Pharmaceutical composition for treating diabetic neuralgia and application thereof

Abstract

The invention particularly discloses a pharmaceutical composition for treating diabetic neuralgia and application of the pharmaceutical composition, and relates to the technical field of biological medicines. The invention provides a pharmaceutical composition for treating neuralgia, the pharmaceutical composition comprises infliximab and lipoic acid, and the pharmaceutical composition blocks a TNF-alpha pathway and synergistically resists oxidative stress, so that neuroinflammation and pain are relieved fundamentally, the defects of central side effects and insufficient curative effects of existing drugs are overcome, and a new treatment choice is provided for patients with diabetic neuralgia.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, in particular to a pharmaceutical composition for treating diabetic neuropathic pain and its application. BACKGROUND

[0002] Diabetic neuropathic pain (DNP) is a common chronic complication of diabetes, mainly manifested as tingling, burning or numbness in the distal extremities, and paresthesia. The symptoms of patients usually worsen at night, and with the progression of the disease, infection, foot ulcer, and even amputation can occur, causing great harm to the physical and mental health of patients and seriously affecting the quality of life of patients. With the further increase in the number of global diabetic patients, DNP has become a global health challenge. According to the latest epidemiological data, about half of the diabetic patients develop neuropathy, of which 30%-40% develop neuropathic pain, and about one-fifth of the diabetic patients develop DNP.

[0003] The mechanism of DNP is complex and the specific mechanism has not been clearly elucidated. Current studies show that multiple factors promote the occurrence and development of DNP, which may be related to multiple pathways such as chronic hyperglycemic state, oxidative stress, mitochondrial dysfunction, inflammatory response (such as activation of TNF-α pathway), and increased level of advanced glycation end products.

[0004] Currently, the treatment of DNP mainly involves pain relief on the basis of blood glucose control. The first-line drugs recommended for the treatment of DNP in domestic and foreign guidelines mainly include anticonvulsants such as pregabalin and gabapentin, antidepressants such as amitriptyline, doxepin, and venlafaxine, and opioid analgesics such as tramadol and oxycodone as second-line or third-line drugs. These drugs are usually administered by oral or injection routes and act on the central nervous system to achieve the purpose of relieving pain. However, due to the mechanism of these drugs, patients often experience neurological and psychiatric symptoms such as dizziness, headache, drowsiness, ataxia, and abnormal thinking, liver function damage, gastrointestinal discomfort, and even drug dependence, which affect the quality of life and treatment compliance of patients. Therefore, there is an urgent need to discover new target points to find more effective drugs for treating DNP. SUMMARY

[0005] (I) Technical problems solved In view of this, one of the main purposes of the present application is to provide a pharmaceutical composition for treating diabetic neuropathic pain and its application. By intraperitoneal injection of IFX (infliximab) and lipoic acid, the TNF-α pathway is blocked and the oxidative stress is synergistically inhibited, which fundamentally relieves neuroinflammation and pain, overcomes the defects of central side effects and insufficient efficacy of existing drugs, and provides a new treatment option for diabetic neuropathic pain patients.

[0006] (II) Technical Solution To achieve the above objectives, the present invention provides a pharmaceutical composition for treating neuralgia, wherein infliximab and thioctic acid are the main active substances.

[0007] In one embodiment, the neuralgia is diabetic neuralgia.

[0008] In one embodiment, the dosage ratio of infliximab to thioctic acid is 1:(1~50).

[0009] In one embodiment, the dosage ratio of infliximab to thioctic acid is 1:(10~30).

[0010] In one embodiment, the dosage ratio of infliximab to thioctic acid is 1:20.

[0011] In one embodiment, the dose of infliximab is 2.5 mg / kg and / or the dose of thioctic acid is 50 mg / kg.

[0012] In one embodiment, the infliximab is administered once every two weeks.

[0013] In one embodiment, the thioctic acid is applied once daily.

[0014] In one embodiment, the pharmaceutical composition further includes a pharmaceutically or immunologically acceptable carrier or excipient.

[0015] In another aspect, the present invention provides a pharmaceutical preparation comprising the above-described pharmaceutical composition.

[0016] In one embodiment, the dosage form of the pharmaceutical preparation includes one or a combination of injections, tablets, powders, granules, capsules, pills, liposomes, suppositories, mud patches, or patches.

