Novel compounds for the treatment of intractable diseases

Abstract

The present invention relates to a novel compound YJ102 for the prevention or treatment of bladder diseases, a composition for the prevention or treatment of neuropathic pain, neuroinflammation, or inflammatory skin diseases, or for the treatment of intractable neurological diseases including neurogenic bladder. The compound YJ102 of the present invention suppresses neuropathic pain caused by various factors with superior efficacy compared to conventional drugs, improves urinary dysfunction caused by spinal cord injury, restores bladder enlargement and bladder function, suppresses the development of erythema and scaling, which are major symptoms of psoriasis in animal models of psoriasis, suppresses thickening of the epidermal layer, and suppresses splenomegaly caused by psoriasis. Therefore, it can be utilized for the treatment of various intractable neurological diseases or bladder diseases.

Description

Technical Field

[0001] The present invention relates to a composition for preventing or treating neuropathic pain, neuroinflammation, bladder diseases or inflammatory skin diseases of a novel compound YJ102, or a therapeutic use for treating intractable nervous system diseases including neurogenic bladder.

Background Art

[0002] Pain is defined as an unpleasant sensory and emotional experience associated with, or described as, actual or potential tissue damage. It can also refer to pain and sensory disturbances resulting from stimulation of areas in contact with the cerebral cortex and limbic system via neural pathways composed of pain receptors and nerve fibers. It can be said to be a defense mechanism to protect the body and a warning reaction that communicates abnormalities both inside and outside the body. Pain itself is not a disease, so eliminating it does not treat the underlying disease. The causes can be broadly divided into sensory cases due to damage or inflammation of somatic or visceral tissues, and neuropathic cases that occur after nerve damage. Sensory pain includes cutaneous pain, visceral pain, somatic pain, sensory neuralgia, nerve root-referred pain, and somatic-referred pain, while neuropathic pain is caused by dysfunction of the peripheral or central nervous system. Neuropathic pain is pain that arises from damage or dysfunction of the nervous system, and is caused by abnormalities in the transmission or processing of pain signals in the nervous system. Neuropathic pain is caused by nerve compression, constriction, injury, infection, or diseases such as diabetes, multiple sclerosis, or certain medications. The pain is described as a burning or stabbing sensation and is accompanied by numbness or tingling in the affected area. Neuropathic pain is most frequently diagnosed based on symptoms, so any pain typically characterized by burning and / or electric shock pain and / or numbness and / or allodynia is considered neuropathic. Other features of neuropathic pain include hyperalgesia (a very exaggerated sensation of pain in response to stimuli), hyperesthesia (increased sensitivity to normal stimuli), paresthesia (unpleasant abnormal sensations as if there were injury, even though there is no injury), and numbness of the limbs (abnormal sensations such as spontaneous or induced "tingling"). Glial cells, which are non-neuronal cells in the nervous system, are known to be involved in the development and maintenance of neuropathic pain. In particular, microglia and astrocytes in the spinal cord play a crucial role in the pathogenesis of neuropathic pain.Following nerve injury, microglia and astrocytes are activated and secrete pro-inflammatory cytokines and chemokines, which trigger an inflammatory response. This further exacerbates inflammation and damage to nerve fibers. Glial cells also contribute to central sensitization, a phenomenon in which nerve cells in the spinal cord become hypersensitive to pain signals. Therefore, therapies targeting glial cells in the spinal cord are emerging as a potential alternative for treating neuropathic pain. Central sensitization refers to the phenomenon of persistent pain due to changes in the spinal nervous system. Pain stimuli originating in the central nervous system are primarily processed in the spinal dorsal horn.

[0003] Nerve cells constitute the nervous system and are essential for controlling sensory perception, learning, memory, and motor skills. However, once damaged, they not only take a long time to recover to a normal state, but unlike other cells, they are difficult to regenerate, making permanent damage highly likely. In this regard, even among nerve cells, there are differences in the injury response and regenerative capacity depending on the location of the injury. In cases such as spinal cord injury (SCI), which is part of the central nervous system, regeneration failure is more common than in other areas, resulting in not only medical impairments such as motor and urinary tract disorders, but also psychological impairments. The bladder is a hollow, sac-like muscular organ responsible for storing and expelling urine. It is located in the pelvis, behind the pubic symphysis, and is connected to the urethra below and the ureters above. In men, the prostate gland is connected to the lower part of the bladder. The bladder stores approximately 400-500cc of urine and expels the stored urine through the urethra according to nerve signals. When the function of the bladder is damaged, various bladder diseases can occur. The causes of such bladder dysfunction can be broadly divided into two categories. One type of functional impairment is caused by external infection, and the other is caused by physical abnormalities. Of these, functional impairment caused by physical abnormalities can be further distinguished into that caused by abnormalities in the bladder itself and that caused by nerve damage. A representative disease resulting from bladder dysfunction due to nerve damage is overactive bladder syndrome. This refers to a condition in which the bladder becomes hypersensitive due to psychological or pathological factors in the patient, causing the bladder muscles to contract abnormally frequently while urine is being stored in the bladder, resulting in a sudden urge to urinate and frequent urination. Common symptoms of overactive bladder syndrome include frequent urination (8 or more times a day), urinary urgency (inability to control the urge to urinate once it is felt), nocturia (waking up 2 or more times at night to urinate), and urge incontinence (sudden, uncontrollable urge to urinate resulting in urine leakage). Neurogenic bladder, known as a type of overactive bladder syndrome, refers to a disorder characterized by bladder and urethral dysfunction resulting from nerve damage caused by various factors such as diabetes, cerebrovascular disease, brain injury, nerve injury, or spinal cord injury (SCI).Neurogenic bladder can be associated with various complications, primarily urinary tract infections, bladder and kidney injuries, and bladder stones. It is known that when nerve damage causes abnormalities in the central nervous system associated with the urination area, nerve signals leading to the urination area are abnormally transmitted, thus inducing neurogenic bladder. Since such neurogenic bladder is essentially a derivative of a neurological disorder, it is known to recur quickly even after treating overactive bladder syndrome with previously known methods, and the development of treatment methods for it has been sought.

