Combined use of NK1 antagonist prodrug compounds and 5-HT3 receptor antagonists

Abstract

The present disclosure relates to the combined use of an NK1 antagonist prodrug compound and a 5-HT3 receptor antagonist. Specifically, the present disclosure relates to the combined use of a compound of formula (I) or a pharma- ceutically acceptable salt thereof and a 5-HT3 receptor antagonist in the preparation of a medicament for preventing or treating nausea and / or vomiting. JPEG2024501994000013.jpg63170

Description

[Technical field]

[0001] The present disclosure belongs to the pharmaceutical field and relates to the combined use of a rolapitant prodrug and a 5-HT3 receptor antagonist in the preparation of a medicament for preventing or treating nausea and / or vomiting. [Background technology]

[0002] Tachykinins are peptide ligands for neurokinin receptors. Neurokinin receptors, such as NK1, NK2 and NK3, are involved in a variety of biological processes. They can be found in the nervous and circulatory systems of mammals, and in surrounding tissues. Modulation of such receptors is therefore being investigated to potentially treat or prevent a variety of mammalian disorders. Exemplary neurokinin receptor antagonists and their uses include US 5760018 (1998) (pain, inflammation, migraine and emesis), US 5620989 (1997) (pain, nociception and inflammation), WO 95 / 19344 (1995), WO 94 / 13639 (1994) and WO 94 / 10165 (1994). Further references to other classes of NK1 receptor antagonists include Wu et al., Tetrahedron 56, 3043-3051 (2000), Rombouts et al., Tetrahedron Letters 42, 7397-7399 (2001), and Rogiers et al., Tetrahedron 57, 8971-8981 (2001).

[0003] US7049320 provides an effective and selective NK1 antagonist, rolapitant, having beneficial therapeutic and pharmacological properties and good metabolic stability, which can be applied to formulations for parenteral administration in free base form or in pharma- ceutically acceptable salt form; [ka] .

[0004] US9101615 provides prodrugs of rolapitant, i.e., prodrugs in which the hydrogen of the free amine (or two amines) of the compound of formula I is replaced with a group selected from -Y and -X, where Y is -P(O)(OH)2, -S(O) n1 R 1 , -C(O)(C 1-6 Alkyl group)X, -C(O)(C 1-6 Alkyl group) (aryl group), -C(O)OR 4 X is selected from -NR 2 R 3 , -P(O)(OH)2 or -S(O) n1 Selected from R1, R 1 is H or C 1-6 is an alkyl group, R 2 is H or C 1-6 is an alkyl group, R 3 is H or C 1-6 is an alkyl group, R 4 is H or C 1-6 is an alkyl group, and n1 is 0 to 4. The prodrugs can be used in suitable liquid formulations (with or without the parenteral administration vector) for the treatment of patients in need thereof.

[0005] On the other hand, drug hemolysis is caused by the massive destruction of red blood cells by immune factors after the drug enters the human body, and clinically, hemolysis phenomena such as anemia, jaundice, soy sauce and urine appear. Drug-induced hemolytic anemia can be divided into three types: (1) drug-induced immunity causing antibody-mediated hemolysis, (2) drug action on red blood cells with genetic enzyme deficiency (e.g., G6PD deficiency), and (3) drug hemolysis on abnormal hemoglobin. The key to treating such diseases is to stop the use of related drugs and suppress the occurrence of hemolysis so that complications can be prevented. To solve the problem of the low solubility of compound I at physiological pH, researchers have obviously improved the solubility of compound 1 by using a co-solvent-based formulation containing Captisol, propylene glycol and ethanol, but the co-solvent formulation showed significant hemolysis after intravenous administration. CN102573475 discloses an improved formulation containing polyethylene glycol 15-hydroxystearic acid and medium chain triglyceride. However, even if the compound of formula I is prepared as a prodrug containing phosphate ester, the hemolytic effect of the pharmaceutical composition is still not completely solved.

[0006] PCT / CN2020 / 098460 provides novel NK1 antagonist prodrug compounds that are effective for treating various physiological disorders, symptoms and diseases with minimal side effects, and the structures thereof are as follows: [ka] .

