Freeze-dried hydroxyurea methyl acyl fulvenes

Abstract

The lyophilized formulations comprise steepstercetin, including hydroxyurea methyl acyl fulvene and pharmaceutically acceptable salts thereof, in combination with a cyclodextrin. These formulations are designed to enhance the solubility, stability and therapeutic efficacy of steep steep cephalosporin, which has been illustrated due to its potent anti-cancer properties. Methods for producing these lyophilized formulations are provided, including a lyophilization process.

Description

Technical Field

[0001] This disclosure relates to lyophilized formulations or compositions of illudin (e.g., illudin analogs) and methods for preparing and using them, for example, as cancer therapeutic agents. Background Technology

[0002] The use of lyophilized (freeze-dried) drugs may be beneficial in cancer treatment or management. Lyophilization maintains purity, extends shelf life, and stabilizes a variety of pharmaceutical products, including those that are relatively unstable in solution or contain heat-labile active ingredients, as well as vaccines. Lyophilized therapeutic agents are easier to reconstitute in the pharmacy before patient administration. Careful selection of excipients for lyophilization, based on the properties of the active pharmaceutical ingredient, allows lipophilic active molecules to be reconstituted without loss of concentration due to potential precipitation. Lyophilization has also been shown to be effective in enhancing drug stability.

[0003] Freeze-drying is widely used across various industries. In the pharmaceutical field, it protects sensitive drugs, vaccines, and diagnostic reagents. In the food industry, it preserves fruits, vegetables, instant coffee, and herbs. It is also used to preserve historical documents and biological samples. Freeze-drying extends the shelf life of biological materials, food, and pharmaceuticals by freezing materials and removing frozen components through sublimation, thereby eliminating moisture and maintaining structural integrity.

[0004] When formulated in organic media or mixtures of organic and aqueous media, cryptocephalin family compounds are found to be unstable during storage and transport at room temperature. Cryptocephalins and their analogues (e.g., hydroxyurea methyl acyl fulene) face challenges in lyophilization due to their inherent chemical properties. Cryptocephalins exhibit limited solubility in water, making it difficult to formulate aqueous solutions for lyophilization, as they tend to precipitate or form unstable suspensions. Furthermore, cryptocephalins can degrade under certain conditions such as heat, light, or pH changes. The complex chemical structure of cryptocephalins, including hydroxyurea methyl acyl fulene with multiple hydroxyl groups and a spirocyclic system, can lead to interactions with other molecules or surfaces during lyophilization.

[0005] Not all drugs can be lyophilized. Some drugs may be unstable during lyophilization, or may undergo chemical or physical changes that render them ineffective or unsafe. Additionally, some drugs may have specific formulation requirements or be in forms that make lyophilization challenging. Liposome formulations are a classic example of this.

[0006] Therefore, there is a demand for lyophilized formulations of cryptocephalin. This application is specifically aimed at this need. Summary of the Invention

[0007] The applicant has surprisingly and unexpectedly discovered that cryptocephalin or a pharmaceutically acceptable salt thereof can be lyophilized. This application relates to lyophilized formulations comprising (a) cryptocephalin or a cryptocephalin analogue (e.g., hydroxyurea methyl fumonisin) or a pharmaceutically acceptable salt thereof; and cyclodextrin.

[0008] One aspect includes a lyophilized pharmaceutical compound comprising cryptocephalin, mannitol, water, cyclodextrin, and an organic solvent. This formulation produces a stable lyophilized product. Cryptocephalin is the active pharmaceutical ingredient, mannitol acts as a cryoprotectant and filler, and cyclodextrin is stabilized by forming a complex with cryptocephalin, thereby improving its solubility and stability. The organic solvent helps dissolve the components prior to lyophilization, ensuring uniform distribution of the components. In one example, the composition contains sodium Betadex sulfobutyl ether.

[0009] On the other hand, it includes a lyophilized pharmaceutical compound in which mannitol is present in an amount ranging from about 0.2% to 0.4% by weight, acting as both a filler and a cryoprotectant.

[0010] On the other hand, it includes a lyophilized pharmaceutical compound containing a water content of about 0.1% to 3% by weight. This residual water helps balance product stability and ease of reconstitution.

[0011] On the other hand, it includes a lyophilized pharmaceutical compound having about 50% to 90% by weight of cyclodextrin, which enhances the solubility and stability of cryptocephalosporin.

