13-Heterocyclic-β-elemene nitric oxide donor derivatives and their preparation and application

Abstract

The present invention discloses a 13-heterocyclic-β-elemene nitric oxide donor derivative, its preparation, and use. The present invention provides a 13-heterocyclic-β-elemene nitric oxide donor derivative having a structure as shown in general formula (I), or a pharmaceutically acceptable salt or solvate thereof, or an enantiomer or diastereomer thereof. The derivative of the present invention surpasses previous β-elemene nitric oxide donor derivatives in design strategy by introducing a nitrogen-containing ring structure and an NO donor that enhance antitumor activity in vivo. This significantly improves the physicochemical properties of the β-elemene nucleus and can be used to treat leukemia, among other diseases for which effective therapeutic agents are currently lacking. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the fields of medicinal chemistry and new drug research and development, and in particular to 13-heterocyclic-β-elemene nitric oxide donor derivatives and preparation and application thereof. Background Art

[0002] Elemene, a natural sesquiterpene product extracted and isolated from Curcuma zedoaria, is an anticancer botanical drug that does not contain toxic groups such as anthracene, epoxy, or nitro groups. Elemene oral emulsion and injection, approved by the China Food and Drug Administration (NMPA), have been used clinically for many years as a broad-spectrum anticancer agent. Elemene extracts contain multiple isomers, of which β-elemene exerts the most significant antitumor activity. β-elemene is a small molecule compound, consisting of only carbon and hydrogen. This results in poor water solubility and low bioavailability, requiring high doses to be effective, limiting its clinical application. Therefore, structural optimization of elemene is of great research significance.

[0003] Nitric oxide (NO), an endogenous gas molecule, participates in various physiological and pathological processes, including anti-inflammatory and anti-tumor activities. Compared to normal cells, tumor cells are more sensitive to NO. Low concentrations of NO can promote tumor cell growth, but high concentrations can reverse tumor cell drug resistance and inhibit tumorigenesis and metastasis. NO has a short half-life, is small, and is lipophilic, allowing it to rapidly permeate cell membranes. However, it is difficult to quantify and transport. Therefore, NO donors are used clinically as NO substitutes.

[0004] Compared to traditional nitrate NO donors, furazans offer the advantage of releasing NO through non-enzymatic pathways in most tissues and organs, generating biological activity. Most small-molecule NO donors are unstable and catalytically degrade during blood circulation, resulting in insufficient concentrations before reaching target cells or tissues, potentially promoting tumor cell growth and achieving the opposite effect. Therefore, using NO donors directly to treat hematologic cancers (such as leukemia) offers greater targeting and potentially improved efficacy.

[0005] Furthermore, our research has found that introducing nitrogen-containing heterocyclic structures, such as pyrazole and N-ethylpiperazine, into the β-elemene structure can significantly enhance its antitumor activity. Furthermore, we chose to introduce an alcoholamine linker containing an amino group and an alcoholic hydroxyl group at position 14 of β-elemene, which is then linked to a furazan-type NO donor. Therefore, based on the principle of pharmacophore fusion, the present invention fuses β-elemene with both the NO pharmacophore and the nitrogen-containing heterocyclic pharmacophore within the β-elemene molecule, significantly improving its physicochemical and druggability, resulting in a β-elemene derivative with enhanced antitumor efficacy. Summary of the Invention

[0006] The first objective of the present invention is to address the deficiencies in the prior art and provide a 13-heterocyclic-β-elemene nitric oxide donor derivative, wherein a nitrogen-containing cyclic group is introduced at the 13th position of elemene, and then the 14th position of elemene is connected to a furazan nitric oxide donor using an alcohol amine and an acid anhydride as a linker arm, thereby finally preparing a β-elemene nitric oxide donor derivative with a novel linker arm, which has excellent anti-tumor activity.

[0007] A 13-heterocyclic-β-elemene nitric oxide donor derivative, or a pharmaceutically acceptable salt or solvate thereof, or an enantiomer or diastereomer thereof, wherein the 13-heterocyclic-β-elemene nitric oxide donor derivative has the following general formula (I):

[0008]

[0009] in:

[0010] R 1 It is a ring structure containing nitrogen, oxygen or sulfur;

[0011] R 2 It is a straight-chain or cyclic alcohol amine structure containing nitrogen and oxygen atoms;

[0012] R 3 、R 4 Each independently selected from C 1-10 Alkyl, C 3-12 Cycloalkyl, C 6-12 Aryl, 5-10 membered heteroaryl, C 2-10 Alkenyl, C 2-10 Alkynyl or C 2-10 Alkoxy.

[0013] Preferably, in formula (I):

[0014] R 1 C 2-8 Ring structures containing nitrogen, oxygen or sulfur;

[0015] R 2 C 2-5 The structure of a straight-chain alcohol amine containing nitrogen and oxygen atoms or C 5-8 Cyclic alcohol amine structure;

[0016] R 3 、R 4 Each independently selected from C 2-6 Chain alkyl, C 2-6 Alkenyl, C 2-6 of alkynyl.

[0017] Preferably, in formula (I):

[0018] R 1 Selected from

[0019] R 2 Selected from The oxygen atom is attached to the carbonyl group;

[0020] R 3 、R 4 They are independently selected from -CH2CH2-, -CH=CH-, -CH2CH2CH2-, -CH2CH=CH-, -CH=CHCH2-, -CH2C≡C-, -C≡CCH2-, -CH2CH2CH2CH2-, -CH2CH2CH=CH-, -CH2CH=CHCH2-, -CH=CHCH2-, -CH2C≡CCH2-, -CH2CH2CH2CH2CH2-, -CH2C≡CCH2CH2-, -CH2CH2C≡CCH2-, -CH2CH2C≡CCH2-, and are connected to the two end structures from left to right.

[0021] Preferably, in formula (I):

[0022] R 1 for

[0023] R 2 Selected from The oxygen atom is attached to the carbonyl group;

[0024] R 3 、R 4 They are independently selected from -CH2CH2-, -CH=CH-, -CH2CH2CH2-, -CH2CH=CH-, -CH=CHCH2-, -CH2C≡C-, -C≡CCH2-, -CH2CH2CH2CH2-, -CH2CH2CH=CH-, -CH2CH=CHCH2-, -CH=CHCH2-, -CH2C≡CCH2-, -CH2CH2CH2CH2CH2-, -CH2C≡CCH2CH2-, -CH2CH2C≡CCH2-, -CH2CH2C≡CCH2-, and are connected to the structures at both ends in order from left to right.

[0025] Preferably, the 13-heterocyclic-β-elemene nitric oxide donor derivative is selected from compounds 1 to 36 shown in the following structures:

[0026]

[0027]

[0028]

[0029] The second object of the present invention is to provide a method for preparing the above-mentioned 13-heterocyclic-β-elemene nitric oxide donor derivative, using the following technical scheme:

[0030] (1) β-elemene A-1 undergoes allylic chlorination at positions 13 and 14 to obtain intermediate A-2;

[0031] (2) R containing nitrogen heteroatom functional groups 1 Structural fragment A-3 is selectively substituted into position 13 of intermediate A-2 to give intermediate A-4;

[0032] (3) R containing nitrogen heteroatom functional groups 2 Structural fragment A-5 is selectively substituted into position 14 of intermediate A-4 to give intermediate A-6;

[0033] (4) Phenylthioacetic acid A-7 is oxidized with 30% H2O2 and then treated with fuming nitric acid to obtain intermediate A-8;

[0034] (5) R containing hydroxyl functional group 3 Structural fragment A-9 is selectively nucleophilically substituted onto intermediate A-10 to obtain intermediate A-10;

[0035] (6) R containing a carbonyl functional group 4 Structural fragment A-11 is esterified with intermediate A-10 to obtain intermediate A-11;

[0036] (7) Intermediate A-6 and intermediate A-11 undergo intermolecular amide condensation to obtain a derivative represented by general formula (I);

[0037] Its synthetic route is as follows:

[0038]

[0039] The third object of the present invention is to provide the use of the 13-heterocyclic-β-elemene nitric oxide donor derivative, or its pharmaceutically acceptable salt, or solvate, or its enantiomer or diastereomer in the preparation of anti-tumor drugs.

[0040] Preferably, the tumor includes leukemia, melanoma, etc.

[0041] The fourth object of the present invention is to provide an anti-tumor drug containing a safe and effective amount of the 13-heterocyclic-β-elemene nitric oxide donor derivative, or a pharmaceutically acceptable salt or solvate thereof, or an enantiomer or diastereomer thereof.

[0042] Preferably, the anti-tumor drug may further include a pharmacologically acceptable salt and a pharmacologically acceptable excipient or carrier.

[0043] As used herein, a "safe and effective amount" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, a pharmaceutical composition contains 1-2000 mg of the compound of this invention per dose, more preferably 5-1000 mg per dose. Preferably, a "dose" is one capsule or tablet.

[0044] "Pharmaceutically acceptable carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0045] In the present invention, the synthesis route of 13-heterocyclic-β-elemene is shown in Reaction Formula 1.

[0046]

[0047] Compared with the prior art, the present invention adopts 13-heterocyclic-β-elemene as an anti-tumor active skeleton, and has the following important advantages: 1) in the chlorination reaction of β-elemene, 13,14-dichloro-elemene is generated as the main product, the yield of monosubstituted elemene is very low, the atom utilization rate is greatly improved, the subsequent separation and purification are easier than the monosubstituted elemene, and it is more suitable for industrial large-scale production, which provides convenience for the subsequent research and development of such drugs; 2) the anti-tumor activity of 13,14-dichloro-β-elemene is equivalent to that of 13-chloro-β-elemene, 14-chloro-β-elemene and β-elemene. On the one hand, the polarity of the dichloro compound is greater than that of the monosubstituted compound. On the other hand, the compounds chlorinated at the 13 and 14 positions can introduce different groups to exert different biological activities and show better drug-like properties.

[0048] At the same time, the present invention introduces a nitrogen-containing heterocycle and an NO donor as a pharmacophore that can enhance the anti-tumor activity in vivo, thereby improving the physical and chemical properties of the elemene parent body; therefore, the β-elemene nitric oxide donor derivative disclosed in the present invention is easy to prepare and separate, has better safety for long-term medication, can quickly release NO in the blood, exhibits better targeting and therapeutic activity against blood diseases such as leukemia, and is highly innovative and novel. DETAILED DESCRIPTION

[0049] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the invention. The operating methods in the following examples where no specific conditions are specified are generally performed under conventional conditions or as recommended by the manufacturer.

[0050] (I) Preparation of intermediate 2

[0051]

[0052] To a mixed solution of β-elemene (29.4 mmol) in dichloromethane (40 mL) and acetic acid (35 mL) was added TBAF (1.0 min THF, 0.5 mL) at 0°C. NaClO (40 mL) was then added using a microsyringe pump, and the mixture was allowed to react at 0°C for 5 h. Upon completion of the reaction, the mixture was quenched with saturated sodium bicarbonate and extracted with ethyl acetate. The organic phases were combined, washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was separated by silica gel column chromatography (pure petroleum ether) to obtain a colorless liquid compound, intermediate 2, with a yield of 25%.

[0053] 1 HNMR(500MHz, CDCl3)δ5.79(dd,J=17.2,11.0Hz,1H),5.28(s,1H),5.18(s,1H),5.04(s,1H),4.97–4 .89(m,3H),4.11(s,3H),3.97(d,J=11.7Hz,1H),2.35–2.22(m,2H),1.77–1.42(m,9H),0.99(s,3H). 13 CNMR (125MHz, CDCl3) δ149.51,148.95,147.44,116.29,113.33,111.42,50.88,47.63,47.56,41.01,39.72,39.61,33.76,26.97,15.73.

[0054] (II) Preparation of Intermediate 3-4

[0055]

[0056] Reaction Scheme 2: Synthesis of Intermediates 3-4 and 5-10. Reagents and conditions: (a) Cs2CO3, DMF, rt,

[0057] 12h; (b) Cs2CO3, DMF, 60°C.

[0058] As shown in Reaction Scheme 2, Cs2CO3 (2.0 mmol) was added to DMF (6 mL) containing Intermediate 2 (1.0 mmol) and stirred at room temperature for 20 min. Pyrazole (2.0 mmol) was added to the mixed solution and stirred at room temperature for 12 h. Upon completion of the reaction, the mixed solution was diluted with water and extracted with ethyl acetate. The combined organic solutions were washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, and the residue was separated by silica gel column chromatography (dichloromethane:methanol volume ratio 100:1) to obtain Intermediate 3 (67%) as a colorless, transparent liquid.

