WO2023241652A1 - Pharmaceutical composition, method for preparing same, and use thereof - Google Patents

Abstract

The present invention provides a pharmaceutical composition, a method for preparing same, and use thereof. The pharmaceutical composition comprises a compound represented by formula I. The pharmaceutical composition of the present invention has high drug loading capacity, good uniformity, and good stability, and can be used for preparing a medicament for preventing and/or treating glaucoma or ocular hypertension.

Description

Pharmaceutical composition and preparation method and application thereof Technical field

The invention belongs to the field of medical technology, and specifically relates to a pharmaceutical composition and its preparation method and application.

Background technique

Rho associated protein kinase (ROCK), a serine/threonine protein kinase, is a downstream target effector molecule of RHO and is widely expressed in the human body. RHO-related protein kinase (ROCK) participates in the regulation of myosin light chain (MLC) and is suitable for the treatment of vasodilation. ROCK can also act on trabecular outflow tract cells, relax trabecular cells, and reduce aqueous humor outflow resistance. The latest research shows that ROCK inhibitors can also promote the damage repair of corneal endothelial cells and prevent fibrosis, which has huge application prospects.

Isoquinoline sulfonamide compounds are an important type of ROCK inhibitors. Fasudil and K-115, which are currently on the market, are both isoquinoline sulfonamide compounds (WO2006057397A1). Among them, fasudil is a new drug with a wide range of pharmacological effects. It is an inhibitor of RHO kinase. It dilates blood vessels by increasing the activity of myosin light chain phosphatase, reduces the tension of endothelial cells, improves brain tissue microcirculation, and does not produce It can also aggravate brain hemorrhage, antagonize inflammatory factors, protect nerves from apoptosis, and promote nerve regeneration. The approved and potential applications of K-115 are very wide, including glaucoma, intraocular hypertension, complications of diabetic retinal damage, age-related macular degeneration, corneal damage, recovery after cataract and glaucoma surgery, etc., and may be further expanded to the system. Sex drugs.

WO2020253882A1 discloses an isoquinolinone derivative as a ROCK protein kinase inhibitor and its application in preparing drugs for ROCK protein kinase inhibitor-related glaucoma or ocular hypertension diseases, and specifically discloses compound 63 in the specification. ,

This compound shows a better intraocular pressure-lowering effect than K-115, but due to its poor water solubility, it may have many adverse effects on drugability, such as: 1) the exposure of the compound is reduced, which affects the efficacy of the drug; 2 ) Affects the metabolism of the drug in the body; 3) The dosage needs to be increased to achieve the drug effect, which will cause the drug to accumulate or crystallize in the body, increasing the risk of toxic side effects; 4) It is not easy to make oral or intravenous or other liquid preparations , resulting in an increase in R&D investment in the later period; especially in the field of ophthalmic preparations, the range of dosage forms to choose from is narrow (mainly liquid preparations, gels, eye ointments, etc.). Among them, eye drops are the most ideal dosage form, but due to This compound has poor water solubility, and often only a suspension can be obtained, or the uniformity and stability indicators of the developed liquid preparation are difficult to meet the requirements. It has the disadvantages of large eye irritation and poor patient compliance, and is not suitable for use as a medicine for patients. use.

Therefore, in order to overcome the above shortcomings, it is very necessary to develop ophthalmic preparations, especially liquid preparations, that are suitable for pharmaceuticals and have simple components to improve the solubility of the compounds, preparation stability and patient compliance.

Contents of the invention

In view of the shortcomings of the existing technology, the object of the present invention is to provide a pharmaceutical composition and its preparation method and application. The pharmaceutical composition includes a sulfate form of an isoquinolinone derivative, which has higher drug loading capacity, better uniformity and stability.

In a first aspect, the invention provides a pharmaceutical composition comprising a compound of formula I:

In some embodiments of the invention, the pharmaceutical composition includes a compound of formula I and a pharmaceutically acceptable Acceptable excipients, carriers and/or diluents.

In some embodiments of the invention, the pharmaceutical composition includes a compound of Formula I and a solubilizing agent. In some embodiments of the invention, the pharmaceutical composition includes a compound of Formula I and an osmotic pressure regulator. In some embodiments of the invention, the pharmaceutical composition includes a compound of Formula I and a bacteriostatic agent.

In some embodiments of the present invention, the pharmaceutical composition includes a compound of Formula I, a solubilizer, an osmolality regulator, and a bacteriostatic agent.

In the pharmaceutical composition, the mass ratio of the compound of formula I to the osmotic pressure regulator can be 1:(4.5-600), such as 1:(10-500), 1:(20-400), 1:(30- 350) etc.

In the pharmaceutical composition, the mass ratio of the compound of formula I to the solubilizer can be 1:(2-70), preferably 1:(2-50), such as 1:(2.5-40), 1:(5- 40), 1:(10-40), etc.

In some embodiments of the present invention, the solubilizing agent is selected from Tween 80, polyoxyethylene 40 hydrogenated castor oil, polyethylene glycol 400, poloxamer 188, polyoxyethylene hydrogenated stearate, polyoxyethylene One or more of ethylene castor oil and its derivatives, such as one, two, three or four.

In some embodiments of the invention, the solubilizer is polyoxyethylene hydrogenated castor oil.

In some embodiments of the present invention, the polyoxyethylene hydrogenated castor oil is selected from one or more of polyoxyethylene 40 hydrogenated castor oil, polyoxyethylene 54 hydrogenated castor oil, and polyoxyethylene 60 hydrogenated castor oil.

In some embodiments of the present invention, the solubilizing agent is Tween 80 and/or polyoxyethylene 40 hydrogenated castor oil.

In some embodiments of the present invention, the solubilizer is Tween 80 and polyoxyethylene 40 hydrogenated castor oil. In some embodiments, in the composition, the mass ratio of Tween 80 and polyoxyethylene 40 hydrogenated castor oil is 1:(0.1-30), such as 1:(0.5-20), 1:(1- 10), 1:(2-10), 1:(2-8), etc.