[0012] On the other hand, it includes a lyophilized pharmaceutical compound containing an organic solvent selected from alcohols, ethers, esters, ketones, and hydrocarbons, which promotes the dissolution of the active pharmaceutical ingredient and other components prior to lyophilization.

[0013] On the other hand, it includes a lyophilized pharmaceutical compound having an organic solvent present in an amount ranging from about 50% to 90% by weight, ensuring complete dissolution of the formulation components.

[0014] On the other hand, this includes lyophilized pharmaceutical compounds using pharmaceutically acceptable grades of mannitol that meet regulatory standards for safety and efficacy.

[0015] On the other hand, it includes a lyophilized pharmaceutical compound that may include additional pharmaceutically acceptable excipients to improve stability, solubility, and overall performance.

[0016] On the other hand, it includes a lyophilized composition that may contain pharmaceutically acceptable excipients such as sodium phosphate, potassium phosphate, citric acid, tartaric acid, gelatin, glycine, mannitol, lactose, sucrose, maltose, glycerol, dextran, dextran, trehalose, hydroxyethyl starch, or combinations thereof, to enhance stability, prevent degradation, and optimize resolvation.

[0017] On the other hand, there is a method for preparing a lyophilized pharmaceutical compound, which involves dissolving the pharmaceutical compound in an organic solvent, followed by adding mannitol, water, and cyclodextrin to the pharmaceutical solution. The mixture is then lyophilized to remove the water content under controlled vacuum conditions to obtain a stable lyophilized product.

[0018] On the other hand, there is a method that involves adding a pharmaceutically acceptable excipient to the drug solution prior to freeze-drying to ensure optimal stability and functionality.

[0019] On the other hand, it includes a lyophilized pharmaceutical compound having hydroxyurea methyl fumonisin, a derivative of cryptocephalin known for its potent anticancer properties, formulated in the lyophilized product to enhance its therapeutic efficacy and stability. Attached Figure Description

[0020] Figure 1 The freeze-drying history curve of (-)-hydroxyurea methyl acyl fulvin (LP-184) is illustrated.

[0021] Figure 2 The results of long-term stability tests on LP-184 are presented.

[0022] Figure 3 Further results from long-term stability testing of the LP-184 are provided.

[0023] Figure 4 The results of the accelerated stability test from LP-184 are shown.

[0024] Figure 5 Additional results from long-term stability tests of the LP-284 are shown.

[0025] Figure 6 Further results from the accelerated stability test of the LP-284 are presented. Detailed Implementation

[0026] This document discloses methods, compositions, and kits for providing solutions and lyophilized formulations of cryptocephalins (including cryptocephalin analogs). Cryptocephalins are generally poorly soluble in water, resulting in low pharmacologically relevant concentrations. Compounds of the cryptocephalin family are unstable during storage and transport at room temperature when formulated in organic media or mixtures of organic and aqueous media. The methods, compositions, and kits provided herein provide pharmaceutically acceptable formulations, including solutions and lyophilized formulations, that address the solubility and stability issues commonly encountered with such compounds. Specifically, the present invention relates to lyophilized formulations comprising (a) hydroxyurea methyl acyl fulvene or a pharmaceutically acceptable salt thereof, mannitol, and cyclodextrin. Methods for preparing and using these lyophilized formulations are also described.

[0027] Cryptocephalins and their analogues face challenges in lyophilization due to their inherent chemical properties. Cryptocephalins exhibit limited solubility in water, making it difficult to formulate them into stable aqueous solutions for lyophilization, as they tend to precipitate or form unstable suspensions. Furthermore, cryptocephalins are sensitive to degradation under conditions such as heat, light, and pH changes. The complex chemical structure of cryptocephalins, including hydroxyurea methyl acyl fulvin with multiple hydroxyl groups and a spirocyclic system, can lead to interactions with other molecules or surfaces during the lyophilization process.

[0028] The term "cryptocephalosin" can refer to several different compounds, generally describing a group of naturally occurring toxins produced by certain fungi in the genus *Omphalotusgenus* and related genera. These toxins are known for their extreme toxicity, and their potential applications in cancer therapy have been investigated. Despite their toxicity, the potential of cryptocephalosporins as anticancer agents has been studied, with research investigating their ability to selectively target and kill cancer cells. Some studies suggest that cryptocephalosporins may have potential as chemotherapeutic agents, particularly in treating certain types of cancer.

