Sterile bis(tapentadol) sebacate, preparation method therefor and use thereof

Abstract

The present invention relates to sterile bis(tapentadol) sebacate, a preparation method therefor, and the pharmaceutical use of a pharmaceutical preparation containing same. Specifically, the sterile bis(tapentadol) sebacate has a particle size D50 of 0.5-10 μm. Further provided in the present invention is the use of the bis(tapentadol) sebacate in the preparation of various drugs for acute and chronic pain, which has significant long-acting characteristics.

Description

A sterile bis(tapeptide) dodecyl ester, its preparation method and uses Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a sterile bis(tapeptide) decanterate, its preparation method, and its pharmaceutical formulation and medicinal uses. Background Technology

[0002] Most patients experiencing moderate to severe pain, such as postoperative pain, post-traumatic pain, and burn pain, typically require pain control within the first three days after injury. Therefore, analgesics with a duration of action of approximately three days may be particularly valuable (KS. Chu, et al. (2003), Anesthesia Analgesia, Vol. 97, 806-809). Currently, opioids and nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used in this field, but they are all short-acting. Extending the duration of action (e.g., long-acting opioids) would be more valuable in the clinical treatment of long-term pain. In recent years, prolonging the duration of analgesia and reducing the frequency of postoperative opioid use have gradually become clinical analgesic needs; therefore, the application of long-acting sustained-release analgesics postoperatively has become a research hotspot.

[0003] Pain is an unpleasant sensory and emotional experience associated with, or described in relation to, actual or potential tissue damage. Pain can be classified by etiology, duration, and severity. Etiologically, pain can be classified as somatic (i.e., organic) or psychogenic (occurring in the absence of a relevant organic pathology sufficient to explain the severity and / or duration of the pain). Somatic pain can be further subdivided into nociceptive pain (caused by stimulation of pain-sensitive nerve fibers in the body or viscera) or neurogenic pain (caused by dysfunction of the nervous system). Regarding duration, pain is generally classified as acute or chronic. Chronic persistent pain can lead to serious impairment of physical and mental health and social responsibilities (including work and family life). Chronic pain is described as pain lasting from at least 5 days to up to 6 months. Chronic pain is often associated with conditions such as surgery, cancer, and serious injury. Opioids are commonly used to control severe chronic pain conditions. Although opioids are potent analgesics, their relatively short half-life limits their benefits to some extent. Because the pain of the procedure can last for several days, these analgesics must be administered multiple times to effectively control the pain.

[0004] Among opioid dual analgesics, tapenetadol is the most effective. The chemical name of tapenetadol is 3-((1R,2R)-3-(dimethylamino)-1-ethyl-2-methylpropyl)phenol, with a molecular weight of 221.34, represented by the following structural formula:

[0005] Tapentadal is an opioid analgesic with μ-opioid receptor agonist and norepinephrine reuptake inhibitory activity, with minimal serotonin reuptake inhibition. This dual mode of action makes Tapentadal particularly useful in treating nociceptive and neuropathic pain. Clinical trial evidence in acute and chronic non-cancer pain and neuropathic pain supports the opioid-reducing effect, thereby reducing some typical opioid-related adverse reactions. Specifically, Tapentadal treatment results in fewer gastrointestinal adverse reactions compared to equivalent pure μ-opioid receptor agonists, thus improving tolerability and treatment adherence.

[0006] U.S. Patent No. 6,248,737 discloses Tapental and its hydrochloride. Tapental is marketed under the trade name... It is available on the market as an immediate-release oral tablet for the relief of moderate to severe acute pain. RETARD, as a sustained-release tablet, is used for severe chronic pain.

[0007] When tapentadal is taken orally, it undergoes extensive first-pass metabolism, resulting in low bioavailability (32%). Approximately 97% of the parent compound is metabolized. No metabolites contribute to the analgesic activity. Ultimately, only high doses of tapentadal can achieve the desired effect. Immediate-release oral tapentadal is administered every 4–6 hours, while extended-release tablets are administered every 12 hours. As an opioid analgesic, tapentadal is used to treat severe pain, such as postoperative pain, cancer pain, etc. In such cases, nausea and vomiting are frequently associated problems, leading to poor patient adherence to oral administration. Some disadvantages of oral administration include the possibility that patients may inappropriately modify the unit dose, resulting in a dangerous overdose, or that patients may be unable to swallow the medication.

[0008] Tapental's short duration of action forces patients to take it frequently. Furthermore, like other opioids, tapental is considered to have the potential for abuse. To address this issue, U.S. Patent No. 8,075,872 provides an abuse-preventing controlled-release formulation of tapental for oral administration, twice daily.

[0009] Long-lasting analgesia is particularly desirable for patients suffering from pain, such as postoperative pain, post-traumatic pain, and burn pain, which may last for about 3 days. Tadalafil has a strong but short-lived analgesic activity. Extending the duration of action would make Tadalafil more valuable in the clinical treatment of pain.

