A kind of amide compound, preparation method and application thereof

Abstract

The present invention discloses an amide compound, a preparation method thereof and an application thereof. The structure of the amide compound of the present invention is shown in Formula I-2. The compound of the present invention has a good inhibitory effect on soluble cyclooxygenase.

Description

Technical Field

[0001] The invention relates to an amide compound, a preparation method and application thereof. Background Art

[0002] Epoxide hydrolases (EH, EC 3.3.2.3) catalyze the hydrolysis of epoxides or arene oxides to their corresponding diols (see, Oesch, F. et al., Xenobiotica, 1973, 3, 305-340). Some EHs play a vital role in the metabolism of various compounds, including hormones, chemotherapeutic drugs, carcinogens, environmental pollutants, mycotoxins and other harmful xenobiotics.

[0003] Microsomal epoxide hydrolase (mEH) and soluble epoxide hydrolase (sEH) are two well-studied EHs. In recent years, 1,3-disubstituted ureas, carbamates and amides have been reported as new effective and stable inhibitors of sEH. Chinese patent CN101084216A reported ureas substituted with trifluoromethylphenyl and trifluoromethoxyphenyl. However, more compounds with similar or improved activity need to be developed. Summary of the invention

[0004] The technical problem to be solved by the present invention is the defect that the existing drugs for inhibiting soluble cyclooxygenase have a relatively simple structure. To this end, the present invention provides an amide compound, a preparation method thereof and an application thereof. The compound of the present invention has a good inhibitory effect on soluble cyclooxygenase.

[0005] The present invention also provides a compound as shown in formula I-2 or a pharmaceutically acceptable salt thereof,

[0006]

[0007] wherein ring A is a 3- to 6-membered cycloalkane; n is 0, 1, 2 or 3;

[0008] R3 is independently "C1-C6 alkyl substituted by one or more halogens", "C1-C6 alkoxy substituted by one or more halogens", halogen or sulfur pentafluoride;

[0009] R 2 is hydrogen, C1-C6 alkyl, Phenyl or "phenyl substituted by one or more carboxyl groups"; R 2-1 For hydrogen, C3-C6 cycloalkyl or C1-C6 alkyl; R 2-1-1 It is a C1-C6 alkyl group.

[0010] In a certain embodiment, in the compound of formula I-2 or a pharmaceutically acceptable salt thereof, the definitions of certain groups may be as described below, and the definitions of other groups may be as described in any of the above embodiments (hereinafter referred to as "in a certain embodiment"): wherein ring A is a 3- or 5-membered cycloalkane, n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by 3 halogens", "C1-C6 alkoxy substituted by 3 halogens", halogen or sulfur pentafluoride; 2 Hydrogen or R 2-1 for C3-C6 cycloalkyl or C1-C6 alkyl; R 2-1 - 1 It is a C1-C6 alkyl group.

[0011] In one embodiment, Ring A is a 3- or 5-membered cycloalkane.

[0012] In one embodiment, n is 1 or 2.

[0013] In one scenario, R 3 are independently "C1-C6 alkyl substituted by 3 halogens", "C1-C6 alkoxy substituted by 3 halogens", halogen or sulfur pentafluoride.

[0014] In one scenario, R 2 Hydrogen or

[0015] In one scenario, R 2-1 for C3-C6 cycloalkyl or C1-C6 alkyl.

[0016] In one scenario, R 2-1 for or isopropyl; R 2-1-1 It is methyl.

[0017] In one scheme, when R 3 When it is "C1-C6 alkyl substituted by 3 halogens", R 2 for R 2-1 It is isopropyl.

[0018] In one scheme, when R 3 When it is sulfur pentafluoride, R 2 for R 2-1 for R 2-1-1 It is methyl.

[0019] In one embodiment, the compound shown in formula I-2 is shown in formula I-2-1 Where n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by 3 halogens", "C1-C6 alkoxy substituted by 3 halogens" or halogen; 2 for R 2-1 for C3-C6 cycloalkyl or C1-C6 alkyl; R 2-1-1 It is a C1-C6 alkyl group.

[0020] In one embodiment, the compound shown in formula I-2 is shown in formula I-2-1 Where n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by 3 halogens", "C1-C6 alkoxy substituted by 3 halogens", halogen or sulfur pentafluoride; 2 for R 2-1 for C3-C6 cycloalkyl or C1-C6 alkyl; R 2-1-1 It is a C1-C6 alkyl group.

[0021] In one embodiment, the compound shown in formula I-2 is shown in I-2-2 Where n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by one or more fluorine groups", "C1-C6 alkoxy substituted by one or more halogen groups" or halogen; 2 Hydrogen or R 2-1 It is a C3-C6 cycloalkyl group or a C1-C6 alkyl group.

[0022] In one embodiment, the compound shown in formula I-2 is shown in I-2-2 Where n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by one or more fluorine groups", "C1-C6 alkoxy substituted by one or more halogen groups", halogen or sulfur pentafluoride; 2 Hydrogen or R 2-1 It is a C3-C6 cycloalkyl group or a C1-C6 alkyl group.

[0023] In one embodiment, the compound shown in formula I-2 is shown in formula I-2-3 Where n is 1 or 2; R 2 for R 2-1 for or C1-C6 alkyl; R 2-1-1 is a C1-C6 alkyl group; preferably, sulfur pentafluoride is located at the meta-position and / or para-position of the amide nitrogen.

[0024] In one embodiment, in the compound as shown in Formula I-2, n is 1 or 2; R 3 is sulfur pentafluoride; R 2 for R 2-1 for or C1-C6 alkyl; R 2-1-1 is a C1-C6 alkyl group; preferably, sulfur pentafluoride is located at the meta-position and / or para-position of the amide nitrogen.

[0025] In one embodiment, the compound shown in formula I-2 is shown in formula I-2-4 n is 1 or 2; R 3 are independently "C1-C6 alkyl substituted by 3 halogens" or sulfur pentafluoride; R 2 for R 2-1 for or C1-C6 alkyl; R 2-1-1 It is a C1-C6 alkyl group.

[0026] In one scenario, R 3 are independently "C1-C6 alkyl substituted by 3 halogens" or sulfur pentafluoride.

[0027] In one scenario, R 3 are independently "C1-C6 alkyl substituted by one or more fluorine groups", "C1-C6 alkoxy substituted by one or more halogen groups", halogen or sulfur pentafluoride.

[0028] In one scenario, R 2 for

[0029] In one scenario, R 2-1 for Or C1-C6 alkyl.

[0030] In one scenario, R 2-1 It is a C3-C6 cycloalkyl group or a C1-C6 alkyl group.

[0031] In one embodiment, the sulfur pentafluoride is located in the meta and / or para position relative to the amide nitrogen.

[0032] In one embodiment, in R3, the C1-C6 alkyl group in the “C1-C6 alkyl group substituted by one or more halogens” may be a C1-C4 alkyl group; the C1-C4 alkyl group is preferably a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, a sec-butyl group, an isobutyl group or a tert-butyl group; the C1-C4 alkyl group is also preferably a methyl group; more preferably, the “C1-C6 alkyl group substituted by one or more halogens is a trifluoro-substituted C1-C4 alkyl group”, for example a trifluoromethyl group.

[0033] In one scenario, R 3 In the “C1-C6 alkoxy group substituted by one or more halogens”, the C1-C6 alkoxy group may be a C1-C4 alkoxy group; the C1-C4 alkoxy group is preferably a methoxy group, an ethoxy group, a n-propoxy group, an isopropoxy group, a n-butoxy group, a sec-butoxy group, an isobutoxy group or a tert-butoxy group; the C1-C4 alkoxy group is also preferably a methyloxy group; more preferably, the “C1-C6 alkoxy group substituted by one or more halogens is a trifluoro-substituted C1-C4 alkoxy group, such as a trifluoromethoxy group.

[0034] In one scenario, R 3 In the above, the halogen may be fluorine, chlorine, bromine or iodine; it may also be fluorine.

[0035] In one scenario, R 2 In the above, the C1-C6 alkyl group may be a C1-C4 alkyl group; or may be a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, a sec-butyl group, an isobutyl group or a tert-butyl group.

[0036] In one scenario, R 2-1 In the above, the C1-C6 alkyl group may be a C1-C4 alkyl group; may also be a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, a sec-butyl group, an isobutyl group or a tert-butyl group; may also be an isobutyl group. Or isopropyl.

[0037] In one scenario, R 2-1 In the above, the C3-C6 cycloalkyl group may be a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, and may also be a cyclopropyl group.

[0038] In one scenario, R 2-1-1 In the above, the C1-C6 alkyl group may be a C1-C4 alkyl group; it may also be a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, a sec-butyl group, an isobutyl group or a tert-butyl group; it may also be a tert-butyl group.

[0039] In one scenario, R 2 Can be Boc,

[0040] In one scenario, R 3 It may be fluorine, trifluoromethyl, trifluoromethoxy or sulfur pentafluoride.

[0041] In one plan, Can It can also be

[0042] In one plan, R 3 Located in the meta and / or para position to the amide nitrogen.

[0043] In one embodiment, the compound as shown in formula I-2 is selected from any of the following compounds:

[0044]

[0045]

[0046]

[0047] The present invention also provides a compound as shown in formula I-3 or a salt thereof,

[0048]

[0049] Among them, ring A, n and R 3 As mentioned before.

[0050] In one embodiment, the salt is a salt formed by a conventional inorganic acid in the art and a compound as shown in formula I-3; for example, the salt is a salt formed by the compound shown in I-3 and hydrochloric acid; for another example, the salt is a salt formed by the compound shown in I-3 and hydrochloric acid in a molar ratio of 1:1.

[0051] In one embodiment, the compound as shown in formula I-3 is selected from any of the following compounds:

[0052]

[0053]

[0054] The present invention also provides a method for preparing the compound as shown in Formula I-2, wherein the preparation method is Scheme 1 or Scheme 2.

