Polycyclic compound injection and preparation method therefor

Abstract

A polycyclic compound injection and a preparation method therefor. The present invention pertains to the field of pharmaceutical formulations for cancer. The polycyclic compound injection comprises a polycyclic compound represented by formula I, low-viscosity polyethylene glycol, Tween 80, and polyoxyethylene (35) castor oil, wherein the concentration of the polycyclic compound represented by formula I is 0.1-15 mg / mL. In addition, the present invention further relates to a ready-to-use polycyclic compound injection comprising water, a polycyclic compound injection, and an osmotic pressure regulator, wherein the ready-to-use polycyclic compound injection has a pH of 3.2-6.5 and is isotonic. The polycyclic compound injection improves the solubility of the polycyclic compound represented by formula I in aqueous solutions while demonstrating good stability, good auxiliary material tolerability or safety in the body, low costs, ease of production, and good tolerance to storage. The polycyclic compound injection is suitable for large-scale clinical experiments and research.

Description

Polycyclic compound injections and their preparation methods Technical Field

[0001] This invention belongs to the field of cancer drug formulations, specifically relating to an injectable formulation of a polycyclic compound and its preparation method. Background Technology

[0002] DHX33 belongs to the family of RNA helicase proteins containing the DEAD / H box, where DEAD / H stands for the amino acid abbreviation Asp-Glu-Ala-Asp / His. This sequence, along with several other conserved amino acid sequences, appears in the protein sequences of RNA helicase family members and is highly involved in nucleic acid substrate binding and ATP hydrolysis. Although these family members share these common sequences, each RNA helicase has its own specificity and unique biological function. The human DHX33 protein has a molecular weight of 72 kDa and the function of unwinding nucleic acids. It uses the bioenergy released from ATP hydrolysis to drive changes in the conformation of RNA and protein complexes, thereby participating in a series of RNA metabolic activities, specifically from RNA transcription, splicing, editing, translation to degradation. The function of DHX33 is not limited to the modification of RNA molecules. Studies have shown that in addition to unwinding RNA double strands, DHX33 also participates in DNA metabolism. Specifically, DHX33 can unwind the DNA double-strand structure and plays an important role in gene expression. In in vitro enzyme reaction systems, DHX33 has also been found to untangle the hybrid double-stranded structure of DNA / RNA.

[0003] Studies have shown that DHX33, by binding to promoters of various cancer-related genes, affects DNA methylation status, thereby regulating the expression of multiple oncogenes and tumor development-related signaling pathways at the genomic level. It plays a crucial role in various cellular activities, including cell growth, proliferation, migration, apoptosis, and glucose metabolism. Furthermore, DHX33 has been found to sense the invasion of foreign double-stranded RNA molecules and play an important role in cellular innate immunity. As a vital cell growth regulator, DHX33 is highly expressed in various cancers. The development and progression of many cancers depend on the high expression of DHX33 protein. Genetic knockout of DHX33 can significantly inhibit the development and progression of RAS oncogene-driven lung cancer; in vivo and in vitro experiments have confirmed that inhibition of DHX33 protein significantly suppresses the development and progression of various cancers, such as breast cancer, colon cancer, glioma, and lymphoma.

[0004] Studies have shown that the protein function of DHX33 depends on its helicase activity. DHX33 helicase-deficient mutants do not possess the function of the DHX33 protein and cannot replace the function of the wild-type DHX33 gene.

[0005] Chinese patent ZL 202110724793.1 and Chinese patent applications 202211413921.1, 202211486053.X, and 202211486081.1 all disclose inhibitors of DHX33, such as the polycyclic compound KY386. However, KY386 has poor water solubility, with an equilibrium solubility of less than 0.01 mg / mL in water after 24 hours, classifying it as a poorly soluble compound. Therefore, improving the water solubility of this inhibitor while ensuring it possesses properties that meet the requirements of clinical trials, including stability, ease of production, low formulation cost, high tolerability, and ease of transportation and storage, is a pressing issue that needs to be addressed.

[0006] Summary of the Invention

[0007] In view of this, the purpose of the present invention is to provide a polycyclic compound injection and a preparation method thereof, so as to solve the problem of poor water solubility of polycyclic compounds in the prior art, and at the same time provide a stable and readily tolerated injectable.

[0008] On the one hand, the present invention provides a polycyclic compound injection containing a polycyclic compound of formula I, low-viscosity polyethylene glycol, Tween 80 (also known as polysorbate 80), and polyoxyethylene (35) castor oil, wherein the concentration of the polycyclic compound of formula I is 0.1-15 mg / mL.

[0009] In an embodiment of the invention, the injection contains about 14.28 mg / mL of the polycyclic compound of formula I.

[0010] In an embodiment of the present invention, the volume ratio or mass ratio of low-viscosity polyethylene glycol, Tween 80 and polyoxyethylene (35) castor oil is about 2:2:1.

[0011] In an embodiment of the present invention, the low-viscosity polyethylene glycol is PEG300, PEG400, or a mixture thereof.

[0012] In an embodiment of the present invention, the polyoxyethylene (35) castor oil can be polyoxyethylene (35) castor oil EL or polyoxyethylene (35) castor oil ELP.

[0013] In a preferred embodiment of the present invention, the injection contains a polycyclic compound of formula I, PEG300, Tween 80 and polyoxyethylene (35) castor oil, wherein the concentration of the polycyclic compound of formula I is 0.1-15 mg / mL, preferably 14.28 mg / mL, and the volume ratio or mass ratio of PEG300, Tween 80 and polyoxyethylene (35) castor oil is about 2:2:1.

[0014] In an embodiment of the present invention, the injection is not mixed with water or its water content is controlled within 1% by mass.

[0015] In a second aspect, the present invention provides a method for preparing a polycyclic compound injection, the method comprising the following steps:

[0016] S1. After heating the low-viscosity polyethylene glycol to 60±5℃, keep the temperature constant and keep it under nitrogen (0.03Mpa) for at least 20 minutes;

[0017] S2. Under stirring conditions, add the polycyclic compound shown in Formula I and stir until fully dissolved;

[0018] S3. Add Tween 80, maintain nitrogen pressure (0.03 MPa) and stir thoroughly for 10 minutes; and

[0019] S4. Add polyoxyethylene (35) castor oil, continue stirring and nitrogen purging for 30 minutes until the mixture is homogeneous, thus obtaining the polycyclic compound injection.