[0010] Therefore, an alternative dosage form is needed that provides a longer-acting tapentam, thereby reducing the frequency of administration. Furthermore, alternative dosage forms are needed to overcome the problems associated with oral administration and reduce opportunities for abuse, so that the release of the analgesic cannot be manipulated by the patient or other external sources.

[0011] Patent documents:

[0012] US20130090379, Compositions comprising an enzyme-cleaved phenol-modified tapental prodrug.

[0013] US20100227921, Tapentadole amino acid and peptide carbamate prodrugs and their uses

[0014] JP202307524, He sprayed more parenteral medication.

[0015] US10898452, a stable formulation of tadalafil for parenteral administration.

[0016] CN2014071658, A tapentathol hydrochloride injection solution and its preparation method

[0017] Several tapentadol prodrugs have been synthesized and reported. However, no known tapentadol ester has been used for long-acting purposes. Therefore, there remains a need in the art to develop a suitable pharmaceutical composition that allows tapentadol to exhibit long-acting analgesic effects. Summary of the Invention

[0018] This invention modifies the structure of tapentadine to prepare a prodrug with long-acting properties. This type of drug is formulated into a preparation suitable for intramuscular, subcutaneous, or intravenous injection. After intramuscular, subcutaneous, or intravenous injection, it forms a drug reservoir in the body, from which the drug is slowly, continuously, and stably released and converted into tapentadine, thereby exerting a long-acting effect. This invention is achieved using the following technical solution:

[0019] The present invention addresses the shortcomings of the prior art by providing a compound represented by formula (Ⅰ) (i.e., bis(tapenta) decanterate):

[0020] Preferably, the compound of the present invention (bisapenta dodecanedioate) refers to a sterile active pharmaceutical ingredient, and more preferably, the particle size D of the bisapenta dodecanedioate raw material is... 50 The range is 0.5–10 μm.

[0021] Another aspect of the present invention is to provide a crystal form of bis(tapeptide) dodecyl ester that is free of water of crystallization or solvent.

[0022] Preferably, the PXRD pattern of the crystal form has characteristic peaks selected from the following, located at 7.82, 13.56, 15.60, 16.30 and 16.72, wherein the characteristic peaks are represented by CuKα radiation at 2θ ± 0.2°2θ.

[0023] Preferably, the PXRD pattern of the crystal form has characteristic peaks selected from the following, located at 7.82, 13.54, 13.56, 14.34, 15.60, 16.14, 16.30, 16.68 and 16.72, wherein the characteristic peaks are represented by CuKα radiation at 2θ ± 0.2°2θ.

[0024] The preferred crystal form has a PXRD pattern with characteristic peaks selected from the following: 7.82, 13.54, 13.56, 14.34, 15.60, 16.14, 16.30, 16.68, 16.72, 17.34, 17.82, 17.86, 18.56, 19.60, 20.70, 21.24, 22.12, and 22.80, wherein the characteristic peaks are represented by CuKα radiation at 2θ ± 0.2°2θ.

[0025] Preferably, the PXRD pattern of the crystal form has characteristic peaks selected from the following: 7.82, 13.54, 13.56, 14.34, 15.60, 16.14, 16.30, 16.68, 16.72, 17.34, 17.82, 17.86, 18.56, 19.60, 20.70, 21.24, 22.12, and 22.80, wherein the characteristic peaks are represented by CuKα radiation at 2θ ± 0.2°2θ.

[0026] Preferably, the PXRD pattern of the crystal form has characteristic peaks selected from the following: 7.82, 13.54, 13.56, 14.34, 15.60, 16.14, 16.30, 16.68, 16.72, 17.34, 17.82, 17.86, 18.56, 19.60, 20.70, 21.24, 22.12, 22.80, 23.60, 24.04, 24.24, 25.00, 25.12, and 27.50, wherein the characteristic peaks are represented by CuKα radiation at 2θ ± 0.2°2θ.

[0027] The PXRD pattern of the preferred crystal form is shown in Figure 7.

[0028] The preferred crystal form of TGA shows decomposition starting at approximately 250°C;

[0029] Preferably, the DSC reading of the crystal form shows a melting point of 51.90°C.

[0030] Another aspect of the present invention provides a method for preparing bis(tapeptadol) sebacic acid ester, comprising the step of esterification of bis(tapeptadol) hydrochloride with sebacic acid chloride in an organic solvent and an alkali-binding acid agent, the reaction formula being as follows:

[0031] The preparation of tapentathol hydrochloride was carried out in accordance with the Chinese Journal of Medicine, 2013, 44(6): 554-556.

[0032] Preferably, the above preparation method further includes a crystallization process.