[0017] In one embodiment, the dosage form of the pharmaceutical preparation is an injection.

[0018] (III) Beneficial Effects This invention provides a pharmaceutical composition for treating diabetic neuropathy and its application. Compared with the prior art, it has the following advantages: 1. For the first time, IFX (anti-inflammatory) and lipoic acid (antioxidant) were combined for DNP via peripheral administration, avoiding central side effects.

[0019] 2. IFX neutralizes TNF-α and inhibits neuroinflammation, while lipoic acid scavenge free radicals to repair oxidative damage. The two work synergistically to downregulate pro-inflammatory factors (TNF-α, IL-6) and upregulate neurotrophic factors (NGF, BDNF).

[0020] 3. Low-dose IFX (2.5 mg / kg) combined with lipoic acid is effective and reduces the risk of immunosuppression. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 This is a graph showing the changes in blood glucose levels in mice with diabetic neuropathy after modeling in Example 2, where **** indicates P<0.0001.

[0023] Figure 2 This is a graph showing the weight change of diabetic neuropathy mice after modeling in Example 2, where **** indicates P<0.0001.

[0024] Figure 3 This is a graph showing the change in mechanical pain threshold after establishing a diabetic neuralgia mouse model in Example 2, where **** indicates P<0.0001.

[0025] Figure 4 This is a graph showing the change in thermal pain threshold after the establishment of a diabetic neuralgia mouse model in Example 2. In the graph, ** indicates P<0.01, and **** indicates P<0.0001.

[0026] Figure 5 The curves showing the changes in mechanical pain threshold in diabetic neuropathy mice after drug administration in Example 4 are shown. In the curves, # indicates P<0.5, ### indicates P<0.001, ☆ indicates P<0.5, and * indicates P<0.5.

[0027] Figure 6 The curves showing the changes in thermal pain threshold in diabetic neuropathy mice after drug administration in Example 4 are shown. In the curves, # indicates P<0.5, ## indicates P<0.01, ### indicates P<0.001, ☆ indicates P<0.5, and * indicates P<0.5.

[0028] Figure 7 The expression of TNF-α, IL-6 (inflammatory factor), NGF, and BDNF (neurotrophic factor) was detected by Western Blot after the grouped administration in Example 4.

[0029] Figure 8 This is a statistical chart showing the normalized analysis of TNF-α, IL-6 (inflammatory factor) and NGF, BDNF (neurotrophic factor) after group administration in Example 4. In the chart, * indicates P<0.5, ** indicates P<0.01, and *** indicates P<0.001. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] Terms and Definitions As used herein, the terms “active substance” or “active substance of the present disclosure” are used interchangeably and refer to infliximab and lipoic acid.

[0032] The active substance disclosed in this invention accounts for 0.001 to 99.9 wt% of the total weight of the composition; preferably 1 to 95 wt% of the total weight of the composition, more preferably 5 to 90 wt%, and more preferably 10 to 80 wt%. The balance consists of pharmaceutically acceptable carriers and other additives.

[0033] As used herein, the term "pharmaceutical composition" refers to a composition comprising infliximab and thioctic acid formulated together with one or more pharmaceutically acceptable carriers.

[0034] The formulation of the pharmaceutical composition can be adjusted according to the application. In particular, the pharmaceutical composition can be formulated using methods known in the art to provide rapid, continuous, or delayed release of the active ingredient upon administration to mammals. For example, the formulation can be selected from any of the following: injections, tablets, powders, granules, capsules, pills, liposomes, suppositories, plasters, or patches.

[0035] As used herein, the term "pharmaceuticalally acceptable" refers to a substance that is suitable for use in humans and / or animals without excessive adverse effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio.

[0036] As used herein, the term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refers to a carrier used for the administration of therapeutic agents, encompassing a variety of excipients and diluents. This term refers to pharmaceutical carriers that are not essential active ingredients themselves and do not cause excessive toxicity upon administration. Suitable carriers are well known to those skilled in the art, and a thorough discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991).