[0059] 1 HNMR (500MHz, CDCl3) δ7.52(d,J=1.6Hz,1H),7.40(d,J=2.2Hz,1H),6.28(t,J=2 .0Hz,1H),5.74(dd,J=10.9,17.3Hz,1H),5.25(s,1H),5.02(s,1H),4.94–4.86( m,3H),4.78(s,3H),4.06(d,J=12.5Hz,1H),3.94(d,J=11.7Hz,1H),2.22(dd,J= 3.6,12.4Hz,1H),1.95(td,J=3.5,11.7Hz,1H),1.65–1.44(m,6H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ149.5,149.0,147.4,139.3,129.5,116.4,111.8,111.5,10 5.9,56.3,51.0,47.6,41.9,39.8,39.6,33.7,26.8,15.8.MS(ESI)m / z:305.2[M+H] + .

[0060] Following the same method, by replacing the reactants, intermediate 4 was obtained.

[0061] 1-(2-((1R,3R,4S)-3-(3-chloroprop-1-en-2-yl)-4-methyl-4-vinylcyclohexyl)allyl)-4-ethylpiperazine (4)

[0062] Colorless transparent liquid, yield 59%, 1HNMR (500MHz, CDCl3) δ5.71–5.64(m,1H),5.16(s,1H),4.82(dd,J=3.6,15.5Hz,5H),3.99(d,J=12.2Hz,1H),3.86(d,J=11.7Hz,1H),2 .86–2.79(m,2H),2.36(q,J=7.2Hz,11H),2.19–2.15(m,1H),2.04–1.98(m,1H),1.54–1.33(m,6H),1.00(d,J=7.2Hz,3H),0.87(s,3H). 13 CNMR (125MHz, CDCl3) δ150.8,149.4,147.8,116.2,111.2,110.9,63.5,53.3,53.1, 52.5,51.1,47.9,42.3,39.9,39.9,34.0,27.1,15.9,12.1.MS(ESI)m / z:351.2[M+H] + .

[0063] (III) Preparation of Intermediate 5-10

[0064] To a solution of 3 (1.0 mmol) in DMF (5 mL) were added N-methyl-2-hydroxyethylamine (1.5 mmol) and DIPEA (2.0 mmol), followed by reaction at 60°C for 12 h. After completion of the reaction, the mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was chromatographed on silica gel (dichloromethane:methanol, volume ratio, 60:1) to afford intermediate 5 (51%).

[0065] 1 HNMR(500MHz, CDCl3)δ7.50(d,J=1.5Hz,1H),7.39(d,J=2.1Hz,1H),6.26(t,J=2.0Hz ,1H),5.74(dd,J=17.8,10.5Hz,1H),5.01(d,J=17.9Hz,2H),4.90–4.85(m,2H),4.79 –4.70(m,4H),3.62–3.54(m,2H),3.13(d,J=13.4Hz,1H),2.67–2.54(m,2H),2.35(dt ,J=12.4,4.8Hz,1H),2.15(s,4H),1.92–1.82(m,1H),1.60–1.39(m,6H),0.96(s,3H). 13CNMR (125MHz, CDCl3) δ149.7,149.6,148.5,139.2,129.4,114.6,111.5,110.5,105.8,66.1 ,58.6,58.5,56.3,47.6,41.9,41.8,39.7,39.6,33.8,26.8,15.9.MS(ESI)m / z:344.2[M+H] + .

[0066] By the same method, replacing the reactants, intermediate 6-10 was prepared.

[0067] (1-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl)-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methanol (6)

[0068] Light yellow liquid, yield 46%, 1 HNMR (500MHz, CDCl3) δ7.51(d,J=1.4Hz,1H),7.39(d,J=1.8Hz,1H),6.27(t,J=2.0Hz,1H),5 .79(dd,J=10.8,17.5Hz,1H),5.03(d,J=18.8Hz,2H),4.89–4.83(m,2H),4.80–4.72(m,4H),3 .55–3.45(m,2H),3.09–2.95(m,1H),2.92–2.76(m,2H),2.62(s,1H),2.24–2.12(m,1H),2.0 8–1.86(m,2H),1.75–1.63(m,3H),1.50(dddd,J=12.2,23.2,34.2,44.0Hz,9H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ149.8,149.7,148.3,139.1,129.4,113.5,111.5,110.2,105.7,67.6,66.6,5 6.2,54.4,52.7,47.9,41.9,39.7,38.7,33.6,29.7,28.9,28.8,26.9,15.9.MS(ESI)m / z:384.2[M+H] + .

[0069] 2-(4-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl)-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethan-1-ol (7)

[0070] Copper green liquid, yield 80%, 1 HNMR (500MHz, CDCl3) δ7.49 (d, J = 1.5 Hz, 1H), 7.37 (d, J = 2.1 Hz, 1H), 6.25 (t, J = 2. 0Hz,1H),5.75(dd,J=10.8,17.5Hz,1H),5.00(d,J=9.4Hz,2H),4.89–4.82(m,2H) ,4.75(s,4H),3.60(t,J=5.4Hz,2H),3.00(d,J=13.7Hz,1H),2.63–2.22(m,12H), 2.14(dd,J=3.4,12.6Hz,1H),1.93–1.84(m,1H),1.61–1.38(m,6H),0.95(s,3H). 13 CNMR (125MHz, CDCl3) δ149.9,149.8,148.2,139.3,129.4,113.9,111.6,110.3,105.8,6 6.2,59.5,57.8,56.4,53.1,47.9,41.9,39.8,33.7,27.1,16.0.MS(ESI)m / z:399.2[M+H] + .

[0071] 2-((2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl)-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethan-1-ol (8)

[0072] Light yellow liquid, yield 66%, 1 HNMR (500MHz, CDCl3) δ5.71–5.64(m,1H),5.16(s,1H),4.82(dd,J=3.6,15.5Hz,5H),3.99(d,J=12.2Hz,1H),3.86(d,J=11.7Hz,1H),2 .86–2.79(m,2H),2.36(q,J=7.2Hz,11H),2.19–2.15(m,1H),2.04–1.98(m,1H),1.54–1.33(m,6H),1.00(d,J=7.2Hz,3H),0.87(s,3H). 13CNMR (125MHz, CDCl3) δ150.9,150.1,149.1,114.3,110.9,110.4,66.3,63.7,59.0,58.6,53. 3,53.0,52.4,48.0,42.4,41.9,40.0,39.9,34.3,27.2,16.0,12.1.MS(ESI)m / z:390.4[M+H] + .

[0073] (1-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl)-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methanol (9)

[0074] Colorless transparent liquid, yield 64%, 1 HNMR(500MHz, CDCl3)δ5.81(dd,J=10.8,17.5Hz,1H),5.03(s,1H),4.95–4.83(m,4H),4.77(s,1H),3.49(d,J=6.4Hz,2H),3.04–

[0075] 2.82(m,5H),2.66–2.32(m,10H),2.22(dd,J=3.2,12.7Hz,1H),2.12–1.91(m,3H),1.75–1.39(m,11H),1.10(t,J=7.2Hz,3H),0.99(s,3H). 13 CNMR (125MHz, CDCl3) δ151.1,150.3,149.0,113.2,110.7,110.0,67.9,66.8,63.6,54.5,53.2,53.0, 52.8,52.4,48.3,42.4,40.1,40.0,38.9,34.1,29.1,29.0,27.3,16.1,12.0.MS(ESI)m / z:430.4[M+H] + .

[0076] 2-(4-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl)-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethan-1-ol (10)

[0077] Light yellow liquid, yield 76%, 1HNMR(500MHz, CDCl3)δ5.81(dd,J=10.8,17.5Hz,1H),5.03(s,1H),4.95–4.83(m,4H),4.77(s,1H),3.49(d,J=6.4Hz,2H),3.04–2.82 (m,5H),2.66–2.32(m,10H),2.22(dd,J=3.2,12.7Hz,1H),2.12–1.91(m,3H),1.75–1.39(m,11H),1.10(t,J=7.2Hz,3H),0.99(s,3H). 13 CNMR (125MHz, CDCl3) δ151.0,150.2,148.5,113.6,110.7,110.1,66.3,63.6,59.5,57.8,53. 2,53.1,53.0,52.4,48.2,42.4,40.0,39.9,34.1,27.2,16.0,12.0.MS(ESI)m / z:445.4[M+H] + .

[0078] (IV) Preparation of Intermediates 13-15

[0079]

[0080] Reaction Scheme 3. Synthesis of Intermediates 13-15 and 13a-b, 14a-b, 15a-b. Reagents and Conditions: (a) 15%

[0081] NaOH, THF, rt, 2h; (b) DMAP, DCM, rt, 12h.

[0082] To a solution of 3,4-bis(phenylsulfonyl)-1,2,5-oxadiazole-2-oxide (2.0 mmol) in tetrahydrofuran was slowly added dropwise with a 15% aqueous NaOH solution (0.4 mmol) at 0°C. After the addition was complete, stirring was continued for 10 minutes. Ethylene glycol (20.0 mmol) was then added and the reaction was continued at room temperature for 4 hours. The solvent was removed by distillation under reduced pressure, the mixture was diluted with water, and extracted with dichloromethane. The organic phases were combined, washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was chromatographed on silica gel (dichloromethane:methanol, volume ratio, 100:1) to afford Intermediate 13 (77%).

[0083] mp128-130℃. 1 HNMR(400MHz, CDCl3)δ8.07(d,J=8.2Hz,2H),7.77(t,J=

[0084] 7.5Hz,1H),7.63(t,J=7.8Hz,2H),4.59–4.49(m,2H),4.08–4.02(m,2H). 13 CNMR(100MHz,CDCl3)δ159.1,137.8,135.9,129.9,128.7,110.7,73.0,60.5.HRMS(ESI)calcdforC 10 H 10 N2NaO6S309.0152[M+Na] + ,found309.0142.

[0085] According to Reaction Scheme 3 and the same method as above, intermediates 14 and 15 were prepared by replacing the reactants.

[0086] 4-(3-Hydroxypropoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide (14)

[0087] White solid, yield 73%, mp109-110℃. 1 HNMR (500MHz, CDCl3) δ8.05 (dd, J=8.5, 1.1Hz, 2H), 7.78–7.73 (m, 1H), 7.64–7. 60(m,2H),4.59(t,J=6.0Hz,2H),3.87(t,J=5.8Hz,2H),2.13(p,J=5.9Hz,2H). 13 CNMR(125MHz, CDCl3)δ159.0,135.8,129.8,110.6,69.4,59.4,31.4.MS(ESI)m / z:323.0[M+Na] + .

[0088] 4-((4-Hydroxybutyl 2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide (15)

[0089] White solid, yield 59%, mp116-118℃. 1 HNMR (400MHz, CDCl3) δ8.07(d,J=7.7Hz,2H),7.76(t,J=7.4Hz,1H),7.63(t,J=7.7Hz,2H),5.10(s,2H),4.34(s,2H). 13 CNMR(100MHz, CDCl3)δ158.2,138.0,135.9,129.8,128.8,110.8,88.3,77.7,59.0,51.1.HRMS(ESI)calcdforC12 H 10 N2NaO6S333.0512[M+Na] + ,found333.0512.

[0090] (V) Preparation of Intermediates 13a, 13b, 14a, 14b, 15a, and 15b

[0091] To a solution of Intermediate 13 (2.0 mmol) in dichloromethane (10 mL) were added succinic anhydride (2.2 mmol) and DMAP (1.0 mmol), followed by stirring at 30°C for 3 h. The reaction was monitored by thin-layer chromatography. The reaction mixture was diluted with dichloromethane. The organic phase was washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane:methanol volume ratio 100:1) to afford a white solid, Intermediate 13a (58%).

[0092] mp118-120℃. 1 HNMR(400MHz, CDCl3)δ8.13–8.06(m,2H),7.82–7.76(m,1H),7.65(t,J=7.9Hz,2H ), 4.65 (dd, J = 5.4, 3.4Hz, 2H), 4.56 (dd, J = 5.4, 3.4Hz, 2H), 2.73 (t, J = 2.7Hz, 4H). 13 CNMR(100MHz, CDCl3)δ176.9,171.9,158.8,138.2,135.8,129.8,128.8,110.6,69.0,61.6,29.0,28.9.HRMS(ESI)calcdforC 14 H 14 N2NaO9S409.0312[M+Na] + ,found409.0295.

