Combined use of NK1 antagonist prodrug compound and 5-HT3 receptor antagonist

Abstract

The present disclosure relates to the combined use of NK1 antagonist prodrug compounds and 5-HT3 receptor antagonists. Specifically, the present disclosure relates to the use of a compound represented by 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.

Description

Technical Field

[0001] The present disclosure belongs to the field of medicine and relates to the use of a rolapitant prodrug in combination with a 5-HT3 receptor antagonist in the preparation of a drug for preventing or treating nausea and / or vomiting. Background Art

[0002] Tachykinins are peptide ligands for neurokinin receptors. Neurokinin receptors, such as NK1, NK2 and NK3, are involved in various biological processes. They can be found in the nervous and circulatory systems of mammals as well as in peripheral tissues. Therefore, modulation of this class of receptors has been studied for the potential treatment or prevention of various diseases in mammals. Typical neurokinin receptor antagonists and their uses include: US5760018 (1998) (pain, inflammation, migraine and vomiting), US5620989 (1997) (pain, nociception and inflammation), WO95 / 19344 (1995), WO 94 / 13639 (1994) and WO 94 / 10165 (1994). 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 a potent, selective NK1 antagonist rolapitant with beneficial therapeutic and pharmacological properties and good metabolic stability, which can be used in the form of a free base or a pharmaceutically acceptable salt, suitable for parenteral administration.

[0004]

[0005] US9101615 provides a prodrug of rolapitant, i.e., a prodrug and a salt thereof in which the hydrogen of the free amine (or two amines) of the compound of formula I is replaced by a group selected from -Y, -X, wherein Y is selected from -P(O)(OH)2, -S(O) n1 R 1 , -C(O)(C 1-6 Alkyl)X, -C(O)(C 1-6 alkyl)(aryl), -C(O)OR 4 ; X is selected from -NR 2 R 3 , -P(O)(OH)2 or -S(O) n1 R1; R 1 Is H or C 1-6 Alkyl; R 2 Is H or C1-6 Alkyl; R 3 Is H or C 1-6 Alkyl; R 4 Is H or C 1-6 Alkyl; n1 is 0-4. The prodrug can be used in a suitable liquid formulation (including or excluding the parenteral delivery vehicle) to treat patients in need thereof.

[0006] On the other hand, drug hemolysis is caused by the massive destruction of red blood cells caused by immune factors after the drug enters the human body. Clinically, anemia, jaundice, soy sauce and urine hemolysis phenomena occur. Drug-induced hemolytic anemia can be divided into the following three types: (1) drug immunity, resulting in antibody-mediated hemolytic reaction; (2) drug action on red blood cells with genetic enzyme defects (such as G6PD deficiency); (3) drug hemolytic reaction to abnormal hemoglobin. The key to treating this disease is to stop using related drugs and control the occurrence of hemolysis to prevent complications. In order to solve the problem of low solubility of the compound of formula I at physiological pH, researchers used a co-solvent-based preparation containing Captisol, propylene glycol and ethanol to significantly improve the solubility of compound 1, but the co-solvent preparation has a significant hemolytic effect after intravenous administration. CN102573475 discloses an improved prescription containing polyethylene glycol 15-hydroxystearate and medium-chain triglycerides. However, the hemolytic effect of the pharmaceutical composition has not been completely solved, even if the compound of formula I is prepared as a prodrug containing phosphate.

[0007] PCT / CN2020 / 098460 provides a new NK1 antagonist prodrug compound that is effective in treating various physiological disorders, symptoms and diseases with fewer side effects, and its structure is shown below:

[0008]

[0009] The present disclosure provides a novel use of a NK1 antagonist prodrug compound which is effective in treating various physiological disorders, symptoms and diseases with less side effects and is combined with a 5-HT3 receptor antagonist in the preparation of a drug for preventing or treating nausea and / or vomiting, and shows good therapeutic effects. Summary of the invention

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

[0011]

[0012] In certain embodiments, the 5-HT3 receptor antagonist is selected from granisetron, ondansetron, ramosetron, tropisetron, palonosetron and dolasetron, preferably palonosetron.

