阅读说明：本技术 Liproxstatin-1在制备治疗神经病理性疼痛制剂中的应用 (Application of Liproxstatin-1 in preparation of preparation for treating neuropathic pain ) 是由 李响 于 2020-05-15 设计创作，主要内容包括：本发明公开了Liproxstatin-1在制备治疗神经病理性疼痛制剂中的应用。发明人通过研究意外发现,Liproxstatin-1可以有效预防、缓解或治疗神经病理性疼痛,特别是物理损伤引起的神经病理性疼痛。通过在术前注射使用Liproxstatin-1,可以有效预防或缓解术后神经病理性疼痛。(The invention discloses an application of Liproxstatin-1 in preparing a preparation for treating neuropathic pain. The inventor unexpectedly finds that the Liproxstatin-1 can effectively prevent, relieve or treat neuropathic pain, particularly neuropathic pain caused by physical injury. By injecting the Liproxstatin-1 before the operation, the neuropathic pain after the operation can be effectively prevented or relieved.)

Application of Liproxstatin-1 in preparation of preparation for preventing, relieving or treating neuropathic pain.

2. Use according to claim 1, characterized in that: the neuropathic pain is neuropathic pain caused by physical injury.

3. Use according to claim 1, characterized in that: the physical damage is surgery induced physical damage.

4. Use according to claim 1, characterized in that: the preparation is in the form of injection.

5. Use according to claim 4, characterized in that: the use method of the injection comprises the following steps:

preoperative injection; and/or

Injections were continued for 3 days post-surgery.

6. Use according to claim 5, characterized in that: the preoperative injection is preoperative 30 min-2 h injection.

7. Use according to claim 4 or 6, characterized in that: the 3-day postoperative continuous injection is once a day injection.

8. Use according to claim 7, characterized in that: the injection time is 8-10 am per day.

9. An agent for preventing, alleviating or treating neuropathic pain, wherein the active ingredient of the agent comprises Liproxstatin-1.

10. The formulation of claim 9, wherein: the preparation is in the form of injection.

Technical Field

The invention relates to a new application of a compound, in particular to an application of Liproxstatin-1 in preparing a preparation for treating neuropathic pain.

Background

Neuropathic pain (Neuropathic pain) is pain excited or caused by primary damage and dysfunction of the nervous system, and the prevalence rate of general population is between 7% and 10%, and the number of affected people in China is more than 1 hundred million according to the conclusion. Besides causing physiological pain of patients, the neuropathic pain also increases the incidence rate of affective disorders such as depression and anxiety, becomes an important cause of psychological diseases, and even a plurality of patients have suicide tendency, so that the quality of life is obviously reduced, which makes the neuropathic pain become a difficult point and a hot point in the field of pain research.

The system of Liproxstatin-1 is named N- [ (3-chlorophenylyl) methyl ] -spiro [ piperidine-4,2' (1' H) -quinoxalin ] -3' -amine, CAS number 950455-15-9, structural formula

Liproxstatin-1 is a potent iron death inhibitor, and the target of action is in the glutathione synthesis pathway, and can inhibit iron death and Gpx 4-/-cell proliferation in a low nanomolar range, and block lipid peroxidation, thereby inhibiting hypoxia inflammatory injury related to iron death. The main therapeutic effects reported in the prior literature are as follows:

lipoxstatin-1 reduces inflammatory factor release by inhibiting iron death, reduces renal lipid peroxidation Injury, and reduces Acute renal Injury caused by rat pancreatitis (Ma D, et al. inhibition of Fe apoptosis A sites acidity kit Injury in Rats with section acid pathways in Dig DisSci.2020Mar 27.doi:10.1007/s 10620-020-.

Lipoxstatin-1 attenuated acute lung injury caused by ischemia-reperfusion of the mouse gut by inhibiting iron Death (LiY, et al. inhibitor of apoptosis-stimulating protein of p53 inhibition of apoptosis and allergy in vitro/reperfusion-induced enzyme lung therapy. cell Death Differ.202Mar 18.doi:10.1038/s 41418-020-0528-x.).

Lipoxstatin-1 relieves myeloglial Cell aggregation and reduces active oxygen concentration by inhibiting iron death, alleviating Morphine-resistant symptoms in rats (Chen X, Lipoxstatin-1 patents Morphine free route toxic Spinal fetroptosis-like Cell death. ACS Chem Neurosci.2019Dec 18; 10(12): 4824) -4833.)

Lipoxstatin-1 protects mouse myocardium from ischemia reperfusion injury by inhibiting iron death, increasing cell survival, decreasing VDAC1levels, and increasing GPX4 expression (Feng Y, et al Lipoxstatin-1 protection of mouse myocardial ischemia reperfusion injury/reperfusion injury by decreasing depletion VDAC1levels and restoring GPX4levels. biochem Biophys Res Commun.2019Dec 10; 520(3): 606) 611.)

Lipoxstatin-1 reduces the expression of tissue reactive oxygen species and infectious cytokines by inhibiting iron death, thereby reducing radiation-induced fibrosis of lung tissue (Li X, et al. Ferroptosis inhibitor infection-induced fibrosis of TGF-. beta.1. J Inflamm (Long). 2019May 29; 16:11.)