[0007] The present disclosure provides a combination use of a novel NK1 antagonist prodrug compound and a 5-HT3 receptor antagonist in the preparation of a medicament for preventing or treating nausea and / or vomiting, which is effective for treating various physiological disorders, symptoms and diseases with minimal side effects, and shows good therapeutic effects. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] US5760018 [Patent Document 2] US5620989 [Patent Document 3] WO95 / 19344 [Patent Document 4] WO 94 / 13639 [Patent Document 5] WO 94 / 10165 [Patent Document 6] US7049320 [Patent Document 7] US9101615 [Patent Document 8] CN102573475 [Patent Document 9] PCT / CN2020 / 098460 [Non-patent literature]

[0009] [Non-Patent Document 1] Wu et al., Tetrahedron 56, 3043~3051(2000) [Non-Patent Document 2] Rombouts et al., Tetrahedron Letters 42, 7397~7399 (2001) [Non-Patent Document 3] Rogiers et al., Tetrahedron 57, 8971~8981 (2001) Summary of the Invention [Means for solving the problem]

[0010] The present disclosure provides the use of a compound of formula (I) or a pharma- ceutically acceptable salt thereof in combination with a 5-HT3 receptor antagonist in the preparation of a medicament for preventing or treating nausea and / or vomiting, [ka] .

[0011] In some embodiments, the 5-HT3 receptor antagonist is selected from granisetron, ondansetron, ramosetron, tropisetron, palonosetron and dolasetron, preferably palonosetron.

[0012] In some embodiments, the dosage of the compound represented by formula (I) or a pharma- ceutically acceptable salt thereof is 10 to 500 mg, for example, 10 mg, 20 mg, 27.25 mg, 30 mg, 40 mg, 50 mg, 54.5 mg, 60 mg, 70 mg, 80 mg, 81.75 mg, 90 mg, 100 mg, 109 mg, 110 mg, 120 mg, 130 mg, 136.25 mg, 140 mg, 150 mg, 160 mg, 163.5 mg, 170 mg, 180 mg, 190 mg, 190.75 mg, 200 mg, 210 mg, 218 mg, 220 mg, 230 mg, 240 mg, 245.25 mg, 250 mg, 260 mg, 270 mg, 272.5 mg, 280 mg, 290 mg, 299.75 mg, mg, 300 mg, 310 mg, 320 mg, 327 mg, 330 mg, 340 mg, 350 mg, 354.25 mg, 360 mg, 370 mg, 380 mg, 381.5 mg, 390 mg, 400 mg, 408.75 mg, 410 mg, 420 mg, 430 mg, 436 mg, 440 mg, 450 mg, 460 mg, 463.25 mg, 470 mg, 480 mg, 490 mg, 490.5 mg, 500 mg. The dosing frequency may be once daily, twice daily, three times daily, once weekly, or once every two weeks.

[0013] In some embodiments, the dose of the 5-HT3 receptor antagonist is 0.075 to 1 mg, for example, 0.125 mg, 0.25 mg, 0.375 mg, 0.5 mg, 0.625 mg, or 0.75 mg, and preferably 0.25 to 0.75 mg. The frequency of administration may be once a day, twice a day, three times a day, once a week, or once every two weeks.

[0014] In some embodiments, the dosage of the compound represented by formula (I) or a pharma- ceutically acceptable salt thereof is 27.25 mg, 54.5 mg, 81.75 mg, 109 mg, 136.25 mg, 163.5 mg, 190.75 mg, 218 mg, 245.25 mg, 272.5 mg, 299.75 mg, 327 mg, 354.25 mg, 381.5 mg, 408.75 mg, 436 mg, 463.25 mg, or 490.5 mg, and the administration frequency may be once a day, twice a day, three times a day, once a week, or once every two weeks. The dosage of the 5-HT3 receptor antagonist is 0.125 mg, 0.25 mg, 0.375 mg, 0.5 mg, 0.625 mg or 0.75 mg, and the administration frequency may be once a day, twice a day, three times a day, once a week or once every two weeks.

[0015] Pharmaceutically acceptable salts of the agents described in the present disclosure may be hydrochloride, phosphate, hydrogen phosphate, sulfate, hydrogen sulfate, sulfite, acetate, oxalate, malonate, pentanoate, glutamate, oleate, palmitate, stearate, laurate, borate, p-toluenesulfonate, methanesulfonate, isethionate, maleate, malate, tartrate, benzoate, pamoate, salicylate, vanillate, mandelate, succinate, gluconate, lactobionate or laurylsulfonate, and the like.

[0016] The disclosed methods are used to treat or prevent nausea and / or vomiting due to a variety of events, including chemotherapy-induced nausea and vomiting ("CINV") due to moderately or highly emetogenic chemotherapy, radiation therapy-induced nausea and vomiting ("RINV"), and postoperative nausea and vomiting ("PONV"). The methods are preferably performed immediately prior to the occurrence of an emetogenic event (i.e., within 1 or 2 hours before the event). The methods can be used to treat acute or delayed nausea and / or vomiting. The combination administration methods described herein are selected from simultaneous administration, separate preparations and co-administration, or separate preparations and sequential administration.

[0017] The combined administration routes described in the present disclosure are selected from oral administration, parenteral administration, and transdermal administration, where parenteral administration includes, but is not limited to, intravenous injection, subcutaneous injection, and intramuscular injection.