[0013] In certain embodiments, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 10-500 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 The dosage may be once a day, twice a day, three times a day, once a week, or once every two weeks.

[0014] In certain embodiments, the dosage of the 5-HT3 receptor antagonist is 0.075-1 mg, for example, 0.125 mg, 0.25 mg, 0.375 mg, 0.5 mg, 0.625 mg, 0.75 mg, preferably 0.25-0.75 mg. The frequency of administration can be once a day, twice a day, three times a day, once a week, or once every two weeks.

[0015] In certain embodiments, the dosage of the compound of formula (I) or a pharmaceutically 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.

[0016] The pharmaceutically acceptable salt of the drug disclosed herein can be hydrochloride, phosphate, hydrogen phosphate, sulfate, bisulfate, sulfite, acetate, oxalate, malonate, valerate, glutamate, oleate, palmitate, stearate, laurate, borate, p-toluenesulfonate, methanesulfonate, isethionate, maleate, malate, tartrate, benzoate, pamoate, salicylate, vanillate, mandelate, succinate, gluconate, lactobionate or lauryl sulfonate, etc.

[0017] The methods of the present disclosure are used to treat or prevent nausea and / or vomiting caused by a variety of events, including chemotherapy-induced nausea and vomiting ("CINV") caused by moderate or highly emetogenic chemotherapy, radiation-induced nausea and vomiting ("RINV"), and postoperative nausea and vomiting ("PONV"). The methods are preferably implemented shortly before the vomiting-inducing event occurs (i.e., no more than 1 or 2 hours before the event). The methods can be used to treat nausea and / or vomiting during the acute phase or delayed phase of nausea. The combined administration methods of the present disclosure are selected from simultaneous administration, independently formulated and co-administered, or independently formulated and administered sequentially.

[0018] The administration route of the combination disclosed herein is selected from oral administration, parenteral administration, and transdermal administration, and the parenteral administration includes but is not limited to intravenous injection, subcutaneous injection, and intramuscular injection.

[0019] The present disclosure further relates to a method for preventing or treating nausea and / or vomiting, comprising administering to a patient a compound represented by formula (I) or a pharmaceutically acceptable salt thereof and a 5-HT3 receptor antagonist.

[0020] The present disclosure further relates to a method for preventing or treating vomiting, comprising administering a combination of the present disclosure to a patient suffering from vomiting or a patient at risk of vomiting. In other embodiments, the present disclosure provides a method for preventing or treating vomiting by administering one or more combinations described herein. The combination is preferably administered shortly before a vomiting inducing event (i.e., no more than 2 hours before the event). The vomiting may be acute phase vomiting (i.e., vomiting occurring within about 24 hours after a vomiting inducing event), or delayed vomiting (i.e., vomiting occurring after the acute phase, but within 7 days, 6 days, 5 days, or 4 days after a vomiting inducing event). The vomiting may include chemotherapy-induced nausea and vomiting ("CINV"), radiotherapy-induced nausea and vomiting ("RINV"), or postoperative nausea and vomiting ("PONV") caused by moderate or highly emetogenic chemotherapy. In the scheme described in the present disclosure, the combination optionally also includes other components, and the other components include, but are not limited to, other antiemetics, etc.

[0021] The present disclosure also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for preventing or treating nausea and / or vomiting, which is used in combination with a 5-HT3 receptor antagonist.

[0022] The present disclosure also provides a 5-HT3 receptor antagonist for preventing or treating nausea and / or vomiting, which is used in combination with the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0023] The present disclosure also relates to a pharmaceutical composition comprising a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, a 5-HT3 receptor antagonist, and one or more pharmaceutical carriers. The pharmaceutical composition can be prepared into any pharmaceutically acceptable dosage form. For example, it can be formulated into tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injections, sterile powders for injection and concentrated solutions for injection), suppositories, inhalants or sprays.