[0093] By following the same method as above, replacing the reactants, intermediates 13b, 14a, 14b, 15a, and 15b were obtained.

[0094] Intermediates 13b, 14a, 14b, 15a, 15b 1 HNMR

[0095] 4-(2-((4-carboxybutyryl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole-2-oxide (13b)

[0096] White solid, yield 63%, mp 91-93 ℃. 1HNMR(500MHz, CDCl3)δ8.06(dd,J=8.5,1.2Hz,2H),7.78–7.73(m,1H),7.62(td,J=7.6,1.7Hz,2H), 4.65–4.61(m,2H),4.51(dd,J=5.4,3.7Hz,2H),2.47(dt,J=8.3,7.3Hz,4H),1.99(p,J=7.2Hz,2H). 13 CNMR(100MHz, CDCl3)δ178.6,172.7,158.8,138.0,135.8,129.8,128.8,110.6,69.0,61.3,33.0,32.9,19.7.HRMS(ESI)calcdforC 15 H 16 N2NaO9S423.0469[M+Na] + ,found423.0465.

[0097] 4-(3-((3-carboxypropionyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide (14a)

[0098] White solid, yield 77%, mp 104-105℃, 1 HNMR(500MHz, CDCl3)δ8.05(dd,J=8.5,1.2Hz,2H),7.78–7.73(m,1H),7.65–7.59(m,2H) ,4.50(t,J=6.1Hz,2H),4.31(t,J=6.1Hz,2H),2.70–2.60(m,4H),2.22(p,J=6.1Hz,2H). 13 CNMR(125MHz, CDCl3)δ178.0,172.2,159.0,138.1,135.8,129.8,128.6,110.6,67.9,60.7,28.9,28.0.MS(ESI)m / z:423.0[M+Na] + .

[0099] 4-(3-((4-carboxybutyryl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide (14b)

[0100] White solid, yield 77%, mp96-98℃, 1HNMR(500MHz, CDCl3)δ8.06(dd,J=8.4,1.1Hz,2H),7.79–7.74(m,1H),7.63(t,J=7.9Hz,2H),4.51(t,J=6. 1Hz, 2H), 4.28 (t, J = 6.1Hz, 2H), 2.43 (td, J = 7.3, 1.4Hz, 4H), 2.22 (p, J = 6.1Hz, 2H), 1.96 (p, J = 7.3Hz, 2H). 13 CNMR(125MHz, CDCl3)δ178.6,172.9,159.0,138.1,135.8,129.8,128.7,110.6,68.1,60.5,33.1,33.0,28.0,19.9.MS(ESI)m / z:437.0[M+Na] + .

[0101] 4-((4-((3-carboxypropionyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide (15a)

[0102] White solid, yield 60%, mp106-108℃. 1 HNMR (400MHz, CDCl3) δ8.11–8.04(m,2H),7.77(t,J=7.5Hz,1H),7.63(t,J=7.9Hz,2H),5.10(t,J=1.6Hz,2H),4.81–

[0103] 4.73(m,2H),2.74–2.65(m,4H). 13 CNMR(100MHz, CDCl3)δ177.2,171.4,158.1,138.1,135.9,129.9,128.8,110.8,83.9,78.8,58.8,52.3,28.8,28.8.HRMS(ESI)calcdforC 16 H 14 N2NaO9S433.0312[M+Na] + ,found433.0317.

[0104] 4-(4-((4-carboxybutyryl)oxy)but-2-yn-1-yloxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide (15b)

[0105] White solid, yield 76%, mp 94-96℃. 1HNMR (400MHz, CDCl3) δ8.07(d,J=7.6Hz,2H),7.77(t,J=7.5Hz,1H),7.63(t,J=7.8H z,2H),5.10(s,2H),4.74(s,2H),2.46(td,J=7.3,4.9Hz,4H),1.97(p,J=7.3Hz,2H). 13 CNMR(100MHz, CDCl3)δ179.0,172.1,158.0,137.7,135.8,129.8,128.7,110.6,83.9,78.6,58.6,52.0,32.8,19.6.HRMS(ESI)calcdforC 16 H 14 N2NaO9S447.0469[M+Na] + ,found447.0486.

[0106] (VI) Preparation of β-elemene nitric oxide donor derivatives 16a-f, 17a-f, 18a-f, 19a-f, 20a-f and 21a-f

[0107]

[0108] Scheme 4. Synthesis of β-elemene nitric oxide donor derivatives 16a-f, 17a-f, 18a-f, 19a-f, 20a-f, and 21a-f. Reagents and conditions: EDCI, DMAP, DCM, rt, 12 h.

[0109] As shown in Reaction Scheme 4, elemene intermediates 5-10 (0.2 mmol), NO donor intermediates 13a, 13b, 14a, 14b, 15a, and 15b (0.2 mmol), EDCI (0.2 mmol), and a catalytic amount of DMAP were stirred in anhydrous dichloromethane (3 mL) at room temperature for 12 h. The reaction was monitored by LC-MS. The reaction was quenched by adding water, and the reaction mixture was extracted with dichloromethane. The organic layer was washed sequentially with water and brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol volume ratio 100:1) to obtain a liquid product.

[0110] All intermediates used can be prepared according to the above (1) to (5), which will not be described in detail here.

[0111] Example 1: Preparation of Compound 1 (16a)

[0112]

[0113] Preparation of 4-(2-((4-(2-((2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-4-oxobutanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide.

[0114] To a solution of Intermediate 5 (0.2 mmol) in dichloromethane (3 mL) were added Intermediate 13a (0.2 mmol), EDCI (0.2 mmol), and DMAP (0.01 mmol), followed by stirring at room temperature for 12 h. The mixture was diluted with dichloromethane, washed sequentially with water and saturated water, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane:methanol, volume ratio, 100:1) to obtain a colorless, transparent liquid in a 65% yield.

[0115] 1 HNMR (500MHz, CDCl3) δ8.06–8.01(m,2H),7.77–7.71(m,1H),7.61(t,J=7.9Hz,2H),7.49–7.46(m,1H),7.38(d,J=2 .2Hz,1H),6.24(t,J=2.1Hz,1H),5.75(dd,J=17.4,10.9Hz,1H),5.00(d,J=16.7Hz,2H),4.88–4.81(m,2H),4.73(d ,J=15.3Hz,4H),4.62–4.58(m,2H),4.49(dt,J=8.7,4.5Hz,2H),4.19–4.09(m,J=6.1Hz,2H),3.11(d,J=13.5Hz,1H ),2.70–2.58(m,5H),2.44–2.37(m,2H),2.19–2.11(m,4H),1.92(t,J=10.9Hz,1H),1.60–1.37(m,6H),0.94(s,3H). 13 CNMR (125MHz, CDCl3) δ172.2,172.2,158.8,150.0,149.9,149.0,139.3,138.1,135.8,129.8,129.5,128.7,114.1,111.5,110.5,11 0.4,105.8,69.0,66.1,62.8,61.5,56.3,55.3,47.2,42.8,42.0,39.9,39.8,33.6,29.1,29.0,27.1,15.8.HRMS(ESI)m / z:calcdforC 35 H45 N5O9S[M+H] + ,712.3011;found,712.3034.

[0116] Example 2: Preparation of Compound 2 (16b)

[0117]

[0118] The preparation method of 4-(2-((5-(2-((2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-5-oxopentanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that succinic anhydride in (V) is replaced by glutaric anhydride.

[0119] In this example, a colorless transparent liquid was obtained with a yield of 73%.

[0120] 1 HNMR (500MHz, CDCl3) δ8.08–8.03(m,2H),7.76(t,J=7.5Hz,1H),7.62(t,J=7.8Hz,2H),7.50(d,J=1.7Hz,1H),7. 40(s,1H),6.26(t,J=1.8Hz,1H),5.77(dd,J=11.1,17.2Hz,1H),5.02(d,J=17.7Hz,2H),4.89–4.83(m,2H),4.74 (d,J=15.1Hz,4H),4.64–4.60(m,2H),4.52–4.48(m,2H),4.22–4.07(m,2H),3.11(d,J=13.4Hz,1H),2.63(s,2H) ,2.43(dt,J=7.3,23.3Hz,5H),2.15(s,3H),1.98(p,J=7.3Hz,3H),1.72(s,1H),1.61–1.39(m,6H),0.95(s,3H). 13CNMR (125MHz, CDCl3) δ172.8,172.7,158.7,149.9,149.9,148.9,139.3,138.1,135.7,129.8,129.5,128.7,114.0,111.5,110.5,110.4 ,105.8,69.0,66.0,62.4,61.2,56.3,55.4,47.2,42.7,42.0,39.9,39.8,33.6,33.2,33.1,27.0,20.0,15.8.HRMS(ESI)m / z:calcdforC 36 H 47 N5O9S[M+H] + ,726.3167;found,726.3158.

[0121] Example 3: Preparation of Compound 3 (16c)

[0122]

[0123] The preparation method of 4-(3-((4-(2-((2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-4-oxobutanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0124] In this example, a colorless transparent liquid was obtained with a yield of 67%.

[0125] 1HNMR(500MHz, CDCl3)δ8.03(d,J=7.7Hz,2H),7.73(t,J=7.5Hz,1H),7.60(t,J=7.8Hz,2H),7.49–7.46(m,1H),7.38(d, J=2.1Hz,1H),6.24(t,J=2.1Hz,1H),5.75(dd,J=10.8,17.5Hz,1H),5.00(d,J=16.0Hz,2H),4.87–4.82(m,2H),4.75–4 .69(m,4H),4.48(t,J=6.1Hz,2H),4.27(t,J=6.1Hz,2H),4.16–4.06(m,J=6.3Hz,2H),3.10(d,J=13.6Hz,1H),2.61(d, J=29.4Hz,6H),2.40(dt,J=5.4,12.3Hz,1H),2.21–2.12(m,6H),1.91(t,J=9.6Hz,1H),1.60–1.37(m,6H),0.93(s,3H). 13 CNMR (125MHz, CDCl3) δ172.2,158.9,149.9,149.8,148.9,139.2,138.0,135.7,129.7,129.4,128.6,114.0,111.4,110.5,110.3,10 5.7,68.0,66.0,62.6,60.5,56.2,55.2,47.1,42.7,41.9,39.8,39.7,33.5,29.1,29.0,27.9,27.0,15.8.HRMS(ESI)m / z:calcdforC 36 H 47 N5O9S[M+H] + ,726.3167;found,726.3157.

[0126] Example 4: Preparation of Compound 4 (16d)

[0127]

[0128] The preparation method of 4-(3-((5-(2-((2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-5-oxopentanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 2, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0129] In this example, a colorless transparent liquid was obtained with a yield of 62%.

[0130] 1 HNMR(500MHz, CDCl3)δ8.05(dd,J=8.5,1.1Hz,2H),7.77–7.73(m,1H),7.64–7.60(m,2H),7.50(d,J=1.5Hz,1H),7.39(d, J=1.9Hz,1H),6.26(t,J=2.0Hz,1H),5.77(dd,J=17.5,10.8Hz,1H),5.02(d,J=18.7Hz,2H),4.89–4.83(m,2H),4.74(d,J =13.5Hz,4H),4.50(t,J=6.1Hz,2H),4.26(t,J=6.1Hz,2H),4.20–4.08(m,2H),3.18–3.06(m,1H),2.62(d,J=12.5Hz,2H) ,2.38(dt,J=11.6,7.4Hz,5H),2.23–2.12(m,6H),1.96–1.93(m,2H),1.82–1.67(m,1H),1.60–1.38(m,6H),0.95(s,3H). 13 CNMR (125MHz, CDCl3) δ172.8,172.8,158.9,149.9,149.9,148.9,139.2,138.1,135.7,129.8,129.5,128.6,114.0,111.5,110.5,110.4,1 05.8,68.0,66.0,62.4,60.3,56.3,55.4,47.2,42.7,42.0,39.8,39.8,33.6,33.3,33.2,28.0,27.0,20.1,15.8.HRMS(ESI)m / z:calcdforC 37 H 49 N5O9S[M+H] + ,740.3324;found,740.3329.

[0131] Example 5: Preparation of Compound 5 (16e)

[0132]

[0133] The preparation method of 4-((4-((4-(2-((2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-4-oxobutanoyl)oxy)but-2-ynyl-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0134] In this example, a colorless transparent liquid was obtained with a yield of 60%.

