CN111320617A

CN111320617A - Phenyl oxadiazole derivative and preparation method and application thereof

Info

Publication number: CN111320617A
Application number: CN201811544012.5A
Authority: CN
Inventors: 张桂森; 曹旭东; 张译芳; 刘笔锋; 刘欣
Original assignee: Huazhong University of Science and Technology
Current assignee: Huazhong University of Science and Technology
Priority date: 2018-12-17
Filing date: 2018-12-17
Publication date: 2020-06-23

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

Phenyl oxadiazole derivative and preparation method and application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a phenyl oxadiazole derivative and a preparation method and application thereof.

Background

Pain is an unpleasant feeling and emotional feeling, is accompanied by the existing or potential tissue damage, and is a protective reaction mode of the body to the surrounding environment stimulation, but long-term or excessive pain can cause a series of body function changes such as shock and the like, which seriously affect the normal physiological life of people, especially chronic pain can often make patients feel pain and is a cause of diseases, fatalities and disabilities. Neuralgia is a disease which is a serious harm to the life of patients in the pain diseases, and about 8 percent of people in the world are affected by the disease. Neuropathic pain has a sudden, intense and difficult to heal onset, and despite much work done by researchers in the areas of basic and clinical research, there remain significant challenges in clinical research into neuropathic pain and the mechanisms of analgesia. Because the cause and pathogenesis of neuralgia are complicated and profound, no broad-spectrum and specific targeted medicine for neuralgia exists at present. At present, most of the drugs for treating neuralgia clinically are drugs for treating diseases such as depression and epilepsy, which have neuralgia analgesic activity and are found in clinical application. Most of the drugs are used in the treatment of neuralgia, the analgesic effect is often lower than expected, and the treatment response rate is obviously different due to individual difference.

At present, no specific method and effective medicine exist in clinical treatment of neuralgia, neuropsychiatric medicines are generally used for auxiliary treatment clinically, and the medicines mainly comprise 1) neurotransmitter reuptake inhibitors such as antidepressant medicines (such as amitriptyline and duloxetine), which have large side effects and low response rate, and 2) calcium ion channel α₂Delta-ligand antiepileptic drugs (such as gabapentin and pregabalin) with limited effective rate (about 40% and obvious individual difference), 3) opioid receptor analgesics which are susceptible to drug resistance and addiction with obvious adverse reaction, and other new targets (such as NMDA receptor antagonist, α)₂The research on new drugs such as adrenergic receptor agonists and antiarrhythmic drugs has been recently started, but most of them are still in the early stage of research, and a few of them enter first-stage and second-stage clinics. Therefore, the search for a novel neuralgia analgesic drug which is based on a new action mechanism and an action target and has good therapeutic effect on neuralgia is of great significance.

sigma-1 receptor (sigma)₁Receptors) are emerging drug targets in recent years, and particularly show excellent biological activity in neuralgia analgesia. Sigma₁Receptors are distributed primarily in the central nervous system and also widely in peripheral organs. About sigma₁The study of receptor modulation of human pain began in the 90 s, when sigma was found₁The receptor antagonist can enhance the analgesic activity of opioid analgesics₁Agonists would reduce this phenomenon. In acute pain experiments, σ₁The antisense oligodeoxynucleotide of the receptor can obviously enhance the analgesic effect of morphine and opiate mu receptor agonist. Later, more experiments demonstrated σ₁Receptors are effective by acting directly with opioid μ receptors. On the other hand, σ₁The receptor antagonist itself has analgesic effects on pain, particularly neuropathic pain. In the application of sigma₁In formalin-induced pain experiments in mice with receptor gene knockout, there was no significant pain response in phase I and phase II. The same phenomenon occurs in the capsaicin-stimulated induced neuropathic allodynia model. Sigma₁Receptor knockout mice also exhibited pain insensitive behavior in the paclitaxel-induced cold-induced pain hypersensitivity and mechanical stimulus pain hypersensitivity model and the sciatic nerve ligation neuralgia model. Using sigma₁Antagonists of the receptor exhibit sigma-delta in pain model experiments in normal mice₁Receptor knockout mice have a similar phenomenon of desensitization to pain. The experiment proves that the catalyst has sigma₁Haloperidol with receptor antagonism and metabolites I and II thereof have significant analgesic effects in various neuropathic pain models. Sigma₁Similar results were obtained with the receptor antagonists BD-1063 and NE-100. The leading edge in the current study of sigma-1 receptor antagonists is S1RA, which is directed against sigma₁The receptor has high affinity (Ki ═ 17nM) and high selectivity. S1RA is currently used in trials for the treatment of various pains and is entering the clinical stage, where trials for the treatment of neuropathic pain alone have entered clinical stage II.