[0055] The scheme 1 comprises the following steps: in a solvent, a compound as shown in formula II and an amine compound are subjected to a condensation reaction as shown in the following formula in the presence of a base; Ring A, n, R 2 and R 3 The definition of is as mentioned above;

[0056]

[0057] The scheme 2 comprises the following steps: in a solvent, a compound represented by formula I-3 or a salt thereof and a compound represented by formula I-4 are subjected to an amidation reaction represented by the following formula in the presence of a base; R 2 for R 2-1 , Ring A, n and R 3 The definition of is as mentioned above;

[0058]

[0059] In the condensation reaction, the solvent may be a conventional solvent in the art. The solvent may be selected from one or more of an ether solvent, a chlorinated alkane and an aromatic solvent. The ether solvent is preferably diethyl ether, tetrahydrofuran or methyl tert-butyl ether. The chlorinated alkane is preferably dichloromethane. The aromatic solvent is preferably toluene or benzene.

[0060] In the condensation reaction, the amount of the solvent used is the conventional amount in the art, and the volume molar ratio of the solvent to the compound represented by Formula II can be 1-10 mL / mmol; for example, 7 mL / mmol or 8 mL / mmol.

[0061] In the condensation reaction, the molar ratio of the compound represented by Formula II to the amine compound may be 1:(1-3); for example, 1:1.5 or 1:1.7.

[0062] In the condensation reaction, the base may be a conventional organic base in the art, and the base may be a nitrogen-containing organic base; for example, triethylamine.

[0063] In the condensation reaction, the amount of the base used can be the conventional amount in the art; the molar ratio of the compound represented by formula II to the base can be 1:(1-3); for example, 1:1.5 or 1:1.8.

[0064] In the condensation reaction, the reaction temperature of the condensation reaction is a conventional reaction temperature in the art, preferably 0-40°C, such as 0°C, 15°C or 25°C.

[0065] In the scheme 2, the amidation reaction can be carried out in the presence of an acid anhydride. The acid anhydride is a conventional acid anhydride in the art; preferably, the acid anhydride is selected from one or more of propyl phosphoric anhydride, HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) and HOBt (1-hydroxybenzotriazole); for example, propyl phosphoric anhydride. The propyl phosphoric anhydride is preferably added in an ethyl acetate solution with a mass fraction of 50% of propyl phosphoric anhydride.

[0066] In the amidation reaction, the salt is a conventional salt in the art, preferably a hydrochloride.

[0067] In the scheme 2, the amount of the acid anhydride used is the conventional amount in the art, and the molar ratio of the acid anhydride to the compound shown in formula I-3 can be 4:(1-10), such as 3:1, 4:1 or 5:1.

[0068] In the amidation reaction, the molar ratio of the compound represented by Formula II to the compound represented by Formula I-4 can be 1:(1-3); for example, 1:1.3, 1:1.5 or 1:1.7.

[0069] In the amidation reaction, the solvent may be a conventional solvent in the art. The solvent may be a chloroalkane solvent and / or an amide solvent. The chloroalkane solvent is preferably DCM. The amide solvent is preferably DMF.

[0070] In the amidation reaction, the amount of the solvent used is the conventional amount in the art, and the volume molar ratio of the solvent to the compound represented by Formula II can be 1-20 mL / mmol; for example, 11 mL / mmol or 12 mL / mmol.

[0071] In the amidation reaction, the base may be a conventional organic base in the art, and the base may be a nitrogen-containing organic base; for example, triethylamine.

[0072] In the amidation reaction, the amount of the base used can be a conventional amount in the art; the molar ratio of the compound represented by formula II to the base can be 1:(1-10); for example, 1:3, 1:4 or 1:5.

[0073] In the amidation reaction, the reaction temperature of the condensation reaction is a conventional reaction temperature in the art, preferably 0-40°C, such as 0°C, 15°C or 25°C.

[0074] The present invention also provides a use of a substance Z in the preparation of a drug for treating and / or preventing diseases associated with soluble cyclooxygenase; the substance Z is the aforementioned compound as shown in formula I-2 or a pharmaceutically acceptable salt thereof.

[0075] Preferably, the disease is hypertension, pain, cardiomyopathy, inflammation, adult respiratory distress syndrome, diabetic complications, kidney disease, Raynaud's syndrome or arthritis.

[0076] The pain is preferably neuropathic pain and / or inflammatory pain.

[0077] The renal disease is preferably end-stage renal disease.

[0078] The hypertension is preferably renal hypertension, hepatic hypertension or pulmonary hypertension.

[0079] The inflammation is preferably kidney inflammation, vascular inflammation or lung inflammation.

[0080] The present invention also provides an application of a substance Z in the preparation of a soluble cyclooxygenase inhibitor; the substance Z is the aforementioned compound as shown in formula I-2 or a pharmaceutically acceptable salt thereof.

[0081] The present invention also provides a pharmaceutical composition, which comprises a substance Z and a pharmaceutical excipient, wherein the substance Z is the compound shown in the aforementioned formula I-2 or a pharmaceutically acceptable salt thereof.

[0082] In the pharmaceutical composition, the substance Z may be a therapeutically effective amount of substance z.

[0083] Definition of terms

[0084] Certain chemical groups defined herein are preceded by simplified notations to indicate the total number of carbon atoms present in the group. For example, a C1-C6 alkyl group refers to an alkyl group as defined below having a total of 1, 2, 3, 4, 5, or 6 carbon atoms. The total number of carbon atoms in the simplified notation does not include carbons that may be present in substituents of the group.

[0085] The term "room temperature" generally refers to 25-30°C.

[0086] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0087] The term "plurality" refers to 2, 3, 4 or 5.

[0088] The term "pharmaceutically acceptable salt" refers to a salt formed by a suitable non-toxic organic acid, inorganic acid, organic base or inorganic base with a compound of formula I-1, which retains the biological activity of the compound of formula I-1.

[0089] The term "pharmaceutical excipients" refers to excipients and additives used in the production of drugs and the preparation of prescriptions. It is all substances contained in drug preparations except active ingredients. Please refer to the Pharmacopoeia of the People's Republic of China (2015 Edition) Part IV, or Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).

[0090] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) ameliorating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0091] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0092] The term "therapeutically effective amount" refers to an amount of a compound that is sufficient to effectively treat a disease or condition described herein when administered to a patient."Therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as needed by those skilled in the art.

[0093] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0094] The reagents and raw materials used in the present invention are commercially available.

[0095] The positive and progressive effects of the present invention are that the compounds of the present invention have a good inhibitory effect on soluble cyclooxygenase or have good metabolic properties (eg, half-life, exposure AUC, maximum blood drug concentration Cmax). DETAILED DESCRIPTION

[0096] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0097] Chromatographic conditions for preparative separation (purification):

[0098] Separation conditions:

[0099] Mobile phase: Mobile phase A: water (0.225% formic acid); Mobile phase B: acetonitrile

[0100] Chromatographic column: Phenomenexluna C18150*40mm*15μm

[0101] Gradient elution: 37-67% mobile phase B

[0102] Gradient elution time: 10min

[0103] Detection wavelength: 254nm

[0104] Flow rate (ml / min): 60mL / min.

[0105] Example 1 Synthesis of BX20-6-I06

[0106] The synthetic route is shown below:

[0107]

[0108] Compound XR02-S1 (0.27 g, 1.46 mmol), compound Int-1 (0.30 g, 1.33 mmol) and TEA (triethylamine) (0.27 g, 2.66 mmol) were added to DCM (10.0 mL) in sequence, and the reaction mixture was reacted at 15° C. for 12 h. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure at 40° C., and the residue was purified by column chromatography (PE / EA=2 / 1 to 0 / 1, V / V) to obtain a light yellow solid compound BX20-6-I06 (0.27 g, yield 49%).

[0109] 1 H NMR (400MHz, CDCl3) δ = 7.51-7.43 (m, 4H), 7.22 (s, 1H), 5.18 (s, 1H), 4.28 (q, J = 7.2Hz, 1H), 3.56-3.51 (m, 2H), 3.15-3.12 (m, 2H), 2.73 (s, 2H), 1.89-1.67 (m, 4H), 1.45 (s, 9H); MS (ESI, m / z) 358.1[Mt-Bu+H] + .

[0110] Example 2 Synthesis of BX20-6-I07

[0111] Step 1 Synthesis of XR020009-I01

[0112] The synthetic route is shown below:

[0113]

[0114] Compound XR02-S4 (2.00 g, 8.90 mmol) was dissolved in DCM (30 mL), the reaction solution was cooled to below 0°C, BnNH2 (0.95 g, 8.90 mmol) and acetic acid (0.53 g, 8.90 mmol) were added, the reaction solution was placed in an ice bath, stirred for 30 min, NaBH(OAc)3 (3.76 g, 17.8 mmol) in DCM (30 mL) was added, and stirred at 25°C for 12 h. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40°C, and the residue was purified by column chromatography (PE / EA / MeOH=1 / 1 / 0~0 / 5 / 1, V / V) to obtain a light yellow oily compound XR020009-I01 (2.30 g, yield 82.1%). MS (ESI, m / z) 317.2 [M+H] + .

[0115] Step 2: Synthesis of XR020009-I02

[0116] The synthetic route is shown below:

[0117]

[0118] Compound XR020009-I01 (2.30 g, 7.00 mmol) and Pd(OH)2 / C (2.07 g) were used to replace H2, and the reaction mixture was reacted at 25°C for 12 h under H2 (15 psi). After the reaction, the reaction solution was directly filtered and the filtrate was collected. The filtrate was concentrated under reduced pressure at 40°C to obtain a colorless oil compound XR020009-I02 (1.70 g, crude product), which was directly used for the next step reaction, m / z = 227.17 [M+H] + .

[0119] Step 3: Synthesis of compound BX20-6-I07

[0120] The synthetic route is shown below:

[0121]

[0122] Compound XR020009-I02 (1.00 g, 4.40 mmol), XR02-S1 (0.90 g, 4.84 mmol), and TEA (0.90 g, 8.80 mmol) were added to DCM (30 mL) in sequence, and the reaction mixture was reacted at 25° C. for 4 h. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40° C., and the residue was purified by column chromatography (PE / EA=20 / 1 to 1 / 1, V / V) to obtain a white powder compound BX20-6-I07 (1.50 g, yield 83.0%).

[0123] 1 H NMR (400MHz, CDCl3) δ=7.58 (s, 1H), 7.28-7.26 (m, 3H), 5.61 (d, J=7.6Hz, 1H), 4.03-3.94 (m, 1H), 3.25-3.22 (m, 2H), 3.13-3.10(m, 2H), 2.43-2.41(m, 2H), 2.21-2.14(m, 2H), 1.28(s, 9H), 0.95-0.91(m, 2H); MS (ESI, m / z) 414.19[M+H] + .

