CN111320617A - Phenyloxadiazole derivatives and preparation method and application thereof - Google Patents

Abstract

The invention discloses a phenyl oxadiazole derivative and a preparation method and application thereof, belonging to the field of medicinal chemistry. The phenyl oxadiazoleThe structural general formula of the derivative is shown as formula I or formula II:

in the formula I, A is N or CH; m is 0, 1 or 2; z is (CH)₂)_nN is 2,3 or 4; r₁Is hydrogen, unsubstituted C_1‑5Alkyl or substituted C_1‑5An alkyl group; r₂And R₃Each independently selected from fluorine, chlorine, bromine and iodine; in the formula I, Z is (CH)₂)_n，R₂And R₃Each independently selected from fluorine, chlorine, bromine and iodine. The derivative is applied to preparing the medicines for preventing or treating pain diseases, and has the beneficial effects of small side effect, high response rate and difficulty in generating drug resistance and addiction.

Description

Phenyloxadiazole derivatives and preparation method and application thereof

technical field

The invention belongs to the field of medicinal chemistry, and in particular relates to a phenyloxadiazole derivative and a preparation method and application thereof.

Background technique

Pain is an unpleasant sensory and emotional experience that accompanies existing or potential tissue damage and is the body's protective response to environmental stimuli, but prolonged or excessive pain can trigger a range of organisms such as shock Functional changes seriously affect people's normal physiological life, especially chronic pain can often make patients feel unbearable, and is the cause of disease, death and disability. Neuralgia is one of the pain-related diseases that seriously harms the lives of patients. About 8% of the world's people are affected by this disease. The onset of neuralgia is sudden, severe, and difficult to cure. Although researchers have done a lot of related work in the field of basic research and clinical research, there are still huge challenges in clinical research on neuralgia and analgesic mechanisms. Due to the complex causes and pathogenesis of neuralgia, there are no broad-spectrum and specific targeted drugs for neuralgia. At present, most of the drugs used in the clinical treatment of neuralgia are some drugs for the treatment of depression, epilepsy and other diseases that have been found to have neuralgia analgesic activity in clinical applications. Most of these drugs are used off-label for the treatment of neuralgia, the analgesic effect is often lower than expected, and the response rate to treatment varies significantly among individuals.

At present, there are no specific methods or effective drugs in clinical treatment of neuralgia. Neuropsychiatric drugs are generally used for adjuvant treatment in clinical practice. The main drugs used are: 1) Neurotransmitter reuptake inhibitors such as antidepressant drugs (such as Triline and duloxetine), these drugs have more side effects and the response rate is not high; 2) calcium channel α2 _- δ ligand antiepileptic drugs (such as gabapentin and pregabalin), such drugs have limited effectiveness , about 40%, and individual differences are obvious; 3) opioid receptor analgesics; such drugs are prone to drug resistance and addiction, and the adverse reactions are obvious. Other new targets, such as NMDA receptor antagonists, α2 _- adrenergic receptor agonists, antiarrhythmic drugs, etc., have also developed new drug research in recent years, but most of them are still in the early stage of research, and a few have entered the first and second Phase clinical. Therefore, it is of great significance to search for new neuralgia analgesic drugs based on new action mechanisms and targets, and have good therapeutic effects on neuralgia.

The sigma-1 receptor (σ ₁ receptor) is an emerging drug target in recent years, which exhibits excellent biological activity especially in neuralgia analgesia. σ ₁ receptors are mainly distributed in the central nervous system, but also widely distributed in peripheral organs. Research on the regulation of sigma ₁ receptors in human pain began in the 1990s, when it was found that sigma ₁ receptor antagonists can sensitize the analgesic activity of opioid analgesics, while sigma ₁ agonists attenuate this phenomenon. In acute pain experiments, antisense oligodeoxynucleotides of σ ₁ receptors can significantly enhance the analgesic effects of morphine and opioid μ receptor agonists. After that, more experiments proved that the σ ₁ receptor works by directly interacting with the opioid μ receptor. On the other hand, σ ₁ receptor antagonists themselves have analgesic effects on pain, especially neuralgia. In the formalin-induced pain experiment using σ ₁ receptor knockout mice, there was no obvious pain response in both phases I and II. The same phenomenon occurs in a model of neuropathic hyperalgesia induced by capsaicin stimulation. σ ₁ receptor knockout mice also showed insensitive behavior in paclitaxel-induced cold-induced hyperalgesia and mechanical stimulation hyperalgesia models and sciatic nerve ligation neuralgia models. When the normal mice were subjected to pain model experiments with antagonists of σ ₁ receptors, they showed a phenomenon of desensitization similar to that of σ ₁ receptor knockout mice. Experiments have shown that haloperidol and its metabolites I and II, which have σ ₁ receptor antagonistic effects, have obvious analgesic effects in various neuralgia models. The σ ₁ receptor antagonists BD-1063 and NE-100 also had similar results. At present, S1RA is at the forefront of research on sigma-1 receptor antagonists, which has high affinity (Ki=17nM) for σ ₁ receptors and high selectivity. At present, S1RA is used in experiments for the treatment of various pains and has entered the clinical stage, among which the experiment for the treatment of neuralgia alone has entered the clinical phase II.

Therefore, the search for selective σ ₁ receptor antagonists for anti-pain therapy has important scientific value and social significance for clinical treatment of pain and neuralgia.

SUMMARY OF THE INVENTION

The present invention solves the technical problems of large side effects, low response rate and easy generation of drug resistance and addiction in the prior art drugs for treating pain.

According to the first aspect of the present invention, there is provided a phenyloxadiazole derivative, and the general structural formula of the phenyloxadiazole derivative is as shown in formula I or formula II:

Wherein, in the formula I, A is N or CH; m is 0, 1 or 2; Z is (CH ₂ ) _n , and the n is 2, 3 or 4; R ₁ is hydrogen, unsubstituted C _{1 -5} alkyl or substituted C _1-5 alkyl; R ₂ and R ₃ are each independently selected from fluorine, chlorine, bromine and iodine;

Wherein, in the formula I, Z is (CH ₂ ) _n , and R ₂ and R ₃ are each independently selected from fluorine, chlorine, bromine and iodine.

Preferably, the substituted C _1-5 alkyl is a fluorine, chlorine, bromine or iodine substituted C _1-5 alkyl.

According to another aspect of the present invention, the preparation method of described phenyloxadiazole derivatives is provided, and the reaction formula of described preparation method is as follows:

The reaction formula of the preparation method is as follows:

wherein, A is N or CH; m is 0, 1 or 2; Z is (CH ₂ ) _n , and the n is 2, 3 or 4; R ₁ is hydrogen, unsubstituted C _1-5 alkyl or substituted C _1-5 alkyl; R ₂ and R ₃ are each independently selected from fluorine, chlorine, bromine and iodine.

According to another aspect of the present invention, there is provided a phenyloxadiazole derivative salt, the salt is an anion salt formed by the phenyloxadiazole derivative; the phenyloxadiazole derivative Derivative salts are hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, tartrate , maleate, fumarate, mesylate, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate or p-toluenesulfonate.

According to another aspect of the present invention, there is provided a pharmaceutical composition, the pharmaceutical composition contains the phenyloxadiazole derivatives according to claim 1 or 2, or the pharmaceutical composition contains the phenyloxadiazole derivative according to claim 3 The phenyloxadiazole derivatives described above.

Preferably, the pharmaceutical composition further contains adjuvants or carriers;

Preferably, the adjuvant is a binder, an excipient, a disintegrant, a lubricant or a sweetener;

Preferably, the carrier is grease.

According to another aspect of the present invention, there is provided the use of the phenyloxadiazole derivatives for the preparation of medicines for preventing or treating pain diseases; or the phenyloxadiazole derivative salts used for The application of preparing a medicine for preventing or treating pain diseases.

Preferably, the pain-like disease is neuralgia, cancer pain, angina pectoris, thromboangiitis pain or chest and abdominal pain.

Preferably, the dosage form of the medicine is tablet, lozenge, capsule, suspension, syrup, ointment, lotion or injection.

Overall, compared with the prior art, the above technical solutions conceived by the present invention mainly have the following technical advantages:

(1) The application of the derivatives in the present invention for the preparation of medicines for preventing or treating pain-like diseases has the beneficial effects of less side effects, high response rate, and difficulty in producing drug resistance and addiction.

(2) The derivatives and their salts of the present invention have high affinity to σ ₁ receptors, but low affinity to σ ₂ receptors, and have selective antagonism to σ ₁ receptors, indicating that they have analgesic activity. In addition, animal test results also show that these compounds can significantly improve both formalin-induced phase I and phase II pain. Since these in vitro action targets and in vivo pharmacological models are closely related to the sigma ₁ receptor-mediated nervous system regulation response, especially pain, the compounds involved in the present invention have the effect of treating pain, especially neuralgia.