[0029] Cryptocephalin is characterized by its unique chemical structure, consisting of fused six-membered and five-membered rings. This structural feature contributes to its biological activity. Analogous compounds are chemical variants or derivatives of the parent cryptocephalin compound that have been modified to potentially enhance its properties or reduce its toxicity. In this context, "cryptocephalin" and its analogues refer to a group of compounds with potential medical or therapeutic properties.

[0030] Cryptocephalin or acyl-richene

[0031] One embodiment of this application includes the use of cryptocephalin or cryptocephalin analogs such as acylfulne. Acylfulne analogs are semi-synthetic derivatives of cryptocephalin, which are cytotoxic and can be extracted from jack-o'-lantern mushroom (Omphalotus olearius). Acylfulne derivatives derived from the sesquiterpene cryptocephalin S via a reverse Prins reaction with an acid are far less reactive with thiols than cryptocephalin S.

[0032] In one example, the acylfulne analogue is (-)-hydroxyureamethylacylfulne (referred to as LP-184 by Lantern Pharma Inc.), which causes light to be negatively deflected, as shown below:

[0033]

[0034] In another example, the acylfulne analogue is (+)-hydroxyurea methyl acylfulne (referred to as LP-284 by Lantern Pharma Inc.), which causes light to be positively deflected, as shown below:

[0035]

[0036] (+)-hydroxyurea methyl acyl fulene and (-)-hydroxyurea methyl acyl fulene are both enantiomers, which are now known.

[0037] Another example includes hydroxymethyl acylfulne, also known as Irofulven.

[0038] Another embodiment includes a lyophilized formulation comprising (a) hydroxyurea methyl acyl fulvene or a pharmaceutically acceptable salt thereof, and Betadex sodium sulfobutyl ether. Methods for preparing and using these lyophilized formulations are also described.

[0039] In other embodiments, various polymers can be used as alternatives to cyclodextrins in lyophilized pharmaceuticals. Examples include polyvinylpyrrolidone (PVP), a water-soluble polymer that can improve the solubility of hydrophobic compounds; hydroxypropyl methylcellulose (HPMC), a cellulose-based polymer that can be used as a cryoprotectant and stabilizer during lyophilization; or dextran, a polysaccharide used to enhance the stability of lyophilized formulations, particularly for proteins and peptides.

[0040] One implementation involves using Betadex sodium sulfobutyl ether, also known as sodium sulfobutyl ether β-cyclodextrin (SBE-β-CD), a chemically modified derivative of β-cyclodextrin. SBE-β-CD is widely used as a complexing agent in pharmaceutical formulations to improve the solubility, stability, and bioavailability of certain drug compounds. Cyclodextrins are cyclic oligosaccharides with a hydrophobic lumen and a hydrophilic outer surface that form inclusion complexes with various drug molecules.

[0041] Another implementation involves formulating hydroxyurea methyl acyl fulvene as a lyophilized powder for injection. The product should be reconstituted as close as possible to the time of patient administration. For the 15 mg product, reconstitution is achieved by aseptically adding 5 mL of sterile water for injection USP (SWFI). For the 100 mg product, use 20 mL of SWFI. Shake the vial thoroughly, and complete dissolution should occur within 5 minutes. After dissolution in SWFI, aseptically remove the required volume of the appropriate dose and immediately transfer it to a 500 mL infusion bag containing either 0.9% sodium chloride injection USP (normal saline) or 2.5% glucose / 0.45% sodium chloride injection USP. Transfer to the infusion bag must be completed within 30 minutes of reconstitution to ensure product stability and efficacy.

[0042] This invention also applies to pharmaceutically acceptable salts of hydroxyurea methyl acyl fulvene. A "pharmaceutically acceptable salt" refers to a derivative of the disclosed compound, wherein the parent compound is modified by forming an acid salt or a base salt. These salts can be, but are not limited to, mineral or organic acid salts of basic residues (such as amines), or base or organic salts of acidic residues (such as carboxylic acids). Pharmaceutically acceptable salts include non-toxic salts or quaternary ammonium salts formed from non-toxic inorganic or organic acids. Preferred examples include salts derived from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, citric acid, and other pharmaceutically acceptable acids.