[0124] Example 3 Synthesis of BX20-6-008

[0125] Step 1: Synthesis of BX20-6-I06

[0126] According to Example 1, a light yellow solid compound BX20-6-I06 was obtained.

[0127] Step 2: Synthesis of compound XR020008-I02

[0128] The synthetic route is shown below:

[0129]

[0130] Compound BX20-6-I06 (0.20 g, 0.48 mmol) was dissolved in anhydrous methanol (5.0 mL), and 2M HCl / EA solution (2.4 mL, 4.80 mmol) was added. The reaction mixture was reacted at 10°C for 5 h. After the reaction was completed, the mixture was concentrated under reduced pressure at 40°C to obtain a white solid compound XR020008-I02 (0.16 g, hydrochloride, crude product), which was directly used in the next step.

[0131] Step 3: Synthesis of compound BX20-6-008

[0132] The synthetic route is shown below:

[0133]

[0134] Compound XR020008-I02 (0.16 g, 0.46 mmol), compound isobutyric acid (52.0 mg, 0.59 mmol), TEA (0.23 g, 2.30 mmol) and T3P (propyl phosphoric anhydride) (1.17 g, 50% T3P mass fraction ethyl acetate solution, 1.84 mmol) were added to DCM (5.0 mL) in sequence, and the reaction mixture was reacted at 15° C. for 6 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure at 40° C., and the residue was purified by preparative separation (formic acid system) and freeze-dried to obtain a white powder compound BX20-6-008 (23 mg, yield 13%).

[0135] 1 H NMR (400MHz, DMSO-d6) δ = 8.74 (s, 1H), 7.59-7.51 (m, 4H), 6.41 (d, J = 7.6Hz, 1H), 4.08 (q, J = 7.2Hz, 1H), 3.65 (t, J = 7.2Hz, 1H), 3.50-3.45 (m, 1H), 3. 28-3.25 (m, 1H), 3.14 (dd, J=8.8, 4.0Hz, 1H), 2.82-2.76 (m, 1H), 2.70-2. 60(m, 2H), 1.80-1.64(m, 4H), 1.00-0.95(m, 6H); MS (ESI, m / z) 384.2[M+H] + .

[0136] Example 4 Synthesis of BX20-6-009

[0137] Step 1: Synthesis of BX20-6-I07

[0138] According to Example 2, a white powdery compound BX20-6-I07 was obtained.

[0139] Step 2: Synthesis of XR020009-I04

[0140] The synthetic route is shown below:

[0141]

[0142] Compound BX20-6-I07 (1.20 g, 2.90 mmol) was dissolved in MeOH (10 mL), HCl (5 mL, 1 M) was added, and the reaction mixture was reacted at 25°C for 12 h. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40°C to obtain a white powder compound XR020009-I04 (1.00 g, crude product, hydrochloride), which was directly used in the next step. MS (ESI, m / z) 314.14 [M+H] + .

[0143] Step 3: Synthesis of BX20-6-009

[0144] The synthetic route is shown below:

[0145]

[0146] Compound XR020009-I04 (0.60 g, 1.71 mmol), isobutyric acid (0.18 g, 2.06 mmol), TEA (0.87 g, 8.58 mmol), and T3P (5.40 g, 8.58 mmol) were added to DCM (20 mL) in sequence, and the reaction mixture was reacted at 25° C. for 12 h. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40° C., and the residue was purified by preparative separation (formic acid system) and freeze-dried to obtain a white powder compound BX20-6-009 (147 mg, yield 22.5%).

[0147] 1H NMR (400MHz, DMSO-d6) δ = 8.72 (s, 1H), 7.57-7.51 (m, 4H), 6.46 (d, J = 7.2Hz, 1H), 4.01-3.94 (m, 1H), 3.61-3.56 (m, 1H), 3.42-3.37 (m, 2H), 2.66-2.61 (m, 2H), 2.53-2.50 (m, 2H), 2.21-2.16 (m, 2H), 1.24-1.14 (m, 2H), 1.00-0.95 (m, 6H); MS (ESI, m / z) 384.1[M+H] + .

[0148] Example 5 Synthesis of BX20-6-011

[0149] Step 1: Synthesis of S2-011

[0150] The synthetic route is shown below:

[0151]

[0152] At -78°C, a solution of compound S1-011 (0.15 g, 0.66 mmol) in DCM (3.0 mL) was slowly added dropwise to a solution of compound Int-1 (0.15 g, 0.75 mmol) and TEA (0.10 g, 1.00 mmol) in DCM (5.0 mL), and the reaction mixture was heated to 20°C and reacted for 0.5 h. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure at 40°C, and the residue was purified by column chromatography (PE / EA=3 / 1~1 / 1, V / V) to obtain a white solid compound S2-011 (160 mg, yield 49.7%), m / z=430.2[M+H] + .

[0153] Step 2: Synthesis of S3-011

[0154] The synthetic route is shown below:

[0155]

[0156] Compound S2-011 (160 mg, 0.37 mmol) was dissolved in MeOH (5.0 mL), and HCl / EA (2 M, 5 mL) was added, and the reaction was stirred at 20°C for 5 hours. After the reaction, the mixture was directly concentrated under reduced pressure at 40°C to obtain yellow solid compound S3-011 (136 mg, crude product, hydrochloride), m / z = 330.3 [M+H] + .

[0157] Step 3: Synthesis of BX20-6-011

[0158] The synthetic route is shown below:

[0159]

[0160] Compound S3-011 (136 mg, crude product, hydrochloride), (S)-(+)-2-methylbutyric acid (46 mg, 0.45 mmol), T3P (50% ethyl acetate solution, 0.76 g, 1.11 mmol) and TEA (187 mg, 1.85 mmol) were added to DMF (2 mL) in sequence, and the reaction mixture was stirred at room temperature for 16 h. After the reaction, water (20 mL) was added, and ethyl acetate (5 mL×3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C, purified by preparative separation (formic acid system), and freeze-dried to obtain compound BX20-6-011 (25 mg, yield 16.3%) as a white powder.

[0161] 1 H NMR (400MHz, DMSO-d6) δ = 8.60 (s, 1H), 7.46 (d, J = 7.6Hz, 2H), 7.20 (d, J = 7.6Hz, 2H), 6.37 (d, J=8.0Hz, 1H), 4.11-4.08 (m, 1H), 3.69-3.66 (m, 1H), 3.52-3.49 (m, 1H), 3.3 1-3.28(m, 1H), 3.20-3.17(m, 1H), 2.86-2.65(m, 2H), 2.49-2.46(m, 1H), 1.80-1.65 (m, 4H), 1.58-1.53 ​​(m, 1H), 1.31-1.23 (m, 1H), 1.00-0.95 (m, 3H), 0.83-0.78 (m, 3H).

[0162] Example 6 Synthesis of BX20-6-012

[0163] Step 1: Synthesis of S2-012

[0164] The synthetic route is shown below:

[0165]

[0166] At -78°C, a solution of compound S1-011 (500 mg, 2.46 mmol) in DCM (5.0 mL) was slowly added dropwise to a solution of compound Int-2 (528 mg, 2.34 mmol) and TEA (28.0 mg, 2.77 mmol) in DCM, and the reaction mixture was reacted at 0°C for 3 h. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure at 40°C, and the residue was purified by column chromatography (PE / EA=3 / 1 to 1 / 1, V / V) to obtain a white solid compound S2-012 (750 mg, yield 71.0%), m / z=430.2 [M+H] + .

[0167] Step 2: Synthesis of S3-012

[0168] The synthetic route is shown below:

[0169]

[0170] Compound S2-012 (750 mg, 1.74 mmol) was dissolved in MeOH (10.0 mL), and HCl / EA (2 M, 20 mL) was added. The reaction was stirred at room temperature for 3 hours. After the reaction was completed, the mixture was directly concentrated under reduced pressure at 40°C to obtain yellow solid compound S3-012 (700 mg, crude product, hydrochloride), m / z = 330.3 [M+H] + .

[0171] Step 3: Synthesis of BX20-6-012

[0172] The synthetic route is shown below:

[0173]

[0174] Compound S3-012 (700 mg, crude product, hydrochloride), (S)-(+)-2-methylbutyric acid (250 mg, 2.46 mmol), T3P (50% ethyl acetate solution, 3.90 g, 6.20 mmol) and TEA (626 mg, 6.20 mmol) were added to DMF (10 mL) in sequence, and the reaction mixture was stirred at room temperature for 16 h. After the reaction, water (80 mL) was added, and ethyl acetate (35 mL×3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C. After purification by preparative separation (formic acid system), and freeze-dried, a white powder compound BX20-6-012 (110 mg, yield 15.3%) was obtained. m / z=414.3[M+H] + .

[0175] 1H NMR (400MHz, DMSO-d6) δ = 8.46 (s, 1H), 7.44 (d, J = 8.8Hz, 2H), 7.18 (d, J = 8.4Hz, 2H), 6.32 (d, J=7.6Hz, 1H), 4.01-3.90 (m, 1H), 3.60-3.55 (m, 1H), 3.42-3.36 (m, 2H), 3.3 3-3.31 (m, 1H), 2.63-2.58 (m, 1H), 2.53-2.50 (m, 1H), 2.49-2.44 (m, 1H), 2.22-2.13 (m, 2H), 1.56-1.48 (m, 1H), 1.31-1.11 (m, 3H), 0.98-0.93 (m, 3H), 0.83-0.76 (m, 3H).

[0176] Example 7 Synthesis of BX20-6-013

[0177] Step 1: Synthesis of XR020008-I02

[0178] According to Example 3, compound XR020008-I02 was synthesized.

[0179] Step 3: Synthesis of BX20-6-013

[0180] The synthetic route is shown below:

[0181]

[0182] Compound XR020008-I02 (135 mg, crude product, hydrochloride), (S)-(+)-2-methylbutyric acid (46 mg, 0.45 mmol), T3P (50% ethyl acetate solution, 0.76 g, 1.11 mmol) and TEA (187 mg, 1.85 mmol) were added to DMF (2 mL) in sequence, and the reaction mixture was stirred at room temperature for 12 h. After the reaction, water (20 mL) was added, and ethyl acetate (5 mL×3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C, purified by preparative separation (formic acid system), and freeze-dried to obtain white powder compound BX20-6-013 (55 mg, yield 33.7%).