[0018] The present disclosure further relates to a method of preventing or treating nausea and / or vomiting comprising administering to a patient a compound of formula (I) or a pharma- ceutically acceptable salt thereof and a 5-HT3 receptor antagonist.

[0019] The present disclosure further relates to a method of preventing or treating emesis comprising administering a combination of the present disclosure to a patient suffering from or at risk of suffering from emesis. In another embodiment, the present disclosure provides a method of preventing or treating emesis by administering one or more combinations described herein. The combination is preferably administered immediately prior to the occurrence of an emesis-inducing event (i.e., within 2 hours before the event). The emesis may be acute emesis (i.e., emesis occurring within about 24 hours after the emesis-inducing event), or delayed emesis (i.e., emesis occurring after the acute phase, but within 7, 6, 5, or 4 days after the emesis-inducing event). The emesis may include chemotherapy-induced nausea and vomiting ("CINV") due to moderately or highly emetogenic chemotherapy, radiation therapy-induced nausea and vomiting ("RINV"), or postoperative nausea and vomiting ("PONV"). In the forms described herein, the combinations optionally further include other ingredients, including, but not limited to, other antiemetics.

[0020] The disclosure further provides a compound of formula (I) or a pharma- ceutically acceptable salt thereof, in combination with a 5-HT3 receptor antagonist, for preventing or treating nausea and / or vomiting.

[0021] The present disclosure further provides a 5-HT3 receptor antagonist in combination with a compound of formula (I) or a pharma- ceutically acceptable salt thereof for preventing or treating nausea and / or vomiting.

[0022] The present disclosure further relates to a pharmaceutical composition comprising a compound of formula (I) or its pharma- ceutical acceptable salt, a 5-HT3 receptor antagonist, and one or more pharma- ceutical acceptable vectors.The pharmaceutical composition can be prepared into any one of pharma- ceutical acceptable dosage forms.For example, it can be prepared into tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injection solutions, sterile powders for injections, and concentrated solutions for injections), suppositories, inhalants, or sprays.

[0023] The pharmaceutical compositions described herein, including the compound of formula (I) or a pharma- ceutical acceptable salt thereof, a 5-HT3 receptor antagonist, may be administered alone or in combination with one or more therapeutic agents.

[0024] Each component to be combined (e.g., the compound represented by formula (I) or a pharma- ceutically acceptable salt thereof, the 5-HT3 receptor antagonist, and any other component drug) may be administered simultaneously or separately in sequence. Furthermore, each component to be combined may be administered in the same formulation or in separate formulations.

[0025] The present disclosure further provides a pharmaceutical package containing a pharmaceutical composition of a compound of formula (I) or a pharma- ceutically acceptable salt thereof, as described herein, and a 5-HT3 receptor antagonist.

[0026] The "combination" described in the present disclosure is a method of administration in which at least one dose of a compound represented by formula (I) or a pharma- ceutically acceptable salt thereof and at least one dose of a chemotherapeutic agent are administered within a certain period of time, and both of the administered drugs exhibit pharmacological action. The above period may be within one administration cycle, and is preferably within 4 weeks, 3 weeks, 2 weeks, 1 week, or within 24 hours, and more preferably within 12 hours. The compound represented by formula (I) or a pharma- ceutically acceptable salt thereof and the 5-HT3 receptor antagonist can be administered simultaneously or sequentially. Such a period includes treatments in which the compound represented by formula (I) or a pharma- ceutically acceptable salt thereof and the 5-HT3 receptor antagonist are administered by the same or different administration routes. The combination administration method described in the present disclosure is selected from simultaneous administration, co-administration after separate preparation, or successive administration after separate preparation.

[0027] The term "postoperative nausea and / or vomiting" (PONV) adopts its common meaning in the art. As is well known in the art, it refers to the occurrence of one or several emetic attacks (vomiting and / or retching) or the urge to vomit (nausea) following surgery. Nausea involves similar physiological mechanisms as vomiting, but occurs with the vocal cords closed. PONV may be defined as nausea and / or vomiting occurring within 48 hours after the completion of surgery, or nausea and / or vomiting occurring within 24 hours after the completion of surgery.

[0028] A "therapeutically effective amount" refers to an amount of a therapeutic agent whose administration produces a desired effect. In some embodiments, the term refers to an amount sufficient to treat a disease, disease, and / or condition when administered to a population suffering from or susceptible to said disease, disease, and / or condition by way of form. In some embodiments, a therapeutically effective amount is an amount that reduces the incidence and / or severity of one or more symptoms of a disease, disease, and / or condition and / or delays its onset. Those skilled in the art will appreciate that the term "therapeutically effective amount" does not necessarily result in successful treatment in a particular individual. Conversely, a therapeutically effective amount may be an amount that provides a particular desired pharmacological response in a number of subjects when administered to a patient in need of such treatment. In some embodiments, reference to a therapeutically effective amount can refer to, for example, an amount measured in one or more particular tissues (e.g., tissues affected by a disease, disease, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those of skill in the art will appreciate that in some embodiments, a therapeutically effective amount of a particular reagent or treatment can be prepared and / or administered in a single dose. In some embodiments, a therapeutically effective agent can be prepared and / or administered in multiple doses, e.g., as part of a dosage form.

