Polycyclic compound oral preparation and preparation method thereof

Abstract

The invention belongs to the field of cancer medicine preparations, and discloses a polycyclic compound oral preparation and a preparation method thereof. The polycyclic compound oral preparation contains a polycyclic compound as shown in a formula I, polyethylene glycol, propylene glycol monooctoate and polyoxyethylene (40) hydrogenated castor oil, wherein the concentration of the polycyclic compound as shown in the formula I is 0.1-80 mg / mL. According to the polycyclic compound oral preparation disclosed by the invention, the solubility of the polycyclic compound shown in the formula I in an aqueous solution is improved, the preparation can obtain a self-emulsifying particle size smaller than 100nm in the aqueous solution, and meanwhile, the polycyclic compound oral preparation has good stability, dissolution rate, oral bioavailability, body tolerance or safety of auxiliary materials, low cost, easiness in production and storage resistance, # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of cancer drug preparations, and in particular relates to an oral preparation of a polycyclic compound and a preparation method thereof. Background Art

[0002] DHX33 belongs to the RNA helicase protein family that contains the DEAD / H box. DEAD / H stands for the amino acid abbreviation Asp-Glu-Ala-Asp / His. This sequence, along with several other conserved amino acid sequences, is found in the protein sequences of RNA helicase family members and is highly involved in nucleic acid substrate binding and ATP hydrolysis. While these family members share these common sequences, each RNA helicase has unique specificity and biological functions. The human DHX33 protein has a molecular weight of 72 kDa and functions to unwind nucleic acids. It utilizes the bioenergy released by ATP hydrolysis to drive conformational changes in RNA and protein complexes, thereby participating in a variety of RNA metabolic activities, specifically, RNA transcription, splicing, editing, translation, and degradation. DHX33's functions are not limited to RNA modification. Research has shown that in addition to unwinding RNA duplexes, DHX33 also participates in DNA metabolism. Specifically, DHX33 can unwind the double-stranded structure of DNA and plays a vital role in gene expression. In an in vitro enzyme reaction system, it was also found that DHX33 can unwind the hybrid double-stranded structure of DNA / RNA.

[0003] Studies have shown that DHX33 binds to the promoters of various cancer-related genes, affecting the methylation status of DNA. This in turn regulates the expression of multiple cancer genes and signaling pathways associated with tumor development at the genomic level, playing 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 crucial cell growth regulator, DHX33 is highly expressed in various cancers. The development and progression of many cancers depend on the high expression of the DHX33 protein. Genetic knockout of the DHX33 gene can significantly inhibit the development and progression of lung cancer driven by the RAS oncogene. In vivo and in vitro experiments have confirmed that inhibition of the DHX33 protein significantly suppresses the development and progression of various cancers, including breast cancer, colon cancer, glioma, and lymphoma.

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

[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 a 24-hour equilibrium solubility in water of less than 0.01 mg / mL, making it a poorly soluble compound. Currently, there are no reports on oral preparations of KY386. How to improve the water solubility of the inhibitor and develop an oral preparation with various properties that meet the needs of clinical trials, including stability, ease of production, low formulation cost, high body tolerance, and ease of transportation and storage, is an urgent problem that needs to be solved. Summary of the Invention

[0006] In view of this, the purpose of the present invention is to provide an oral preparation of polycyclic compounds and a preparation method to solve the problem of poor water solubility of polycyclic compounds in the prior art, while providing a stable oral preparation with high body tolerance.

[0007] In one aspect, the present invention provides an oral preparation of a polycyclic compound, which comprises a polycyclic compound of formula I, polyethylene glycol, propylene glycol monocaprylate, and polyoxyethylene (40) hydrogenated castor oil.

[0008]

[0009] In an embodiment of the first aspect of the present invention, the concentration of the polycyclic compound of formula I in the oral formulation is about 0.1-80 mg / mL.

[0010] In an embodiment of the first aspect of the present invention, the polyethylene glycol may be PEG300, PEG400, PEG4000 or PEG6000 or a mixture thereof.

[0011] In a second aspect, the present invention provides a method for preparing an oral formulation of a polycyclic compound of Formula I, comprising the following steps:

[0012] S1. Heat polyethylene glycol to 60 ± 5°C, maintain constant temperature, and maintain nitrogen bubbling (0.03 MPa) for at least 20 minutes;

[0013] S2. Add polyoxyethylene (40) hydrogenated castor oil under stirring, continue heating to 60 ± 5 ° C, and stir until fully dissolved;

[0014] S3. Add the polycyclic compound, maintain nitrogen bubbling (0.03Mpa) and stir thoroughly for 60-120 minutes until the compound is dissolved, and stop heating;

[0015] S4. Add propylene glycol monocaprylate, continue stirring and nitrogen aeration for 30 minutes until the mixture is uniform, and obtain the oral preparation of the polycyclic compound.