Therefore, find the selective σ₁The receptor antagonist is used for anti-pain treatment, and has important scientific value and social significance for clinical treatment of pain and neuralgia.

Disclosure of Invention

The invention solves the technical problems of large side effect, low response rate and easy generation of drug resistance and addiction of the pain medicines in the prior art.

According to the first aspect of the present invention, there is provided a phenyl oxadiazole derivative, wherein the structural general formula of the phenyl oxadiazole derivative is represented by 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)_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.

Preferably, said substituted C_1-5Alkyl being C substituted by fluorine, chlorine, bromine or iodine_1-5An alkyl group.

According to another aspect of the present invention, there is provided a preparation method of the phenyloxadiazole derivative, the reaction formula of which 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)_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.

According to another aspect of the present invention, there is provided a salt of a phenyloxadiazole derivative, which is an anion salt of the phenyloxadiazole derivative; the salt of the phenyl oxadiazole 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.

According to another aspect of the present invention there is provided a pharmaceutical composition comprising a phenyl oxadiazole derivative according to claim 1 or 2 or a salt of a phenyl oxadiazole derivative according to claim 3.

Preferably, the pharmaceutical composition further comprises an adjuvant or a carrier;

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

preferably, the carrier is a grease.

According to another aspect of the present invention, there is provided a use of the above-mentioned phenyloxadiazole derivative for the preparation of a medicament for the prevention or treatment of a pain-related disease; or the application of the phenyl oxadiazole derivative salt in preparing a medicament for preventing or treating pain diseases.

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

Preferably, the pharmaceutical is in the form of tablets, lozenges, capsules, suspensions, syrups, ointments, lotions or injections.

Generally, compared with the prior art, the above technical solution conceived by the present invention mainly has the following technical advantages:

(1) the derivative disclosed by the invention is applied to preparation of a medicine 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.

(2) The derivatives and the salt pairs sigma thereof related to the invention₁The receptor has higher affinity with sigma₂Has a low affinity for σ₁The receptor has selective antagonism, which shows that the receptor has analgesic activity. In addition, animal test results also show that the compound can obviously improve phase I and phase II pain induced by formalin. Due to the in vitro action targets, in vivo pharmacological models and sigma₁Receptor-mediated nervous system-mediated responses, particularly pain, are closely related, and therefore the compounds of the present invention have utility in the treatment of pain, particularly neuropathic pain.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

The invention provides a pharmaceutical composition, which comprises a compound shown in formula I or formula II or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable auxiliary materials (such as a carrier and/or an excipient and the like), wherein the pharmaceutical composition contains the compound which can generate pain and neuralgia analgesic activity.

An effective amount of a compound of the present invention may be administered orally, e.g., with an inert diluent or with some carrier. It can be encapsulated in gelatin capsules or compressed into tablets. For the purpose of oral treatment, the compounds of the present invention may be used with excipients and in the form of tablets, troches, capsules, suspensions, syrups and the like. These preparations should contain at least 0.5% by weight of the active compound of the invention, but may vary depending on the particular dosage form, and conveniently comprise from 4% to about 70% by weight of the unit. The amount of active compound in such compositions should be such that a suitable dosage is achieved. Preferred compositions and formulations of the invention contain 1.0 to 300 mg of the active compound of the invention in an oral unit dose.

The compound and the pharmaceutically acceptable salt, solvate and hydrate thereof provided by the invention can be combined with pharmaceutically acceptable carriers or diluents to form a pharmaceutical preparation. 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 depends on the type and severity of the disease or disorder, and also on the characteristics of the subject, such as general health, age, sex, body weight and drug tolerance. The skilled person will be able to determine the appropriate dosage in view of these and other factors. Effective dosages of the cns drug generally employed are well known to the skilled artisan. The total daily dose is usually between about 0.05mg and 2000 mg.

The present invention relates to pharmaceutical compositions which provide from about 0.01 to 1000mg of active ingredient per unit dose. The compositions may be administered by any suitable route, for example orally in the form of capsules, parenterally in the form of injection solutions, topically in the form of ointments or lotions, rectally in the form of suppositories, transdermally in the form of a patch delivery system.