[0029] All numbers in this specification may be understood to be modified by "about", e.g., when referring to measurable values ​​such as amounts or time durations, and are meant to cover variations of ±10%, preferably ±5%, or more. Unless otherwise stated, these variations are appropriate to obtain the required amount of drug. Preferably, the variations are ±1%, and even more preferably ±0.1% from the specified value.

[0030] The present disclosure provides improved therapeutic effects for disorders such as nausea and / or vomiting by co-administering a compound represented by formula (I) or a pharma- ceutically acceptable salt thereof with a 5-HT3 receptor antagonist. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] The present disclosure will be further described below in conjunction with examples, but these examples are not intended to limit the scope of the present disclosure.

[0032] Experimental methods for which specific conditions are not specified in the examples of this disclosure generally follow conventional conditions or conditions recommended by the raw material or commercial manufacturer. Reagents for which a specific source is not specified are conventional reagents that are commercially available.

[0033] Example 1: [ka]

[0034] Step 1: [ka] Under N2 protection, compound 1 (2.43 g, 4.86 mmol, 1 eq) was weighed into a 100 mL three-neck flask and dissolved in dichloromethane (36 mL). Diisopropylethylamine (5 g, 38.76 mmol, 8 eq) was added, the mixture was cooled to -30°C, trimethylchlorosilane (1.36 g, 12.52 mmol, 2.6 eq) was added, and the mixture was stirred at room temperature for 2 h. The mixture was further cooled to -25°C, a dichloromethane solution of chloromethyl chloroformate (0.77 g, 6 mmol, 1.23 eq) was added dropwise, and the temperature was controlled at -20°C to -5°C while stirring until the reaction was completed. The reaction solution was poured into ice water, separated, extracted with dichloromethane, water and 1 N hydrochloric acid solution were added, separated, washed successively with saline, a saturated aqueous solution of sodium bicarbonate and saline, dried over anhydrous sodium sulfate, filtered and concentrated to obtain 3.0 g of a yellow gel-like product with a yield of 104%.

[0035] Step 2: [ka] Compound 2 (2.8 g, 4.53 mmol, 1 eq), tetrabutylammonium iodide (1.68 g, 4.55 mmol, 1 eq), di-tert-butyl potassium phosphate (5.63 g, 22.67 mmol, 5 eq) and dioxane (84 mL) were added to a 500 mL three-neck flask under N2 protection, heated to 55°C and stirred for 4 h. The reaction mixture was cooled, poured into ethyl acetate and water, separated, extracted with ethyl acetate, washed with an aqueous solution of sodium sulfite, water and brine, dried over anhydrous sodium sulfate, filtered and concentrated to give 3.73 g of a yellow foam in 107% yield.

[0036] Step 3: [ka] Compound 3 (6.65 g, 8.67 mmol, 1 eq) was dissolved in dichloromethane (200 mL) in a 500 mL single-neck flask under N2 protection, trifluoroacetic acid (9.89 g, 86.7 mmol, 10.0 eq) was slowly added while cooling with ice water, and the mixture was stirred until the reaction was completed. The mixture was concentrated to obtain 2.29 g of an oily product, which was purified by reverse phase silica gel column (C18) purification (Solution A: 20 mmol of NH4HCO3 aqueous solution, Solution B: acetonitrile), and further adjusted to pH=1-2 with 1 M phosphoric acid, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 2.7 g of the desired product, compound of formula (I).

[0037] Example 2: Toxicity study of the combination of a compound of formula (I) with palonosetron in rats Rats were administered the compound of formula (I), palonosetron or the compound of formula (I) + palonosetron combination by intravenous injection once or twice a week for 29 days, and then allowed to recover for 4 weeks after discontinuation to observe the characteristics, degree, dose-effect and time-effect relationship and reversibility of possible toxic reactions, determine the toxic target organs or tissues, and study their toxicokinetic properties, and clarify the relationship between the exposure dose and toxicological results in the toxicity study.

[0038] Drug preparation The required amount of the compound represented by formula (I) was weighed out based on the content, and an appropriate amount of 0.02 M phosphate buffer (pH about 7.40) was added to prepare the desired concentration. The low-concentration formulation was obtained by diluting the high-concentration formulation, and after preparation, it was filtered (PES filtration membrane) before use. Temporary storage conditions and expiration date after preparation: 15-25℃, in a dark place, use within 72 hours.