In some embodiments of the present invention, the osmotic pressure regulator is selected from one or more of sodium chloride, potassium chloride, mannitol, glycerin, propylene glycol, dextrose, boric acid and borax, such as a , two, three or four. Preferably, the osmotic pressure regulator is sodium chloride and/or mannitol. More preferably, the osmotic pressure regulator is sodium chloride and mannitol.

In some embodiments of the present invention, the bacteriostatic agent is selected from benzalkonium chloride, thimerosal, dumycin Any combination of one or more of phenylene, xanthanol, chlorobutanol, parabens, and sorbic acid is preferably benzalkonium chloride.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, and a compound selected from the group consisting of Tween 80, polyoxyethylene 40 hydrogenated castor oil, sodium chloride, mannitol and benzalkonium chloride. one or more.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, and one or more of polyoxyethylene hydrogenated castor oil, sodium chloride, mannitol, and benzalkonium chloride.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, and polyoxyethylene hydrogenated castor oil.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, polyoxyethylene hydrogenated castor oil, sodium chloride, mannitol, and benzalkonium chloride.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, polyoxyethylene 40 hydrogenated castor oil, sodium chloride, mannitol, and benzalkonium chloride.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, Tween 80, and polyoxyethylene 40 hydrogenated castor oil.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, and sodium chloride.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, sodium chloride, and benzalkonium chloride.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, sodium chloride, and mannitol.

In some embodiments of the invention, the pharmaceutical composition includes the following components: a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, sodium chloride, mannitol, and benzalkonium chloride.

In some embodiments of the present invention, in the pharmaceutical composition, the weight parts of the compound of formula I, solubilizer, osmotic pressure regulator and bacteriostatic agent are respectively:

In some embodiments of the present invention, in the pharmaceutical composition, the weight portion of the solubilizer can be any value in the range of 2-70 parts, for example, it can be 2 parts, 3 parts, 4 parts, or 5 parts. , 6 servings, 8 servings, 10 servings, 15 servings, 20 servings, 25 servings, 30 servings, 35 servings, 40 servings, 45 servings, 50 servings, 55 servings, 60 servings, 65 servings or 70 servings.

In some embodiments of the present invention, in the pharmaceutical composition, the weight parts of the osmotic pressure regulator can be any value in the range of 13-600 parts, for example, it can be 13 parts, 14 parts, 15 parts, 16 copies, 18 copies, 20 copies, 25 copies, 30 copies, 40 copies, 50 copies, 60 copies, 70 copies, 80 copies, 90 copies, 100 copies, 120 copies, 150 copies, 180 copies, 200 copies, 250 copies, 300 copies, 350 copies, 400 copies, 450 copies, 500 copies, 550 copies or 600 copies.

In some embodiments of the present invention, in the pharmaceutical composition, the weight part of the bacteriostatic agent can be any value in the range of 0.05-3.2 parts, for example, it can be 0.05, 0.1 part, 0.2 part, 0.3 part, 0.4 parts, 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, 0.9 parts, 1 part, 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts , 2.1 parts, 2.2 parts, 2.3 parts, 2.4 parts, 2.5 parts, 2.6 parts, 2.7 parts, 2.8 parts, 2.9 parts, 3 parts, 3.1 parts or 3.2 parts.

In some embodiments of the present invention, the pharmaceutical composition includes the following components by weight:

1 part of compound of formula I;

Tween 80 0.3-3.5 parts (such as 0.3 parts, 0.4 parts, 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, 0.9 parts, 1 part, 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts , 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, 2.1 parts, 2.2 parts, 2.3 parts, 2.4 parts, 2.5 parts, 2.6 parts, 2.7 parts, 2.8 parts, 2.9 parts, 3 parts, 3.2 parts, etc.); and

Polyoxyethylene hydrogenated castor oil such as polyoxyethylene 40 hydrogenated castor oil 2-35 parts (such as 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 servings, 13 servings, 14 servings, 15 servings, 16 servings, 17 servings, 18 servings, 19 servings, 20 servings, 21 servings, 22 servings, 23 servings, 24 servings, 25 servings, 26 servings, 27 servings, 28 servings , 29 copies, 30 copies, 32 copies, etc.).

In some embodiments of the present invention, the pharmaceutical composition includes the following components by weight:

1 part of compound of formula I;

Tween 80 0.3-3.5 parts (such as 0.4 parts, 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, 0.9 parts, 1 part, 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, 2.1 parts, 2.2 parts, 2.3 parts , 2.4 parts, 2.5 parts, 2.6 parts, 2.7 parts, 2.8 parts, 2.9 parts, 3 parts, 3.2 parts, etc.);

Polyoxyethylene hydrogenated castor oil such as polyoxyethylene 40 hydrogenated castor oil 2-35 parts (such as 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 servings, 13 servings, 14 servings, 15 servings, 16 servings, 17 servings, 18 servings, 19 servings, 20 servings, 21 servings, 22 servings, 23 servings, 24 servings, 25 servings, 26 servings, 27 servings, 28 servings , 29 copies, 30 copies, 32 copies, etc.);

Sodium chloride 9-110 parts (such as 9 parts, 10 parts, 13 parts, 15 parts, 18 parts, 20 parts, 25 parts, 30 parts, 35 parts, 40 parts, 50 parts, 60 parts, 70 parts, 80 parts , 90 copies, 100 copies or 107 copies, etc.); and

Mannitol 4-200 parts (such as 4 parts, 5 parts, 8 parts, 10 parts, 12 parts, 15 parts, 18 parts, 20 parts, 25 parts, 30 parts, 40 parts, 50 parts, 60 parts, 80 parts, 100 copies, 120 copies, 150 copies, 170 copies, 190 copies, 195 copies, etc.).