[0004] On the other hand, the human immune system plays a role in protecting the body from external antigens that invade the body, but it has self-tolerance and does not attack its own tissues. However, when the self-tolerance of the immune system is destroyed, and immune cells recognize proteins that are normally expressed by the body's own genes as targets for attack, producing antibodies or triggering T-cell reactions that destroy normal tissue, this is called autoimmunity, and when specific symptoms appear, it is called an autoimmune disease. The inflammatory response, which is an immune response, is a repair mechanism that attempts to regenerate damaged areas caused by chemical substances, bacterial infections, and physical actions that cause matrix changes in the body or tissues. When inflammation occurs due to harmful stimuli, infections, and trauma, vasoactive substances such as inflammatory components are released locally, causing inflammation. If such an inflammatory response persists, it can actually promote mucosal damage and potentially lead to inflammatory diseases such as rheumatoid arthritis, arteriosclerosis, gastritis, and asthma, which are characterized by redness, fever, swelling, pain, and functional impairment. Psoriasis, an inflammatory disease, is a non-infectious, chronic inflammatory skin disease and autoimmune disease that occurs in the skin tissue due to an excessive immune response from abnormally activated T cells. It is one of the intractable immune diseases that must be overcome. When psoriasis develops, it often progresses to a long-term condition and may be accompanied by various other diseases such as psoriatic arthritis, metabolic syndromes, and cardiovascular disease. Psoriasis affects approximately 2-3% of the world's population, and in South Korea, the number of cases increased by approximately 8,500 in the five years since 2012, reaching 168,000, which is equivalent to about 3% of the population. Psoriasis is a disease characterized by the formation of erythematous papules and macules on the skin that are clearly bordered and covered with silvery-white scales of various sizes. Its histological feature is the excessive proliferation of epithelium, and it also presents with a variety of clinical symptoms such as pus, scaling, and rashes. The cause of psoriasis is not yet clearly understood, but possible contributing factors include genetic factors, environmental deterioration or triggering factors, and immunological factors. It is now known that psoriasis is caused by genetic factors, as well as individual lifestyle and environmental factors, which lead to the proliferation of keratinocytes and inflammatory responses. Factors that can worsen or trigger psoriasis include skin trauma, infection, cold and dry climates such as winter, dry skin, stress, and medications. Treatment methods for psoriasis include applying medication to the affected area and phototherapy using ultraviolet light.The aforementioned drugs include corticosteroids, vitamin D derivatives, moisturizers, as well as synthetic vitamin A derivatives, immunosuppressants such as cyclosporine and methotrexate, and steroids. However, long-term application of strong corticosteroids can cause various side effects such as telangiectasia, linear skin atrophy, and skin atrophy. Cyclosporine can cause kidney damage and abnormal blood pressure, and cumulative use of methotrexate can cause liver cirrhosis and fibrosis. [Overview of the project] [Problems that the invention aims to solve]

[0005] The object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of neuropathic pain.

[0006] Furthermore, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of neuroinflammation.

[0007] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of bladder disease.

[0008] Furthermore, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of neurogenic bladder.

[0009] Another object of the present invention is to provide a composition for improving bladder dysfunction.

[0010] Another object of the present invention is to provide a composition for suppressing bladder enlargement.

[0011] Furthermore, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory skin diseases.

[0012] Furthermore, an object of the present invention is to provide a cosmetic composition for the prevention or improvement of inflammatory skin diseases.

[0013] Furthermore, an object of the present invention is to provide applications for the manufacture of pharmaceutical compositions for the prevention or treatment of neuropathic pain, neuroinflammation, bladder diseases, neurogenic bladder, or inflammatory skin diseases.

[0014] Furthermore, an object of the present invention is to provide a use for the manufacture of compositions for improving bladder dysfunction or suppressing bladder enlargement.

[0015] Furthermore, an object of the present invention is to provide a method for treating neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin disease.

[0016] Furthermore, an object of the present invention is to provide a method for treating or improving bladder dysfunction or bladder enlargement. [Means for solving the problem]

[0017] To achieve the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of neuropathic pain comprising a compound of chemical formula 1.

[0018] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of neuroinflammation comprising a compound of chemical formula 1.

[0019] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of bladder disease, comprising a compound of chemical formula 1.

[0020] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of neurogenic bladder, comprising a compound of chemical formula 1.

[0021] Furthermore, the present invention provides a composition for improving bladder dysfunction, comprising a compound of chemical formula 1.

[0022] Furthermore, the present invention provides a composition for suppressing bladder enlargement that contains a compound of chemical formula 1.

[0023] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory skin diseases, comprising a compound of chemical formula 1.

[0024] The present invention also provides a cosmetic composition for preventing or improving inflammatory skin diseases, which contains the compound of Chemical Formula 1.

[0025] The present invention also provides an application for use in the manufacture of a pharmaceutical composition for preventing or treating neuropathic pain, neuroinflammation, bladder diseases, neurogenic bladder or inflammatory skin diseases, which contains the compound of Chemical Formula 1.

[0026] The present invention also provides an application for use in the manufacture of a composition for improving bladder function disorder or suppressing bladder hypertrophy, which contains the compound of Chemical Formula 1.

[0027] The present invention also provides a method for treating neuropathic pain, neuroinflammation, bladder diseases, neurogenic bladder or inflammatory skin diseases.