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 and the like contain from about 0.01 to about 99 weight percent of the active ingredient plus a binder such as gelatin, corn starch, gum arabic; excipients such as dibasic calcium phosphate; disintegrating agents such as corn starch, potato starch or alginic acid; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose. When the formulation is in the form of a capsule, it may contain, in addition to the above-mentioned types of raw materials, a liquid carrier such as a fat.

For parenteral administration, the compounds provided herein may be combined with sterile water or an organic medium to form an injectable solution or suspension.

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

In addition, the compounds provided by the invention and pharmaceutical compositions consisting of the compounds can be applied to the treatment and prevention of pain; the pain refers to acute pain, such as acute injury pain of soft tissues and joints, postoperative pain, obstetrical pain, acute herpes zoster pain, gout and the like; chronic pain such as soft tissue and joint strain or degenerative pain, discogenic pain, neurogenic pain, etc.; intractable pain such as trigeminal neuralgia, postherpetic neuralgia, prolapse of intervertebral disc, intractable headache, etc.; cancer pain such as late tumor pain, tumor metastasis pain, and the like; the special pain can be thromboangiitis, intractable angina pectoris, idiopathic chest pain and abdominal pain, etc.

In vitro receptor binding assays indicate that the compounds of the invention are directed to sigma₁The receptor has higher affinity with sigma₂The affinity of (a) is low. To sigma₁Selective antagonism of receptors, tableThe medicine has the potential of analgesic activity.

In addition, animal test results also show that the compound can obviously improve phase I and phase II pain induced by formalin. Due to the in vitro action targets, in vivo pharmacological models and sigma₁Receptor-mediated nervous system-mediated responses, particularly pain, are closely related, and therefore the compounds to which the invention relates have potential in the treatment of pain, particularly neuropathic pain.

In a first aspect: examples of synthetic aspects

Example 1

A process for the preparation of 3- (3, 4-difluorophenyl) -5- (4- (4-methylpiperidin-1-yl) butyl) -1,2, 4-oxadiazole (1) comprising the steps of:

1) adding 80g of 3, 4-difluorobenzonitrile into a 2000mL four-necked bottle, adding 500mL of ethanol, stirring to completely dissolve the 3, 4-difluorobenzonitrile, dissolving 81.2g of hydroxylamine hydrochloride into water, slowly dropwise adding the hydroxylamine hydrochloride into the reaction solution, adding 61g of potassium carbonate powder in batches, heating to 70 ℃, stirring to react, detecting by TLC to complete the reaction, evaporating most of the solvent, adding water, precipitating solids, directly performing suction filtration to obtain white-like solids, adding 70mL of ethyl acetate at room temperature, stirring to dissolve, adding 700mL of petroleum ether, precipitating a large amount of solids, and directly performing suction filtration to obtain a product, wherein the white-like solids are 78g, and the yield is 78.2%.

2) 30g of the product obtained in the first step is taken in a 1000mL single-neck bottle, 400mL of acetone is added and stirred to be dissolved under the condition of ice bath, 26.4g of 5-chlorovaleryl chloride is slowly dropped and stirred to react, and the reaction is detected to be finished by TLC. Directly evaporating most of the solvent, adding saturated sodium bicarbonate aqueous solution, extracting with dichloromethane, taking the organic phase, drying and concentrating to obtain a white-like solid, adding 20mL of ethyl acetate to dissolve the white-like solid, adding 200mL of petroleum ether, stirring at room temperature, separating out a large amount of solid, and directly filtering to obtain 40g of the white-like solid with the yield of 81.1%.

3) And (3) taking 30g of the second-step product, putting the second-step product into a 500mL four-mouth bottle, adding 300mL of toluene, installing a water separation device, heating to 129 ℃, stirring for reacting for 8 hours, and detecting by TLC to finish the reaction. Most of the solvent is directly distilled off, the sample is stirred, the mixture is rinsed by petroleum ether, and column chromatography is carried out to obtain 26g of colorless oily matter, wherein the yield is 95.6%.