Detailed ways

In order to make the objectives, technical solutions and advantages of the present invention more clear, the following examples are used to further describe the present invention in detail. It should be understood that the specific embodiments described herein are only used to explain the present invention, but not to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

The present invention provides a pharmaceutical composition comprising a compound of formula I or II or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable auxiliary materials (such as carrier and/or excipient, etc.), the pharmaceutical composition is Contains compounds sufficient to produce pain, neuralgia analgesic activity.

An effective dose of a compound of this invention can be administered orally with, for example, an inert diluent or some kind of carrier. It can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapy, the compounds of this invention can be used with excipients and in the form of tablets, troches, capsules, suspensions, syrups and the like. These formulations should contain at least 0.5% by weight of the active compound of the present invention, but may vary depending on the particular dosage form, from 4% to about 70% by weight conveniently. The amount of active compound in such compositions should result in an appropriate dosage. An oral unit dose of the preferred compositions and formulations of the present invention contains 1.0 to 300 mg of the active compound of the present invention.

The compounds provided by the present invention and their pharmaceutically acceptable salts, solvates and hydrates can be combined with pharmaceutically acceptable carriers or diluents to form pharmaceutical preparations. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile aqueous or organic solutions.

The amount of the compound of the present invention to be used will depend on the type and severity of the disease or disorder, and on the characteristics of the subject, such as general health, age, sex, weight and drug tolerance. The skilled artisan will be able to determine appropriate dosages based on these and other factors. Effective doses of commonly used CNS drugs are well known to the skilled artisan. The total daily dose is usually between about 0.05 mg and 2000 mg.

The present invention relates to pharmaceutical compositions which provide about 0.01 to 1000 mg of active ingredient per unit dose. The compositions can be administered by any suitable route, such as orally in capsule form, parenterally in the form of injection solutions, topically in the form of ointments or lotions, rectally in the form of suppositories, in the form of patch delivery systems Transdermal administration.

The compounds provided herein can be combined with suitable solid or liquid carriers or diluents to form capsules, tablets, pills, powders, syrups, solutions and the like. Tablets, pills, capsules, etc. contain from about 0.01 to about 99 percent by weight of the active ingredient and binders such as gelatin, cornstarch, acacia; excipients such as calcium hydrogen phosphate; disintegrants such as cornstarch, potato starch or algae acids; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose. When the preparation is in the form of a capsule, it may contain, in addition to the above-mentioned types of materials, a liquid carrier such as a fat and oil.

For parenteral administration, the compounds provided herein can be combined with sterile aqueous or organic vehicles to form injectable solutions or suspensions.

Compounds of general formula I or formula II may contain chiral centers and thus may exist in different enantiomeric and diastereomeric forms. The present invention relates to all optical isomers and to all stereoisomers of compounds of general formula (I), as racemic mixtures and to individual enantiomeric and diastereomeric forms of such compounds, and to such compounds as All pharmaceutical compositions and methods of treatment as defined above containing or using them.

In addition, the compounds provided by the present invention and the pharmaceutical compositions consisting of the compounds can be used for the treatment and prevention of pain; the pain refers to acute pain, such as soft tissue and joint acute injury pain, postoperative pain, obstetric pain, acute herpes zoster pain , gout, etc.; chronic pain such as soft tissue and joint strain or degenerative pain, discogenic pain, neurogenic pain, etc.; intractable pain such as trigeminal neuralgia, post-herpetic neuralgia, intervertebral disc herniation, intractable headache, etc. ; cancer pain such as advanced tumor pain, tumor metastasis pain, etc.; special pain categories such as thromboangiitis, refractory angina pectoris, idiopathic thoracic and abdominal pain, etc.

The in vitro receptor binding test shows that the compounds involved in the present invention have high affinity to σ ₁ receptors, but low affinity to σ ₂ receptors. Selective antagonism of σ ₁ receptors, indicating potential for analgesic activity.

In addition, animal test results also show that these compounds can significantly improve both formalin-induced phase I and phase II pain. Since these in vitro action targets and in vivo pharmacological models are closely related to the sigma ₁ receptor-mediated nervous system regulation response, especially pain, the compounds involved in the present invention have the potential to treat pain, especially neuralgia.

First Aspect: Examples of Synthetic Aspects

Example 1

The preparation method of 3-(3,4-difluorophenyl)-5-(4-(4-methylpiperidin-1-yl)butyl)-1,2,4-oxadiazole (1), Contains the following steps:

1) Take 80g of 3,4-difluorobenzonitrile in a 2000mL four-neck flask, add 500mL of ethanol and stir to dissolve completely, dissolve 81.2g of hydroxylamine hydrochloride in water, slowly add it dropwise to the reaction solution, and add 61g in batches The potassium carbonate powder was heated to 70 ° C and stirred for reaction. TLC detected the reaction completion. Most of the solvent was evaporated, water was added, and a solid was precipitated. Direct suction filtration was performed to obtain an off-white solid, and 70 mL of ethyl acetate was added to stir and dissolve at room temperature. Then 700 mL of petroleum ether was added, and a large amount of solid was precipitated, and the product was directly obtained by suction filtration, 78 g of off-white solid, and the yield was 78.2%.

2) get the first step product 30g in the single-necked bottle of 1000mL, add the acetone of 400mL and stir and dissolve under ice-bath conditions, then slowly drip the 5-chlorovaleryl chloride of 26.4g and stir the reaction, and TLC detects that the reaction is completed. Most of the solvent was directly evaporated, saturated aqueous sodium bicarbonate solution was added, and extracted with dichloromethane. The organic phase was dried and concentrated to obtain an off-white solid. After adding 20 mL of ethyl acetate to dissolve, 200 mL of petroleum ether was added. After stirring, a large amount of solid was precipitated, and 40 g of off-white solid was obtained by direct suction filtration, and the yield was 81.1%.

3) Take 30g of the second step product in a 500mL four-neck flask, add 300mL of toluene, install a water separator, heat up to 129°C, stir for 8h, and complete the TLC detection reaction. Most of the solvent was directly evaporated, the samples were mixed, rinsed with petroleum ether, and 26 g of colorless oil was obtained by column chromatography with a yield of 95.6%.

4) Take 2 g of the third step product, 0.8 g of 4-methylpiperidine, and 1.7 g of potassium carbonate, add 40 ml of acetonitrile, heat up to 80° C. for 6 hours, cool to room temperature, directly suction filtration, take the mother liquor, concentrate and mix samples, and use Elution with ethyl acetate, and column chromatography gave 1.0 g of a colorless oil with a yield of 40.7%.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.94-7.88(m, 1H), 7.85(dd, J=9.6, 4.0Hz, 1H), 7.34-7.24(m, 1H), 3.57(d, J＝11.6Hz, 2H), 3.02(dt, J＝11.5, 6.1Hz, 4H), 2.64(dd, J＝21.0, 9.9Hz, 2H), 2.22–2.08 (m, 2H), 2.08 –1.93(m,4H),1.83(d,J=14.4Hz,2H),1.66–1.52(m,1H), 1.06(d,J=6.4Hz,3H).MS(ESI)m/z 336.2( [M+H] ⁺ ).

The reaction formula is as follows:

Example 2: 3-(3,4-Difluorophenyl)-5-(2-(piperidin-1-yl)ethyl)-1,2,4-oxadiazole (2)

Substitute 5-chlorovaleryl chloride for 3-chloropropionyl chloride and 4-methylpiperidine for piperidine, and prepare the target compound according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.90 (d, J=8.8 Hz, 1H), 7.85 (dd, J=7.1, 3.8 Hz, 1H), 7.35-7.25 (m, 1H), 3.82(t, J＝7.7Hz, 2H), 3.64(s, 2H), 3.56(t, J＝7.7Hz, 2H), 2.76(s, 2H), 2.35(d, J＝11.8Hz, 2H),1.96(s,3H),1.49(s,2H).MS(ESI)m/z294.1([M+H] ⁺ ).

Example 3: 3-(3,4-Difluorophenyl)-5-(3-(piperidin-1-yl)propyl)-1,2,4-oxadiazole (3)

Substitute 5-chlorovaleryl chloride for 4-chlorobutyryl chloride and 4-methylpiperidine for piperidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.90 (d, J=9.4 Hz, 1H), 7.87-7.80 (m, 1H), 7.34-7.25 (m, 1H), 3.62 ( d, J＝11.5Hz, 2H), 3.14(t, J＝6.9Hz, 4H), 2.70(dd, J＝20.0, 10.0Hz, 2H), 2.63–2.52(m, 2H), 2.34(dd, J =26.1,12.8Hz,2H),2.00–1.73(m,2H),1.45(dd,J=25.4,12.6 Hz,2H).MS(ESI)m/z 308.2([M+H] ⁺ ).