In some embodiments of the present invention, the pharmaceutical composition includes the following components by weight:

In some embodiments of the present invention, the dosage form of the pharmaceutical composition is a solution, suspension, emulsion, gel, micelle or any pharmaceutically applicable ophthalmic topical dosage form.

In some embodiments of the present invention, the content of the compound of formula I in the pharmaceutical composition is 0.01-2 mg/mL, specifically, it can be any value in the range of 0.01-2 mg/mL, for example, it can be 0.01 mg/mL. , 0.02mg/mL, 0.05mg/mL, 0.08mg/mL, 0.1mg/mL, 0.2mg/mL, 0.3mg/mL, 0.5mg/mL, 0.8mg/mL, 1mg/mL, 1.2mg/mL, 1.5 mg/mL, 1.8 mg/mL or 2 mg/mL, etc.; preferably any value within the range of 0.05-1 mg/mL.

In some embodiments of the present invention, the crystal form of the compound of formula I is crystal form A or crystal form B.

In some embodiments of the present invention, the crystal form of the compound of formula I is crystal form B.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 6.33±0.20°, 10.62±0.20°, and 13.11±0.20°.

In some aspects of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 3.30±0.20°, 6.33±0.20°, 6.55°±0.20°, 10.62±0.20°, 12.57 ±0.20°, 13.11±0.20°.

In some aspects of the present invention, the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2θ angles: 3.30±0.20°, 6.33±0.20°, 10.62±0.20°, 12.57±0.20°, 13.11± 0.20°, 17.85±0.20°, 18.51±0.20°, 20.99±0.20°.

In some aspects of the invention, the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2θ angles: 3.30±0.20°, 6.33±0.20°, 6.55±0.20°, 10.62±0.20°, 12.57± 0.20°, 13.11±0.20°, 14.20±0.20°, 16.37±0.20°, 17.85±0.20°, 18.51±0.20°, 19.56±0.20°, 20.99±0.20°, 25.53±0.20°, 26.35±0.20°.

In some aspects of the present invention, the X-ray powder diffraction pattern of the above-mentioned B crystal form has characteristic diffraction peaks at the following 2θ angles: 9.69±0.20°, 12.12±0.20°, 16.48±0.20°, 16.95±0.20°, 17.94± 0.20°, 19.23±0.20°, 20.37±0.20°, 21.87±0.20°.

The invention provides the B crystal form of the compound of formula I, the X-ray powder diffraction pattern of which has characteristic diffraction peaks at the following 2θ angles: 16.48±0.20°, 16.95±0.20°, and/or 21.87±0.20°, and/or 12.12 ±0.20°, and/or 17.94±0.20°, and/or 9.69±0.20°, and/or 20.37±0.20°, and/or 21.87±0.20°, and/or 4.80±0.20°, and/or 14.61±0.20 °, and/or 19.23±0.20°, and/or 27.53±0.20°, and/or 28.72±0.20°, and/or 33.61±0.20°.

In some aspects of the invention, the X-ray powder diffraction pattern of the above-mentioned B crystal form has characteristic diffraction peaks at the following 2θ angles: 4.80±0.20°, 9.69±0.20°, 12.12±0.20°, 14.61±0.20°, 16.48± 0.20°, 16.95±0.20°, 17.94±0.20°, 19.23±0.20°, 20.37±0.20°, 21.87±0.20°, 27.53±0.20°, 28.72±0.20°, 33.61±0.20°.

In some aspects of the present invention, the X-ray powder diffraction pattern of the above-mentioned B crystal form has characteristic diffraction peaks at the following 2θ angles: 4.80±0.20°, 9.69±0.20°, 12.12±0.20°, 16.48±0.20°, 16.95± 0.20°, 17.94±0.20°, 19.23±0.20°, 20.37±0.20°, 21.87±0.20°.

In some embodiments of the present invention, the pH of the pharmaceutical composition is 4.0-6.0, specifically, it can be any value in the range of 4.0-6.0, for example, it can be 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, or 6.0.

In some preferred embodiments of the present invention, by selecting Tween 80 and/or polyoxyethylene 40 hydrogenated castor oil as solubilizers, and/or selecting sodium chloride and mannitol as osmotic pressure regulators for synergy, further improving The solubility of the compound of formula I and the stability of the liquid preparation containing the compound can help solve the shortcomings of low drug loading, poor uniformity and stability of the preparation, etc. caused by the poor water solubility of the compound of formula I.

In a second aspect, the present invention provides a method for preparing the pharmaceutical composition as described in the first aspect, comprising the following steps:

The compound of formula I is mixed with pharmaceutically acceptable excipients, carriers and/or diluents (such as solubilizers, osmotic pressure regulators and/or bacteriostatic agents) in corresponding parts by weight to obtain the pharmaceutical composition.

Preferably, the preparation method includes: mixing the compound of formula I with a solubilizer, an osmotic pressure regulator and a bacteriostatic agent in corresponding parts by weight to obtain the pharmaceutical composition.

In some embodiments of the present invention, the preparation method includes: adding the compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, optionally an osmotic pressure regulator and optionally a bacteriostatic agent in corresponding parts by weight. Mix evenly to obtain the composition.

It should be noted that "optionally" mentioned in the present invention means with or without corresponding components. When the pharmaceutical composition contains the corresponding component, the component is used in the preparation method; when the pharmaceutical composition does not contain the corresponding component, the component is not used in the preparation method.

In some embodiments of the invention, the preparation method includes the following steps:

(1) Dissolve the osmotic pressure regulator with water for injection to obtain solution I;

(2) Mix the solubilizer and the compound of formula I evenly to obtain mixture II;

(3) Mix solution I and mixture II and stir evenly to obtain solution III;

(4) Dissolve the bacteriostatic agent in water for injection to obtain solution IV;

(5) Mix the solution III and the solution IV, dilute to full volume with water for injection, and perform three-stage filtration and sterilization with a microporous filter membrane to obtain the pharmaceutical composition.