[0020] In an embodiment of the second aspect of the present invention, the concentration of the polycyclic compound shown in Formula I is about 0.1-15 mg / mL.

[0021] In an embodiment of the second aspect of the present invention, the volume ratio or mass ratio of low-viscosity polyethylene glycol, Tween 80 and polyoxyethylene (35) castor oil is about 2:2:1.

[0022] In an embodiment of the second aspect of the present invention, the low-viscosity polyethylene glycol is PEG300, PEG400, or a mixture thereof.

[0023] In an embodiment of the second aspect of the present invention, the polyoxyethylene (35) castor oil can be polyoxyethylene (35) castor oil EL or polyoxyethylene (35) castor oil ELP.

[0024] In a third aspect, the present invention provides a ready-to-use polycyclic compound injection containing water, a polycyclic compound injection, and an osmotic pressure regulator.

[0025] In an embodiment of the third aspect of the present invention, the concentration of the polycyclic compound shown in Formula I is about 0.1-0.5 mg / mL, the pH of the ready-to-use polycyclic compound injection is 3.2-6.5, and it is isotonic.

[0026] In a fourth aspect, the present invention provides an analytical separation method for the content and impurities of the polycyclic compound of Formula I in the above-mentioned polycyclic compound injectable formulation, wherein the chromatographic conditions are as follows: octadecyl bonded silica gel is used as the packing material (Phenomenex Gemini C18, 4.6 × 250 mm, 5 μm or equivalent column); 0.1% formic acid is used as mobile phase A, methanol is used as mobile phase B, and gradient elution is performed according to the table below; the column temperature is 30 °C; the detection wavelength is 246 nm; the flow rate is 1.0 mL per minute; the injection volume is 10 μL; and the gradient elution time is set to 37 minutes.

[0027] As can be seen from the above embodiments, the present invention provides a polycyclic compound injection that uses low-viscosity polyethylene glycol, Tween 80 and polyoxyethylene (35) castor oil as solvents, which improves the solubility of the polycyclic compound shown in Formula I, and at the same time improves the stability and storage resistance of the injection, making it suitable for clinical trial research. Attached Figure Description

[0028] Figure 1 shows the production process for preparing polycyclic compound injections in a GMP workshop.

[0029] Figure 2 shows the chromatographic standard curve of the polycyclic compound represented by Formula I, where the vertical axis is the peak area and the horizontal axis is the concentration (μg / mL). Detailed Implementation

[0030] On the one hand, the present invention provides a polycyclic compound injection containing 0.1-15 mg / mL of the polycyclic compound of formula I, low-viscosity polyethylene glycol, Tween 80, and polyoxyethylene (35) castor oil.

[0031] In polycyclic aromatic hydrocarbon injections, the content of the polycyclic aromatic hydrocarbon represented by Formula I can be approximately 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 5.0 mg / mL, 6.0 mg / mL, 7.0 mg / mL, 8.0 mg / mL, 9.0 mg / mL, 10.0 mg / mL, 11.0 mg / mL, 12.0 mg / mL, 13.0 mg / mL, 14.0 mg / mL, 14.5 mg / mL, and any value between any two of the above values, such as approximately 0.25 mg / mL or 0.35 mg / mL. In a preferred embodiment, the concentration of the polycyclic compound shown in Formula I is about 14.28 mg / mL.

[0032] In an embodiment of the present invention, in the polycyclic compound injection of the present invention, the volume or mass ratio of low-viscosity polyethylene glycol in the mixed solvent is not less than 40%, the volume or mass ratio of polyoxyethylene (35) castor oil in the mixed solvent is not less than 20%, and the volume or mass ratio of Tween 80 in the mixed solvent is not more than 40%. Preferably, the mixed solvent of the polycyclic compound injection contains, or is composed of, approximately 40% by volume or mass of low-viscosity polyethylene glycol, approximately 40% by volume of Tween 80, and approximately 20% by volume of polyoxyethylene (35) castor oil.

[0033] In the polycyclic aromatic hydrocarbon injection, the volume or mass ratio of low-viscosity polyethylene glycol, Tween 80, and polyoxyethylene (35) castor oil is approximately 2:2:1. In a preferred embodiment, the low-viscosity polyethylene glycol is PEG300, PEG400, or a mixture thereof. In a specific embodiment, the polyoxyethylene (35) castor oil can be polyoxyethylene (35) castor oil EL or polyoxyethylene (35) castor oil ELP.

[0034] In this specification, the term "about" generally refers to + / -5% of the value, more typically + / -4%, more typically + / -3%, more typically + / -2%, even more typically + / -1%, even more typically + / -0.5%.

[0035] In a further preferred embodiment, the polycyclic compound injection contains the polycyclic compound of Formula I, PEG300, Tween 80, and polyoxyethylene (35) castor oil, wherein the concentration of the polycyclic compound of Formula I is about 14.28 mg / mL, and the volume ratio or mass ratio of PEG300, Tween 80, and polyoxyethylene (35) castor oil is about 2:2:1.

[0036] Obviously, to further enhance the stability of polycyclic aromatic hydrocarbon injections, other harmless substances that do not react with the polycyclic aromatic hydrocarbons shown in Formula I can be added. Generally, these substances can be:

[0037] Protective gas is used to significantly reduce the content of reactive gases such as O2 and CO2 in the polycyclic compound injection by evacuating the injection. Then, an inert gas that does not react with the polycyclic compound, polyethylene glycol, or polyoxyethylene (35) castor oil shown in Formula I, such as N2 or other inert gases, is introduced. These protective gases will dissolve in the injection; or

[0038] Antioxidants, such as vitamin E, can be added to polycyclic aromatic hydrocarbon injections to further enhance their stability.

[0039] In embodiments of the present invention, in order to improve patient compliance, the polycyclic compound injection of the present invention may also contain anesthetic or analgesic drugs in doses permitted by pharmacopoeia or formulary.

[0040] In embodiments of the present invention, the polycyclic compound injection may also contain another therapeutic agent to form a compound preparation, thereby synergistically enhancing the therapeutic effect. In particular, these drugs are recommended to be those that regulate the expression level of the DHX33 enzyme or its corresponding gene.

[0041] In some cases, to cope with special environments, the polycyclic compound injections of the present invention may also contain antibacterial agents, antifungal agents, etc.