Example 4: 4-(2-(3-(3,4-Difluorophenyl)-1,2,4-oxadiazol-5-yl)ethyl)morpholine (4)

Substitute 5-chlorovaleryl chloride for 3-chloropropionyl chloride and 4-methylpiperidine for morpholine, and prepare the target compound according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.89 (t, J=7.4Hz, 1H), 7.85 (dd, J=9.2, 3.2Hz, 1H), 7.36-7.23 (m, 1H), 4.32(d, J＝11.5Hz, 2H), 4.06(d, J＝11.9Hz, 2H), 3.80(d, J＝7.0Hz, 2H), 3.62(s, 2H), 3.55(d, J = 11.1Hz, 2H), 3.02(s, 2H). MS(ESI) m/z 296.1([M+H] ⁺ ).

Example 5: 4-(3-(3-(3,4-Difluorophenyl)-1,2,4-oxadiazol-5-yl)propyl)morpholine (5)

Substitute 5-chlorovaleryl chloride for 4-chlorobutyryl chloride and 4-methylpiperidine for morpholine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.90(d, J=9.3 Hz, 1H), 7.88-7.81(m, 1H), 7.35-7.26(m, 1H), 4.33( MS (ESI)m/z 310.1([M+H] ⁺ ).

Example 6: 3-(3,4-Difluorophenyl)-5-(2-(4-methylpiperazin-1-yl)ethyl)-1,2,4-oxadiazole (6)

Substitute 5-chlorovaleryl chloride with 3-chloropropionyl chloride and 4-methylpiperidine with 4-methylpiperazine, and prepare the target compound according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.89 (t, J=7.4Hz, 1H), 7.85 (dd, J=9.2, 3.2Hz, 1H), 7.36-7.23 (m, 1H), 4.22(d, J＝11.5Hz, 2H), 4.03(d, J＝11.9Hz, 2H), 2.07(p, J＝7.3Hz, 2H), 1.93(t, J＝11.5Hz, 2H) ,1.60(d,J＝12.8Hz,2H),1.21(td,J＝12.2,3.3Hz,2H),0.90(d,J＝6.4Hz,3H).MS(ESI)m/z 309.2([M +H] ⁺ ).

Example 7: 3-(3,4-Difluorophenyl)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (7)

The target compound was prepared according to the method of Example 1 by replacing 5-chlorovaleryl chloride with 4-chlorobutyryl chloride.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.90 (d, J=9.3 Hz, 1H), 7.88-7.81 (m, 1H), 7.35-7.26 (m, 1H), 3.00 ( t, J=7.5Hz, 2H), 2.88 (d, J=11.3Hz, 2H), 2.44 (t, J=7.1Hz, 2H), 2.07 (p, J=7.3Hz, 2H), 1.93 (t, J=11.5Hz, 2H), 1.60 (d, J=12.8Hz, 2H), 1.40–1.26 (m, 1H), 1.21 (td, J=12.2, 3.3Hz, 2H), 0.90 (d, J=6.4 Hz,3H).MS(ESI)m/z322.2([M+H] ⁺ )

Example 8: 3-(3,4-Difluorophenyl)-5-(4-(piperidin-1-yl)butyl)-1,2,4-oxadiazole (8) Substitute piperidine into piperidine, and prepare the target compound according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.94-7.88(m, 1H), 7.85(dd, J=9.6, 4.0Hz, 1H), 7.34-7.24(m, 1H), 3.57(d,J＝11.6Hz,2H),3.02(dt,J＝11.5,6.1Hz,4H),2.64(dd,J＝21.0,9.9Hz,2H),2.32(q,J＝13.0Hz,2H) ), 2.18–2.05 (m, 2H), 2.03–1.92 (m, 3H), 1.92–1.82 (m, 2H), 1.42 (dd, J=25.8, 12.8Hz, 1H). MS (ESI) m/z 322.2([M+H] ⁺ ).

Example 9: 3-(3,4-Difluorophenyl)-5-(3-(4-methylpiperazin-1-yl)propyl)-1,2,4-oxadiazole (9) The target compound was prepared according to the method of Example 1 by replacing 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and replacing 4-methylpiperidine with 4-methylpiperazine.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.89 (t, J=7.4Hz, 1H), 7.85 (dd, J=9.2, 3.2Hz, 1H), 7.36-7.23 (m, 1H), 3.00(t, J＝7.5Hz, 2H), 2.88(d, J＝11.3Hz, 2H), 2.44(t, J＝7.1Hz, 2H), 2.07(p, J＝7.3Hz, 2H) ,1.93(t,J＝11.5Hz,2H),1.60(d,J＝12.8Hz,2H),1.21(td,J＝12.2,3.3Hz,2H), 0.90(d,J＝6.4Hz,3H) .MS(ESI)m/z 323.2([M+H] ⁺ ).

Example 10: 4-(4-(3-(3,4-Difluorophenyl)-1,2,4-oxadiazol-5-yl)butyl)morpholine (10)

The target compound was prepared according to the method of Example 1 by replacing 4-methylpiperidine with morpholine.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.93-7.88 (m, 1H), 7.88-7.81 (m, 1H), 7.33-7.25 (m, 1H), 4.14 (s, 4H) ),3.23(dd,J＝45.6,39.6Hz,2H),3.05(dd,J＝12.1,5.1Hz,4H),2.13(dt,J＝15.7,9.0Hz,2H),2.05–1.95(m, 2H),1.84(s,2H).MS(ESI)m/z 324.2([M+H] ⁺ ).

Example 11: 3-(3,4-Difluorophenyl)-5-(3-(pyrrolidin-1-yl)propyl)-1,2,4-oxadiazole (11) The valeryl chloride was replaced with 4-chlorobutyryl chloride, and the 4-methylpiperidine was replaced with pyrrolidine, and the target compound was prepared according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.89 (t, J=7.4Hz, 1H), 7.85 (dd, J=9.2, 3.2Hz, 1H), 7.36-7.23 (m, 1H), 3.88 (d, J=5.3Hz, 2H), 3.34–3.21 (m, 2H), 3.17 (t, J=7.0Hz, 2H), 2.85 (dd, J=17.5, 7.3Hz, 2H), 2.65–2.49 (m, 2H), 2.28 (dd, J=12.8, 7.7 Hz, 2H), 2.21–2.04 (m, 2H). MS (ESI) m/z 294.1 ([M+H] ⁺ ).

Example 12: 4-(3-(3-(2,4-Dichlorobenzene)-1,2,4-oxadiazol-5-yl)propyl)morpholine (12)

Replace 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with morpholine, according to the method of Example 1 Preparation of the target compound.

Nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.88 (d, J=8.4 Hz, 1H), 7.59 (d, J=1.6 Hz, 1H), 7.41 (dd, J=8.4, 1.8Hz,1H),4.34(t,J=12.3Hz,2H),4.02(d,J=11.6Hz,2H),3.53(d,J=11.7Hz,2H),3.21(dd,J=16.2, 9.4Hz, 4H), 2.93(d, J=10.5Hz, 2H), 2.73–2.52(m, 2H). MS(ESI) m/z 343.1([M+H] ⁺ ).

Example 13: 3-(2,4-Dichlorobenzene)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (13)

Substitute 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile and 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.88 (d, J=8.4 Hz, 1H), 7.58 (d, J=1.4 Hz, 1H), 7.41 (d, J=8.4 Hz) ,1H),3.61(s,2H),3.17(t,J＝6.8Hz,4H),2.60(dt,J＝14.9,11.5Hz,4H),2.02(s,2H),1.93–1.54(m, 3H),1.06(d,J=6.5Hz,3H).MS(ESI)m/z355.1([M+H] ⁺ ).

Example 14: 4-(3-(3-(2,4-Dichlorobenzene)-1,2,4-oxadiazol-5-yl)butyl)morpholine (14)

Substitute 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile and 4-methylpiperidine with morpholine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.90 (d, J=8.4 Hz, 1H), 7.57 (d, J=1.7 Hz, 1H), 7.40 (dd, J=8.4, 1.8Hz,1H),4.32(t,J=12.1Hz,2H),4.00(d,J=12.0Hz,2H),3.49(d,J=11.4Hz,2H),3.08(dd,J=16.4, 9.3Hz, 4H), 2.89(s, 2H), 2.23–2.09(m, 2H), 2.03(dd, J=15.1, 7.5Hz, 2H). MS(ESI) m/z 357.1([M+H] ⁺ ).

Example 15: 3-(2,4-Dichlorobenzene)-5-(4-(4-methylpiperidin-1-yl)butyl)-1,2,4-oxadiazole (15)

Replace 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile, and prepare the target compound according to the method of Example 1

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.91 (d, J=8.4 Hz, 1H), 7.58 (d, J=1.6 Hz, 1H), 7.40 (dd, J=8.4, 1.7Hz, 1H), 3.58(d, J＝11.8Hz, 2H), 3.08(t, J＝7.2Hz, 2H), 3.03–2.94(m, 2H), 2.62(dd, J＝21.1, 10.4Hz, 2H), 2.22–2.08 (m, 2H), 2.08–1.93 (m, 4H), 1.83 (d, J=14.4Hz, 2H), 1.66–1.52 (m, 1H), 1.06 (d, J=6.4Hz) ,3H).MS(ESI)m/z369.1 ([M+H] ⁺ ).