4) And taking 2g of the product obtained in the third step, 0.8g of 4-methylpiperidine and 1.7g of potassium carbonate, adding 40ml of acetonitrile, heating to 80 ℃, reacting for 6 hours, cooling to room temperature, directly performing suction filtration, taking mother liquor for concentrating and mixing, leaching with ethyl acetate, and performing column chromatography to obtain 1.0g of colorless oily matter with the yield of 40.7%.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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 shown as follows:

example 2: 3- (3, 4-difluorophenyl) -5- (2- (piperidin-1-yl) ethyl) -1,2, 4-oxadiazole (2)

The title compound was prepared in the same manner as in example 1 except that 3-chloropropionyl chloride was used instead of 5-chlorovaleryl chloride and piperidine was used instead of 4-methylpiperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.90(d,J＝8.8 Hz,1H),7.85(dd,J＝7.1,3.8Hz,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)

The title compound was prepared in the same manner as in example 1 except that 5-chlorovaleryl chloride was changed to 4-chlorobutyryl chloride and 4-methylpiperidine was changed to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The title compound was prepared in the same manner as in example 1 except that 3-chloropropionyl chloride was used instead of 5-chlorovaleryl chloride and morpholine was used instead of 4-methylpiperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.89(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)

The title compound was prepared in the same manner as in example 1 except that 5-chlorovaleryl chloride was changed to 4-chlorobutyryl chloride and 4-methylpiperidine was changed to morpholine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.90(d,J＝9.3 Hz,1H),7.88–7.81(m,1H),7.35–7.26(m,1H),4.33(s,2H),4.04(s,2H), 3.53(s,2H),3.19(dt,J＝14.0,7.1Hz,4H),2.95(s,2H),2.68–2.45(m,2H). 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)

The title compound was prepared by substituting 3-chloropropionyl chloride for 5-chloropentanoyl chloride and 4-methylpiperidine for 4-methylpiperazine for 4-chloropentanoyl chloride in the same manner as in example 1.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.89(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＝6.4 Hz,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 title compound was prepared by substituting 4-chlorobutyryl chloride for 5-chlorovaleryl chloride in the same manner as in example 1.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.90(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＝6.4Hz,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) the title compound was prepared as in example 1 by replacing 4-methylpiperidine with piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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 title compound was prepared as in example 1 by changing 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to 4-methylpiperazine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.89(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 title compound was prepared as in example 1, substituting morpholine for 4-methylpiperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.93–7.88 (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 title compound was prepared in the same manner as in example 1 except that 5-chlorovaleryl chloride was changed to 4-chlorobutyryl chloride and 4-methylpiperidine was changed to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.89(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＝12.8,7.7Hz,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)

The title compound was prepared as in example 1 by substituting 3, 4-difluorobenzonitrile to 2, 4-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride, and 4-methylpiperidine to morpholine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.88(d,J＝8.4 Hz,1H),7.59(d,J＝1.6Hz,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)

The title compound was prepared by the procedure of example 1, substituting 3, 4-difluorobenzonitrile into 2, 4-dichlorobenzonitrile and 5-chlorovaleryl chloride into 4-chlorobutyryl chloride.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.88(d,J＝8.4 Hz,1H),7.58(d,J＝1.4Hz,1H),7.41(d,J＝8.4Hz,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)

The title compound was prepared as in example 1 by substituting 3, 4-difluorobenzonitrile to 2, 4-dichlorobenzonitrile and 4-methylpiperidine to morpholine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.90(d,J＝8.4 Hz,1H),7.57(d,J＝1.7Hz,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)

The title compound was obtained in the same manner as in example 1 by converting 3, 4-difluorobenzonitrile into 2, 4-dichlorobenzonitrile

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.91(d,J＝8.4 Hz,1H),7.58(d,J＝1.6Hz,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)

The objective compound was obtained as in example 1, except that 3, 4-difluorobenzonitrile was changed to 2, 4-dichlorobenzonitrile and 4-methylpiperidine was changed to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.91(d,J＝8.4 Hz,1H),7.58(d,J＝1.8Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 4-dichlorobenzonitrile and 4-methylpiperidine to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.90(d,J＝8.4 Hz,1H),7.56(s,1H),7.39(d,J＝8.4Hz,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)

The title compound was prepared by the procedure of example 1, substituting 3, 4-difluorobenzonitrile into 3, 5-dichlorobenzonitrile and 5-chlorovaleryl chloride into 4-chlorobutyryl chloride.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.96(s,2H),7.52 (s,1H),3.64(d,J＝11.7Hz,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)

The title compound was prepared as in example 1 by substituting 3, 4-difluorobenzonitrile into 3, 5-dichlorobenzonitrile, 5-chlorovaleryl chloride into 4-chlorobutyryl chloride, and 4-methylpiperidine into morpholine