[0042] Obviously, the amount of protective gases, antioxidants, drugs with anesthetic or analgesic effects, antibacterial agents, antifungal agents, etc. added should not affect the solubility or other properties of the polycyclic compound shown in Formula I, or enhance its stability; while the content of another therapeutic agent, substances that regulate water-salt, electrolyte and acid-base balance, osmotic pressure, and pH-regulating substances need to be determined according to the purpose, prescription dosage, pharmacopoeia, formulary, etc., based on the actual situation.

[0043] Obviously, the polyethylene glycol and polyoxyethylene (35) castor oil specified in this invention should each be liquid at room temperature and differential pressure, or the mixed solvent should be liquid. Both polyoxyethylene (35) castor oil and polyethylene glycol in the polycyclic aromatic hydrocarbon injection have high viscosity, resulting in a high viscosity for the injection. PEG300 has a slightly lower viscosity than PEG400, therefore PEG300 is preferred, as it reduces the overall viscosity of the injection. From another perspective, these substances also hinder the movement of active ingredient molecules, thus enhancing the stability of the polycyclic aromatic hydrocarbon injection.

[0044] Since the body may have an allergic reaction to excessive amounts of polyoxyethylene (35) castor oil, and the purity of polyoxyethylene (35) castor oil ELP is higher than that of polyoxyethylene (35) castor oil EL, polyoxyethylene (35) castor oil ELP is preferred.

[0045] In embodiments of the present invention, the polycyclic compound injection is anhydrous, or its water content is controlled to be less than 1% by mass. Obviously, "no water added" means that no additional water is added during the preparation process, and other reagents and solvents used are anhydrous reagents, such as anhydrous low-viscosity polyethylene glycol. In other words, the water content needs to be controlled during the preparation of this injection. Theoretically, the lower the water content of the injection, the better its stability. However, due to technical difficulties and considerations for large-scale industrial production, it is preferable that the water content (determined by Karl Fischer method) is within 1% by mass.

[0046] As a specific implementation, the polycyclic compound injection contains 1.4 mL of PEG300 and 1.36 mL of injection-grade Tween 80, 0.7 mL of polyoxyethylene (35) castor oil and 50 mg of the polycyclic compound of formula I, or is composed thereof.

[0047] In an embodiment of the present invention, a polycyclic compound injection can be filled into a 10 mL volume borosilicate glass tube (brown) injection bottle, then filled with an inert protective gas, capped with a rubber stopper, and sealed with an aluminum-plastic composite cap for antibiotic bottles.

[0048] Clearly, the 3.9 mL dosage form is slightly higher than the standard 3.5 mL. This is because, in clinical trials, some syringe residue may remain after direct extraction. Therefore, the dosage is increased to ensure the extraction volume meets the actual specifications of the formulation. Reasonable volume variations are well-known to those skilled in the art.

[0049] In a second aspect, the present invention provides a method for preparing a polycyclic compound injection, the method comprising the following steps:

[0050] S1. Add low-viscosity polyethylene glycol to the preparation container, heat the above excipients to 60±5℃, keep the temperature constant, and keep nitrogen blowing (0.03Mpa) for at least 20 minutes;

[0051] S2. Under stirring conditions, add the polycyclic compound shown in Formula I to the above excipients and stir until fully dissolved;

[0052] S3. Stop heating, then add Tween 80, maintain nitrogen purging (0.03 MPa) and stir thoroughly for 10 minutes; and

[0053] S4. Then add polyoxyethylene (35) castor oil, continue stirring and nitrogen purging for 30 minutes until the mixture is homogeneous, which is the polycyclic compound injection.

[0054] This procedure is based on the viscous, oily properties of the polycyclic compound shown in Formula I. Preferably, the first excipient added is PEG300, which is easy to weigh, transfer, heat, and dissolve the polycyclic compound shown in Formula I.

[0055] The polycyclic compound injection of the present invention is a concentrated form, which needs to be diluted with a solution prepared by a medically acceptable osmotic pressure regulator (as solute) and water (as solvent) before administration.

[0056] Therefore, in a third aspect, the present invention provides a ready-to-use polycyclic compound injection containing water, the aforementioned polycyclic compound injection, and an osmotic pressure regulator.

[0057] In an embodiment of the third aspect of the present invention, the concentration of the polycyclic compound shown in Formula I is about 0.1-0.5 mg / mL, the pH of the ready-to-use polycyclic compound injection is 3.2-6.5, and it is isotonic.

[0058] Osmotic regulators are well known to those skilled in the art and include, but are not limited to, sodium chloride, glucose, sucrose, fructose, xylitol, glycerol, sorbitol, mannitol, potassium chloride, mannose, calcium chloride, magnesium chloride, and other inorganic salts in anhydrous or aqueous forms. Glucose is a preferred osmotic regulator.

[0059] As is well known to those skilled in the art, isotonic solutions, generally defined for humans, are those with a concentration of 308 mmol / L, such as 0.9% saline and 5% glucose solutions. However, the concentration can be broadly extended to values ​​between 280-350 mmol / L. Of course, if the intended treatment is for other animals, such as other primates, the concentration of the isotonic solution can be adjusted accordingly.

[0060] In a specific implementation of the third aspect, the concentration of the polycyclic compound shown in Formula I can be, for example, about 0.1 mg / mL, 0.15 mg / mL, 0.2 mg / mL, 0.25 mg / mL, 0.3 mg / mL, 0.35 mg / mL, 0.4 mg / mL, 0.45 mg / mL, 0.48 mg / mL, 0.49 mg / mL, 0.5 mg / mL, and any value between any two of the above values, such as 0.13 mg / mL, 0.22 mg / mL, etc.

[0061] As is known to those skilled in the art, substances that adjust pH include alkali metal and alkaline earth metal salts of weak acids such as carbonic acid, phosphoric acid, citric acid, and acetic acid, as well as inorganic bases such as hydroxides of alkali metals and alkaline earth metals such as Na and K. Examples include sodium citrate, potassium citrate, sodium acetate, potassium acetate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, potassium phosphate, sodium phosphate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate, and potassium carbonate.

[0062] In the specific implementation plan of the third aspect, the pH of the ready-to-use polycyclic aromatic hydrocarbon injection is 3.2-6.5, for example, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, and any value between two of the above. Ready-to-use polycyclic aromatic hydrocarbon injections within this pH range exhibit good stability, ensuring at least 3 hours of room temperature stability, thus preventing situations where patients cannot take the medication on time after on-site preparation.