Example 16: 3-(2,4-Dichlorobenzene)-5-(4-(piperidin-1-yl)butyl)-1,2,4-oxadiazole (16)

Substitute 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile and 4-methylpiperidine with piperidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.91 (d, J=8.4 Hz, 1H), 7.58 (d, J=1.8 Hz, 1H), 7.40 (dd, J=8.4, 1.9Hz, 1H), 3.57(d, J＝11.7Hz, 2H), 3.08(t, J＝7.2Hz, 2H), 3.04–2.93(m, 2H), 2.63(dd, J＝21.5, 9.7Hz, 2H), 2.33 (dd, J=27.0, 12.9Hz, 2H), 2.22–2.08 (m, 2H), 1.95 (ddd, J=43.1, 22.2, 11.0Hz, 5H), 1.43 (dt, J=25.7, 8.3Hz,1H).MS(ESI)m/z 355.2 ([M+H] ⁺ ).

Example 17: 3-(2,4-Dichlorobenzene)-5-(4-(pyrrolidin-1-yl)butyl)-1,2,4-oxadiazole (17)

Substitute 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile and 4-methylpiperidine with pyrrolidine, and prepare the target compound according to the method of Example 1.

Nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.90 (d, J=8.4 Hz, 1H), 7.56 (s, 1H), 7.39 (d, J=8.4 Hz, 1H), 3.84 (d, J=5.3Hz, 2H), 3.18 (dd, J=15.3, 6.2Hz, 2H), 3.07 (t, J=7.1Hz, 2H), 2.94–2.78 (m, 2H), 2.32– 2.18 ( m,2H),2.18–2.06(m,4H),2.06–1.94(m,2H).MS(ESI)m/z 341.2 ([M+H] ⁺ ).

Example 18: 3-(3,5-Dichlorobenzene)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (18)

Substitute 3,4-difluorobenzonitrile with 3,5-dichlorobenzonitrile, and replace 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.96 (s, 2H), 7.52 (s, 1H), 3.64 (d, J=11.7 Hz, 2H), 3.15 (dd, J= 13.6, 6.7Hz, 4H), 2.69 (dd, J=21.7, 10.4Hz, 2H), 2.64–2.51 (m, 2H), 2.06 (dd, J=24.7, 11.8Hz, 2H), 1.86 (d, J =14.2Hz,2H),1.65(d,J=6.2Hz,1H),1.08(t,J=8.5Hz,3H).MS (ESI)m/z 355.1([M+H] ⁺ )

Example 19: 4-(3-(3-(3,5-Dichlorobenzene)-1,2,4-oxadiazol-5-yl)propyl)morpholine (19)

Replace 3,4-difluorobenzonitrile with 3,5-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with morpholine, according to the method of Example 1 Preparation of target compounds

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.95(s,2H),7.51(s,1H),4.33(t,J=12.3Hz,2H),4.03(d,J= 11.9Hz, 2H), 3.56 (d, J=11.8Hz, 2H), 3.30–3.13 (m, 4H), 2.97 (d, J=10.0Hz, 2H), 2.61 (dd, J=15.3, 7.4Hz, 2H).MS(ESI)m/z 343.1([M+H] ⁺ ).

Example 20: 4-(4-(3-(3-Chloro-4-fluorobenzene)-1,2,4-oxadiazol-5-yl)butyl)morpholine (20)

Substitute 3,4-difluorobenzonitrile with 3-chloro-4-fluorobenzonitrile and 4-methylpiperidine with morpholine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.15 (d, J=7.0 Hz, 1H), 8.01-7.93 (m, 1H), 7.28 (d, J=4.0 Hz, 1H) ,4.33(t,J＝12.2Hz,2H), 4.01(d,J＝12.3Hz,2H),3.50(d,J＝11.7Hz,2H),3.06(t,J＝7.0Hz,4H), 2.90 (s,2H),2.15(dt,J=15.8,9.0Hz,2H),2.06–1.94(m,2H).MS(ESI) m/z 341.2([M+H] ⁺ ).

Example 21: 3-(3-Chloro-4-fluorobenzene)-5-(4-(piperidin-1-yl)butyl)-1,2,4-oxadiazole (12)

Substitute 3,4-difluorobenzonitrile with 3-chloro-4-fluorobenzonitrile and 4-methylpiperidine with piperidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectral data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.14 (t, J=7.4Hz, 1H), 8.01-7.92 (m, 1H), 7.26 (dd, J=14.6, 6.0Hz, 1H), 3.56(d, J＝11.6Hz, 2H), 3.10–2.95(m, 4H), 2.65(dd, J＝21.7, 9.8Hz, 2H), 2.30(dd, J＝26.4, 13.2Hz, 2H) ),2.20–2.04(m,2H),1.96(td,J=14.8,7.4Hz,2H),1.91–1.66(m,3H),1.43(dt,J=25.2,7.9Hz,1H).MS( ESI)m/z339.2([M+H] ⁺ ).

Example 22: 3-(3-Chloro-4-fluorobenzene)-5-(4-(4-methylpiperidin-1-yl)butyl)-1,2,4-oxadiazole (22)

3,4-difluorobenzonitrile was replaced with 3-chloro-4-fluorobenzonitrile, and the target compound was prepared according to the method of Example 1

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.14 (d, J=4.9 Hz, 1H), 8.04-7.92 (m, 1H), 7.37-7.22 (m, 1H), 3.58 ( d, J=11.5Hz, 2H), 3.19–2.93 (m, 4H), 2.64 (dd, J=22.3, 10.3Hz, 2H), 2.28–2.08 (m, 4H), 2.08–1.92 (m, 4H) ,1.83(d,J=14.1Hz,1H),1.24–0.98(m,3H).MS(ESI)m/z 352.2([M+H] ⁺ ).

Example 23: 3-(3-Chloro-4-fluorobenzene)-5-(4-(4-methylpiperazin-1-yl)butyl)-1,2,4-oxadiazole (23)

3,4-difluorobenzonitrile was replaced with 3-chloro-4-fluorobenzonitrile, 4-methylpiperidine was replaced with 4-methylpiperazine, and the target compound was prepared according to the method of Example 1

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.14 (d, J=6.3 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 6.85 (d, J=8.4 Hz) ,1H),2.99(t,J＝7.5Hz,2H),2.50(s,6H),2.45–2.38(m,4H),2.31(s,3H),1.98–1.87(m,2H),1.71– 1.59(m,2H).MS(ESI)m/z 353.1([M+H] ⁺ ).

Example 24: 4-(3-(3-(3-Chloro-4-fluorobenzene)-1,2,4-oxadiazol-5-yl)propyl)morpholine (24)

3,4-difluorobenzonitrile was replaced by 3-chloro-4-fluorobenzonitrile, 5-chlorovaleryl chloride was replaced by 4-chlorobutyryl chloride, and 4-methylpiperidine was replaced by morpholine, as described in Example 1. method to prepare the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.15 (d, J=7.0 Hz, 1H), 7.98 (dd, J=7.7, 5.3 Hz, 1H), 7.33-7.24 (m, 1H), 4.33(d, J＝11.8 Hz, 2H), 4.03(dd, J＝10.0, 5.2Hz, 2H), 3.57(d, J＝10.7Hz, 2H), 3.24(d, J＝7.5Hz, 2H), 3.18(t, J=7.0Hz, 2H), 2.95(s, 2H), 2.68–2.53(m, 2H). MS (ESI) m/z 326.1([M+H] ⁺ ).

Example 25: 3-(3-Chloro-4-fluorobenzene)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (25)

Substitute 3,4-difluorobenzonitrile with 3-chloro-4-fluorobenzonitrile, and replace 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and prepare the target compound according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ8.14 (dd, J=7.0, 1.9Hz, 1H), 8.01-7.93 (m, 1H), 7.32-7.23 (m, 1H), 3.64(d,J＝11.9Hz,2H), 3.14(t,J＝6.9Hz,4H),2.77-2.63(m,2H),2.59(dd,J＝15.4,7.6Hz,2H), 2.14-1.98 (m,2H),1.86(d,J＝14.3Hz,2H),1.58(d,J＝73.3Hz,1H),1.06(t, J＝8.7Hz,3H).MS(ESI)m/z 338.1 ([M+H] ⁺ ).

Example 26: 3-(3-Chloro-4-fluorobenzene)-5-(4-(pyrrolidin-1-yl)butyl)-1,2,4-oxadiazole (26)

Substitute 3,4-difluorobenzonitrile with 3-chloro-4-fluorobenzonitrile and 4-methylpiperidine with pyrrolidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.15 (dd, J=7.0, 1.8 Hz, 1H), 7.80 (d, J=8.6 Hz, 1H), 6.85 (d, J= 8.7Hz,1H),4.05(s,2H),3.88(s,2H),3.55(t,J=6.4Hz,2H),3.19(s,2H),3.00(dt,J=12.4,6.2Hz, 2H), 2.82(s, 2H), 2.14(d, J=21.0Hz, 2H), 1.99(dd, J=9.1, 4.8Hz, 2H). MS(ESI) m/z 324.1([M+H] ⁺ ).

