EA045716B1 - COMPOSITION OF CONDENSED TRICYCLIC GAMA-AMINO ACID DERIVATIVES FOR TREATMENT AND/OR PREVENTION OF MECHANICAL PAIN AND ITS PREPARATION - Google Patents

Description

The present invention relates to the field of medicine, in particular to a pharmaceutical composition of condensed tricyclic gamma-amino acid derivatives and its use.

Prior Art

Diabetic peripheral neuropathy (DPNP) is a common chronic complication of diabetes mellitus. At least 25% of diabetic patients have suffered from painful diabetic peripheral neuropathy, and 50% of diabetic patients with more than a 25-year history of the disease develop neuralgia. There are currently more than 10 million patients with DPNP in China.

Postherpetic neuralgia (PHN) is the most common complication of herpes zoster. The annual incidence of herpes zoster is about 3-5%, and about 9-34% of patients with herpes zoster develop PHN. The incidence of herpes zoster tends to increase with age. About 65% of patients with herpes zoster aged 60 years and older develop PHN, and the number of patients aged 70 years and older reaches 75%. There are no relevant studies available in China. However, according to the above data, it is estimated that there are about 4 million patients with PHN in China.

Fibromyalgia syndrome (MF) is a group of clinical syndromes of unknown etiology and is characterized by systemic pain and significant physical discomfort. The overall prevalence of FS is 2.7% worldwide, 2% in the United States, and 0.8% in China.

At present, drugs for the symptomatic treatment of peripheral neuralgia mainly include anticonvulsants, tricyclic antidepressants, opioid analgesics, topical analgesics and the like.

FDA-approved therapeutic agents include voltage-gated calcium channel antagonists such as pregabalin, gabapentin, 5-HT/NE reuptake inhibitors such as duloxetine, milnacipran, μ-opioid receptor agonist/NE reuptake inhibitor , such as tapentadol, and topical medications such as lidocaine patch, capsaicin patch. Currently, the CFDA approves only pregabalin and gabapentin for the treatment of PHN, while no drugs have been approved for the treatment of DPNP and SF.

Voltage-gated sodium channels consist of the α1 subunit and auxiliary α2δ-, β-, γ protein subunits. The α2δ protein can regulate calcium channel density and voltage-dependent calcium channel dynamics (Felix et al. (1997) J. Neuroscience 17: 6884-6891; Klugbauer et al. (1999) J. Neuroscience 19: 684-691; Hobom et al. (2000) Eur J Neuroscience 12: 1217-1226 and Qin et al (2002) Mol Pharmacol 62: 485496). Compounds that exhibit high binding affinity to the α2δ subunit of the voltage-gated calcium channel, such as pregabalin and gabapentin, have been demonstrated to be effective in the treatment of pain. In mammals, the α2δ protein has 4 subtypes, each of which is encoded by a separate gene. α2δ subtype 1 and subtype 2 show high affinity for pregabalin, whereas α2δ subtype 3 and subtype 4 do not have significant drug binding capacity.

Clinically, pregabalin is prescribed three times a day or twice a day when clinical compliance of patients is low. Pregabalin only has a pain-relieving effect in approximately 30-50% of patients and has little effect on refractory forms of DPPN and the like. Side effects such as dizziness, drowsiness, weight gain, edema and the like frequently occur during clinical use, which may affect the quality of life of patients.

Summary of the invention

The present invention provides a pharmaceutical composition of a condensed tricyclic gamma-amino acid derivative having a novel structure and good efficacy, or a pharmaceutically acceptable salt thereof, and its use in the field of analgesia.

The present invention also provides a method of treating and/or preventing pain, comprising administering an effective amount of a condensed tricyclic gamma-amino acid derivative or a pharmaceutically acceptable salt thereof.

The novel compound, a condensed tricyclic gamma-amino acid derivative (structural formula (I)) or a pharmaceutically acceptable salt thereof, provided in the present invention is an α2δ-subunit calcium channel antagonist that can competitively bind to the α2δ-subunit of calcium channels. A compound that corresponds to the endogenous inhibitory neurotransmitter, GABA (gamma-aminobutyric acid), associated with the regulation of brain neuronal activity, has a therapeutic effect on peripheral neuropathy caused by diabetes mellitus, postherpetic neuralgia and fibromyalgia, and is thus expected to overcome the deficiency pregabalin.

According to the present invention, a pharmaceutical composition is provided containing:

(1) an active substance having the structure represented by formula (I) or a pharmaceutically acceptable salt thereof in an amount of 1 to 45% by weight;

- 1 045716 (2) filler in an amount from 50 to 95% by weight, containing mannitol in an amount from 9 to 35% by weight, microcrystalline cellulose A in an amount from 3 to 60% by weight and microcrystalline cellulose B in an amount from 17 to 72% by weight, wherein microcrystalline cellulose A is selected from the group consisting of microcrystalline cellulose PH14, microcrystalline cellulose PH12, microcrystalline cellulose 12 and microcrystalline cellulose 14; microcrystalline cellulose B is selected from the group consisting of microcrystalline celluloses PH102, PH105, PH103, PH301, PH101, PH112, PH302 and microcrystalline celluloses 102, 105, 103, 301, 101, 112, 302;

(3) one or more lubricants in an amount of from 0.1 to 5.5% by weight;

where the sum of the mass percentages of all components is 100%;

where formula (I) has the following structure with ₂ n (I).

According to the present invention, wherein formula (I) has the following structure b with ₂ H

I (II).

In one embodiment, a pharmaceutically acceptable salt of a compound of formula (I) has the following:

structure

In one embodiment, optionally, a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 1-100 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 2-80 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 2-60 mg.

Optionally, the compound of formula (II) or a pharmaceutically acceptable salt thereof is included in an amount of 2-50 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 5-50 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 5-40 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 5-30 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 5-20 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 5-10 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 10-80 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 10-60 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 10-50 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 20-80 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 20-60 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 20-50 mg.

Optionally, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 30-50 mg.

In a specific embodiment, a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is included in an amount of 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg or 50 mg.

In any of the above embodiments, the pharmaceutically acceptable salt is selected from benzenesulfonate.

- 2 045716

Also provided is a method of treating and/or preventing pain, comprising administering an effective amount of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof, where the effective amount is possibly in the range from 1 to 100 mg, or from 2 to 80 mg, or from 2 to 60 mg, or from 2 to 50 mg, or from 5 to 50 mg, or from 5 to 40 mg, or from 5 to 30 mg, or from 5 to 20 mg, or from 10 to mg, or from 10 up to 70 mg, or from 10 to 60 mg, or from 10 to 50 mg, or from 10 to 45 mg, or from 15 to 70 mg, or from 15 to 60 mg, or from 15 to 50 mg, or from 15 to 45 mg, or from 20 to 70 mg, or from 20 to 60 mg, or from 20 to 50 mg, or from 20 to 45 mg.

Preferably, the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is a benzene sulfonate thereof.

When a pharmaceutically acceptable salt is administered as an active ingredient, the effective dose is based on the free base.

In one embodiment, administration is orally.

In one embodiment, the pain includes: postherpetic neuralgia, trigeminal neuralgia, migraine, pain due to osteoarthritis or articular rheumatism, low back pain, sciatic neuralgia, dental pain, pain due to burns, pain due to diabetic neuropathy, pain due to chemotherapy-induced neuropathy , HIV-associated neuralgia, AIDS-associated neuralgia, cancer neuralgia or non-neuropathic pain, acute or chronic tension headache, post-operative pain or fibromyalgia. In a particular embodiment, the pain includes: stherpetic neuralgia, diabetic neuropathy pain, or fibromyalgia.

It is also proposed to use the above pharmaceutical composition in the preparation of a medicinal agent for the treatment and/or prevention of pain. Preferably the pain includes:

postherpetic neuralgia, trigeminal neuralgia, migraine, pain due to osteoarthritis or joint form of rheumatism, low back pain, sciatic neuralgia, toothache, pain caused by burns, pain due to diabetic neuropathy, pain due to neuropathy caused by chemotherapy, HIV-associated neuralgia, AIDS- associated neuralgia, neuralgia due to cancer or non-neuropathic pain, acute or chronic tension-type headache, postoperative pain or fibromyalgia.

Also proposed is the use of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof in the preparation of a medicinal product for the treatment and/or prevention of pain.

In one embodiment, optionally, a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof is administered in a single dose of 1-100 mg.

Possibly a compound of formula (I) in a single dose of 2-80 mg.

Possibly a compound of formula (I) in a single dose of 2-60 mg.

Possibly a compound of formula (I) in a single dose of 2-50 mg.

Possibly a compound of formula (I) in a single dose of 5-50 mg.

Possibly a compound of formula (I) in a single dose of 5-40 mg.

Possibly a compound of formula (I) in a single dose of 5-30 mg.

Possibly a compound of formula (I) in a single dose of 5-20 mg.

Possibly a compound of formula (I) in a single dose of 5-10 mg.

Possibly a compound of formula (I) in a single dose of 10-80 mg.

Possibly a compound of formula (I) in a single dose of 10-60 mg.

Possibly a compound of formula (I) in a single dose of 10-50 mg.

Possibly a compound of formula (I) in a single dose of 20-80 mg.

Possibly a compound of formula (I) in a single dose of 20-60 mg.

Possibly a compound of formula (I) in a single dose of 20-50 mg.

Possibly a compound of formula (I) in a single dose of 30-50 mg.

or or or or or or or or or or or or (II) (II) (II) (II) (II) (II) (II) (II) (II) (II) (II) (II ) (II) (II) (II) or or or or or or or or or or or or his his his his his his his his his his his his his pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable pharmaceutically acceptable salt salt salt salt salt salt salt salt salt salt salt salt salt salt salt introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced introduced

In a specific embodiment, a compound of formula (I) or (II) or a pharmaceutically acceptable

- 3 045716 salt included in amounts of 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg or 50 mg.

In each of the above embodiments or uses, the pharmaceutically acceptable salt is selected from benzenesulfonates.

In each of the above embodiments or uses, when a pharmaceutically acceptable salt of a compound of formula (I) or (II) is used in the preparation of a medicament for the treatment and/or prevention of pain, the unit dose is the value in terms of the free base.

Another object of the present invention is to improve the stability of the drug and solve the problem of uniformity of composition in the production of the drug, and thereby obtain a drug having adequate uniformity of composition and good chemical stability. According to the present invention, there is provided a preparation containing a compound of formula (I) or formula (II) as an active ingredient, which may be in the form of tablets, capsules, granules and the like.

The drug proposed in the present invention dissolves quickly, has good chemical stability, is safe, effective and has few side effects.

According to the present invention, a pharmaceutical composition is provided, which, based on the total weight of the pharmaceutical composition (100%), contains:

1) an active substance having the structure represented by formula (I) or formula (II), or a pharmaceutically acceptable salt thereof in an amount of 1 to 45% by weight, or in an amount of 1 to 40% by weight in some embodiments, or in in an amount from 1 to 35% by weight in some embodiments, or in an amount from 1 to 30% by weight in some embodiments, or in an amount from 1 to 25% by weight in some embodiments, or in an amount from 5 to 45% by weight in some embodiments, or in an amount from 5 to 40% by weight in some embodiments, or in an amount from 5 to 35% by weight in some embodiments, or in an amount from 5% to 30% by weight in some embodiments, or in an amount of from 5 to 25% by weight in some embodiments, or in an amount from 5 to 21% by weight in some embodiments;

2) optionally one or more fillers in an amount from 50 to 95% by weight, or in an amount from 55 to 94% by weight in some embodiments, or in an amount from 79 to 94% by weight in some embodiments;

3) optionally one or more lubricants in an amount of 0.1 to 6% by weight, or in an amount of 0.1 to 5.5% by weight in some embodiments, or in an amount of 0.1 to 2% by weight in some embodiments, or in an amount from 0.1 to 1.5% by weight in some embodiments, or in an amount from 0.5 to 1.5% by weight in some embodiments, or in an amount from 0.5 to 1% by weight in some embodiments, or in an amount from 0.5 to 0.9% by weight in some embodiments, or in an amount from 0.5 to 0.8% by weight in some embodiments, or in an amount from 0. 5 to 0.7% by weight in some embodiments, or in an amount of 0.5 to 0.6% by weight in some embodiments;

wherein the above ranges may be combined arbitrarily, provided that the sum of the weight percentages of all components is 100%; and wherein the structure of formula (I) is as follows:

H / ^NH 2 ΐΓ^ΟΟ ₂ Η \i/ ₍₁₎

The pharmaceutical composition according to the present invention, wherein the structure of formula (I) is as follows:

Pharmaceutical composition according to the present invention, wherein the pharmaceutically acceptable salt of the compound of formula (I) has the following structure: nh ₂ n S

1(^CO ₂ H O _pn

The pharmaceutically acceptable salt of the compound of formula (I) has a crystalline structure and characteristic diffraction peaks at 2Θ angles of 9.72° ± 0.2°; 14.00°±0.2°; 16.33°±0.2°; 19.32°±0.2°; 20.46°±0.2°; 21.69°±0.2° and 25.33°±0.2° in the X-ray powder diffraction spectrum using Cu-Ka radiation. It also has characteristic diffraction peaks at 2Θ angles of 11.21° ± 0.2°; 15.16°±0.2°; 18.87°±0.2°; 19.88°±0.2°; 23.47°±0.2° and 27.96°±0.2° in the X-ray diffraction spectrum

- 4 045716 on powder using Cu-Ka radiation. It also has characteristic diffraction peaks at angles 2Θ 21.30°±0.2°; 25.40°±0.2°; 29.82°±0.2° in the X-ray powder diffraction spectrum. In addition, a crystal of a pharmaceutically acceptable salt of the compound of formula (I) has an X-ray powder diffraction spectrum using Cu-Ka radiation as shown in FIG. 4. Moreover, in FIG. 5 shows a TGA/DSC (thermogravimetric analysis/differential scanning calorimetry) spectrum of a crystal of a pharmaceutically acceptable salt of the compound of formula (I). Moreover, a pharmaceutically acceptable salt of the compound of formula (I) has a single crystal diffraction spectrum as shown in FIG. 6, wherein the pharmaceutically acceptable salt of the compound of formula (I) has the following structure:

The composition of the present invention may be a pharmaceutical preparation in the form of tablets, granules, microcapsules, pills or capsules. In some embodiments, it is in the form of tablets or capsules. Additionally, in some embodiments it is in the form of capsules.