[0063] Ready-to-use polycyclic aromatic hydrocarbon injections can be prepared in the following manner: For example, the method for preparing a polycyclic aromatic hydrocarbon injection as a ready-to-use intravenous injection is as follows: dilute 3.5 mL (50 mg of the polycyclic aromatic hydrocarbon shown in Formula I) of the polycyclic aromatic hydrocarbon injection with 100 mL of 5% glucose injection and adjust the pH of the mixed solution to the range of 3.2-6.5; mix and dissolve to obtain the final product.

[0064] In a fourth aspect, this invention provides an analytical separation method for the content of polycyclic compounds and impurities in injectable polycyclic compound formulations. The chromatographic conditions are as follows: octadecyl-bonded silica gel is used as the packing material (Phenomenex Gemini C18, 4.6 × 250 mm, 5 μm or equivalent column); 0.1% formic acid is used as mobile phase A, and methanol as mobile phase B, with gradient elution performed according to the table below; the column temperature is 30°C; the detection wavelength is 246 nm; the flow rate is 1.0 mL per minute; the injection volume is 10 μL; and the gradient elution time is set to 37 minutes. These conditions can effectively separate the target compound (i.e., the polycyclic compound shown in Figure I) from impurities, and determine its content in accordance with the requirements of the Chinese Pharmacopoeia.

[0065] Chromatographic analysis conditions

[0066] Example

[0067] The preferred embodiments of the present invention will now be described in detail with reference to specific examples. It should be understood that the following embodiments are provided for illustrative purposes only and are not intended to limit the scope of protection of the present invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from its spirit and intent, and all such modifications and substitutions fall within the scope of protection claimed in the present invention.

[0068] In the following examples, the polycyclic compound of Formula I (hereinafter referred to as KY386, API, or active pharmaceutical ingredient) was used as the active ingredient, and PEG300, Tween 80, polyoxyethylene (35) castor oil ELP, etc., were used as raw materials and excipients. Unless otherwise specified, the experimental methods used in the following examples are conventional methods, such as referring to the 2020 edition of the Chinese Pharmacopoeia. Unless otherwise specified, the materials and reagents used in the following examples can be obtained commercially.

[0069] 1. Solubility Study of KY386

[0070] 1.1 Buffer / Solution Preparation

[0071] As long as the target concentration remains unchanged, different volumes of stock solutions and buffer solutions than specified can be used.

[0072] Table 1: Preparation methods of aqueous solutions with different pH values

[0073] 1.2 Chromatographic determination of the aqueous solubility of KY386

[0074] 1) Preparation of test solution

[0075] Accurately weigh an appropriate amount of KY386 raw material, dissolve it completely in methanol and dilute it to a final concentration of 10 mg / mL. Take 10 μL of the solution, record the peak area, and calculate the content of KY386 in the sample.

[0076] 2) Examining the linear range of KY386

[0077] Accurately weigh an appropriate amount of KY386 standard (purity 99.9%, content 99.8%), dissolve it in methanol to prepare a 5000 μg / mL solution. Then, gradually dilute with methanol to obtain reference solutions with concentrations of 10 μg / mL, 100 μg / mL, 500 μg / mL, and 1000 μg / mL. Take 10 μL of each solution sequentially, record the peak area, and plot a standard curve, as shown in Figure 2.

[0078] 3) Chromatographic conditions

[0079] The aqueous solution containing KY386 was centrifuged at high speed (13,000 rpm for 30 minutes). The supernatant was collected and filtered through a 0.22 μm PVDF filter. The resulting aqueous solution was mixed with chromatographically pure methanol at a 1:1 ratio and then subjected to quantitative analysis by liquid chromatography. The specific chromatographic conditions were as follows: octadecyl bonded silica gel was used as the packing material (Phenomenex Gemini C18, 4.6 × 250 mm, 5 μm or equivalent column); 0.1% formic acid was used as mobile phase A, and methanol was used as mobile phase B, with gradient elution according to the table below; the column temperature was 30 °C; the detection wavelength was 246 nm; the flow rate was 1.0 mL per minute; and the injection volume was 10 μL.

[0080] Table 2: Chromatographic Analysis Conditions

[0081] 1.3 Solubility Test

[0082] Take an appropriate amount of KY386 and place it in a suitable container containing 40 mL of the above solution (20 mL for organic solvents) until an excess of KY386 is present in the solution. Place the sample in a constant temperature shaking incubator, maintain the temperature at 25°C, and shake at a suitable speed (100 rpm). Take samples at each predetermined time point according to the specifications in Table 1, check the pH, and then centrifuge (10000 rpm, 10 minutes). Dilute with the appropriate solution (to dissolve the API for solubility studies) to the appropriate concentration for HPLC analysis (Note: accurately record the dilution ratio after the experiment) to obtain solubility data.

[0083] Table 3: Water solubility analysis of KY386

[0084] The above experimental data proves that KY386 has poor solubility in aqueous solutions at different pH values, and is a nearly insoluble or insoluble compound.

[0085] 1.4 Solubility Analysis of KY386 in Common Organic Solvents

[0086] For each solvent in the list of commonly used organic solvents in Table 4, 2 mg of KY386 was weighed out. First, 50 μL of each solvent listed in the table was added to promote dissolution using methods such as vortexing and sonication. If the solvent dissolved completely, the approximate solubility was greater than 40 mg / mL. If it did not dissolve completely, more solvent was added to 100 μL to promote dissolution. If the solvent dissolved completely, the solubility was greater than 20 mg / mL. Otherwise, the next volume was added, and so on, adding 200 μL, 400 μL, 1000 μL, and 2000 μL of solvent. After each volume dissolved completely, an approximate solubility was calculated until a certain volume was reached where the solvent could not be dissolved completely. The approximate solubility range was recorded. The results are shown in Table 4.

[0087] Table 4: Solubility results of KY386 in different solvents

[0088] 1.5 Solubility Analysis of KY386 in Raw Materials and Auxiliary Materials

[0089] Take an appropriate amount of KY386 and add it to PEG400, PEG300, Tween 80, and polyoxyethylene (35) castor oil ELP respectively to investigate the solubility of KY386 raw material in each solvent at different temperatures.

[0090] Table 5: Solubility analysis results of KY386 in raw materials and auxiliary materials

[0091] Results analysis: The solubility of KY386 active pharmaceutical ingredient in PEG400 at room temperature was 38.0 mg / mL, in PEG300 it was 38.3 mg / mL, in Tween 80 it was 35.2 mg / mL, and in polyoxyethylene (35) castor oil ELP it was 19.5 mg / mL. At low temperature, there was no significant difference in the solubility of the active pharmaceutical ingredient compared with room temperature.