In a third aspect, the present invention provides the pharmaceutical composition as described in the first aspect or the pharmaceutical composition prepared by the preparation method as described in the second aspect in the preparation of medicines for preventing and/or treating glaucoma or ocular hypertension. application.

Compared with the prior art, the present invention has the following beneficial effects:

The compound of formula I of the present invention is in the form of a sulfate salt. Compared with its non-salt form, the water solubility is improved, and the obtained composition has a higher drug loading capacity, better uniformity and stability.

In the embodiment of the present invention, by selecting Tween 80 and/or polyoxyethylene 40 hydrogenated castor oil as the solubilizer, the solubility of the compound of formula I and the stability of the pharmaceutical preparation containing the compound are further improved, and the problem of formula I is solved. The poor water solubility of the compound results in low drug loading, poor uniformity and stability of the preparation, and other defects that lead to poor finished drugs. The composition thus obtained is capable of completely dissolving the compound of formula I for 12 months long term (25°C ± 2°C; RH 40% ± 5%, 5 ± 3°C) and 6 months accelerated (40°C ± 2°C; RH 25% ± 5 %) conditions, the preparation always remains colorless and clear, and the osmotic pressure, pH value, relative content of the compound of formula I, and the content of related substances are all relatively stable.

Detailed ways

The technical solution of the present invention will be further described below through specific implementations. Those skilled in the art should understand that the specific embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

X-ray powder diffraction (X-ray powder diffractometer, XRPD) method

Instrument model: PANalytical X'pert ³ X-ray powder diffractometer

Test method: About 10mg sample is used for XRPD detection.

The detailed XRPD parameters are as follows:

Ray source: Cu, Cu,

Light tube voltage: 40kV, light tube current: 40mA

Scanning range: 3-40deg

Step width angle: 0.0263deg

Step length: 46.665 seconds

Synthesis of intermediates 1-4

first step

Control the internal temperature to 25-35°C, add toluene (24L) to the reaction kettle, and add compound 1-4a (4000g), sodium carbonate (6120g), methylboronic acid (3465g), and 2-dimethylboric acid (3465g) to the reaction kettle in sequence while stirring. Cyclohexylphosphine-2', 6'-dimethoxybiphenyl (98.66g) and tridibenzylideneacetone dipalladium (88.03g), replace the reaction kettle with nitrogen three times, and raise the internal temperature of the reaction kettle to 90°C , and stir at 90 to 100°C for 13 hours. Add water (4L) to the reaction kettle to dissolve the solid, cool the reaction liquid to room temperature, filter the reaction liquid through diatomaceous earth, and wash the filter cake with methyl tert-butyl ether (4L). Combine the washing liquid and the filtrate and use concentrated hydrochloric acid. (8L) Adjust pH = 3 and separate liquids at rest. The lower aqueous phase was extracted with methyl tert-butyl ether (5L), the pH of the aqueous phase was adjusted to 9 with aqueous sodium hydroxide solution, the aqueous phase was extracted twice with ethyl acetate (12L), and the combined organic phases were concentrated to weight under vacuum. Compound 1-4b was obtained without further reduction.

MS-ESI calculated value [M+H] ⁺ 144, measured value 144.

Step 2

Control the temperature inside the reaction kettle to be lower than 30°C, and slowly add concentrated sulfuric acid (13.36L) into the 50L reaction kettle. Control the internal temperature below 50°C, and slowly add compound 1-4b (2500g) into the reaction kettle using a constant pressure dropping funnel. Control the internal temperature of the reaction kettle to -10~0°C and slowly add N-bromosuccinimide (3070.25g) in batches to the reaction kettle, and stir the reaction solution in the range of -10~8°C for 13 hours. Control the internal temperature below 50°C and slowly pour the reaction solution into ice water (8L). Control the internal temperature below 50°C, slowly add sodium hydroxide aqueous solution dropwise to the reaction solution, and adjust pH=9. Ethyl acetate (10 L) was added to the reaction solution and stirred for 10 minutes. The reaction liquid was filtered, and the filter cake was washed with ethyl acetate (12.5L). The aqueous phase was extracted with ethyl acetate (10L×3), and the combined organic phases were concentrated under reduced pressure at 45°C. Add n-heptane (10L) to the concentrated residue for pulping, and perform suction filtration to obtain a brown-red solid. The solid was dried in a vacuum drying oven (40°C) to obtain compound 1-4c.

MS-ESI calculated value [M+H] ⁺ 222,224, measured value 222,224.

third step

Add the solvent N, N-dimethylacetamide (10L) to the reaction kettle (50L), add 1-4c (1003.65g) under stirring, and control the internal temperature below 55°C to slowly add the solvent to the reaction kettle (50L) in batches. Sodium methylmercaptide (1276.97g) was added and the reaction mixture was stirred at 50°C for 30 minutes. Raise the temperature of the reaction kettle to 120°C. At this time, the internal temperature is 109°C. Stir under this condition for 12 hours. Adjust the temperature of the reaction kettle to 50°C and lower the temperature of the reaction solution to 40~50°C. Filter the reaction solution through diatomaceous earth and wash it with ethyl acetate (10L). Add water (10L) to the filtrate and wash it with ethyl acetate. (5L×2) extract the aqueous phase. Adjust the pH of the aqueous phase to 7 with concentrated hydrochloric acid, extract the aqueous phase with ethyl acetate (5L×2), combine the organic phases, wash the organic phase with water (10L×3), and concentrate the organic phase under reduced pressure until there is no change in weight to obtain the compound 1-4d.