The composition according to the present invention contains the active substance in an amount of from 1 to 45% by weight, or in some embodiments in an amount from 1 to 40% by weight, or in some embodiments in an amount from 1 to 35% by weight, or in some embodiments in in an amount of 1 to 30% by weight, or in some embodiments in an amount of 1 to 25% by weight, or in some embodiments in an amount of 5 to 45% by weight, or in some embodiments in an amount of 5 to 40% by weight , or in some embodiments in an amount from 5 to 35% by weight, or in some embodiments in an amount from 5 to 30% by weight, or in some embodiments in an amount from 5 to 25% by weight, or in some embodiments in an amount from 5 to 21% by weight, or in some embodiments in an amount from 5.5 to 21% by weight, or in some embodiments in an amount from 5.5 to 20.5% by weight, or in some embodiments in an amount from 5. 5 to 20% by weight.

The filler in the composition of the present invention is one or more substances selected from mannitol, low-substituted hydroxypropylcellulose and microcrystalline cellulose. In some embodiments, the filler is selected from one or more of mannitol and microcrystalline cellulose. In some embodiments, it is a combination of mannitol and microcrystalline cellulose.

The filler in the composition of the present invention is selected from a combination of mannitol and microcrystalline cellulose with a mannitol content of from 9 to 40% by weight. In some embodiments, the mannitol content ranges from 9 to 35% by weight; in some embodiments, the mannitol content is from 10 to 28% by weight; in some embodiments, the mannitol content is from 9 to 25% by weight; in some embodiments, the mannitol content is from 9 to 20% by weight; in some embodiments, the mannitol content is from 15 to 35% by weight; in some embodiments, the mannitol content is from 15 to 30% by weight; in some embodiments, the mannitol content is from 15 to 28% by weight; in some embodiments, the mannitol content is from 15 to 25% by weight; in some embodiments, the mannitol content is from 15 to 20% by weight.

The filler in the composition of the present invention is selected from a combination of mannitol and microcrystalline cellulose, wherein said microcrystalline cellulose comprises microcrystalline cellulose A and microcrystalline cellulose B. Microcrystalline cellulose A is selected from microcrystalline cellulose PH14, microcrystalline cellulose PH12, microcrystalline cellulose 12, or microcrystalline cellulose 14. In some in embodiments, microcrystalline cellulose A is selected from microcrystalline cellulose 12; in some embodiments, microcrystalline cellulose A is selected from microcrystalline cellulose PH12; in some embodiments, microcrystalline cellulose A is selected from microcrystalline cellulose PH14; in some embodiments, microcrystalline cellulose A is selected from microcrystalline cellulose 14. Microcrystalline cellulose B is selected from microcrystalline cellulose PH102, PH105, PH103, PH301, PH101, PH112, PH302 and microcrystalline cellulose 102, 105, 103, 301, 101, 112, 302.V In some embodiments, microcrystalline cellulose B is selected from microcrystalline cellulose 102; in some embodiments, microcrystalline cellulose B is selected from microcrystalline cellulose PH102.

The filler in the composition according to the present invention is selected from a combination of mannitol and microcrystalline cellulose, wherein said microcrystalline cellulose contains microcrystalline cellulose A and B, which are respectively selected from microcrystalline cellulose 102 and microcrystalline cellulose 12.

The filler in the composition according to the present invention is selected from a combination of mannitol and microcrystalline cellulose, where the microcrystalline cellulose contains microcrystalline cellulose A and B with a content of microcrystalline cellulose A from 3 to 60% by weight. In no

- 5 045716 in which embodiments the content of microcrystalline cellulose A is from 3 to 59% by weight; in some embodiments, the microcrystalline cellulose A content ranges from 3 to 58% by weight; in some embodiments, the microcrystalline cellulose A content is from 3 to 10% by weight; in some embodiments, the microcrystalline cellulose A content is from 3 to 8% by weight; in some embodiments, the microcrystalline cellulose A content is from 5 to 15% by weight; in some embodiments, the microcrystalline cellulose A content is from 5 to 12% by weight; in some embodiments, the microcrystalline cellulose A content is from 5 to 10% by weight; in some embodiments, the microcrystalline cellulose A content is from 5 to 9% by weight; in some embodiments, the microcrystalline cellulose A content is from 5 to 8% by weight; in some embodiments, the microcrystalline cellulose A content is from 5 to 7.5% by weight; in some embodiments, the microcrystalline cellulose A content is from 5.5 to 7.5% by weight.

The filler in the composition of the present invention is selected from a combination of mannitol and microcrystalline cellulose, where the microcrystalline cellulose contains microcrystalline cellulose A and B with a microcrystalline cellulose B content of from 17 to 80% by weight. In some embodiments, the microcrystalline cellulose B content ranges from 17 to 72% by weight; in some embodiments, the microcrystalline cellulose B content is from 40 to 80% by weight; in some embodiments, the content of microcrystalline cellulose B is from 40 to 70% by weight; in some embodiments, the microcrystalline cellulose B content is from 40 to 75% by weight; in some embodiments, the content of microcrystalline cellulose B is from 41 to 72% by weight; in some embodiments, the microcrystalline cellulose B content is from 42 to 72% by weight; in some embodiments, the content of microcrystalline cellulose B is from 50 to 80% by weight; in some embodiments, the microcrystalline cellulose B content is from 50 to 75% by weight; in some embodiments, the microcrystalline cellulose B content is from 50 to 70% by weight; in some embodiments, the content of microcrystalline cellulose B is from 55 to 80% by weight; in some embodiments, the content of microcrystalline cellulose B is from 55 to 75% by weight; in some embodiments, the microcrystalline cellulose B content is from 56 to 72% by weight; in some embodiments, the microcrystalline cellulose B content is from 56 to 61% by weight.

The filler in the composition of the present invention is selected from a combination of mannitol and microcrystalline cellulose, where the microcrystalline cellulose contains microcrystalline cellulose A and B with a microcrystalline cellulose A content of 5 to 8% by weight. In some embodiments, the microcrystalline cellulose A content is from 5 to 7.5% by weight; in some embodiments, the microcrystalline cellulose A content is from 5.5 to 7.5% by weight, and the microcrystalline cellulose B content is from 55 to 75% by weight. In some embodiments, the microcrystalline cellulose B content is from 56 to 72% by weight; in some embodiments, the microcrystalline cellulose B content is from 56 to 61% by weight.

The filler in the composition according to the invention is selected from a combination of mannitol and microcrystalline cellulose, where the microcrystalline cellulose contains microcrystalline cellulose 12 and microcrystalline cellulose 102 with a content of microcrystalline cellulose 12 from 5 to 8% by weight. In some embodiments, the content of microcrystalline cellulose 12 is from 5 to 7.5% by weight; in some embodiments, the content of microcrystalline cellulose 12 is from 5.5 to 7.5% by weight, and the content of microcrystalline cellulose 102 is from 55 to 75% by weight. In some embodiments, the content of microcrystalline cellulose 102 is from 56 to 72% by weight; in some embodiments, the content of microcrystalline cellulose 102 is from 56 to 61% by weight.

Microcrystalline cellulose 12 has a D50 particle size ranging from 130 μm to 230 μm, and a D90 particle size ranging from 270 μm to 450 μm. Microcrystalline cellulose 102 has a D50 particle size ranging from 90 μm to 150 μm, and a D90 particle size ranging from 190 μm to 300 μm.

The filler in the composition according to the invention is selected from a combination of mannitol and microcrystalline cellulose, where the microcrystalline cellulose contains microcrystalline cellulose 12 and microcrystalline cellulose 102 with a content of microcrystalline cellulose 12 from 5 to 8% by weight. In some embodiments, the content of microcrystalline cellulose 12 is from 5 to 7.5% by weight; in some embodiments, the content of microcrystalline cellulose 12 is from 5.5 to 7.5% by weight; and the content of microcrystalline cellulose 102 is from 55% to 75% by weight. In some embodiments, the content of microcrystalline cellulose 102 is from 56% to 72% by weight; in some embodiments, the content of microcrystalline cellulose 102 is from 56 to 61% by weight. Microcrystalline cellulose 12 has a D50 particle size ranging from 130 μm to 230 μm, and a D90 particle size ranging from 270 μm to 450 μm. Microcrystalline cellulose 102 has a D50 particle size ranging from 90 μm to 150 μm, and a D90 particle size ranging from 190 μm to 300 μm.

- 6 045716

The filler in the composition of the present invention is selected from a combination of mannitol and microcrystalline cellulose, wherein the weight percentage of filler in the composition is the sum of the microcrystalline cellulose content containing microcrystalline cellulose A, B, and mannitol content, the sum of which and the sum of the other ingredients together is 100%.

The filler in the composition according to the present invention is selected from a combination of mannitol and microcrystalline cellulose, where the microcrystalline cellulose has a D50 particle size ranging from 90 μm to 230 μm. When microcrystalline cellulose has a D50 particle size of less than 90 µm, a binder should be added to the composition.

The lubricant in the composition according to the present invention is one or more than one selected from stearic acid, magnesium stearate, sodium stearyl fumarate and glycerol behenate. In some embodiments, it is selected from magnesium stearate or stearic acid; in some embodiments it is magnesium stearate.

The lubricant is included in the composition of the present invention in an amount of from 0.5 to 6% by weight, or in some embodiments in an amount from 0.5 to 5.5% by weight, or in some embodiments in an amount from 0.5 to 5 .3% by weight, or in some embodiments from 0.5 to 2% by weight, or in some embodiments from 0.5 to 1.5% by weight, or in some embodiments from 0.5% by weight to 1% by weight, or in some embodiments in an amount from 0.5 to 0.9% by weight, or in some embodiments in an amount from 0.5 to 0.8% by weight, or in some embodiments in an amount from 0 .5 to 0.7% by weight, or in some embodiments from 0.5 to 0.6% by weight, or in some embodiments from 0.55 to 0.6% by weight, or in some embodiments in an amount from 0.51 to 0.55% by weight.

The composition according to the present invention may further optionally contain one or more than one binder, if necessary.

The binder may be selected from hydroxypropylcellulose or povidone.

The binder is included in an amount of 1 to 10% by weight, or in some embodiments in an amount of 1 to 6%, or in some embodiments in an amount of 1 to 5%, or in some embodiments in an amount of 1 to 4%, or in some embodiments in an amount of 1 to 3%, or in some embodiments in an amount of 4%, or in some embodiments in an amount of 3%.

The composition of the present invention may further optionally contain one or more disintegrants, if desired.

The leavening agent may be selected from croscarmellose sodium and sodium carboxymethyl starch.

The disintegrant is included in an amount of 1 to 10% by weight, or in some embodiments in an amount of 1 to 5%, or in some embodiments in an amount of 1 to 4%, or in some embodiments in an amount of 3 to 4%, or in some embodiments in the amount of 4%.

The composition according to the present invention may further optionally contain one or more glidants, if necessary.

The glidant is selected from silica or talc.

The glidant is included in an amount of 1 to 10% by weight, or in some embodiments in an amount of 1 to 6%, or in some embodiments in an amount of 1 to 5%, or in some embodiments in an amount of 1 to 4%, or in some embodiments in an amount of 1 to 3%, or in some embodiments in an amount of 4%, or in some embodiments in an amount of 3%.

The above percentages are based on the total weight of the composition (100%), however, certain ranges of percentages of any individual component may be arbitrarily combined with the percentages of other component(s) provided that the total percentage is 100%.