[0135] 1 HNMR(500MHz, CDCl3)δ8.07(dd,J=8.5,1.2Hz,2H),7.79–7.74(m,1H),7.66–7.60(m,2H),7.50(d,J=1 .5Hz,1H),7.43–7.36(m,1H),6.26(t,J=2.1Hz,1H),5.77(dd,J=17.4,10.9Hz,1H),5.11–4.99(m,4H), 4.90–4.84(m,2H),4.79–4.72(m,6H),4.24–4.09(m,2H),3.12(d,J=13.2Hz,1H),2.66(tt,J=9.3,4.7 Hz, 6H), 2.48–2.38 (m, 1H), 2.18 (d, J = 19.3Hz, 4H), 1.98–1.88 (m, 1H), 1.63–1.41 (m, 6H), 0.96 (s, 3H). 13 CNMR (125MHz, CDCl3) δ172.1,171.5,158.0,149.9,149.9,149.0,139.3,137.9,135.8,129.8,129.5,128.7,114.1,111.5,110.6,110.4,1 05.8,83.9,78.7,66.0,62.7,58.7,56.3,55.3,52.2,47.2,42.7,42.0,39.9,39.8,33.6,29.0,28.9,27.0,15.8.HRMS(ESI)m / z:calcdfor C 37 H 45 N5O9S[M+H] + ,736.3011;found,736.3013.

[0136] Example 6: Preparation of Compound 6 (16f)

[0137]

[0138] The preparation method of 4-((4-((5-(2-((2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-5-oxopentanoyl)oxy)but-2-ynyl-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 2, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0139] In this example, a colorless transparent liquid was obtained with a yield of 87%.

[0140] 1 HNMR (500MHz, CDCl3) δ8.06–8.03(m,2H),7.77–7.72(m,1H),7.61(t,J=7.9Hz,2H),7.48(d,J=1.7Hz,1H),7.38(d, J=2.1Hz,1H),6.24(t,J=2.0Hz,1H),5.75(dd,J=10.8,17.5Hz,1H),5.07(t,J=1.5Hz,2H),5.00(d,J=16.6Hz,2H),4 .87–4.82(m,2H),4.75–4.71(m,6H),4.13(tt,J=6.3,14.0Hz,2H),3.11(d,J=13.6Hz,1H),2.67–2.57(m,2H),2.40( dt,J=7.3,24.0Hz,5H),2.16(d,J=15.9Hz,4H),1.94(td,J=5.7,11.5,13.1Hz,3H),1.61–1.37(m,6H),0.94(s,3H). 13 CNMR (125MHz, CDCl3) δ172.8,172.1,158.0,149.9,149.8,148.9,139.2,137.9,135.8,129.8,129.4,128.7,114.0,111.5,110.6,110.4,105. 8,84.0,78.6,66.0,62.4,58.7,56.3,55.4,51.9,47.2,42.7,42.0,39.8,39.7,33.6,33.2,32.9,27.0,20.0,15.8.HRMS(ESI)m / z:calcdforC 38 H 47 N5O9S[M+H] +,750.3167;found,750.3174.

[0141] Example 7: Preparation of Compound 7 (17a)

[0142]

[0143] The preparation method of 4-(2-((4-((1-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-4-oxobutanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that N-methyl-2-hydroxyethylamine in (III) is replaced by 4-hydroxymethylpiperidine.

[0144] In this example, a colorless transparent liquid was obtained with a yield of 61%.

[0145] 1 HNMR(500MHz, CDCl3)δ8.05(dd,J=8.5,1.2Hz,2H),7.76–7.72(m,1H),7.61(td, J=7.6,1.7Hz,2H),7.49(d,J=1.8Hz,1H),7.38(d,J=1.9Hz,1H),6.25(t,J=2.1Hz ,1H),5.75(dd,J=17.5,10.8Hz,1H),5.00(d,J=6.8Hz,2H),4.87–4.81(m,2H),4 .76–4.70(m,4H),4.61(dd,J=5.4,3.7Hz,2H),4.50(dd,J=5.9,3.1Hz,2H),3.97–

[0146] 3.89(m,2H),2.98(d,J=13.8Hz,1H),2.77(d,J=9.5Hz,2H),2.69–2.64(m,4H),2.59(d,J=13.9 Hz, 1H), 2.14 (dd, J=12.5, 3.2Hz, 1H), 1.91 (q, J=11.5Hz, 2H), 1.73–1.29 (m, 12H), 0.95 (s, 3H). 13CNMR (125MHz, CDCl3) δ172.2,172.1,158.8,149.9,149.9,148.5,139.3,138.1,135.7,129.8,129.4,128.7,113.5,111.5,110.5,110.3,1 05.8,69.3,68.9,66.5,61.5,56.4,54.1,52.5,47.9,41.9,39.8,39.8,35.5,33.6,29.0,29.0,28.9,27.1,15.9.HRMS(ESI)m / z:calcdforC 38 H 49 N5O9S[M+H] + ,752.3324;found,752.3317.

[0147] Example 8: Preparation of Compound 8 (17b)

[0148]

[0149] The preparation method of 4-(2-((5-((1-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl)-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-5-oxopentanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 7, except that succinic anhydride in (V) is replaced by glutaric anhydride.

[0150] In this example, a colorless transparent liquid was obtained with a yield of 58%.

[0151] 1HNMR(500MHz, CDCl3)δ8.05(dd,J=1.1,8.4Hz,2H),7.78–7.72(m,1H),7.64–7.60(m,2H),7.50(d,J=1.7Hz,1H),7.38(d,J=2.1Hz ,1H),6.26(t,J=2.0Hz,1H),5.76(dd,J=10.8,17.5Hz,1H),5.00(d,J=6.4Hz,2H),4.88–4.82(m,2H),4.75(d,J=4.8Hz,4H),4.62 (dd,J=3.7,5.4Hz,2H),4.52–4.48(m,2H),3.93(d,J=6.2Hz,2H),2.98(d,J=12.7Hz,1H),2.82–2.72(m,2H),2.59(d,J=13.3Hz,1 H),2.42(dt,J=7.3,22.1Hz,4H),2.15(d,J=11.2Hz,1H),2.04–1.84(m,5H),1.75–1.66(m,1H),1.67–1.38(m,10H),0.95(s,3H). 13 CNMR (125MHz, CDCl3) δ173.0,172.7,158.8,150.0,149.9,148.6,139.3,138.1,135.8,129.8,129.5,128.7,113.5,111.5,110.5,110.3,105. 8,69.1,69.0,66.5,61.2,56.4,54.2,52.5,47.9,42.0,39.9,39.8,35.5,33.7,33.3,33.2,29.1,27.1,20.1,16.0.HRMS(ESI)m / z:calcdforC 39 H 51 N5O9S[M+H] + ,766.348;found,766.3478.

[0152] Example 9: Preparation of Compound 9 (17c)

[0153]

[0154] The preparation method of 4-(3-((4-((1-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-4-oxobutanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 7, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0155] In this example, a colorless transparent liquid was obtained with a yield of 68%.

[0156] 1 HNMR (500MHz, CDCl3) δ8.08–8.01(m,2H),7.75(t,J=7.5Hz,1H),7.62(t,J=7.8Hz,2H),7.52–7.46(m,1H) ,7.38(s,1H),6.26(t,J=1.9Hz,1H),5.77(dd,J=10.8,17.4Hz,1H),5.00(s,2H),4.89–4.81(m,2H),4.75( d,J=4.1Hz,4H),4.50(t,J=6.1Hz,2H),4.30(t,J=6.1Hz,2H),3.90(d,J=6.1Hz,2H),2.97(d,J=12.9Hz,1H ),2.82–2.72(m,2H),2.63(s,5H),2.25–2.10(m,3H),2.04–1.75(m,3H),1.75–1.35(m,11H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.4,172.3,159.0,150.0,149.9,148.6,139.3,

[0157] 138.1,135.7,129.8,129.5,128.7,113.5,111.5,110.6,110.3,105.8,69.3,68.0,66.5,60.6,56.4,5 4.1,52.5,47.9,42.0,39.9,39.8,35.5,33.7,29.1,29.0,28.0,27.1,16.0.HRMS(ESI)m / z:calcdforC 39 H 51 N5O9S[M+H] + ,766.348;found,766.3496.

[0158] Example 10: Preparation of Compound 10 (17d)

[0159]

[0160] The preparation method of 4-(3-((5-((1-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-5-oxopentanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 8, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0161] In this example, a colorless transparent liquid was obtained with a yield of 56%.

[0162] 1 HNMR(500MHz, CDCl3)δ8.04(dd,J=1.1,8.4Hz,2H),7.77–7.72(m,1H),7.65–7.59(m,2H),7.49(d,J=1.7Hz,1H),7.38(d,J=2 .1Hz,1H),6.25(t,J=2.0Hz,1H),5.76(dd,J=10.8,17.5Hz,1H),5.00(d,J=7.5Hz,2H),4.88–4.81(m,2H),4.74(d,J=3.9Hz,4 H),4.49(t,J=6.1Hz,2H),4.26(t,J=6.1Hz,2H),3.95–3.89(m,2H),2.98(d,J=13.4Hz,1H),2.83–2.72(m,2H),2.59(d,J=13 .6Hz,1H),2.38(dt,J=7.4,10.7Hz,4H),2.23–2.12(m,3H),2.05–1.80(m,5H),1.68(s,1H),1.67–1.36(m,10H),0.95(s,3H). 13 CNMR (125MHz, CDCl3) δ173.0,172.8,158.9,149.9,149.9,148.5,139.3,138.1,135.8,129.8,129.4,128.6,113.4,111.5,110.5,110.3,105.8, 69.0,68.0,66.5,60.3,56.4,54.1,52.5,47.9,41.9,39.8,39.8,35.5,33.7,33.3,33.2,29.0,28.0,27.1,20.2,16.0.HRMS(ESI)m / z:calcdforC 40 H 53N5O9S[M+H] + ,780.3637;found,780.3634.

[0163] Example 11: Preparation of Compound 11 (17e)

[0164]

[0165] The preparation method of 4-((4-((4-((1-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-4-oxobutanoyl)oxy)but-2-ynyl-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 7, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0166] In this example, a colorless transparent liquid was obtained with a yield of 49%.

[0167] 1 HNMR(500MHz, CDCl3)δ8.06(dd,J=1.2,8.5Hz,2H),7.79–7.72(m,1H),7.65–7.59(m,2H),7.49(d,J=1.4Hz,1H),7.38(d ,J=2.1Hz,1H),6.25(t,J=2.1Hz,1H),5.76(dd,J=10.8,17.5Hz,1H),5.08(t,J=1.7Hz,2H),5.00(d,J=8.2Hz,2H),4.87–

[0168] 4.81(m,2H),4.74(d,J=4.6Hz,6H),3.95(d,J=5.9Hz,2H),2.99(d,J=13.6Hz,1H),2.78(d,J=8.4Hz, 2H),2.71–2.56(m,5H),2.15(d,J=10.4Hz,1H),1.96–1.85(m,2H),1.72–1.31(m,12H),0.95(s,3H). 13CNMR (125MHz, CDCl3) δ172.1,171.6,158.0,150.0,149.9,148.5,139.3,137.9,135.8,129.8,129.4,128.7,113.5,111.5,110.7,110.3,105. 8,83.9,78.7,69.4,66.5,58.7,56.4,54.1,52.5,52.2,47.9,41.9,39.8,39.8,35.5,33.7,29.0,29.0,27.1,16.0.HRMS(ESI)m / z:calcdforC 40 H 49 N5O9S[M+H] + ,776.3324;found,776.3298.

[0169] Example 12: Preparation of Compound 12 (17f)

[0170]

[0171] The preparation method of 4-((4-((5-((1-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-5-oxopentanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 8, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0172] In this example, a colorless transparent liquid was obtained with a yield of 25%.