[0033] A series of crystallization methods were screened during the preparation of the bis(tapeptide) polysaccharide crystal form, and the results are as follows:

[0034] Table 1 Screening of crystallization solvents and methods

[0035] The solvent used in the crystallization process provided by the present invention is selected from one or more of methyl tert-butyl ether, isopropyl ether, petroleum ether, n-hexane, cyclohexane, and n-pentane; preferably one or more of methyl tert-butyl ether, isopropyl ether, n-hexane, and petroleum ether.

[0036] The crystallization process provided by the present invention has a temperature of -15 to 5°C, preferably -15 to 0°C.

[0037] Another aspect of the present invention is to provide a method for preparing a sterile bis(tapeptide) dodecyl ester, which includes pre-filtration with a 0.45 μm microporous membrane before the crystallization process, followed by two stages of sterile filtration with a 0.22 μm membrane.

[0038] Another aspect of the present invention is to provide a pharmaceutical composition of bis(tapeptone) dodecanoate, the pharmaceutical composition comprising bis(tapeptone) dodecanoate and pharmaceutically usable excipients.

[0039] The preferred route of administration is injection; the preferred dosage form is injection solution, lyophilized powder, etc.

[0040] Preferably, the injection solution is prepared from the active sterile raw material dicapentate sebacic acid and related pharmaceutical excipients according to general pharmaceutical methods. The excipients include pharmaceutically necessary components such as suspending agents, antioxidants, stabilizers, wetting agents, and preservatives.

[0041] Another aspect of the present invention is to provide a pharmaceutical composition of bis(tapeptide) dodecanoate or bis(tapeptide) dodecanoate in the preparation of various acute and chronic pain medications.

[0042] Another aspect of the present invention is to provide the use of a pharmaceutical composition of bis(tapeptone) dodecanoate or bis(tapeptone) dodecanoate in the treatment of various acute and chronic pain disorders.

[0043] The key points of this invention are:

[0044] The present invention relates to bis(tapeta)dextrose dodecanoate and its crystalline form, which exhibit good stability and are suitable for storage. Aseptic bis(tapeta)dextrose can be prepared by a simple method, making it suitable for industrial production. Furthermore, bis(tapeta)dextrose can be formulated into a suspension for intramuscular or subcutaneous injection, forming a drug reservoir in the body and prolonging the drug release rate, thus achieving a long-lasting therapeutic effect. Moreover, this compound has virtually no toxicity to normal human hepatocytes. Animal pharmacodynamic and safety experiments show good efficacy, a sustained analgesic effect, and no significant irritation to local tissues, demonstrating good safety and tolerability. Animal pharmacokinetic experiments show that the compound has a long half-life, achieving a long-lasting release effect. Attached Figure Description

[0045] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0046] Figure 1 shows the compound structure of bis(tapeptide) dodecyl ester.

[0047] Figure 2 shows the hydrogen spectrum of bis(tapeptide) decanterate.

[0048] Figure 3 shows the carbon spectrum of bis(tapeptide) decanterate.

[0049] Figure 4 shows the DSC of bis(tapeptide) decanterate.

[0050] Figure 5 shows the IR of bis(tapeptide) dodecyl ester.

[0051] Figure 6 shows the results of the analgesic pharmacodynamic experiment of distatin dodecadiate in rats.

[0052] Figure 7 shows the powder X-ray diffraction pattern of bis(tapeptide) polysaccharide. Detailed Implementation

[0053] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0054] The present invention will be further explained below with reference to specific embodiments.

[0055] Example 1:

[0056] 400 mL of dichloromethane and 25.70 g of tapentadone hydrochloride were added to a 1000 mL three-necked flask. 20.25 g of triethylamine was added with stirring. After several minutes, the mixture was completely dissolved. The temperature was lowered to -5 to 0 °C, and 12.00 g of sebacyl chloride was added dropwise. The temperature rose significantly. The internal temperature was controlled at 0–10 °C, and insoluble substances gradually precipitated out. The reaction was stirred at 0–10 °C for 2 hours. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed that the tapentadone content was ≤0.5%. 150 mL of water was added to quench the reaction. After separation, the mixture was washed twice with 400 mL of water. The dichloromethane layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 20.70 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became a white solid. HPLC analysis showed a purity of 97.68%.

[0057] 1 H-NMR (400MHz, DMSO-d6): δ [ppm] = 7.339-7.300 (t, 2H), 7.047-7.028 (d, 2H), 6.938-6.883 (m, 4H), 2.585-2.549 (t, 4H), 2.445-2.394 (m, 2H), 2 .043(s, 12H), 1.955-1.914(m, 2H), 1.886-1.788(m, 4H), 1.758-1.525( m, 8H), 1.447-1.355 (m, 8H), 0.876-0.860 (d, 6H), 0.687-0.651 (t, 6H).