MS-ESI calculated value [M+H] ⁺ 176, measured value 176.

the fourth step

Add dichloromethane (8L) into the cleaned 50L high and low temperature reaction kettle, and start stirring. Add compound 1-4d (1474.02g) into the reaction kettle, adjust the temperature of the oil bath to control the temperature in the reaction kettle at 0-10°C. Slowly add concentrated hydrochloric acid (4.38L) into the reaction kettle in batches, and control the internal temperature at 5 to 15°C. After the dropwise addition of concentrated hydrochloric acid is completed, lower the internal temperature of the reaction kettle to -5°C. The sodium hypochlorite aqueous solution (21.35L) was slowly added dropwise into the reaction kettle in batches, and the temperature of the entire process was controlled at 0 to 10°C. The reaction liquid was filtered, the filter cake was washed with methyl tert-butyl ether (2L×2), and the filter cake was collected. The collected filter cake was dried in an oven to obtain compound 1-4e.

MS-ESI calculated value [M+H] ⁺ 242, measured value 242.

the fifth step

Add methylene chloride (10L) into the 50L reaction kettle, start stirring, control the internal temperature to 0-10°C, weigh out 1-4e (1462.35g) and add it into the reaction kettle. Control the internal temperature from 0 to 10°C, weigh out N, N-diisopropylethylamine (934.63g) and slowly add it dropwise into the reaction solution. Control the internal temperature from 0 to 10°C and weigh out 1-4f (1230.24g). Add slowly into the reaction solution in batches. After the addition is completed, raise the temperature to 10-20°C and continue stirring for 0.5 hours. The organic phase was washed with saturated aqueous ammonium chloride solution (3L×3), and the combined saturated aqueous ammonium chloride solution was extracted with dichloromethane (3L). The organic phase was separated and concentrated under reduced pressure until there was no change in weight, and then dried in a vacuum drying oven to obtain compound 1-4g.

MS-ESI calculated value [M+H] ⁺ 392, measured value 392.

Step 6

Add 3.1L of anhydrous dichloromethane into the cleaned 5L three-necked bottle and start stirring. Add the weighed 1-4g (451.34g) into the reaction bottle. After the solid is dissolved, put the reaction bottle into an ice water bath to reduce the internal temperature of the reaction system to 0°C. Slowly add the weighed m-chloroperoxybenzoic acid (429.81g) into the 5L reaction bottle in batches, keeping the temperature at 0-10°C throughout the process. After the addition of m-chloroperoxybenzoic acid is completed, remove the ice-water bath from the reaction bottle and replace it with an oil bath to stabilize the internal temperature at 20-25°C and stir for 12 hours. Put the reaction bottle into an ice water bath and stir for 1 hour, filter, rinse the filter cake with methylene chloride (400mL turns blue, add saturated aqueous sodium bicarbonate solution dropwise to the system until pH=7. Steady layering, retain the organic phase, extract the aqueous phase again with 3L of methylene chloride, and combine the organic phases. The organic phase was washed with 1% sodium bicarbonate aqueous solution (4L×10), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The solid obtained by concentration was dried in an oven to obtain compound 1-4h.

MS-ESI calculated value [M+H] ⁺ 408, measured value 408.

Step 7

Add 1-4h (301.42g) to a 5L three-necked flask, add tetrahydrofuran (1.5L), stir evenly, add triethylamine (206mL) dropwise at an internal temperature of 10 to 20°C, and stir evenly. Trifluoroacetic anhydride (206 mL) was added dropwise at an internal temperature of 20 to 30°C and reacted at 20 to 30°C for 0.5 hours. Add water (1.5L) to the reaction solution and stir evenly until the reaction solution becomes turbid. Add solid sodium hydroxide (600g) at an internal temperature of 40 to 50°C and stir for 0.5 hours. A large amount of solid precipitates. Add 1.5L of methyl tert-butyl ether and stir for 0.5 hours. The reaction solution was separated, and the upper organic phase was filtered to obtain a solid. The solid was beaten with methyl tert-butyl ether/ethyl acetate (4.4L, 10:1), and the beaten liquid was filtered to obtain a white solid. The white solid was dried in an oven to obtain crude product 1-4 (371.52g). Add tetrahydrofuran (7L) to the 50L reaction kettle, control the internal temperature to 20-25°C, add crude compound 1-4 (1713.19g, combine other batches), and stir for 0.5 hours at an internal temperature of 20-25°C. Add 8.6L of water and stir at 20-25°C for 0.5 hours. Add solid sodium hydroxide (800g) and stir at an internal temperature of 20-25°C for 0.5 hours. Add methyl tert-butyl ether (8.6L) and stir for 0.5 hours at an internal temperature of 20 to 25°C. Separate the liquids, filter the upper organic phase, retain the solid obtained during filtration, and dry the solid in an oven to constant weight to obtain compound 1-4.

MS-ESI calculated value [M+H] ⁺ 408, measured value 408.

Preparation of crystal form A of compound of formula I

first step

Turn on stirring and nitrogen protection, and add methylene chloride (8.6L), water (8.6L), and starting material compound 1-1 (575.34g) into the reaction kettle in sequence. Control the internal temperature of the reaction kettle to 10-15°C, and add sodium bicarbonate (1289.36g), tetrabutylammonium bisulfate (133.87g) and starting material compound 1-2 (760.25g) to the reaction kettle in sequence. The temperature is 10-15°C and stirred for 2 hours. The reaction solution was allowed to stand and separated into layers, and the organic phase was concentrated under reduced pressure until no fraction flowed out. Transfer the concentrated solution to a three-necked flask and control the internal temperature to 60-70°C. After stirring for 12 hours, cool to room temperature naturally. The crude product is quickly filtered through silica gel and rinsed with dichloromethane until no product remains. The eluate was concentrated under reduced pressure to about 1.2L, washed with 1% sodium bicarbonate aqueous solution (3L), the organic phase was dried over anhydrous sodium sulfate (500g), filtered, and then concentrated under reduced pressure until no fraction was found. Until flowing out, the intermediate 1-3 was obtained and used directly in the next step without purification.

MS-ESI calculated value [M+H] ⁺ 199, measured value 199.