Example 27: 3-(3-Chloro-4-fluorobenzene)-5-(3-(pyrrolidin-1-yl)propyl)-1,2,4-oxadiazole (27)

3,4-difluorobenzonitrile was replaced by 3-chloro-4-fluorobenzonitrile, 5-chlorovaleryl chloride was replaced by 4-chlorobutyryl chloride, and 4-methylpiperidine was replaced by pyrrolidine, as described in Example 1. method to prepare the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.14 (d, J=6.3 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 6.85 (d, J=8.4 Hz) ,1H),3.55(s,2H),3.32(s,2H),3.07(d,J＝15.9Hz,2H),2.39(s,2H),2.15(s,4H),1.99(s,2H) .MS (ESI) m/z 310.1([M+H] ⁺ ).

Example 28: 3-(3-Chloro-4-fluorobenzene)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (28)

Change 3,4-difluorobenzonitrile to 3-chloro-4-fluorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride, 4-methylpiperidine to 4-methylpiperazine, press The method of Example 1 prepares the target compound.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ8.14 (dd, J=7.0, 1.9Hz, 1H), 8.01-7.93 (m, 1H), 7.32-7.23 (m, 1H), 3.03(t,J＝7.5Hz,2H), 2.88(d,J＝7.2Hz,2H),2.44(t,J＝7.1Hz,2H),2.07(p,J＝7.3Hz,2H),1.93 ( t,J＝11.5Hz,2H),1.60(d,J＝12.8Hz,2H),1.21(td,J＝12.2,3.3Hz,2H), 0.90(d,J＝6.4Hz,3H).MS( ESI) m/z 339.1([M+H] ⁺ ).

Example 29: 3-(2,5-Dichlorobenzene)-5-(4-(4-methylpiperidin-1-yl)butyl)-1,2,4-oxadiazole (29)

Substitute 3,4-difluorobenzonitrile with 2,5-dichlorobenzonitrile, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.95 (d, J=2.4 Hz, 1H), 7.50 (d, J=8.6 Hz, 1H), 7.43 (dd, J=8.6, 2.3Hz, 1H), 3.59(d, J=11.7Hz, 2H), 3.09(t, J=7.2Hz, 2H), 3.00(s, 2H), 2.69–2.53(m, 2H), 2.24–2.08( m,2H),2.08–1.94(m,5H),1.83(d,J=14.1Hz,2H),1.07(d,J=6.4Hz,3H).MS(ESI)m/z 369.1([M+ H] ⁺ ).

Example 30: 4-(4-(3-(2,3-Dichlorobenzene)-1,2,4-oxadiazol-5-yl)butyl)morpholine (30)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile and 4-methylpiperidine with morpholine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.78 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.9 Hz) ,1H),4.31(t,J＝12.2Hz,2H),3.99(dd,J＝12.9,2.9Hz,2H),3.49(d,J＝11.9Hz,2H),3.07(dt,J＝12.2, 6.0Hz,4H), 2.97–2.83(m,2H), 2.23–2.07(m,2H), 2.07–1.92(m,2H). MS(ESI) m/z 357.1([M+H] ⁺ ).

Example 31: 3-(2,3-Dichlorobenzene)-5-(4-(4-methylpiperidin-1-yl)butyl)-1,2,4-oxadiazole (31)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.79 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.9 Hz) ,1H),3.58(d,J＝11.5Hz,2H),3.07(q,J＝7.6Hz,2H),3.04–2.93(m,2H),2.63(q,J＝10.2Hz,2H), 2.15 (ddd, J=11.5, 10.1, 6.2Hz, 2H), 2.10–1.91 (m, 4H), 1.82 (d, J=13.3Hz, 2H), 1.63 (dd, J=10.6, 7.5Hz, 1H), 1.06(t,J=6.0Hz,3H).MS(ESI)m/z 369.1([M+H] ⁺ ).

Example 32: 3-(2,5-Dichlorobenzene)-5-(4-(piperidin-1-yl)butyl)-1,2,4-oxadiazole (32)

Substitute 3,4-difluorobenzonitrile with 2,5-dichlorobenzonitrile and 4-methylpiperidine with piperidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.94 (d, J=2.0 Hz, 1H), 7.48 (d, J=8.6 Hz, 1H), 7.42 (dd, J=8.6, 2.0Hz, 1H), 3.57(d, J= 11.6Hz, 2H), 3.08(t, J=7.2Hz, 2H), 3.00(dt, J=9.2, 5.3Hz, 2H), 2.64(dd, J= 21.7, 9.9Hz, 2H), 2.32 (dd, J=26.8, 12.9Hz, 2H), 2.20–2.08 (m, 2H), 2.06–1.96 (m, 2H), 1.96–1.81 (m, 3H), 1.43 (dt,J=25.5,8.1Hz,1H).MS(ESI) m/z 355.1([M+H] ⁺ ).

Example 33: 3-(2,3-Dichlorobenzene)-5-(4-(piperidin-1-yl)butyl)-1,2,4-oxadiazole (33)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile and 4-methylpiperidine with piperidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.79 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.9 Hz) ,1H),3.56(d,J＝11.4Hz,2H),3.08(t,J＝7.2Hz,2H),3.00(dt,J＝9.3,5.3Hz,2H),2.64(dd,J＝21.7, 9.9Hz, 2H), 2.32 (dd, J=26.8, 12.9Hz, 2H), 2.22–2.08 (m, 2H), 2.01 (dd, J= 15.1, 7.5Hz, 2H), 1.97–1.81 (m, 3H) ),1.42(dt,J=25.6,8.1Hz,1H).MS(ESI)m/z 355.1([M+H] ⁺ ).

Example 34: 3-(2,3-Dichlorobenzene)-5-(4-(pyrrolidin-1-yl)butyl)-1,2,4-oxadiazole (34)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile and 4-methylpiperidine with pyrrolidine, and prepare the target compound according to the method of Example 1.

Nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.79 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.36 (t, J=7.9 Hz) ,1H),4.94(s,2H),3.84(s,2H),3.19(s,2H),3.05(t,J＝6.0Hz,2H),2.87(s,2H),2.13(s,4H) ,1.98(s, 2H).MS(ESI)m/z 341.1([M+H] ⁺ ).

Example 35: 3-(2,3-Dichlorobenzene)-5-(4-(4-methylpiperazin-1-yl)butyl)-1,2,4-oxadiazole (35)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile and 4-methylpiperidine with 4-methylpiperazine, and prepare the target compound according to the method of Example 1.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.79 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.9 Hz, 1H), 3.00 (t , J=7.5Hz, 2H), 2.50 (s, 6H), 2.42–2.36 (m, 4H), 2.31 (s, 3H), 1.98–1.87 (m, 2H), 1.71–1.59 (m, 2H). MS(ESI) m/z 370.1([M+H] ⁺ ).

Example 36: 3-(2,5-Dichlorobenzene)-5-(3-(piperidin-1-yl)propyl)-1,2,4-oxadiazole (36)

Replace 3,4-difluorobenzonitrile with 2,5-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with piperidine, as in Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.92 (d, J=2.4 Hz, 1H), 7.49 (d, J=8.6 Hz, 1H), 7.43 (dd, J=8.6, 2.4Hz, 1H), 3.62 (d, J = 11.6Hz, 2H), 3.27–3.08 (m, 4H), 2.78–2.50 (m, 4H), 2.43–2.19 (m, 2H), 1.92 (t, J =17.8Hz,3H),1.53–1.37(m,1H).MS(ESI)m/z 341.1([M+H] ⁺ ).

Example 37: 3-(2,5-Dichlorobenzene)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (37)

Substitute 3,4-difluorobenzonitrile with 2,5-dichlorobenzonitrile, and replace 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.94 (d, J=2.0 Hz, 1H), 7.48 (d, J=8.6 Hz, 1H), 7.42 (dd, J=8.6, 2.0Hz, 1H), 3.64 (d, J=11.9Hz, 2H), 3.14 (t, J=6.9Hz, 4H), 2.77–2.63 (m, 2H), 2.59 (dd, J=15.4, 7.6Hz, 2H),2.14–1.98(m,2H),1.86(d,J=14.3Hz,2H),1.58(m,1H),1.06(t,J=8.7Hz,3H).MS(ESI)m/z 355.1([M+H] ⁺ ).

Example 38: 3-(2,3-Dichlorobenzene)-5-(3-(piperidin-1-yl)propyl)-1,2,4-oxadiazole (38)

Replace 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with piperidine, as in Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.00 (d, J=7.9 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.36 (t, J=7.9 Hz) ,1H),3.62(d,J＝11.5Hz,2H),3.14(t,J＝6.9Hz,4H),2.70(dd,J＝20.0,10.0Hz,2H),2.63–2.52(m, 2H) ,2.34(dd,J＝26.1,12.8Hz,2H),2.00–1.73(m,1H),1.45(dd,J＝25.4, 12.6Hz,1H).MS(ESI)m/z 341.1([M+ H] ⁺ ).