The composition according to the present invention is a capsule preparation containing:

1) a pharmaceutically acceptable salt of a compound of formula (I) as the active substance, which is included in an amount of from 1 to 45% by weight, or in some embodiments in an amount from 1 to 40% by weight, or in some embodiments in an amount from 1 to 35% by weight, or in some embodiments from 1 to 30% by weight, or in some embodiments from 1 to 25% by weight, or in some embodiments from 5 to 45% by weight, or in some embodiments in an amount from 5 to 40% by weight, or in some embodiments in an amount from 5 to 35% by weight, or in some embodiments in an amount from 5 to 30% by weight, or in some embodiments in an amount from 5 to 25% by weight, or in some embodiments in an amount from 5 to 21% by weight, or in some embodiments in an amount from 5.5 to 21% by weight, or in some embodiments in an amount from 5.5 to 20.5% by weight , or in some embodiments in an amount of from 5.5 to 20% by weight;

2) a filler containing mannitol, microcrystalline cellulose A and microcrystalline cellulose B, where mannitol is included in an amount of from 9 to 40% by weight, or in some embodiments in an amount from 9 to 35% by weight, or in some embodiments in an amount from 10 up to 28% by weight, or in some embodiments from 9 to 25% by weight, or in some embodiments from 9 to 20% by weight, or in some embodiments from 15 to 35% by weight, or in

- 7 045716 some embodiments in an amount from 15 to 30% by weight, or in some embodiments in an amount from 15 to 28% by weight, or in some embodiments in an amount from 15 to 25% by weight, or in some embodiments in an amount from 15 to 20% by weight; microcrystalline cellulose A is included in an amount from 3 to 60% by weight, or in some embodiments in an amount from 3 to 59% by weight, or in some embodiments in an amount from 3 to 58% by weight, or in some embodiments in an amount from 3 up to 10% by weight, or in some embodiments in an amount from 3 to 8% by weight, or in some embodiments in an amount from 5 to 15% by weight, or in some embodiments in an amount from 5 to 12% by weight, or in in some embodiments in an amount from 5 to 10% by weight, or in some embodiments in an amount from 5 to 9% by weight, or in some embodiments in an amount from 5 to 8% by weight, or in some embodiments in an amount from 5 to 7% .5% by weight, or in some embodiments in an amount of from 5.5 to 7.5% by weight; and microcrystalline cellulose B is included in an amount from 17 to 80% by weight, or in some embodiments in an amount from 40 to 80% by weight, or in some embodiments in an amount from 40 to 70% by weight, or in some embodiments in an amount from 40 to 75% by weight, or in some embodiments from 41 to 72% by weight, or in some embodiments from 42 to 72% by weight, or in some embodiments from 50 to 80% by weight, or in some embodiments in an amount from 50 to 75% by weight, or in some embodiments in an amount from 50 to 70% by weight, or in some embodiments in an amount from 55% to 80% by weight, or in some embodiments in an amount from 55 up to 75% by weight, or in some embodiments in an amount from 56 to 72% by weight, or in some embodiments in an amount from 56 to 61% by weight;

3) magnesium stearate as a lubricant, which is included in an amount from 0.5 to 6% by weight, or in some embodiments in an amount from 0.5 to 5.5% by weight, or in some embodiments in an amount from 0. 5 to 5.3% by weight, or in some embodiments from 0.5 to 2% by weight, or in some embodiments from 0.5 to 1.5% by weight, or in some embodiments from 0.5 to 1% by weight, or in some embodiments from 0.5 to 0.9% by weight, or in some embodiments from 0.5 to 0.8% by weight, or in some embodiments from in an amount of 0.5 to 0.7% by weight, or in some embodiments in an amount of 0.5 to 0.6% by weight, or in some embodiments in an amount of 0.55 to 0.6% by weight, or in some embodiments, in an amount of from 0.51 to 0.55% by weight;

wherein certain ranges of percentages of any individual component may be arbitrarily combined with percentages of other component(s) so long as the total percentage of all components is 100%; and wherein the pharmaceutically acceptable salt of the compound of formula (I) has the following structure:

The composition according to the present invention may further contain a glidant selected from silica and talc. In some embodiments, the glidant is silica.

The compositions of the present invention may further contain a binder selected from hydroxypropylcellulose or povidone. In some embodiments, the binder is hydroxypropylcellulose. In particular, when the composition contains only one excipient, the composition further contains a binder.

The present invention also relates to a pharmaceutical composition in tablet form containing:

1) a pharmaceutically acceptable salt of a compound of formula (I) as the active substance, which is included in an amount of from 4 to 35% by weight, or in some embodiments in an amount of 4% by weight, or in some embodiments in an amount of 11% by weight, or in some embodiments in an amount of 20% by weight, or in some embodiments in an amount of 34% by weight;

2) a filler containing mannitol and microcrystalline cellulose B, wherein mannitol is included in an amount from 13 to 60% by weight, or in some embodiments in an amount from 13 to 40% by weight, or in some embodiments in an amount from 13 to 20% by weight weight; and microcrystalline cellulose B is included in an amount from 10 to 60% by weight, or in some embodiments in an amount from 19 to 60% by weight, or in some embodiments in an amount from 19 to 50% by weight, or in some embodiments in an amount from 19 to 40% by weight, or in some embodiments from 40 to 60% by weight, or in some embodiments from 40 to 50% by weight, or in some embodiments from 50 to 60% by weight;

3) a binder selected from hydroxypropyl methylcellulose, povidone or hydroxypropylcellulose, in an amount of 4 to 10% by weight; in some embodiments selected from hydro

- 8 045716 xypropyl methylcellulose, in some embodiments in an amount of 4 to 7%, or in some embodiments in an amount of 5 to 7%, or in some embodiments in an amount of 5%, or in some embodiments in an amount of 6%, or in some embodiments in the amount of 7%;

4) a disintegrant selected from croscarmellose sodium, croscarmellose calcium, crospovidone or sodium carboxymethyl starch, in an amount of from 2 to 10% by weight; in some embodiments selected from croscarmellose sodium, in some embodiments in an amount of 6 to 10% by weight, or in some embodiments in an amount of 6 to 9% by weight, or in some embodiments in an amount of 6% by weight, or in some embodiments in an amount of 7% by weight, or in some embodiments in an amount of 8% by weight, or in some embodiments in an amount of 9% by weight, or in some embodiments in an amount of 10% by weight;

5) a lubricant selected from magnesium stearate and sodium stearyl fumarate, in an amount of 0.5 to 1% by weight; in some embodiments selected from magnesium stearate, or in some embodiments in an amount of from 0.6 to 1% by weight, or in some embodiments in an amount of 0.6% by weight, or in some embodiments in an amount of 0.7% by weight, or in some embodiments in an amount of 0.8% by weight, or in some embodiments in an amount of 0.9% by weight, or in some embodiments in an amount of 1% by weight;

wherein certain ranges of percentages of any individual component may be arbitrarily combined with percentages of other component(s) so long as the total percentage of the composition is 100%; and wherein the pharmaceutically acceptable salt of the compound of formula (I) has the following structure:

In the composition of the present invention, which is a preparation in the form of tablets or capsules, the active substance is present, based on the free base, in an amount of from 1 to 100 mg, or in some embodiments in an amount from 2 to 80 mg, or in some embodiments in an amount of 2 to 60 mg, or in some embodiments in an amount of 2 to 50 mg, or in some embodiments in an amount of 5 to 50 mg, or in some embodiments in an amount of 5 to 40 mg, or in some embodiments in an amount of from 5 to 30 mg, or in some embodiments from 5 to 20 mg, or in some embodiments from 5 to 10 mg, or in some embodiments from 10 to 80 mg, or in some embodiments from 10 up to 60 mg, or in some embodiments in an amount from 10 to 50 mg, or in some embodiments in an amount from 20 to 80 mg, or in some embodiments in an amount from 20 to 60 mg, or in some embodiments in an amount from 20 to 50 mg, or in some embodiments in an amount of 30 to 50 mg.

In the composition according to the present invention, which is a preparation in the form of tablets or capsules, the active substance is present, in terms of the free base, in an amount of 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg or 50 mg, or in some embodiments in an amount of 5 mg, 10 mg or 20 mg, or in some embodiments in an amount of 5 mg or 20 mg.

The present invention also provides a method for preparing a pharmaceutical composition, comprising:

1) passing the active substances through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use;

2) adding the sifted active substance and filler to the mixer with multidirectional movement for uniform mixing;

3) adding lubricant to the multi-directional mixer to mix with the mixture obtained in step (2);

or, alternatively, when a disintegrant and/or a glidant are included in the mixture, adding the lubricant, disintegrant and glidant to a multi-directional mixer to mix with the mixture obtained in step (2); And

4) filling the capsule with the mixture obtained in step (3).

The present invention also provides a method for preparing a pharmaceutical composition, comprising:

1) passing the active substances through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use;

2) preparing a 40% aqueous solution of ethanol as a solvent;

3) adding the sieved active substance and filler with the binder to the wet granulator and adding the solvent obtained in step (2) for shear mixing to obtain a soft material;

4) passing soft material through a sieve to obtain particles, and performing static drying

- 9 045716 ki or dynamic drying in a fluidized bed;

5) granulating the dried soft material and then adding it with a glidant and disintegrant to a multi-directional mixer for uniform mixing;

6) adding lubricant to the multi-directional mixer for mixing; And

7) filling the capsule with the mixture obtained in step (6). Or alternatively, steps (2) and (3) can be replaced by:

2- 1) preparation of a binder and a 40% aqueous solution of ethanol in the form of a solution of the binder;

3- 1) adding the active substance and filler to the wet granulator and adding the binder solution obtained in step (2-1) for shear mixing to obtain a soft material.

The present invention also provides a method for preparing a pharmaceutical composition in tablet form, comprising:

1) passing the active ingredients through a 100 mesh sieve and passing the filler and disintegrant through a 60 mesh sieve;

2) adding the sieved active ingredient, filler and disintegrant into the high-speed wet granulator to ensure uniform mixing;

3) adding a binding solvent to the mixed powder obtained in step (2) to obtain a soft material;

4) passing the soft material through a 20 mesh screen to obtain particles, then subjecting it to static drying or dynamic fluid bed drying to reduce the moisture content to less than 2%;

5) passing the dried particles through a 24 mesh sieve for granulation, adding lubricant granules to the mixer with multi-directional motion to ensure uniform mixing;

6) pressing the mixture obtained in step (5) into tablet form. Or alternatively, steps (3) and (4) can be replaced by:

(3-1) adding the mixed powder obtained in step (2) to the fluidized bed, supplying the binder solution to the fluidized bed by a peristaltic pump, and producing particles by overhead spraying.

The present invention also relates to a method of treating and/or preventing pain, comprising: administering an effective dose of a compound of formula (I) or a pharmaceutically acceptable salt thereof as an active substance, wherein the effective dose is from 1 to 100 mg based on the free salt of the compound of formula ( I) or a pharmaceutically acceptable salt thereof, or in some embodiments 5 to 20 mg, or in some embodiments 5 mg or 20 mg. Pain includes: postherpetic neuralgia, trigeminal neuralgia, migraine, pain due to osteoarthritis or joint form of rheumatism, low back pain, sciatic neuralgia, dental pain, pain due to burns, pain due to diabetic neuropathy, pain due to chemotherapy-induced neuropathy, HIV-associated neuralgia, AIDS-associated neuralgia, cancer-related neuralgia or non-neuropathic pain, acute or chronic tension-type headache, postoperative pain or fibromyalgia; in some embodiments, the pain includes postherpetic neuralgia, diabetic neuropathy pain, or fibromyalgia.

The route of administration for the treatment method of the present invention is oral administration.

The present invention also relates to the use of a pharmaceutical composition in the manufacture of a medicament for the treatment and/or prevention of pain. Pain includes: postherpetic neuralgia, trigeminal neuralgia, migraine, pain due to osteoarthritis or articular rheumatism, low back pain, sciatic neuralgia, toothache, burn pain, pain due to diabetic neuropathy, pain due to chemotherapy-induced neuropathy, HIV-associated neuralgia, AIDS-associated neuralgia, neuralgia due to cancer or non-neuropathic pain, acute or chronic tension-type headache, postoperative pain or fibromyalgia; in some embodiments, the pain includes postherpetic neuralgia, diabetic neuropathy pain, or fibromyalgia.

Unless otherwise specified, the terms used in the specification and claims have the following meanings.

The term effective dose refers to the amount of a compound that produces, as appropriate, a physiological or medical response in a tissue, system, or subject, including an amount of the compound that, when administered to a subject, is sufficient to prevent the occurrence of or to reduce to some extent one or more than one symptom condition or disorder being treated.

In the absence of specific instructions, the expression mass percent in the present invention means

- 10 045716 refers to the weight percentage of a component based on the total weight of the composition, and the weight percent of each component of a formulation, such as a tablet or capsule, refers to the weight percentage of such component based on all components in the formulation, such as the tablet or capsule filler (including the active agents and excipients).

The term active agent in the present invention refers to the compound of formula (I) presented below:

or a compound of formula (II) presented below:

or a pharmaceutically acceptable salt of a compound of formula (I) or (II), such as:

Brief description of graphic materials

In fig. 1 is a graph showing the effect of Compound 1 on mechanical pain threshold in an L5-L6 spinal nerve ligation animal model;

in fig. 2 is a graph showing the effect on mechanical pain threshold of administering a single dose of Compound 1 to a chronic constriction injury (CCI) model rat (X±s, n=10);

in fig. 3 is a graph showing the effect on mechanical pain threshold of administering a single dose of pregabalin to a rat model of chronic constriction injury (CCI) (X±s, n is 10);

in fig. 4 is a graph showing XRD (X-ray diffraction) of compound 1;

in fig. 5 is a graph showing TGA/DSC of compound 1;

in fig. Figure 6 is a diagram showing the diffraction spectrum of a single crystal of compound 1.