[0028] Furthermore, the present invention provides a method for treating or improving bladder function disorder or bladder hypertrophy, which contains the compound of Chemical Formula 1.

Advantages of the Invention

[0029] According to the present invention, the compound YJ102 of the present invention suppresses neuropathic pain caused by various causes with an effect superior to that of conventional drugs. In particular, it shows an effect of suppressing neuroinflammation induced by nerve injury, improves dysuria caused by spinal cord injury by suppressing urothelial cell death, breakdown of tight junctions, cell infiltration and bladder inflammation, restores bladder hypertrophy and bladder function, suppresses the occurrence of erythema and scales, which are the main symptoms of psoriasis in a psoriasis animal model, suppresses the thickening of the epidermal layer, and suppresses spleen hypertrophy caused by psoriasis. Therefore, it has the effect of being utilized as an application for preventing or treating neuropathic pain or neuroinflammation, an application for preventing or treating bladder diseases, neurogenic bladder or bladder function disorder, and an application for treating inflammatory skin diseases induced by inflammation such as psoriasis.

Brief Description of the Drawings

[0030] [Figure 1a]This diagram shows the process of creating SCI animal models. [Figure 1b] This diagram shows the process of creating LSS animal models. [Figure 1c] This diagram shows the process of creating SNI animal models. [Figure 1d] This figure shows the results of measuring mechanical allodynia in an SCI animal model after oral administration of YJ102. [Figure 1e] This figure shows the results of measuring mechanical allodynia in an LSS animal model after oral administration of YJ102. [Figure 1f] This figure shows the results of measuring mechanical allodynia in an SNI animal model after oral administration of YJ102. [Figure 1g] This figure shows the results of measuring mechanical allodynia in SCI animal models after intraperitoneal administration of YJ102, gabapentin, or pregabalin. [Figure 1h] This figure shows the results of measuring mechanical allodynia in LSS animal models after intraperitoneal administration of YJ102, gabapentin, or pregabalin. [Figure 1i] This figure shows the results of measuring mechanical allodynia in SNI animal models after intraperitoneal administration of YJ102, gabapentin, or pregabalin. [Figure 2a] This figure shows the results of immunohistochemical analysis of microglial activation in an SNI animal model: Cont: contralateral tissue; Ipsi: ipsilateral tissue. [Figure 2b] This figure shows the results of immunofluorescence chemistry analysis of microglial activation in an SNI animal model: Cont: contralateral tissue; Ipsi: ipsilateral tissue. [Figure 2c] This figure shows the activation of microglia in an SNI animal model, confirmed by Western blot analysis: Cont: contralateral tissue; Ipsi: ipsilateral tissue. [Figure 3a]This figure shows the results of immunohistochemical analysis of astrocyte activation in SNI animal models: Cont: contralateral tissue; Ipsi: ipsilateral tissue. [Figure 3b] This figure shows the activation of astrocytes in an SNI animal model, confirmed by Western blot analysis: Cont: contralateral tissue; Ipsi: ipsilateral tissue. [Figure 4a] This figure shows the expression of inflammatory factors in an SNI animal model, confirmed by RT-PCR analysis. [Figure 4b] This figure shows the expression of inflammatory factors in an SNI animal model, as confirmed by Western blot analysis. [Figure 5a] This figure shows the expression of JAK / STAT3 signaling pathway-related factors, which are part of the inflammation suppression mechanism, analyzed by Western blot in an SNI animal model. [Figure 5b] This figure shows the expression of mTOR signaling pathway-related factors, which are part of the inflammation suppression mechanism, analyzed by Western blot in an SNI animal model. [Figure 5c] This figure shows the expression of TRMP7 in an SNI animal model, analyzed by Western blot. [Figure 5d] This figure shows the expression and activation of JMJD3 in an SNI animal model, analyzed by Western blot. [Figure 6a] This figure shows the effect of YJ102 on improving bladder function in a spinal cord injury model, as confirmed by measuring residual urine volume: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 6b] This figure shows the effect of YJ102 on improving bladder function in a spinal cord injury model, as confirmed by bladder function scores: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 6c] This figure shows the effect of YJ102 on improving bladder function in a spinal cord injury model, as confirmed by the size of the excised bladder: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 6d] This figure shows the effect of YJ102 on improving bladder function in a spinal cord injury model, as confirmed by bladder weight. Sham: Group that underwent only laminectomy and did not inflict injury; Veh: Control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: Group administered YJ102. [Figure 7a] This figure shows bladder enlargement confirmed by H&E staining in a spinal cord injury model: SCI 1d: 1 day after spinal cord injury; Sham: group that underwent only laminectomy and no injury; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 7b] This figure shows the thickness of the urothelium layer, lamina propria, and muscle layer in a spinal cord injury model. [Figure 7c] This figure shows the collagen content in the bladder muscle of a spinal cord injury model, confirmed by Masson's trichrome staining (Blue: collagen): SCI 1d: 1 day after spinal cord injury; Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 7d] This figure shows the collagen / muscle ratio (%) in the bladder in a spinal cord injury model: SCI 1d: 1 day after spinal cord injury; Sham: group that underwent only incisional resection and did not inflict damage; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 8a]This figure shows the expression of ZO-1 and Occludin, tight junction proteins in the bladder urothelium, as confirmed by Western blot analysis in a spinal cord injury model: SCI 1d: Day 1 after spinal cord injury; Sham: Group that underwent only laminectomy and no injury; Veh: Control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: YJ102 administered group. [Figure 8b] This figure shows the results of immunofluorescence staining for occludin (Red) in the urothelial layer in a spinal cord injury model: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 9a] This figure shows the infiltration of MPO (neutrophils) (Green) into the bladder one day after spinal cord injury in a spinal cord injury model, as confirmed by immunofluorescence staining: SCI 1d: 1 day after spinal cord injury; Sham: group that underwent only laminectomy and no injury; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 9b] This graph shows the relative fluorescence intensity of MPO-positive cells in the bladder one day after spinal cord injury in a spinal cord injury model: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 9c] This figure shows the infiltration of ED-1 (macrophage) (Green) into the bladder 7 days after spinal cord injury in a spinal cord injury model, as confirmed by immunofluorescence staining: SCI 7d: 7 days after spinal cord injury; Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 9d]This graph shows the relative fluorescence intensity of ED-1 positive cells in the bladder 7 days after spinal cord injury in a spinal cord injury model: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 9e] This figure shows the results of Western blot analysis of ED-1 in the bladder 7 days after spinal cord injury in a spinal cord injury model: Sham: group that underwent only laminectomy and did not cause injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 10a] This figure shows the TUNEL (Blue) staining results in the urothelial layer 3 days after spinal cord injury in a spinal cord injury model: Sham: group that underwent only laminectomy and did not cause injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 10b] This graph shows the number of TUNEL-positive cells in the urothelial layer 3 days after spinal cord injury in a spinal cord injury model: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 10c] This figure shows the results of Western blot analysis of transection-type caspase-3 (activated caspase-3) in the urothelial layer 3 days after spinal cord injury in a spinal