[0039] The required amount of palonosetron was weighed out based on the content, and an appropriate amount of 0.02 M phosphate buffer (pH approximately 7.40) was added to adjust the concentration to the desired level. The low-concentration formulation was obtained by diluting the high-concentration formulation, and after preparation, it was filtered (through a PES filtration membrane) before use. Temporary storage conditions and expiration date after preparation: 15-25°C, in a dark place, use within 72 hours.

[0040] The required amount of the compound represented by formula (I) and palonosetron were weighed out in a certain ratio (compound represented by formula (I):palonosetron free base=872:1) until the desired concentration was reached, and the concentration ratio was ensured to be compound represented by formula (I):palonosetron free base=872:1. The low-concentration formulation was obtained by diluting the high-concentration formulation with 0.02 M phosphate buffer (pH about 7.40), and was used after filtration (PES filtration membrane) after preparation. Temporary storage conditions and expiration date after preparation: 15-25°C, in a dark place, use within 72 hours.

[0041] 1.1 Administration The low-dose group, the medium-dose group, and the high-dose group of the compound of formula (I) + palonosetron combination group in this study were respectively designed as 5 + 0.0006, 15 + 0.24, and 30 + 10 mg / kg (the following doses are shown for the compound of formula (I) + palonosetron). In addition, 5, 20, and 40 mg / kg groups of the compound of formula (I) and 10 mg / kg group of palonosetron were set up, and the doses of the compound of formula (I) and the palonosetron were the same as those of the compound of formula (I) and the palonosetron in the combination high-dose group. In addition, a control group was set up, and an equal volume of solvent (0.02 M phosphate buffer (pH about 7.40)) was administered by intravenous injection.

[0042] The mice were administered intravenously twice a week for the first 4 weeks and once a week in the fifth week, for a total of nine doses. After the final dose, the mice were allowed to recover for four weeks and then discontinued. The administration was performed over 15 minutes per mouse at a volume of 20 mL / kg.

[0043] 1.2 The animals' activity status was observed, their weight and food intake were measured, and the hematology, blood biochemistry, routine urinalysis, ophthalmology, and bone marrow smear samples were detected and examined to analyze the relevant data.

[0044] 1.3 Toxicokinetics Sampling Time: The compound of formula (I), palonosetron, and each dose group of the compound of formula (I) + palonosetron: 2 minutes (± 30 seconds), 0.25 (± 1 minute), 0.5 hours (± 1 minute), 2 hours (± 1 minute), 4 hours (± 2 minutes), 8 hours (± 5 minutes), 24 hours (± 30 minutes), and 48 hours (± 30 minutes) before and after the first and final administration. Control group: 2 minutes (± 1 minute) before and after the first and final administration. Sampling method: In some cases, subjects can be anesthetized with a CO2-O2 gas mixture (volume ratio 7:3) before blood collection. Animals sampled: Rats surviving the first and last toxicological sampling of each group. Sampling site: jugular vein. Sampling volume: ≥ 0.3 mL. Anticoagulant: EDTA-K2. Blood sample processing: The whole blood sample was placed in an ice box before centrifugation, and centrifuged at 2-8℃ and 1800×g for 10 min. The plasma was separated into two tubes, with 50 μL in one tube and the remaining plasma in the other tube. After the sample was dispensed, it was stored at -66℃ or below for detection.

[0045] 1.4 Gross Anatomy and Histopathology

[0046] 1.5 Statistical analysis Quantitative indices such as body weight, food intake, hematology, blood biochemistry, urine specific gravity, bone marrow smears for each cell line percentage and megakaryocyte count (if present), organ weights and coefficients were described as mean numbers ± standard deviation ( ), while qualitative indices (binary classification, disordered multiclass classification and ordered multiclass classification) such as routine urinalysis (except urine specific gravity), nucleated cell proliferation (if present) in bone marrow smears were described as frequency counts. When the number of samples was less than 3, the data of the groups were not statistically compared.

[0047] Quantitative indices were first tested for equality of variance using the Levene test, and if the variances were equal (P>0.05), statistical testing was performed using one-way analysis of variance (ANOVA), and if the variances were unequal (P≦0.05), statistical analysis was performed using the Kruskal-Wallis H rank sum test (KW method). If the one-way ANOVA showed that the differences were statistically significant (P≦0.05), the differences between groups were compared using Dunnett's t test (Dunnett method), and if the one-way ANOVA showed that the differences were not statistically significant (P>0.05), statistical analysis was terminated. If the Kruskal-Wallis H rank sum test showed that the differences were statistically significant (P≦0.05), the Mann-Whitney U test (MW method) was used to compare the differences between groups, and if the Kruskal-Wallis H rank sum test showed that the differences were not statistically significant (P>0.05), the statistical analysis was terminated.