[0058] Example 2:

[0059] 800 mL of dichloromethane and 51.40 g of terbutaline hydrochloride were added to a 2000 mL three-necked flask. 40.50 g of triethylamine was added with stirring. After several minutes, the mixture was completely dissolved. The temperature was lowered to -5 to 0 °C, and 24.00 g of sebacyl chloride was added dropwise. The temperature rose significantly. The internal temperature was controlled at 0–10 °C, and insoluble substances gradually precipitated out. The reaction was stirred at 0–10 °C for 3 hours. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed terbutaline concentration ≤ 0.5%. 300 mL of water was added to quench the reaction. After separation, the mixture was washed twice with 800 mL of water. The dichloromethane layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 42.40 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became a white solid.

[0060] Example 3:

[0061] 1500 mL of dichloromethane and 102.80 g of terbutaline hydrochloride were added to a 3000 mL three-necked flask. 81.00 g of triethylamine was added with stirring. After several minutes, the mixture was completely dissolved. The temperature was lowered to -5 to 0 °C, and 47.80 g of sebacyl chloride was added dropwise. The temperature rose significantly. The internal temperature was controlled at 0–10 °C, and insoluble substances gradually precipitated out. The reaction was stirred at 0–10 °C for 4 hours. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed terbutaline concentration ≤ 0.5%. 600 mL of water was added to quench the reaction. After separation, the mixture was washed twice with 1500 mL of water. The dichloromethane layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 80.00 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became a white solid.

[0062] Example 4:

[0063] 400 mL of redistilled toluene and 25.70 g of terpentine hydrochloride were added to a 1000 mL three-necked flask. 20.25 g of triethylamine was added with stirring. The mixture did not dissolve completely. The temperature was lowered to -5 to 0 °C, and 12.00 g of sebacyl chloride was added dropwise. The temperature rose significantly. The reaction was stirred for 2 hours while maintaining the internal temperature at 0 to 10 °C. TLC monitoring was performed (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added). Terpentine concentration was ≤0.5%. 150 mL of water was added to quench the reaction. After separation, the organic phase was washed twice with 400 mL of water. The toluene layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 21.50 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became a white solid.

[0064] Example 5:

[0065] 130 mL of dry N,N-dimethylformamide and 25.70 g of tapentadone hydrochloride were added to a 500 mL three-necked flask. 20.25 g of triethylamine was added with stirring. After stirring for several minutes, the mixture was completely dissolved. The temperature was lowered to -5 to 0 °C, and 12.00 g of sebacyl chloride was added dropwise. The temperature rose significantly. The reaction was stirred for 2 hours while maintaining the internal temperature at 0 to 10 °C. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed tapentadone concentration ≤ 0.5%. The reaction was quenched with 150 mL of water. The mixture was extracted with 500 mL of ethyl acetate to separate the layers. After separation, the organic phase was washed twice with 400 mL of water. The ethyl acetate layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 18.60 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became a white solid.

[0066] Example 6:

[0067] 150 mL of dry tetrahydrofuran and 25.70 g of terpentine hydrochloride were added to a 500 mL three-necked flask. 20.25 g of triethylamine was added while stirring. After stirring for several minutes, the material was not completely dissolved. The temperature was lowered to -5 to 0 °C, and 12.00 g of sebacyl chloride was added dropwise. The temperature rose significantly. The internal temperature was controlled at 0 to 10 °C and the reaction was stirred for 2 hours. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed terpentine ≤ 0.5%. 150 mL of water was added to quench the reaction. The mixture was extracted with 500 mL of ethyl acetate to form layers. After separation, the organic phase was washed twice with 400 mL of water. The ethyl acetate layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 19.50 g of a pale yellow oily substance. After being placed in the refrigerator overnight, it became a white solid.

[0068] Example 7:

[0069] 400 mL of dichloromethane and 25.70 g of terbutaline hydrochloride were added to a 1000 mL three-necked flask. 20.25 g of triethylamine was added with stirring. After several minutes, the mixture was completely dissolved. The temperature was lowered to -5 to 0 °C, and 23.90 g of sebacyl chloride was added dropwise. The temperature rose significantly. The internal temperature was controlled at 0–10 °C, and insoluble substances gradually precipitated out. The reaction was stirred at 0–10 °C for 2 hours. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed terbutaline concentration ≤ 0.5%. 500 mL of water was added to quench the reaction. After separation, the mixture was washed twice with 500 mL of water. The dichloromethane layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 19.30 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became a white solid.

[0070] Example 8:

[0071] 400 mL of dichloromethane and 25.70 g of terbutaline hydrochloride were added to a 1000 mL three-necked flask. 20.25 g of triethylamine was added with stirring. After several minutes, the mixture was completely dissolved. The temperature was lowered to -5 to 0 °C, and 18.00 g of sebacyl chloride was added dropwise. The temperature rose significantly. The internal temperature was controlled at 0 to 10 °C, and insoluble substances gradually precipitated out. The reaction was stirred at 0 to 10 °C for 2 hours. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed terbutaline concentration ≤ 0.5%. 400 mL of water was added to quench the reaction. After separation, the mixture was washed twice with 400 mL of water. The dichloromethane layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 20.30 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became a white solid.