Detailed Description of the Invention

The technical solution of the invention is described in detail below, together with the following graphics and examples, which, however, is only included in the scope of protection of the present invention without limitation.

The structure of the compound was determined using nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). The NMR shift (δ) is given in ppm. (10 ^-6 ). NMR was determined using a nuclear magnetic resonance spectrometer (Bruker Avance III 400 and Bruker Avance 300) with deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ) or deuterated methanol (CD3OD) as solvent and tetramethylsilane (TMS) as internal standard.

MS measurements were performed using an Agilent 6120V (Electrospray Ionization) and Agilent 6120V (Atmospheric Pressure Chemical Ionization).

HPLC (High Performance Liquid Chromatography) measurements were performed using an Agilent 1260 DAD (Diode Array Detector) High Pressure Liquid Chromatograph (ZorbaxSB-C 18 100 x 4.6 mm column).

Known starting materials in the present invention can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as Titan Technologies, Anaji Chemistry, Shanghai Demer, Chengdu Kelong Chemical, Shaoyuan Chemical Technology, Bailingwei Technology and so on.

Microcrystalline cellulose is microcrystalline cellulose 102 unless otherwise noted.

- 11 045716

Intermediate 1: Getting intermediate 1j

Step 1: (±)-(1S,5R,7S)-7-(2-bromoethyl)bicyclo[3.2.0]hept-2-en-6-one (1b)

The starting material 1a (24 g; 0.36 mol) and 1100 ml of cyclohexane were added to the reaction flask under a protective nitrogen atmosphere, and triethylamine (25 g; 0.25 mol) was added thereto and heated to reflux. A solution of 4-bromobutanoyl chloride (46 g; 0.25 mol) in cyclohexane was added dropwise using a syringe pump (100 ml; 25 ml/h), followed by reflux reaction for 4 hours. The reaction solution was filtered by aspiration, washed with cyclohexane (150 mlx3), and the filtrates were combined, washed with saturated ammonium chloride (1000 mlx2) and water (1000 mlx2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography on silica gel (petroleum ether/ethyl acetate (v/v), 80:1) to give light yellow oily product 1b (9.6 g; 18% yield). ^1H NMR (400 MHz, CDCl ₃ ) δ 5.97-5.85 (m, 1H), 5.80-5.70 (m, 1H), 3.91-3.79 (m, 1H), 3.67 (dd, J=9.7, 5, 5 Hz, 2H), 3.47 (t, J=6.8 Hz, 2H), 2.68 (ddd, J=18.3, 15.2, 3.9 Hz, 1H), 2.47-2.31 (m, 1H) , 2.13 (dq, J=21.0, 6.5 Hz, 1H), 1.93 (ddd, J=21.5, 12.2, 7.1 Hz, 1H).

Step 2: (±)-(1S,5R,7S)-7-(2-bromoethyl)spiro[bicyclo[3.2.0]hept-[2]-ene-6,2'-[1,3]dioxolane] (1s)

Compound 1b (23 g; 0.11 mol), para-toluenesulfonic acid monohydrate (1.0 g; 5.5 mmol) and ethylene glycol (27.3 g; 0.44 mol) were placed in a one-neck flask, 250 ml of toluene was added and heated to reflux and separate the water for 6 hours. After cooling, the reaction solution was poured into ice water, sodium bicarbonate was added to adjust the pH to neutral, and extracted with ethyl acetate (300 ml x 3 ). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography (ethyl acetate:petroleum ether, 1:30) to give yellow oily product 1c (21.2 g; 75% yield). 1H NMR (400 MHz, CDCl ₃ ) δ 5.94-5.83 (m, 1H), 5.67-5.56 (m, 1H), 3.95-3.75 (m, 4H), 3.36-3.25 (m, 2H), 3.233.12 ( m, 1H), 3.02 (ddd, J=22.9, 15.7, 8.0 Hz, 2H), 2.48-2.25 (m, 2H), 1.99-1.78 (m, 2H).

Step 3: (±)-(1S,5R,7S)-7-(2-bromoethyl)spiro[bicyclo[3.2.0]hept-[2]-ene-6,2'-[1,3]dioxolαn] -2-ol (1d)

Starting material 1c (15 g; 0.06 mol) was added to the reaction flask, tetrahydrofuran (250 ml) was added as a solvent, and dimethyl borane solution (30 ml; 0.3 mol) was added dropwise in an ice water bath. The resulting mixture was then left to stand at this temperature for 2 hours, purified water (0.6 mol) was added dropwise in an ice water bath, then

- 12 045716 an aqueous solution of sodium hydroxide (3 mol; 1200 ml) was added dropwise, then hydrogen peroxide (containing 0.6 mol H ₂ O ₂ ) was added dropwise. The mixture was then warmed to room temperature to react for 3 hours. Thereafter, it was extracted with ethyl acetate (200 ml x 3) and the organic phases were combined, washed with water (300 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a pale -yellow oily product 1d (16.5 g), which was used in the next step without purification.

Step 4: (±)-(1S,5R,7S)-7-(2-bromoethyl)spiro[bicyclo[3.2.0]hept-[2]-ene-6,2'-[1,3]dioxolane] -2-on (1e)

Compound 1d (16.5 g; 0.06 mol) and dichloromethane (250 ml) were added to the reaction flask, and Dess-Martin reagent was added as an oxidizing agent (38.2 g; 0.09 mol) in one portion in an ice bath, and reacted at room temperature for 2 hours. A saturated sodium bicarbonate solution was added dropwise to the reaction solution until the pH value was approximately 7. The resulting mixture was subjected to liquid separation and the aqueous phase was extracted with dichloromethane (200 mlx2), while the organic phases were combined, washed water (500 mlx1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v), 8:1) to give a light yellow oily product 1e (9.7 g; yield 59%). 1H NMR (400 MHz, CDCl ₃ ) δ 4.02-3.81 (m, 4H), 3.40 (dd, J=10.3, 3.8 Hz, 2H), 3.15 (td, J=10.3, 4.9 Hz, 2H), 2.61 (ddd, J=20.6, 14.0, 8.1 Hz, 2H), 2.27 (ddt, J=18.9, 9.6, 1.8 Hz, 1H), 2.12 -2.00 (m, 1H), 1.99-1.70 (m, 3H).

Step 5: (±)-(1'R,3'S,6'S)-spiro[[1,3]dioxolane-2,2'-tricyclo[4.2.1.0 ^3,8 ]nonane]-7'-one (1f)

H <\T° (+/-) Potassium tert-Butoxide (16 g; 0.14 mol) and tetrahydrofuran (1 L) were added to the reaction flask under a protective nitrogen atmosphere. The resulting mixture was cooled to -0°C, compound 1e in toluene (29 g, 0.11 mol) was added dropwise and then stirred at room temperature for 2 hours. A saturated ammonium chloride solution was added dropwise in an ice bath until the pH was approximately 7, extracted with ethyl acetate (500 mlx2), washed with water (1000 mlx2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v), 10: 1) to give pale yellow oily product 1f (9.5 g; 45% yield). 1H NMR (400 MHz, CDCl3) δ 4.04-3.86 (m, 4H), 3.20-3.07 (m, 1H), 2.99-2.86 (m, 1H), 2.53 (ddd, J=8.6, 5.6, 1.7 Hz, 1H), 2.41-2.24 (m, 2H), 2.24-2.01 (m, 2H), 1.95 (d, J=13.2 Hz, 1H), 1.61 (dddd, J=14.4, 7.6, 2.6, 0.7 Hz, 1H), 1.51-1.38 (m, 1H).

Step 6: (±)-(1'R,3'S,6'S)-spiro[[1,3]dioxolane-2,2'-tricyclo[4.2.1.0 ^3,8 ]nonane] (1g)

H A-A-o (+/-)

Starting material 1f (9.0 g; 46.3 mmol) and diethylene glycol (150 mL) were added to the reaction flask, and hydrazine hydrate (8.9 g; 278 mmol) and potassium hydroxide (15.6 g; 278 mmol) were added. The resulting mixture was reacted at 180°C for 3 hours, followed by rotary evaporation under reduced pressure to remove water at 70°C, then heated to 220°C, and reacted for 2 hours, and cooled. Water (200 ml) was added to the resulting reaction solution, extracted with methyl tert-butyl ether (300 mlx3), washed with 1 mol/l hydrochloric acid (400 mlx2), washed with water (400 mlx2), dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v), 60:1) to give a colorless oily product 1g (3 g), which was used in the next step without purification.

- 13 045716

Step 7: (±)-(1R,3S,6R,8R)-tricyclo[4.2.1.0 ^3,8 ]nonan-2-one (1h)

Starting material 1g (3 g; 16.6 mmol) was added to the reaction flask, then tetrahydrofuran (36 ml) and water (12 ml) were added as a solvent, and trifluoroacetic acid (8 ml) was added dropwise in an ice bath, and subjected to reaction at 45°C for 3 hours. The resulting mixture was added dropwise with a saturated solution of sodium bicarbonate in an ice bath until the pH value was approximately 7, then extracted with ethyl acetate (80 mlx3), washed with water (100 mlx2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v), 100:1) to give a white solid 1h (2 g; 88% yield). ^1H NMR (400 MHz, CDCl ₃ ) δ 3.47-3.33 (m, 1H), 3.19 (dd, J=3.3, 1.8 Hz, 1H), 2.84-2.69 (m, 1H), 2.47-2.32 (m, 1H), 2.12-1.97 (m, 1H), 1.93 (d J=12.3 Hz, 1H), 1.82-1.69 (m, 1H), 1.56-1.35 (m, 4H), 1.27-1.10 ( m, 1H).

Step 8: (±)-ethyl-2-((1R,3S,6R,8R)-tricyclo[4.2.1.0 ^3,8 ]non-2-ylidene)acetate (1i)

Sodium hydride (60%; 91.6 g; 3.82 mol) and tetrahydrofuran (5 L) were added to the reaction flask, cooled to 0°C and ethyl diethoxyphosphonoethyl acetate (856 g; 3.82 mol) in tetrahydrofuran (400 ml). The resulting mixture was then left to stand at this temperature for 15 minutes and compound 1h (400 g, 2.94 mol) in tetrahydrofuran (200 ml) was added dropwise and then warmed to room temperature to react for 1 hour. The resulting mixture was added dropwise with saturated ammonium chloride to pH 7-8 in an ice water bath, extracted with ethyl acetate (500 ml x 3), washed with saturated brine (500 ml x 2), dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography on silica gel (petroleum ether/ethyl acetate (v/v), 30:1) to give light yellow oily product 1i (310 g; 51% yield).

Step 9: (±)-ethyl-2-((1R,3S,6R,8R)-2-(nitromethyl)tricyclo[4.2.1.0 ^3,8 ]-non-2-yl)acetate (1i)

Starting material 1i (390 g, 1.89 mol), nitromethane (4 L) and 1,8-diazabicyclo-[5.4.0]undec-7-ene (575.6 g, 3.78 mol) were added to the reaction flask and heated to 80°C for 9 hours. The reaction mixture was poured into ice water (3000 ml), extracted with DCM (2000 mlx3), washed with brine (3000 ml), dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether/ethyl acetate (vol). /vol.), 100:1) to obtain a colorless oily product 1j (360 g; yield 71%).

Receiving connection 1

Chiral separation

Connection 1

11-1 (±)-Ethyl-2-((1R,3S,6R,8R)-2-(nitromethyl)tricyclo[4.2.1.0 ^3,8 ] non-2-yl)acetate (intermediate 1j) (360 d) used for separation under the following conditions: Thar analytical SFC (SFC-A) instrument, chiralPak AD column, 150x4.6 mm ID (inner diameter), 3 µm, with mobile phases A for CO ₂ and B for methanol, gradient B 5-40%, flow rate 2.4 ml/min and column temperature 35°C. After separation, two optical isomers were obtained: peak 1 (retention time: 3.8 min, 174 g) and peak 2

- 14 045716 (retention time: 5.7 min, 160 g). Compound 1j: [α] 20D=0.00° (C=0.9, CH ₂ Cl ₂ ); Peak 2: [α] 20D=44° (C=0.103, CH ₃ OH), where C refers to the mass (in grams) of the test substance per 100 ml of solution, and 20D refers to the test at 20°C, with a sodium lamp in as a light source, at a wavelength of 589 nm. Step 1: (1^,2^,3'K6^,8'8)-spiro[pyrrolidine-3,2-tricyclo[4.2.1.0 ^3,8 ]-nonane]-5-one (1k-1)

Starting material 1j-1 (peak 2; 270 g; 1.01 mol), ethanol (1 L) and water (1 L) were added to the reaction flask, then reduced iron powder (282 g; 5.05 mol) and chloride were added ammonium (162 g; 3.03 mol) and reacted at reflux for 4 hours. The reaction solution was filtered and the filtrate was concentrated to remove ethanol, after which 500 ml of water was added to the remaining solution while the filter cake was washed with dichloromethane (200 ml x 3 ). The filtrate was collected and the organic phase was mixed with the above remaining solution, subjected to liquid separation, extracted twice with dichloromethane (500 mlx2). The organic phases were combined, washed with water (500 mlx2), dried with sodium sulfate, concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane/methanol (v/v), 40:110:1) to give a white solid 1k -1 (160 g; yield 83%).