[0048] Ordinal multiclass indices were analyzed by the Kruskal-Wallis H rank sum test (KW method), and if the differences were statistically significant (P ≤ 0.05), the differences between groups were compared by the Mann-Whitney U rank sum test (MW method).

[0049] Binary classification indices were analyzed by Fisher's exact probability test (EXACT), and if the differences were statistically significant (P≦0.05), the differences between the two groups were subsequently compared by Fisher's exact probability test.

[0050] Comparison of differences between groups was performed between each dose group of the compound of formula (I), the palonosetron group, each dose group of the compound of formula (I) + palonosetron, and the control group.

[0051] All tests were two-sided with α = 0.05. All analyses were performed by sex. Body weight, hematology, blood biochemistry, organ weights and coefficients were statistically analyzed using the PRISTIMA 7.2.0 data collection system. Other indicator data were statistically analyzed using Stata / IC 15.0 for Windows.

[0052] Data from general observations, ophthalmological examinations, and pathological examinations were analyzed descriptively.

[0053] Example 3: 29-day toxicity study in rhesus monkeys The compound of formula (I) was administered intravenously to rhesus monkeys once or twice a week for 29 days, followed by a 4-week recovery period to observe the characteristics, severity, dose-effect and time-effect relationship, and reversibility of potential toxic reactions, to determine the toxic target organs or tissues, and to study its toxicokinetic properties, and to clarify the relationship between the exposure dose and the toxicological results in the toxicity study.

[0054] Drug preparation The required amount of the compound represented by formula (I) was weighed out based on the content, and an appropriate amount of 0.02 M phosphate buffer (pH about 7.40) was added to prepare the desired concentration. The low-concentration formulation was obtained by diluting the high-concentration formulation, and after preparation, it was filtered (PES filtration membrane) before use. Temporary storage conditions and expiration date after preparation: 15-25℃, in a dark place, use within 72 hours.

[0055] 1.1 Administration The low, medium and high doses of the compound of formula (I) in this test were 2, 10 and 50 mg / kg, respectively. A control group was also administered an equal volume of the solvent (0.02 M phosphate buffer (pH about 7.40)).

[0056] The mice were administered intravenously twice a week for the first 4 weeks and once a week in the fifth week, for a total of nine doses. After the final dose, the mice were allowed to recover for four weeks and then discontinued. The administration was administered intravenously for 30 minutes per mouse at a volume of 20 mL / kg.

[0057] 1.2 The animals' activity status was observed, and their weight, food intake, body temperature (rectal temperature), electrocardiogram, and blood pressure were measured, and hematology, blood biochemistry, routine urinalysis, ophthalmology, and bone marrow smears were detected and examined to analyze the relevant data.

[0058] 1.3 Toxicokinetics Sampling Time: For each dose group of the compound represented by formula (I): 2 minutes (±30 seconds), 0.25 (±1 minute), 0.5 (±1 minute), 2 (±2 minutes), 4 (±2 minutes), 8 (±2 minutes), 24 (±5 minutes), and 48 hours (±1 minute) before the first and final administration and after the end of administration. Control group: 0.25 (± 1 min) before and after the first and last administration. Animals sampled: Surviving monkeys from each group. Sampling site: saphenous vein or other suitable vein. Sampling volume: ≥ 0.4 mL. Anticoagulant: EDTA-K2. Blood sample processing: The whole blood sample was placed in an ice box before centrifugation, and centrifuged at 2-8℃ and 1800×g for 10 min. The plasma was separated into two tubes, with 50 μL in one tube and the remaining plasma in the other tube. After the sample was dispensed, it was stored at -66℃ or below for detection.

[0059] 1.4 Gross Anatomy and Histopathology

[0060] 1.5 Statistical analysis Quantitative indices such as body weight, food intake, body temperature, electrocardiogram, blood pressure, hematology, blood biochemistry, urine specific gravity, bone marrow smears for the percentage of each cell line and the number of megakaryocytes (if available), and organ weights and coefficients were calculated as mean values ​​± standard deviation (

number

[0061] Quantitative indices were first tested for equality of variance using the Levene test, and if the variances were equal (P>0.05), statistical testing was performed using one-way analysis of variance (ANOVA), and if the variances were unequal (P≦0.05), statistical analysis was performed using the Kruskal-Wallis H rank sum test (KW method). If the one-way ANOVA showed that the differences were statistically significant (P≦0.05), the differences between groups were compared using Dunnett's t test (Dunnett method), and if the one-way ANOVA showed that the differences were not statistically significant (P>0.05), statistical analysis was terminated. If the Kruskal-Wallis H rank sum test showed that the differences were statistically significant (P≦0.05), the Mann-Whitney U test (MW method) was used to compare the differences between groups, and if the Kruskal-Wallis H rank sum test showed that the differences were not statistically significant (P>0.05), the statistical analysis was terminated.