[0016] In an embodiment of the second aspect of the present invention, the concentration of the polycyclic compound of formula I in the oral formulation is about 0.1-80 mg / mL.

[0017] In an embodiment of the second aspect of the present invention, the polyethylene glycol may be PEG300, PEG400, or a mixture thereof; or PEG4000, PEG6000, or a mixture thereof.

[0018] In an embodiment of the present invention, the water content in the oral preparation is within 5% based on the total mass of the oral preparation.

[0019] The oral formulation of the present invention improves the solubility of the polycyclic compound represented by Formula I in aqueous solution, and the formulation can obtain a self-emulsifying particle size of less than 100 nm in aqueous solution. At the same time, it has good formulation stability, dissolution, oral bioavailability, body tolerance or safety of excipients, low cost, easy production and storage stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The chromatographic standard curve of the polycyclic compound represented by Formula I is shown, wherein the ordinate is the peak area and the abscissa is the concentration (μg / mL).

[0021] Figure 2 It was shown that the oral preparation of the present invention maintained stable dissolution in an acidic aqueous solution for 12 hours.

[0022] Figure 3 The graph shows the particle size analysis effect of the oral preparation of the present invention in aqueous solution.

[0023] Figure 4 The dissolution curve of the oral preparation of the present invention in simulated gastric fluid is shown. DETAILED DESCRIPTION

[0024] The technical solutions of the present invention will be clearly and completely described below in conjunction with specific embodiments. However, it should be understood by those skilled in the art that the embodiments described below are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work are within the scope of protection of the present invention.

[0025] In one aspect, the present invention provides an oral preparation of a polycyclic compound, which comprises a polycyclic compound of formula I, polyethylene glycol, propylene glycol monocaprylate, and polyoxyethylene hydrogenated (40) castor oil.

[0026]

[0027] In an embodiment of the present invention, the concentration of the polycyclic compound of Formula I in the oral formulation is 0.1-80 mg / mL. In a specific embodiment, the concentration of the polycyclic compound of Formula I in the oral formulation can be about 0.2 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 2.0 mg / mL, 3.0 mg / mL, 4.0 mg / mL, 5.0 mg / mL, 10.0 mg / mL, 20.0 mg / mL, 30.0 mg / mL, 40.0 mg / mL, 50.0 mg / mL, 60.0 mg / mL, 70.0 mg / mL, 80.0 mg / mL and values ​​between any two of the above values, for example, about 0.25 mg / mL, 0.35 mg / mL.

[0028] In an embodiment of the present invention, the polyethylene glycol may be PEG300, PEG400, or a mixture thereof, or PEG4000, PEG6000, or a mixture thereof.

[0029] In the oral preparation of the polycyclic compound of the present invention, the volume ratio of polyethylene glycol can be about 9-81%, based on the total volume of the oral preparation, for example, about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, and values ​​between any two of the above values, for example, about 12.5%, 20.5%, 32.5%, 42.5%, 52.5%, etc.

[0030] In the oral preparation of the polycyclic compound of the present invention, the volume ratio of propylene glycol monocaprylate can be about 10-30%, based on the total volume of the oral preparation, for example, about 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, and values ​​between any two of the above values, for example, about 16.5%, 18.5%, 21.5%, 24.5%, 26.5%, etc.

[0031] In the oral preparation of the polycyclic compound of the present invention, the volume ratio of polyoxyethylene (40) hydrogenated castor oil can be about 9-81%, based on the total volume of the oral preparation, for example, about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, and values ​​between any two of the above values, for example, about 12.5%, 20.5%, 32.5%, 42.5%, 52.5%, etc.

[0032] In an embodiment of the present invention, the oral formulation may further contain an antioxidant. In a specific embodiment, the mass ratio of the antioxidant may be about 0.02-0.1%, such as about 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, and values ​​between any two of the above values, based on the total mass of the oral formulation.

[0033] In an embodiment of the present invention, the water content of the oral preparation is controlled within 5% based on the total weight of the oral preparation. Obviously, the water content needs to be controlled during the preparation process to prevent excessive water content from causing compound instability.