[0072] Example 9:

[0073] Take 1.00 g of the white solid obtained in Example 1, add 2 mL of methyl tert-butyl ether and 5 mL of isopropyl ether, stir and heat until completely dissolved, add an appropriate amount of activated carbon for needles and keep warm for 15 min, pre-filter using a 0.45 μm microporous membrane, then pass through two stages of 0.22 μm sterile filtration, then slowly cool to 15-25 °C, and continue cooling to -5-0 °C, and keep at -5-0 °C for 3 days to crystallize, filter the obtained solid under reduced pressure, and vacuum dry at 40 °C to obtain 0.89 g of white solid. HPLC detection result: 98.59%, sent for infrared, TGA and DSC detection.

[0074] Example 10:

[0075] Take 1.00 g of the white solid obtained in Example 1, add 2 mL of methyl tert-butyl ether, stir and heat until completely dissolved, add 5 mL of n-hexane during reflux, add an appropriate amount of activated carbon for needle insertion and keep warm for 15 min, pre-filter using a 0.45 μm microporous membrane, then pass through two stages of 0.22 μm sterile filtration, then slowly cool to 15–25 °C, and continue cooling to -5–0 °C, and keep at -5–0 °C for 3 days to crystallize, filter the obtained solid under reduced pressure, and vacuum dry at 40 °C to obtain 0.91 g of white solid, the HPLC detection result is: 98.07%.

[0076] Example 11:

[0077] Take 1.00 g of the white solid obtained in Example 1, add 7 mL of methyl tert-butyl ether, stir and heat until completely dissolved, add an appropriate amount of activated carbon for injection and keep warm for 15 min, pre-filter using a 0.45 μm microporous membrane, then pass through two stages of 0.22 μm sterile filtration, then slowly cool to 15-25 °C, and continue cooling to -5-0 °C, and keep at -5-0 °C for 3 days to crystallize, filter the obtained solid under reduced pressure, and vacuum dry at 40 °C to obtain 0.86 g of white solid. HPLC detection result: 96.13%.

[0078] Table 2. Detection results of the compounds of this invention by 1H NMR.

[0079] Table 3. Detection results of the compounds of this invention by carbon NMR.

[0080] Table 4 Mass spectrometry detection results of the compounds of this invention

[0081] Table 5 Infrared detection results of the compounds of this invention

[0082] TGA test results of the compound of this invention: As can be seen from the TGA graph, this product does not contain water of crystallization or solvent and begins to decompose at about 250°C.

[0083] DSC test results of the compound of this invention: As can be seen from the DSC graph, this product does not contain water of crystallization or crystallization solvent, and the melting point is 51.90℃.

[0084] Table 6. PXRD detection results of the compounds of this invention.

[0085] Example 12: Preparation of bis(tapeptadol) nonanoate and bis(tapeptadol) undecanoate

[0086] Referring to the synthesis method of Example 1, sebacate chloride was replaced with azelaate chloride or undecyl diacyl chloride to synthesize bis(tapeptadol) nonacidate and bis(tapeptadol) undecylate.

[0087] Table 7. Mass spectrometry and appearance of bis(tapeptide) nonaconate and bis(tapeptide) undecanoate.

[0088] Example 13: Preparation of monotabentatate polydecanoate

[0089] 400 mL of dichloromethane and 25.70 g of tapentathione hydrochloride were added to a 1000 mL three-necked flask. While stirring, 20.25 g of triethylamine, 4.5 g of DCC, and 0.4 g of DMAP were added. After several minutes, the mixture was completely dissolved. The temperature was lowered to -5 to 0 °C, and 19.3 g of sebacic acid was added dropwise. The temperature rose significantly. The internal temperature was controlled at 0 to 10 °C, and insoluble substances gradually precipitated out. The reaction was stirred at 0 to 10 °C for 2 hours. TLC monitoring (λ = 254 nm, PE:EA = 2:1, two drops of triethylamine added) showed that the tapentathione content was ≤ 0.5%. 100 mL of water was added to quench the reaction. After separation, the mixture was washed twice with 400 mL of water. The dichloromethane layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 13.01 g of a pale yellow oily substance. After being placed in a refrigerator overnight, it became an amorphous powder solid.

[0090] Table 8. Mass spectrometry and appearance of monocapentatone polydecanoate.