Lipoxstatin-1 reduces intestinal Ischemia reperfusion injury by inhibiting iron Death (Li Y, Ischemia-induced ACSL4activation controls to become metastatic-mediated tissue in intestinal Ischemia/reperfusion Difference.2019 Nov; 26(11): 2284. sup. 2299.)

CN109453173A discloses that Liproxstatin-1 treats radiation lung injury diseases. CN111135177A discloses that Liproxstatin-1 can relieve cerebral edema and blood brain barrier permeability caused by cerebral trauma; the Liproxstatin-1 is found to have the effect of improving the dyskinesia, learning and memory disorder caused by brain trauma; proves that the Liproxstatin-1 can obviously improve anxiety and cognitive function caused by TBI; in addition, the protective effect of Liproxstatin-1 is revealed to be related to the reversal of TBI to cause FTH1 and Nrf2 to be up-regulated and NOX2 to be down-regulated.

To date, there is no report that Liproxstatin-1 has an effect of treating neuropathic pain, and it is impossible to infer whether Liproxstatin-1 exerts an effect of relieving neuropathic pain by inhibiting iron death, from the known properties of Liproxstatin-1.

Disclosure of Invention

The invention aims to overcome at least one defect of the prior art and provides application of Liproxstatin-1 in preparing a preparation for treating neuropathic pain.

The technical scheme adopted by the invention is as follows:

in a first aspect of the present invention, there is provided:

application of Liproxstatin-1 in preparation of preparation for preventing, relieving or treating neuropathic pain.

In some examples, the neuropathic pain is neuropathic pain resulting from physical injury.

In some examples, the formulation is in the form of an injection.

In some examples, the injection is used by:

preoperative injection; and/or

Injections were continued for 3 days post-surgery.

In some examples, the preoperative injection is a 30min to 2 hour injection prior to surgery.

In some examples, the 3-day post-operative continuous injection is a once-a-day injection.

The specific injection time can be adjusted according to actual conditions, and in some examples, the injection time is 8-10 am per day.

In some examples, the injection is performed before or after operation, respectively, to achieve better relief of neuropathic pain after operation.

In a second aspect of the present invention, there is provided:

an agent for preventing, alleviating or treating neuropathic pain, wherein the active ingredient of the agent comprises Liproxstatin-1.

In some examples, the formulation is in the form of an injection.

The invention has the beneficial effects that:

the inventor unexpectedly finds that the Liproxstatin-1 can effectively prevent, relieve or treat neuropathic pain, particularly neuropathic pain caused by physical injury. By injecting the Liproxstatin-1 before the operation, the neuropathic pain after the operation can be effectively prevented or relieved.

Drawings

FIG. 1 shows GFAP expression levels in mice of different treatment groups.

Detailed Description

The technical scheme of the invention is further explained by combining experiments.

When neuropathic pain occurs, important information transmission center is in spinal cord, and the inventor finds that the pain symptom can be relieved by using Liproxstatin-1 unexpectedly in the previous research, and the experiment means is used for verification.

Mechanical allodynia determination

The fiber silk with certain bending strength stimulates the sole of the foot, so that the animal can generate leg lifting actions, and when the actions occur to the animal, the fiber silk strength is called as a mechanical pain threshold value. When neuropathic pain occurs, lower force can enable the pain model animal to generate actions such as leg lifting and the like, namely, the mechanical pain threshold value is reduced.

Thermal hyperalgesia assay

Thermal radiation stimulation (noxious stimulation) is generally used, a radiation light source is focused on the soles of the animals, and the time from the beginning of the recording to the shortening of the feet is called thermal stimulation shortening of the feet. When neuropathic pain occurs, the time for thermal stimulation to contract the foot is shortened.

Astrocyte marker assay

Under neuropathic pain stimulation conditions, activated astrocytes release large amounts of cytokines such as nerve growth factor, neurotrophic factor, tumor necrosis factor, IL-1, IL-6, and TNF-a; the release of nerve active substances such as ROS, NO and the like can enable pain afferent neurons at the dorsal horn of the spinal cord to be in a sensitized state and can directly cause pain. GFAP is a specific marker for astrocytes, and its high expression is one of the most common characteristic responses of astrocytes. Activation and proliferation responses of spinal astrocytes and a significant increase in its characteristic marker, GFAP, were observed in various pain models, with down-regulation of GFAP expression indicating pain relief.

By comparing the injection of Liproxstatin-1, measuring the expression level of mechanical hyperalgesia, thermal hyperalgesia and GFAP in spinal cord, it was evaluated whether Liproxstatin-1 can relieve pain.

Experimental Material

Control drugs: dimethyl sulfoxide (DMSO)

Experimental drugs: liproxstatin-1(HY-12726), manufactured by MedChemexpress (Monmouth Junction, NJ, USA) Inc.

Experimental animals: c57BL/6 mice, supplied by the Experimental animals centre, Guangdong province.

Experimental methods

Grouping: all mice were divided into (1) CCI + DMSO groups (i.p. DMSO, an organic solvent, commonly used to solubilize drugs, without pharmacological effect); (2) CCI + LIP group (intraperitoneal injection of Liproxstatin-1); (3) sham groups, 15 mice each.