[0173] 1HNMR (500MHz, CDCl3) δ8.09–8.04(m,2H),7.79–7.74(m,1H),7.63(t,J=7.9Hz,2H),7.50(d,J=1.5Hz,1H),7.40–7.37(m ,1H),6.26(t,J=2.0Hz,1H),5.77(dd,J=10.8,17.4Hz,1H),5.09(t,J=1.6Hz,2H),5.00(s,2H),4.88–4.82(m,2H),4.77 –4.72(m,6H),3.94(d,J=6.0Hz,2H),2.98(d,J=13.1Hz,1H),2.83–2.73(m,2H),2.60(d,J=13.5Hz,1H),2.41(dt,J=7.4 ,21.6Hz,4H),2.15(d,J=11.0Hz,1H),1.95(dp,J=8.4,9.4,19.4Hz,5H),1.68(s,1H),1.66–1.37(m,10H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.9,172.2,158.0,150.0,149.9,148.6,139.3,138.0,135.8,129.8,129.5,128.8,113.4,111.5,110.7,110.3,105.8, 84.1,78.6,69.1,66.5,58.7,56.4,54.2,52.5,52.0,47.9,42.0,39.8,35.5,33.7,33.2,33.1,29.1,27.1,20.1,16.0.HRMS(ESI)m / z:calcdfor C 41 H 51 N5O9S[M+H] + ,790.348;found,790.3474.

[0174] Example 13: Preparation of Compound 13 (18a)

[0175]

[0176] The preparation method of 4-(2-((4-(2-(4-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-4-oxobutanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that N-methyl-2-hydroxyethylamine in (III) is replaced by 1-(2-hydroxyethyl)piperazine.

[0177] In this example, a colorless transparent liquid was obtained with a yield of 54%.

[0178] 1 HNMR (500MHz, CDCl3) δ8.11–8.02(m,2H),7.76(t,J=7.5Hz,1H),7.63(t,J=7.9Hz,2H),7.50(s,1H),7.39(s,1H),6. 28–6.23(m,1H),5.77(dd,J=10.8,17.1Hz,1H),5.02(d,J=17.6Hz,2H),4.85(dd,J=6.6,13.6Hz,2H),4.75(s,4H),4. 65–4.59(m,2H),4.54–4.48(m,2H),4.23(t,J=5.4Hz,2H),3.08–2.96(m,1H),2.67(d,J=16.7Hz,8H),2.57–2.48(m,3 H),2.42(d,J=19.9Hz,2H),2.20–2.10(m,1H),1.93–1.86(m,1H),1.85–1.68(m,2H),1.61–1.40(m,6H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.2,172.2,158.8,149.9,149.9,147.9,139.3,138.1,135.8,129.8,129.5,128.8,113.9,111.6,110.5,110.4 ,105.9,69.0,66.2,62.4,61.5,56.7,56.4,53.6,53.0,47.9,42.0,39.9,39.9,33.7,29.1,29.0,27.1,16.0.HRMS(ESI)m / z:calcdforC 38 H 50 N6O9S[M+H] + ,767.3433;found,767.343.

[0179] Example 14: Preparation of Compound 14 (18b)

[0180]

[0181] The preparation method of 4-(2-((5-(2-(4-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-5-oxopentanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 13, except that succinic anhydride in (V) is replaced by glutaric anhydride.

[0182] In this example, a colorless transparent liquid was obtained with a yield of 45%.

[0183] 1 HNMR(500MHz, CDCl3)δ8.06(dd,J=1.2,8.5Hz,2H),7.79–7.74(m,1H),7.62(td,J=1.7,7.6Hz,2H),7.50(d,J=1.4Hz,1H),7.39(d,J=1.9Hz, 1H),6.26(t,J=2.1Hz,1H),5.77(dd,J=10.8,17.4Hz,1H),5.02(d,J=13.7Hz,2H),4.89–4.82(m,2H),4.76(s,4H),4.62(dd,J=3.7,5.4Hz,2H ),4.50(dd,J=3.7,5.4Hz,2H),4.21(t,J=5.9Hz,2H),3.07–2.94(m,1H),2.64(t,J=5.6Hz,3H),2.56–2.50(m,2H),2.44(dt,J=7.3,23.1Hz, 7H),2.30(d,J=16.2Hz,1H),2.19–2.12(m,1H),1.98(p,J=7.3Hz,2H), 1.94–1.86(m,1H),1.83–1.66(m,2H),1.62–1.40(m,6H),0.96(s,3H). 13CNMR (125MHz, CDCl3) δ172.9,172.7,158.8,149.9,149.9,148.1,139.3,138.1,135.8,129.8,129.4,128.7,113.9,111.5,110.5,110.3,1 05.8,69.0,66.2,62.0,61.2,56.7,56.4,53.5,53.0,47.9,41.9,39.8,39.8,33.6,33.2,33.1,27.1,20.0,15.9.HRMS(ESI)m / z:calcdforC 39 H 52 N6O9S[M+H] + ,781.3589;found,781.3541.

[0184] Example 15: Preparation of Compound 15 (18c)

[0185]

[0186] The preparation method of 4-(3-((4-(2-(4-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-4-oxobutanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 13, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0187] In this example, a light yellow liquid was obtained with a yield of 80%.

[0188] 1HNMR(500MHz, CDCl3)δ8.07–8.04(m,2H),7.78–7.73(m,1H),7.65–7.60(m,2H),7.52–7.49(m,1H),7.40(d,J =2.0Hz,1H),6.27(t,J=2.0Hz,1H),5.77(dd,J=10.8,17.5Hz,1H),5.03(d,J=24.5Hz,2H),4.89–4.82(m,2H) ,4.81–4.72(m,4H),4.51(t,J=6.1Hz,2H),4.30(t,J=6.1Hz,2H),4.20(t,J=5.8Hz,2H),3.06(d,J=15.5Hz,1 H),2.68–2.41(m,14H),2.21(q,J=6.1Hz,4H),1.91(dd,J=7.9,18.0Hz,1H),1.60–1.40(m,6H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.3,158.9,149.9,149.8,147.9,139.2,138.0,135.7,129.8,129.4,128.6,114.1,111.5,110.5,110.4,105. 8,68.0,66.1,62.1,60.6,56.5,56.4,53.3,52.8,47.9,41.9,39.8,39.8,33.6,29.1,29.0,28.0,27.0,15.9.HRMS(ESI)m / z:calcdforC 39 H 52 N6O9S[M+H] + ,781.3589;found,781.357.

[0189] Example 16: Preparation of Compound 16 (18d)

[0190]

[0191] The preparation method of 4-(3-((5-(2-(4-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-5-oxopentanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 14, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0192] In this example, a colorless transparent liquid was obtained with a yield of 68%.

[0193] 1 HNMR(500MHz, CDCl3)δ8.06(d,J=8.0Hz,2H),7.76(t,J=7.5Hz,1H),7.63(t,J=7.9Hz,2H),7.51(s,1H),7.40(s,1H), 6.27(s,1H),5.77(dd,J=10.8,16.6Hz,1H),5.02(d,J=13.6Hz,2H),4.89–4.82(m,2H),4.76(s,4H),4.50(t,J=6.1Hz ,2H),4.24(dt,J=5.8,27.0Hz,4H),3.01(d,J=21.1Hz,1H),2.65(s,3H),2.57–2.50(m,2H),2.43–2.36(m,5H),2.35– 2.27(m,1H),2.21(p,J=6.1Hz,2H),2.18–2.10(m,1H),1.94(tt,J=7.7,13.8Hz,3H),1.73–1.39(m,10H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.9,158.9,149.9,149.9,148.1,139.3,138.1,135.8,129.8,129.5,128.6,113.9,111.6,110.5,110.3,105.8,6 8.1,66.2,62.0,60.4,56.8,56.4,53.6,53.0,47.9,42.0,39.9,39.8,33.7,33.3,33.2,28.0,27.1,20.2,15.9.HRMS(ESI)m / z:calcdforC 40 H 54 N6O9S[M+H] + ,795.3746;found,795.3701.

[0194] Example 17: Preparation of Compound 17 (18e)

[0195]

[0196] The preparation method of 4-((4-((4-(2-(4-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-))yl)-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-4-oxobutanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 13, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0197] In this example, a colorless transparent liquid was obtained with a yield of 45%.

[0198] 1 HNMR (500MHz, CDCl3) δ8.08(d,J=8.0Hz,2H),7.77(t,J=7.5Hz,1H),7.63(t,J=7.7Hz,2H),7.51(s,1H),7. 39(s,1H),6.27(s,1H),5.77(dd,J=10.7,17.4Hz,1H),5.12–4.99(m,4H),4.88–4.73(m,8H),4.24(t,J=5. 7Hz,2H),3.08–2.97(m,1H),2.66(d,J=12.3Hz,7H),2.56–2.50(m,2H),2.43(dt,J=7.4,15.7Hz,2H),2.35 –2.28(m,1H),2.19–2.12(m,1H),1.90(d,J=7.3Hz,1H),1.75–1.66(m,3H),1.61–1.41(m,6H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.1,171.6,158.0,149.9,149.9,148.1,139.3,137.9,135.8,129.8,129.4,128.7,113.9,111.6,110.7,110.3,105. 8,83.9,78.8,66.2,62.4,58.7,56.7,56.4,53.5,53.0,52.2,47.9,41.9,39.8,39.8,33.6,29.0,28.9,27.1,15.9.HRMS(ESI)m / z:calcdforC 40 H 50 N6O9S[M+H] + ,791.3433;found,791.3423.

[0199] Example 18: Preparation of Compound 18 (18f)

[0200]

[0201] The preparation method of 4-((4-((5-(2-(4-(2-((1R,2S,5R)-5-(3-(1H-pyrazol-1-yl)prop-1-en-2-yl)-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-5-oxopentanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 14, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0202] In this example, a colorless transparent liquid was obtained with a yield of 56%.

[0203] 1 HNMR(500MHz, CDCl3)δ8.07(d,J=7.8Hz,2H),7.77(t,J=7.5Hz,1H),7.63(t,J= 7.8Hz,2H),7.53–7.49(m,1H),7.40(s,1H),6.27(t,J=1.8Hz,1H),5.77(dd,J=

[0204] 11.3,17.3Hz,1H),5.05(d,J=45.6Hz,4H),4.89–4.83(m,2H),4.80–4.72(m ,6H),4.23(t,J=5.4Hz,2H),3.09–2.95(m,1H),2.65(s,3H),2.55(d,J=14. 3Hz,3H),2.43(dt,J=7.3,22.7Hz,6H),2.34–2.27(m,1H),2.22–2.12(m,1H ),1.97(p,J=7.3Hz,3H),1.77–1.67(m,2H),1.60–1.41(m,6H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.8,172.2,158.0,149.9,149.9,148.1,139.3,137.9,135.8,129.8,129.4,128.7,114.0,111.6,

[0205] 110.7,110.4,105.8,84.1,78.6,66.2,61.9,58.7,56.7,56.4,53.5,53.0,51.9,47 .9,41.9,39.8,39.8,33.6,33.2,33.0,27.1,20.0,15.9.HRMS(ESI)m / z:calcdforC 41 H 52 N6O9S[M+H] + ,805.3589;found,805.3593.

[0206] Example 19: Preparation of Compound 19 (19a)

[0207]

[0208] The preparation method of 4-(2-((4-(2-((2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-4-oxobutanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that the pyrazole in (II) is replaced by N-ethylpiperazine.

[0209] In this example, a colorless transparent liquid was obtained with a yield of 49%.

[0210] 1 HNMR(500MHz, CDCl3)δ8.09–8.03(m,2H),7.75(tt,J=1.2,7.2Hz,1H),7.66–7.59(m,2H),5.79(dd,J=10.8 ,17.5Hz,1H),5.03(s,1H),4.94–4.83(m,4H),4.77(s,1H),4.64–4.60(m,2H),4.53–4.49(m,2H),4.17(td, J=3.3,6.0Hz,2H),3.13(d,J=13.7Hz,1H),2.95–2.88(m,2H),2.71–2.61(m,7H),2.60–2.27(m,10H),2.23 –2.16(m,4H),2.12–2.02(m,1H),1.66–1.57(m,2H),1.51–1.37(m,4H),1.10(t,J=7.2Hz,3H),0.97(s,3H). 13CNMR (125MHz, CDCl3) δ172.2,172.2,158.8,150.9,150.3,149.3,138.2,135.8,129.8,128.8,113.9,110.9,110.5,110.2,69.0,66.1 ,63.5,62.9,61.5,55.4,53.0,52.9,52.4,47.6,42.9,42.5,40.1,40.0,34.0,29.1,29.0,27.3,15.9,11.9.HRMS(ESI)m / z:calcdforC 38 H 55 N5O9S[M+H] + ,758.3793;found,758.3749.

[0211] Example 20: Preparation of Compound 20 (19b)

[0212]

[0213] The preparation method of 4-(2-((5-(2-((2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-5-oxopentanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 19, except that succinic anhydride in (V) is replaced by glutaric anhydride.