[0183] 1H NMR (400MHz, DMSO-d6) δ = 8.74 (s, 1H), 7.58-7.53 (m, 4H), 6.41 (s, 1H), 4.11 -4.09 (m, 1H), 3.69-3.64 (m, 1H), 3.51-3.49 (m, 1H), 3.33-3.30 (m, 1H), 3.2 1-3.16(mm, 1H), 2.83-2.70(m, 2H), 2.49-2.45(m, 1H), 1.79-1.68(m, 4H), 1 .58-1.48(m, 1H), 1.31-1.24(m, 1H), 1.00-0.95(m, 3H), 0.85-0.80(m, 3H).

[0184] Example 8 Synthesis of BX20-6-014

[0185] Step 1 Synthesis of XR020009-I04

[0186] The second step of Example 4 was followed to obtain compound XR020009-I04.

[0187] Step 2 Synthesis of BX20-6-014

[0188] The synthetic route is shown below:

[0189]

[0190] Compound XR020009-I04 (0.35 g, 1.00 mmol), compound (S)-(+)-2-methylbutyric acid (0.13 g, 1.2 mmol), TEA (0.30 g, 3.00 mmol), and T3P (50% ethyl acetate solution, 1.90 g, 3.00 mmol) were added to DMF (5.0 mL) in sequence, and the reaction mixture was reacted at 50° C. for 12 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure at 40° C. to obtain a residue which was purified by preparative separation (formic acid system) and freeze-dried to obtain a white powder compound BX20-6-014 (59.4 mg, yield 15%).

[0191] 1H NMR (400MHz, DMSO-d6) = 8.75 (s, 1H), 7.54 (t, J = 8.0Hz, 4H), 6.50 (d, J = 7.6Hz, 1H), 3.98-3.96 (m, 1H), 3.60-3.56 (m, 1H), 3.42-3.40 (m, 3 H), 2.64-2.61(m, 1H), 2.57-2.51(m, 2H), 2.21-2.16(m, 2H), 1.65-1.47(m, 1H), 1.21-1.16(m, 3H), 0.99-0.94(m, 3H), 0.83-0.78(m, 3H).

[0192] Example 9 Synthesis of BX20-6-015

[0193] Step 1: Synthesis of XR020008-I02

[0194] According to Example 3, compound XR020008-I02 was synthesized.

[0195] Step 3: Synthesis of BX20-6-015

[0196] The synthetic route is shown below:

[0197]

[0198] Compound XR020008-I02 (130 mg, crude product, hydrochloride), cyclopropanecarboxylic acid (70 mg, 0.80 mmol), T3P (50% ethyl acetate solution, 1.27 g, 2.00 mmol) and TEA (160 mg, 1.60 mmol) were added to DMF (3 mL) in sequence, and the reaction mixture was stirred at room temperature for 16 h. After the reaction, water (40 mL) was added, and ethyl acetate (25 mL × 3) was extracted. The organic phase was dried and concentrated under reduced pressure at 40 ° C. After purification by preparative separation (formic acid system), and freeze-dried, a white powder compound BX20-6-015 (55 mg, yield 36.9%) was obtained. m / z = 382.3 [M + H] + .

[0199] 1H NMR (400MHz, DMSO-d6) δ = 8.73 (s, 1H), 7.58-7.53 (m, 4H), 6.42 (d, J = 7.2Hz, 1H), 4.16-4.08 (m, 1H), 3.81 (t, J = 10.8Hz, 1H), 3. 54-3.42 (m, 2H), 3.17 (dd, J=12.0Hz, 4.8Hz, 1H), 2.87-2.82 (m, 1H), 2.74-2.67 (m, 1H), 1.84-1.64 (m, 5H), 0.73-0.68 (m, 4H).

[0200] Example 10 Synthesis of BX20-6-016

[0201] Step 1: Synthesis of XR020009-I04

[0202] The second step of Example 4 was followed to obtain compound XR020009-I04.

[0203] Step 2: Synthesis of BX20-6-016

[0204] The synthetic route is shown below:

[0205]

[0206] At 0°C, compound XR020009-I04 (667 mg, 1.9 mmol, crude product), cyclopropanecarboxylic acid (197 mg, 2.30 mmol), T3P (50% ethyl acetate solution, 3.60 g, 5.70 mmol) and TEA (580 mg, 5.70 mmol) were added to DMF (8 mL) in sequence, and the reaction mixture was stirred at room temperature for 16 h. After the reaction, water (30 mL) was added, and ethyl acetate (50 mL*3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C. The residue was purified by chromatography column (PE / EA / MeOH=1 / 1 / 0~0 / 20 / 1, 1% ammonia water) to obtain white powder compound BX20-6-016 (320 mg, yield 43.9%). m / z=382.2[M+H] + .

[0207] 1H NMR (400MHz, Methanol-d4) δ=7.54-7.49(m, 4H), 4.15-4.13(m, 1H), 3.88-3.84(m, 1H), 3.68-3.62(m, 1H), 3.58-3.55(m , 1H), 3.46-3.41(m, 1H), 2.82-2.66(m, 2H), 2.43-2.32(m, 2H), 1.84-1.77(m, 1H), 1.40-1.28(m, 2H), 0.89-0.80(m, 4H).

[0208] Example 11 Synthesis of BX20-6-017

[0209] Step 1: Synthesis of S2-017

[0210] The synthetic route is shown below:

[0211]

[0212] Triphosgene (59 mg, 0.20 mmol) was dissolved in DCM (5 mL), and after N2 replacement, the temperature was lowered to -78°C, and the reaction mixture of compound S1-017 (86.2 mg, 0.44 mmol) dissolved in DCM (5 mL) was added dropwise to the above reaction system. After the addition, the temperature was naturally raised to 20°C and stirred for 1 h. After the reaction, the reaction solution was directly used for the next reaction.

[0213] Step 2: Synthesis of S3-017

[0214] The synthetic route is shown below:

[0215]

[0216] The reaction solution in Example 1 was cooled to -78°C, Int-1 (150 mg, 0.66 mmol) and TEA (134 mg, 1.33 mmol) were dissolved in DCM (2 mL), and the reaction mixture was slowly added dropwise to the reaction solution in Example 1 and maintained at -78°C. After the addition was completed, the temperature was naturally raised to room temperature and stirred for reaction for 3 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure at 40°C, and the residue was purified by column chromatography (PE / EA=3 / 1~2 / 1, V / V) to obtain a colorless oily compound S3-017 (0.14 g, 73%). MS (ESI, m / z) 448.18 [M+H] + .

[0217] Step 3: Synthesis of S4-017

[0218] The synthetic route is shown below:

[0219]

[0220] Compound S3-017 (0.14 g, 0.32 mmol) and HCl / EA (2 ml, 1.0 M) were dissolved in MeOH (10 mL), and the reaction mixture was stirred at 25°C for 3 h. After the reaction, the reaction mixture was directly concentrated under reduced pressure at 40°C to obtain a colorless oily compound S4-017 (0.10 g, crude product). MS (ESI, m / z) 348.13 [M+H] + .

[0221] Step 4: Synthesis of BX20-6-017

[0222] The synthetic route is shown below:

[0223]

[0224] Compound S4-017 (0.10 g, 0.26 mmol), (S)-(+)-2-methylbutyric acid (0.10 g, 0.94 mmol), T3P (1.2 g, 1.88 mmol), TEA (0.16 g, 1.57 mmol) were added to DMF (3 mL) in sequence, and the reaction mixture was stirred at 25°C for 12 h. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40°C to obtain a residue which was purified by preparative separation (formic acid system) and freeze-dried to obtain a white powder compound BX20-6-017 (12 mg, 16%). MS (ESI, m / z) 432.18 [M+H] + .

[0225] 1 H NMR (400MHz, DMSO-d6) δ = 8.72 (s, 1H), 7.66-7.62 (m, 1H), 7.36 (t, J = 8.0Hz, 1H), 7.08 (d, J = 7.6 Hz, 1H), 6.43-6.42 (m, 1H), 4.08-4.06 (m, 1H), 3.64 (t, J==11.6Hz, 1H), 3.48 (t, J=8.0Hz, 1H), 3 .18-2.17(m, 1H), 3.15-3.14(m, 1H), 2.79-2.76(m, 1H), 2.71-2.66(m, 1H), 2.46-2.43(m, 1H), 1.77-1.67(m, 4H), 1.54-1.46(m, 1H), 1.27-1.23(m, 1H), 0.98-0.93(m, 3H), 0.83-0.78(m, 3H).

[0226] Example 12 Synthesis of BX20-6-018

[0227] Step 1: Synthesis of S1-018

[0228] The synthetic route is shown below:

[0229]

[0230] Triphosgene (345 mg, 1.20 mmol) was added to anhydrous DCM (5.0 mL), nitrogen was replaced, and the reaction solution was cooled to -78 ° C. S1-017 (517 mg, 2.60 mmol) and TEA (400 mg, 4.00 mmol) were dissolved in anhydrous DCM (5.0 mL) to prepare solution A and added dropwise to the reaction solution, and the system was heated to 0 ° C and stirred for 1 hour. The reaction solution was cooled to -78 ° C again, Int-2 (500 mg, 2.20 mmol) and TEA (400 mg, 4.00 mmol) were dissolved in anhydrous DCM (5.0 mL) to prepare solution B and added dropwise to the reaction solution, slowly returned to room temperature and continued to stir for 3 hours. After the reaction, water (30 mL) was added to quench the reaction, and DCM (50 mL*3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C. The residue was purified by chromatography (PE:EA=4:1-1:1) to obtain a white solid s1-018 (600 mg, yield 50.7%), m / z=448.2 [M+H] + .

[0231] Step 2: Synthesis of S2-018

[0232] The synthetic route is shown below:

[0233]

[0234] At room temperature, S1-018 (600 mg, 1.30 mmol) was dissolved in MeOH (3.0 mL), and HCl / EA (2 M, 6.0 mL) was added, and the reaction solution was stirred at 25°C for 2 hours. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40°C to obtain a white solid compound S2-018 (465 mg, crude product, hydrochloride), m / z = 348.1 [M+H] + .