[0037] Pharmaceutically acceptable carriers in a composition include any and all solvents, dispersion media, preservatives, antioxidants, coatings, isotonic and absorption-delaying agents, surfactants, fillers, disintegrants, binders, diluents, lubricants, flow aids, pH adjusters, buffers, enhancers, wetting agents, solubilizers, surfactants, antioxidants, etc., compatible with drug administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The composition may contain other active compounds that provide complementary, additional, or enhanced therapeutic functions. Solid carriers or excipients, such as lactose, starch, or talc, or liquid carriers, such as water, fatty oils, or liquid paraffin, are possible. Other examples of carriers include culture media, such as DMEM or RPMI; and cryogenic storage media containing components that scavenge free radicals, provide pH buffering, osmotic / osmotic support, energy substrates, and ion concentrations to balance intracellular states at low temperatures; and mixtures of organic solvents with water.

[0038] As used herein, the term “treatment” for a symptom or patient refers to steps taken to achieve a beneficial or desired outcome, including clinical outcomes. Beneficial or desired clinical outcomes include, but are not limited to, eliminating, substantially inhibiting, slowing, or reversing the progression of a disease, symptom, or condition; substantially improving or alleviating the clinical or aesthetic symptoms of a symptom; substantially preventing the clinical or aesthetic symptoms of a disease, symptom, or condition; and avoiding harmful or unpleasant symptoms. Treatment also refers to achieving one or more of the following: (a) reducing the severity of the symptom; (b) limiting the development of characteristic symptoms of the symptom being treated; (c) limiting the exacerbation of characteristic symptoms of the symptom being treated; (d) limiting the recurrence of the symptom in patients who previously had the symptom; and / or (e) limiting the recurrence of symptoms in patients who previously did not have symptoms of the symptom.

[0039] As used in this article, “containing,” “having,” or “including” includes “containing,” “mainly composed of,” “substantially composed of,” and “composed of”; “mainly composed of,” “substantially composed of,” and “composed of” are subordinate concepts of “containing,” “having,” or “including.”

[0040] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, and the reagents, methods and equipment used are conventional reagents, methods and equipment in this technical field.

[0041] Example 1: Animal husbandry: Male C57BL / 6J mice aged 8-10 weeks were used. The C57BL / 6J mice were purchased from SPAF Beijing Biotechnology Co., Ltd. The mice were housed in a standard animal husbandry environment and allowed to adapt to the new environment for one week. They had free access to food and water. The housing environment was kept at a constant temperature (23-25℃), constant humidity (40-60%), and a 12-hour light-dark cycle.

[0042] Example 2: Disease Model Establishment Select 7-8 week old male C57 mice, weighing 22-25g. Measure the mice's weight the day before modeling. On the day of modeling, fast the mice but allow them to drink water for at least 4 hours beforehand, and then administer a single intraperitoneal injection of 180mg / kg STZ (Sigma-Aldrich, St. Louis, MO).

[0043] Prior to injection, STZ was rapidly dissolved in freshly prepared sodium citrate buffer (pH 4.5, 50 mM), and control mice were given an equal volume of sodium citrate buffer. Three days after STZ injection, blood samples were collected from the tail vein of mice, and blood glucose levels were measured using a Roche (Switzerland) glucometer. Mice with blood glucose levels ≥16.7 mmol / L were designated as diabetic model mice. Figure 1 , Figure 2 ).

[0044] After establishing a diabetic mouse model, von Frey fibers were used to test the mice's mechanical abnormal pain response to confirm the success of the diabetic neuropathy model. Mice 7 weeks after STZ (180 mg / kg) injection (STZ 7W) exhibited significant spontaneous pain and hyperalgesia, confirming the successful establishment of the diabetic neuropathy model. Figure 3 , Figure 4 ).

[0045] Example 3: Administration: The diabetic neuropathy model mice constructed in Example 2 were randomly divided into 4 groups, with 5 mice in each group: a diabetic neuropathy model control group, an infliximab monotherapy group, a thioctic acid monotherapy group, and a combination therapy group. A normal control group was also included. Mice in the infliximab monotherapy group received intraperitoneal injections of infliximab at a dose of 5 mg / kg every 2 weeks for 4 weeks; mice in the thioctic acid monotherapy group received intraperitoneal injections of thioctic acid at a dose of 100 mg / kg once daily for 4 weeks; mice in the low-dose combination therapy group received intraperitoneal injections of infliximab (2.5 mg / kg) every 2 weeks for 4 weeks and thioctic acid (50 mg / kg / day) once daily for 4 weeks; mice in the model control group and the normal control group received an equal volume of physiological saline intraperitoneally.