[0062] For example, ordinal multiclass indicators such as urinalysis (clarity, glucose, bilirubin, ketone bodies, occult blood, pH value, protein, urobilinogen, white blood cells) and nucleated cell proliferation status in bone marrow smears were analyzed by the Kruskal-Wallis H rank sum test (KW method), and if the differences were statistically significant (P ≤ 0.05), the differences between groups were compared by the Mann-Whitney U rank sum test (MW method).

[0063] Bicategory (urinary nitrite) and disordered multicategory (urine color) indices were analyzed by Fisher's exact probability test (EXACT), and if differences were statistically significant (P ≤ 0.05), differences between both groups were subsequently compared by Fisher's exact probability test.

[0064] Comparison of differences between groups was performed between each group of the compound of formula (I) and the control group.

[0065] All tests were two-sided with α = 0.05. All analyses were performed by sex. Body weight, food intake, hematology, blood biochemistry, organ weights and coefficients were statistically analyzed using the PRISTIMA 7.2.0 data collection system. Other indices were statistically analyzed using Stata / IC 15.0 for Windows.

[0066] Data from general observations, ophthalmological examinations, and pathological examinations were analyzed descriptively.

[0067] Test Example 1: Water Solubility Data and Chemical Stability 1.1 Drug preparation Reagent: NaH2PO4·2H2O 1.2 Preparation method Prepare the following for a 100 mL batch: pH=3.0: Phosphate buffer solution: 100 mL of 2 mmol / L NaH2PO4, adjust pH to 3.0 with 0.1 M H3PO4.

[0068] pH=4.0: Phosphate buffer solution: 100 mL of 2 mmol / L NaH2PO4, adjust pH to 4.0 with 0.1 M H3PO4.

[0069] pH=7.0: Ultrapure water pH=9.0: Phosphate buffer solution: 100 mL of 2 mmol / L Na2HPO4, adjust pH to 9.0 with 0.1 M NaOH solution 1.3 Test Method An appropriate amount of the compound to be detected was precisely weighed, and the solution was added in small portions and stirred until the compound was dissolved, and the content of the compound in the solution was measured. The data is shown in Table 1.

[0070] 2.1 Compound stability experiments Approximately 1 mg of sample was weighed into a vial, placed in a vacuum bag and evacuated, then placed in a container containing discoloured silica gel, sealed and prepared in parallel in two portions. Sufficient portions were prepared depending on the sampling time point and kept at 4°C and room temperature, respectively. The solubility of compound of formula (I) at different pH values ​​was measured and the data are shown in Table 1.

[0071] [Table 1]

[0072] Remarks: Good: After 7 days, the purity loss is <0.5%; Moderate: After 7 days, the purity loss is 0.5%~2.0%; Poor: After 7 days, the purity loss is >2.0%

[0073] Test Example 2: Hemolytic effect 10 mL of red blood cells (RBCs) were randomly collected from the jugular vein or central ear artery of rabbits (EDTA whole blood), placed in an Erlenmeyer flask with a glass stopper, and shaken for 10 minutes to remove fibrinogen and prepare defibrinated blood. Approximately 10 times the volume of sodium chloride injection was added, shaken uniformly, and centrifuged at 1500 rpm for 10 minutes, the supernatant was removed, and the precipitated red blood cells were washed three times with sodium chloride injection according to the above method until the supernatant was no longer red. The obtained red blood cells were suspended in sodium chloride injection at 2% (v / v) and prepared for use.

[0074] The test sample (compound represented by formula (I)) was dissolved in PBS (pH 7.4 or pH 5), filtered, and adjusted to 0.4 mg / mL, 0.8 mg / mL, 1.2 mg / mL, 1.6 mg / mL, and 2 mg / mL for use.

[0075] An aliquot of the test sample solution was tested in the supernatant with the above hemoglobin.

[0076] If the solution in the test tube is red and transparent, and no cells remain at the bottom of the tube, or a few red blood cells remain, this indicates that hemolysis has occurred, and if all the red blood cells have settled and the supernatant is colorless and transparent, this indicates that hemolysis has not occurred. If there is a brown or red-brown flocculent precipitate in the solution that does not disperse even after gently inverting it 3 to 5 times, this indicates that red blood cell aggregation may have occurred, and it is necessary to further observe under a microscope. If a cluster of red blood cells is observed, this is considered to be aggregation. The hemolytic activity of the compound provided by the present disclosure was measured by this method.

[0077] Conclusion: The compound of formula (I) did not exhibit hemolytic activity even at a concentration as high as 2 mg / mL.