[0235] Step 3: Synthesis of BX20-6-018

[0236] The synthetic route is shown below:

[0237]

[0238] At 0°C, compound S2-018 (465 mg, 1.20 mmol, crude product), (S)-(+)-2-methylbutyric acid (207 mg, 2.0 mmol), T3P (50% ethyl acetate solution, 2.56 g, 4.00 mmol) and TEA (407 mg, 4.00 mmol) were added to DMF (6.0 mL) in sequence, and the reaction mixture was stirred at 25°C for 16 h. After the reaction, water (30 mL) was added, and ethyl acetate (50 mL*3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C. The residue was purified by chromatography (PE: EA = 1: 1 ~ EA: MeOH = 20: 1, 1% ammonia water), and the crude product was separated by preparative separation to obtain white powder compound BX20-6-018 (62 mg, yield 13.5%). m / z = 432.2 [M+H] + .

[0239] 1 H NMR (400MHz, Methanol-d4) = 7.56 (d, J = 12.0Hz, 1H), 7.25 (t, J = 7.6Hz, 1H), 7 .04(d, =12.0Hz, 1H), 4.15-4.06(m, 1H), 3.75-3.69(m, 1H), 3.60-3.52(m, 2H) , 3.49-3.45(m, 1H), 2.79-2.58(m, 3H), 2.41-2.32(m, 2H), 1.69-1.61(m, 1H) , 1.45-1.37(m, 1H), 1.34-1.26(m, 2H), 1.10-1.06(m, 3H), 0.93-0.86(m, 3H).

[0240] Example 13 Synthesis of BX20-6-019

[0241] Step 1: Synthesis of S3-011

[0242] According to Example 8, S3-011 was synthesized.

[0243] Step 2: Synthesis of BX20-6-019

[0244] The synthetic route is shown below:

[0245]

[0246] At 0°C, compound S3-011 (62 mg, 0.20 mmol, crude product), cyclopropanecarboxylic acid (32 mg, 4.00 mmol), T3P (50% ethyl acetate solution, 700 mg, 1.20 mmol) and TEA (76.0 mg, 0.80 mmol) were added to DMF (1.0 mL) in sequence, and the reaction mixture was stirred at 20°C for 12 h. After the reaction, water (5.0 mL) was added, and ethyl acetate (20.0 mL*3) was extracted. The organic phase was dried and concentrated under reduced pressure at 40°C. The residue was separated by preparative separation to obtain white powder compound BX20-6-019 (10 mg, yield 14.9%). m / z=398.2[M+H] + .

[0247] 1 H NMR (400MHz, DMSO-d6) = 8.48 (s, 1H), 7.44 (d, J = 7.6Hz, 2H), 7.19 (d, J = 7.6Hz, 2H), 6.28 (d, J = 4.0Hz, 1H), 4.12-4.08 (m, 1H), 3.79 ( t, J=8.0Hz, 1H), 3.50-3.40(m, 2H), 3.16-3.12(m, 1H), 2.82-2.78(m, 1H), 2.71-2.66(m, 1H), 1.78-1.65(m, 5H), 0.71-0.61(m, 4H).

[0248] Example 14 Synthesis of BX20-6-020

[0249] Step 1: Synthesis of S3-012

[0250] According to the first and second steps of Example 9, compound S3-012 was synthesized.

[0251] Step 2: Synthesis of BX20-6-020

[0252] The synthetic route is shown below:

[0253]

[0254] Compound S3-012 (0.45 g, crude product, hydrochloride), cyclopropanecarboxylic acid (0.21 g, 2.46 mmol), T3P (50% ethyl acetate solution, 3.90 g, 6.20 mmol) and TEA (626 mg, 6.20 mmol) were added to DMF (10 mL) in sequence, and the reaction mixture was stirred at 40°C for 5 h. After the reaction, water (80 mL) was added, and ethyl acetate (35 mL×3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C, purified by preparative separation (formic acid system), and freeze-dried to obtain compound BX20-6-020 (180 mg, yield 36.8%) as a white powder.

[0255] 1 H NMR (400MHz, DMSO-d6) = 8.50 (s, 1H), 7.47 (d, J = 7.6Hz, 2H), 7.20 (d, J = 7.6Hz, 2H), 6.37 (d, J = 4.0Hz, 1H), 4.03-3.99 (m, 1H), 3.74 (t, J = 8.8H z, 1H), 3.57-3.55 (m, 1H), 3.46-3.44 (m, 1H), 2.67-2.53 (m, 2H), 2.23- 2.18 (m, 2H), 1.78-1.78 (m, 1H), 1.30-1.20 (m, 2H), 0.71-0.61 (m, 4H).

[0256] Example 15 Synthesis of BX20-6-021

[0257] Step 1: Synthesis of S3-021

[0258] The synthetic route is shown below:

[0259]

[0260] At 0°C, MeMgCl (3M, 10.4mL, 31.2mmol) was slowly added dropwise to a THF (70mL) solution of S1-021 (4.37g, 25.8mmol) and Ti(OiPr)4 (9.20mL, 30.9mmol). After stirring for 15 minutes, a THF (25mL) solution of S2-021 (6.98g, 30.9mmol) was added. Finally, CyMgBr (2M, 25.8mL, 51.6mmol) was slowly added dropwise at 0°C. After the addition was complete, the reaction was refluxed for 15 minutes, and H2O (18mL) was added after cooling, and stirred at room temperature for 1h. The filtrate was filtered through celite, dried, and concentrated under reduced pressure at 40°C. The residue was purified by column chromatography (PE / EA=10 / 1-5 / 1, V / V) to obtain a light yellow oily compound S3-021 (2.00 g, yield 20.5%), m / z=379.3 [M+H] + .

[0261] Step 2: Synthesis of S4-021

[0262] The synthetic route is shown below:

[0263]

[0264] At room temperature, S3-021 (2.00 g, 5.20 mmol) was dissolved in MeOH (25 mL), Pd / C (10%, 600 mg) was added, and the reaction system was replaced with hydrogen. The mixture was stirred for 48 hours under a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered and directly concentrated under reduced pressure at 40°C to obtain a crude product of a light yellow oily compound S4-021 (1.20 g), m / z = 199.2 [M+H] + .

[0265] Step 3: Synthesis of intermediate S6-021

[0266] The synthetic route is shown below:

[0267]

[0268] At -78°C, a solution of compound S1-011 (700 mg, 3.45 mmol) in DCM (5.0 mL) was slowly added dropwise to a solution of compound S4-021 (500 mg, crude product) and TEA (300 mg, 2.96 mmol) in DCM, and the reaction mixture was reacted at 0°C for 3 h. After the reaction was completed, 2 drops of water were added to the reaction system to quench, and the reaction solution was directly concentrated under reduced pressure at 40°C. The residue was purified by column chromatography (PE / EA=3 / 1-1 / 1, V / V) to obtain a white solid compound S6-021 (525 mg, yield 52.0%), m / z=402.3 [M+H]+ .

[0269] Step 4: Synthesis of S7-021

[0270] The synthetic route is shown below:

[0271]

[0272] Compound S6-021 (500 mg, 1.24 mmol) was dissolved in MeOH (10.0 mL), and then HCl / 1,4-dioxane (4M, 2.00 mL) was added and stirred at room temperature for 3 hours. After the reaction, the mixture was directly concentrated under reduced pressure at 40°C to obtain a white solid compound S7-021 (400 mg, crude product of hydrochloride), m / z=302.3 [M+H] + .

[0273] Step 5: Synthesis of BX20-6-021

[0274] The synthetic route is shown below:

[0275]

[0276] At 0°C, compound S7-021 (400 mg, crude product of hydrochloride), S8-021 (250 mg, 2.50 mmol), T3P (T3P mass fraction 50% ethyl acetate solution, 2.40 g, 4.00 mmol) and TEA (505 mg, 5.00 mmol) were added to DMF (10 mL) in sequence, and the reaction mixture was stirred at room temperature for 16 h. After the reaction, water (80 mL) was added, and ethyl acetate (35 mL×3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C. After purification by preparative separation (hydrochloric acid system), and freeze-dried, a white powder compound BX20-6-021 (133 mg, yield 29.2%) was obtained. m / z=386.2[M+H] + .

[0277] 1H NMR (400MHz, DMSO-d6) δ=8.59 (d, J=3.6Hz, 1H), 7.50 (dd, J=9.2Hz, 2.0Hz, 2H), 7 .22(d, J=8.8Hz, 2H), 6.51(s, 1H), 3.75-3.58(m, 3H), 3.33-3.28(m, 1H), 2.45-2 .38(m,1H),2.23-2.20(m,1H),1.78-1.75(m,1H),1.70-1.66(m,1H),1.56-1.45 (m, 1H), 1.31-1.21 (m, 1H), 0.95 (dd, J=6.8Hz, 2.4Hz, 3H), 0.80 (q, J=7.2Hz, 3H).

[0278] Example 16 Synthesis of BX20-6-022

[0279] Step 1: Synthesis of S3-022

[0280] The synthetic route is shown below:

[0281]

[0282] S1-011226mg, 1.10mmol) was dissolved in anhydrous DCM (1.5mL), nitrogen was replaced, and the solution was cooled to -78°C. S2-022 (200mg, 1.00mmol) and TEA (204mg, 2.00mmol) were dissolved in anhydrous DCM (1.5mL) to prepare solution A, and solution A was added dropwise to the reaction solution. After the addition was completed, the reaction solution was slowly restored to room temperature and stirred for 4 hours. After the reaction was completed, water (30.0mL) was added to quench, and DCM (50.0mL*2) was extracted. The organic phase was dried and concentrated under reduced pressure at 40°C. The residue was purified by chromatography column (PE / EA=5 / 1~2 / 1, V / V) to obtain white solid compound S3-022 (250mg, yield 61.7%), m / z=402.2[M+H] + .

[0283] Step 2: Synthesis of S4-022

[0284] The synthetic route is shown below:

[0285]

[0286] At room temperature, S3-022 (250 mg, 0.60 mmol) was dissolved in MeOH (2.0 mL), and HCl / EA (2 M, 3.0 mL) was added and stirred for 2 hours. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40 °C to obtain a white solid compound S4-022 (188 mg, crude product, hydrochloride), m / z = 302.1 [M+H] + .