[0092] 1.6 Solubility Analysis of KY386 Injectable Formulation in Aqueous Solution

[0093] Each vial of KY386 injection contains 1.4 mL of PEG300, 1.36 mL of injection-grade Tween 80, 0.7 mL of polyoxyethylene (35) castor oil ELP, and 50 mg of KY386. Take one vial of KY386 injection preparation, draw 1 mL of the preparation with a sterile syringe, place it in a clean container, and dilute it with sterile water at ratios of 1:5, 1:10, and 1:50, respectively. After standing for one hour, take a sample, then centrifuge (10,000 rpm, 10 minutes), and dilute it with the corresponding solution (to dissolve the API for solubility studies) to the appropriate concentration for HPLC analysis (Note: accurately record the dilution ratio after the experiment) to obtain solubility data.

[0094] The following data shows the solubility of KY386 in various diluted aqueous solutions:

[0095] 2. Compatibility of raw and auxiliary materials

[0096] The compatibility assessment of raw materials and excipients for KY386 injection was conducted by testing four different mixtures under high temperature (60°C) and light irradiation (5500 lx ± 500). The composition of the four mixtures is shown in Table 6.

[0097] Table 6: Test Design for Compatibility of Raw Materials and Auxiliary Materials

[0098] Table 7: Results of compatibility tests of raw materials and auxiliary materials, where impurities are presented as percentages, such as 0.039, which means that the total amount of the impurity is 0.039%.

[0099] All samples taken at 5, 10, and 30 days were visually observed and analyzed by HPLC. The results of the properties and related substance compatibility tests are shown in Table 7.

[0100] The results showed that all raw material and excipient compatibility samples were yellow viscous liquids at the initial time point, and remained unchanged for 30 days under high temperature and 10 days under light. The solution color remained between yellow colorimetric solutions 6 and 8 under different initial time points. The color slightly deepened under the influencing factor test, but this deepening was unrelated to the addition of raw materials and excipients.

[0101] By comparing the results of related substances in 4 samples, it was found that after adding Tween 80 (Gedian Renfu Pharmaceutical Excipients Co., Ltd.), new impurities with an RRT of 0.821 could be detected under both high temperature and light conditions, and the impurities exceeded the standard (0.2%) at the initial time point. However, after adding polyoxyethylene (35) castor oil, no significant non-specific impurities were detected.

[0102] For sample 3 (KY386+PEG300+polyoxyethylene (35) castor oil), after 10 days of high temperature testing, the impurity RRT increased from 0.821 to 0.451%, and the impurity RRT increased from 1.650 to 0.241%. However, these two impurities were not detected at the 30-day sampling point. There may have been an error in the 10-day sampling point. To confirm the compatibility between PEG300 and polyoxyethylene (35) castor oil, a high-temperature test was performed on sample 5 (KY386+PEG300+polyoxyethylene (35) castor oil). The test results are shown in Table 8. The impurities did not increase at any sampling point.

[0103] Table 8: ELP validation data for API+PEG300+polyoxyethylene (35) castor oil

[0104] In summary, the addition of Tween 80 resulted in the generation of unknown impurities at RRT 0.821, exceeding the limit by 0.2% at day 0. The addition of polyoxyethylene (35) castor oil resulted in no significant generation of unknown impurities at day 0. PEG300 and polyoxyethylene 35 castor oil are compatible with APIs, but further research is needed on Tween 80 from different excipient manufacturers.

[0105] Table 9: Test results of Tween 80 from different manufacturers

[0106] By comparing the results of the relevant substances in Table 9, the total impurities of Tween 80 (batch number 20230110, Nanjing Well Pharmaceutical Group Co., Ltd.) increased from 0.125% to 0.147% after 30 days of high temperature; and increased from 0.125% to 0.198% after 10 days of light exposure. The total impurities increased slightly, but both were far below the limit.

[0107] Tween 80 (batch number 20230111, Gedian Renfu Pharmaceutical Excipients Co., Ltd.) showed 0.438% new impurities at day 0, and these impurities remained and increased under high temperature and light conditions. After 30 days at high temperature, the total impurities increased from 0.599% to 1.135%; after 10 days of light exposure, the impurities remained basically unchanged, with a total impurity of 0.605%.

[0108] In summary, Tween 80 (Nanjing Well Pharmaceutical Group Co., Ltd.) is compatible with the active pharmaceutical ingredient, but high-temperature storage is recommended.

[0109] 3. Determining the dosage of auxiliary materials

[0110] 3.1 Determination of the dosage of polyoxyethylene (35) castor oil

[0111] Based on the safe dosage of Tween 80, and calculated according to the density of Tween 80 (1.08), the dosage of Tween 80 is approximately 1.44 mL / vial.

[0112] The dosage of Tween 80 was fixed at 1.4 mL / vial. The effect of different amounts of polyoxyethylene (35) castor oil on insoluble microparticles in the diluted sample was investigated. The formulation information and results are shown in Table 10.

[0113] Table 10: Formulation Information and Results

[0114] Results Analysis: When the amount of Tween 80 was 1.4 mL, the insoluble particles met the standard within 1 hour when the amount of polyoxyethylene (35) castor oil was 0.625-0.7 mL, and within 2 hours when the amount of polyoxyethylene (35) castor oil was 0.7 mL. However, the insoluble particles did not meet the standard within 2 hours when the amount of polyoxyethylene (35) castor oil was less than or equal to 0.675 mL. Therefore, the amount of polyoxyethylene (35) castor oil in each vial was determined to be 0.7 mL.

[0115] 3.2 Determining the Dosage of Tween 80

[0116] With a fixed formulation volume of PEG300 (1.25 mL) and polyoxyethylene 35 castor oil (0.7 mL), the amount of Tween 80 per vial was reduced to investigate the effect of different Tween 80 dosages on the insoluble microparticles in the diluted sample. The corresponding formulation information and results are shown in Table 11.

[0117] Table 11: Formulation Information and Results

[0118] Results analysis: The amount of polyoxyethylene (35) castor oil was fixed at 0.7 mL. When the amount of Tween 80 was 1 mL, the limit was exceeded at 0 h. When the amount of Tween 80 was 1.2 mL, the insoluble particles met the standard within 1 h, but exceeded the limit at 2 h. When the amount of Tween 80 was 1.3 mL, the insoluble particles exceeded the limit at 1 h. When the amount of Tween 80 was 1.4 mL, the insoluble particles were qualified and much smaller than the limit within 2 h, and the repeatability was good. Therefore, the formulation amount of Tween 80 can be determined to be 1.4 mL.