[0091] Example 14: Particle size distribution detection

[0092] The particle size of the active pharmaceutical ingredient was determined according to Method 3, General Chapter 0982, Part IV of the 2020 edition of the Chinese Pharmacopoeia. Particle size distribution was analyzed for the products of Examples 9, 10, and 11. Specific procedure: An appropriate amount of the crystalline powder from Examples 9, 10, and 11 was placed in a 5 mL centrifuge tube, distilled water was added, and the mixture was shaken well. A background scan was performed against a background of distilled water to determine the background value. Then, an appropriate amount of the test solution was placed in the sample cell, and its particle size distribution was analyzed. The results are as follows:

[0093] Table 9. Particle size distribution detection results of the compounds of this invention.

[0094] Example 15: Stability Test

[0095] The compounds of this invention will be used as active pharmaceutical ingredients (APIs) in the preparation of suspension injections; therefore, storage is crucial. Impurities may be introduced or generated during storage, affecting the use of the API. Therefore, the stability of the API of this invention was investigated. Referring to the ICH guideline "Stability Testing of New APIs and Formulations," accelerated testing was conducted to investigate the effects of temperature, humidity, and light on the APIs prepared in Examples 9, 10, and 11. The conditions were set as follows: temperature 40 ± 2 °C, humidity 75% ± 5%, light intensity 4500 lx, and sampling was performed after 10 days. The results are shown in the table below:

[0096] Table 10 Stability test results of the compounds of the present invention

[0097] Conclusion: As shown in the table, the active pharmaceutical ingredient of the present invention is relatively stable under the relevant conditions and no obvious impurities are generated. Therefore, after the active pharmaceutical ingredient is prepared, it can be stored at room temperature.

[0098] Example 16: Experimental study on in vitro cytotoxicity of normal human hepatocytes

[0099] 1. Experimental Materials

[0100] 1.1 Cells: LO2 cells, a human hepatitis cell line

[0101] 1.2 Drug: The compound of this invention has an HPLC purity of 98.07%.

[0102] 1.3 Reagents and Instruments: Modified RPMI-1640 culture medium, penicillin-streptomycin solution, 0.25% trypsin-EDTA, fetal bovine serum, MTT, dimethyl sulfoxide, CO-150 carbon monoxide incubator, SW-CJ-2F medical clean bench, CKX-41-32 inverted microscope, CU600 electric thermostatic water bath, RT-2100C enzyme-linked immunosorbent assay (ELISA) analyzer.

[0103] 2 Experimental Methods

[0104] 2.1 Reagent Preparation

[0105] 2.1.1 Preparation of MTT: Weigh 0.25g of MTT using a precision balance and place it in a 50mL volumetric flask. Add an appropriate amount of PBS, incubate in a 50-60℃ water bath, and shake well to dissolve completely. Add PBS to the mark to prepare a 5mg / kg solution. Filter the solution through a 0.22μm microporous membrane for sterilization, aliquot, and store in a refrigerator at 4℃ protected from light.

[0106] 2.1.2 Preparation of cell cryopreservation solution: Mix 20% serum, 10% DMSO and 70% 1640 medium evenly and store at -20℃.

[0107] 2.1.3 The compound of the present invention is prepared by using DMSO to prepare a stock solution of the drug, and then diluting it with culture medium to the concentration of the drug to be used. The final concentration of DMSO is controlled to be ≤0.1%.

[0108] 2.2 LO2 cell culture: Normal human LO2 cells were placed in a 25cm² culture medium. 2 Add approximately 4–5 mL of RPMI-1640 culture medium containing 10% FBS to cell culture flasks and incubate at 37°C in a 5% CO2 saturated humidity incubator. Change the culture medium every 2 days and observe cell growth daily. Once cells reach 80% confluence, passage or cryopreserve them. Use cells from passages 5–7 for formal experiments.

[0109] 2.3 Grouping and Drug Administration Experiment: The experiment was divided into a normal cell control group and drug administration groups with different concentrations of the present invention, based on the preliminary experimental results. The concentrations were 5.0, 10.0, 20.0, 40.0, 80.0, 160, 320, 640, 1280, and 1500 μmol / L.

[0110] 2.4 Hepatocyte MTT assay: Logarithmic growth phase LO2 cells were prepared into a cell suspension of 5.0 × 10³ cells / mL and seeded into 96-well plates. A normal control group and groups treated with different concentrations of the compound of the present invention (4.0, 8.0, 16.0, 32.0, 64.0, 128, 256, 512, 800, and 1000 μmol / L) were included. After 24 h of culture, the culture medium was discarded, and the cells were washed 2–3 times with PBS. Different concentrations of the compound of the present invention were added, with 8 replicates per concentration. The plates were incubated at 37°C in a 5% CO₂ incubator. After 24, 48, and 72 h of culture, MTT solution was added at each time point, and the plates were incubated in the dark for 4 h. After 4 h, the supernatant was discarded, and 150 LDMSO was added to each well to dissolve the thiazolyl blue crystals. The plates were gently shaken to ensure uniform dissolution. The absorbance of each well was measured at 490 nm using a microplate reader. The absorbance value per well reflects the number of cells, and the two are directly proportional. The experiment was repeated three times. Cell viability was calculated as follows: Cell viability (%) = Absorbance of each group (OD490) × 100 / Absorbance of the control group (OD490)

[0111] 2.5 Statistical Analysis All data are expressed as mean ± standard deviation and were processed using SPSS 17.0 statistical software. t-tests were performed for statistical analysis; paired t-tests were used for self-comparisons, and unpaired t-tests were used for inter-group comparisons. The significance level was P < 0.05.