[0287] Step 3: Synthesis of BX20-6-022

[0288] The synthetic route is shown below:

[0289]

[0290] At 0°C, compound S4-022 (188 mg, 0.60 mmol, crude product), (S)-(+)-2-methylbutyric acid (128 mg, 1.20 mmol), T3P (T3P mass fraction 50% ethyl acetate solution, 2.40 g, 3.60 mmol) and TEA (252 mg, 2.4. mmol) were added to DMF (4.0 mL) in sequence, and the reaction mixture was stirred at 20°C for 16 h. After the reaction, water (30.0 mL) was added, and ethyl acetate (10.0 mL*3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C. The residue was separated by preparative separation to obtain white powder compound BX20-6-022 (103 mg, yield 42.9%), m / z=386.2[M+H] + .

[0291] 1 H NMR (400MHz, DMSO-d6) = 8.68 (s, 1H), 7.48 (d, J = 8.0Hz, 1H), 7.19 (d, J = 12. 0Hz, 1H), 6.60(s, 1H), 3.73-3.59(m, 3H), 3.28-3.26(m, 1H), 2.43-2.36(m , 1H), 2.19 (d, J=1.6Hz, 1H), 1.75-1.72 (m, 1H), 1.67-1.64 (m, 1H), 1.52-1 .45 (m, 1H), 1.26-1.22 (m, 1H), 0.93 (dd, J=4.0Hz, 3H), 0.80-0.74 (m, 3H).

[0292] Example 17 Synthesis of BX20-6-030

[0293] The synthetic route is shown below:

[0294]

[0295] Triphosgene (368 mg, 1.24 mmol) was added to a three-necked flask, nitrogen was replaced, anhydrous DCM (20 mL) was added, the temperature was lowered to -78 ° C, and a DCM (5.0 mL) solution of XR02-S4 (605 mg, 2.76 mmol) and TEA (1 mL) was slowly added dropwise. After the reaction solution was naturally warmed to -10 ° C, it was continued to stir under an ice bath for 1 hour to obtain a solution of the intermediate XR020007-I01. The temperature was lowered to -78 ° C, and a DCM solution (5 mL) of Int-1 (600 mg, 2.30 mmol) and TEA (1.3 mL) was added dropwise to the DCM solution of the intermediate XR020007-I01. After the addition was completed, the reaction was stirred at room temperature for 1 hour, quenched with water, and concentrated under reduced pressure at 40°C. The residue was purified by column chromatography (PE / EA=3 / 1-1 / 1, V / V) to obtain a crude product (620 mg, yield of 51.5%, purity of 90%). 150 mg of the crude product was purified by preparative separation (formic acid system) and freeze-dried to obtain a white powder compound BX20-6-030 (60 mg, yield of 22.0%). m / z=472.2[M+H] + .

[0296] 1 H NMR (400MHz, DMSO-d6) δ = 8.97 (s, 1H), 7.72 (d, J = 9.2Hz, 2H), 7.55 (d, J = 8.8Hz, 2H), 6.56 (d, J = 7.2Hz, 1H), 4.1 2-4.07 (m, 1H), 3.47-3.42 (m, 2H), 3.04 (dd, J=11.6, 3.6Hz, 2H), 2.72 (s, 2H), 1.76-1.65 (m, 4H), 1.34 (s, 9H).

[0297] Example 18 Synthesis of BX20-6-031

[0298] The synthetic route is shown below:

[0299]

[0300] The compound triphosgene (0.49 g, 1.64 mmol) was dissolved in anhydrous DCM (10 mL) to form mixed solution A, and N2 protection was replaced, and the temperature was lowered to -78°C. XR02-S4 (0.80 g, 3.65 mmol) and TEA (1.5 mL) were dissolved in anhydrous DCM (10 mL) to form mixed solution B. Mixed solution B was taken with a syringe and slowly added to mixed solution A at -78°C. After the addition was completed, the reaction mixture was heated to 0°C and reacted for 1 hour. Int-2 (0.73 g, 3.29 mmol) and TEA (1.5 mL) were dissolved in anhydrous DCM (15 mL), and added dropwise to the above reaction solution AB at -78°C, and stirred for reaction for 2 hours. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40°C, and the residue was purified by silica gel column chromatography (PE:EA=3:1-1:3) to obtain 620 mg of the target product, of which 110 mg was lyophilized to obtain a white powder compound BX20-6-031 (90 mg, 38.7%). m / z=472.1[M+H] + .

[0301] 1 H NMR (400MHz, DMSO-d6) δ = 8.80 (s, 1H), 7.71-7.69 (m, 2H), 7.54-7.52 (m, 2H), 6.47-6.45 (m, 1H), 3.97-3.95 (m , 1H), 3.36-3.43(m, 2H), 3.17-3.14(m, 2H), 2.52(s, 2H), 2.19-2.13(m, 2H), 1.38(s, 9H), 1.23-1.20(m, 2H).

[0302] Example 19 Synthesis of BX20-6-028

[0303] Step 1: Synthesis of BX20-6-030

[0304] According to Example 16, compound BX20-6-030 was synthesized.

[0305] Step 2: Synthesis of XR020028-I01

[0306] The synthetic route is shown below:

[0307]

[0308] Compound BX20-6-030 (470 mg, 0.86 mmol) was dissolved in MeOH (10.0 mL), and HCl / EA (2 M, 2.5 mL) was added, and the reaction was stirred at room temperature for 2 hours. After the reaction, it was directly concentrated under reduced pressure at 40°C to obtain a yellow solid compound XR020028-I01 (450 mg, crude product, hydrochloride), m / z = 372.1 [M+H] + .

[0309] Step 3: Synthesis of BX20-6-028

[0310] The synthetic route is shown below:

[0311]

[0312] Compound (R)-2-methylbutyric acid (60 mg, 0.59 mmol), HATU (220 mg, 0.58 mmol) and TEA (157 mg, 1.55 mmol) were added to DMF (3 mL), stirred at room temperature for 10 minutes, and then XR020028-I01 (200 mg, 0.49 mmol, crude product, hydrochloride) was added. The reaction mixture was stirred at room temperature for 2 h. After the reaction, water (30 mL) was added, and ethyl acetate (25 mL×3) was used for extraction. The organic phase was dried and concentrated under reduced pressure at 40°C. The residue was purified by column chromatography (PE / EA=1 / 1~1 / 3, V / V) to obtain a crude product, which was purified by preparative separation (formic acid system) and freeze-dried to obtain a white powder compound BX20-6-028 (50 mg, yield 33.0%). m / z=456.2[M+H] + .

[0313] 1 H NMR (400MHz, DMSO-d6) δ = 8.88 (s, 1H), 7.74-7.70 (m, 2H), 7.54 (d, J = 8.8Hz, 2H), 6.49 ( d, J=6.8Hz, 1H), 4.13-4.07(m, 1H), 3.69-3.64(m, 1H), 3.52-3.47(m, 1H), 3.32-3.27(m , 1H), 3.19 (dd, J=12.8, 4.8Hz, 1H), 2.82-2.77 (m, 1H), 2.73-2.67 (m, 1H), 1.81-1.66 ( m, 4H), 1.57-1.49 (m, 1H), 1.32-1.23 (m, 2H), 0.97 (q, J=6.8Hz, 3H), 0.85-0.78 (m, 3H).

[0314] Example 20 Synthesis of BX20-6-029

[0315] Step 1: Synthesis of BX20-6-031

[0316] According to Example 17, compound BX20-6-031 was synthesized.

[0317] Step 2: Synthesis of XR020029-I01

[0318] The synthetic route is shown below:

[0319]

[0320] Compound BX20-6-031 (0.51 g, 1.08 mmol) was dissolved in MeOH (10 mL), and HCl / dioxane (2.5 mL, 4 M) was added dropwise. After the addition was completed, the reaction mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure at 40°C to obtain a yellow solid compound XR020029-I01 (0.44 g, crude product). m / z = 408.09 [M+H] + .

[0321] Step 3: Synthesis of BX20-6-029

[0322] The synthetic route is shown below:

[0323]

[0324] Compound (S)-(+)-2-methylbutyric acid (0.13 g, 1.27 mmol), HATU (0.49 g, 1.30 mmol), TEA (0.40 g, 3.90 mmol) were added to DMF (5 mL) in sequence to form mixed solution A. The reaction mixed solution A was stirred at room temperature for 10 min. After the reaction, XR020029-I01 (0.44 g, 1.08 mmol) and TEA (0.40 g, 3.90 mmol) were added to DMF (5 mL) to form mixed solution B. After B was stirred for 5 min, mixed solution B was added dropwise to mixed solution A with a syringe, and the mixture was stirred overnight for 12 h. After the reaction, the reaction solution was directly concentrated under reduced pressure at 40°C to obtain a residue, which was purified by preparative separation (formic acid system) and freeze-dried to obtain a white powder compound BX20-6-029 (270 mg, yield 55%). m / z=456.17[M+H] + .

[0325] 1H NMR (400MHz, DMSO-d6) δ = 8.87-8.86 (m, 1H), 7.73-7.71 (m, 2H), 7.56-7.54 (m , 2H), 6.56-6.54(m, 1H), 4.00-3.98(m, 1H), 3.62-3.57(m, 1H), 3.44-3.39(m, 2H), 2.66-2.62(m, 1H), 2.58-2.53(m, 2H), 2.25-2.16(m, 2H), 1.58-1.50(m, 1 H), 1.38-1.12 (m, 4H), 1.00-0.96 (dd, J=12.0, 4.0Hz, 3H), 0.85-0.78 (m, 3H).

[0326] Example 21 Synthesis of BX20-6-039

[0327] Step 1: Synthesis of S3

[0328] The synthetic route is shown below:

[0329]

[0330] Method: Add bis(trichloromethyl) carbonate (178 mg, 0.6 mmol) to a three-necked flask, replace nitrogen, add anhydrous DCM (10 mL), cool to -78 ° C, and then slowly drop S1 (291 mg, 1.33 mmol) and TEA (403 mg, 3.99 mmol) in DCM (5.0 mL) solution. After the reaction solution is naturally warmed to -10 ° C, continue to stir in an ice bath for 1 hour to obtain a solution of intermediate S2. Cool to -78 ° C, and add Int-1 (300 mg, 1.33 mmol) and TEA (403 mg, 3.99 mmol) in DCM (5 mL) dropwise to the DCM solution of intermediate S2. After the addition is complete, stir the reaction at room temperature for 1 hour. After the reaction, water was added to quench the reaction, and the organic phase was concentrated under reduced pressure after extraction with ethyl acetate. The residue was purified by column chromatography (PE / EA=3 / 1-2 / 1, V / V) to obtain a light yellow solid product S3 (498 mg, yield 79.6%). MS (ESI, m / z) 472.2 [M+H] + .