[0034] Obviously, in order to further enhance the stability of the oral formulation of the polycyclic compound, other substances that are harmless and do not react with the polycyclic compound of Formula I can be added. These substances are generally:

[0035] Protective gas, by evacuating the polycyclic compound oral preparation to greatly reduce the content of gases with certain activity such as O2 in the oral preparation, and then filling it with an inert gas that does not react with the polycyclic compound shown in Formula I, polyethylene glycol, propylene glycol monocaprylate or polyoxyethylene (40) hydrogenated castor oil, such as N2 or other inert gases. These protective gases will protect the stability of the preparation and prevent oxidation reaction; or

[0036] Antioxidants, by adding appropriate amounts of antioxidants, such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) or vitamin E, to the oral preparation of polycyclic compounds, further improve their stability.

[0037] In an embodiment of the present invention, in order to improve patient compliance during use, the oral preparation of the present invention may further contain an anesthetic or analgesic drug at a dose permitted by the pharmacopoeia or formulary.

[0038] In an embodiment of the present invention, the oral formulation of a polycyclic compound may further contain another therapeutic agent to form a compound formulation, thereby exerting a synergistic and enhanced therapeutic effect. In particular, these formulations are recommended as drugs that regulate the expression level of the DHX33 enzyme or the corresponding gene.

[0039] In some cases, in order to cope with special environments, the oral polycyclic compound preparation of the present invention may also contain antibacterial agents, antifungal agents, etc.

[0040] Obviously, the added amount of the above-mentioned protective gas, antioxidant, anesthetic or analgesic drugs, antibacterial agents, antifungal agents, etc. should not affect the solubility and other properties of the polycyclic compound represented by Formula I or enhance its stability; while the content of another therapeutic agent, a substance for regulating water-salt, electrolyte and acid-base balance, osmotic pressure, and a substance for regulating pH needs to be determined according to the purpose, prescription amount, pharmacopoeia, formulary, etc. and can be determined according to actual conditions.

[0041] Obviously, the polyethylene glycol and polyoxyethylene (40) hydrogenated castor oil defined in the present invention should each be liquid at room temperature and differential pressure, or the mixed solvent after mixing should be liquid. Both polyoxyethylene (40) hydrogenated castor oil and propylene glycol monocaprylate in the polycyclic compound oral formulation have relatively high viscosities, resulting in a relatively high viscosity of the oral formulation. From another perspective, these substances can also hinder the movement of the active ingredient molecules, thereby enhancing the stability of the polycyclic compound oral formulation.

[0042] As a specific embodiment, the unit size of the polycyclic compound oral preparation can be fixed within 0.1 mL-1.0 mL, which is convenient for oral administration and can be packaged in vials. The oral preparation can be filled into capsules and sealed. The capsule size is not limited, for example, it can be #5, #4, #3, #2, #1, or #0 capsules. The capsule material is not limited, and can be selected from liquid-filled hard capsules, liquid soft capsules, gelatin capsules, plant fiber capsules, etc.

[0043] In an embodiment of the present invention, the oral preparation can be poured into a 10 mL medium borosilicate glass tube (brown) injection bottle, which is then filled with inert protective gas, capped with a rubber stopper and sealed with an aluminum-plastic composite cap for antibiotic bottles.

[0044] The oral preparation can also be prepared as liquid-filled hard capsules or liquid soft capsules, and the packaging can be a combination of high-density polyethylene bottles or double aluminum blister packaging. Antioxidants can be added to the container holding the capsules to maintain the stability of the preparation.

[0045] In a second aspect, the present invention provides a method for preparing the above-mentioned oral formulation, comprising the following steps:

[0046] S1. Heat polyethylene glycol to 60 ± 5°C, maintain constant temperature, and maintain nitrogen bubbling (0.03 MPa) for at least 20 minutes;

[0047] S2. Add polyoxyethylene (40) hydrogenated castor oil under stirring, continue heating to 60 ± 5 ° C, and stir until fully dissolved;

[0048] S3. Add the polycyclic compound represented by Formula I, maintain nitrogen bubbling (0.03Mpa) and stir thoroughly for 60-120 minutes until the compound is dissolved, and stop heating;

[0049] S4. Add propylene glycol monocaprylate, continue stirring and nitrogen aeration for 30 minutes until the mixture is uniform, and obtain the oral preparation.