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The title compound was prepared in the same manner as in example 1 by converting 3, 4-difluorobenzonitrile into 3-chloro-4-fluorobenzonitrile and 4-methylpiperidine into morpholine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.15(d,J＝7.0 Hz,1H),8.01–7.93(m,1H),7.28(d,J＝4.0Hz,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)

The objective compound was obtained as in example 1 by converting 3, 4-difluorobenzonitrile into 3-chloro-4-fluorobenzonitrile and converting 4-methylpiperidine into piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.14(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)

The title compound was obtained by converting 3, 4-difluorobenzonitrile into 3-chloro-4-fluorobenzonitrile as in example 1

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.14(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)

The title compound was obtained in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 3-chloro-4-fluorobenzonitrile and 4-methylpiperidine to 4-methylpiperazine

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.14(d,J＝6.3 Hz,1H),7.78(d,J＝8.4Hz,1H),6.85(d,J＝8.4Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 3-chloro-4-fluorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to morpholine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.15(d,J＝7.0 Hz,1H),7.98(dd,J＝7.7,5.3Hz,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)

The objective compound was obtained as in example 1, except that 3, 4-difluorobenzonitrile was changed to 3-chloro-4-fluorobenzonitrile and 5-chlorovaleryl chloride was changed to 4-chlorobutyryl chloride.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The objective compound was obtained as in example 1 by converting 3, 4-difluorobenzonitrile into 3-chloro-4-fluorobenzonitrile and 4-methylpiperidine into pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.15(dd,J＝7.0, 1.8Hz,1H),7.80(d,J＝8.6Hz,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)

The objective compound was obtained as in example 1, except that 3, 4-difluorobenzonitrile was changed to 3-chloro-4-fluorobenzonitrile, 5-chlorovaleryl chloride was changed to 4-chlorobutyryl chloride, and 4-methylpiperidine was changed to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.14(d,J＝6.3 Hz,1H),7.78(d,J＝8.4Hz,1H),6.85(d,J＝8.4Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 3-chloro-4-fluorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to 4-methylpiperazine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The objective compound was obtained as in example 1 by converting 3, 4-difluorobenzonitrile into 2, 5-dichlorobenzonitrile.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.95(d,J＝2.4 Hz,1H),7.50(d,J＝8.6Hz,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)

The title compound was prepared as in example 1 by substituting 3, 4-difluorobenzonitrile to 2, 3-dichlorobenzonitrile and 4-methylpiperidine to morpholine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.78(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.35(t,J＝7.9Hz,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)

The objective compound was obtained as in example 1 by converting 3, 4-difluorobenzonitrile into 2, 3-dichlorobenzonitrile.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.79(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.35(t,J＝7.9Hz,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)

The objective compound was obtained as in example 1, except that 3, 4-difluorobenzonitrile was changed to 2, 5-dichlorobenzonitrile and 4-methylpiperidine was changed to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.94(d,J＝2.0 Hz,1H),7.48(d,J＝8.6Hz,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)

The objective compound was obtained as in example 1, except that 3, 4-difluorobenzonitrile was changed to 2, 3-dichlorobenzonitrile and 4-methylpiperidine was changed to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.79(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.35(t,J＝7.9Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 3-dichlorobenzonitrile and 4-methylpiperidine to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.79(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.36(t,J＝7.9Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 3-dichlorobenzonitrile and 4-methylpiperidine to 4-methylpiperazine.

¹H NMR(400MHz,CDCl₃)δ7.79(d,J＝7.8Hz,1H),7.64(d,J＝8.0Hz, 1H),7.35(t,J＝7.9Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 5-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.92(d,J＝2.4 Hz,1H),7.49(d,J＝8.6Hz,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)

The title compound was prepared by the procedure of example 1, substituting 3, 4-difluorobenzonitrile into 2, 5-dichlorobenzonitrile and 5-chlorovaleryl chloride into 4-chlorobutyryl chloride.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.94(d,J＝2.0 Hz,1H),7.48(d,J＝8.6Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 3-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.00(d,J＝7.9 Hz,1H),7.64(d,J＝8.0Hz,1H),7.36(t,J＝7.9Hz,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) the title compound was prepared in the same manner as in example 1 except for using 2, 3-dichlorobenzonitrile and 4-chlorobutyryl chloride as 3, 4-difluorobenzonitrile and 5-chlorovaleryl chloride, respectively.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.99(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.36(t,J＝7.9Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 3-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride, and 4-methylpiperidine to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.79(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.35(t,J＝7.9Hz,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)