[0024] The pharmaceutical composition comprising the compound represented by formula (I) or a pharmaceutically acceptable salt thereof and a 5-HT3 receptor antagonist disclosed in the present invention can be administered alone or in combination with one or more therapeutic agents.

[0025] The components to be combined (e.g., the compound represented by formula (I) or its pharmaceutically acceptable salt, 5-HT3 receptor antagonist, and any other component drugs) can be administered simultaneously or separately in sequence. In addition, the components to be combined can also be administered in the form of the same preparation or in the form of separate different preparations.

[0026] The present disclosure also provides a drug packaging box, which contains a pharmaceutical composition of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof and a 5-HT3 receptor antagonist as described in the present disclosure.

[0027] The "combination" described in the present disclosure is a mode of administration, which refers to the administration of at least one dose of the compound shown in formula (I) or its pharmaceutically acceptable salt and at least one dose of chemotherapeutic drug within a certain time limit, wherein the administered drugs all show pharmacological effects. The time limit can be within an administration cycle, preferably within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours, more preferably within 12 hours. The compound shown in formula (I) or its pharmaceutically acceptable salt and the 5-HT3 receptor antagonist can be administered simultaneously or sequentially. This period includes such treatment, in which the compound shown in formula (I) or its pharmaceutically acceptable salt and the 5-HT3 receptor antagonist are administered by the same administration route or different administration routes. The combined administration mode described in the present disclosure is selected from simultaneous administration, independent preparation and co-administration, or independent preparation and sequential administration.

[0028] The term "postoperative nausea and / or vomiting" (PONV) has its conventional meaning in the art. It is well known in the art that it means one or more episodes of vomiting (vomiting and / or retching) or the desire to vomit (nausea) that occur after surgery. Retching involves the same physiological mechanism as vomiting, but occurs on a closed glottis. PONV can be defined as nausea and / or vomiting that occurs within 48 hours after the end of surgery, and can also be defined as nausea and / or vomiting that occurs within 24 hours after the end of surgery.

[0029] "Therapeutically effective amount" refers to the amount of the therapeutic agent that produces the desired effect for which it is applied. In some embodiments, the term refers to an amount sufficient to treat the disease, disorder and / or condition when applied to a population suffering from or susceptible to a disease, disorder and / or condition according to a regimen. 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, disorder and / or condition, and / or delays its onset. Those of ordinary skill in the art will appreciate that the term "therapeutically effective amount" does not actually require successful treatment in a particular individual. On the contrary, a therapeutically effective amount can be an amount that provides a specific desired pharmacological response in a large number of subjects when applied to a patient who needs such treatment. In some embodiments, mentioning a therapeutically effective amount can refer to an amount measured in one or more specific tissues (e.g., tissues affected by a disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those of ordinary skill in the art will appreciate that in some embodiments, a therapeutically effective amount of a specific agent or therapy can be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective agent may be formulated and / or administered in multiple doses, for example, as part of a regimen.

[0030] All numbers herein are to be understood as modified by "about", which, when referring to measurable values ​​such as amounts, durations of time, etc., is meant to encompass variations of ±10%, preferably ±5%, or more. Unless otherwise specified, these variations are appropriate to obtain the desired amount of the drug. Preferably, it is ±1% from the specified value, and even more preferably, ±0.1%.

[0031] The present invention improves the therapeutic effect of diseases such as nausea and / or vomiting by administering the compound represented by formula (I) or a pharmaceutically acceptable salt thereof in combination with a 5-HT3 receptor antagonist. DETAILED DESCRIPTION

[0032] The present disclosure is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the present disclosure.

[0033] The experimental methods in the examples of this disclosure that do not specify specific conditions are usually carried out under conventional conditions or under conditions recommended by raw material or product manufacturers. Reagents that do not specify specific sources are conventional reagents purchased from the market.