[0214] In this example, a colorless transparent liquid was obtained with a yield of 58%.

[0215] 1HNMR(500MHz, CDCl3)δ8.04(dd,J=1.1,8.5Hz,2H),7.77–7.72(m,1H),7.64–7.58(m,2H),5.78(dd,J=10.8,17.5H z,1H),5.03(s,1H),4.94–4.82(m,4H),4.76(s,1H),4.61(dd,J=3.8,5.4Hz,2H),4.49(dd,J=3.2,5.9Hz,2H),4.1 5(td,J=3.5,6.0Hz,2H),3.12(d,J=13.7Hz,1H),2.95–2.88(m,2H),2.71–2.34(m,17H),2.22–2.15(m,4H),2.07( d,J=7.8Hz,1H),1.97(p,J=7.4Hz,2H),1.65–1.56(m,2H),1.50–1.36(m,4H),1.09(t,J=7.2Hz,3H),0.96(s,3H). 13 CNMR (125MHz, CDCl3) δ172.9,172.7,158.8,150.9,150.3,149.3,138.1,135.8,129.8,128.7,113.8,110.9,110.5,110.2,69.0,66.1,63 .5,62.5,61.2,55.5,53.0,52.9,52.4,47.5,42.9,42.5,40.1,39.9,34.0,33.2,33.1,27.3,20.0,15.9,11.9.HRMS(ESI)m / z:calcdforC 39 H 57 N5O9S[M+H] + ,772.395;found,772.3926.

[0216] Example 21: Preparation of Compound 21 (19c)

[0217]

[0218] The preparation method of 4-(3-((4-(2-((2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-4-oxobutanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 19, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0219] In this example, a colorless transparent liquid was obtained with a yield of 42%.

[0220] 1 HNMR(500MHz, CDCl3)δ8.06(dd,J=1.1,8.4Hz,2H),7.80–7.72(m,1H),7.66–7.60(m,2H),5.80(dd,J=10.8,17.5H z,1H),5.04(s,1H),4.96–4.84(m,4H),4.78(s,1H),4.51(t,J=6.1Hz,2H),4.30(t,J=6.1Hz,2H),4.14(td,J=3.3, 6.0Hz,2H),3.14(d,J=13.7Hz,1H),2.99–2.88(m,2H),2.71–2.61(m,7H),2.59–2.41(m,8H),2.24–2.16(m,6H),2. 13–2.01(m,2H),2.02–1.91(m,1H),1.61(q,J=12.9Hz,2H),1.52–1.39(m,4H),1.14(t,J=7.2Hz,3H),0.98(s,3H). 13 CNMR (125MHz, CDCl3) δ172.3,172.3,159.0,150.9,150.3,149.3,138.1,135.8,129.8,128.7,113.9,110.9,110.6,110.2,68.0,66.1 ,63.4,62.8,60.6,55.4,52.9,52.4,47.5,42.8,42.5,40.1,40.0,34.0,29.1,29.1,28.0,27.3,15.9,11.8.HRMS(ESI)m / z:calcdforC 39 H 57 N5O9S[M+H] + ,772.395;found,772.3937.

[0221] Example 22: Preparation of Compound 22 (19d)

[0222]

[0223] The preparation method of 4-(3-((5-(2-((2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-5-oxopentanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 20, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0224] In this example, a colorless transparent liquid was obtained with a yield of 43%.

[0225] 1 HNMR(500MHz, CDCl3)δ8.05(dd,J=1.1,8.4Hz,2H),7.78–7.72(m,1H),7.66–7.60(m,2H),5.79(dd,J=10.8,17.5Hz,1 H),5.04(s,1H),4.95–4.84(m,4H),4.77(s,1H),4.50(t,J=6.1Hz,2H),4.27(t,J=6.1Hz,2H),4.15(tt,J=4.0,7.7Hz, 2H),3.14(d,J=13.6Hz,1H),2.98–2.89(m,2H),2.66(td,J=7.0,13.5,14.0Hz,3H),2.60–2.34(m,13H),2.23–2.16(m, 6H), 2.01 (dp, J=7.4, 8.1, 62.7Hz, 4H), 1.61 (q, J=12.8Hz, 2H), 1.51–1.38 (m, 4H), 1.15 (t, J=7.2Hz, 3H), 0.98 (s, 3H). 13 CNMR(125MHz, CDCl3)δ172.9,172.9,158.9,150.9,150.3,149.3,138.1,135.8,129.8,128.7,113.9,110.9,110.6,110.2,68.1,66.1,63.5 ,62.5,60.4,55.5,52.9,52.9,52.4,47.5,42.9,42.5,40.1,40.0,34.0,33.3,33.2,28.1,27.3,20.2,15.9,11.8.HRMS(ESI)m / z:calcdforC 40 H 59 N5O9S[M+H] + ,786.4106;found,786.4089.

[0226] Example 23: Preparation of Compound 23 (19e)

[0227]

[0228] The preparation method of 4-((4-((4-(2-((2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-4-oxobutanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 19, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0229] In this example, a colorless transparent liquid was obtained with a yield of 32%.

[0230] 1 HNMR (500MHz, CDCl3) δ8.09–8.04(m,2H),7.78–7.74(m,1H),7.63(td,J=1.7,7.6Hz,2H),5.79(dd,J =10.8,17.5Hz,1H),5.09(t,J=1.7Hz,2H),5.03(s,1H),4.94–4.84(m,4H),4.79–4.74(m,3H),4.17(t t,J=4.1,7.7Hz,2H),3.13(d,J=13.6Hz,1H),2.97–2.88(m,2H),2.72–2.36(m,17H),2.23–2.15(m,4 H),2.10–2.02(m,1H),1.61(q,J=12.9Hz,2H),1.51–1.39(m,4H),1.11(t,J=7.2Hz,3H),0.97(s,3H). 13 CNMR (125MHz, CDCl3) δ172.1,171.6,158.0,151.0,150.3,149.3,138.0,135.8,129.8,128.8,113.9,110.8,110.7,110.2,83.9,78.8,66 .1,63.5,62.9,58.7,55.4,53.1,53.0,52.4,52.2,47.5,42.8,42.5,40.1,40.0,34.0,29.0,28.9,27.3,15.9,11.9.HRMS(ESI)m / z:calcd forC 40 H 55 N5O9S[M+H] +,782.3793;found,782.3796.

[0231] Example 24: Preparation of Compound 24 (19f)

[0232]

[0233] The preparation method of 4-((4-((5-(2-((2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)(methyl)amino)ethoxy)-5-oxopentanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 20, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0234] In this example, a colorless transparent liquid was obtained with a yield of 53%.

[0235] 1 HNMR(500MHz, CDCl3)δ8.04(dd,J=1.1,8.4Hz,2H),7.77–7.73(m,1H),7.64–7.59(m,2H),5.79(dd,J=10.8,17.5Hz,1 H),5.03(s,1H),4.94–4.82(m,4H),4.77(s,1H),4.49(t,J=6.1Hz,2H),4.26(t,J=6.1Hz,2H),4.18–4.10(m,2H),3.1 3(d,J=13.7Hz,1H),2.96–2.86(m,2H),2.65(td,J=7.2,13.5,14.1Hz,3H),2.51–2.35(m,12H),2.22–2.15(m,6H),2. 07(t,J=11.5Hz,1H),1.94(p,J=7.4Hz,2H),1.66–1.56(m,2H),1.50–1.37(m,4H),1.09(t,J=7.2Hz,3H),0.97(s,3H). 13CNMR (125MHz, CDCl3) δ172.8,172.1,158.0,151.0,150.3,149.3,138.0,135.8,129.8,128.8,113.8,110.8,110.7,110.2,84.1,78.6,66.1,63 .5,62.5,58.7,55.5,53.1,53.0,52.4,51.9,47.5,42.9,42.5,40.1,39.9,33.9,33.2,33.0,27.3,20.0,15.9,11.9.HRMS(ESI)m / z:calcdforC 41 H 57 N5O9S[M+H] + ,796.395;found,796.3958.

[0236] Example 25: Preparation of Compound 25 (20a)

[0237]

[0238] The preparation method of 4-(2-((4-((1-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-4-oxobutanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that N-methyl-2-hydroxyethylamine in (III) is replaced by 4-hydroxymethylpiperidine.

[0239] In this example, a colorless transparent liquid was obtained with a yield of 50%.

[0240] 1HNMR (500MHz, CDCl3) δ8.08–8.03(m,2H),7.75(t,J=7.5Hz,1H),7.62(t,J=7.9Hz,2H),5.79(dd,J=10.8,17.5Hz,1H),5.01( s,1H),4.93–4.82(m,4H),4.76(d,J=2.1Hz,1H),4.64–4.59(m,2H),4.54–4.48(m,2H),3.93(d,J=6.2Hz,2H),2.94(dt,J=13 .6,41.4Hz,4H),2.81(t,J=11.0Hz,2H),2.69–2.59(m,6H),2.55–2.36(m,8H),2.18(dd,J=3.1,12.7Hz,1H),2.06(t,J=11.5 Hz,1H),1.96–1.90(m,1H),1.70(d,J=11.6Hz,1H),1.65–1.55(m,5H),1.52–1.30(m,6H),1.08(t,J=7.2Hz,3H),0.98(s,3H). 13 CNMR (125MHz, CDCl3) δ172.3,172.2,158.8,151.0,150.3,148.8,138.1,135.8,129.8,128.7,113.4,110.8,110.5,110.1,69.4,69.0,66.7 ,63.6,61.5,54.2,53.1,52.9,52.6,52.4,48.3,42.4,40.1,40.0,35.5,34.1,29.1,29.0,29.0,27.3,16.1,11.9.HRMS(ESI)m / z:calcdforC 41 H 59 N5O9S[M+H] + ,798.4106;found,798.4096.

[0241] Example 26: Preparation of Compound 26 (20b)

[0242]

[0243] The preparation method of 4-(2-((5-((1-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-5-oxopentanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 25, except that succinic anhydride in (V) is replaced by glutaric anhydride.

[0244] In this example, a colorless transparent liquid was obtained with a yield of 29%.

[0245] 1 HNMR(500MHz, CDCl3) δ8.06(d,J=7.5Hz,2H),7.76(t,J=7.5Hz,1H),7.63(t,J=7.9Hz,2H),5.80(dd,J=10.8,17.5Hz, 1H),5.03(s,1H),4.94–4.84(m,4H),4.78(s,1H),4.65–4.59(m,2H),4.53–4.48(m,2H),3.93(d,J=6.3Hz,2H),3.04– 2.88(m,4H),2.86–2.80(m,2H),2.64(d,J=13.9Hz,2H),2.55–2.38(m,12H),2.20(d,J=12.6Hz,1H),2.07(s,1H),1.9 8(dt,J=7.3,14.6Hz,4H),1.72(s,1H),1.61(d,J=12.3Hz,4H),1.56–1.39(m,6H),1.13(t,J=7.2Hz,3H),0.99(s,3H). 13 CNMR (125MHz, CDCl3) δ173.0,172.7,158.8,150.9,150.3,148.7,138.1, 135.8,129.8,128.7,113.5,111.0,110.5,110.1,69.1,69.0,66.6,63.5 ,61.2,54.2,52.8,52.7,52.6,52.3,48.3,42.4,40.1,40.0,35.5,34.1,33.3,33.2,29.1,29.0,27.3,20.1,16.1,11.7.HRMS(ESI)m / z:calcdforC 42 H 61 N5O9S[M+H] + ,812.4263;found,812.4261.

[0246] Example 27: Preparation of Compound 27 (20c)

[0247]

[0248] The preparation method of 4-(3-((4-((1-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-4-oxobutanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 25, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0249] In this example, a colorless transparent liquid was obtained with a yield of 52%.