Example 39: 3-(2,3-Dichlorobenzene)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (39) 3,4-difluorobenzonitrile was replaced with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride was replaced with 4-chlorobutyryl chloride, and the target compound was prepared according to the method of Example 1.

Nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.99 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.36 (t, J=7.9 Hz) ,1H),3.64(d,J＝11.9Hz,2H),3.14(t,J＝6.9Hz,4H),2.77–2.63(m,2H),2.59(dd,J＝15.4,7.6Hz,2H) ,2.14–1.98(m,2H),1.86(d,J＝14.3Hz,2H),1.58(m,1H),1.06(t,J＝8.7Hz,3H).MS(ESI)m/z 355.21( [M+H] ⁺ ).

Example 40: 3-(2,3-Dichlorobenzene)-5-(3-(pyrrolidin-1-yl)propyl)-1,2,4-oxadiazole (40)

Replace 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with pyrrolidine, as in Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.79 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.9 Hz) ,1H),3.88(s,2H),3.31– 3.20(m,2H),3.15(t,J=6.9Hz,2H),2.85(s,2H),2.61–2.50(m,2H),2.28( s, 2H),2.12(s,2H).MS(ESI)m/z 327.1([M+H] ⁺ ).

Example 41: 4-(3-(3-(2,3-Dichlorobenzene)-1,2,4-oxadiazol-5-yl)propyl)morpholine (41)

Replace 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with morpholine, according to the method of Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.79 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.9 Hz) ,1H),4.33(d,J＝11.8Hz,2H),4.03–3.93(m,2H),3.57(d,J＝10.7Hz,2H),3.24(d,J＝7.5Hz,2H), 3.18 (t, J=7.0Hz, 2H), 2.95(s, 2H), 2.68–2.53(m, 2H). MS(ESI) m/z 343.1 ([M+H] ⁺ ).

Example 42: 3-(3,5-Dichlorobenzene)-5-(4-(piperidin-1-yl)butyl)-1,2,4-oxadiazole (42)

Substitute 3,4-difluorobenzonitrile with 3,5-dichlorobenzonitrile and 4-methylpiperidine with piperidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.96(s,2H),7.52(s,1H),3.57(d,J=11.7Hz,2H),3.08(t,J= 7.2Hz, 2H), 3.04–2.93 (m, 2H), 2.63 (dd, J=21.5, 9.7Hz, 2H), 2.33 (dd, J=27.0, 12.9Hz, 2H), 2.22–2.08 (m, 2H ),1.95(ddd,J=43.1,22.2,11.0Hz,5H),1.43(dt,J=25.7,8.3Hz,1H). MS(ESI)m/z 355.1([M+H] ⁺ ).

Example 43: 3-(3,5-Dichlorobenzene)-5-(4-(pyrrolidin-1-yl)butyl)-1,2,4-oxadiazole (43)

Substitute 3,4-difluorobenzonitrile with 3,5-dichlorobenzonitrile and 4-methylpiperidine with pyrrolidine, and prepare the target compound according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.85(s,2H),7.54(s,1H),3.83(d,J=5.1Hz,2H),3.15-3.08(m, 2H), 3.05 (t, J=7.1Hz, 2H), 2.79 (dd, J=17.6, 7.4Hz, 2H), 2.34–2.20 (m, 2H), 2.17–2.04 (m, 4H), 2.04– 1.96 (m,2H).MS(ESI)m/z 341.1([M+H] ⁺ ).

Example 44: 4-(4-(3-(3,5-Dichlorobenzene)-1,2,4-oxadiazol-5-yl)butyl)morpholine (44)

3,4-difluorobenzonitrile was replaced with 3,5-dichlorobenzonitrile, 4-methylpiperidine was replaced with morpholine, and the target compound was prepared according to the method of Example 1,

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ8.00(s,2H),7.48(s,1H),4.32(t,J=12.1Hz,2H),4.00(d,J= 12.0Hz, 2H), 3.49(d, J＝11.4Hz, 2H), 3.08(dd, J＝16.4, 9.3Hz, 4H), 2.89(s, 2H), 2.23–2.09(m, 2H), 2.03( dd, J＝15.1,7.5Hz,2H).MS(ESI)m/z357.1([M+H] ⁺ ).

Example 45: 3-(3,5-Dichlorobenzene)-5-(4-(4-methylpiperidin-1-yl)butyl)-1,2,4-oxadiazole (45)

Substitute 3,4-difluorobenzonitrile with 3,5-dichlorobenzonitrile, and prepare the target compound according to the method of Example 1.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.94(s, 2H), 7.52 (s, 1H), 2.99(t, J=7.5Hz, 2H), 2.50(s, 6H) ,2.45–2.38(m,4H),2.31(s,3H), 1.98–1.87(m,2H),1.71–1.59(m,2H).MS(ESI)m/z 369.1([M+H] ⁺ ).

Example 46: 3-(3,5-Dichlorobenzene)-5-(3-(pyrrolidin-1-yl)propyl)-1,2,4-oxadiazole (46)

Replace 3,4-difluorobenzonitrile with 3,5-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with pyrrolidine, as in Example 1 Preparation of the target compound.

NMR data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.86(s,2H),7.57(s,1H),3.88(d,J=5.3Hz,2H),3.34-3.21(m, 2H), 3.17(t, J＝7.0Hz, 2H), 2.85(dd, J＝17.5, 7.3Hz, 2H), 2.65–2.49(m, 2H), 2.28(dd, J＝12.8, 7.7Hz, 2H) ),2.21–2.04(m,2H).MS(ESI)m/z327.1([M+H] ⁺ ).

Example 47: 3-(3,5-Dichlorobenzene)-5-(3-(piperidin-1-yl)propyl)-1,2,4-oxadiazole (47)

Replace 3,4-difluorobenzonitrile with 3,5-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with piperidine, according to the method of Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ )δ7.96(s,2H),7.52(s,1H),3.62(d,J=11.5Hz,2H),3.14(t,J= 6.9Hz, 4H), 2.70 (dd, J=20.0, 10.0Hz, 2H), 2.63–2.52 (m, 2H), 2.34 (dd, J=26.1, 12.8Hz, 2H), 2.00–1.73 (m, 1H ),1.45(dd,J=25.4,12.6Hz,1H).MS(ESI)m/z 341.1([M+H] ⁺ ).

Example 48: 3-(2,4-Dichlorobenzene)-5-(3-(piperidin-1-yl)propyl)-1,2,4-oxadiazole (48)

Replace 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with piperidine, according to the method of Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.90 (d, J=8.4 Hz, 1H), 7.57 (d, J=1.7 Hz, 1H), 7.40 (dd, J=8.4, 1.8Hz, 1H), 3.62 (d, J=11.5Hz, 2H), 3.14 (t, J=6.9Hz, 4H), 2.70 (dd, J=20.0, 10.0Hz, 2H), 2.63– 2.52 (m, 2H),2.34(dd,J=26.1,12.8Hz,2H),2.00–1.73(m,2H),1.45(dd,J=25.4,12.6Hz,2H).MS(ESI)m/z 341.1([ M+H] ⁺ ).

Example 49: 3-(2,4-Dichlorobenzene)-5-(3-(pyrrolidin-1-yl)propyl)-1,2,4-oxadiazole (49)

Replace 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with pyrrolidine, as in Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.91 (d, J=8.4 Hz, 1H), 7.58 (d, J=1.6 Hz, 1H), 7.40 (dd, J=8.4, 1.7Hz, 1H), 3.88 (d, J=5.3 Hz, 2H), 3.34–3.21 (m, 2H), 3.17 (t, J=7.0Hz, 2H), 2.85 (dd, J=17.5, 7.3Hz, 2H), 2.65–2.49(m, 2H), 2.28(dd, J=12.8, 7.7Hz, 2H), 2.21–2.04(m, 2H). MS(ESI) m/z 327.1([M+H] ⁺ ).

Example 50: 3-(2,4-Dichlorobenzene)-5-(3-(4-methylpiperazin-1-yl)propyl)-1,2,4-oxadiazole (50)

Replace 3,4-difluorobenzonitrile with 2,4-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with 4-methylpiperazine, as per implementation The title compound was prepared by the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.90 (d, J=8.4 Hz, 1H), 7.56 (s, 1H), 7.39 (d, J=8.4 Hz, 1H), 3.00 (t, J＝7.5Hz, 2H), 2.88(d, J＝11.3Hz, 2H), 2.44(t, J＝7.1Hz, 2H), 2.07(p, J＝7.3Hz, 2H), 1.93(t , J＝11.5Hz, 2H), 1.60(d, J＝12.8Hz, 2H), 1.21(td, J＝12.2, 3.3Hz, 2H), 0.90(s, 3H). MS(ESI) m/z 356.1 ([M+H] ⁺ ).