[0043] Specific embodiments cover lyophilized formulations of hydroxyurea methyl acyl fulvene or its pharmaceutically acceptable salts in combination with cyclodextrin. Cyclodextrin is a cyclic oligosaccharide containing five or more α-D-glucopyranoside units, which can enhance the solubility and stability of many drugs, including hydroxyurea methyl acyl fulvene. The cyclodextrins of the present invention include naturally occurring and derived forms, such as hydroxypropyl-β-cyclodextrin or sulfobutyl ether β-cyclodextrin, which are preferred due to their solubilizing properties.

[0044] Cyclodextrin derivatives such as methyl-β-cyclodextrin, dimethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and sulfobutyl ether β-cyclodextrin are particularly effective in stabilizing hydroxyurea methyl acyl fulvin during and after lyophilization. These cyclodextrins can be present in amounts up to 99% w / w of the lyophilized formulation, with a preferred concentration range of 20% w / w to 90% w / w. The most preferred embodiment contains 40% w / w to 70% w / w of cyclodextrin.

[0045] Besides cyclodextrins, lyophilized formulations may also contain fillers to enhance the structure of the resulting cake. Fillers such as mannitol are commonly used in the lyophilization process of pharmaceuticals to strengthen the lyophilized product, making it more stable and easier to handle. Mannitol can be present at concentrations ranging from 20% w / w to 60% w / w in the lyophilized formulation.

[0046] In some implementations, pharmaceutically acceptable organic solvents (such as ethanol, tert-butanol, acetone, ethyl acetate, or heptane) are used to prepare the pre-lyophilized solution. These solvents facilitate the dissolution of hydroxyurea methyl acyl fulvene, making the lyophilization process more efficient. After lyophilization, the residual solvent is reduced to less than 3% w / w, ensuring the safety and efficacy of the final product.

[0047] The lyophilized formulation can be rapidly reconstituted, forming a clear, colorless solution free of particulate matter within 180 seconds. In some preferred embodiments, reconstitution occurs in less than 60 seconds, providing a significant improvement over existing formulations.

[0048] Lyophilized cakes of hydroxyurea methyl acyl-richene can contain varying concentrations of the active ingredient, depending on the intended therapeutic use and formulation requirements. Typically, the concentration of hydroxyurea methyl acyl-richene in the lyophilized cake ranges from about 2.5 mg / mL to 30 mg / mL or from 2.5 mg / mL to 25 mg / mL. In one example, the cake has a concentration of 15 mg / mL. This range allows for flexibility in administration while maintaining the stability and efficacy of the drug. The concentration can be controlled to ensure the lyophilized cake is stable throughout its shelf life and can be easily reconstituted for administration. In specific formulations, the concentration can be adjusted based on the desired therapeutic effect, the solubility of the active ingredient, and compatibility with excipients such as mannitol and cyclodextrin. These excipients play a role in maintaining the structural integrity of the lyophilized cake while enhancing the solubility and stability of hydroxyurea methyl acyl-richene.

[0049] These lyophilized formulations can also be used in combination with other antitumor agents. Combination therapy is widely used to treat neoplastic diseases, and hydroxyurea methyl fumonisin can be used in combination with agents such as glucocorticoids, interferon, or other anticancer drugs to enhance therapeutic efficacy.

[0050] Therefore, the present invention provides significant progress in the formulation of hydroxyurea methyl acyl fulvin, improving its solubility, stability and ease of use in clinical applications.

[0051] One embodiment includes a method for manufacturing the lyophilized solid composition of hydroxyurea methyl acyl fulene, the method involving dissolving the hydroxyurea methyl acyl fulene in an organic lyophilizing solvent and subjecting the solution to a first lyophilization. The resulting cake containing up to 8% w / w of the organic lyophilizing solvent is then reconstituted with an aqueous solution and subjected to a second lyophilization. The method may include the following steps:

[0052] a) Dissolve hydroxyurea methyl acyl fulvin in an organic lyophilizing solvent.

[0053] b) Freeze-drying,

[0054] c) Dissolve the lyophilized hydroxyurea methyl acyl fulvene from step (b) in an aqueous solution, and

[0055] d) Lyophilize the aqueous solution of hydroxyurea methyl acyl fulvin from step (c).