The title compound was prepared as in example 1 by substituting 3, 4-difluorobenzonitrile into 2, 3-dichlorobenzonitrile, 5-chlorovaleryl chloride into 4-chlorobutyryl chloride, and 4-methylpiperidine into morpholine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.79(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.35(t,J＝7.9Hz,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)

The objective compound was obtained as in example 1, except that 3, 4-difluorobenzonitrile was changed to 3, 5-dichlorobenzonitrile and 4-methylpiperidine was changed to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The title compound was prepared in the same manner as in example 1 except that 3, 4-difluorobenzonitrile was changed to 3, 5-dichlorobenzonitrile and 4-methylpiperidine was changed to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The title compound, compound B, was prepared by substituting 3, 4-difluorobenzonitrile into 3, 5-dichlorobenzonitrile and 4-methylpiperidine into morpholine as in example 1,

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The objective compound was obtained as in example 1 by converting 3, 4-difluorobenzonitrile into 3, 5-dichlorobenzonitrile.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 3, 5-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride, and 4-methylpiperidine to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 3, 5-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)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)

The title compound was obtained in the same manner as in example 1 except that 3, 4-difluorobenzonitrile was changed to 2, 4-dichlorobenzonitrile, 5-chlorovaleryl chloride was changed to 4-chlorobutyryl chloride, and 4-methylpiperidine was changed to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.90(d,J＝8.4 Hz,1H),7.57(d,J＝1.7Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 4-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to pyrrolidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.91(d,J＝8.4 Hz,1H),7.58(d,J＝1.6Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 4-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to 4-methylpiperazine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.90(d,J＝8.4 Hz,1H),7.56(s,1H),7.39(d,J＝8.4Hz,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)

The title compound was prepared in the same manner as in example 1 except for changing 3, 4-difluorobenzonitrile to 2, 3-dichlorobenzonitrile, 5-chlorovaleryl chloride to 4-chlorobutyryl chloride and 4-methylpiperidine to 4-methylpiperazine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.79(d,J＝7.8 Hz,1H),7.64(d,J＝8.0Hz,1H),7.35(t,J＝7.9Hz,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)

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.00(d,J＝6.3 Hz,1H),7.91(d,J＝8.4Hz,1H),7.48(d,J＝8.4Hz,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)

¹H NMR(400MHz,CDCl₃)δ8.00(d,J＝6.3Hz,1H),7.89(d,J＝8.4Hz, 1H),7.48(d,J＝8.4Hz,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)

The objective compound was obtained as in example 1, except that 3, 4-difluorobenzonitrile was changed to 3, 4-dichlorobenzonitrile and 4-methylpiperidine was changed to piperidine.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.01(d,J＝6.3Hz, 1H),7.91(d,J＝8.4Hz,1H),7.48(d,J＝8.4Hz,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)

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.95(d,J＝6.3 Hz,1H),7.87(d,J＝8.4Hz,1H),7.48(d,J＝8.4Hz,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)

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ8.00(d,J＝6.3 Hz,1H),7.91(d,J＝8.4Hz,1H),7.48(d,J＝8.4Hz,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)

The title compound was prepared by the procedure of example 1, substituting 3, 4-difluorobenzonitrile into 2, 3-dichlorobenzonitrile and 5-chlorovaleryl chloride into 4-chlorobutyryl chloride.

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.92(d,J＝6.3 Hz,1H),7.90(d,J＝8.3Hz,1H),7.48(d,J＝8.3Hz,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)

The nuclear magnetic data and mass spectral data were:¹H NMR(400MHz,CDCl₃)δ7.89(d,J＝6.4 Hz,1H),7.91(d,J＝8.4Hz,1H),7.48(d,J＝8.4Hz,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]⁺)

TABLE 1 preferred compound numbers and structural formulas prepared in the examples

In a second aspect: examples of the pharmacological aspects

Example 59: sigma₁Preparation of receptor membranes and measurement of ligand affinity (inhibition rate)

σ₁Preparation of acceptor membranes

Cutting off heads of guinea pigs, operating on ice, quickly taking brains, combining tissues into a centrifuge tube, adding 0.01M Tris HCl +0.32M sucrose solution into the centrifuge tube, homogenizing for 4 times in 4 grades of 3-4s, then adding 0.01M Tris HCl +0.32M sucrose solution, adjusting the volume to 10ml/g, adjusting the weight of the homogenized test tube by using a balance, and centrifuging for 10min at 1000 r; adding 0.01M TrisHCl +0.32M sucrose solution into the supernatant, adjusting to 2ml/g, centrifuging at 1000r and 4 deg.C for 10 min; taking the supernatant, centrifuging at 11500r and 4 ℃ for 25 min; adding 0.01M Tris HCl +0.32M sucrose solution into the precipitate to adjust the concentration to 3ml/g, incubating at 25 ℃ for 15min, 11500r, centrifuging at 4 ℃ for 25min, and storing the precipitate at-80 ℃ for later use.