[0119] 3.3 Determination of PEG300 dosage

[0120] Based on a liquid density of 1.08 g / mL and a formulation specification of 3.5 mL: 50 mg, the dosage of PEG300 is determined to be 1.36 mL.

[0121] 4. Development of production processes

[0122] The manufacturing process of polycyclic aromatic hydrocarbon (PAH) injections includes weighing, solution preparation, filtration, filling, and capping. The effects of solution preparation temperature, feeding sequence, nitrogen purging method, sterilization conditions, and other processes were investigated. The preparation of PAH injections is shown in Figure 1.

[0123] 4.1 Solution preparation temperature

[0124] The solubility test of the active pharmaceutical ingredient (API) showed that the API dissolves slowly at room temperature. Therefore, different solution preparation temperatures were designed to investigate the effects of solution preparation at 60℃, 40℃, and 20℃ on the dissolution time of the API and the stability of the samples.

[0125] Table 12: Summary of test results at different solution preparation temperatures

[0126] The dissolution time of the active pharmaceutical ingredient (API) at 60℃ was 15 min, at 40℃ it was 80 min, and at 20℃ it was 225 min. It can be seen that the dissolution rate of the API is significantly faster when the solution is prepared at 60℃. There are no significant differences in related substances and properties within 24 hours at the three solution preparation temperatures. Since high temperature has a greater impact on related substances, the solution preparation temperature can be determined by the changes in related substances. Based on the changes in related substances at different solution preparation temperatures, the solution preparation temperature was determined to be 60℃.

[0127] 4.2 Nitrogen purging

[0128] 4.2.1 Investigation of Nitrogen Filling Methods

[0129] The effects of headspace nitrogen purging, solution + headspace uniform nitrogen purging, and no nitrogen purging on sample stability were investigated. Table 13: Summary of experimental results for different nitrogen purging methods.

[0130] The results showed that different nitrogen purging methods had a significant impact on the related substances in the product. Nitrogen protection can increase the stability of the sample; therefore, nitrogen protection is preferably performed in the headspace and solution during preparation.

[0131] 4.2.2 Investigation of Nitrogen Filling Level

[0132] The effects of different headspace nitrogen levels with residual oxygen of 1%, 2%, and 3% on sample stability were investigated.

[0133] Table 14: Summary of Test Results for Different Nitrogen Filling Levels

[0134] The results showed that when the headspace residual oxygen content was below 3%, different headspace nitrogen levels had no effect on the relevant substances and their contents, but did affect the color of the solution. Since the solution color would deepen during high temperature and light exposure, it was preferable to control the headspace residual oxygen content below 2%.

[0135] 4.3 Screening of sterilization conditions

[0136] Based on the decision tree for sterilization process selection, three conditions (121℃ for 12 min, 121℃ for 8 min, and 115℃ for 30 min) were investigated.

[0137] Table 15: Results of Screening Related Substances for Sterilization Conditions

[0138] Table 16: Summary of Sterilization Condition Screening Stability Results

[0139] After sterilization at 121℃ for 12 minutes, 121℃ for 8 minutes, and 115℃ for 30 minutes, the relevant substances and their contents showed no significant difference compared with the unsterilized samples, indicating that the samples can withstand sterilization at 121℃ for 12 minutes.

[0140] After sterilization at 121℃ for 12 minutes, the stability of the samples after 30 days at 60℃ or 10 days under light was not significantly different from that of the unsterilized samples. This indicates that sterilization at 121℃ for 12 minutes has no effect on sample stability. The sterilization parameters are tentatively set at 121℃ for 12 minutes.

[0141] 4.4 Feeding sequence

[0142] The raw materials consisted of PEG300, Tween 80, and polyoxyethylene (35) castor oil. The three excipients were miscible. Different API addition orders may affect the API dissolution rate. Therefore, the dissolution rates of API in PEG300-API, PEG300-Tween 80-API, and PEG300-Tween 80-polyoxyethylene (35) castor oil-API were investigated.

[0143] Table 17: Summary of Data on Feeding Sequence

[0144] The results showed that different API addition orders did not significantly affect the API dissolution time. Since the heat preservation could be stopped after the raw material was completely dissolved, the preferred addition order to reduce the cooling time was PEG300-API-Tween 80-polyoxyethylene (35) castor oil.

[0145] 4.5 Packaging System

[0146] KY386 injection, with a specification of 50mg / 3.5mL, is packaged in a brown borosilicate glass vial for injection, sealed with a chlorobutyl rubber stopper for injection preparations, and sealed with an aluminum-plastic combination cap for injection preparations.

[0147] 4.6 Microbial Attributes

[0148] KY386 injection, 50 mg / 3.5 mL, used for the proposed clinical studies, is a typical sterile product. In accordance with the requirements of the 2020 edition of the Chinese Pharmacopoeia, release and monitoring for sterility and bacterial endotoxins were conducted during stability testing.

[0149] 4.7 Compatibility

[0150] KY386 injection is available in 50 mg / 3.5 mL form. It is diluted with 5% glucose injection to prepare a ready-to-use injection for intravenous infusion. The compatibility of the reconstitution solvent with the clinical delivery device was investigated. Following the proposed clinical usage method, the solution was reconstituted with 5% glucose injection and then infused. Sample information is shown in Table 18, the compatibility stability test design is shown in Table 19, and the specific compatibility study results can be found in Table 20.

[0151] Table 18: Sample Information

[0152] Table 19: Compatibility Stability Test Scheme

[0153] 4.8. Examine the insoluble microparticles, particle size, pH value, and osmolality after dilution of the formulation to determine whether the formulation composition meets the requirements for clinical application.

[0154] pH Measurement: Mettler pH meter S220, pH standard buffer solution. Take one 50mg / 3.5mL vial of KY386 injection and use at room temperature. Carefully open one vial of KY386 injection, draw all contents with a 50mL syringe, and then inject into 100mL of 5% glucose solution. Mix the sample contents, shake well, and degas under sonication for 5 minutes. Complete the process within half an hour and label it as sample solution #1. Select three or two suitable standard buffer solutions to calibrate the instrument so that the pH value of the sample solution is between them. After automatic calibration of the instrument with two standard buffer solutions, the slope should be 90%–105%, and the drift value should be within 0±30mV or ±0.5 pH units. Then verify with a third standard buffer solution with a pH between the two calibration buffer solutions and as close as possible to the sample, until the instrument reading differs from the specified value of the verification buffer by no more than ±0.05 pH units. Before each change of standard buffer solution or sample solution, thoroughly wash the electrode with purified water and then aspirate the water. Place the electrode into the sample solution prepared in section 4.1 and press the reading key to start the measurement. Adjust the electrode probe to be about 10 mm from the bottom of the container.