[0331] Step 2: Synthesis of intermediate S4

[0332] The synthetic route is shown below:

[0333]

[0334] Method: Compound S3 (498 mg, 1.05 mmol) was dissolved in MeOH (10.0 mL), and then HCl / 1,4-dioxane (4 M, 2.5 mL, 10.0 mmol) was added, and the reaction was stirred at room temperature for 3 hours. After the reaction was completed, the mixture was directly concentrated under reduced pressure to obtain a yellow solid S4 (400 mg, crude). MS (ESI, m / z) 372.2 [M+H] + .

[0335] Step 3: Synthesis of BX20-6-039

[0336] The synthetic route is shown below:

[0337]

[0338] Method: At 0°C, compound S4 (400 mg, crude), dimethyl pyrocarbonate (147 mg, 1.1 mmol), and TEA (320 mg, 3.2 mmol) were added to DCM (10 mL), and the reaction mixture was stirred at room temperature for 12 h. After the reaction, the reaction solution was concentrated under reduced pressure and purified by preparative (formic acid system) to obtain a white powder compound BX20-6-039 (295 mg, yield 62.5%). MS (ESI, m / z) 430.1 [M+H] + .

[0339] 1 H NMR(400MHz, METHANOL-d4)δ 7.68-7.64(m, 2H), 7.50(d, J=8.8Hz, 2H), 4.26-4.19(m, 1H), 3.67(s, 3H) , 3.60-3.56(m, 2H), 3.25-3.22(m, 2H), 2.84(brs, 2H), 1.91-1.75(m, 4H).

[0340] Example 22 Synthesis of BX20-6-040:

[0341] Step 1: Synthesis of s3'

[0342] The synthetic route is shown below:

[0343]

[0344] Method: Add bis(trichloromethyl) carbonate (178 mg, 0.6 mmol) to a three-necked flask, replace nitrogen, add anhydrous DCM (10 mL), cool to -78 ° C, and then slowly drop S1 (291 mg, 1.33 mmol) and TEA (403 mg, 3.99 mmol) in DCM (5.0 mL) solution. After the reaction solution is naturally warmed to -10 ° C, continue to stir in an ice bath for 1 hour to obtain a solution of intermediate S2. Cool to -78 ° C, and add Int-1' (300 mg, 1.33 mmol) and TEA (403 mg, 3.99 mmol) in DCM (5 mL) dropwise to the DCM solution of intermediate S2. After the addition is complete, stir the reaction at room temperature for 1 hour. After the reaction was completed, water was added to quench the reaction, and the organic phase was concentrated under reduced pressure after extraction with ethyl acetate. The residue was purified by column chromatography (PE / EA=3 / 1-2 / 1, V / V) to obtain a light yellow solid product S3' (500 mg, yield 79.8%). MS (ESI, m / z) 472.2 [M+H] + .

[0345] Step 2: Synthesis of intermediate S4'

[0346] The synthetic route is shown below:

[0347]

[0348] Method: Compound S3' (500 mg, 1.06 mmol) was dissolved in MeOH (10.0 mL), and then HCl / 1,4-dioxane (4 M, 2.5 mL, 10.0 mmol) was added, and the reaction was stirred at room temperature for 3 hours. After the reaction was completed, the mixture was directly concentrated under reduced pressure to obtain a yellow solid S4' (400 mg, crude product). MS (ESI, m / z) 372.2 [M+H] + .

[0349] Step 3: Synthesis of BX20-6-040

[0350] The synthetic route is shown below:

[0351]

[0352] Method: Compound S4' (400 mg, crude product), dimethyl pyrocarbonate (147 mg, 1.1 mmol), TEA (320 mg, 3.2 mmol) were added to DCM (10 mL) at 0°C, and the reaction mixture was stirred at room temperature for 12 h. After the reaction, the reaction solution was concentrated under reduced pressure and purified by preparative (formic acid system) to obtain a white powder compound BX20-6-040 (323 mg, yield 68.4%). MS (ESI, m / z) 430.2 [M+H]+ .

[0353] 1 H NMR(400MHz, DMSO-d4)δ 8.88 (s, 1H), 7.74-7.70 (m, 2H), 7.56 (d, J=8.8Hz, 2H), 6.51 (d, J=7.6Hz, 1H), 4.03-3.93 (m, 1H), 3.58 (s, 3H), 3.42 ( dd, J=10.8Hz, J=7.6Hz, 2H), 3.23 (dd, J=10.8Hz, J=2.8Hz, 2H), 2.57 (brs, 2H), 2.22-2.15 (m, 2H), 1.27-1.19 (m, 2H).

[0354] Biological test evaluation

[0355] The present invention is further described and explained below in conjunction with test examples.

[0356] Test Example 1 Test of the inhibitory activity of the compounds of the present invention on human soluble cyclooxygenase (IC 50 )

[0357] 1. Purpose of the experiment

[0358] The IC values ​​of the compounds of the present invention against human recombinant soluble cyclooxygenase (sEH) were determined using a sensitive fluorescence-based assay. 50 value.

[0359] 2. Experimental instruments and reagents

[0360] 2.1 Experimental samples

[0361] The examples and reference compounds of the present invention were prepared in-house.

[0362] 2.2 Experimental instruments

[0363] Multifunctional microplate reader (Tecan; Spark20M)

[0364] Microplate constant temperature incubator (Thermo; PST-60HL-4)

[0365] 384-well white plate (Cisbio; 66PL384025)

[0366] 2.3 Experimental reagents

[0367] 3-Phenyl-cyano(6-methoxy-2-naphthyl)methyl ester-2-oxiraneacetic acid (PHOME)(Cayman)

[0368] AUDA (positive compound) (Cayman)

[0369] Tris Buffer (Tris(2-hydroxymethylaminomethane) hydrochloride buffer) (pH 7.0, 25 mM, Sigma)

[0370] Human soluble epoxide hydrolase (hsEH) (Cayman)

[0371] 3. Experimental Procedure

[0372] Determination of IC50 values ​​using a sensitive fluorescence-based assay

[0373] 1) hsEH prepared at 2 μg / mL in Tris Buffer (tris(hydroxymethylaminomethane) hydrochloride buffer) was added to a 384 black plate at 8 μL / well (final concentration 1 μg / mL).

[0374] 2) Then add the prepared compound and positive control to the plate at 4 μL / well. After mixing, centrifuge at 1000 rpm for 1 min. Incubate at 30°C for 5 min. At the same time, set up positive (no compound) and negative control wells (no sEH).

[0375] 3) Then, add the prepared fluorescent substrate PHOME to the plate at 4 μL / well ([S] = 1 μM), mix well, centrifuge at 1000 rpm for 1 min, and incubate at 30°C for 15 min.

[0376] 4) On-machine detection: excitation wavelength: 330nm; emission wavelength: 465nm.

[0377] 4. Data Analysis

[0378] Data were analyzed using GraphPad Prism, and the IC of the compounds was calculated by fitting the percent inhibition and ten-point concentration data to a parametric nonlinear logistic formula. 50 value.

[0379] 5. Experimental Results

[0380] The test results of the inhibitory activity of the compounds of the present invention on human soluble cyclooxygenase are shown in Table 1.

[0381] Table 1

[0382]

[0383]

[0384]

[0385]

[0386] 6. Experimental Conclusion

[0387] The above schemes show that the compounds of the present invention show good inhibitory effects on recombinant human soluble cyclooxygenase.

[0388] Test Example 2 Pharmacokinetics of the compounds of the present invention in SD rats

[0389] 1. Purpose of the experiment

[0390] SD rats were used as test animals to study the pharmacokinetic behavior of the following compound examples in rat plasma after oral administration at a dose of 6 mg / kg.

[0391] 2. Test methods

[0392] 2.1. Test drugs

[0393] The examples and reference compounds of the present invention were prepared in-house.

[0394] 2.2. Experimental animals

[0395] Male SPF SD rats, weighing (200±20) g, from Sibeifu (Beijing) Biotechnology Co., Ltd., animal production license number SCXK (Beijing) 2019-0010.

[0396] 2.3. Preparation of experimental drugs

[0397] Drug preparation: Prepare the drug concentration to 0.3 mg / mL or 0.6 mg / mL, and the prepared solution is 0.5% CMC solution.

[0398] Specific preparation: Weigh 4.5 mg or 9 mg of each drug using a 1 / 10000 precision balance and evenly suspend in 15 mL of 0.5% CMC solution. Be careful to prevent the drug from adhering to the stirring rod or the wall of the test tube during the drug preparation process.

[0399] 2.4. Administration:

[0400] After 3-4 days of adaptive feeding, male SPF SD rats were gavaged and administered with a dose of 6 mg / kg and a dosing volume of 10 mL / kg.

[0401] 2.5. Sample collection

[0402] Before (0h) and after administration, blood was collected from rats at 0.5h, 1h, 1.5h, 2h, 3h, 4h, 6h, 8h, and 24h. Blood was collected by fundus puncture, and the blood volume was about 0.5mL. The blood was placed in a labeled EDTA-K2 anticoagulant tube. Immediately after blood collection, the blood collection tube was gently and completely inverted 3 times to mix with the anticoagulant, and immediately centrifuged at 4500rpm in an ice water bath at 4℃ for 10min. After the centrifugation operation was completed, the plasma was promptly dispensed into EP tubes with corresponding labels and stored in a -80℃ refrigerator.

[0403] 2.6. Sample testing

[0404] This experiment used a simple validated HPLC-MS / MS method to determine the concentrations of Example X and Example Y in plasma after administration.

[0405] 2.6.1. Sample processing: Take 2 μL of blank plasma in a 96-deep-well plate, add 400 μL of precipitant methanol, vortex for 10 min, centrifuge at 4000 rpm for 15 min, take 250 μL of supernatant in a 96-deep-well plate, and perform HPLC-MS / MS analysis with an injection volume of 5 μL.