Step 2: 2-((1S,2S,3R,6S,8S)-2-(aminomethyl)tricyclo[4.2.1.0 ^3,8 ]non-2-yl)-acetic acid (1l-1)

Starting material 1k-1 (320 g, 1.673 mol) was added to the reaction flask, then 6N hydrochloric acid (1.6 L) was added and refluxed for 16 hours. The precipitated solid was filtered off. The resulting solid was dissolved in 1 L of purified water, the pH was adjusted to approximately 7 with concentrated aqueous ammonia, aspiration filtered, washed with ice water and dried to obtain a white solid. The pH of the filtrate was adjusted to approximately 6 with 10 N sodium hydroxide in an ice water bath, and further adjusted to approximately 7 with concentrated aqueous ammonia, and extracted with dichloromethane (1 Lx3). The remaining aqueous phase was concentrated and dried, filtered and washed with ice water to obtain a white solid. The solid obtained in two portions was triturated with dichloromethane (1.5 Lx3) to give a white solid 11-1 (245 g; 70%).

Receiving connection 1

Compound 1l-1 (245 g; 1.17 mol) was added to the reaction flask, methanol (2.2 L) was added, and benzenesulfonic acid monohydrate (268.0 g; 1.52 mol) in methanol was added dropwise, then stirred for 1 hour at room temperature. The precipitated solid was filtered off by aspiration, and the filtrate was concentrated to obtain a solid. The solid obtained in two portions was combined and triturated with dichloromethane (1.5 Lx3), filtered, washed with ethyl acetate and dried to give pure compound 1 (398 g; yield 92.5%, HPLC: 99%, chemical formula C ₁₂ H ₁₉ NO ₂ -C ₆ H ₆ O ₃ S).

^1H NMR (400 MHz, D2O) δ 7.85-7.70 (m, 2H), 7.54 (tt, J=14.3, 7.2 Hz, 3H), 3.33 (d, J=13.8 Hz, 2H) , 2.81 (dd, J=13.2, 5.4 Hz, 1H), 2.57 (q, J=17.6 Hz, 2H), 2.47-2.37 (m, 1H), 2.27 (dd, J=12, 0, 6.0 Hz, 1H), 2.17-2.06 (m, 1H), 1.96 (dd, J=21.6, 9.5 Hz, 1H), 1.79-1.66 (m, 1H), 1.66-1.40 ( m, 4H), 1.33 (dd, J=14.3, 9.0 Hz, 1H), 1.26-1.15 (m, 1H).

The peak magnitude of Compound 1 in the X-ray powder diffraction (XRD) pattern is shown in the table below and specifically shown in FIG. 4.

- 15 045716

20 Peak height FWHM (full line width at half height) Interplanar distance [A] Relative peak height [%] 9.7167 4046.64 0.1279 9.10278 33.05 11.1352 1612.11 0.0384 7.94615 13.17 11.2128 1820.68 0.1023 7.89135 14.87 11.8322 277.75 0.1535 7.47961 2.27 14.0077 3132.13 0.0512 6.32246 25.58 14.0962 2255.84 0.064 6.28296 18.43 15.1565 3670.71 0.064 5.84573 29.98 15.3713 1403.94 0.0768 5.76452 11.47 15.6037 1165.35 0.1279 5.67918 9.52 15.9607 443.05 0.0512 5.55295 3.62 16.3325 12242.87 0.1023 5.42737 100 18.2092 900.66 0.1151 4.87203 7.36 18.87 1903.14 0.064 4.70287 15.54 19.0176 2554.83 0.0384 4.66672 20.87 19.3165 5667.72 0.1279 4.59518 46.29 19.8771 4571.39 0.0895 4.46681 37.34 20.3279 1930.15 0.0384 4.36878 15.77 20.4622 3513.85 0.0768 4.3404 28.7 21.1053 859.6 0.0768 4.20958 7.02 21.2599 1641.19 0.0512 4.17932 13.41 21.5253 3785.86 0.0384 4.12837 30.92 21.6885 7208.47 0.1151 4.09769 58.88 22.3935 724.03 0.1535 3.97025 5.91 22.7865 262.87 0.0384 3.90265 2.15 23.4692 1810.44 0.0895 3.79065 14.79 24.1631 54.8 0.1535 3.68334 0.45 25.3274 1659.16 0.0624 3.51369 13.55 25.4027 1478.13 0.0512 3.50635 12.07 25.6773 1433.37 0.0512 3.46946 11.71 25.9344 1421.23 0.0468 3.4328 11.61 25,999 1046.47 0.0468 3.43294 8.55 26.9314 950.92 0.1404 3.30794 7.77 27.3414 296.52 0.1248 3.25927 2.42 27.9554 2439.22 0.0468 3.18906 19.92 28.0286 2418.82 0.0624 3.1809 19.76 28.5766 1038.14 0.078 3.12113 8.48 29.1738 996.82 0.0468 3.05858 8.14 29.8174 1938.81 0.1248 2.99402 15.84 30.6914 348.82 0.0468 2.91071 2.85 31.3063 653.82 0.1248 2.85493 5.34 31.5601 212.27 0.0936 2.83255 1.73 32.3291 96.39 0.1872 2.76691 0.79 33.0473 214.64 0.1872 2.7084 1.75 34.2421 109.66 0.1248 2.61658 0.9 35,264 437.3 0.078 2.54306 3.57 35.5326 412.09 0.0468 2.52445 3.37 36.1622 418.43 0.156 2.48193 3.42 36.6015 376.67 0.1872 2.45315 3.08 37,239 295.68 0.156 2.4126 2.42 37,714 195.04 0.1092 2.3833 1.59 35.5326 412.09 0.0468 2.52445 3.37

The TGA/DSC spectra of compound 1 are shown in FIG. 5, and the diffraction spectrum of a single crystal of compound 1 is shown in FIG. 6.

Example 1. Compatibility of excipient and compound 1.

Mixtures of the active substance, compound 1, with various types of fillers were prepared according to table. 1 and kept open under conditions of 75% relative humidity and a temperature of 40°C,

- 16 045716 then measured the content of impurities in them, respectively. The results of the study on the content of impurities are shown in table. 2.

Table 1

Mixtures of compound 1 with various types of fillers

Substances Scheme 1 Scheme 2 Scheme 3 Scheme 4 Scheme 5 Scheme 6 Scheme 7 Scheme 8 Connection 1 1 part 1 part 1 part 1 part 1 part 1 part 1 part 1 part Mannitol 5 parts 0 0 0 0 0 0 0 Lactose 0 5 parts 0 0 0 0 0 0 Starch 0 0 5 parts 0 0 0 0 0 Pregelatinized starch 0 0 0 5 parts 0 0 0 0 Sorbitol 0 0 0 0 5 parts 0 0 0 Sucrose 0 0 0 0 0 5 parts 0 0 Hydroxypropyl cellulose with low degree of substitution 0 0 0 0 0 0 5 parts 0 Microcrystalline cellulose 0 0 0 0 0 0 0 5 parts

The preparation of a mixture of compound 1 with various types of fillers was carried out as follows:

1) passing Compound 1 through a 60 mesh sieve and passing the filler through a 40 mesh sieve for use;

2) uniformly mixing compound 1 and filler to obtain a mixed powder;

3) keeping the sample open under conditions of 75% relative humidity and a temperature of 40°C.

table 2

Change in the content of impurities in a mixture of compound 1 with various fillers

No. Total impurity content, % 0 day 7 days 14 days 30 days Scheme 1 0.054 0.056 0.064 0.173 Scheme 2 0.061 0.068 0.166 0.297 Scheme 3 0.054 0.068 0.123 0.282 Scheme 4 0.059 0.139 0.272 0.381 Scheme 5 0.064 0.078 0.148 0.298 Scheme 6 0.062 0.185 0.312 0.425 Scheme 7 Scheme 8 0.068 0.098 0.112 0.188 0.042 0.056 0.057 0.063

Conclusion: Mannitol, low-substituted hydroxypropylcellulose, and microcrystalline cellulose have good compatibility with compound 1.

Example 2. The influence of various types of fillers on the homogeneity of mixing the drug in the capsule.

Capsules with compound 1 are obtained in accordance with table. 3 through the following manufacturing method:

1) passing Compound 1 through a 60 mesh sieve and passing the filler through a 40 mesh sieve for use;

2) adding sifted compound 1 and filler to the mixer with multidirectional movement for uniform mixing;

3) adding binder, disintegrant and glidant to the multi-directional mixer to mix evenly with the mixture from step (2);

- 17 045716

4) adding lubricant to the multi-directional mixer for final mixing with the mixture from step (3);

5) selecting the appropriate type of capsule and filling the capsule with the finished mixture obtained in step (4).

Table 3

The drug is in the form of capsules with various fillers

No. Starting materials Quantity Composition of a single dose, in mass percent Functions Drug 1 Compound 1 (in terms of Ci2Hi9NO2-C6H ₆ O3S) 1 part 4.08 Active substance Mannitol 20 parts 81.63 Filler Hydroxypropyl cellulose 1 part 4.08 Binder Silica 1 part 4.08 Sliding agent Croscarmellose sodium 1 part 4.08 Baking powder Magnesium stearate 0.5 parts 2.04 Lubricant Gelatin capsule 1 capsule 1 capsule — Drug 2 Compound 1 (in terms of C1 ₂ H1 ₉ M)2-SbNbOz8) 1 part 4.08 Active substance Hydroxypropyl cellulose with low degree of substitution 20 parts 81.63 Filler Hydroxypropyl cellulose 1 part 4.08 Binder Silica 1 part 4.08 Sliding agent Croscarmellose sodium 1 part 4.08 Baking powder Magnesium stearate 0.5 parts 2.04 Lubricant Gelatin capsule 1 capsule 1 capsule — Preparation 3 Compound 1 (in terms of Ci2Hi9NO2-C ₆ H ₆ O3S) 1 part 4.08 Active substance Microcrystalline cellulose 12 20 parts 81.63 Filler Hydroxypropyl cellulose 1 part 4.08 Binder substance Silica 1 part 4.08 Sliding agent Croscarmellose sodium 1 part 4.08 Baking powder Magnesium stearate 0.5 parts 2.04 Lubricant Gelatin capsule 1 capsule 1 capsule —

Compound 1 capsules were analyzed by high-performance liquid chromatography according to General Article 0941 Test Method for Dosage Uniformity of the Chinese Pharmacopoeia, 2015 edition. The results are shown in table. 4.

- 18 045716

Table 4 with connection 1

No. Value A+2.2S RSD (relative standard deviation), % Drug 1 10.75 6.56 Drug 2 12.64 7.42 Preparation 3 8.42 4.38

Example 3. The effect of a combination of different types of excipients on the stability of the drug and the uniformity of dosing.

Capsules with compound 1 were obtained in accordance with the proportions given in table. 5, through the manufacturing method as shown in Example 2, and their dosage uniformity and stability were measured.

Table 5

Starting materials for capsules with compound 1

Composition of a single No. Starting materials Quantity doses, in mass Functions percent Compound 1 (in terms of Active Preparation 4 1 part 3.92 substance on C12N1 ₉ M)2-SbNbOz8) Mannitol 7 parts 27.45 Filler Hydroxypropyl cellulose with 7 parts 27.45 Filler low degree of substitution Microcrystalline 7 parts 27.45 Filler cellulose 102 — Binder Hydroxypropyl cellulose 1 part 3.92 substance Sliding Silica 1 part 3.92 agent Croscarmellose sodium 1 part 3.92 Baking powder Lubricating Magnesium stearate 0.5 parts 1.96 substance Gelatin capsule 1 capsule 1 capsule — Compound 1 (in terms of 1 part 3.92 Active to CnHwNCh'CeHeChS) substance Mannitol 0 parts 0.00 Filler Hydroxypropyl cellulose with low degree of substitution 7 parts 27.45 Filler Microcrystalline 14 parts 54.90 Filler Preparation 5 cellulose 102 — Binder Hydroxypropyl cellulose 1 part 3.92 substance Sliding Silica 1 part 3.92 agent Croscarmellose sodium 1 part 3.92 Baking powder Lubricating Magnesium stearate 0.5 parts 1.96 substance Gelatin capsule 1 capsule 1 capsule — Compound 1 (in terms of 1 part 3.92 Active to CnHwNCh'CeHeChS) substance Preparation 6 Mannitol 7 parts 27.45 Filler Hydroxypropyl cellulose with low degree of substitution 0 parts 0.00 Filler Microcrystalline 14 parts 54.90 Filler

- 19 045716

cellulose 102 Hydroxypropyl cellulose 1 part 3.92 Binder Silica 1 part 3.92 Sliding agent Croscarmellose sodium 1 part 3.92 Baking powder Magnesium stearate 0.5 parts 1.96 Lubricant Gelatin capsule 1 capsule 1 capsule — Compound 1 (in terms of Ci2Hi9NO2-C6H ₆ O3S) 1 part 3.92 Active substance Mannitol 7 parts 27.45 Filler Hydroxypropyl cellulose with low degree of substitution 14 parts 54.90 Filler Microcrystalline cellulose 102 0 parts 0.00 Filler Preparation 7 Hydroxypropyl cellulose 1 part 3.92 Binder Silica 1 part 3.92 Sliding agent Croscarmellose sodium 1 part 3.92 Baking powder Magnesium stearate 0.5 parts 1.96 Lubricant Gelatin capsule 1 capsule 1 capsule — Compound 1 (in terms of Ci2Hi9NO2-C6H ₆ O3S) 1 part 9.52 Active substance Mannitol 3 parts 28.57 Filler Preparation 8 Microcrystalline cellulose 102 3 parts 28.57 Filler Hydroxypropyl cellulose 1 part 9.52 Binder Silica 1 part 9.52 Sliding agent Croscarmellose sodium 1 part 9.52 Baking powder Magnesium stearate 0.5 parts 4.76 Lubricant Gelatin capsule 1 capsule 1 capsule — Compound 1 (in terms of C12H19NO2-C6H6O3S) 1 part 1.55 Active substance Mannitol 30 parts 46.51 Filler Microcrystalline cellulose 102 30 parts 46.51 Filler Preparation 9 Hydroxypropyl cellulose 1 part 1.55 Binder Silica 1 part 1.55 Sliding agent Croscarmellose sodium 1 part 1.55 Baking powder Magnesium stearate 0.5 parts 0.78 Lubricant Gelatin capsule 1 capsule 1 capsule —

Compound 1 capsules obtained in Example 3 were analyzed by high-performance liquid chromatography according to General Article 0941 Dosage Uniformity Test Method of the Chinese Pharmacopoeia, 2015 edition. The results are shown in Table. 6.