[0050] In an embodiment of the second aspect, the polyethylene glycol is PEG300, PEG400, or a mixture thereof, or PEG4000, PEG6000, or a mixture thereof.

[0051] In an embodiment of the second aspect, based on the total volume of the oral formulation, the amount of polyethylene glycol added can be about 9-81% by volume, for example, about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, and values ​​between any two of the above values, for example, about 12.5%, 20.5%, 32.5%, 42.5%, 52.5%, etc.

[0052] In an embodiment of the second aspect, based on the total volume of the oral formulation, the added amount of propylene glycol monocaprylate can be about 10-30% by volume, for example, about 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, and values ​​between any two of the above values, for example, about 16.5%, 18.5%, 21.5%, 24.5%, 26.5%, etc.

[0053] In an embodiment of the second aspect, based on the total volume of the oral formulation, the added amount of polyoxyethylene (40) hydrogenated castor oil can be about 9-81% by volume, for example, about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, and values ​​between any two of the above values, for example, about 12.5%, 20.5%, 32.5%, 42.5%, 52.5%, etc.

[0054] In an embodiment of the second aspect, an antioxidant is further added in step S3. Antioxidants are well known to those skilled in the art and may be, for example, BHA, BHT, or vitamin E. In an embodiment of the present invention, the amount of antioxidant added may be about 0.02-0.1% by mass, based on the total mass of the oral formulation, for example, about 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, and values ​​between any two of the above values.

[0055] In an embodiment of the second aspect, the amount of the polycyclic compound of Formula I added can be about 5% by mass based on the total mass of the oral formulation. In an embodiment of the present invention, the amount of the compound of Formula I added can be about 1-8% by mass, for example, about 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, and values ​​between any two of the above values.

[0056] In an embodiment of the present invention, the mass ratio and volume ratio are based on the total mass and total volume of the oral formulation of the present invention, respectively.

[0057] In this specification, the term "about" typically means + / - 5% of the stated value, more typically + / - 4% of the stated value, more typically + / - 3% of the stated value, more typically + / - 2% of the stated value, even more typically + / - 1% of the stated value, and even more typically + / - 0.5% of the stated value.

[0058] In this specification, the polycyclic compound represented by Formula I is also referred to as KY386, API or bulk drug.

[0059] The preferred embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that the following examples 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 may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention, and all such modifications and substitutions fall within the scope of the claims of the present invention.

[0060] 1. Analysis of substances related to the active ingredient KY386 in oral preparations

[0061] Instruments and reagents:

[0062] Agilent 1260 HPLC with UV detector or equivalent, model: 1260 Infinity II

[0063] HPLC column: Phenomenex Gemini 5μm*250mm*4.6mm or equivalent, P / N: 00G-4435-E0

[0064] Analytical balance with weighing accuracy of at least 0.01mg or 0.001mg, model: XP6

[0065] Dissolution Apparatus, Model DS-1206AT

[0066] Centrifuge, model Centrifuge-5424-R

[0067] Water Purifier, Model: Classic U

[0068] Grade A glassware

[0069] Class A volumetric flasks

[0070] Pure water, HPLC grade or equivalent

[0071] Methanol (MeOH), HPLC grade or equivalent

[0072] Formic acid (FA), HPLC grade or equivalent

[0073] Ethanol (EtOH), HPLC grade or equivalent

[0074] Chromatographic conditions:

[0075]

[0076]

[0077] A brief calculation method for impurities:

[0078]

[0079] Ax = peak area of ​​a single impurity (each peak area is not less than 0.05%)

[0080] As = peak area of ​​KY386 peak in 1% sample solution, or

[0081] Peak area of ​​KY386 peak in 1% reference solution

[0082] 2. Dissolution Analysis of Preparations

[0083] Instruments and reagents:

[0084] Agilent 1260 HPLC with UV detector or equivalent, model: 1260 Infinity II

[0085] HPLC column: Phenomenex Gemini 5μm*250mm*4.6mm or equivalent, P / N: 00G-4435-E0

[0086] Analytical balance with weighing accuracy of at least 0.01mg or 0.001mg, model: XP6

[0087] Dissolution Apparatus, Model DS-1206AT

[0088] Centrifuge, model Centrifuge-5424-R

[0089] Water Purifier, Model: Classic U

[0090] Grade A glassware

[0091] Class A volumetric flasks

[0092] Pure water, HPLC grade or equivalent

[0093] Methanol (MeOH), HPLC grade or equivalent

[0094] Formic acid (FA), HPLC grade or equivalent

[0095] Ethanol (EtOH), HPLC grade or equivalent

[0096] Hydrochloric acid (HCl) AR grade or equivalent

[0097] Chromatographic conditions:

[0098]

[0099] Dissolution conditions:

[0100] condition parameter Dissolution method Paddle method (sinking basket) Dissolution speed 75rpm Dissolution medium pH 1.6 buffer or simulated gastric fluid of the Chinese Pharmacopoeia Dissolution volume 900mL Dissolution temperature 37℃ Runtime 30min Flow rate fluid volume / fluid replacement volume 10mL / 10mL

[0101] Dissolution: Take 6 portions of the oral preparation of unit specification and place them in dissolution cups. Determine the dissolution according to the dissolution conditions. Take the dissolution at the sampling point, filter it with a 0.8μm water filter, discard 3mL, and take the filtrate.

[0102] Test solution: Place 5 mL of the dissolution solution in a 10 mL volumetric flask, dilute to the mark with ethanol, shake well, filter the supernatant through 0.22 μm nylon 6, discard 1 mL, and prepare a solution containing approximately 14 μg of KY386 compound per 1 mL. The remaining procedures were carried out in accordance with the Chinese Pharmacopoeia (2020 edition).

[0103] 3. Self-emulsifying particle size analysis of the preparation

[0104] Instruments and reagents:

[0105] Malvern Panalytical, zeta size radvance (Shenzhen Bay Laboratory)

[0106] Electronic balance (Model: ME104, Mettler-Toledo Instruments (Shanghai) Co., Ltd.)

[0107] pH meter (Model: S220, Mettler-Toledo Instruments (Shanghai) Co., Ltd.)

[0108] Anhydrous methanol (Batch number: C16305430)

[0109] Concentrated hydrochloric acid (37%) (Batch number: 20240606)

[0110] Ultrapure water

[0111] Detection method:

[0112] 1) Preparation of pH 3.0 aqueous solution

[0113] Add 999.92 (~1000) mL of ultrapure water to a 1 L clean glass container.

[0114] 0.08 mL of analytical grade 38% concentrated hydrochloric acid solution was added, the mixture was stirred thoroughly, and the pH was measured to adjust to 3.0.

[0115] 2) Capsule dosage

[0116] Randomly select 20 units of oral preparations, remove the packaging, and place the contents in a clean container.

[0117] Place the oral preparation in a clean, clear glass container.

[0118] Use a balance to accurately weigh 1.0 g of the oral preparation.

[0119] 3) Dilution of preparation

[0120] 999 mL of an aqueous solution with a pH of 3.0 was measured, a stirring bar was added, and the stirring speed was adjusted to 1000 rpm.

[0121] 1.0 g of the oral preparation was added to the above aqueous solution and stirred continuously to form a self-emulsifying system.

[0122] 4) Particle size test

[0123] Measure 1.0 mL of the above preparation solution and measure it three times in parallel.

[0124] The particle size of the sample was measured according to the standard operating procedure of the Malvern particle size analyzer and the particle size data was recorded.

[0125] Repeat the above steps and calibrate at least 3 times. If the calibration result is not repeatable, calibrate twice more and delete the result with larger error in the calibration record.

[0126] Example

[0127] The preferred embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that the following examples 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 may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention, and all such modifications and substitutions fall within the scope of the claims of the present invention.

[0128] In the following examples, the solubility of the polycyclic compound represented by Formula I (also known as KY386, API, or bulk drug) was analyzed with reference to the Chinese Pharmacopoeia (2020 edition). Unless otherwise specified, the materials and reagents used in the following examples were commercially available.

[0129] 1. Solubility study of KY386

[0130] 1.1 Buffer / Solution Preparation

[0131] Stock solutions and buffer volumes different from those specified may be used as long as the target concentration remains the same.

[0132] Table 1. Preparation methods of aqueous solutions with different pH values

[0133]

[0134] 1.2 Chromatographic determination of aqueous solubility of KY386

[0135] 1) Preparation of test solution

[0136] Accurately weigh an appropriate amount of KY386 raw material, add methanol to fully dissolve and dilute to a final concentration of 10 mg / mL, aspirate 10 μL, record the peak area, and calculate the content of KY386 in the sample.