[0034] Embodiment 1:

[0035]

[0036] first step:

[0037]

[0038] Under N2 protection, compound 1 (2.43 g, 4.86 mmol, 1 eq) was weighed and dissolved in dichloromethane (36 mL) in a 100 mL three-necked flask, diisopropylethylamine (5 g, 38.76 mmol, 8 eq) was added, cooled to -30 ° C, trimethylsilyl chloride (1.36 g, 12.52 mmol, 2.6 eq) was added, and stirred at room temperature for 2 h. Then 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 and stirred until the reaction was completed. The reaction solution was poured into ice water, separated, extracted with dichloromethane, water and 1N hydrochloric acid solution were added, separated, and then washed with brine, saturated sodium bicarbonate aqueous solution and brine in turn, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 3.0 g of yellow colloid, with a yield of 104%.

[0039] Step 2:

[0040]

[0041] Under N2 protection, compound 2 (2.8 g, 4.53 mmol, 1 eq), tetrabutylammonium iodide (1.68 g, 4.55 mmol, 1 eq), potassium di-tert-butyl phosphate (5.63 g, 22.67 mmol, 5 eq) and dioxane (84 mL) were added to a 500 mL three-necked flask and stirred at 55 °C for 4 h. The reaction solution was cooled, poured into ethyl acetate and water, separated, extracted with ethyl acetate, washed with an aqueous solution of sodium sulfite, then washed with water and brine in turn, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 3.73 g of yellow foam with a yield of 107%.

[0042] Step 3:

[0043]

[0044] Under N2 protection, compound 3 (6.65 g, 8.67 mmol, 1 eq) was added to a 500 mL single-mouth bottle and dissolved in dichloromethane (200 mL). Trifluoroacetic acid (9.89 g, 86.7 mmol, 10.0 eq) was slowly added under ice-cooling, and the reaction was stirred until completion. The mixture was concentrated to obtain 2.29 g of an oily substance, which was purified by a reverse phase silica gel column (C18) (Solution A: 20 mmol NH4HCO3 aqueous solution, Solution B: acetonitrile), and then the pH was adjusted to 1-2 with 1 M phosphoric acid. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 2.7 g of the target product, the compound represented by formula (I).

[0045] Example 2: Toxicity test of the compound represented by formula (I) combined with palonosetron in rats

[0046] Rats were intravenously administered with the compound of formula (I), palonosetron, or the compound of formula (I) + palonosetron once or twice a week for 29 consecutive days. The drug was discontinued for 4 weeks to observe the nature, extent, dose-effect, time-effect relationship and reversibility of possible toxic reactions, determine the toxic target organs or target tissues, and study the toxicokinetics to understand the relationship between exposure dose and toxicological results in toxicity studies.

[0047] Drug preparation

[0048] According to the content conversion, weigh the required amount of the compound represented by formula (I), add an appropriate amount of 0.02M phosphate buffer (pH about 7.40) to prepare to the required concentration. The low-concentration drug preparation can be obtained by diluting the high-concentration drug preparation. After preparation, it is filtered (PES filter membrane) before use. Temporary storage conditions and validity period after preparation: 15-25℃, shaded, use within 72 hours.

[0049] According to the content conversion, weigh the required amount of palonosetron, add an appropriate amount of 0.02M phosphate buffer (pH about 7.40) to prepare to the required concentration. Low-concentration dosing preparations can be obtained by diluting high-concentration dosing preparations. After preparation, filter (PES filter membrane) before use. Temporary storage conditions and validity period after preparation: 15-25℃, shaded, use within 72 hours.

[0050] The required amount of the compound of formula (I) and palonosetron are weighed to the required concentration at a fixed ratio (compound of formula (I): palonosetron free base = 872:1), ensuring that the concentration ratio is compound of formula (I): palonosetron free base = 872:1; the low-concentration drug preparation can be obtained by diluting the high-concentration drug preparation with 0.02M phosphate buffer (pH about 7.40), and after preparation, it is filtered (PES filter membrane) before use. Temporary storage conditions and validity period after preparation: 15-25°C, shaded, used within 72 hours.