Example 51: 3-(2,3-Dichlorobenzene)-5-(3-(4-methylpiperazin-1-yl)propyl)-1,2,4-oxadiazole (51)

Replace 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with 4-methylpiperazine. The title compound was prepared by the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.79 (d, J=7.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.9 Hz) ,1H),2.53(t,J＝7.5Hz,2H),2.38(d,J＝7.2Hz,2H),2.29(m,4H),1.93(t,J＝7.8Hz,2H),2.14(s , 3H).MS(ESI)m/z 356.1([M+H] ⁺ ).

Example 52: 3-(3,4-Dichlorobenzene)-5-(3-(piperidin-1-yl)propyl)-1,2,4-oxadiazole (52)

Replace 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with piperidine, as in Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.00 (d, J=6.3 Hz, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.48 (d, J=8.4 Hz) ,1H),3.62(d,J＝11.5Hz,2H),3.14(t,J＝6.9Hz,4H),2.70(dd,J＝20.0,10.0Hz,2H),2.63–2.52(m, 2H) ,2.34(dd,J＝26.1,12.8Hz,2H),2.00–1.73(m,2H),1.45(dd,J＝25.4, 12.6Hz,2H).MS(ESI)m/z 341.1([M+ H] ⁺ ).

Example 53: 4-(3-(3-(3,4-Dichlorobenzene)-1,2,4-oxadiazol-5-yl)propyl)morpholine (53)

Replace 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with morpholine, according to the method of Example 1 Preparation of the target compound.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.00 (d, J=6.3 Hz, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.48 (d, J=8.4 Hz, 1H), 4.33 (d , J＝11.8Hz, 2H), 4.03–3.93(m, 2H), 3.57(d, J＝10.7Hz, 2H), 3.24(d, J＝7.5Hz, 2H), 3.18(t, J＝7.0Hz ,2H),2.95(s,2H),2.68–2.53(m,2H).MS(ESI)m/z343.1([M+H] ⁺ ).

Example 54: 3-(3,4-Dichlorobenzene)-5-(4-(piperidin-1-yl)butyl)-1,2,4-oxadiazole (54)

Substitute 3,4-difluorobenzonitrile with 3,4-dichlorobenzonitrile and 4-methylpiperidine with piperidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.01 (d, J=6.3 Hz, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.48 (d, J=8.4 Hz) ,1H),3.57(d,J＝11.7Hz,2H),3.08(t,J＝7.2Hz,2H),3.04–2.93(m,2H),2.63(dd,J＝21.5,9.7Hz,2H) , 2.33(dd,J＝27.0,12.9Hz,2H),2.22–2.08(m,2H),1.95(ddd,J＝43.1,22.2,11.0Hz,5H),1.43(dt,J＝25.7,8.3Hz ,1H).MS(ESI)m/z 355.1([M+H] ⁺ ).

Example 55: 3-(3,4-Dichlorobenzene)-5-(3-(pyrrolidin-1-yl)propyl)-1,2,4-oxadiazole (55)

Replace 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and 4-methylpiperidine with pyrrolidine, as in Example 1 Preparation of the target compound.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.95 (d, J=6.3 Hz, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.48 (d, J=8.4 Hz) ,1H),3.88(d,J＝5.3Hz,2H),3.34–3.21(m,2H),3.17(t,J＝7.0Hz,2H),2.85(dd,J＝17.5,7.3Hz,2H) , 2.65–2.49(m, 2H), 2.28(dd, J=12.8, 7.7Hz, 2H), 2.21–2.04(m, 2H). MS(ESI) m/z 327.1([M+H] ⁺ ).

Example 56: 3-(3,4-Dichlorobenzene)-5-(4-(pyrrolidin-1-yl)butyl)-1,2,4-oxadiazole (56)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile and 4-methylpiperidine with pyrrolidine, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.00 (d, J=6.3 Hz, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.48 (d, J=8.4 Hz) ,1H),3.83(d,J＝5.1Hz,2H),3.15–3.08(m,2H),3.05(t,J＝7.1Hz,2H),2.79(dd,J＝17.6,7.4Hz,2H) ,2.34–2.20(m,2H),2.17–2.04(m,4H),2.04–1.96(m,2H).MS(ESI) m/z 341.1([M+H] ⁺ ).

Example 57: 3-(3,4-Dichlorobenzene)-5-(3-(4-methylpiperidin-1-yl)propyl)-1,2,4-oxadiazole (57)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, and replace 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.92 (d, J=6.3 Hz, 1H), 7.90 (d, J=8.3 Hz, 1H), 7.48 (d, J=8.3 Hz) ,1H),3.00(t,J＝7.5Hz,2H),2.88(d,J＝11.3Hz,2H),2.44(t,J＝7.1Hz,2H),2.07(p,J＝7.3Hz,2H) ), 1.93(t, J＝11.5Hz, 2H), 1.60(d, J＝12.8Hz, 2H), 1.40–1.26(m, 1H), 1.21 (td, J＝12.2, 3.3Hz, 2H), 0.90 (d,J=6.4Hz,3H).MS(ESI)m/z 355.1 ([M+H] ⁺ ).

Example 58: 3-(3,4-Dichlorobenzene)-5-(4-(4-methylpiperidin-1-yl)butyl)-1,2,4-oxadiazole (58)

Substitute 3,4-difluorobenzonitrile with 2,3-dichlorobenzonitrile, and prepare the target compound according to the method of Example 1.

The nuclear magnetic data and mass spectrometry data are: ¹ H NMR (400MHz, CDCl ₃ ) δ 7.89 (d, J=6.4 Hz, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.48 (d, J=8.4 Hz) ,1H),2.99(t,J＝7.5Hz,2H),2.50(s,6H),2.22-2.08(m,2H),2.08-1.93(m,4H),1.83(d,J＝14.4Hz, 2H),1.66–1.52(m,1H),1.06(d,J=6.4Hz,3H).MS(ESI)m/z 369.1 ([M+H] ⁺ )

The preferred compound number and its structural formula prepared by the embodiment of table 1

Second Aspect: Examples of Pharmacological Aspects

Example 59: Preparation of σ ₁ Receptor Membrane and Determination of Ligand Affinity (Inhibition Rate)

Preparation of σ ₁ Receptor Membranes

The guinea pigs were decapitated, operated on ice, the brains were quickly removed, the tissues were combined into a centrifuge tube, 0.01M Tris HCl + 0.32M sucrose solution was added to the 4th gear for 3-4s, and the homogenization was performed 4 times, and then 0.01M was added. Tris HCl+0.32M sucrose solution was adjusted to 10ml/g, the weight of the homogenized test tube was adjusted with a balance, and centrifuged at 1000r for 10min; Centrifuge at 4°C for 10min; take the supernatant, centrifuge at 11500r for 25min at 4°C; take the precipitate and add 0.01M Tris HCl+0.32M sucrose solution to adjust to 3ml/g, incubate at 25°C for 15min, centrifuge at 11500r for 25min at 4°C, and put the pellet in Store at -80°C for later use.

It includes the following steps: 1. Quantitative determination of protein by Bradford method, referring to the kit instructions.

2. Preparation of homogenate

A: 0.01M Tris-HCl buffer containing 0.32M sucrose solution, pH 7.4.

B: 0.01M Tris-HCl buffer, pH 7.4.

3. Receptor saturation binding experiment.

(1) using an appropriate amount of homogenate for the prepared membrane, disperse it evenly with a homogenizer, add an appropriate amount of the homogenate to reference the protein determination of the quantitative membrane suspension, for subsequent use;

(2) 100 μL of membrane preparation was added to each reaction tube;

(3) Add 100 μL B solution to the total binding tube (TB), and add 100 μL haloperidol (final concentration 10-5M) to the non-specific binding tube (NB);

(4) 10 μL of the radioligand [3H]-(+)-pentazocine was added to each reaction tube, and the final concentrations were 32.00, 16.00, 8.00, 4.00, 2.00, 1.00, 0.50, and 0.25nM in turn;

(5) Incubate each reaction tube at 25°C for 3 hours. After the reaction is completed, the bound ligands are quickly filtered under reduced pressure, washed with ice-cold test buffer, and the filter is taken out into a 2ml scintillation cup, and 1ml of toluene is added. scintillation fluid and mix;

(6) Put the scintillation vial into the liquid scintillation counter for counting.

