), the limit for meglumine excipients for injection (including lyophilized powder) is 75 mg / day. a The ternary composition formulation is extracted from patent CN107625967.

[0087] Table 8. Dosage of NaOH excipient in quaternary composition formulation 10 Note: According to the U.S. Ingredients Database (IID), the limit for NaOH in injections is 78 mg / day.

[0088] Results: The amounts of each excipient in the teviquilimab quaternary compound formulation 10 met the corresponding excipient limits at clinical dosages. The dosage of hydroxypropyl-β-cyclodextrin in the injectable formulation and patent CN103281898 far exceeds the limits specified in the EU's Annex to the Guidelines on Excipients in the Labelling and Packaging Instructions for Human Use of Pharmaceutical Products; while in the ternary composition from patent CN107625967, the dosage of meglumine, an excipient, far exceeds the limits specified in the US Excipient Database (IID). These results indicate that the tevirima quaternary composition has better excipient safety and is suitable for commercial development.

[0089] Example 7: Comparative Study on the Compatibility Stability of Different Prescription Formulations

[0090] Following the method in Example 5, a lyophilized powder of the quaternary composition formulation 10 was prepared and reconstituted using 0.9% sodium chloride injection or 5% glucose injection. (Reference) The binary composition of the injection solution was prepared, with each vial containing 20 mL. All the above-mentioned different formulations were diluted with twice the volume of 0.9% sodium chloride injection or 5% glucose injection. The diluted solutions were placed at 2-8℃, and various indicators were measured. The results are shown in Table 9.

[0091] Table 9 Comparative Study of Compatibility Stability of Different Formula Preparations Note: The ratio of the binary formulation is tevimet: hydroxypropyl-β-cyclodextrin = 1:40, stirred until dissolved.

[0092] Experimental Results: When the tevirima tetravalent formulation (Formula 10) and the binary formulation were mixed with 0.9% sodium chloride or 5% glucose, respectively, and incubated at 2-8℃ for 168 hours, no significant changes were observed in any of the test items for formulation 10. However, the binary formulation injection showed turbidity after 48 hours of mixing with either 0.9% sodium chloride or 5% glucose. These results indicate that formulation 10 has better compatibility stability than the binary formulation.

[0093] Example 8: High-temperature stability study of different formulations

[0094] Following the method in Example 5, a quaternary composition lyophilized powder (Formula 10) was prepared. It was then placed at 60°C for 10 days or at 40°C for 30 days, reconstituted with 20 mL of water for injection, and various indicators were measured. Binary compositions and ternary compositions 1 were prepared according to Examples 7 and 3, respectively, and placed at 60°C for 10 days or at 40°C for 30 days, and various indicators were measured. The results are shown in Table 10.

[0095] Table 10 High-temperature stability of different formulations

[0096] Experimental results: After preparation, the initial impurity level of ternary composition 1 was higher than that of formulation 10 and binary composition. After each formulation was placed at 60°C for 10 days or at 40°C for 30 days, the impurity growth trend in binary composition and ternary composition 1 was significant, far exceeding the impurity level of quaternary composition formulation 10.

[0097] Example 9: Study on the high humidity stability of quaternary compositions

[0098] Following the method in Example 5, the quaternary composition lyophilized powder was prepared using Formula 10. It was placed at RH 92.5% for 30 days and reconstituted with 20 mL of water for injection on day 0, day 10, and day 30, respectively. Various indicators were measured, and the results are shown in Table 11.

[0099] Table 11. Stability test results of Formulation 10 under high humidity conditions.

[0100] Experimental results: After 30 days of storage at RH 92.5%, the tevirima tetrapolymer formulation 10 showed no significant changes in any of the measured parameters, indicating that it has good stability under high humidity conditions.

[0101] Example 10: Study on the photostability of tivalima quaternary compositions

[0102] Following the method in Example 5, the quaternary composition lyophilized powder was prepared using Formula 10. It was placed at 4500±500 Lx for 30 days and reconstituted with 20 mL of water for injection on day 0, day 10, and day 30, respectively. Various indicators were measured, and the results are shown in Table 12.

[0103] Table 12 Results of stability tests on formulation 10 under light conditions.

[0104] Experimental results: After 30 days of storage at 4500±500 Lx, the tevirima tetrapolymer formulation 10 showed no significant changes in any of the measured parameters, indicating good stability under light conditions.

[0105] Example 11: Stability study of tivalima quaternary composition at 40°C and 75% RH

[0106] Following the method in Example 5, the quaternary composition lyophilized powder was prepared using Formula 10. It was placed at 40°C and RH 75% for 90 days, and then reconstituted with 20 mL of water for injection on day 0, day 30, day 60, and day 90, respectively. Various indicators were measured, and the results are shown in Table 13.

[0107] Table 13 shows the stability test results of Formulation 10 at 40°C and 75% RH.

[0108] Experimental results: After 90 days of storage at 40℃ and 75% RH, the tevirima tetrapolymer formulation 10 showed no significant changes in any of the measured parameters, indicating that it has good stability under these conditions.