[0137] 2) KY386 solubility linear range investigation: Accurately weigh an appropriate amount of KY386 standard (purity 99.9%, content 99.8%), dissolve it in methanol and prepare a 5000μg / mL solution. Then gradually dilute it with methanol to obtain reference solutions with concentrations of 10μg / mL, 100μg / mL, 500μg / mL, and 1000μg / mL, respectively. Pipette 10μL in sequence, record the peak area, and draw a standard curve, such as Figure 2 shown.

[0138] 3) Chromatographic conditions

[0139] 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 grade methanol in a 1:1 ratio and then quantitatively analyzed by liquid chromatography. The specific chromatographic conditions were: octadecyl bonded silica gel as the filler (phenomenex Gemini C18, 4.6×250 mm, 5 μm or equivalent column); 0.1% formic acid as mobile phase A, methanol as mobile phase B, with a gradient elution as shown in the table below; column temperature of 30°C; detection wavelength of 246 nm; flow rate of 1.0 mL / min; injection volume of 10 μL.

[0140] Table 2. Chromatographic conditions

[0141]

[0142]

[0143] 1.3 Solubility test

[0144] 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 maintained at 25°C and shake at an appropriate speed (100 rpm). Check the pH as specified in Table 1. Samples are taken at each predetermined time point and then centrifuged (10,000 rpm for 10 minutes). Add the appropriate solution (to dissolve the API for solubility studies) and dilute to the appropriate concentration for HPLC analysis (Note: Accurately record the dilution ratio after the experiment) to obtain solubility data.

[0145] Table 3. Water solubility analysis of KY386

[0146]

[0147] The above experimental data prove that KY386 has poor solubility in aqueous solutions of different pH values ​​and is a nearly insoluble or insoluble compound.

[0148] 1.4 Analysis of the solubility of KY386 in common organic solvents

[0149] As a highly lipophilic active pharmaceutical ingredient, KY386 has extremely low solubility in aqueous solutions, making it a poorly soluble compound. Table 4 analyzes the solubility of KY386 in several commonly used organic solvents. For each solvent listed in Table 4, 2 mg of KY386 was weighed and initially dissolved in 50 μL of each solvent listed below. Dissolution was then enhanced by vortexing, sonication, or other methods. If the solution was clear, the rough solubility was greater than 40 mg / mL. If the solution was not clear, the solvent was added to 100 μL to promote dissolution. If so, the solubility was greater than 20 mg / mL. Otherwise, the next volume was added, followed by 200 μL, 400 μL, 1000 μL, and 2000 μL of solvent. After each volume was dissolved, a rough solubility was calculated, and the solubility range was recorded until the solution was no longer clear at a certain volume. The results are shown in Table 4.

[0150] Table 4. Solubility results of KY386 in different solvents

[0151] Solvent name Solubility (mg / mL) Solvent name Solubility (mg / mL) Methanol 3.6<S<5.5 Acetonitrile 2.0<S<5.2 ethanol 2.1<S<5.4 dichloromethane 19.3<S<38.6 Isopropyl alcohol 1.0<S<2.1 Tetrahydrofuran S>40.2 acetone 9.4<S<18.8 2-Methyltetrahydrofuran 10.1<S<20.2 Ethyl acetate 5.1<S<10.4 1,4-Dioxane 18.1<S<36.2 dimethylformamide S>41.4 Dimethyl sulfoxide S>41.4

[0152] Rough solubility analysis of 1.5KY386 in aqueous solution after dissolution in oral formulation

[0153] KY386 oral formulations containing various excipient ratios were prepared to achieve KY386 concentrations of 10 mg / mL, 20 mg / mL, 40 mg / mL, 60 mg / mL, and 80 mg / mL, respectively. The concentrations of each excipient and KY386 in the oral formulations are shown in the table below. 1 mL of each KY386 oral formulation was diluted 1:10 with aqueous solutions at different pH conditions. After standing for 1 hour, the solution was observed to obtain rough solubility data.

[0154] Table 5. Approximate solubility data of KY386 with different formulations in dilute aqueous solutions:

[0155]

[0156] Note: "√" indicates complete dissolution, with a clear solution and no visible precipitation. "X" indicates incomplete dissolution, with compound precipitation in the solution. Capryol 90: Propylene glycol monocaprylate; RH40: Polyoxyethylene (40) hydrogenated castor oil

[0157] 2. Comparison of self-emulsification effects

[0158] First, the emulsifier polyoxyethylene (40) hydrogenated castor oil and the co-emulsifier polyethylene glycol were mixed in a volume ratio of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, and 9:1. The resulting mixture was called MIX. Then, the emulsifier and co-emulsifier were mixed as a whole with the oil phase propylene glycol monocaprylate in a volume ratio of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, and 9:1. 100 μL of the mixed liquid was added to 10 mL of deionized water (pH 6.8, 3, and 1.6, respectively) and stirred at 120 rpm on a magnetic stirrer at room temperature. After stabilization, the appearance grade of the emulsion was evaluated (see the table below for the emulsification grade appearance evaluation criteria).