¹ H NMR (400MHz, CD ₃ Cl) δ7.81-7.79 (d, J = 8Hz, 1H), 7.01-6.95 (m, 2H), 5.84 (s, 2H), 2.52 (s, 3H), 2.27 ( s,3H) ppm.

Step 2

Add 2-methyltetrahydrofuran (9702mL) to the reaction kettle, start stirring and control the internal temperature to 10-20°C, then add intermediate 1-4 (1078.26g), cesium carbonate (1017.63g) and intermediate 1-3 in sequence (631.71g). The temperature inside the reaction kettle was raised to 57-63°C and the reaction was stopped after stirring for 2 hours and 40 minutes. Lower the internal temperature of the reaction kettle to 15-25°C and add water (10.78L) to the reaction solution. Stir and let stand for separation. The resulting aqueous phase is extracted twice with 2-methyltetrahydrofuran (5390mL×2). Combine the The organic phase was concentrated under reduced pressure to about 2.2L. Add acetone (1078mL) and n-heptane (2156mL) to the concentrated solution And continue to concentrate until no fraction flows out, and then a loose solid can be obtained. Control the internal temperature to 10-30°C, beat the solid with a mixed solvent (11.0495L, acetone:n-heptane=1:40) and stir for three times, filter and rinse with n-heptane (1078mL). The filter cake is dried under vacuum to constant weight to obtain intermediate 1-5.

MS-ESI calculated value [M+H] ⁺ 570, measured value 570.

¹ H NMR (400MHz, CD ₃ Cl) δ8.86-8.78(m,1H),8.13-8.04(m,1H),7.93-7.86(m,1H),7.58-7.49(m,1H),7.39- 7.32(m,1H),7.09-6.99(m,2H),6.14(s,2H),4.86-4.76(m,1H),4.42-4.30(m,1H),3.73-3.61(m,2H), 3.57-3.47(m,1H),3.46-3.37(m,1H),2.76-2.67(m,3H),2.59(s,3H),2.39-2.29(m,4H),2.09-1.97(m,1H ),1.46(s,9H)ppm.

third step

Control the internal temperature to 20-30°C, add anhydrous tetrahydrofuran (4.24L) and concentrated sulfuric acid (712.43g) to a 5L three-necked flask in sequence, stir evenly and set aside. Control the internal temperature to 20-30°C and add intermediate 1-5 (1062.15g) and tetrahydrofuran (4.24L) to the reaction kettle in sequence. Stir evenly and add the pre-configured sulfuric acid tetrahydrofuran solution dropwise into the system. The temperature was controlled at 35-45°C and stirred for 4 hours. A large amount of white precipitate gradually precipitated in the system. Slowly add methyl tert-butyl ether (8.48L) into the reaction kettle, control the internal temperature at 20-30°C and stir for 0.5 hours. The filtered solid is dissolved in a mixed solvent (methyl tert-butyl ether/methyl tert-butyl ether/ Tetrahydrofuran = 10.6L/10.6L) and methyl tert-butyl ether (21.2L) were sequentially beaten for 0.5 hours, and then beaten with water five times (21.2L × 5) until the pH of the filtrate was about 7. Control the internal temperature to 20-30°C, beat the solid in methyl tert-butyl ether (15.9L) and mixed solvent (methyl tert-butyl ether/tetrahydrofuran = 10.6L/5.3L) in sequence, filter, Rinse and dry under vacuum below 45°C to obtain the crystalline form A of compound I. After ion chromatography detection, the sulfate content is 9.17%, from which it can be inferred that there is 0.5 sulfate in the compound of formula (II).

MS-ESI calculated value [M+H] ⁺ 470, measured value 470. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.64-8.58 (m, 1H), 8.26 (d, J = 8Hz, 1H), 7.74-7.62 (m, 3H), 7.16-7.11 (m, 1H) ,7.11-7.05(m,1H),6.08(s,2H),3.94-3.89(m,1H),3.70-3.55(m,2H),3.55-3.44(m,1H),3.43-3.27(m, 1H),2.59(s,3H),2.48-2.45(m,3H),2.35-2.29(m,1H),2.29-2.24(m,3H),2.16-1.90(m,1H)ppm.

The X-ray powder diffraction pattern analysis data of the crystalline form A of the compound of formula I are shown in Table 1.

Table 1 X-ray powder diffraction pattern analysis data of compound A of formula I

Preparation of crystal form B of compound of formula I

Weigh an appropriate amount of compound A crystal form of formula I and add it to the sample bottle, add a certain volume of the solvents in Table 2 to prepare suspensions or solutions of different single solvents. After the suspension was continuously stirred at 50°C for 3 hours, the sample was filtered and the filtered solid was placed in a vacuum drying oven and vacuum dried at 45°C to remove the residual solvent to obtain the crystal form B of the compound of formula I. The preparation conditions of the crystal form B of the compound of formula I are shown in Table 2 below. The X-ray powder diffraction pattern analysis data of the crystal form B of the compound of formula I are shown in Table 3.

Table 2 Preparation of crystal form B of compound I

Table 3 X-ray powder diffraction pattern analysis data of compound B crystal form of formula I

Study on Hygroscopicity of Crystal Form B of Compound I

Experimental Materials:

SMS DVS intrinsic dynamic gas adsorption meter.

experimental method:

Take 10-15 mg of the crystal form B of compound I and place it in the DVS sample tray for testing.

Experimental results:

The hygroscopic weight gain of the crystal form B of compound I at 25° C. and 80% RH is 0.736%, and it is slightly hygroscopic.

Study on solid stability of crystal form B of compound I

The crystal form B of the compound of formula I was placed for 6 months under the conditions of 40°C/75%RH (relative humidity) (double-layer LDPE bag was sealed and then aluminum foil bag was heat-sealed). Double-layer LDPE bags are sealed and then heat-sealed with aluminum foil bags) and stored for 12 months. The crystal form was tested separately at each sampling point to determine the crystal form stability of the sample. The experimental results of solid stability research on the crystalline form B of compound I are shown in Table 4.