[0051] 1.1 Administration

[0052] In this test, the low, medium and high dose groups of the compound represented by formula (I) + palonosetron combination group are designed as (the following doses are expressed as the compound represented by formula (I) + palonosetron) 5+0.0006, 15+0.24, 30+10 mg / kg, respectively. A 5, 20, 40 mg / kg group of the compound represented by formula (I) and a 10 mg / kg group of palonosetron are also set up, and the doses of the compound represented by formula (I) and palonosetron are the same as those of the compound represented by formula (I) and palonosetron in the high dose group of the combination. In addition, a control group is set up, and an equal volume of solvent (0.02M phosphate buffer (pH is about 7.40)) is administered intravenously.

[0053] Intravenous drip, twice a week in the first 4 weeks, once in the 5th week, a total of 9 times, stop the drug and recover for 4 weeks after the last dose, 15 minutes / animal / time, the administration volume is 20ml / kg.

[0054] 1.2 Observe the animal's activities, measure body weight and food intake, test hematology, blood biochemistry, urine routine, ophthalmology, bone marrow smear, and analyze relevant data.

[0055] 1.3 Toxicokinetics

[0056] Sampling time:

[0057] The compound group represented by formula (I), the palonosetron group, and the compound represented by formula (I) + palonosetron dose groups: 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 last administration;

[0058] Control group: before the first and last administration and 2 minutes (±1 minute) after administration;

[0059] Sampling method: Depending on the situation, CO2-O2 mixed gas (volume ratio 7:3) can be used for anesthesia before blood collection;

[0060] Sampling animals: surviving rats from the first and last toxic generation sampling in each group;

[0061] Sampling site: jugular vein;

[0062] Sampling volume: ≥0.3mL;

[0063] Anticoagulant: EDTA-K2;

[0064] Blood sample processing: The whole blood sample is placed in an ice box before centrifugation. Centrifuge at 2-8°C and 1800×g for 10 minutes. Separate the plasma into 2 tubes, one of which is 50μL. The remaining plasma is placed in another tube. After the samples are divided, store them below -66°C for testing.

[0065] 1.4 Gross anatomy and histopathological examination

[0066] 1.5 Statistical analysis

[0067] Quantitative indicators such as body weight, food intake, hematology, blood biochemistry, urine specific gravity, percentage of each cell line and megakaryocyte count (if any) in bone marrow smear examination, organ weight and coefficient were described by mean ± standard deviation (), and qualitative indicators (binary classification, unordered multiple classification and ordered multiple classification) such as urine routine test (except urine specific gravity) and bone marrow smear nucleated cell proliferation (if any) were described by frequency counting. When the sample size was less than 3, the data of this group were not statistically compared.

[0068] The quantitative indicators were first tested for variance homogeneity using the LEVENE test. When the variance was homogeneous (P>0.05), one-way analysis of variance (ANOVA) was used for statistical testing; when the variance was unequal (P≤0.05), the Kruskal-Wallis H rank sum test (KW method) was used for statistical analysis. When the one-way ANOVA showed that the difference was statistically significant (P≤0.05), Dunnett's t test (Dunnett method) was used to compare the differences between the groups; when the one-way ANOVA showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated. When the Kruskal-Wallis H rank sum test showed that the difference was statistically significant (P≤0.05), the Mann-Whitney U test (MW method) was used to compare the differences between the groups; when the Kruskal-Wallis H rank sum test showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated.

[0069] The ordered multi-classification indices were analyzed using the Kruskal-Wallis H rank sum test (KW method); when the differences were statistically significant (P≤0.05), the Mann-Whitney U rank sum test (MW method) was used to compare the differences between the groups.

[0070] The binary indices were analyzed using Fisher's exact probability method (EXACT). When the difference was statistically significant (P ≤ 0.05), Fisher's exact probability method was used to compare the difference between the two groups.

[0071] The inter-group differences were compared between the various dosage groups of the compound represented by formula (I), the palonosetron group, the various dosage groups of the compound represented by formula (I) + palonosetron and the control group.