[0159]

[0160] The mixture containing the above excipients in various proportions was diluted and the self-emulsification effect was observed. The results are shown in Table 6.

[0161] Table 6. Self-emulsification effect of pharmaceutical excipients with different ratios

[0162]

[0163]

[0164] 3. Effect of different ratios of self-emulsifying formulations on KY386 drug loading

[0165] The experimental method is briefly described as follows: Oral formulations were prepared according to the following excipient ratios. A specific amount of KY386 compound was then added in batches, with mixing at 1000 rpm. If the first addition of KY386 completely dissolved, a second appropriate amount of KY386 compound was added. These two additions were used to analyze the crude solubility of the compound in the various formulations. Dissolution, precipitation, and solution clarity were carefully recorded for each formulation.

[0166] Table 7. Effects of different proportions of propylene glycol monocaprylate in the oil phase on the drug loading of KY386 in the formulation

[0167]

[0168] Similar to the above experimental method, but with different formulation ratios, prepare oral formulations according to the following excipient ratios. Then, add a certain amount of KY386 compound in batches and mix at 1000 rpm. If the first addition of KY386 completely dissolves, add a second appropriate amount of KY386 compound. Based on these two additions, analyze the crude solubility of the compound in the different formulations. Carefully record the dissolution of the compound, compound precipitation, and solution clarity in each formulation.

[0169] Table 8. Effects of different ratios of emulsifier and co-emulsifier on the drug loading of KY386 compound in the formulation

[0170]

[0171]

[0172] 4. Analysis of self-emulsifying particle size of different formulations

[0173] To further analyze the solubility of the oral formulation in aqueous solution, a particle size analyzer was used to analyze the particle size of the oral formulation after self-emulsification in aqueous solution. The self-emulsification particle size analysis results of the oral formulations with different formulations are shown in Table 10.

[0174] Table 9. Oral dosage form information and results

[0175] PEG400 addition amount 150μL KY386 25mg Propylene glycol monocaprylate 50 μL Polyoxyethylene (40) hydrogenated castor oil (mL) 300 μL Sample size ~0.5mL

[0176] To analyze the stability of the oral formulation in aqueous solution, we first mixed the oral formulation with purified water at a ratio of 1:1000, stirred and mixed them evenly to form a self-emulsifying system, and allowed to stand for 12 hours. The homogeneity and phase separation of the solution were observed at 0 and 12 hours. The results showed that the solution was homogeneous and did not produce phase separation or precipitation, indicating that the formulation had a relatively stable dispersion in the self-emulsifying aqueous solution ( Figure 2 ). Subsequently, the oral preparation was mixed with a pH 3.0 room temperature aqueous solution at a ratio of 1:1000 to form a self-emulsifying system. The average particle size was found to be about 20 nm, less than 100 nm, and the Zeta potential was substantially greater than 10, indicating that the self-emulsifying system formed by the oral preparation in the pH 3.0 aqueous solution was relatively stable. The oral preparation maintained stability in the acidic aqueous solution for at least 12 hours. The solution at 0 and 12 hours after the oral preparation was dissolved was as follows: Figure 2 The specific particle size distribution of the oral preparation in acidic aqueous solution is shown as follows. Figure 3 shown.

[0177] In contrast, KY386 was dissolved in a PEG400:Tween 80 = 2:1 (volume ratio) or a PEG400:Tween 80:Cremophor ELP = 2:2:1 (volume ratio) system to obtain a 50 mg / mL formulation. Analysis of the aqueous solution stability of the oral formulation using the above dilution method revealed instability and precipitation, as shown in Table 10.

[0178] Table 10. Comparison of the stability of various formulations in aqueous solution

[0179]

[0180]

[0181] NA: Not applicable

[0182] As can be seen from Table 10, the preparation using PEG400:propylene glycol monocaprylate:polyoxyethylene (40) hydrogenated castor oil = 3:1:6 maintains good aqueous solution solubility and particle size stability in acidic aqueous solution.