Table 4 Solid stability research experimental results of compound B crystal form of formula I

Conclusion: Crystal form B of the compound of formula I has good stability under accelerated and long-term tests.

In the examples of the present invention, a high-performance liquid phase detector (Agilent 1260PDA/UV detector) was used to detect the content of the compound of formula I. The chromatographic conditions are as follows in Table 5.

table 5

In the following examples, unless otherwise specified, the compounds of formula I are all tested using the crystal form B of the compound of formula I.

Example 1 The solubility test of the crystalline form B of compound I is shown in Table 6.

Table 6

It can be seen from the above tests that although the water solubility of the compound of formula I is improved compared with the non-salt form, other excipients need to be added to meet the requirements of eye drop preparations.

Example 2

In this example, the compound of Formula I and the solubilizing agent were dissolved in water and prepared into different formulations to examine the effects of different solubilizing agents on the solubility of the compound of Formula I. The specific formulation ingredients and experimental results are shown in Table 7 below.

Table 7

It can be seen from the experimental results in Table 7 that in the presence of a certain content of Tween 80 and/or polyoxyethylene 40 hydrogenated castor oil (prescriptions 4, 5, and 7), the compound of formula I can not only be completely dissolved, but also can be dissolved for a long time. There will be no precipitation after storage, and the resulting prescription has good stability.

Example 3

In this example, Tween 80 and polyoxyethylene 40 hydrogenated castor oil are used as solubilizers, and an osmotic pressure regulator is further added to prepare different prescriptions. The compatibility of the osmotic pressure regulator and the solubilizer is investigated. The specific prescription ingredients and experiments are The results are shown in Table 8 below:

Table 8

It can be seen from the experimental results in Table 8 that in the presence of a certain osmotic pressure regulator and changing the proportion of solubilizing agent, the solubility and formulation stability of the compound of formula I can be greatly improved.

Example 4

In this example, Tween 80 and polyoxyethylene 40 hydrogenated castor oil were used as solubilizers, sodium chloride was used as the osmotic pressure regulator, and a bacteriostatic agent was further added to prepare different prescriptions to examine the compatibility between raw materials and auxiliary materials. , the specific prescription ingredients and experimental results are shown in Table 9 below:

Table 9

Among them, the content detection result refers to the percentage of the detected content of the compound of formula I in the prescription to the calibrated content.

It can be seen from the experimental results in Table 9 that the compatibility of raw materials and excipients is very good after adding osmotic pressure regulators (such as sodium chloride) and/or bacteriostatic agents.

Example 5

This embodiment provides several preparations including compounds of formula I. The specific prescription ingredients are shown in Table 10 below:

Table 10

The preparation method of the above prescription preparation is as follows:

(1) Dissolve sodium chloride with about 10% water for injection, and stir to dissolve it completely to obtain solution I;

(2) Dissolve mannitol with about 10% water for injection and stir to completely dissolve to obtain solution II;

(3) Weigh the prescribed amount of the compound of formula I, add the prescribed amount of Tween 80 and polyoxyethylene 40 hydrogenated castor oil, stir to fully mix, then add solution I and solution II respectively while stirring, and stir thoroughly with a high-speed mixer Homogeneously, solution III is obtained;

(4) Weigh the prescribed amount of benzalkonium chloride, add about 10% water for injection, stir and dissolve to obtain a solution IV;

(5) Mix solution III and solution IV, dilute to full volume with cold sterile water for injection, and use 0.22 μm and 0.45 μm microporous membranes for three-stage filtration and sterilization;

(6) Take the intermediate product and repack it after passing the test. The test results are shown in Table 11 below.

Table 11

Note: Compliance with regulations refers to compliance with the requirements for visible foreign matter in ophthalmic preparations of the Chinese Pharmacopoeia.

Among them, the relative content of the compound of formula I refers to the percentage of the detected content of the compound of formula I in the prescription to the calibrated content.

It can be seen from the test results in Table 11 that the properties, pH value, osmotic pressure, content of related substances, benzalkonium chloride and compound of formula I of prescriptions 19 to 23 are relatively stable, and the preparation process is simple and suitable for industrial production.

Long-term Stability Test of Formula 20 in Example 5

Seal 5 mL of prescription 20 in a low-density polyethylene medicinal eye drop bottle, and examine the prescription at 0, 3, 6, 9, and 12 months at a temperature of 25°C ± 2°C and a relative humidity of 40% ± 5%. Check whether the properties, pH value, osmotic pressure, visible foreign matter, related substances and other indicators meet the requirements. The test results are as follows in Table 12.

Table 12

"/" means not detected

It can be seen from the 12-month long-term stability test results in Table 12 that prescription 20 is stable at 25℃±2℃, Under the condition of relative humidity of 40% ± 5%, the properties remain as a colorless clear liquid, and there are no obvious changes in osmotic pressure, pH value, relative content of the compound of formula I, and content of related substances, showing good stability.

Example 6

This embodiment provides several preparations including the compound of formula I. The auxiliary materials in the preparation prescription are polyoxyethylene 40 hydrogenated castor oil, mannitol, sodium chloride, benzalkonium chloride and water for injection.

The specific prescription ingredients are shown in Table 13 below.

Table 13

The preparation method of the above prescription preparation is as follows:

(1) Dissolve sodium chloride with about 10% water for injection, and stir to dissolve it completely to obtain solution I;

(2) Dissolve mannitol with about 10% water for injection and stir to completely dissolve to obtain solution II;

(3) Weigh the prescription amount of the compound of formula I, add the prescription amount of polyoxyethylene 40 hydrogenated castor oil, stir to fully mix, then add solution I and solution II respectively while stirring, and stir thoroughly with a high-speed mixer to obtain a solution III;

(4) Weigh the prescribed amount of benzalkonium chloride, add about 10% water for injection, stir and dissolve to obtain a solution IV;

(5) Mix solution III and solution IV, dilute to full volume with cold sterile water for injection, and use 0.22 μm and 0.45 μm microporous membranes for three-stage filtration and sterilization;

(6) Take the intermediate product and repack it after passing the test.