[0250] 1 HNMR(500MHz, CDCl3)δ8.06(dd,J=1.1,8.5Hz,2H),7.79–7.73(m,1H),7.63(td,J=1.6,7.7Hz,2H),5.80 (dd,J=10.8,17.5Hz,1H),5.04(s,1H),4.95–4.83(m,4H),4.78(s,1H),4.51(t,J=6.1Hz,2H),4.30(t,J =6.1Hz,2H),3.90(d,J=6.4Hz,2H),2.92(tt,J=11.2,41.2Hz,6H),2.63(s,6H),2.59–2.39(m,8H),2.22 (p,J=6.2Hz,4H),2.07(t,J=11.5Hz,1H),1.96(t,J=13.4Hz,1H),1.72(t,J=11.6Hz,1H),1.68–1.39(m,

[0251] 10H), 1.14 (t, J = 7.2Hz, 3H), 0.99 (s, 3H). 13 CNMR(125MHz,CDCl3)δ172.4,172.3,

[0252] 159.0,150.9,150.3,148.7,138.1,135.8,129.8,128.7,113.5,110.9,110.6,110.1,69.3,68.0,66.6,63.5,60.6,54.2,52.8, 52.8,52.6,52.3,48.2,42.4,40.1,40.0,35.5,34.1,32.0,29.1,29.0,28.9,28.0,27.3,16.0,11.7.HRMS(ESI)m / z:calcdforC 42 H 61 N5O9S[M+H] + ,812.4263;found,812.4269.

[0253] Example 28: Preparation of Compound 28 (20d)

[0254]

[0255] The preparation method of 4-(3-((5-((1-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-5-oxopentanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 26, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0256] In this example, a colorless transparent liquid was obtained with a yield of 44%.

[0257] 1HNMR(500MHz, CDCl3) δ8.06(dt,J=1.5,8.6Hz,2H),7.76(tt,J=1.2,7.2Hz,1H),7.63(td,J=1.7,7.6Hz,2H),5.80(dd,J= 10.8,17.5Hz,1H),5.03(s,1H),4.95–4.83(m,4H),4.77(s,1H),4.50(t,J=6.1Hz,2H),4.27(t,J=6.1Hz,2H),3.92(d,J= 6.2Hz,2H),3.03–2.87(m,3H),2.83(t,J=10.6Hz,2H),2.66–2.43(m,8H),2.38(dt,J=7.4,12.3Hz,5H),2.21(dq,J=4.9, 5.5,11.2Hz,3H),2.07(t,J=11.5Hz,1H),1.95(p,J=7.3Hz,3H),1.86–1.39(m,14H),1.11(t,J=7.2Hz,3H),0.99(s,3H). 13 CNMR(125MHz,CDCl3)δ173.0,172.9,158.9,151.0,150.3,148.9,138.1,1 35.8,129.8,128.6,113.3,110.8,110.5,110.1,69.1,68.0,66.7,63.6,60 .4,54.2,53.2,52.9,52.6,52.4,48.2,42.4,40.1,39.9,35.5,34.1,33.3 ,33.2,29.1,29.1,28.0,27.3,20.2,16.1,12.0.HRMS(ESI)m / z:calcdforC 43 H 63 N5O9S[M+H] + ,826.4419;found,826.4469.

[0258] Example 29: Preparation of Compound 29 (20e)

[0259]

[0260] The preparation method of 4-((4-((4-((1-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-4-oxobutanoyl)oxy)but-2-ynyl-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 25, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0261] In this example, a colorless transparent liquid was obtained with a yield of 31%.

[0262] 1 HNMR(500MHz, CDCl3)δ8.09–8.04(m,2H),7.79–7.74(m,1H),7.66–7.61(m,2H),5.80(dd,J=10.8,17.5Hz,1H),5 .09(t,J=1.6Hz,2H),5.02(s,1H),4.94–4.83(m,4H),4.79–4.73(m,3H),3.95(d,J=6.2Hz,2H),3.00(d,J=13.8Hz ,1H),2.92(q,J=13.4Hz,2H),2.82(t,J=10.4Hz,2H),2.70–2.36(m,15H),2.19(dd,J=3.1,12.8Hz,1H),2.07(t, J=11.5Hz,1H),1.94(t,J=11.5Hz,1H),1.75–1.57(m,6H),1.52–1.41(m,4H),1.10(t,J=7.2Hz,3H),0.98(s,3H). 13 CNMR (125MHz, CDCl3) δ172.1,171.6,158.0,151.0,150.3,148.8,138.0, 135.8,129.8,128.8,113.4,110.8,110.7,110.1,83.9,78.8,69.4,66.7 ,63.6,58.7,54.2,53.0,52.9,52.6,52.4,52.2,48.2,42.4,40.1,40.0,35.5,34.1,29.0,29.0,29.0,27.3,16.1,11.9.HRMS(ESI)m / z:calcdforC 43 H 59 N5O9S[M+H] + ,822.4106;found,822.4107.

[0263] Example 30: Preparation of Compound 30 (20f)

[0264]

[0265] The preparation method of 4-((4-((5-((1-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperidin-4-yl)methoxy)-5-oxopentanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 26, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0266] In this example, a colorless transparent liquid was obtained with a yield of 55%.

[0267] 1 HNMR (500MHz, CDCl3) δ8.07(d,J=8.1Hz,2H),7.77(t,J=7.5Hz,1H),7.63(t,J=7.6Hz,2H),5.80(dd,J= 10.8,17.4Hz,1H),5.10(s,2H),5.03(s,1H),4.94–4.83(m,4H),4.76(d,J=16.1Hz,3H),3.93(d,J=5.8H z,2H),2.93(ddt,J=11.4,38.2,51.7Hz,6H),2.64(d,J=13.9Hz,2H),2.55–2.34(m,11H),2.20(d,J=12. 3Hz,1H),2.09–2.04(m,1H),1.99–1.91(m,3H),1.75–1.33(m,13H),1.12(t,J=6.9Hz,3H),0.99(s,3H). 13 CNMR (125MHz, CDCl3) δ172.9,172.2,158.0,151.0,150.3,148.8,138.0,13 5.8,129.8,128.8,113.4,110.8,110.7,110.1,84.1,78.7,69.1,66.7,63.6 ,58.7,54.2,53.1,52.9,52.6,52.4,52.0,48.3,42.4,40.1,40.0,35.5,34. 1,33.2,33.1,29.1,29.1,27.3,20.1,16.1,11.9.HRMS(ESI)m / z:calcdforC 44 H 61 N5O9S[M+H]+ ,836.4263;found,836.4268.

[0268] Example 31: Preparation of Compound 31 (21a)

[0269]

[0270] The preparation method of 4-(2-((4-(2-(4-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-4-oxobutanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 1, except that N-methyl-2-hydroxyethylamine in (III) is replaced by 1-(2-hydroxyethyl)piperazine.

[0271] In this example, a colorless transparent liquid was obtained with a yield of 52%.

[0272] 1 HNMR(500MHz, CDCl3)δ8.05(dd,J=1.2,8.5Hz,2H),7.77–7.72(m,1H),7.61(td,J=1.7,7.6Hz,2H),5.78( dd,J=10.8,17.5Hz,1H),5.02(s,1H),4.93–4.81(m,4H),4.76(s,1H),4.63–4.60(m,2H),4.50(dd,J=3.7 ,5.4Hz,2H),4.20(t,J=6.0Hz,2H),3.00(d,J=13.8Hz,1H),2.95–2.87(m,2H),2.72–2.59(m,9H),2.57–2 .36(m,14H),2.35–2.30(m,1H),2.24–2.01(m,3H),1.62–1.38(m,6H),1.10(t,J=7.2Hz,3H),0.97(s,3H). 13 CNMR (125MHz, CDCl3) δ172.2,172.1,158.8,150.9,150.2,148.4,138.1,135.8,129.8,128.7,113.6,110.9,110.5,110.2,68.9,66 .3,63.5,62.4,61.5,56.7,53.6,53.1,52.9,52.4,48.3,42.4,40.1,40.0,34.0,29.1,29.0,27.3,16.0,11.8.HRMS(ESI)m / z:calcd forC41 H 60 N6O9S[M+H] + ,813.4215;found,813.4222.

[0273] Example 32: Preparation of Compound 32 (21b)

[0274]

[0275] The preparation method of 4-(2-((5-(2-(4-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-5-oxopentanoyl)oxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 31, except that succinic anhydride in (V) is replaced by glutaric anhydride.

[0276] In this example, a colorless transparent liquid was obtained with a yield of 38%.

[0277] 1 HNMR (500MHz, CDCl3) δ8.08–8.04(m,2H),7.78–7.74(m,1H),7.62(td,J=1.7,7.6Hz,2H),5.79(dd,J=10.8,17.5Hz, 1H),5.03(s,1H),4.94–4.82(m,4H),4.77(s,1H),4.64–4.61(m,2H),4.50(dd,J=3.7,5.4Hz,2H),4.20(t,J=6.0Hz,2 H),3.01(d,J=13.8Hz,1H),2.92(q,J=13.4Hz,2H),2.67–2.58(m,4H),2.55–2.27(m,20H),2.18(dd,J=3.1,12.8Hz, 1H),2.10–2.03(m,2H),2.00–1.95(m,2H),1.64–1.56(m,2H),1.52–1.38(m,4H),1.10(t,J=7.2Hz,3H),0.98(s,3H). 13CNMR (125MHz, CDCl3) δ172.9,172.7,158.8,150.9,150.2,148.4,138.1,135.8,129.8,128.7,113.6,110.9,110.5,110.2,69.0,66.3,63.5 ,61.9,61.2,56.8,53.6,53.1,52.9,52.8,52.4,48.3,42.4,40.1,40.0,34.0,33.2,33.1,27.3,20.1,16.0,11.8.HRMS(ESI)m / z:calcdforC 42 H 62 N6O9S[M+H] + ,827.4372;found,827.4323.

[0278] Example 33: Preparation of Compound 33 (21c)

[0279]

[0280] The preparation method of 4-(3-((4-(2-(4-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-4-oxobutanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 31, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0281] In this example, a colorless transparent liquid was obtained with a yield of 70%.

[0282] 1HNMR (500MHz, CDCl3) δ8.08–8.04(m,2H),7.78–7.73(m,1H),7.63(t,J=7.9Hz,2H),5.80(dd,J=10.8,17.5H z,1H),5.04(s,1H),4.95–4.83(m,4H),4.78(s,1H),4.51(t,J=6.1Hz,2H),4.30(t,J=6.1Hz,2H),4.18(t,J =6.0Hz,2H),3.02(d,J=13.8Hz,1H),2.98–2.89(m,2H),2.71–2.28(m,24H),2.21(qd,J=3.6,11.3Hz,4H),2 .08(d,J=11.6Hz,1H),1.60(dd,J=8.0,12.1Hz,2H),1.53–1.38(m,4H),1.15(t,J=7.2Hz,3H),0.99(s,3H). 13 CNMR (125MHz, CDCl3) δ172.3,172.3,158.9,150.9,150.2,148.4,138.1,135.7,129.8,128.7,113.6,110.8,110.6,110.1,68.0,66.3,63.5 ,62.3,60.6,56.7,53.6,53.1,53.0,52.9,52.4,48.3,42.4,40.1,39.9,34.0,29.1,29.1,28.0,27.2,16.0,11.8.HRMS(ESI)m / z:calcdforC 42 H 62 N6O9S[M+H] + ,827.4372;found,827.4326.

[0283] Example 34: Preparation of Compound 34 (21d)

[0284]

[0285] The preparation method of 4-(3-((5-(2-(4-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-yl))-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-5-oxopentanoyl)oxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 32, except that ethylene glycol in (IV) is replaced by propylene glycol.

[0286] In this example, a colorless transparent liquid was obtained with a yield of 58%.

[0287] 1 HNMR (500MHz, CDCl3) δ8.05–8.03(m,2H),7.77–7.72(m,1H),7.62(td,J=1.7,7.6Hz,2H),5.78(dd,J=10.8,17 .5Hz,1H),5.02(s,1H),4.93–4.82(m,4H),4.77(s,1H),4.49(t,J=6.1Hz,2H),4.26(t,J=6.1Hz,2H),4.18(t,J =6.0Hz,2H),3.01(d,J=13.8Hz,1H),2.96–2.88(m,2H),2.67–2.57(m,5H),2.57–2.34(m,19H),2.26–2.17(m, 4H),2.03(s,1H),1.95–1.91(m,2H),1.64–1.56(m,2H),1.51–1.39(m,4H),1.12(t,J=7.2Hz,3H),0.97(s,3H). 13 CNMR(125MHz, CDCl3)δ172.9,158.9,150.8,150.2,148.4,138.1,135.8,129.8,128.6,113.7,111.0,110.5,110.2,68.1,66.3,63.4,61.9, 60.4,56.8,53.6,53.1,52.7,52.6,52.3,48.3,42.4,40.0,40.0,34.0,33.3,33.2,28.0,27.3,20.1,16.0,11.6.HRMS(ESI)m / z:calcdforC 43 H 64 N6O9S[M+H] + ,841.4528;found,841.4536.