cord injury model: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 10d] This figure shows the results of Western blot analysis of UPIII (urothelial marker) in the urothelial layer 3 days after spinal cord injury in a spinal cord injury model: Sham: group that underwent only laminectomy and did not cause injury; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: group administered YJ102. [Figure 11a]This figure shows the expression of p75 in the urothelial layer on days 1, 3, and 7 after spinal cord injury in a spinal cord injury model, analyzed by RT-PCR: Sham: Group that underwent laminectomy only and did not inflict injury; 1d: Day 1 after spinal cord injury; 3d: Day 3 after spinal cord injury; and 7d: Day 7 after spinal cord injury. [Figure 11b] This figure shows the expression of p75 in the urothelial layer on days 1, 3, and 7 after spinal cord injury in a spinal cord injury model, as confirmed by Western blot analysis: Sham: Group that underwent laminectomy only and did not cause injury; 1d: Day 1 after spinal cord injury; 3d: Day 3 after spinal cord injury; and 7d: Day 7 after spinal cord injury. [Figure 11c] This figure shows the expression of p75 in the urothelial layer one and seven days after spinal cord injury in a spinal cord injury model, as confirmed by immunofluorescence staining: Sham: Group that underwent only laminectomy and did not inflict any further injury; 1d: 1 day after spinal cord injury; and 7d: 7 days after spinal cord injury. [Figure 11d] This figure shows the counting of cells that double-express p75 (Red) / TUNEL (Green) by immunofluorescence staining: Sham: Group that underwent only laminectomy and did not cause further injury; 1d: 1 day after spinal cord injury; 7d: 7 days after spinal cord injury; U: urothelium; L: lamina propria; M: muscle layer; and arrows: double-positive cells for p75 / TUNEL. [Figure 11e] This figure shows the p75-mediated inhibitory effect of YJ102 on urothelial cell death in a spinal cord injury model, as confirmed by p75 RT-PCR analysis: 1d: Day 1 after spinal cord injury; 7d: Day 7 after spinal cord injury; Sham: Group that underwent laminectomy only and did not inflict injury; Veh: Control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: YJ102 administered group. [Figure 11f]This figure shows the p75-mediated inhibitory effect of YJ102 on urothelial cell death in a spinal cord injury model, as confirmed by Western blot analysis: 1d: Day 1 after spinal cord injury; 7d: Day 7 after spinal cord injury; Sham: Group that underwent laminectomy only and did not inflict injury; Veh: Control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ102: YJ102 administered group. [Figure 12a] This figure shows the expression of NGF in macrophages in a spinal cord injury model, as confirmed by RT-PCR analysis: Sham: Group that underwent only laminectomy and did not inflict any further injury; 1d: 1 day after spinal cord injury; 3d: 3 days after spinal cord injury; and 7d: 7 days after spinal cord injury. [Figure 12b] This figure shows the expression of NGF and proNGF in macrophages in a spinal cord injury model, as confirmed by Western blot analysis: Sham: Group that underwent laminectomy only and did not cause injury; 1d: 1 day after spinal cord injury; 3d: 3 days after spinal cord injury; and 7d: 7 days after spinal cord injury. [Figure 12c] This figure shows the expression of NGF in macrophages in a spinal cord injury model, as confirmed by RT-PCR analysis: Sham: Group that underwent only laminectomy and did not inflict any further injury; 1d: 1 day after spinal cord injury; 3d: 3 days after spinal cord injury; and 7d: 7 days after spinal cord injury. [Figure 12d] This figure shows the expression of NGF and proNGF in macrophages in a spinal cord injury model, as confirmed by Western blot analysis: Sham: Group that underwent laminectomy only and did not cause injury; 1d: 1 day after spinal cord injury; 3d: 3 days after spinal cord injury; and 7d: 7 days after spinal cord injury. [Figure 12e] This figure shows the identification of double-positive cells for proNGF (Red) and ED-1 (Green) within macrophages in a spinal cord injury model, as determined by immunofluorescence staining: Sham: group that underwent laminectomy only and did not cause further injury; and SCI(7d): 7 days after spinal cord injury. [Figure 13a]This figure shows the expression of NGF in macrophage strains after inflammation induction (LPS treatment) by RT-PCR: 2h: 2 hours after LPS treatment; 6h: 6 hours after LPS treatment; and 12h: 12 hours after LPS treatment. [Figure 13b] This figure shows the expression of NGF and proNGF in macrophage strains after inflammation induction (LPS treatment) by Western blot analysis: 2h: 2 hours after LPS treatment; 6h: 6 hours after LPS treatment; and 12h: 12 hours after LPS treatment. [Figure 13c] This figure shows the expression of NGF in macrophage strains after inflammation induction (LPS treatment) by RT-PCR: 2h: 2 hours after LPS treatment; 6h: 6 hours after LPS treatment; and 12h: 12 hours after LPS treatment. [Figure 13d] This figure shows the expression of NGF and proNGF in macrophage strains after inflammation induction (LPS treatment) by Western blot analysis: 2h: 2 hours after LPS treatment; 6h: 6 hours after LPS treatment; and 12h: 12 hours after LPS treatment. [Figure 14a] This figure shows the analysis of inflammatory cytokine and chemokine expression in the bladder by RT-PCR in a spinal cord injury model: Sham: group that underwent laminectomy only and did not inflict injury; Veh: control group (administered solution of DMSO, methylpyrrolidinone, and water); and YJ: YJ102 administration group; 1d: 1 day after spinal cord injury; 3d: 3 days after spinal cord injury; and 7d: 7 days after spinal cord injury. [Figure 14b] This figure shows the expression of inflammatory factors in the bladder in a spinal cord injury model, as confirmed by Western blot analysis: Sham: group that underwent only laminectomy and did not inflict injury; Veh: control group (administered a solution of DMSO, methylpyrrolidinone, and water); and YJ: YJ102 administration group; 1d: 1 day after spinal cord injury; 3d: 3 days after spinal cord injury; and 7d: 7 days after spinal cord injury. [Figure 15a] This figure shows the expression of inflammatory factors in macrophage strains after inflammation induction (LPS treatment) using RT-PCR. [Figure 15b]This figure shows a graph quantifying the results of Figure 15a. [Figure 16a] This figure shows a schematic diagram of a study evaluating the efficacy of YJ102 using the IMQ animal model as an animal model for psoriasis. [Figure 16b] This figure shows dorsal images of IMQ animal model mice on Day 5: Sham: control group treated with Vaseline cream; IMQ+Vehicle: psoriasis animal model group created with IMQ; and IMQ+YJ102: psoriasis animal model group administered YJ102. [Figure 16c] This figure shows the PASI scores used to evaluate erythema in animal models of psoriasis: ham: control group treated with Vaseline cream; IMQ+Vehicle: psoriasis animal model group created using IMQ; and IMQ+YJ102: psoriasis animal model group administered YJ102. [Figure 16d] This figure shows the PASI scores used to assess the degree of scaling: ham: control group treated with Vaseline cream; IMQ+Vehicle: psoriasis animal model group created using IMQ; and IMQ+YJ102: psoriasis animal model group administered YJ102. [Figure 17a] This figure shows the H&E staining results of skin tissue in animal models of psoriasis: Sham: control group treated with Vaseline cream; IMQ+Vehicle: psoriasis animal model group prepared using IMQ; and IMQ+YJ102: psoriasis animal model group administered YJ102. [Figure 17b] This figure shows a graph quantifying the thickness of the epidermal layer in animal models of psoriasis: Sham: control group treated with Vaseline cream; IMQ+Vehicle: psoriasis animal model group created using IMQ; and IMQ+YJ102: psoriasis animal model group administered YJ102. [Figure 18a] This figure shows images of spleens extracted from psoriasis animal models on Day 6: Sham: control group treated with Vaseline cream; Vehicle: psoriasis animal model group created using IMQ; and YJ102: psoriasis animal model group administered YJ102. [Figure 18b]This figure shows the weight of the spleen extracted from psoriasis animal models on Day 6: Sham: control group treated with Vaseline cream; Vehicle: psoriasis animal model group prepared using IMQ; and YJ102: psoriasis animal model group administered YJ102. [Best Mode for Carrying Out the Invention]