- 20 045716

Table 6

Uniformity of dosing of capsules with compound 1_____

No. Value A+2.2S RSD, % Preparation 4 6.25 2.53 Preparation 5 6.34 2.18 Preparation 6 4,I 1.52 Preparation 7 5.48 2.45 Preparation 8 10.25 4.13 Preparation 9 12.34 4.28

Determination of impurities was carried out by keeping the samples open in conditions of 75% relative humidity and a temperature of 40°C to measure the content of impurities in them. The results of the determination are shown in table. 7.

Table 7

Stability of capsules with compound 1

No. Total impurity content, % 0 day 7 days 14 days 30 days Preparation 4 0.054 0.153 0.284 0.573 Preparation 5 0.061 0.098 0.256 0.547 Preparation 6 0.057 0.061 0.201 0.372 Preparation 7 0.059 0.139 0.372 0.781

The results show that impurity levels in formulations 4-7 after 30 days were high.

Example 4. Wet granulation in the manufacture of capsules with compound 1.

Capsules with compound 1 were obtained in accordance with the compositions given in table. 8, through the following preparation method:

1) passing Compound 1 through a 60 mesh sieve and passing mannitol and microcrystalline cellulose through a 40 mesh sieve for use;

2) preparation of hydroxypropylcellulose in a 40% aqueous solution of ethanol as a binder;

3) adding compound 1, mannitol and microcrystalline cellulose to a wet granulator, and adding the binder obtained in step (2) thereto for shear mixing to obtain a soft material;

4) passing the soft material through a sieve to obtain particles and then subjecting it to static drying;

5) granulating the dried soft material and then adding it with croscarmellose sodium and silica into a multi-directional mixer for uniform mixing;

6) adding magnesium stearate to the multi-directional mixer for final mixing; And

7) selecting the appropriate capsule type and filling the capsule with the mixed powder.

Table 8

Starting materials for capsules with compound 1_______

No. Starting materials Quantity Mass percentage(%) Functions Preparation 10 Compound 1 (in terms of CnHwNCh'CeHeChS) 1 part 3.92 Active substance Mannitol 7 parts 27.45 Filler Microcrystalline cellulose 102 14 parts 54.90 Filler Hydroxypropyl cellulose 1 part 3.92 Binder Silica 1 part 3.92 Sliding agent Croscarmellose sodium 1 part 3.92 Baking powder Magnesium stearate 0.5 parts 1.96 Lubricant Gelatin capsule 1 capsule 1 capsule —

- 21 045716

The resulting Compound 1 capsules were analyzed by high-performance liquid chromatography according to General Article 0941 Test Method for Dosage Uniformity of the Chinese Pharmacopoeia, 2015 Edition. The results are shown in table. 9.

Table 9

Uniformity of dosing of capsules with compound 1

No. Value A+2.2S RSD, % Preparation 10 5.42 2.54

Conclusion: The resulting composition has an A+2.2S value less than or equal to 15, which meets the requirements described in the Chinese Pharmacopoeia, 2015 edition, for dosage uniformity.

The resulting Compound 1 capsules were subjected to a dissolution test using high performance liquid chromatography, and the results are shown in Table. 10.

Table 10

Dissolution of the capsule with compound 1

No. Dissolution rate after 15 minutes (%) Preparation 10 92.5

Conclusion: the resulting composition can be quickly dissolved.

Example 5. Wet granulation in the manufacture of capsules with compound 1.

Capsules with compound 1 were obtained in accordance with the compositions given in table. 11, through the following manufacturing method:

1) passing Compound 1 through a 60 mesh sieve and passing mannitol and microcrystalline cellulose through a 40 mesh sieve for use;

2) preparing a 40% aqueous solution of ethanol as a solvent;

3) adding compound 1, hydroxypropyl methylcellulose, mannitol and microcrystalline cellulose to a wet granulator, and adding the solvent obtained in step (2) thereto for shear mixing to obtain a soft material;

4) passing the soft material through a sieve to obtain particles, and then performing dynamic drying in a fluidized bed;

5) granulating the dried soft material and then adding it with croscarmellose sodium and silica into a multi-directional mixer for uniform mixing;

6) adding magnesium stearate to the multi-directional mixer for final mixing;

7) selecting the appropriate capsule type and filling the capsule with the mixed powder.

Table 11

Starting materials for capsules with compound 1

No. Starting materials Quantity Mass percentage(%) Functions Preparation 11 Compound 1 (in terms of 1 part 3.92 Active on Ci2Hi9NO2-C6H ₆ O3S) substance Mannitol 7 parts 27.45 Filler Microcrystalline cellulose 102 14 parts 54.90 Filler Hydroxypropyl cellulose 1 part 3.92 Binder Silica 1 part 3.92 Sliding agent Croscarmellose sodium 1 part 3.92 Baking powder Magnesium stearate 0.5 parts 1.96 Lubricant Gelatin capsule 1 capsule 1 capsule —

The resulting Compound 1 capsules were analyzed by high-performance liquid chromatography according to General Article 0941 Test Method for Dosage Uniformity of the Chinese Pharmacopoeia, 2015 Edition. The results are shown in Table. 12.

- 22 045716

Table 12

Uniformity of dosing of capsules with compound 1

No. Value A+2.2S RSD, % Preparation 11 6.41 1.78

Conclusion: The resulting composition has an A+2.2S value less than or equal to 15, which meets the requirements described in the Chinese Pharmacopoeia, 2015 edition, for dosage uniformity.

The resulting Compound 1 capsules were subjected to a dissolution test using high performance liquid chromatography, and the results are shown in Table. 13.

Table 13 _______Dissolution of capsule with compound 1_______

No. Dissolution rate after 15 minutes (%) Preparation 11 92.4

Conclusion: the resulting composition can be quickly dissolved. Capsules with compound 1 were obtained in accordance with table. 14 by the method described in Example 5. The resulting capsules had dosage uniformity meeting the requirements described in the Chinese Pharmacopoeia, 2015 edition.

Table 14

Starting materials and capsule compositions with compound 1

No. Active substance Filler 1 Filler 2 Binder Sliding agent Baking powder Lubricant Capsule shell A drug 12 1 part Mannitol 30 parts MSS 30 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 13 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 14 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part CMCS 1 part Magnesium stearate 0.5 parts One gelatin capsule A drug 15 1 part Mannitol 7 parts MSS 14 parts LDC 0.1 parts Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 16 1 part Mannitol 7 parts MSS 14 parts LDC 3 parts Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 17 1 part Mannitol 7 parts MSS 14 parts HPMC 0.1 parts Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 18 1 part Mannitol 7 parts MSS 14 parts HPMS 3 parts Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 19 1 part Mannitol 7 parts MSS 14 parts Povidone 0.1 part Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 20 1 part Mannitol 7 parts MSS 14 parts Povidone 5 parts Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 21 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 5 parts SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 22 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Talc 1 part SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 23 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Talc 5 parts SMS part 1 Magnesium stearate 0.5 parts One gelatin capsule A drug 24 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Magnesium stearate 2 parts One gelatin capsule A drug 25 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.1 part One gelatin capsule A drug 26 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Stearic acid 2 parts One gelatin capsule A drug 27 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Stearic acid 0.1 part One gelatin capsule A drug 1 part Mannitol MSS LDC Dioxide SMS SSF One

- 23 045716

28 7 parts 14 parts 1 part silicon 1 part 1 part 2 parts gelatin capsule A drug 29 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 SSF 0.1 parts One gelatin capsule A drug thirty 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Glycerin behenate 2 parts One gelatin capsule A drug 31 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Glycerol behenate 0.1 parts One gelatin capsule A drug 32 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One empty HPS capsule A drug 33 1 part Mannitol 7 parts MSS 14 parts LDC part 1 Silicon dioxide 1 part SMS part 1 Magnesium stearate 0.5 parts One HPMC capsule

Abbreviations in the table above: MCC: microcrystalline cellulose; HPC: hydroxypropylcellulose; HPMC: hydroxypropyl methylcellulose; CMS: croscarmellose sodium; HPS: hydroxypropyl starch; SSF: sodium stearyl fumarate.

Example 6. Capsules with compound 1.

Capsules with compound 1 were obtained in accordance with the compositions given in table. 15, through the following manufacturing method:

1) passing the active pharmaceutical ingredient, compound 1, through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use;

2) adding compound 1 and filler to the mixer with multidirectional movement for uniform mixing;

3) adding lubricant and other auxiliary substances to the multi-directional mixer for final mixing with the mixture obtained in step (2);

4) filling the capsule with mixed powder.

Table 15

Starting materials and capsule compositions with compound 1

No. Starting materials Quantity Mass percentage(%) Functions Preparation 34 Compound 1 (in terms of C12N19ХО2-SbN ₆ Oz8) 1 part 5.52 — Mannitol 17 parts 93.92 Filler Magnesium stearate 0.1 parts 0.55 Lubricant Empty gelatin capsule 1 capsule 1 capsule — Drug 3 5 Compound 1 (in terms of SpNshchhS-SbNbOzZ) 1 part 5.52 — Microcrystalline cellulose 102 17 parts 93.92 Filler Magnesium stearate 0.1 parts 0.55 Lubricant Empty gelatin capsule 1 capsule 1 capsule — Preparation 36 Compound 1 (in terms of C12H1 ₉ SiO2'SbHbOz8) 1 part 6.06 — Mannitol 5 parts 30.30 Filler Microcrystalline cellulose 102 10 parts 62.11 Filler Magnesium stearate 0.1 parts 0.62 Lubricant Empty gelatin capsule 1 capsule 1 capsule —

The resulting Compound 1 capsules were analyzed by high-performance liquid chromatography according to General Article 0941 Test Method for Dosage Uniformity of the Chinese Pharmacopoeia, 2015 Edition. The results are shown in table. 16.

- 24 045716

Table 16

Uniformity of dosing of capsules with compound 1

No. Value A+2.2S capsules RSD of mixed powders, % Preparation 34 18.9 9.64 Preparation 35 21.4 12.5 Preparation 36 15.4 6.24

Conclusion: Drugs 34, 35 and 36 have an A+2.2S value greater than or equal to 15, which does not meet the requirements described in the Chinese Pharmacopoeia, 2015 edition for dosage uniformity.

Example 7. Capsules with compound 1.

Capsules with compound 1 were obtained in accordance with the compositions given in table. 17, through the following preparation method:

1) passing the active pharmaceutical ingredient, compound 1, through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use;

2) adding compound 1 and filler to the mixer with multidirectional movement for uniform mixing;

3) adding lubricant and other auxiliary substances to the multi-directional mixer for final mixing with the mixture obtained in step (2);

4) filling the capsule with mixed powder.

Table 17

Starting materials and capsule compositions with compound 1

No. Starting materials Quantity Mass percentage(%) Functions A drug 37 Compound 1 (in terms of Ci2Hi9NO2-C6H ₆ O ₃ S) 1 part 5.85 — Mannitol 5 parts 29.24 Filler Microcrystalline cellulose 102 10 parts 58.48 Filler Hydroxypropyl cellulose 1 part 5.85 Binder Magnesium stearate 0.1 parts 0.58 Lubricant Empty gelatin capsule 1 capsule 1 capsule — A drug 38 Compound 1 (in terms of Ci2Hi9NO2-C ₆ H ₆ O ₃ S) 1 part 5.85 — Mannitol 5 parts 29.24 Filler Microcrystalline cellulose 102 5 parts 29.24 Filler Microcrystalline cellulose 101 5 parts 29.24 Filler Hydroxypropyl cellulose 1 part 5.85 Binder Magnesium stearate 0.1 parts 0.58 Lubricant Empty gelatin capsule 1 capsule 1 capsule —

Determination of impurities: The sample was kept at 75% relative humidity and 40°C (open) to determine the impurities in it. Changes in the total impurity content are shown in table. 18.