[0072] All tests were two-sided with α = 0.05. All analyses were performed separately for gender. Body weight, hematology, blood biochemistry, organ weights and coefficients were statistically analyzed using the PRISTIMA version 7.2.0 data acquisition system. The remaining index data were statistically analyzed using Stata / IC 15.0 for Windows.

[0073] Data on general condition observation, ophthalmological examination and pathology were analyzed descriptively.

[0074] Example 3: 29-day toxicity test in rhesus monkeys

[0075] Rhesus monkeys were intravenously administered the compound of formula (I) 1 to 2 times per week for 29 consecutive days, followed by a 4-week recovery period to observe the nature, extent, dose-effect, time-effect relationship and reversibility of possible toxic reactions, determine toxic target organs or target tissues, and study its toxicokinetics to understand the relationship between exposure dose and toxicological results in toxicity studies.

[0076] Drug preparation

[0077] According to the content conversion, weigh the required amount of the compound represented by formula (I), add an appropriate amount of 0.02M phosphate buffer (pH about 7.40) to prepare to the required concentration. The low-concentration drug preparation can be obtained by diluting the high-concentration drug preparation. After preparation, it is filtered (PES filter membrane) before use. Temporary storage conditions and validity period after preparation: 15-25℃, shaded, use within 72 hours.

[0078] 1.1 Administration

[0079] The low, medium and high doses of the compound represented by formula (I) in this test were 2, 10 and 50 mg / kg, respectively. A control group was also set up and given an equal volume of solvent (0.02 M phosphate buffer (pH about 7.40)) in the same manner.

[0080] Intravenous drip, twice a week in the first 4 weeks, once in the 5th week, a total of 9 times, stop the drug and recover for 4 weeks after the last dose, 30 minutes / animal / time, the administration volume is 20ml / kg.

[0081] 1.2 Observe the animal's activities, measure body weight, food intake, body temperature (rectal temperature), electrocardiogram, blood pressure, test hematology, blood biochemistry, urine routine, ophthalmology, bone marrow smear, and analyze relevant data.

[0082] 1.3 Toxicokinetics

[0083] Sampling time:

[0084] 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 and after the first and last administration;

[0085] Control group: 0.25 (±1 min) hours before the first and last administration and after the end of administration;

[0086] Sampled animals: Surviving monkeys from each group;

[0087] Sampling site: saphenous vein of lower limb or other suitable veins;

[0088] Sampling volume: ≥0.4mL;

[0089] Anticoagulant: EDTA-K2;

[0090] Blood sample processing: The whole blood sample is placed in an ice box before centrifugation. Centrifuge at 2-8°C and 1800×g for 10 minutes. Separate the plasma into 2 tubes, one of which is 50μL. The remaining plasma is placed in another tube. After the samples are divided, store them below -66°C for testing.

[0091] 1.4 Gross anatomy and histopathological examination

[0092] 1.5 Statistical analysis

[0093] Quantitative indicators such as body weight, food intake, body temperature, electrocardiogram, blood pressure, hematology, blood biochemistry, urine specific gravity, percentage of each cell line and megakaryocyte count (if any) in bone marrow smear examination, organ weight and coefficient were described by mean ± standard deviation (X ± SD); qualitative indicators (binary classification, unordered multiple classification and ordered multiple classification) such as bone marrow smear nucleated cell proliferation (if any) and urine routine examination (except urine specific gravity) were described by frequency counting. When the sample size was less than 3, the data of this group were not statistically compared.

[0094] The quantitative indicators were first tested for variance homogeneity using the LEVENE test. When the variance was homogeneous (P>0.05), one-way analysis of variance (ANOVA) was used for statistical testing; when the variance was unequal (P≤0.05), the Kruskal-Wallis H rank sum test (KW method) was used for statistical analysis. When the one-way ANOVA showed that the difference was statistically significant (P≤0.05), Dunnett's t test (Dunnett method) was used to compare the differences between the groups; when the one-way ANOVA showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated. When the Kruskal-Wallis H rank sum test showed that the difference was statistically significant (P≤0.05), the Mann-Whitney U test (MW method) was used to compare the differences between the groups; when the Kruskal-Wallis H rank sum test showed that the difference was not statistically significant (P>0.05), the statistical analysis was terminated.