[0112] 3 Experimental Results

[0113] Table 11. Experimental results of in vitro cytotoxicity of the compounds of the present invention on normal human hepatocytes.

[0114] Conclusion: After 72 hours of administration, the OD values ​​of cells in each group showed an increasing trend, but the growth rate decreased, indicating that the cells were still in the growth stage and the growth rate was somewhat inhibited. Different concentrations of the present invention showed no inhibitory effect on cells; compared with the normal group, the cell survival rate was greater than 90%, and within the range of 5.0–1500 μmol / L, it showed almost no inhibition of cell growth and no cytotoxicity.

[0115] Example 17: Analgesic Pharmacodynamic Evaluation

[0116] 1. Laboratory animals

[0117] Male Sprague-Dawley (SD) rats, weighing (200±20)g.

[0118] 2 Experimental Methods

[0119] This study investigated the efficacy of subcutaneous administration of the present invention in a rat dorsal incision model. Male SD rats (n=3) were used as experimental animals. All animals were acclimatized to their environment for 2-3 days prior to surgery, and particularly sensitive or sluggish animals were excluded. The selected rats were anesthetized with Supra-50. Under aseptic conditions, a 2cm longitudinal incision was made on the left side of the rat's dorsal midline, reaching the fascia layer without damaging the muscle. The fascia and muscle were separated, and the skin was discontinuously sutured at the incision site. The drug was administered at a dose of 1.5 mL / kg. -1 Subcutaneous injection of the hydrogenated castor oil solution of the present invention (prepared by dissolving the compound of the present invention in hydrogenated castor oil), (2.5 mg·kg) -1 5mg·kg -1 10 mg·kg -1 The incision sites were divided into low, medium, and high dose groups, with an untreated incision group serving as a blank control. Before administration and at 1, 2, 3, 4, 8, 12, 24, 36, 48, 60, 72, and 84 hours after administration, the area near the incision site was stimulated with different Von-frey fibers (providing tactile stimulation force of 0.008–300 g). The pain threshold was determined by the g value represented by the Von-frey fiber used when at least 3 out of 5 consecutive tests elicited a withdrawal response such as subcutaneous muscle contraction in rats.

[0120] 3 Experimental Results

[0121] In this invention, the pain threshold in all dosage groups was higher than the initial value after 3 days, indicating a longer-lasting analgesic effect. Analysis using GraphPad Prism 9.0 software showed that the 2.5 mg / kg dosage of this invention... -1 5mg·kg -1 10 mg·kg-1 The pain threshold at 72 hours was significantly different from that of the blank incision group (P < 0.0001). Although there was no significant difference (ns) in the pain threshold at 72 hours among the different dosage groups of this invention, the high-dose group had a higher tolerable pain threshold than the low- and medium-dose groups. Furthermore, the pain threshold of the high-dose group was significantly higher than that of the low- and medium-dose groups at 84 hours, indicating a certain dose-dependency. Therefore, the low-, medium-, and high-dose groups of this invention can maintain analgesia for 3 days, demonstrating a long-acting analgesic effect.

[0122] Example 18: Safety Evaluation

[0123] The administration method was the same as above. The healing of the incision at the administration site was observed visually on days 3, 10, and 14 after administration. On day 14, skin samples around the injection site were collected, fixed with 4% paraformaldehyde, embedded in paraffin, and stained with hematoxylin and eosin for histopathological analysis.

[0124] Conclusion: After subcutaneous administration to rats, the incisions of this invention scab over in 3 days, fall off in 10 days, and are basically healed on the back incisions in 14 days, showing no significant difference compared to the untreated group, indicating that this invention has no effect on postoperative incision healing. During days 3, 10, and 14 after administration, the motor behavior of rats in all groups was normal, and no abnormalities were observed at the injection sites and surrounding tissues in all animals. After 14 days, the histopathological changes (hemoptysis, foreign bodies, ulcers) were not significantly different from the untreated group. This invention showed no significant irritation to local tissues, demonstrating good safety and tolerability.