[0031] The present invention will be described in detail below with reference to the attached drawings and examples of its implementation. However, the following examples are presented as illustrative examples of the present invention, and if it is determined that a specific description of a well-known technology or configuration familiar to those skilled in the art would unnecessarily obscure the gist of the present invention, such detailed description may be omitted, and this will not limit the present invention. The present invention can be modified and applied in various ways within the scope of the claims described below and the equivalents interpreted therefrom.

[0032] Furthermore, the terminology used herein is intended to appropriately describe preferred embodiments of the invention and may vary depending on the intent of the user, operator, or the conventions of the art to which the invention pertains. Therefore, the definitions of these terms should be based on the content throughout this specification. When, throughout the specification, a part of the specification is said to "include" a component, this means that, unless otherwise stated, it may include other components rather than excluding them.

[0033] All technical terms used in this invention are used in the sense that would be generally understood by a person skilled in the art in the relevant field, unless otherwise defined. While preferred methods or samples are described herein, similar or equivalent methods are also included within the scope of this invention. The contents of all publications cited herein as references are incorporated into this invention.

[0034] Throughout this specification, the percentages used to indicate the concentration of a particular substance refer to (w / w)%, (w / v)%, and (v / v)%, respectively, for solid / solid, unless otherwise specified.

[0035] In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of neuropathic pain, comprising a compound of chemical formula 1 (YJ102), its isomer, its solvate, its hydrate, or a salt thereof as an active ingredient.

[0036] [ka]

[0037] In one example, neuropathic pain may be neuropathic pain resulting from nerve damage, or it may be pain resulting from damage or dysfunction of peripheral or central nerves.

[0038] In one embodiment, the nerve injury may be multiple sclerosis, Parkinson's disease, stroke, nerve injury due to diabetes, a tumor or malformation of the spinal cord, spinal cord injury or spinal stenosis induced by trauma or inflammation.