4. Competitive binding assay of σ ₁ receptor

(1) first use an appropriate amount of homogenate for the prepared membrane, disperse it evenly with a homogenizer, add an appropriate amount of the homogenate to be the suspension of 50ml membrane, for subsequent use;

(2) 100 μL of membrane preparation was added to each reaction tube;

(3) Add 100 μL of B solution to the total binding tube (TB), add 100 μL of haloperidol (final concentration 10-5M) to the non-specific binding tube (NB), and add 100 μL of the test compound to the specific binding tube (SB) of each test compound. Compound (final concentration 10-5M);

(4) Add 10 μL of radioligand [3H]-(+)-pentazocine (final concentration 4nM) to each reaction tube;

(5) Incubate each reaction tube at 25°C for 3 hours. After the reaction is completed, the bound ligands are quickly filtered under reduced pressure. Whatman test paper is saturated with 0.25% PEI solution 2 hours in advance, and fully washed with ice-cold test buffer, and the filter is taken out. Put it into a 2ml scintillation cup, add 1ml of toluene scintillation fluid and mix well;

(6) Put the scintillation vial into the liquid scintillation counter for counting.

5. Data Statistical Processing

TB: Summary and Constants

NB: nonspecific binding constant

SB: Binding constant of the compound

Inhibition rate (I%)=(TB-SB)/(TB-NB)×100%

Example 60: Preparation of σ ₂ Receptor Membrane and Determination of Ligand Affinity (Inhibition Rate)

Preparation of σ ₂ receptor membrane: guinea pigs were decapitated, operated on ice, brains were quickly removed, the tissues were combined into a centrifuge tube, 0.01M Tris HCl + 0.32M sucrose solution was added to homogenize in the fourth gear for 3-4s, and the cells were homogenized. slurry 4 times, then add 0.01M Tris HCl+0.32M sucrose solution to adjust to 10ml/g, adjust the weight of the homogenized test tube with a balance, centrifuge at 1000r for 10min; take the supernatant and add 0.01M Tris HCl+0.32M sucrose solution Adjust to 2ml/g, 1000r, centrifuge at 4°C for 10min; take the supernatant, centrifuge at 11000r, 4°C for 30min; take the precipitate and suspend it in 0.01M Tris HCl+0.32M sucrose solution for 30s, adjust to 3ml/g, incubate at 25°C 15min, centrifuge at 11000g for 30min to take the supernatant, store at -20°C for more than 12h, and incubate with 50Mm-Tris when using.

It includes the following steps: 1. Quantitative determination of protein by Bradford method, referring to the kit instructions.

2. Competitive binding experiment of sigma-2 receptor.

(1) First, use an appropriate amount of homogenate (50mM Tris buffer, pH 7.4) for the prepared membrane, disperse it evenly with a homogenizer, and use it for later use;

(2) 100 μL of membrane preparation and 100 μL of homogenate were added to each reaction tube;

(3) Add 100 μL of homogenate to the total binding tube (TB), add 100 μL of 5uM DTG (final concentration 0.5*10-5M) to the non-specific binding tube (NB), and add 100 μL of the test compound to the specific binding tube (SB) of each test compound. Compound (final concentration 10-5M); 100nM (+)--NANM shields sigma-1 receptor;

(4) 10 μL of radioligand 3H-DTG (final concentration 5nM) was added to each reaction tube (each reaction tube was set with 2 parallel tubes, and each tube was placed on ice when adding samples);

(5) Incubate each reaction tube at 25°C for 120min. After the reaction is completed, the bound ligands are quickly filtered under reduced pressure. Whatman test paper is soaked in 0.5% PEI, washed with ice-cold test buffer, and the filter is taken out and placed in 2ml In the scintillation cup, add 1ml of toluene scintillation fluid and mix well;

(6) Put the scintillation vial into the liquid scintillation counter for counting.

5. Data Statistical Processing

TB: Summary and Constants

NB: nonspecific binding constant

SB: Binding constant of the compound

Inhibition rate (I%)=(TB-SB)/(TB-NB)×100%

Statistical processing of data: The affinity of the compound for σ ₁ receptor and σ ₂ receptor at a concentration of 10 ^-5 M was calculated to obtain the inhibition rate (%). The results are shown in Table 2.

Table 2 Affinity of compounds to σ ₁ receptor and σ ₂ receptor (inhibition rate %)

Example 61: Acute toxicity study of some compounds

Sequential method limit experiment: ICR mice, half male and half male, were randomly divided into several groups, 2-5 mice in each group, respectively, each compound 2000mg/kg group and solvent group, administered by gavage at 0.2ml/10g . Animals were observed for death within 3 days. (If 3 or more animals survive within three days and there is no obvious abnormality in the life state, continue to observe until the end of the experiment after 7 days. If 3 or more animals die within three days, the median lethal dose method is used Determine its LD50.)

The median lethal dose method pre-test: ICR mice, half male and half male, were randomly divided into several groups, with 4 mice in each group. 10g was administered by gavage, and the death of animals was observed within 1-3 days.

RESULTS: The LD ₅₀ of a single dose of mice was greater than 2000mg/kg, which was comparable to that of the positive control drug S1RA (>2000mg/kg), and had less acute toxicity. The results are shown in Table 3.

Example 62: Partial compound formalin-induced pain model experiment in mice

ICR mice, male, 20-44 g, were randomly divided into negative control group, model group, positive drug dose groups (gabapentin, pregabalin, S1RA) and compound dose groups, with 10 mice in each group. The negative control group and model group were given the corresponding solvent double distilled water by gavage, the positive drug group was given the corresponding positive drug by gavage, and the compound dose groups were given the corresponding dose of compound by gavage, and the gavage volume was 0.1ml/10g. 15 min after gavage, mice were injected subcutaneously with 20 μL of 2.5% formalin in the left hind paw to make a model, and the formation of a skin mound was regarded as the successful standard of modeling. In the negative control group, 20 μL of normal saline was subcutaneously injected into the left hind paw. After successful modeling, observe the time of mice licking and biting the injection foot site at 0-5 minutes and 15-45 minutes after modeling.

Statistical processing of data: mean ± standard deviation (Mean ± SD) of experimental data, one-way analysis of variance was used for comparison; ED ₅₀ was calculated by probability unit regression method. The _ED50 values are shown in Table 3.

Table 3 preferred compound in vivo animal model experimental results

Third Aspect: Composition Examples

Example 63: Tablets

The raw and auxiliary materials are passed through an 80-mesh sieve for use. Weigh the active ingredients, microcrystalline cellulose, lactose, and povidone K30 in the prescribed amount, add them to a high-speed mixing preparation machine, stir and mix at a low speed, add an appropriate amount of purified water, stir at a low speed, and cut at a high speed. Granulate, dry the wet granules at 60°C for 3 hours, granulate with a 24-mesh sieve, add the recipe amounts of sodium starch glycolate, silicon dioxide and magnesium stearate, mix together, and press on a rotary tablet machine.

Those skilled in the art can easily understand that the above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention, etc., All should be included within the protection scope of the present invention.

Claims (9)

1. A phenyl oxadiazole derivative is characterized in that the structural general formula of the phenyl oxadiazole derivative is shown as a formula I or a formula II:

wherein, in the formula I, A is N or CH; m is 0, 1 or 2; z is (CH)₂)_nN is 2,3 or 4;R₁is hydrogen, unsubstituted C_1-5Alkyl or substituted C_1-5An alkyl group; r₂And R₃Each independently selected from fluorine, chlorine, bromine and iodine;

wherein, in the formula I, Z is (CH)₂)_n，R₂And R₃Each independently selected from fluorine, chlorine, bromine and iodine.

2. The phenyloxadiazole derivative of claim 1, wherein said substituted C is_1-5Alkyl being C substituted by fluorine, chlorine, bromine or iodine_1-5An alkyl group.

3. The process for producing a phenyloxadiazole derivative according to claim 1 or 2, which comprises the following reaction formula:

wherein A is N or CH; m is 0, 1 or 2; z is (CH)₂)_nN is 2,3 or 4; r₁Is hydrogen, unsubstituted C_1-5Alkyl or substituted C_1-5An alkyl group; r₂And R₃Each independently selected from fluorine, chlorine, bromine and iodine.

4. A salt of a phenyloxadiazole derivative which is an anion salt of the phenyloxadiazole derivative of claim 1 or 2; the salt of the phenyloxadiazole derivative is hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, methanesulfonate, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate or p-toluenesulfonate.

5. A pharmaceutical composition comprising the phenyloxadiazole derivative of claim 1 or 2 or a salt of the phenyloxadiazole derivative of claim 4.

6. The pharmaceutical composition of claim 5, further comprising an adjuvant or carrier;

preferably, the auxiliary material is a binder, an excipient, a disintegrant, a lubricant or a sweetener;

preferably, the carrier is a grease.

7. Use of a phenyl oxadiazole derivative according to claim 1 or 2 for the manufacture of a medicament for the prevention or treatment of pain; or the use of the phenyloxadiazole derivative salt according to claim 4 for the preparation of a medicament for the prevention or treatment of pain-related diseases.

8. The use according to claim 7, wherein the pain-like disorder is neuropathic pain, cancer pain, angina pectoris, thromboangiitis pain or chest and abdominal pain.

9. The use of claim 7, wherein the medicament is in the form of a tablet, lozenge, capsule, suspension, syrup, paste, lotion or injection.