[0056] Lyophilization offers several advantages for drug preservation and storage. By removing moisture, it minimizes the potential for microbial growth and chemical degradation, extending the shelf life of the drug. Additionally, lyophilized products can be easily reconstituted with a suitable solvent, such as sterile water, before being administered to patients.

[0057] The optimal freeze-dried cake should have a uniform appearance, adhere to the vial wall without detaching, and have no cavities at the bottom. The color of the cake should be consistent, although slight variations are possible due to crystal formation during the freezing process, whether dendritic or small and amorphous. In one example, a combination of mannitol and cyclodextrin provided such a cake.

[0058] Lyophilized formulations of hydroxymethyl acyl-rich olefins typically contain a variety of excipients, such as buffers, stabilizers, fillers, and tension modifiers. Well-chosen excipient formulations ensure that lyophilized products have a long shelf life and stable biological activity. Excipients play a crucial role in drug product development to achieve the desired product profile, including stability and efficacy.

[0059] One advantage of these specific implementation schemes is that they provide simple and stable freeze-dried formulations while maintaining acceptable chemical, physical, and microbiological stability.

[0060] In formulations containing low concentrations of cryptocephalin, fillers help to produce larger, more robust cake structures. Crystalline fillers are commonly used to produce well-formed cakes with good mechanical properties. In lyophilized formulations, fillers such as mannitol and glycine are used to provide structure to the lyophilized cakes and prevent collapse.

[0061] Other components include cryoprotectants or lyophilization protectants, which are used to protect biological materials such as cells, tissues, proteins, and vaccines from freezing (cryoprotectants) or drying (lyophilization protectants). These substances are essential for maintaining the activity and functionality of these materials during long-term storage or transportation. Excipients such as sugars (e.g., sucrose, trehalose), sugar alcohols (e.g., mannitol, sorbitol), and surfactants (e.g., polysorbate 20, 80) are typically used to improve stability.

[0062] Tension regulator

[0063] Strain modifiers are typically selected based on the stability requirements of the bulk solution or the route of administration. Excipients such as mannitol, sucrose, glycine, glycerol, and sodium chloride are commonly used as strain modifiers. These excipients can be included in the diluent rather than in the formulation itself and play an important role in enhancing the rapid solubility and absorption of lyophilized products.

[0064] The following embodiments are provided for illustrative purposes only and should not be construed as limiting the scope of the invention.

[0065] Example

[0066] Example 1 - Lyophilized (-)-hydroxyurea methyl acyl fulvene (LP-184)

[0067] The lyophilized composition consists of several key components. LP-184 is the main active ingredient, present at a concentration of 2.5 mg / mL. Mannitol, at 83.3 mg / mL, acts as a filler to help maintain product integrity during lyophilization, a critical part of the manufacturing process. Betadex sodium sulfobutyl ether, present at a concentration of 166.7 mg / mL, acts as a complexing agent to improve solubility and stability. Water is used to bring the formulation to 99%, and the water is subsequently removed during the lyophilization process to ensure a stable dried product.

[0068] The preparation of this composition involves several steps. First, Betadex sodium sulfobutyl ether (BSES) is added to a vessel and heated to a temperature between 60°C and 65°C. Then, the solution is cooled to approximately 25°C in preparation for the addition of other components. LP-184 is added, followed by mannitol. Water is then introduced to achieve a final composition of 99%.

[0069] Once the solution is prepared, it undergoes a freeze-drying process. The temperature is lowered to below 15°C under vacuum to remove water content. The material is frozen for up to 240 minutes to stabilize it before primary drying. During the primary drying stage, the pressure is maintained between 70 and 200 mTorr, with drying cycles lasting over 1500 minutes. If necessary, a secondary drying step is performed, during which the product is dried at a higher temperature (typically 50 mTorr) for over 360 minutes. This freeze-drying process is crucial for ensuring the stability of the freeze-dried product, which can be reconstituted when needed.

[0070] Figure 1 The lyophilization history curve for (-)-hydroxyurea methyl acyl fulvene is shown. Lyophilization history curves, or lyophilization process curves, are graphical representations used to monitor and control key stages of the lyophilization process in pharmaceutical and biotechnology manufacturing. These curves illustrate the relationships between time, temperature, pressure, and other parameters during the three main stages of lyophilization: freezing, primary drying, and secondary drying. They help visualize how the product, shelf, and chamber temperatures evolve over time, thus ensuring optimal conditions and a stable final product.