[0039] In one specific example, neuropathic pain includes diabetic peripheral neuropathy, central sensitization, herpes zoster, postherpetic neuralgia, trigeminal neuralgia, complex regional pain syndrome, reflex sympathetic dystrophy, migraine, phantom limb pain, neuropathic pain due to trauma, neuropathic pain due to nerve injury, and spinal stenosis. This may include neuropathic pain due to stenosis, neuropathic pain due to spinal cord injury, neuropathic pain due to chronic diseases (such as multiple sclerosis and HIV), neuropathic pain due to trauma (burning pain), neuropathic pain due to collision (i.e., sciatica, carpal tunnel syndrome, etc.), neuropathic pain due to drug exposure or exposure to harmful chemicals, neuropathic pain due to infection or post-infection, neuropathic pain due to damaged organ function, neuropathic pain due to vascular disease, neuropathic pain due to metabolic disease, neuropathic pain due to cancer or cancer treatment, neuropathic pain due to autoimmune disease, neuropathic low back pain, neuropathic pain due to fibromyalgia, or idiopathic neuropathic pain.

[0040] In one embodiment, the composition of the present invention can suppress the activation of microglia or astrocytes.

[0041] In one embodiment, the composition of the present invention can suppress the expression of JMJD3 (Jumonji domain-containing protein 3), TRPM7 (Transient receptor potential cation channel, subfamily M, member 7), IL-6, iNOS, COX-2, IL-1β, TNF-α, MIP-1α, MIP-1β, MIP-2α, Gro-α, or MCP-1.

[0042] In one embodiment, the composition of the present invention can suppress the phosphorylation of p38MAPK, JAK2, STAT3, S6, or p70S6 kinase.

[0043] In one embodiment, the composition of the present invention can increase the level of H3K27Me3 (Trimethylated histone H3 at lysine 27).

[0044] In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of neuroinflammation comprising a compound of chemical formula 1, its isomer, its solvate, its hydrate, or a salt thereof as an active ingredient. [ka]

[0045] In one embodiment, the neuroinflammation may be due to nerve damage, and the nerve damage may be spinal cord injury or spinal stenosis induced by trauma or inflammation.

[0046] In one specific example, neuroinflammation may be caused by the activation of microglia or astrocytes.

[0047] In one embodiment, the composition of the present invention can suppress the expression of JMJD3 (Jumonji domain-containing protein 3), TRPM7 (Transient receptor potential cation channel, subfamily M, member 7), IL-6, iNOS, COX-2, IL-1β, TNF-α, MIP-1α, MIP-1β, MIP-2α, Gro-α, or MCP-1.

[0048] In one embodiment, the composition of the present invention can suppress the phosphorylation of p38MAPK, JAK2, STAT3, S6, or p70S6 kinase.

[0049] In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of bladder disease, comprising a compound of chemical formula 1 (YJ102), its isomer, its solvate, its hydrate, or its salt as an active ingredient. [ka]

[0050] In one example, the bladder disease may be urinary tract stones, cystitis, prostate cancer, urinary incontinence, hematuria, bladder cancer, urethritis, prostatitis, frequent urination, nocturnal enuresis, urge urination, oliguria, acute cystitis, hypospadias, dysuria, bladder stones, urethral stricture, overactive bladder syndrome, chronic cystitis, urinary tract infection, stress urinary incontinence, chronic prostatitis, delayed urination, or chronic bladder failure.

[0051] In one embodiment, the bladder disorder may be caused by a spinal cord injury, and the spinal cord injury may be caused by trauma or inflammation.

[0052] In one example, the spinal cord injury may be traumatic spinal cord injury, degenerative spinal disease, inflammatory spinal disease, spinal cord tumor, spinal cord malformation, spinal tumor, spinal cord hemorrhage, stroke, spinal cord paralysis due to extramedullary vascular disease, myelitis, multiple sclerosis, amyotrophic lateral sclerosis, or myelopathy.

[0053] In one embodiment, the composition of the present invention can suppress bladder enlargement.

[0054] In one embodiment, the composition of the present invention can increase the expression of ZO-1 and occludin in urothelium.

[0055] In one embodiment, the composition of the present invention can protect urothelial cells and suppress urothelial cell death.

[0056] In one embodiment, the compositions of the present invention can suppress the expression of p75, NGF, proNGF, COX-2, iNOS, IL-6, IL-1β, MIP-1α, MIP-1β, MIP-2α, or Gro-α.

[0057] In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of neurogenic bladder, comprising a compound of chemical formula 1 (YJ102), its isomer, its solvate, its hydrate, or its salt as an active ingredient. [ka]

[0058] In one embodiment, the neurogenic bladder may be caused by a spinal cord injury, and the spinal cord injury may be induced by trauma or inflammation.

[0059] In one example, the spinal cord injury may be traumatic spinal cord injury, degenerative spinal disease, inflammatory spinal disease, spinal cord tumor, spinal cord malformation, spinal tumor, spinal cord hemorrhage, stroke, spinal cord paralysis due to extramedullary vascular disease, myelitis, multiple sclerosis, amyotrophic lateral sclerosis, or myelopathy.

[0060] In one case, neurogenic bladder may present with symptoms of urinary retention, urinary urgency, urge incontinence, frequent urination, nocturia, dysuria, dysuria, or pyelonephritis.

[0061] In one embodiment, the composition of the present invention can suppress bladder enlargement.

[0062] In one embodiment, the composition of the present invention can increase the expression of ZO-1 and occludin in urothelium.

[0063] In one embodiment, the composition of the present invention can protect urothelial cells and suppress urothelial cell death.

[0064] In one embodiment, the compositions of the present invention can suppress the expression of p75, NGF, proNGF, COX-2, iNOS, IL-6, IL-1β, MIP-1α, MIP-1β, MIP-2α, or Gro-α.