[0183] 5. Dissolution Analysis of KY386 Compound in Oral Formulations

[0184] The oral preparation was prepared according to the method in Table 9, and then filled into gelatin capsules. The dissolution data at different time points were obtained by dissolution test analysis, as shown in the following table.

[0185] Table 11. Summary of dissolution data of oral dosage forms at different time points

[0186]

[0187] Dissolution curve Figure 4 The dissolution results showed that KY386 could be rapidly dissolved in the dissolution solution and maintained a stable dissolution trend within 60 minutes.

[0188] 6. Stability Analysis of Oral Preparations

[0189] The oral preparation was prepared according to the method in Table 9 and then filled into liquid-filled gelatin capsules. To investigate factors affecting the stability of the oral preparation, the oral preparation was stored at high temperature (50°C), accelerated temperature (40°C), and light conditions for 30 days, and the content and impurity changes of the oral preparation were tested over 30 days. The specific preparation method of the oral preparation is shown in Table 9, and the stability test results under different storage conditions are shown in Table 12. The KY386 content in the self-emulsifying liquid was relatively stable over 30 days, while the total impurity content increased to a certain extent under all conditions. The higher the temperature, the greater the impact on the impurity content.

[0190] Table 12. Stability analysis of oral preparations under different influencing factors for 30 days

[0191]

[0192]

[0193] ND: Not Detected

[0194] Summarizing all the experimental data above, it is shown that the oral formulation, which contains KY386, propylene glycol monocaprylate, polyethylene glycol, and polyoxyethylene (40) hydrogenated castor oil, can ensure that KY386, in addition to forming a self-emulsifying body, has a particle size of less than 100 nm, and shows a good compound dissolution curve, particle size uniformity, aqueous solubility, and stability in acidic aqueous solution under an acidic environment. The oral formulation also has good stability. Under the influencing factors, KY386 does not undergo significant degradation and no significant impurities are generated, and has good stability, aqueous solubility, and dissolution properties in simulated gastric fluid.

Claims

1. An oral preparation of a polycyclic compound, characterized in that: The oral preparation contains the polycyclic compound shown in formula I, polyethylene glycol, propylene glycol monocaprylate and polyoxyethylene (40) hydrogenated castor oil.

2. The oral preparation according to claim 1, characterized in that The concentration of the polycyclic compound represented by Formula I in the oral preparation is about 0.1-80 mg / mL.

3. The oral preparation according to claim 1, characterized in that Based on the total volume of the oral preparation, the volume ratio of the polyethylene glycol is 9-81%, the volume ratio of the propylene glycol monocaprylate is about 10-30%, and the volume ratio of the polyoxyethylene (40) hydrogenated castor oil is about 9-81%.

4. The oral preparation according to claim 3, characterized in that The polyethylene glycol is PEG 300, PEG 400 or a mixture thereof, or PEG 4000, PEG 6000 or a mixture thereof.

5. The oral preparation according to claim 1, characterized in that The oral formulation also contains an antioxidant.

6. The oral preparation according to claim 5, characterized in that The mass ratio of the antioxidant is about 0.02-0.1% based on the total mass of the oral preparation.

7. A method for preparing the oral preparation of a polycyclic compound according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1. Heat polyethylene glycol to 60 ± 5°C, maintain constant temperature, and maintain nitrogen bubbling (0.03 MPa) for at least 20 minutes; S2. Add polyoxyethylene (40) hydrogenated castor oil under stirring, continue heating to 60 ± 5 ° C, and stir until fully dissolved; S3. Add the polycyclic compound, maintain nitrogen bubbling (0.03Mpa) and stir thoroughly for 60-120 minutes until the compound is dissolved, and stop heating; S4. Add propylene glycol monocaprylate, continue stirring and nitrogen aeration for 30 minutes until the mixture is uniform, and obtain the oral preparation of the polycyclic compound.

8. The method according to claim 7, characterized in that The polyethylene glycol is PEG300, PEG400 or a mixture of the two, or PEG4000, PEG6000 or a mixture of the two.

9. The method according to claim 7, characterized in that Based on the total volume of the oral preparation, the volume ratio of the polyethylene glycol is about 9-81%, the volume ratio of the propylene glycol monocaprylate is about 10-30%, and the volume ratio of the polyoxyethylene (40) hydrogenated castor oil is about 9-81%.

10. The method according to claim 7, characterized in that The method further comprises adding an antioxidant in step S3, wherein the mass ratio of the antioxidant is about 0.02-0.1% based on the total mass of the oral preparation.

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