[0406] 2.6.2. Liquid quality analysis:

[0407] 1) Liquid phase conditions

[0408] Chromatographic column: Waters XSelect CHS C18, 2.1*50mm, 1.8μm; No.: 154;

[0409] Pre-column: Waters in-line filter;

[0410] Mobile phase A: 0.1% formic acid & 5 mM ammonium acetate aqueous solution;

[0411] Mobile phase B: 0.1% formic acid in acetonitrile;

[0412] Flow Rate: 0.6 mL / min;

[0413] Column Temperature: 40°C;

[0414] Sample Tray Temperature: 4°C

[0415] Injection Volume: 5.0 μL;

[0416] Automatic sampling cleaning mode (Rinse Mode): before and after aspiration;

[0417] Automatic sampler needle washing volume (Rinsing Volume): 500.0μL;

[0418] Rinse Dip Time for cleaning the autosampler needle: 5 sec;

[0419] Automatic sampler needle cleaning method (Rinse Method): Rinse Port Only;

[0420] Automatic sampler needle cleaning time (Rinse Time): 2sec

[0421] Rinse Pump and Port Wash Solution: 50% acetonitrile in water.

[0422] Elution method: gradient elution

[0423] 2) Mass spectrometry conditions:

[0424] Ion source: Electrospray Ionization (ESI);

[0425] Ionization mode: positive ion mode (Positive);

[0426] Detection mode: multiple reaction monitoring (MRM);

[0427] Collision cell gas (CAD): 8 psi;

[0428] Curtain Gas Type: 40psi;

[0429] Atomizing gas (GS1): 50psi;

[0430] Auxiliary gas (GS2): 50~60psi;

[0431] Electrospray voltage: 4500 V;

[0432] Turbo Ion Spray Temperature: 600-650°C;

[0433] Mass spectrometry resolution: unit;

[0434] 3. Test results and analysis

[0435] The main pharmacokinetic parameters were calculated using WinNonlin 7.0. The results of the rat pharmacokinetic experiment are shown in Table 2 below.

[0436] Table 2 Pharmacokinetic test results in rats (6 mg / kg dose)

[0437]

[0438]

[0439] 4. Experimental Conclusion

[0440] It can be seen from the rat pharmacokinetic test results in the table that the examples of the present invention exhibit better metabolic properties compared with the reference compounds.

Claims

1. A compound as shown in formula I-2 or a pharmaceutically acceptable salt thereof, characterized in that: wherein ring A is a 5-membered cycloalkane; n is 0, 1, 2 or 3; R 3 are independently "C1-C6 alkyl substituted by 1 or more halogens", "C1-C6 alkoxy substituted by 1 or more halogens", halogen or sulfur pentafluoride; R 2 is hydrogen, C1-C6 alkyl, Phenyl or "phenyl substituted by one or more carboxyl groups"; R 2-1 For hydrogen, C3-C6 cycloalkyl or C1-C6 alkyl; R 2-1-1 It is a C1-C6 alkyl group.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that R 3 In the “C1-C6 alkyl group substituted by one or more halogens”, the C1-C6 alkyl group is a C1-C4 alkyl group; or, R 3 In the "C1-C6 alkoxy group substituted by one or more halogens", the C1-C6 alkoxy group is a C1-C4 alkoxy group; or, R 3 wherein the halogen is fluorine, chlorine, bromine or iodine; or, R 2 wherein the C1-C6 alkyl group is a C1-C4 alkyl group; or, R 2-1 wherein the C1-C6 alkyl group is a C1-C4 alkyl group; or, R 2-1 wherein the C3-C6 cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; or, R 2-1-1 wherein the C1-C6 alkyl group is a C1-C4 alkyl group; Alternatively, the sulfur pentafluoride is located at the meta and / or para position relative to the amide nitrogen.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 3 In the "C1-C6 alkyl group substituted by one or more halogens", the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl; or, R 3 In the "C1-C6 alkoxy group substituted by one or more halogens", the C1-C6 alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy or tert-butoxy; or, R 3 wherein the halogen is fluorine; or, R 2 wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl; or, R 2-1 wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl; or, R 2-1 In the above, the C3-C6 cycloalkyl group is a cyclopropyl group; or, R 2-1-1 In the above, the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 3 In the "C1-C6 alkyl group substituted by one or more halogens", the C1-C6 alkyl group is a methyl group; or, R 3 In the "C1-C6 alkoxy group substituted by one or more halogens", the C1-C6 alkoxy group is a C1-C4 alkoxy group; the C1-C4 alkoxy group is a methyloxy group; or, R 2-1 wherein the C1-C6 alkyl group is isobutyl or isopropyl; or, R 2-1-1 wherein the C1-C6 alkyl group is tert-butyl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 3 In the above, the "C1-C6 alkyl group substituted by one or more halogens" is a trifluoro-substituted C1-C4 alkyl group; or, R 3 In the above, the "C1-C6 alkoxy group substituted by one or more halogens" is a trifluoro-substituted C1-C4 alkoxy group.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 3 In the above, the "C1-C6 alkyl group substituted by one or more halogens" is trifluoromethyl; or, R 3 In the above, the "C1-C6 alkoxy group substituted by one or more halogens" is trifluoromethoxy.

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: n is 1 or 2; or, R 3 are independently "C1-C6 alkyl substituted by 3 halogens", "C1-C6 alkoxy substituted by 3 halogens", halogen or sulfur pentafluoride; or, R 2 Hydrogen or or, R 2-1 for C3-C6 cycloalkyl or C1-C6 alkyl; or, R 2-1-1 It is methyl.

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 3 are independently "C1-C6 alkyl substituted by 3 halogens" or sulfur pentafluoride, or R 3 are independently "C1-C6 alkyl substituted by one or more fluorine groups", "C1-C6 alkoxy substituted by one or more halogen groups", halogen or sulfur pentafluoride; or, R 2 for or, R 2-1 for or C1-C6 alkyl, or R 2-1 It is a C3-C6 cycloalkyl group or a C1-C6 alkyl group.

9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 2-1 for Or isopropyl.

10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: When R 3 When it is "C1-C6 alkyl substituted by 3 halogens", R 2 for R 2-1 isopropyl; and / or, when R 3 When it is sulfur pentafluoride, R 2 for R 2-1 for R 2-1-1 It is methyl.

11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound shown in Formula I-2 is defined in Scheme 1, Scheme 2, Scheme 3, Scheme 4 or Scheme 5: Solution 1: Ring A is a 5-membered cycloalkane, n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by 3 halogens", "C1-C6 alkoxy substituted by 3 halogens", halogen or sulfur pentafluoride; 2 Hydrogen or R 2-1 for C3-C6 cycloalkyl or C1-C6 alkyl; R 2-1-1 is a C1-C6 alkyl group; Solution 2 The compound shown in formula I-2 is shown in I-2-1 Where n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by 3 halogens", "C1-C6 alkoxy substituted by 3 halogens", halogen or sulfur pentafluoride; 2 for R 2-1 for C3-C6 cycloalkyl or C1-C6 alkyl; R 2-1-1 is a C1-C6 alkyl group; Solution 3 The compound as shown in formula I-2 is as shown in I-2-2 Where n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by one or more fluorine groups", "C1-C6 alkoxy substituted by one or more halogen groups", halogen or sulfur pentafluoride; 2 for R 2-1 is C3-C6 cycloalkyl or C1-C6 alkyl; Solution 4: n is 1 or 2; R 3 is sulfur pentafluoride; R 2 for R 2-1 for or C1-C6 alkyl; R 2-1-1 is a C1-C6 alkyl group; Solution 5: n is 1 or 2; R 3 R is independently "C1-C6 alkyl substituted by 3 halogens" or sulfur pentafluoride; 2 for R 2-1 for or C1-C6 alkyl; R 2-1-1 It is a C1-C6 alkyl group.

12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: n is 1 or 2; R 3 is sulfur pentafluoride; R 2 for R 2-1 for or C1-C6 alkyl; R 2-1-1 is a C1-C6 alkyl group; sulfur pentafluoride is located at the meta-position and / or para-position of the amide nitrogen.

13. The compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 For hydrogen, Boc, or, R 3 is fluorine, trifluoromethyl, trifluoromethoxy or sulfur pentafluoride; or, for or, R 3 Located in the meta and / or para position to the amide nitrogen.

14. The compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, characterized in that: for 15. The compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I-2 is selected from any of the following compounds:

16. A compound as shown in formula I-3 or a salt thereof, characterized in that: Among them, ring A, n and R 3 As described in any one of claims 1 to 15.

17. The compound or salt thereof according to claim 16, characterized in that The salt is a salt formed by the compound represented by I-3 and hydrochloric acid.

18. The compound or salt thereof according to claim 16, characterized in that The salt is a salt formed by the compound shown in I-3 and hydrochloric acid in a molar ratio of 1:

1.

19. The compound or salt thereof according to claim 16, characterized in that The compound as shown in formula I-3 is selected from any of the following compounds:

20. A method for preparing a compound as shown in formula I-2, characterized in that: The preparation method is Scheme 1 or Scheme 2, The scheme 1 comprises the following steps: in a solvent, a compound as shown in formula II and an amine compound are subjected to a condensation reaction as shown in the following formula in the presence of a base; Ring A, n, R 2 and R 3 The definition as described in any one of claims 1 to 15; The scheme 2 comprises the following steps: in a solvent, a compound represented by formula I-3 or a salt thereof and a compound represented by formula I-4 are subjected to an amidation reaction represented by the following formula in the presence of a base; R 2 for R 2-1 , Ring A, n and R 3 The definition as described in any one of claims 1 to 15; 21. Use of a substance Z in the preparation of a drug for treating and / or preventing a disease associated with soluble cyclooxygenase; characterized in that: The substance Z is a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

22. The use according to claim 21, characterized in that The disease is hypertension, pain, cardiomyopathy, inflammation, adult respiratory distress syndrome, diabetic complications, kidney disease or Raynaud's syndrome.

23. The use according to claim 22, characterized in that The pain is neuropathic pain and / or inflammatory pain; The kidney disease is end-stage renal disease; The hypertension is renal hypertension, hepatic hypertension or pulmonary hypertension; The inflammation is arthritis, kidney inflammation, vascular inflammation or lung inflammation.

24. Use of a substance Z in the preparation of a soluble cyclooxygenase inhibitor, characterized in that: The substance Z is a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

25. A pharmaceutical composition, characterized in that It comprises a substance Z and a pharmaceutical excipient, wherein the substance Z is a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

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