[0065] In one embodiment, the composition of the present invention comprises all pharmaceutically acceptable salts of the compound of chemical formula 1, solvates, hydrates, racemates, or stereoisomers that can be prepared therefrom.

[0066] In the present invention, the hydrate is a compound of chemical formula 1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, to which water is bonded by non-covalent intermolecular forces, and may contain stoichiometric or non-stoichiometric amounts of water. Specifically, the hydrate can contain water in a ratio of about 0.25 moles to about 10 moles per mole of active ingredient, and more specifically, it may contain about 0.5 moles, about 1 mole, about 1.5 moles, about 2 moles, about 2.5 moles, about 3 moles, about 5 moles, etc.

[0067] In the present invention, the solvate is a compound of chemical formula 1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, bonded to a non-aqueous solvent by intermolecular forces, and the solvent may be present in stoichiometric or non-stoichiometric amounts. Specifically, the solvate may contain solvent molecules in a ratio of about 0.25 moles to about 10 moles per mole of active ingredient, and more specifically, it may contain about 0.5 moles, about 1 mole, about 1.5 moles, about 2 moles, about 2.5 moles, about 3 moles, about 5 moles, etc.

[0068] The YJ102 compound represented by Chemical Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and as a salt, an acid addition salt formed with a pharmaceutically acceptable free acid is useful. Acid addition salts can be obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrite or phosphorous acid, non-toxic organic acids such as aliphatic mono and dicarboxylic acids, phenyl-substituted alkanoates, hydroxyalkanoates and alkanediates, aromatic acids, and aliphatic and aromatic sulfonic acids. Examples of pharmaceutically non-toxic salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphate chlorides, bromides, iodides, fluorides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malons, succinates, suberates, sebacinates, fumarates, maleates, butin-1,4-dioate, hexane-1,6-dioate, and benzoates. This includes salts, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, terephthalates, benzenesulfonates, toluenesulfonates, chlorobenzenesulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, hydroxybutyrates, glycolates, malates, tartrates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, or mandelates.

[0069] The acid addition salt according to the present invention can be produced by conventional methods, for example, by dissolving the YJ102 compound represented by chemical formula 1 in an excess amount of acidic aqueous solution, and then precipitating the salt using a water-miscible organic solvent, such as methanol, ethanol, acetone, or acetonitrile. Alternatively, it can be produced by evaporating the solvent and excess acid from the mixture to dry it, or by suction filtration of the precipitated salt.

[0070] Furthermore, pharmaceutically acceptable metal salts can be produced using bases. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the insoluble compound salt, and evaporating and drying the filtrate. In this case, sodium, potassium, or calcium salts are preferred as the metal salts from a formulation standpoint. The corresponding salts can also be obtained by reacting an alkali metal salt or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0071] The pharmaceutical composition of the present invention may further contain, in addition to the compound of chemical formula 1, a known therapeutic agent as an active ingredient, and may be used in combination with other known treatments for the treatment of the aforementioned disease.

[0072] In the present invention, the term "prevention" means any action that suppresses or delays the onset, spread, and recurrence of the disease by administering the pharmaceutical composition according to the present invention, and the term "treatment" means any action that improves or favorably alters the symptoms of the disease by administering the composition according to the present invention. A person with ordinary skill in the art to which the present invention belongs will be able to grasp the precise criteria for diseases in which the composition of the present application is effective and determine the degree of improvement, enhancement, and treatment by referring to materials provided by the Korean Medical Association, etc.

[0073] In the present invention, the term "therapeutably effective amount" used in combination with the active ingredient means an amount effective in preventing or treating the aforementioned disease, and the therapeutically effective amount of the composition of the present invention may vary depending on several factors, such as the method of administration, the site of application, and the patient's condition. Therefore, the dosage for use in the human body must be determined appropriately, taking into consideration both safety and efficiency. It is also possible to estimate the amount to be used in humans from the effective amount determined by animal experiments. These considerations when determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; and EW Martin ed., Remington's Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

[0074] The pharmaceutical compositions of the present invention are administered in pharmaceutically effective amounts. In the present invention, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable in medical treatment, without causing side effects. The effective dose level may be determined based on factors including the patient's health status, the type and severity of the disease, the activity of the drug, sensitivity to the drug, method of administration, timing of administration, route of administration and elimination ratio, duration of treatment, and other factors known in the medical field, including drugs that are compounded or used concurrently. The compositions of the present invention may be administered as a single therapeutic agent, in combination with other therapeutic agents, sequentially or concurrently with conventional therapeutic agents, and in single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that provides the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

[0075] The pharmaceutical compositions of the present invention may include carriers, diluents, excipients, or combinations thereof of two or more commonly used in biological formulations. In the present invention, the term "pharmaceutically acceptable" means exhibiting properties that are non-toxic to cells or humans to which the composition is exposed. The carrier is not particularly limited as long as it is suitable for intracellular delivery of the composition, and can be, for example, compounds listed in Merck Index, 13th ed., Merck & Co. Inc., physiological saline, sterile water, Ringer's solution, buffered physiological saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components, and other common additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed. Diluents, dispersants, surfactants, binders, and lubricants may also be added to further formulate the compositions into commonly used dosage forms such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets. Furthermore, the formulations can be suitably developed according to each disease or component using appropriate methods in the relevant art, or methods disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990).

[0076] In one embodiment, the pharmaceutical composition may be one or more dosage forms selected from the group including oral preparations, topical preparations, suppositories, sterile injection solutions, and sprays.