[0046] Example 4: Efficacy Verification Pain intensity was assessed using mechanical pain threshold (von Frey) and thermal pain sensitivity (Hargreaves method), measured before administration and weekly after administration.

[0047] Mechanical pain threshold: Von Frey fibers were used to assess abnormal mechanical pain responses in mice. Before testing, mice were placed in transparent plastic boxes on wire mesh for at least 30 minutes each day for three consecutive days to allow them to acclimatize to the testing environment. During testing, von Frey fibers were used to test the withdrawal reflex threshold of the mouse's hind or forefoot. A withdrawal reflex threshold (g) was recorded when the mouse exhibited a withdrawal or paw-licking response as the pressure on the von Frey fibers gradually increased. Tests were performed every 10 minutes, and the average of three tests was calculated to determine the mechanical pain threshold of the mouse.

[0048] Thermal pain sensitivity: Before testing, mice were placed in a transparent plastic chamber on a transparent hot plate apparatus for at least 30 minutes each day for three consecutive days to allow them to acclimatize to the testing environment. During testing, infrared radiation heat sources (IITC, CA, USA) were applied to the mid-sole of the left hind paw of the mouse. The reflex latency (s) was recorded when the mouse exhibited a paw withdrawal or licking response. The thermal laser stimulation of the left hind paw lasted for a maximum of 20 seconds. Testing was performed every 10 minutes, and the average of three tests was calculated to determine the thermal pain threshold of the mouse. After 4 weeks of treatment, the mice were sacrificed, and the sciatic nerve was harvested. The expression of TNF-α, IL-6 (inflammatory factors), NGF, and BDNF (neurotrophic factors) was detected by Western blot.

[0049] Experimental results and analysis: (1) Pain relief: such as Figure 5 and Figure 6As shown, compared with the model control group, both the mechanical pain threshold and thermal pain threshold of the single-drug group and the combination drug group were significantly increased (P<0.05), and the combination drug group was more effective than the single-drug group. In the von Frey test, compared with the model control group mice, the mechanical pain threshold and thermal pain threshold of the mice in the intraperitoneal injection IFX treatment group were reduced, and the analgesic effect lasted for at least 7 days.

[0050] (2) Regulation of inflammation and repair: such as Figure 7 and Figure 8 As shown, after 4 weeks of treatment, the sciatic nerve was harvested. Western blot analysis showed that the expression of TNF-α and IL-6 was significantly lower than that in the model group, while the expression of NGF and BDNF was significantly higher (P<0.05).

[0051] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0052] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A pharmaceutical composition for treating neuralgia, characterized in that, The pharmaceutical composition contains infliximab and thioctic acid.

2. The pharmaceutical composition according to claim 1, characterized in that, The neuralgia mentioned is diabetic neuralgia.

3. The pharmaceutical composition according to claim 1, characterized in that, The dosage ratio of infliximab to thioctic acid is 1:(1~50).

4. The pharmaceutical composition according to claim 3, characterized in that, The dosage ratio of infliximab to thioctic acid is 1:(10~30).

5. The pharmaceutical composition according to claim 4, characterized in that, The dosage ratio of infliximab to thioctic acid is 1:

20.

6. The pharmaceutical composition according to claim 5, characterized in that, The dosage of infliximab is 2.5 mg / kg and / or the dosage of thioctic acid is 50 mg / kg.

7. The pharmaceutical composition according to any one of claims 1-6, characterized in that, The pharmaceutical composition also includes a pharmaceutically or immunologically acceptable carrier or excipient.

8. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation comprises the pharmaceutical composition according to any one of claims 1-7.

9. The pharmaceutical preparation according to claim 8, characterized in that, The dosage forms of the pharmaceutical preparations include one or a combination of injections, tablets, powders, granules, capsules, pills, liposomes, suppositories, mud patches, or plasters.

10. The pharmaceutical preparation according to claim 9, characterized in that, The dosage form of the pharmaceutical preparation is an injection.

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