[0071] Example 2 - Lyophilized (+)-hydroxyurea methyl acyl fulvene (LP-284)

[0072] The lyophilized composition is designed to ensure stability and efficacy. LP-284, one of the active components, is present at a concentration of 2.5 mg / mL. Mannitol, contained at 83.3 mg / mL, acts as a filler to protect the formulation during lyophilization, serving as both a stabilizer and a cryoprotectant. Sulfobutyl ether β-cyclodextrin, present at 166.7 mg / mL, acts as a complexing agent to enhance the solubility and stability of LP-284. Water is added as a solvent to bring the composition to 99%, and the water is subsequently removed during lyophilization to ensure product stability.

[0073] The preparation method for this composition involves several key steps. The method begins with dissolving sulfobutyl ether β-cyclodextrin in a vessel at a temperature between 60°C and 65°C. Once dissolved, the solution is cooled to approximately 25°C in preparation for the addition of other components. LP-284 is then added, followed by mannitol, which causes the formulation to swell and protects it during freeze-drying. Water is then added to achieve the desired final composition.

[0074] Once the solution is prepared, it is freeze-dried to remove water and stabilize the product. The solution is cooled to below 15°C, and vacuum conditions are applied to promote water sublimation. The material is initially frozen for up to 240 minutes, followed by primary drying at pressures ranging from 70 mTorr to 200 mTorr, which can last for more than 1500 minutes. A secondary drying step can be used to further reduce the moisture content. This involves drying at higher temperatures (approximately 50 mTorr) for more than 360 minutes, depending on the product's stability requirements.

[0075] Example 3: Stability of freeze-dried LP-184

[0076] As demonstrated by the 12-month stability test conducted at both 25°C / 60%RH (relative humidity) and 2°C–8°C, the lyophilized product of Example 1 remained stable over a one-year period. The concentration of LP-184 in the product was 15.0 mg / mL, and the formulation showed no significant degradation or loss of potency during this period. To ensure product reliability, clinical batches must undergo both accelerated stability testing and long-term stability testing as part of regulatory and quality assurance requirements.

[0077] For these studies, samples were stored upright in qualified stability storage rooms and cabinets. Temperature and humidity were continuously monitored to ensure compliance with specified conditions. Throughout the testing, the products exhibited stability when stored at 25°C (77℉), with no observable changes in appearance, efficacy, or integrity.

[0078] Figure 2 and Figure 3 The results of long-term stability studies are illustrated, demonstrating the product's consistency under normal storage conditions. Additionally, Figure 3 The results of accelerated stability studies are presented, in which the product is exposed to more extreme conditions to simulate long-term effects over shorter time periods. These studies confirm the robustness of the formulation, validate its suitability for clinical use, and ensure its compliance with regulatory stability requirements.

[0079] Example 4: Stability of freeze-dried LP-284

[0080] As demonstrated by 12-month stability tests conducted at both 25°C / 60%RH and 2°C–8°C, the freeze-dried product (LP-284) of Example 2 remained stable over a one-year period. This test confirms the product's ability to maintain its physical and chemical properties under both standard and refrigerated storage conditions, highlighting its robustness for long-term use.

[0081] To comply with regulations, clinical batches of products are required to undergo both accelerated stability testing and long-term stability testing to assess their behavior under various environmental conditions. These tests are essential for understanding how the product will perform throughout its shelf life and ensuring it meets all safety and efficacy criteria. During the testing period, samples are stored upright in qualified stability storage rooms and cabinets designed to maintain consistent conditions. Temperature and humidity are continuously monitored and recorded using a validated system to ensure precise control of the storage environment.

[0082] Stability testing confirmed that the product maintained its integrity and stability when stored at 25°C (77℉), without significant reduction in potency, appearance, or other key quality attributes. Ensuring product stability throughout its intended shelf life is crucial for delivering consistent therapeutic benefits.

[0083] Figure 4 The results of the long-term stability study are summarized, confirming that the key characteristics of the product are retained over time. Additionally, Figure 5 The results of an accelerated stability study are presented, in which the product is exposed to stress conditions to simulate the effects of long-term storage over a shorter timeframe. The findings from the accelerated study further support the product's stability, demonstrating its resilience to environmental challenges during storage and transportation. This provides confidence in the product's long-term reliability.