[0095] Ordered multi-classification indicators such as urine examination (clarity, glucose, bilirubin, ketone bodies, occult blood, pH, protein, urobilinogen, and leukocytes) and nucleated cell proliferation in bone marrow smears were analyzed using the Kruskal-Wallis H rank sum test. When the differences were statistically significant (P ≤ 0.05), the Mann-Whitney U test (MW method) was used to compare the differences between the groups.

[0096] The binary (urine nitrite) and unordered multi-classification (urine color) indicators were analyzed using the Fisher exact probability method (EXACT). When the differences were statistically significant (P ≤ 0.05), the Fisher exact probability method was continued to be used to compare the differences between the groups.

[0097] The inter-group differences were compared between the groups of compounds represented by formula (I) and the control group.

[0098] All tests were two-sided with α = 0.05. All analyses were performed separately for gender. Body weight, food intake, hematology, blood biochemistry, organ weights and coefficients were statistically analyzed using the PRISTIMA version 7.2.0 data acquisition system. The remaining indicators were statistically analyzed using Stata / IC 15.0 for Windows.

[0099] Data on general condition observation, ophthalmological examination and pathology were analyzed descriptively.

[0100] Test Example 1: Water Solubility Data and Chemical Stability

[0101] 1.1 Preparation of reagents

[0102] Reagent: NaH2PO4·2H2O

[0103] 1.2 Preparation method

[0104] Prepare according to 100ml specifications as follows:

[0105] pH = 3.0: Phosphate buffer solution: 100 ml 2 mmol / L NaH2PO4, 0.1 M H3PO4 to adjust the pH to 3.0.

[0106] pH = 4.0: Phosphate buffer solution: 100 ml 2 mmol / L NaH2PO4, 0.1 M H3PO4 to adjust the pH to 4.0.

[0107] pH = 7.0: Ultrapure water

[0108] pH = 9.0: Phosphate buffer solution: 100 ml 2mmol / L Na2HPO4, 0.1M NaOH solution to adjust pH to 9.0

[0109] 1.3 Test Method

[0110] Accurately weigh an appropriate amount of the compound to be tested, add it to the solution in small amounts and multiple times, stir and wait for the compound to dissolve, and measure the compound content in the solution. The data are shown in Table 1.

[0111] 2.1 Compound stability test

[0112] Weigh about 1 mg of sample into a vial, place it in a vacuum bag to evacuate, then place it in a container filled with color-changing silica gel, seal it, and prepare two portions in parallel. Prepare enough portions according to the sampling time point and place them at 4 degrees and room temperature respectively. The solubility of the compound represented by formula (I) at different pH values ​​is measured, and the data is shown in Table 1.

[0113] Table 1

[0114] pH Solubility Saturation solubility 7.4 26mg / ml 19.8mg / ml 9.0 28mg / ml 21.4mg / ml

[0115] Note: After 7 days of storage for the best quality, the purity decreases by <0.5%; after 7 days of storage for the medium quality, the purity decreases by 0.5% to 2.0%; after 7 days of storage for the poor quality, the purity decreases by >2.0%

[0116] Test Example 2: Hemolysis

[0117] Red blood cells (RBC) were randomly collected from the rabbit jugular vein or central auricular artery (EDTA whole blood) 10 ml, placed in a glass beaded conical flask and shaken for 10 minutes to remove fibrinogen and make defibrillated blood. About 10 times the amount of sodium chloride injection was added, shaken well, centrifuged at 1500 rpm for 10 minutes, and the supernatant was removed. The precipitated red blood cells were washed 3 times with sodium chloride injection according to the above method until the supernatant was no longer red. The obtained red blood cells were prepared into a 2% (v / v) suspension with sodium chloride injection for later use.