- 25 045716

Table 18

Change in the total content of impurities in capsules with compound 1

Sample No. Total impurity content, % 0 day 7 days 14 days 30 days Preparation 37 0.051 0.065 0.148 0.342 Drug 3 8 0.054 0.066 0.164 0.387

The resulting Compound 1 capsules were analyzed by high-performance liquid chromatography according to General Article 0941 Test Method for Dosage Uniformity of the Chinese Pharmacopoeia, 2015 Edition. The results are shown in table. 19.

Table 19

Uniformity of dosing of capsules with compound 1

No. Value A+2.2S capsules RSD of mixed powders, % Preparation 37 10.5 5.44 Preparation 38 8.42 4.29

From the above results it can be seen that the compatibility of the active substance and the excipients was improved. Example 8. Capsules with compound 1.

Capsules with compound 1 were obtained in accordance with the compositions given in table. 20, through the following manufacturing method:

1) passing the active pharmaceutical ingredient, compound 1, through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use;

2) adding compound 1 and filler to the mixer with multidirectional movement for uniform mixing;

3) adding lubricant and other auxiliary substances to the multi-directional mixer for final mixing with the mixture obtained in step (2);

4) filling the capsule with mixed powder.

Table 20

Starting materials and capsule compositions with compound 1

No. Starting materials Quantity Mass percentage(%) Functions A drug 39 Compound 1 (in terms of Ci2Hi9NO2-C6H ₆ O3S) 1 part 6.21 — Mannitol 5 parts 31.06 Filler Microcrystalline cellulose 102 5 parts 31.06 Filler Microcrystalline cellulose 103 5 parts 31.06 Filler Magnesium stearate 0.1 parts 0.62 Lubricant Empty gelatin capsule 1 capsule 1 capsule — A drug 40 Compound 1 (in terms of Ci2Hi9NO2-C6H ₆ O3S) 1 part 6.21 — Mannitol 5 parts 31.06 Filler Microcrystalline cellulose 102 5 parts 31.06 Filler Microcrystalline cellulose 112 5 parts 31.06 Filler Magnesium stearate 0.1 parts 0.62 Lubricant Empty gelatin capsule 1 capsule 1 capsule — A drug 41 Compound 1 (in terms of Ci2Hi9NO2-C ₆ H ₆ O3S) 1 part 6.21 — Mannitol 5 parts 31.06 Filler Microcrystalline 5 parts 31.06 Filler

- 26 045716

cellulose 102 Microcrystalline cellulose 14 5 parts 31.06 Filler Magnesium stearate 0.1 parts 0.62 Lubricant Empty gelatin capsule 1 capsule 1 capsule — A drug 42 Compound 1 (in terms of Ci2Hi9NO2-C6H ₆ O3S) 1 part 6.21 — Mannitol 5 parts 31.06 Filler Microcrystalline cellulose 102 5 parts 31.06 Filler Microcrystalline cellulose 12 5 parts 31.06 Filler Magnesium stearate 0.1 parts 0.62 Lubricant Empty gelatin capsule 1 capsule 1 capsule — A drug 43 Compound 1 (in terms of C12H19NO2-C6H6O3S) 8 parts 44.2 — Microcrystalline cellulose 102 7.6 parts 41.99 Filler Microcrystalline cellulose 12 0.6 parts 3.31 Filler Mannitol 1.8 parts 9.94 Filler Magnesium stearate 0.1 parts 0.55 Lubricant Empty gelatin capsule 1 capsule — A drug 44 Compound 1 (in terms of Ci2Hi9NO2-C ₆ H ₆ O3S) 1 part 5.85 — Microcrystalline cellulose 102 3 parts 17.54 Filler Microcrystalline cellulose 12 10 parts 58.48 Filler Mannitol 3 parts 17.54 Filler Magnesium stearate 0.1 parts 0.58 Lubricant Empty gelatin capsule 1 capsule — A drug Compound 1 (in terms of Ci2Hi9NO2-C ₆ H ₆ O3S) 1 part 5.26 — 45 Microcrystalline cellulose 102 13 parts 68.42 Filler Microcrystalline cellulose 12 1 part 5.26 Filler Mannitol 3 parts 15.79 Filler Magnesium stearate 1 part 5.26 Lubricant Empty gelatin capsule 1 capsule —

The resulting Compound 1 capsules were analyzed by high-performance liquid chromatography according to General Article 0941 Test Method for Dosage Uniformity of the Chinese Pharmacopoeia, 2015 Edition. The results are shown in table. 21.

- 27 045716

Table 21

Uniformity of dosing of capsules with compound 1

No. Value A+2.2S capsules RSD of mixed powders, % Preparation 39 15.3 8.47 Preparation 40 P,4 7.58 Preparation 41 10.7 7.4 Preparation 42 8.5 5.18 Preparation 43 4.67 1.21 Preparation 44 6.27 2.87 Preparation 45 7.24 0.97

Conclusion: Drug 39 has an A+2.2S value greater than 15, which does not meet the requirements described in the Chinese Pharmacopoeia 2015 edition for dosage uniformity. Drugs 42, 43, 44 and 45 have been improved in dosage uniformity.

Example 9. Preparation of drugs in the form of capsules according to the specification for 5 and 20 mg of the active substance (in terms of the free base of compound 1).

Capsules with compound 1 were obtained in accordance with the compositions given in table. 22, through the following manufacturing method:

1) passing the active pharmaceutical ingredient, Compound 1, through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use;

2) adding compound 1 and filler to the mixer with multidirectional movement for uniform mixing;

3) adding lubricant to the multi-directional mixer for final mixing with the mixture obtained in step (2);

4) filling the capsule with mixed powder.

- 28 045716

Table 22

Capsule compositions with compound 1

No. Starting materials Quantity Mass percentage(%) Functions Drug 46 (5 mg) Compound 1 (in terms of CnHwNCh-CeHeChS) 1 part 5.52 Active substance Microcrystalline cellulose 102 13 parts 71.82 Filler Microcrystalline cellulose 12 1 part 5.52 Filler Mannitol 3 parts 16.57 Filler Magnesium stearate 0.1 parts 0.55 Lubricant Empty gelatin capsule 1 capsule 1 capsule — Drug 47 (5 mg) Compound 1 (in terms of CnHwNCh-CeHeChS) 1 part 5.52 Active substance Microcrystalline cellulose 102 11 parts 60.77 Filler Microcrystalline cellulose 12 1 part 5.52 Filler Mannitol 5 parts 27.62 Filler Magnesium stearate 0.1 parts 0.55 Lubricant Empty gelatin capsule 1 capsule 1 capsule — Preparation 48 (20 mg) Compound 1 (in terms of CuHigNCh-CeHeChS) 4 parts 20.41 Active substance Microcrystalline 11 parts 56.12 Filler cellulose 102 Microcrystalline cellulose 12 1.5 parts 7.65 Filler Mannitol 3 parts 15.31 Filler Magnesium stearate 0.1 parts 0.51 Lubricant Empty gelatin capsule 1 capsule 1 capsule —

The resulting Compound 1 capsules were analyzed by high-performance liquid chromatography according to General Article 0941 Test Method for Dosage Uniformity of the Chinese Pharmacopoeia, 2015 Edition. The results are shown in Table. 23.

Table 23

Uniformity of dosing of capsules with compound 1

No. Value A+2.2S RSD, % Preparation 46 3.52 1.09 Preparation 47 3.21 0.86 Preparation 48 3.18 0.94

From the results of the study, it can be seen that the uniformity of dosing of drugs 46-48 was improved.

The resulting Compound 1 capsules were also subjected to a dissolution test using high performance liquid chromatography, and the results are shown in Table. 24.

- 29 045716

Table 24

Dissolution of capsules with compound 1______________

No. Dissolution rate % 0 month 3 min 5 minutes 10 min 15 minutes 20 minutes 30 min Preparation 46 66.24 91.21 99.81 100.03 101.08 101.15 Preparation 47 64.69 90.69 99.65 101.21 102.17 102.89 Preparation 48 71.30 90.29 96.46 98.30 98.98 99.06

From the results of the study, it can be seen that all the resulting compositions can be quickly dissolved.

Example 10 Stability test

Objectives: Conduct stability testing of drugs 47 and 48.

Method: Capsules of Drugs 47 and 48 were packaged in aluminum foil and rigid drug-grade PVC/PE/PVDC composite film, kept uncovered at 75% relative humidity and 40°C for 1 month, and then their impurity levels were measured. , active substance content (equal to the ratio of the tested active substance content to the labeled active substance content using the labeled content as the standard) and the dissolution rate. The results are shown in table. 25-27.

Table 25

Change in total impurity content

No. Total impurity content (%) 1 month 3 months 6 months Preparation 47 0.121 0.153 0.237 Preparation 48 0.117 0.136 0.197

Table 26

Change of content

No. Content, % (compared to content according to labeling) 1 month 3 months 6 months Preparation 47 100.57 100.16 99.98 Preparation 48 100.86 100.06 100.05

Table 27

Dissolution

No. Dissolution rate after 15 minutes (%) 1 month 3 months 6 months Preparation 47 99.74 99.12 98.31 Preparation 48 98.28 98.21 98.09

Example 11 Compound 1 tablets with different specifications.

Compound 1 tablets of various specifications (based on the free base of Compound 1) were prepared according to the composition given in Table. 28.

Table 28

Composition of tablets with compound 1

No. Starting materials Amount (mg) Functions Drug 49 (15 mg) Compound 1 (in terms of C12N19ХО2-SbNbOz8) 26.34 — Mannitol 42.31 Filler

- 30 045716

Microcrystalline cellulose 126.91 Filler Hydroxypropyl methylcellulose 15.00 Binder Croscarmellose sodium 20.00 Baking powder Magnesium stearate 2.00 Lubricant Drug 50 (30 mg) Compound 1 (in terms of CnHwNCh-CeHeCbS) 52.68 — Mannitol 42.31 Filler Microcrystalline cellulose 126.91 Filler Hydroxypropyl methylcellulose 15.00 Binder Croscarmellose sodium 20.00 Baking powder Magnesium stearate 2.00 Lubricant Preparation 51 (60 mg) Compound 1 (in terms of CnHwNCh-CeHeCbS) 105.36 — Mannitol 42.31 Filler Microcrystalline cellulose 126.91 Filler Hydroxypropyl methylcellulose 15.00 Binder Croscarmellose sodium 20.00 Baking powder Magnesium stearate 2.00 Lubricant Drug 52 (5 mg) Compound 1 (in terms of CnHwNCh-CeHeCbS) 8.78 Active substance Microcrystalline cellulose 102 114.14 Filler Microcrystalline cellulose 12 8.78 Filler Mannitol 26.34 Filler Magnesium stearate 0.88 Lubricant Drug 53 (5 mg) Compound 1 (in terms of CnHwNCh-CeHeCbS) 8.78 Active substance Microcrystalline cellulose 102 96.58 Filler Microcrystalline cellulose 12 8.78 Filler Mannitol 43.9 Filler Magnesium stearate 0.88 Lubricant Preparation 54 (20 mg) Compound 1 (in terms of С12Н19М)2<bНbОз8) 35.12 Active substance Microcrystalline cellulose 102 96.58 Filler Microcrystalline 13.17 Filler cellulose 12 Mannitol 26.34 Filler Magnesium stearate 0.88 Lubricant

The manufacturing method is as follows:

1) passing the active pharmaceutical ingredient, compound 1, through a 100 mesh sieve and passing the excipient and disintegrant through a 60 mesh sieve for use;

2) adding the active pharmaceutical ingredient, compound 1, excipient and disintegrant to the high-speed wet granulator for uniform mixing;

3) adding a predetermined amount of hydroxypropyl methylcellulose solution as a binder to obtain a soft material;

4) passing the soft material through a 20 mesh screen to obtain particles;

5) carrying out dynamic drying of the resulting particles in a fluidized bed to bring the moisture content to less than 2%;

6) passing the dried particles through a 24 mesh sieve to granulate;

7) adding granules with a given amount of magnesium stearate to the mixer with multidirectional movement for uniform mixing;

8) pressing the finished mixed powders using a matrix of a suitable size into tablets.

-31 045716

Example 12: Stability test.

Tablets of drugs 49-54 were kept open at a temperature of 40±2°C and a relative humidity of 75±5%, respectively. They were then tested for the corresponding substances on days 7, 14 and 30. The test results are shown in table. 29.