The method comprises the following steps: 1. protein quantification by Bradford method, with reference to 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 assay.

(1) Uniformly dispersing the prepared membrane by using a proper amount of homogenate through a homogenizer, and adding a proper amount of homogenate reference protein to determine a quantitative membrane suspension for later use;

(2) adding 100 mu L of membrane preparation into each reaction tube;

(3) adding 100 mu L B solution into total binding Tube (TB) and 100 mu L haloperidol (10-5M final concentration) into non-specific binding tube (NB);

(4) adding 10 mu L of radioligand [3H ] - (+) -pentazocine into each reaction tube, wherein the final concentration is 32.00, 16.00, 8.00, 4.00, 2.00, 1.00, 0.50 and 0.25nM in sequence;

(5) incubating each reaction tube at 25 ℃ for 3h, after the reaction is finished, rapidly filtering the combined ligand through decompression, fully washing the ligand by using ice-cold test buffer solution, taking out the filter disc, putting the filter disc into a 2ml scintillation cup, adding 1ml of toluene scintillation solution, and uniformly mixing;

(6) and (5) putting the scintillation vial into a liquid scintillation counter for counting.

4、σ₁Competitive receptor binding assays

(1) Firstly, using a proper amount of homogenate to disperse the prepared membrane uniformly by a homogenizer, and adding a proper amount of homogenate to form a 50ml membrane suspension for later use;

(2) adding 100 mu L of membrane preparation into each reaction tube;

(3) 100 μ L B solution was added to the total binding Tube (TB), 100 μ L haloperidol (10-5M final concentration) was added to the non-specific binding tube (NB), and 100 μ L test compound (10-5M final concentration) was added to each test compound specific binding tube (SB);

(4) each reaction tube was filled with 10. mu.L of radioligand [3H ] - (+) -pentazocine (final concentration: 4 nM);

(5) incubating each reaction tube at 25 ℃ for 3h, after the reaction is finished, rapidly filtering the combined ligand through decompression, leading Whatman test paper to be saturated by using 0.25% PEI solution 2h in advance, fully washing the mixed solution by using ice-cold test buffer solution, taking out a filter disc, putting the filter disc into a 2ml scintillation cup, adding 1ml of toluene scintillation solution, and uniformly mixing;

5. Statistical processing of data

TB: summary and constants

NB: non-specific binding constant

SB: binding constant of compound

The inhibition ratio (I%) (TB-SB)/(TB-NB) × 100%

Example 60: sigma₂Preparation of receptor membranes and measurement of ligand affinity (inhibition rate)

σ₂Preparation of acceptor membrane: cutting off heads of guinea pigs, operating on ice, quickly taking brains, combining tissues into a centrifuge tube, adding 0.01M Tris HCl +0.32M sucrose solution into 4 grades of 3-4s for homogenate for 4 times, then adding 0.01M Tris HCl +0.32M sucrose solution, adjusting the volume to 10ml/g, adjusting the weight of the homogenized test tube by using a balance, and centrifuging for 10min at 1000 r; adding 0.01M Tris HCl +0.32M sucrose solution into the supernatant, adjusting to 2ml/g, centrifuging at 1000r and 4 ℃ for 10 min; taking the supernatant, centrifuging at 11000r at 4 ℃ for 30 min; suspending the precipitate with 0.01M Tris HCl +0.32M sucrose solution for 30s, adjusting to 3ml/g, incubating at 25 deg.C for 15min, centrifuging at 11000g for 30min, collecting supernatant, storing at-20 deg.C for more than 12h, and incubating at 50 Mm-Tris.

2. sigma-2 receptor competitive binding assays.