Long-term stability testing of formulation 26

Seal 5 mL of prescription 26 in a low-density polyethylene medicinal eye drop bottle, and examine the prescription at 0, 1, 2, 3, and 6 months at a temperature of 25°C ± 2°C and a relative humidity of 40% ± 5%. properties, pH value, osmotic pressure, visible foreign matter, related substances and other indicators meet the requirements. The test results are as follows in Table 14.

Table 14

Although the present invention has been described in detail above with general descriptions, specific embodiments and tests, it is obvious to those skilled in the art that some modifications or improvements can be made on the basis of the present invention. . Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims (10)

1. A pharmaceutical composition comprising a compound of formula I,
   
2. The pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition further includes: one or more of a solubilizer, an osmotic pressure regulator and a bacteriostatic agent;

   Preferably, the pharmaceutical composition further includes a solubilizer and an osmotic pressure regulator.
3. The pharmaceutical composition according to claim 2, wherein the solubilizing agent is selected from the group consisting of Tween 80, polyoxyethylene hydrogenated castor oil, polyethylene glycol 400, poloxamer 188, and polyoxyethylene hydrogenated stearin. One or more of acid esters, polyoxyethylene castor oil and its derivatives,

   Preferably, the solubilizer is polyoxyethylene hydrogenated castor oil;

   Preferably, the polyoxyethylene hydrogenated castor oil is selected from one or more of polyoxyethylene 40 hydrogenated castor oil, polyoxyethylene 54 hydrogenated castor oil, and polyoxyethylene 60 hydrogenated castor oil;

   Preferably, the solubilizing agent is Tween 80 and/or polyoxyethylene 40 hydrogenated castor oil,

   Preferably, the solubilizer is Tween 80 and polyoxyethylene 40 hydrogenated castor oil;

   The osmotic pressure regulator is selected from one or more of sodium chloride, potassium chloride, mannitol, glycerin, propylene glycol, dextrose, boric acid and borax, preferably sodium chloride and/or mannitol, and more Preferred are sodium chloride and mannitol;

   The bacteriostatic agent is selected from one or more of benzalkonium chloride, thimerosal, dumiphene, xanthanol, chlorobutanol, parabens, and sorbic acid, preferably benzalkonium chloride.
4. The pharmaceutical composition according to any one of claims 1 to 3, characterized in that the pharmaceutical composition includes a compound of formula I, Tween 80 and polyoxyethylene 40 hydrogenated castor oil;

   Or the pharmaceutical composition includes a compound of formula I and polyoxyethylene 40 hydrogenated castor oil;

   Or the pharmaceutical composition includes a compound of formula I, polyoxyethylene 40 hydrogenated castor oil, sodium chloride, mannitol and benzalkonium chloride;

   Alternatively, the pharmaceutical composition includes a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil and sodium chloride;

   Alternatively, the pharmaceutical composition includes a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, sodium chloride and benzalkonium chloride;

   Alternatively, the pharmaceutical composition includes a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, sodium chloride and mannitol;

   Alternatively, the pharmaceutical composition includes a compound of formula I, Tween 80, polyoxyethylene 40 hydrogenated castor oil, sodium chloride, mannitol and benzalkonium chloride.
5. The pharmaceutical composition according to any one of claims 1 to 4, characterized in that the pharmaceutical composition includes the following components by weight:
   
6. The pharmaceutical composition according to any one of claims 2 to 4, characterized in that the pharmaceutical composition includes the following components by weight:

   1 part of compound of formula I;

   Tween 80 0.3-3.5 parts;

   Polyoxyethylene 40 hydrogenated castor oil 2-35 parts;

   Alternatively, the pharmaceutical composition includes the following components by weight:
   

   Alternatively, the pharmaceutical composition includes the following components by weight:
   
   

   Alternatively, the pharmaceutical composition includes the following components by weight:
   

   Alternatively, the pharmaceutical composition includes the following components by weight:
   

   Alternatively, the pharmaceutical composition includes the following components by weight:
   

   Alternatively, the pharmaceutical composition includes the following components by weight:
   
7. The pharmaceutical composition according to any one of claims 1-6, wherein the drug The dosage form of the composition is a solution, suspension, emulsion, micelle or gel;

   Preferably, the content of the compound of formula I in the pharmaceutical composition is 0.01-2 mg/mL, preferably 0.05-1 mg/mL;

   Preferably, the crystal form of the compound of formula I is crystal form A or crystal form B;

   Preferably, the pH of the pharmaceutical composition is 4.0-6.0.
8. A method for preparing the pharmaceutical composition according to any one of claims 1 to 7, characterized in that the preparation method includes the following steps:

   The compound of formula I is mixed with a solubilizer, an osmotic pressure regulator and/or a bacteriostatic agent in corresponding parts by weight to obtain the pharmaceutical composition.
9. The preparation method according to claim 8, characterized in that the preparation method includes the following steps:

   (1) Dissolve the osmotic pressure regulator with water for injection to obtain solution I;

   (2) Mix the solubilizer and the compound of formula I evenly to obtain mixture II;

   (3) Mix solution I and mixture II and stir evenly to obtain solution III;

   (4) Dissolve the bacteriostatic agent in water for injection to obtain solution IV;

   (5) Mix the solution III and the solution IV, dilute to full volume with water for injection, and perform three-stage filtration and sterilization with a microporous filter membrane to obtain the pharmaceutical composition.
10. Application of the pharmaceutical composition according to any one of claims 1 to 7 or the pharmaceutical composition prepared by the preparation method according to claims 8 or 9 in the preparation of medicines for preventing and/or treating glaucoma or ocular hypertension .