And (3) treatment: the sciatic nerve ligation pain model was established for both the CCI + DMSO group and the CCI + LIP group, and the preparation method was described in reference to the method proposed by Bennett et al (Gary J Bennett, Xie Y K.A peripheral moneuropate in rat procedure of pain sensory pain in man. pain,1988, 33 (1): 87-107.): the mice were anesthetized, one side of the sciatic nerve was exposed, and the trunk of the sciatic nerve of the mice was lightly ligated with 4 5-0 chromium catgut sutures. Spontaneous pain manifestations of abnormal gait, such as spontaneous foot lifting, toe closing and curling, foot non-landing, paw eversion, lameness and the like, appear 2 days after operation, cold, allodynia, nociceptive thermal stimulation and mechanical stimulation hyperalgesia shown by the model are shown, the hyperalgesia reaches a peak in about 14 days, and can last for more than 2 months. Sham group mice only exposed nerves and were not ligated.

The CCI + DMSO group and the CCI + LIP group are injected with drugs 1 hour before modeling and 3 days after modeling, the CCI + DMSO group is injected with DMSO (10mg/kg) in the abdominal cavity, the CCI + LIP group mice are injected with Liproxstatin-1(10mg/kg) in the abdominal cavity, and the two groups of mice are injected with the same drug volume; sham group mice were not injected with drug.

Three groups of mice respectively carry out mechanical pain sensitivity and thermal pain sensitivity values before modeling and 1, 3, 5, 7 and 10 days after modeling, and the spinal cords of the mice are taken on the 5 th day and the 10 th day to observe the corpuscles with iron death and detect the content of iron under an electron microscope, and GFAP is measured. Mechanorepinephrine was measured by Von Frey cellosilk (purchased from IITC Life Science Inc, usa) stimulation of rat soles; thermal hyperalgesia (purchased from UgoBasile, italy) was measured by heating the soles of rats with a bolometer; GFAP was measured by Western-Blot, wherein GFAP antibody was purchased from CST, USA. Decreased mechanical and thermal hyperalgesia, increased GFAP expression are considered as the onset of pain symptoms, and increased mechanical and thermal hyperalgesia, decreased GFAP expression are considered as the relief of pain symptoms.

The experimental results are as follows:

1) the mechanical pain sensitivity threshold of the mouse is obviously reduced from 1 day after the sciatic nerve ligation until the observation period is finished, and the mechanical pain threshold of the CCI mouse is increased after 1 hour before the operation and three days after the operation of continuous intraperitoneal injection of the Liproxstatin-1 until the observation period is finished.

2) After 1 day after sciatic nerve ligation, the thermal hyperalgesia threshold of the mice is obviously reduced until the end of the observation period, and after 1 hour before operation and three days after continuous intraperitoneal injection of Liproxstatin-1 after operation, the thermal hyperalgesia threshold of the CCI mice is increased until the end of the observation period.

3) The GFAP expression in spinal cord is obviously increased on the 5 th day and the 10 th day after sciatic nerve ligation of mice, and the GFAP expression level can be reduced after 1 hour of preoperative injection and three days after continuous intraperitoneal injection of Liproxstatin-1 after operation.

Relieving effect of Liproxstatin-1 on neuropathic pain

After 1 day after sciatic nerve ligation, the mechanical hyperalgesia threshold of the mice is obviously reduced until the observation period is ended, and after 1 hour before operation and three days after continuous intraperitoneal injection of the Liproxstatin-1 after operation, the CCI mice have the mechanical pain threshold obviously increased at the fifth day after operation compared with the mice without injection of the Liproxstatin-1 until the observation period is ended (Table 1).

TABLE 1 comparison of mechanical pain sensitivity values (X + -SD, g) for postoperative groups of mice

Note: p <0.05 compared to Sham group, # compared to CCI + Veh group, P < 0.05.

Effect of Liproxstatin-1 on neuropathic pain mice thermal hyperalgesia

After 1 day after sciatic nerve ligation, the thermal nociceptive threshold of the mice is obviously reduced until the observation period is ended, and after 1 hour before operation and three days after continuous intraperitoneal injection of the Liproxstatin-1 after operation, the thermal nociceptive threshold of CCI mice is obviously increased on the fifth day after operation compared with that of the mice without injection of the Liproxstatin-1 until the observation period is ended (Table 2).

TABLE 2 comparison of the thermal pain sensitivity values (X + -SD, s) for the groups of mice after surgery

Note: p <0.05 in comparison to Sham group, # in comparison to CCI + Veh group, P <0.05

Inhibition of gliproxstatin-1 on astrocyte aggregation in spinal cord of neuropathic pain mice

Western-Blot shows that GFAP expression in spinal cord is remarkably increased (CCI + Veh) on the 5 th and 10 th days after sciatic nerve ligation of mice, and GFAP expression level (CCI + Lip) can be reduced after 1 hour injection before operation and three days after continuous intraperitoneal injection of Liproxstatin-1 after operation, which is shown in figure 1.