[0084] These comprehensive stability tests are crucial in ensuring the quality, safety, and efficacy of products, and in verifying their suitability for both clinical and commercial use.

[0085] As those skilled in the art will understand, many modifications and variations of the invention are possible in light of the foregoing teachings. Therefore, it should be understood that the invention may be practiced in ways other than those specifically described herein, within the scope of the appended claims, and the scope of the invention is intended to cover all such variations.

Claims

1. A freeze-dried composition comprising cryptocephalin, mannitol, water, cyclodextrin, and an organic solvent.

2. The freeze-dried composition according to claim 1, wherein the cryptocephalin is hydroxyurea methyl acyl fulvene.

3. The freeze-dried composition according to claim 1, wherein the mannitol is present in an amount ranging from about 0.2% by weight to 0.4% by weight.

4. The freeze-dried composition according to claim 1, wherein the water is present in an amount ranging from about 0.1% by weight to 3% by weight.

5. The freeze-dried composition according to claim 1, wherein the cyclodextrin is present in an amount ranging from about 50% to 90% by weight.

6. The freeze-dried composition according to claim 1, wherein the organic solvent is selected from the group consisting of alcohols, ethers, esters, ketones and hydrocarbons.

7. The freeze-dried composition according to claim 1, wherein the organic solvent is present in an amount ranging from about 50% to 90% by weight.

8. The lyophilized composition according to claim 1, wherein the lyophilized composition further comprises a pharmaceutically acceptable grade of mannitol.

9. The lyophilized composition according to claim 1, wherein the lyophilized composition further comprises a pharmaceutically acceptable excipient.

10. The freeze-dried composition according to claim 9, wherein the pharmaceutically acceptable excipient is selected from the group consisting of sodium phosphate, potassium phosphate, citric acid, tartaric acid, gelatin, glycine, mannitol, lactose, sucrose, maltose, glycerol, dextran, dextran, trehalose, hydroxyethyl starch, or mixtures thereof.

11. A method for preparing the lyophilized composition according to claim 1, the method comprising: a. Dissolve the drug compound in the organic solvent, b. Add mannitol, water and cyclodextrin to the drug solution, and c. subject the mixture to freeze-drying.

12. The method according to claim 11, wherein the cryptocephalin is hydroxyurea methyl acyl fulvin.

13. The method of claim 11, further comprising adding a pharmaceutically acceptable excipient to the drug solution prior to freeze-drying.

14. A pharmaceutical composition comprising the lyophilized composition according to claims 1 to 9 and a pharmaceutically acceptable carrier or diluent.

15. The freeze-dried composition according to claims 1 to 9, wherein the cryptocephalin is hydroxyurea methyl acyl fulvene.

16. A method for manufacturing a lyophilized composition, the method comprising: a. Add cyclodextrin to the solution and heat to a temperature between 60°C and 65°C. b. Cool the solution. c. Add cryptocephalin to the cooled solution, and d. Add mannitol to the solution.

17. The method of claim 16, wherein the cryptocephalin is hydroxyurea methyl fumonisin.

18. The method of claim 16, further comprising introducing water to achieve a final composition of up to 99%.

19. The method of claim 16, wherein the concentration of LP-184 in the composition is between 2.5 mg / mL and 25 mg / mL.

20. The method of claim 16, further comprising the step of freeze-drying the prepared solution.

21. The method of claim 20, wherein the freeze-drying process further comprises a secondary drying step, wherein the pressure is maintained at approximately 50 mTorr for more than 360 minutes to further remove residual moisture.

22. The method of claim 16, wherein the freeze-dried composition produced by the method is stable over a period of 12 months when stored at a temperature of 25°C.

23. A method for treating a subject requiring hydroxyurea methyl acyl-richene, the method comprising administering to the subject a therapeutically effective amount of a lyophilized formulation of hydroxyurea methyl acyl-richene, wherein the lyophilized formulation is reconstituted in a pharmaceutically acceptable aqueous solution prior to administration.

24. The method of claim 23, wherein the administration is performed by intravenous infusion over a period of 30 to 60 minutes, thereby treating the subject.

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