[0109] Example 12: Stability study of tivalima quaternary composition at 25°C and 60% RH

[0110] Following the method in Example 5, the quaternary composition lyophilized powder was prepared using Formula 10. It was placed at 25°C and RH 60% for 90 days, and then reconstituted with 20 mL of water for injection on day 0, day 30, day 60, and day 90, respectively. Various indicators were measured, and the results are shown in Table 14.

[0111] Table 14 shows the stability test results of Formulation 10 at 25°C and 60% RH.

[0112] Experimental results: After 90 days of storage at 25℃ and 60% RH, the tevirima tetrapolymer formulation 10 showed no significant changes in any of the measured parameters, indicating that it has good stability under these conditions.

[0113] Example 13: Comparative Study of Hemolytic Risk in Different Prescription Formulations

[0114] Following the method in Example 5, a quaternary composition and a corresponding excipient control group (excluding tivarimal) were prepared using Formula 10. A binary composition and a corresponding excipient control group (excluding tivarimal) were prepared according to Example 7. The above four compositions were then formulated using the same method. The corresponding amounts of the formulated solutions were added to the in vitro hemolysis test system of New Zealand rabbit erythrocytes, and the mixture was incubated at 36.1°C for 3 hours. The hemolysis was recorded, and the results are shown in Table 15 and Figure 1.

[0115] Table 15 Results of hemolysis experiments for different formulations

[0116] Experimental results: Hemolysis was observed in two high-dose tubes of the binary composition and its excipient control group, while no hemolysis was observed in the quaternary composition and its excipient control group. This indicates that the quaternary composition formulation 10 has no risk of hemolysis and its hemolysis safety is higher than that of the binary composition.

[0117] Example 14: Allergy Risk Study of Tevilima Quadrivalent Combination

[0118] Following the method described in Example 5, a quaternary composition formulation was prepared using Formula 10. It was then prepared with 0.9% sodium chloride injection and set aside for use. An active sensitization experiment was conducted in a guinea pig model, using 0.9% sodium chloride injection as a negative control and albumin as a positive control. All groups of animals underwent intraperitoneal injection for sensitization and intravenous injection for challenge. Challenge was performed twice, on days 14 and 21, following the last sensitization injection. During this period, cage-side observation, detailed clinical observation, weight recording, post-challenge observation, and allergy evaluation were conducted.

[0119] Experimental results: All animals in the positive control group died after challenge, while the negative control group and the formulation 10 group showed no abnormalities, indicating that the tevirima tetravalent combination formulation 10 has no risk of allergic reaction.

[0120] Example 15: Risk Study of Vascular Irritation in Rabbits Using Teveremil Quadrivalent Composition

[0121] Following the method described in Example 5, a quaternary composition formulation was prepared using Formula 10. It was then prepared with 0.9% sodium chloride solution and set aside for use. Vascular stimulation studies were conducted in a New Zealand white rabbit model. The administration route was twice daily, with each infusion lasting 60 minutes, for 7 consecutive days, followed by a 14-day recovery period. During this period, cage-side observation, detailed clinical observation, administration site observation, weight recording, and post-dissection evaluation were performed. The experimental results are shown in Table 16.

[0122] Table 16 Results of rabbit vascular stimulation study of teverlima tetravalent composition

[0123] Experimental results: Formulation 10 of the tevirima tetrapolymer combination showed no risk of irritation.

[0124] Example 16: Pharmacokinetic Analysis of the Tevirimal Quadrivalent Combination in Cynomolgus Monkeys

[0125] The quaternary composition (Formula 10) was prepared according to the method in Example 5 and prepared with 0.9% sodium chloride solution. It was administered intravenously to cynomolgus monkeys at a dose of 10 mg / kg over 4 hours. Blood samples were collected at 14 points before and after administration to determine the plasma concentration of teviriva at each time point and to calculate the main pharmacokinetic parameters. The results are shown in Table 17.

[0126] Table 17 Pharmacokinetic parameters of the quaternary composition in cynomolgus monkeys

[0127] Experimental results showed that after intravenous infusion of the quaternary combination, the plasma concentration of teviquirimal reached its peak at the end of the infusion, corresponding to C0.05. max AUC (0-∞) The values ​​were 5048 ng / mL and 23672 h·ng / mL, respectively.

[0128] Example 17: Evaluation Experiment of the Efficacy of Teverimilar against Orthozoma Virus

[0129] Serially diluted tevirima was co-incubated with Vero cells and orthopoxviruses (such as vaccinia virus, vaccinia virus, and monkeypox virus) for 3-5 days. The anti-orthopoxvirus activity of tevirima was detected by the cytopathic effect (CIE) assay. 50 (Half-maximal inhibitory concentration). The cytotoxicity of teviriva was detected using a similar system (without virus) (CC). 50 (Cellular half-maximal toxicity concentration). The experimental results are shown in Table 18.

[0130] Table 18. Anti-orchia virus activity and cytotoxicity of teverlimab

[0131] Experimental results: Teverima exhibits good antiviral activity against orthopoxviruses (vaccinia virus, cowpox virus, monkeypox virus), and no cytotoxicity was detected in the system.