[0118] The test sample (compound represented by formula (I)) was dissolved in PBS (pH 7.4 or pH 5), filtered, and 0.4 mg / ml, 0.8 mg / ml, 1.2 mg / ml, 1.6 mg / ml and 2 mg / ml were prepared for use.

[0119] Take a certain amount of the test sample solution and add the above hemoglobin to the supernatant for testing.

[0120] If the solution in the test tube is clear red, and there are no cells or a small amount of red blood cells remaining at the bottom of the tube, it indicates that hemolysis has occurred; if all the red blood cells sink and the supernatant is colorless and clear, it indicates that no hemolysis has occurred. If there is brown-red or reddish-brown flocculent precipitation in the solution, it does not disperse after gently inverting 3-5 times, indicating that red blood cell aggregation may have occurred; it should be further observed under a microscope, and if red blood cells can be seen to aggregate, it is aggregation. The compounds provided in the present disclosure are measured for hemolysis using this method.

[0121] Conclusion: The compound represented by formula (I) did not show hemolytic effect at a concentration as high as 2 mg / ml.

[0122] Test Example 3: Hemolytic effect of Rolapitant emulsion

[0123] Rolapitant emulsion was prepared according to the method in CN102573475 (prescription: 4.4% polyethylene glycol-15 hydroxystearate, 1.1% medium chain triglyceride and 0.66% soybean oil), and 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 were prepared with PBS for later use.

[0124] The hemolysis was determined according to the method in Test Example 2.

[0125] Conclusion: All concentrations of rolapitant emulsion showed hemolytic effect.

[0126] Test Example 4: Pharmacokinetic Test in Cynomolgus Monkeys

[0127] Using cynomolgus monkeys as test animals, the drug concentration in plasma at different times after injection of the compound represented by formula (I) was determined by LC / MS / MS method to study the pharmacokinetic behavior of the compound in cynomolgus monkeys and evaluate its pharmacokinetic characteristics.

[0128] Drug preparation

[0129] Weigh a certain amount of the compound to be tested and prepare a pH = 4.0 solution with 20 mmol / L sodium dihydrogen phosphate for later use.

[0130] 1.1 Administration

[0131] Intravenous drip, push injection time is about 30 minutes, the dosage is 3.54 mg / kg, the dosage concentration is 2 mg / ml, and the dosage volume is 5 ml / kg.

[0132] 1.2 Operation

[0133] Before administration and 5 minutes, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration, blood was collected from the femoral vein, and about 0.6 mL of each sample was collected, anticoagulated with sodium heparin, and placed on ice immediately after collection. After blood sample collection, it was placed in a labeled centrifuge tube and centrifuged to separate plasma (centrifugation conditions: centrifugal force 2200g, centrifugation 10 minutes, 2-8°C).

[0134] LC / MS / MS was used to determine the contents of the compound represented by formula (I) and rolapitant in plasma samples.

[0135] 1.3 Pharmacokinetic parameter results

[0136] Table 2

[0137]

[0138]

[0139] Note: a) The pharmacokinetic parameters of the compound represented by formula (I) in cynomolgus monkeys, b) The pharmacokinetic parameters of the compound represented by formula (I) metabolized to rolapitant in cynomolgus monkeys.

[0140] Conclusion: In the pharmacokinetic study of the compound represented by formula (I) in cynomolgus monkeys, most of it was rapidly converted into the active metabolite rolapitant in cynomolgus monkeys, which 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 the group consisting of granisetron, ondansetron, ramosetron, tropisetron, palonosetron and dolasetron, 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 nausea and / or vomiting is selected from chemotherapy-induced nausea and vomiting, radiotherapy-induced nausea and vomiting or postoperative nausea and vomiting.

4. The use according to claim 1 or 2, wherein the nausea and / or vomiting is selected from nausea and / or vomiting during the acute phase or the delayed phase of nausea.

5. A pharmaceutical composition comprising a compound of formula (I) as claimed in claim 1 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.

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