[0288] Example 35: Preparation of Compound 35 (21e)

[0289]

[0290] The preparation method of 4-((4-((4-(2-(4-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-)yl)-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-4-oxobutanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 31, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0291] In this example, a colorless transparent liquid was obtained with a yield of 37%.

[0292] 1 HNMR(500MHz, CDCl3)δ8.06(dd,J=1.1,8.4Hz,2H),7.78–7.73(m,1H),7.66–7.59(m,2H),5.79(dd,J=10. 8,17.5Hz,1H),5.09(t,J=1.6Hz,2H),5.02(s,1H),4.94–4.81(m,4H),4.78–4.73(m,3H),4.21(t,J=6.0H z,2H),3.01(d,J=13.8Hz,1H),2.91(q,J=13.4Hz,2H),2.69–2.59(m,8H),2.56–2.24(m,17H),2.21–2.15 (m,1H),2.06(t,J=12.1Hz,1H),1.63–1.55(m,2H),1.51–1.38(m,4H),1.08(t,J=7.2Hz,3H),0.98(s,3H). 13 CNMR (125MHz, CDCl3) δ172.1,171.6,158.0,151.0,150.3,148.5,138.0,135.8,129.8,128.8,113.6,110.9,110.7,110.2,83.9,78.8,66.3,63 .6,62.4,58.7,56.7,53.6,53.2,53.0,52.9,52.4,52.2,48.3,42.4,40.1,40.0,34.1,29.0,28.9,27.3,16.1,11.9.HRMS(ESI)m / z:calcdforC 43 H 60 N6O9S[M+H] + ,837.4215;found,837.4232.

[0293] Example 36: Preparation of Compound 36 (21f)

[0294]

[0295] The preparation method of 4-((4-((5-(2-(4-(2-((1R,2S,5R)-5-(3-(4-ethylpiperazin-1-yl)prop-1-en-2-)yl)-2-methyl-2-vinylcyclohexyl)allyl)piperazin-1-yl)ethoxy)-5-oxopentanoyl)oxy)but-2-yn-1-yl)oxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide is basically the same as that in Example 32, except that the ethylene glycol in (IV) is replaced by 1,4-butynediol.

[0296] In this example, a colorless transparent liquid was obtained with a yield of 52%.

[0297] 1 HNMR (500MHz, CDCl3) δ8.09–8.03(m,2H),7.79–7.72(m,1H),7.66–7.58(m,2H),5.78(dd,J=10.8,17.5Hz,1H),5.09(t ,J=1.7Hz,2H),5.02(s,1H),4.93–4.82(m,4H),4.77(d,J=1.9Hz,1H),4.73(t,J=1.6Hz,2H),4.19(t,J=5.9Hz,2H),3. 01(d,J=13.8Hz,1H),2.95–2.87(m,2H),2.66–2.58(m,4H),2.57–2.26(m,21H),2.17(dd,J=3.1,12.8Hz,1H),2.05(t, J=10.5Hz,1H),1.98–1.92(m,2H),1.59(dd,J=7.1,12.5Hz,2H),1.51–1.39(m,4H),1.10(t,J=7.2Hz,3H),0.97(s,3H). 13 CNMR (125MHz, CDCl3) δ172.8,172.2,158.0,150.9,150.2,148.4,137.9, 135.8,129.8,128.7,113.6,110.9,110.7,110.2,84.1,78.6,66.3,63.5 ,61.9,58.7,56.8,53.6,53.1,52.9,52.8,52.4,51.9,48.3,42.4,40.1,40.0,34.0,33.2,33.0,27.3,20.0,16.0,11.8.HRMS(ESI)m / z:calcdfor C 44 H 62N6O9S[M+H] + ,851.4372;found,851.4381.

[0298] Pharmacodynamic experiments

[0299] 1. In vitro nitric oxide release test

[0300] 1.1 Experimental equipment and reagents

[0301] instrument

[0302] Multifunctional microplate reader (MD Spectramac M3, USA)

[0303] Clean bench (SW-CJ-1FD, Suzhou, China)

[0304] Reagents

[0305] Carbon monoxide (NO) assay kit (A013-2-1, Nanjing Jiancheng Bioengineering Institute, China)

[0306] 1.2 Experimental methods

[0307] 1. Sample pretreatment: Dilute the sample to 100 μM, mix thoroughly, and take 160 μL for operation.

[0308] 2 Operation table:

[0309]

[0310] Calculation method:

[0311] The calculation formula of NO content in tissue samples is:

[0312]

[0313] 1.3 Test results

[0314] Table 1. In vitro NO release results of β-elemene nitric oxide donor derivatives obtained in Examples 1-36

[0315]

[0316]

[0317] As can be seen from the data in Table 1, with the exception of Examples 12 and 18, which exhibited higher NO release, the remaining compounds exhibited lower NO release. NO release for both compounds was time-dependent, with a significant increase after 30 minutes. Furthermore, the NO content of 13-pyrazole-β-elemene compounds was higher than that of 13-N-ethylpiperazine-β-elemene compounds. The higher NO release levels of these two compounds were also reflected in their subsequent stronger antiproliferative activity.

[0318] 2. In vitro anti-tumor activity evaluation test

[0319] 2.1 Experimental equipment

[0320] Clean bench (Suzhou Purification SW-CJ-1FD);

[0321] CO2 incubator (SANYOXD-101);

[0322] Biological inverted microscope (OLYMPUSIX51);

[0323] Microplate reader (BioTek Instruments EL-x800);

[0324] 2.2 Experimental methods

[0325] 1) Digest and count the cells and prepare cell suspension (K562, CCRF-CEM 5×10 5 / mL, and the remaining 3.5×10 4 / mL), 100 μL of cell suspension was added to each well of a 96-well cell culture plate;

[0326] 2) The 96-well cell culture plate was placed in a 37°C, 5% CO2 incubator for 24 h;

[0327] 3) Dilute the drug to the desired working concentration using culture medium, add 100 μL of the corresponding drug-containing culture medium to each well, and set up a negative control group at the same time;

[0328] 4) Place the 96-well cell culture plate in a 37°C, 5% CO2 incubator for 72 h;

[0329] 5) Stain the 96-well plate with CCK-8 at λ = 450 nm and measure the OD value;

[0330] 1) Add 10 μL of CCK-8 to each well and continue incubation in the incubator for 1-2 hours;

[0331] 2) Gently mix on a shaker for 10 minutes to remove bubbles in the 96-well plate;

[0332] 3) λ = 450 nm, read the OD value of each well with a microplate reader, and calculate the inhibition rate.

[0333] 6) Calculate the inhibition rate of each group.

[0334]

[0335] 2.3 Experimental Results

[0336] Table 2. Inhibitory effect of the compounds of Examples 1-36 on the antiproliferative activity of four human cancer cell lines (1 and 10 μM)

[0337]

[0338]

[0339] As shown in Table 2, all compounds exhibited stronger antiproliferative activity than β-elemene, indicating that the introduction of an NO donor further enhances its antiproliferative activity. Furthermore, at concentrations of 1 and 10 μM, most compounds exhibited significantly stronger inhibitory effects against K562 and CCRF-CEM cells than against A375 and SK-MEL-2 cells. This may be due to the presence of metabolites of the NO donor derivatives of β-elemene in the blood, which possess antitumor properties. Consequently, these compounds exhibited greater sensitivity against K562 and CCRF-CEM cell lines than against A375 and SK-MEL-2 cell lines.

[0340] Table 3. IC values ​​of the compounds of Examples 1-18 for antiproliferative activity against two human cancer cell lines 50 (μM)

[0341]

[0342] The results are shown in Table 3. All compounds had stronger antiproliferative activities than β-elemene, and the IC 50 The values ​​were even less than 1 μM. Among them, Examples 12 (17f) and 18 (18f) showed inhibitory activities against K562 and CCRF-CEM cells that were 100 times and 90 times greater than those of β-elemene, respectively. The more active Examples 12 (17f) and 18 (18f) also exhibited higher NO release levels, demonstrating a synergistic antitumor effect. These results suggest that the introduction of NO donors may enhance the antitumor activity of β-elemene.

[0343] 3. In vivo antitumor activity against human leukemia K562 cell xenografts in nude mice

[0344] 3.1 Test animals and experimental equipment

[0345] Source, species, and strain: BALB / c nude mice, provided by Shanghai Lingchang Biotechnology Co., Ltd.

[0346] Age: 6-8 weeks

[0347] Gender: Female

[0348] Number of animals: 6 animals per group, 3 groups in total, 18 animals in total

[0349] Experimental instruments:

[0350] Electronic scale (T1000, manufactured by Changshu Shuangjie Testing Instrument Factory);

[0351] Analytical balance (Mettler Toledo ME204);

[0352] 3.2 Grouping and Dosage Regimen

[0353] Model group: Inject the same amount of solvent into the tail vein, 100 μL / 10g, once a day, and observe for 28 days

[0354] β-elemene group: Tail vein injection, 30 mg / kg, 100 μL / 10 g, once a day, continuous observation for 28 days

[0355] Example 18 (18f) group: tail vein injection, 30 mg / kg, 100 μL / 10 g, once a day, continuous observation for 28 days

[0356] 3.3 Model preparation

[0357] The cultured human leukemia K562 cell suspension was collected at a concentration of 5×10 7 Each mouse was inoculated with 0.1 mL of the drug subcutaneously in the right axilla.

[0358] 3.4 Grouping and Dosing

[0359] The diameter of the transplanted tumor in mice was measured with a vernier caliper. The tumor grew to 50-100 mm. 3 At the same time, the animals were randomly divided into groups. At the same time, each group of mice began to receive medication according to the dosing schedule described in Groups and Dosing Schedule. After the experiment, the mice were immediately sacrificed, and the tumor masses were surgically removed and weighed.

[0360] 3.5 Experimental Results

[0361] Within 4 weeks of administration, the body weight of the model group, β-elemene group and Example 18 (18f) group gradually increased with the increase in treatment time, indicating that β-elemene and Example 18 (18f) groups had no obvious toxicity. However, after weighing the tumors at the end of the experiment, it was found that the growth of tumor volume in the model group was seriously exaggerated with the increase in treatment time. In contrast, the growth of tumor volume in the β-elemene group and Example 18 (18f) group was not significantly inhibited. In addition, the tumor weight of the β-elemene group and Example 18 (18f) group was significantly lower than that of the model group, indicating that they both effectively inhibited tumor growth. As expected, the anti-tumor effect of the Example 18 (18f) group was better than that of the β-elemene group. In terms of inhibiting tumor growth, although both can inhibit tumor growth, the inhibitory effect of Example 18 (18f) is about 20% stronger than that of β-elemene.

[0362] The above results indicate that the introduction of nitrogen-containing groups and NO donors effectively enhances the anti-tumor effect of β-elemene in vivo, which is a feasible strategy for the development of anti-tumor drugs based on β-elemene.

[0363] In addition, it should be understood that after reading the above description of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A 13-heterocyclic-β-elemene nitric oxide donor derivative, characterized in that: The structure of the 13-heterocyclic-β-elemene nitric oxide donor derivative is shown in formula (I): in: R 1 for R 2 Selected from The oxygen atom is attached to the carbonyl group; R 3 、R 4 They are independently selected from -CH2CH2-, -CH2CH2CH2-, -CH2C≡C-, -C≡CCH2-, -CH2CH2CH2CH2-, -CH2C≡CCH2-, -CH2CH2CH2CH2CH2-, -CH2C≡CCH2CH2-, -CH2CH2C≡CCH2-, -CH2CH2C≡CCH2-, -CH2CH2C≡CCH2CH2-, and are connected to the two end structures from left to right.

2. A 13-heterocyclic-β-elemene nitric oxide donor derivative according to claim 1, characterized in that: The 13-heterocyclic-β-elemene nitric oxide donor derivative is selected from compounds 1 to 36 shown in the following structures:

3. Use of the 13-heterocyclic-β-elemene nitric oxide donor derivative according to claim 1 or 2 in the preparation of anti-tumor drugs, wherein the tumors include leukemia and melanoma.

4. An anti-tumor drug comprising a safe and effective amount of the 13-heterocyclic-β-elemene nitric oxide donor derivative according to claim 1 or 2; the tumors include leukemia and melanoma.

5. The antitumor drug according to claim 4, characterized in that It also includes pharmaceutically acceptable salts and excipients or carriers.

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