[0078] Test Example 3: Hemolytic effect of rolapitant emulsion Rolapitant emulsions (formulation: 4.4% polyethylene glycol 15-hydroxystearate, 1.1% medium chain triglyceride and 0.66% soybean oil) were prepared with reference to the method in CN102573475, and diluted with PBS to 0.18 mg / mL, 0.09 mg / mL, 0.045 mg / mL, 0.023 mg / mL, 0.011 mg / mL, 0.056 mg / mL and 0.028 mg / mL for use.

[0079] The hemolytic activity was measured with reference to the method in Test Example 2.

[0080] Conclusion: All concentrations of rolapitant emulsion have hemolytic effect.

[0081] Test Example 4: Pharmacokinetic study in cynomolgus monkeys Cynomolgus monkeys were used as test animals, and the plasma drug concentrations of the compound of formula (I) were measured at different time points after injection by LC / MS / MS. The pharmacokinetic behavior of the compound in the cynomolgus monkeys was studied, and its pharmacokinetic characteristics were evaluated.

[0082] Drug preparation A fixed amount of the compound to be detected was weighed out and prepared into a solution of pH=4.0 with 20 mmol / L sodium dihydrogen phosphate for use.

[0083] 1.1 Administration The drug was administered by intravenous drip infusion for approximately 30 minutes at a dose of 3.54 mg / kg, a concentration of 2 mg / mL, and a volume of 5 mL / kg.

[0084] 1.2 Operation Blood samples were taken from the femoral vein before administration and 5 min, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 24 h after administration. Approximately 0.6 mL of each sample was collected, anticoagulated with sodium heparin, and placed on ice immediately after collection. After blood samples were collected, they were placed in labeled centrifuge tubes and centrifuged to separate the plasma (centrifugation conditions: centrifugal force 2200 g, 10 min centrifugation, 2-8°C).

[0085] The contents of the compound of formula (I) and rolapitant in the plasma samples were determined by LC / MS / MS.

[0086] 1.3 Pharmacokinetic parameter results [Table 2]

[0087] Note: a is the pharmacokinetic parameter of the compound of formula (I) in cynomolgus monkeys, and b is the pharmacokinetic parameter of the compound of formula (I) metabolized to rolapitant in cynomolgus monkeys.

[0088] Conclusion: The compound of formula (I) is almost rapidly converted to the active metabolite rolapitant in cynomolgus monkeys in pharmacokinetic studies and has good pharmacokinetic properties.

Claims

1. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a 5-HT3 receptor antagonist in the preparation of a medicament for preventing or treating nausea and / or vomiting, wherein the 5-HT3 receptor antagonist is selected from granisetron, ondansetron, ramosetron, tropisetron, palonosetron and dolasetron. 【Chemistry 1】 。

2. The use according to claim 1, wherein the 5-HT3 receptor antagonist is palonosetron.

3. The use according to claim 1 or 2, wherein the dose of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is 10 to 500 mg.

4. The use according to any one of claims 1 to 3, wherein the dosage of the 5-HT3 receptor antagonist is 0.075 to 1 mg.

5. The use according to any one of claims 1 to 4, wherein the administration frequency of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from once a day, twice a day, three times a day, once a week, and once every two weeks.

6. The use according to any one of claims 1 to 5, wherein the administration frequency of the 5-HT3 receptor antagonist is selected from once a day, twice a day, three times a day, once a week, and once every two weeks.

7. The use according to any one of claims 1 to 6, wherein the nausea and / or vomiting is selected from chemotherapy-induced nausea and vomiting, radiotherapy-induced nausea and vomiting, or post-operative nausea and vomiting.

8. The use according to any one of claims 1 to 7, wherein the nausea and / or vomiting is selected from acute nausea and / or delayed nausea and / or vomiting.

9. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, a 5-HT3 receptor antagonist, and one or more pharmaceutically acceptable carriers, wherein the 5-HT3 receptor antagonist is selected from granisetron, ondansetron, ramosetron, tropisetron, palonosetron, and dolasetron.

10. A pharmaceutical packaging box containing a pharmaceutical composition of a compound represented by formula (I) or a pharmaceutically acceptable salt thereof and a 5-HT3 receptor antagonist, wherein the 5-HT3 receptor antagonist is selected from granisetron, ondansetron, ramosetron, tropisetron, palonosetron, and dolasetron.

11. A pharmaceutical composition for preventing or treating nausea and / or vomiting, comprising a compound represented by formula (I) or a pharmaceutically acceptable salt thereof and a 5-HT3 receptor antagonist, wherein the 5-HT3 receptor antagonist is selected from granisetron, ondansetron, ramosetron, tropisetron, palonosetron, and dolasetron.

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