(1) Firstly, uniformly dispersing the prepared membrane by using a proper amount of homogenate (50mM Tris buffer solution, pH 7.4) by using a homogenizer for later use;

(2) adding 100 mu L of membrane preparation and 100 mu L of homogenate into each reaction tube respectively;

(3) add 100. mu.L of homogenate to total binding Tubes (TB), 100. mu.L of 5uM DTG (final concentration 0.5X 10-5M) to non-specific binding tubes (NB), and 100. mu.L of test compound (final concentration 10-5M) to each test compound specific binding tube (SB); 100nM (+) - -NANM screens sigma-1 receptor;

(4) the radioligand 3H-DTG 10. mu.L (final concentration 5nM) was added to each reaction tube (each reaction tube was equipped with 2 parallel tubes, each tube was placed on ice during loading);

(5) incubating each reaction tube at 25 ℃ for 120min, after the reaction is finished, rapidly filtering the combined ligand through decompression, soaking the whatman test paper in 0.5% PEI, fully washing the test paper with ice-cold test buffer solution, taking out the filter disc, putting the filter disc into a 2ml scintillation cup, adding 1ml toluene scintillation solution, and uniformly mixing;

5. Statistical processing of data

TB: summary and constants

NB: non-specific binding constant

SB: binding constant of compound

The inhibition ratio (I%) (TB-SB)/(TB-NB) × 100%

Data statistics processing: the compound is shown in the specification^-5To σ at M concentration₁Receptor and sigma₂The receptor affinity was calculated to obtain the inhibition (%) shown in Table 2.

TABLE 2 Compound vs. sigma₁Receptor and sigma₂Affinity of receptor (% inhibition)

Example 61: acute toxicity study of partial Compounds

Limit experiments by sequential method: ICR mice, each half of male and female, are randomly divided into a plurality of groups, each group comprises 2-5, and each group comprises 2000mg/kg of compound and solvent group, and the administration is performed by intragastric administration according to 0.2ml/10 g. Animals were observed for mortality within 3 days. (if 3 or more than 3 animals survive within three days and the life status is not obviously abnormal, the observation is continued until the experiment is ended after 7 days; if 3 or more than 3 animals die within three days, the LD50 is measured by a median lethality method.)

Half-lethal method pre-test: ICR mice are divided into a plurality of groups of 4 mice each with male and female halves at random, each group comprises 1500mg/kg, 1000mg/kg and 500mg/kg of compounds and solvent groups, the compounds and the solvent groups are administrated by intragastric administration according to 0.2ml/10g, and the death condition of the animals within 1-3 days is observed.

As a result: LD for single drenching of mouse₅₀Greater than 2000mg/kg, and positive control drug S1RA (>2000mg/kg) was comparable with less acute toxicity. The results are shown in Table 3.

Example 62: partial compound formalin-induced mouse pain model experiment

ICR mice, male, 20-44g, were randomly divided into a negative control group, a model group, positive drug dose groups (gabapentin, pregabalin, S1RA) and compound dose groups, 10 per group. And (3) feeding corresponding solvent double distilled water to the negative control group and the model group by intragastric administration, feeding corresponding positive medicine to the positive medicine group by intragastric administration, and feeding corresponding dose of compound to each dose group of compound by intragastric administration, wherein the intragastric volume is 0.1ml/10 g. After the gastric lavage is carried out for 15min, the left hind paw of the mouse is injected with 20 mu L of 2.5 percent formalin for molding subcutaneously, the skin dune is formed as the standard for successful molding, and the left hind paw of the negative control group is injected with 20 mu L normal saline subcutaneously. And observing the time for licking the injection foot part of the mouse at 0-5 min and 15-45 min after the molding is successfully performed.

Data statistics processing: the Mean. + -. standard deviation (Mean. + -. SD) of the experimental data is shown, and the comparison is performed by using a single-factor methodPerforming difference analysis; ED (electronic device)₅₀The calculation is performed by probabilistic regression. ED (electronic device)₅₀The values are shown in Table 3.

TABLE 3 in vivo animal model test results for preferred compounds

In a third aspect: composition examples

Example 63: tablet formulation

Sieving raw materials with a 80-mesh sieve for later use, weighing the active ingredients, microcrystalline cellulose, lactose and polyvidone K30 according to the formula amount, adding into a high-speed mixing preparation machine, stirring and mixing uniformly at low speed, adding a proper amount of purified water, stirring at low speed, cutting and granulating at high speed, drying wet granules for 3h at 60 ℃, granulating with a 24-mesh sieve, adding carboxymethyl starch sodium, silicon dioxide and magnesium stearate according to the formula amount, mixing totally, and tabletting by using a rotary tablet press.

It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.

Claims

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;

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:

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 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.