Table 29

Test results for relevant substances

Examples Amount of impurities 0 day 7 days 14 days 30 days Preparation 49 4 5 5 8 Preparation 50 5 5 5 8 Preparation 51 5 5 5 8 Preparation 52 3 4 4 5 Preparation 53 3 4 4 4 Preparation 54 3 4 5 6 Examples Total impurity content (%) 0 day 7 days 14 days 30 days Preparation 49 0.037 0.056 0.060 0.085 Preparation 50 0.061 0.066 0.097 0.120 Preparation 51 0.054 0.055 0.064 0.173 Preparation 52 0.039 0.048 0.051 0.073 Preparation 53 0.036 0.048 0.054 0.087 Preparation 54 0.040 0.052 0.060 0.085

Conclusion: from the test results it can be seen that the increase in the content of impurities in preparations 49-54 is insignificant, and the preparations are relatively stable.

Example 13 Preparation of tablets by one-step method.

Formulations 49-2, 50-2 and 51-2, respectively, were prepared using a top-spray fluidized bed according to the formulations of Formulations 49, 50 and 51.

The one-step method for preparing tablets is as follows:

1) passing the active pharmaceutical ingredient, compound 1, through a 100 mesh sieve and passing the excipient and disintegrant through a 60 mesh sieve for use;

2) adding the active pharmaceutical ingredient, Compound 1, excipient and disintegrant to a multi-directional mixer to ensure uniform mixing;

3) adding the mixed powder to the fluidized bed, supplying the hydroxypropyl methylcellulose solution to the fluidized bed through a peristaltic pump, and obtaining particles through overhead spraying, where the inlet temperature is set to 60 to 65°C, the temperature of the substances is controlled to 30 to 35°C, air supply speed is from 140 to 180 m ³ / h, and the peristaltic pump is adjusted at a level from 20 to 80 rpm;

4) passing the dried particles through a 24 mesh sieve to granulate;

5) adding granules with magnesium stearate to the mixer with multi-directional movement for uniform mixing; And

6) pressing the finished mixed powders using a matrix of a suitable size into tablets.

Tablets of drugs 49-2, 50-2 and 51-2 were kept open at a temperature of 40±2°C and a relative humidity of 75±5%, respectively. They were then tested for the corresponding substances on days 7, 14 and 30. The test results are shown in table. thirty.

- 32 045716

Table 30

Test results for relevant substances

No. Amount of impurities 0 day 7 days 14 days 30 days Preparation 49-2 4 5 5 8 Preparation 50-2 5 5 5 8 Preparation 51-2 5 5 5 8 No. Total impurity content (%) 0 day 7 days 14 days 30 days Preparation 49-2 0.036 0.055 0.060 0.084 Preparation 50-2 0.060 0.064 0.095 0.120 Preparation 51-2 0.053 0.054 0.064 0.172

Conclusion: From the test results, it can be seen that the increase in impurity levels in formulations 54-2, 56-2 and 62-2 is negligible and the tablets are relatively stable. However, when compared with tablets produced using a highly efficient wet granulation process, there is no noticeable difference in the impurity content.

Example 14. Biological analysis.

1. Study of competitive binding of compounds with the Cavα2δ protein voltage-dependent calcium channel.

Rat cortical tissue was placed in an ice-cold mixture of 0.32 M sucrose/5 mM Tris (trihydroxymethylaminomethane)-acetic acid (pH 7.4), having a 10-fold volume, and subjected to sucrose density gradient centrifugation to obtain synaptic plasmatic acid. membrane, which was stored in Tris-acetic acid buffer (pH 7.4) and resuspended in 10 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer (pH 7.4) before use. The test compound was dissolved in 1% DMSO (dimethyl sulfoxide) and diluted in a concentration gradient (1 nM-1000 nM), added to a suspension of synaptic plasma membrane (containing a total of about 0.05-0.1 mg protein) with 20 nM [ ^3H ]-gabapentin and incubated at 25°C for 30 min. At the end of the reaction, the system was vacuum filtered through Whatman GFB membranes, which were washed three times with 5 ml of 100 mM ice-cold sodium chloride. The radioactivity of the membranes was measured using a liquid scintillation counter. Nonspecific binding was blocked by 100 M gabapentin. The degree of inhibition by the compound of the binding of radiolabeled gabapentin to the synaptic plasma membrane was calculated, and the IC ₅₀ of the compound was calculated. The IC ₅₀ for compound 1 is 3.96 nM. Compound 1 has a good ability to competitively bind to the voltage-gated calcium channel protein Cava25.

2. Animal models with spinal nerve ligation L5-L6 (SNL).

Six- to seven-week-old male SD rats (from Charles River) were anesthetized with 5% isoflurane under animal conditions. Anesthetized animals were placed in a prone position and an incision was made at the level of the 5th lumbar vertebra. The skin was opened to expose the left paravertebral muscle, and the L5 and L6 spinal nerves were exposed by cutting layer by layer. 4-0 surgical wire was used to ligate the distal L5 and L6 dorsal root ganglia. The muscle and skin were stitched together layer by layer, and the animals were left to recover for one week.

After recovery, the model animals were tested for pain upon touch using a von Frey filament (DanMic Global; USA). The up-down method was used to measure the force applied to animals at 50% paw withdrawal response (g; 50% PWT). First of all, a group of animals was selected that responded to a force of 1-5 g at 50% PWT. Prior to administration of the compounds, animals were tested on baseline force levels, after which various compounds (mixed with 5% sodium carboxymethylcellulose) were orally administered, and the animals' pain response was tested at various time points over a testing range of 1.0 to 15 g. The test results are shown in Fig. 1.

Conclusion: The compounds of the present invention can significantly suppress the increased sensitivity to mechanical pain caused by spinal nerve ligation in rats.

3. Rat model of CCI-induced nociceptive sensitization.

SD rats weighing 160–180 g were purchased from Charles River (Beijing) with license number sCXK (Beijing)-2012-001 and qualification number 11400700254251.

To create the CCI model, surgery was performed as follows: animals were first anesthetized and then the left posterior sciatic nerve was exposed. A 2 mm long PE-90 catheter was inserted into the middle portion of the sciatic nerve. Finally, the muscle and skin were sutured. Adaptation of the left rear

- 33 045716 animal limbs for 15 g were carried out starting from 10 days after surgery 3 times a week.

Initial testing was performed 17 days after surgery (starting at 2 g at the time of testing, with an upper limit of 8 g and a lower limit of 1 g representing the -24 h time point) with a pain threshold of 1–4 g as the grouping criterion. Grouping was carried out in accordance with the results of the initial test, 10 animals per group. The animals in the group were not given food, but were given water at night. On the second day, mechanical pain was assessed at 2, 4 and 6 hours after oral/intragastric administration. The trial started at 8 g; with an upper limit of 15 g and a lower limit of 1 g.

Data were processed according to equation (1) to calculate a 50% PWT value, where Xf is the logarithmic value of the last tested fibers used in the test, k is the tabulated value (Chaplan et al., 1994), and δ is the mean difference ( here 0.224).

PWT 50%(g) = (10[Xf+k5])/10000 [Equation (1)]

Data were expressed as mean ± standard deviation (X±s) and statistically analyzed using one-way ANOVA and Dunnett's test (Graphpad prism 6.01).

The experimental results presented in Fig. 2 and 3 show that the mechanical pain threshold in SD rats with simulated CCI was significantly increased after a single oral administration of compound 1 and pregabalin. Compound 1, which has an effective dose of 3 mg/kg, is superior to pregabalin, which has an effective dose of 10 mg/kg.

Although specific embodiments of the invention have been described, those skilled in the art will understand that various changes and modifications may be made to the invention without departing from the scope or spirit of the invention. Accordingly, the present invention is intended to cover all such changes and modifications that come within the scope of the appended claims and their equivalents.

Claims (7)

1. Pharmaceutical composition containing: (1) an active substance having the structure represented by formula (I) or a pharmaceutically acceptable salt thereof in an amount of 1 to 45% by weight; (2) a filler in an amount of from 50 to 95 wt.%, containing mannitol in an amount from 9 to 35 wt.%, microcrystalline cellulose A in an amount from 3 to 60 wt.% and microcrystalline cellulose B in an amount from 17 to 72 wt. %, where microcrystalline cellulose A is selected from the group consisting of microcrystalline cellulose PH14, microcrystalline cellulose PH12, microcrystalline cellulose 12 and microcrystalline cellulose 14; microcrystalline cellulose B is selected from the group consisting of microcrystalline celluloses PH102, PH105, PH103, PH301, PH101, PH112, PH302 and microcrystalline celluloses 102, 105, 103, 301, 101, 112, 302; and (3) one or more lubricants in an amount of from 0.1 to 5.5 wt.%; where the sum of the mass percentages of all components is 100%; where formula (I) has the following structure: n _r ^NH 2 lf,co ₂ H 2. The composition according to claim 1, where formula (I) has the following structure: H / ^NH 2 HC...../CO ₂ H 3. The composition according to claim 2, wherein the pharmaceutically acceptable salt of the compound of formula (I) has the following structure: 4. Composition according to claim 1, where the active substance is included in an amount of from 1 to 25 wt.%. 5. The composition according to claim 1, where the microcrystalline cellulose contains microcrystalline cellulose 102 and microcrystalline cellulose 12. 6. The composition according to claim 1, wherein the lubricant is one or more lubricants selected from stearic acid, magnesium stearate, sodium stearyl fumarate and glycerol behenate, preferably magnesium stearate. - 34 045716 7. The composition according to claim 6, where the lubricant is included in an amount of from 0.5 to 2 wt.%. 8. The composition according to claim 1, further containing one or more than one binder, which is selected from hydroxypropylcellulose or povidone. 9. The composition of claim 1, further comprising one or more disintegrants selected from croscarmellose sodium and sodium carboxymethyl starch. 10. The composition of claim 1, further comprising one or more glidants selected from silica or talc. 11. The composition according to claim 1, which is in the form of a capsule or tablet. 12. Composition according to claim 11, containing: (1) a pharmaceutically acceptable salt of the compound of formula (I) as an active substance, which is included in an amount of 5 to 21% by weight; (2) a filler containing mannitol, microcrystalline cellulose A and microcrystalline cellulose B, wherein mannitol is included in an amount of 15 to 28 wt.%, microcrystalline cellulose A is included in an amount of 5 to 8 wt.% and microcrystalline cellulose B is included in an amount of from 56 to 61 wt.%; and (3) magnesium stearate, which is included in an amount of 0.5 to 0.6% by weight; wherein the total percentage of all components is 100% and the pharmaceutically acceptable salt of the compound of formula (I) has the following structure: 13. The composition according to claim 12, further containing a glidant selected from silicon dioxide or talc. 14. The composition according to claim 12, further containing a binder selected from hydroxypropylcellulose. 15. Composition according to any one of claims 1 to 14, wherein the active substance is present, in terms of its free base, in an amount of from 1 to 100 mg, preferably in an amount of from 5 to 20 mg. 16. Composition according to any one of claims 1 to 14, where the active substance is present, in terms of its free base, in an amount of 5 or 20 mg. 17. A method for producing a pharmaceutical composition according to any one of claims 1-14 in the form of a capsule, including: 1) passing the active substances through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use; 2) adding the sifted active substance and filler to the mixer with multidirectional movement for uniform mixing; 3) adding lubricant to the multi-directional mixer to mix with the mixture obtained in step (2); And 4) filling the capsule with the mixture obtained in step (3). 18. A method for manufacturing a pharmaceutical composition according to any one of claims 1-14 in the form of a capsule, including: 1) passing the active substances through a 60 mesh sieve and passing the excipient through a 40 mesh sieve for use; 2) preparing a 40% aqueous solution of ethanol as a solvent; 3) adding the sieved active substance and filler with the binder to the wet granulator and adding the solvent obtained in step (2) for shear mixing to obtain a soft material; 4) passing the soft material through a sieve to obtain particles, and performing static drying or dynamic fluidized bed drying; 5) granulating the dried soft material and then adding it with a glidant and disintegrant to a multi-directional mixer for uniform mixing; 6) adding lubricant to the multi-directional mixer for mixing; 7) filling the capsule with the mixture obtained in step (6); or alternatively, steps (2) and (3) may be replaced by: 2-1) preparation of a binder and a 40% aqueous solution of ethanol in the form of a binder solution; 3-1) adding the active substance and filler to the wet granulator and adding the binder solution obtained in step (2-1) for shear mixing to obtain a soft material. - 35 045716 19. A method for producing a pharmaceutical composition according to any one of claims 1-14 in the form of a tablet, including: 1) passing the active ingredients through a 100 mesh sieve and passing the filler and disintegrant through a 60 mesh sieve; 2) adding the sieved active ingredient, filler and disintegrant into the high-speed wet granulator to ensure uniform mixing; 3) adding a binding solvent to the mixed powder obtained in step (2) to obtain a soft material; 4) passing the soft material through a 20 mesh screen to obtain particles, then subjecting it to static drying or dynamic fluid bed drying to reduce the moisture content to less than 2%; 5) passing the dried particles through a 24 mesh sieve for granulation, adding lubricant granules to the mixer with multi-directional motion to ensure uniform mixing; 6) pressing the mixture obtained in step (5) into tablet form; alternatively, steps (3) and (4) can be replaced by: (3-1) adding the mixed powder obtained in step (2) to the fluidized bed, supplying the binder solution to the fluidized bed by a peristaltic pump, and producing particles by overhead spraying. 20. Use of a pharmaceutical composition according to any one of claims 1 to 14 in the preparation of a medicinal product for the treatment and/or prevention of mechanical pain.