[0155] Insoluble particulate matter detection: Perform according to the standard procedures for a laminar flow hood and insoluble particulate matter detector (JWL-6A). Take four vials of 50mg / 3.5mL KY386 injection and use at room temperature. Wear a particle-free hood and gloves, remove the outer label from the sample vials, rinse the outer wall of the containers with pure water, and let them air dry on the laminar flow hood. Carefully open the containers, dissolve each sample vial in 100mL of 5% glucose solution, and quickly mix the sample contents. Transfer the mixture to a clean, particle-free 200mL mobile phase bottle, shake well, and degas under sonication for 5 minutes, completing the process within half an hour. The operating environment should not introduce foreign particles, and the operation before measurement should be performed on a laminar flow hood. Glassware and other necessary supplies should be clean and particle-free. Take 50mL of particle-free water for the test. The requirement is that the number of particles 10μm and above should not exceed 10 per 10mL, and the number of particles 25μm and above should not exceed 2; otherwise, the environmental blank test will fail. Rinse the system with particle-free water and perform 5 cycles. Place the prepared sample on the sampler, adjust the sampling probe to approximately 5 mm from the bottom of the container, and without stirring, directly extract an appropriate amount of liquid using the instrument (ensuring no air bubbles are drawn in). Analyze the sample according to the prescribed method. Set the software parameters according to the table below for sample testing:

[0156] Osmometry Measurement: Using the STY-1ADK osmometer, prepare four vials of 50 mg / 3.5 mL KY386 injection at room temperature. Carefully open one vial of KY386 injection, dissolve the sample in 100 mL of 5% glucose solution, mix the contents, shake well, and degas under sonication for 5 minutes. Complete the process within half an hour and label this as Sample Solution #1 (SPL#1). Carefully open three vials of KY386 injection, dissolve the sample in 100 mL of 5% glucose solution, mix the contents, shake well, and degas under sonication for 5 minutes. Complete the process within half an hour and label this as Sample Solution #2 (SPL#2). Add 60 μL of SPL#1 to the sample tube, place it on the measuring head, lower it into the cooling bath, and start the instrument for detection. Repeat the measurement twice, re-sample each time. Add 60 μL of SPL#2 to the sample tube, place it on the measuring head, lower it into the cooling bath, and start the instrument for detection. Repeat the measurement twice, re-sample each time. After each test, wipe the frost off the sensor with absorbent paper before performing the next test. Use a clean test tube for each repeated sample test to avoid affecting the consistency of the results.

[0157] KY386 Content Analysis: System Suitability: Blank (Diluent) Check the blank chromatogram from the last injection. In the blank solution chromatogram, the retention time of KY386 should not show any interfering peaks. If interference is present, the interference value must not exceed the effective reaction value in the sensitivity solution.

[0158] The signal-to-noise ratio (S / N) of the KY386 peak in the standard solution sensitivity solution should not be less than 10.

[0159] The resolution between the main peak KY386 and its adjacent peaks in the sample solution should be no less than 1.5.

[0160] The RSD% values ​​of the KY386 peak area in the five injections of standard solutions were all less than 2.0% for the system.

[0161] The recovery rate of the inspection standard STD#2 should be between 98.0% and 102.0%.

[0162] Standard recovery should be maintained throughout the entire operation, with all standard recoveries (within parentheses) between 98.0% and 102.0%. Calculate the recovery of the reference standard as follows: WSTD#1 = Weight (mg) of KY386 standard in Reference Standard Solution #1 (STD#1) WSTD#2 = Weight (mg) of KY386 standard in Reference Standard Solution #2 (STD#2) ASTD#1 = Average peak area of ​​KY386 in five initial injections of Standard Solution #1 (STD#1) ASTD#2 = Peak area of ​​KY386 in Standard Solution 2 (STD#2)

[0163] Standard recovery rate recovery A STD#1 =The average peak area of ​​the main peak in the first five injections of STD#1. A BracketingSTD#1 = Peak area of ​​the main peak.

[0164] The recovery rate of the standard injection solution was calculated using the following method:

[0165] Use the formula below to calculate the test results for each sample solution injection, expressed as a percentage as required on the label.

[0166] Au = the peak area of ​​KY386 in the sample solution used for measurement.

[0167] As = the average peak area of ​​KY386 in the initial 5 injections of STD#1 used for determination.

[0168] Ws = Weight (mg) of KY386 in STD#1 used for determination.

[0169] The valence of P=KY386 standard

[0170] Du = Sample dilution factor

[0171] Ds = Standard dilution factor

[0172] Wu = Please modify the weight in the sample solution to be tested, by multiplying the sample volume (mL) by the specification (50mg / 3.5mL).

[0173] ρ = sample density (g / cm³) 3 )

[0174] PSN = Product Specification (50mg / 3.5mL)

[0175] Insoluble microparticles, particle size determination, and osmotic pressure determination: Take 3.5 mL of the full-formulation preparation sample and dilute it into 100 mL of 5% glucose injection at different temperatures (25℃, 30℃, 35℃). Detect the insoluble microparticles and particle size at 0h, 1h, 2h, and 3h. Store the diluted sample at room temperature (25℃). Particle size determination was performed according to the standard procedure of the Malvern laser particle size analyzer. Insoluble microparticle indices were determined according to the standard procedure of the insoluble microparticle detector (JWL-6A). Osmotic pressure molar concentration was determined according to the standard procedure of the osmometer (STY-1ADK).

[0176] Table 20: Results of particle size analysis in compatibility stability tests

[0177] Table 21: Summary of Insoluble Particle Detection Results in Compatibility Stability Study

[0178] Table 22: Summary of Osmotic Pressure Test Results in Compatibility Stability Study

[0179] Table 23: Summary of pH test results

[0180] Results analysis: The insoluble particles and particle size after dilution are closely related to the dilution temperature. After dilution at 25℃ and standing at room temperature for 3 hours, 2% of the particles with a diameter of 5133nm were generated. After dilution at 25℃ and standing at room temperature for 3 hours, the insoluble particles exceeded the limit. At dilution temperatures of 25℃-35℃, the particle size was between 11nm and 13nm within 2 hours, and the insoluble particles were all within the standard range.