[0132] In summary, this invention provides a quaternary composition of tevirimal, its preparation method, and its application. The formulation includes the active ingredient tevirimal, the inclusion agent cyclodextrin, the solubilizer meglumine, and an alkaline additive. The weight ratio of each component is tevirimal: cyclodextrin: meglumine: alkaline additive = 1:(15-30):(0.05-0.2):(0.1-0.2). It is worth noting that the combination of the three excipients is crucial to ensuring the complete dissolution of the active ingredient tevirimal and the safety of the formulation. Compared to existing technologies, the preferred quaternary composition preparation method of this invention is simple. While improving the solubility of tevirimal, the dosage of each excipient meets relevant regulatory requirements. Combined with the results of allergy, hemolysis, and irritation tests, this quaternary composition exhibits good formulation safety. Multiple stability tests, including those on influencing factors (high temperature, high humidity, light), reconstitution / compatibility, accelerated reaction, and long-term effects, show that this quaternary composition has good stability and can be stored, transported, and used at room temperature. Injectable formulations are more convenient than oral administration. Pharmacological studies have shown that tevirimal possesses broad-spectrum anti-orchioplasmosis activity and is non-cytotoxic. Therefore, this invention provides a novel tevirimal quaternary composition with enhanced safety, improved stability, and ease of large-scale preparation, suitable for commercial development. This composition can be used to treat or prevent viral infections and related diseases caused by orchioplasmosis.

Claims

1. A tevirimal composition, said composition comprising at least tevirimal, an inclusion agent, a solubilizer, and an alkaline additive, in a weight ratio of 1:(15-30):(0.05-0.2):(0.1-0.2); The inclusion agent is selected from one or more combinations of β-cyclodextrin, methyl-β-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, 3-hydroxypropyl-β-cyclodextrin, 2,3-dihydroxypropyl-β-cyclodextrin or sodium sulfobutyl-β-cyclodextrin; The solubilizer is selected from one or more combinations of meglumine, hydroxypropyl methylcellulose and copovidone; The alkaline additive is selected from one or more of organic or inorganic bases and their combinations.

2. The teviquirida composition according to claim 1, wherein, The inclusion agent is one or a combination of 2-hydroxypropyl-β-cyclodextrin, 3-hydroxypropyl-β-cyclodextrin and 2,3-dihydroxypropyl-β-cyclodextrin; preferably one or a combination of two of 2-hydroxypropyl-β-cyclodextrin and 3-hydroxypropyl-β-cyclodextrin; more preferably 2-hydroxypropyl-β-cyclodextrin. The solubilizer is one or more of meglumine, hydroxypropyl methylcellulose, and copovidone; preferably meglumine. The alkaline additive is one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, triethanolamine, and arginine; preferably sodium hydroxide or potassium hydroxide; more preferably sodium hydroxide.

3. The tevilima composition according to claim 1 or 2, wherein: The weight ratio of tevirima, inclusion agent, solubilizer and alkaline additive is 1:(15-20):(0.05-0.15):(0.1-0.15); preferably 1:15:0.1:0.

1.

4. The tevilima composition according to claim 1, wherein the dosage form of the composition is a powder for injection or an injection solution.

5. The tevirimal composition according to claim 4, wherein when the tevirimal composition is an injection solution, the concentration of tevirimal is 5-50 mg / mL; preferably 10-20 mg / mL.

6. The tevirimal composition according to claim 4, wherein when the tevirimal composition is an injection solution, the pH value of the injection solution is 8-12 (preferably 9-11); and when the tevirimal composition is a powder for injection, after reconstitution at a tevirimal concentration of 5-50 mg / mL (preferably 10-20 mg / mL), the pH value of the resulting solution is 8-12 (preferably 9-11).

7. The tevirimal composition according to claim 4 or 6, wherein when the tevirimal composition is a powder for injection, its reconstitution solvent or compatibility solvent is one or more of water for injection, 0.9% sodium chloride and 5% glucose solution.

8. A method for preparing the teviquiridar composition according to any one of claims 1 to 7, wherein, Includes the following steps: (1) Take an appropriate amount of water for injection, add an inclusion agent, a solubilizer and an alkaline additive, stir and dissolve to obtain the first solution; (2) Add tevirimal to the first solution, add the remaining water for injection, and stir until the solid dissolves to obtain tevirimal injection solution; or Optionally, step (3) is also included: freeze-drying or spray-drying the injection solution obtained in step (2) to obtain tevirima powder for injection.

9. The preparation method according to claim 8, wherein, Step (2) involves adding tevirima to the first solution and adding the remaining water for injection, followed by sterilization (preferably aseptic filtration or autoclaving) to obtain tevirima injection.

10. The use of the tevirima composition according to any one of claims 1 to 7 in the preparation of a medicament for treating or preventing viral infections.

11. The application according to claim 10, wherein, The virus is orthopoxvirus; preferably, it is vaccinia virus, monkeypox virus, smallpox virus, or vaccinia virus.

Images