[0125] Example 19: Pharmacokinetic Study in Rats

[0126] 1. In vivo high performance liquid chromatography analysis method

[0127] Chromatographic column: 150mm × 4.6mm octadecylsilane-bonded silica column (5μm)

[0128] Column temperature: 35℃

[0129] Mobile phase: Mobile phase A: Phosphoric acid-methanol-water = 0.1:10:90

[0130] Mobile phase B: Phosphoric acid-water-methanol = 0.1:10:90

[0131] Table 12 Chromatographic conditions

[0132] Flow rate: 1.5 ml / min

[0133] Detection wavelength: 215nm

[0134] Injection volume: 20 μl 2 Plasma sample preparation

[0135] Blank plasma sample: Take blank plasma from rats, thaw it, and accurately measure 100 μL into a 1.5 mL centrifuge tube. Add 700 μL of 4% glacial acetic acid methanol, vortex for 5 min, and centrifuge at 9000 rpm for 10 min to precipitate proteins. Transfer the supernatant to a 1.5 mL centrifuge tube and evaporate to dryness at 40℃. Add 50 μL of phosphoric acid-methanol-water (0.1:10:90) to the residue to reconstitute it, vortex for 6 min, sonicate for 15 min, and centrifuge at 13000 rpm for 20 min. The supernatant is the blank plasma sample solution.

[0136] Plasma samples after drug administration: After thawing, accurately measure 100 μL of rat plasma into a 1.5 mL centrifuge tube, add 10 μL of internal standard (tramadol 2 μg / mL, dissolved in phosphate-methanol-water = 0.1:10:90), and vortex for 1 min to mix thoroughly. Add 700 μL of 4% glacial acetic acid methanol, vortex for 5 min, centrifuge at 9000 rpm for 1 min to precipitate proteins, and transfer the supernatant to a 1.5 mL centrifuge tube. Evaporate to dryness at 40℃. Redissolve the residue in 50 μL of phosphate-methanol-water = 0.1:10:90, vortex for 6 min, sonicate for 15 min to ensure complete drug dissolution, and centrifuge at 13000 rpm for 20 min. The supernatant is the plasma sample solution after drug administration.

[0137] 3. Dosing regimen and sample collection

[0138] Twelve rats were randomly divided into two groups of six each. Each group received an intramuscular injection of the hydrogenated castor oil suspension of this invention (prepared by dissolving the compound of this invention in hydrogenated castor oil) at a dose of 10 mg / kg. Approximately 0.5 mL of blood was collected from the orbital vein of the rats at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, 36, 48, 96, 144, 192, and 240 hours after administration. The blood was added to heparin-treated anticoagulant tubes, centrifuged at 4000 rpm for 10 min, and the supernatant plasma was separated and stored at -20°C for analysis.

[0139] 4. Plasma Sample Detection and Data Processing

[0140] Plasma samples were collected after drug administration and processed according to the method described in Section 2, "Preparation of Plasma Samples." The samples were injected under the chromatographic conditions described in Section 1, "In Vivo High Performance Liquid Chromatography Analysis Method." The peak area ratio of the active pharmaceutical ingredient to the internal standard peak was substituted into the standard curve to calculate the drug concentration. The results were processed using Phoenix software to calculate the relevant pharmacokinetic parameters.

[0141] 5 Results

[0142] Table 13 Results of pharmacokinetic studies in rats

[0143] The results show that the compounds of the present invention can be slowly, continuously and stably released in vivo and converted into tapentaldo, thereby exerting a long-lasting effect. The compounds of the present invention are released stably, achieving a long-lasting release effect.

[0144] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. The compound represented by formula (Ⅰ):

2. The crystal form of the compound according to claim 1, wherein the PXRD pattern of the crystal form has characteristic peaks selected from the following, located at 7.82, 13.56, 15.60, 16.30 and 16.72, wherein the characteristic peaks are represented by 2θ ± 0.2°2θ, CuKα radiation.

3. A process for the preparation of a compound according to claim 1, characterized in that: comprising the step of esterifying heptaminol hydrochloride with sebacoyl chloride in an organic solvent and a base-binding acid agent, the reaction formula being as follows:

4. A process for preparing the crystalline form of claim 2, characterized by: The process includes a crystallization step, wherein the solvent for the crystallization step is selected from one or more of methyl tert-butyl ether, isopropyl ether, n-hexane, and petroleum ether.

5. A process for preparing the crystalline form of claim 4, characterized by: The temperature for the crystallization step is -15 to 0℃.

6. A process for preparing the crystalline form of claim 4, characterized by: This includes pre-filtration using a 0.45μm microporous membrane before the crystallization step, followed by two stages of sterilization filtration using a 0.22μm membrane.

7. A pharmaceutical composition comprising the compound of claim 1 or the crystal form of claim 2, and a pharmaceutically acceptable carrier.

8. The pharmaceutical composition of claim 7, characterized in that: The pharmaceutical composition is intended for administration by injection.

9. Use of the compound of claim 1, the crystal form of claim 2, or the pharmaceutical composition of any one of claims 7-8 in the preparation of a medicament for treating various acute and chronic pains.

10. Use of the compound of claim 1, the crystal form of claim 2, or the pharmaceutical composition of any one of claims 7-8 for the treatment of various acute and chronic pain.

Images