[0181] After formulation, the osmotic pressure molar concentration is close to that of an isotonic solution, and the osmotic pressure remains stable within 3 hours.

[0182] The pH value of the formulation after mixing is 4.90, which is slightly higher than that of 5% glucose injection (4.70), and there is no risk of clinical use.

[0183] Since this product is used clinically by diluting it in 100mL of 5% glucose injection, which can meet clinical needs within 2 hours, the clinical dilution temperature needs to be controlled at 25℃-35℃. The formulation is reasonable and can meet clinical needs.

[0184] 5. Stability data of the injectable formulation

[0185] The following stability tests of polycyclic aromatic hydrocarbon injections were conducted in accordance with the requirements of the 2020 edition of the Chinese Pharmacopoeia.

[0186] 5.1 Test Samples

[0187] Table 24: Sample Information

[0188] 5.2 Influencing Factors Experiment

[0189] Influencing factor tests were conducted on non-GMP batches of KY386 injection to assess possible degradation pathways and evaluate the suitability of packaging materials and storage conditions.

[0190] The batch information and sampling points of the influencing factor test are summarized in Table 25.

[0191] Table 25: Test Protocol for Factors Affecting KY386 Injection

[0192] Remark:

[0193] 1. T = Properties, clarity and color, related substances, content determination, visible foreign matter and insoluble particulate matter test;

[0194] 2. / = Not detected;

[0195] 3. A = Osmotic molar concentration.

[0196] 5.3 Stability Study

[0197] Table 26: Stability study protocol for KY386 injection, 50 mg / 3.5 mL (Non-GMP batch)

[0198] Remark:

[0199] 1. T = Properties, pH value, clarity and color, related substances, osmolality, moisture, viscosity, content, visible foreign matter and insoluble particles;

[0200] 2. / = Not detected

[0201] Table 27: Stability study protocol for KY386 injection, 50 mg / 3.5 mL (GMP batch)

[0202] Remark:

[0203] 1. A = Properties, content, color and clarity, osmolality, insoluble particles, pH value, related substances, viscosity, visible foreign matter and moisture detection.

[0204] 2.C = Container sealing test

[0205] 3. M = Bacterial endotoxin and sterility test

[0206] 4. / = Not detected

[0207] 5.4 Stability Data

[0208] 5.4.1 Influencing Factor Experiment

[0209] Table 28: Influencing Factors Test - High Temperature Test (Batch No.: 20230110)

[0210] Table 29: Influencing Factors Test - Light Irradiation Test (Batch No.: 20230110)

[0211] Table 30: Influencing Factors Test - Freeze-Thaw Test (Batch No.: 20230110)

[0212] 5.4.2 Accelerated Testing

[0213] Table 31: Accelerated test data at 40℃±2℃ / 75±5%RH (Batch No.: 20230110)

[0214] Table 32: Accelerated test data at 25℃±2℃ / 60±5%RH (Batch No.: CT23E0015)

[0215] 5.4.3 Long-term test

[0216] Table 33: Accelerated test data at 30℃±2℃ / 65±5%RH (Batch No.: 20230110)

[0217] Table 34: Long-term test data at 5℃±3℃ (batch number: CT23E0015)

[0218] 5.4.4 Compatibility Stability Test

[0219] Table 35: Compatibility Stability Test Data (Batch No.: 20230110)

[0220] Table 36: Compatibility Stability Data (Batch No.: CT23E0015)

Claims

1. A polycyclic compound injection, characterized in that, The injection contains a polycyclic compound of formula I, low-viscosity polyethylene glycol, Tween 80, and polyoxyethylene (35) castor oil, wherein the concentration of the polycyclic compound of formula I is 0.1-15 mg / mL.

2. The injectable preparation according to claim 1, characterized in that, The injection contains approximately 14.28 mg / mL of the polycyclic compound.

3. The injectable preparation according to claim 1, characterized in that, The volume ratio or mass ratio of the low-viscosity polyethylene glycol, Tween 80, and polyoxyethylene (35) castor oil is approximately 2:2:

1.

4. The injectable preparation according to claim 3, characterized in that, The low-viscosity polyethylene glycol is PEG300, PEG400, or a mixture thereof.

5. The injectable preparation according to claim 1, characterized in that, The injection is water-free or its water content is controlled within 1% by mass.

6. A method for preparing the polycyclic compound injection according to any one of claims 1-5, characterized in that, Includes the following steps: S1. After heating the low-viscosity polyethylene glycol to 60±5℃, keep the temperature constant and keep it under nitrogen (0.03Mpa) for at least 20 minutes; S2. Under stirring conditions, add the polycyclic compound and stir until fully dissolved; S3. Add Tween 80, maintain nitrogen pressure (0.03 MPa) and stir thoroughly for 10 minutes; and S4. Add polyoxyethylene (35) castor oil, continue stirring and nitrogen purging for 30 minutes until the mixture is homogeneous, thus obtaining the polycyclic compound injection.

7. The method according to claim 6, characterized in that, The volume ratio or mass ratio of the low-viscosity polyethylene glycol, Tween 80, and polyoxyethylene (35) castor oil is approximately 2:2:1, and the concentration of the polycyclic compound is 0.1-15 mg / mL. Preferably, the low-viscosity polyethylene glycol is PEG300, PEG400, or a mixture thereof.

8. A ready-to-use polycyclic aromatic hydrocarbon injection, characterized in that... The injection contains water, a polycyclic compound injection according to any one of claims 1-5, and an osmotic pressure regulator, wherein the ready-to-use polycyclic compound injection has a pH of 3.2-6.5 and is isotonic, and preferably the concentration of the polycyclic compound is 0.1-0.5 mg / mL.

9. A method for analyzing and separating the content and impurities of the polycyclic compound of Formula I in the injectable formulation of the polycyclic compound according to any one of claims 1-5, characterized in that, The chromatographic conditions were as follows: Octadecyl-bonded silica gel was used as the stationary phase (Phenomenex Gemini C18, 4.6 × 250 mm, 5 μm or equivalent column); 0.1% formic acid was used as mobile phase A, and methanol as mobile phase B, with gradient elution performed according to the table below; the column temperature was 30℃; the detection wavelength was 246 nm; the flow rate was 1.0 mL per minute; the injection volume was 10 μL; and the gradient elution time was set to 37 minutes. Chromatographic analysis conditions

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