[0077] In the present invention, the term "administration" means providing a given substance to an individual or patient by any suitable method, and can be administered parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically as an injectable dosage form) or orally, depending on the method of administration, and the dosage ranges widely depending on the patient's weight, age, sex, health condition, diet, administration time, method of administration, excretion rate, and severity of pain. Liquid formulations for oral administration of the compositions of the present invention include suspensions, liquid formulations, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to water and liquid paraffin, which are simple diluents commonly used. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories, etc. The pharmaceutical compositions of the present invention can also be administered by any device that allows the active substance to reach the target site. Preferred administration methods and formulations include intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, and intravenous drip injection. Injectable preparations can be manufactured using aqueous solvents such as physiological saline and Ringer's solution, non-aqueous solvents such as vegetable oil, higher fatty acid esters (e.g., ethyl oleate), and alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin), and may contain pharmaceutically acceptable carriers such as stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA), emulsifiers, buffers for pH adjustment, and preservatives to inhibit microbial growth (e.g., phenylmercury nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol).

[0078] In the present invention, the term "individual" means all animals, including humans who have developed or are likely to develop the disease, as well as monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quail, cats, dogs, mice, rats, rabbits, or guinea pigs, and the disease can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to an individual. The pharmaceutical composition of the present invention can be administered in combination with conventional therapeutic agents.

[0079] The pharmaceutical composition of the present invention may further contain pharmaceutically acceptable additives, in which case the pharmaceutically acceptable additives can be starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, corn syrup, acacia gum, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, opa-dry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, and talc. The pharmaceutically acceptable additives according to the present invention are preferably contained in an amount of 0.1 to 90 parts by weight relative to the composition, but are not limited thereto.

[0080] In this invention, the term "expression" generally refers to the cellular process in which a biologically active polypeptide is generated from a DNA sequence and exhibits biological activity within the cell. In this sense, gene expression includes not only the processes of transcription and translation, but also post-transcriptional and post-translational processes that may affect the biological activity of a gene or gene product. These processes include, but are not limited to, RNA synthesis, processing, and transport, as well as polypeptide synthesis, transport, and post-translational modification of polypeptides.

[0081] In the present invention, the expression can be confirmed by measuring the expression level of the gene or mRNA using a nucleic acid sequence, a nucleic acid sequence complementary to the nucleic acid sequence, a primer pair, a probe, or a primer pair and probe that specifically recognizes the nucleic acid sequence and a fragment of the complementary sequence, by polymerase chain reaction, real-time RT-PCR, reverse transcription polymerase chain reaction, competitive polymerase chain reaction, RNase, S1 nuclease assay, in situ hybridization, nucleic acid microarray, Northern blotting, or DNA chip method. Alternatively, the expression can be confirmed using an antibody, antibody fragment, aptamer, avidity multimer, or peptidomimetics that specifically recognizes the full-length protein or a fragment thereof by Western blotting, ELISA (enzyme-linked immunosorbent assay). Protein expression levels can be measured and confirmed using methods such as assay, radioimmunoassay (RIA), radioimmunodiffusion, immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, mass spectrometry, or protein microarrays.

[0082] In one aspect, the present invention relates to a composition for improving bladder dysfunction, comprising a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof as an active ingredient. [ka]

[0083] In one embodiment, the bladder dysfunction may be bladder dysfunction due to spinal cord injury, and the spinal cord injury may be traumatic spinal cord injury, degenerative spinal disease, inflammatory spinal disease, spinal cord tumor, spinal cord malformation, spinal tumor, spinal cord hemorrhage, stroke, spinal cord paralysis due to extramedullary vascular disease, myelitis, multiple sclerosis, amyotrophic lateral sclerosis, or myelopathy.

[0084] In one embodiment, the composition of the present invention can improve or restore the bladder's urination function.

[0085] In one aspect, the present invention relates to a composition for inhibiting bladder enlargement, comprising a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof as an active ingredient. [ka]

[0086] In one embodiment, the composition can suppress the thickening of the bladder wall, i.e., the urothelial layer, lamina propria, or muscular layer of the bladder.

[0087] In one aspect, the present invention relates to a food composition for the prevention or improvement of neuropathic pain, comprising a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof. [ka]

[0088] In one aspect, the present invention relates to a food composition for the prevention or improvement of neuroinflammation comprising a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof. [ka]

[0089] In one aspect, the present invention relates to a food composition for the prevention or improvement of bladder disease comprising a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof. [ka]

[0090] In one aspect, the present invention relates to a food composition for the prevention or improvement of neurogenic bladder comprising the compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof. [ka]

[0091] In one aspect, the present invention relates to a food composition for improving bladder function or suppressing bladder enlargement, comprising a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof.

[0092] When the composition of the present invention is used as a food composition, the compound of chemical formula 1, its isomer, its solvate, its hydrate, or its salt can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The composition may contain food-grade acceptable food additives in addition to the active ingredient, and the amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment).

[0093] The term "food additive" as used in this invention refers to components that can be added to food as an auxiliary, and are added when manufacturing health functional foods in each dosage form, and can be appropriately selected and used by those skilled in the art. Examples of food additives include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and fillers, pectin acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages, but the types of food additives in this invention are not limited by the above examples.

[0094] The food composition of the present invention may include functional foods for health. The term "functional food for health" as used in the present invention refers to a food manufactured and processed into forms such as tablets, capsules, powders, granules, liquids, and pills using raw materials and ingredients that have functional properties useful to the human body. Here, "functional" means obtaining effects useful for health purposes, such as adjusting nutrients or physiological effects, on the structure and function of the human body. The functional foods for health of the present invention can be manufactured by methods commonly used in the ordinary art, and during such manufacturing, raw materials and ingredients commonly added in the ordinary art may be added. Furthermore, the dosage form of the functional food for health can be manufactured without particular limitations, as long as it is a dosage form recognized as a functional food for health. The food composition of the present invention can be manufactured in various dosage forms, and the functional foods for health of the present invention can be taken as an adjunct to enhance the effects of analgesics.

[0095] Furthermore, there are no restrictions on the types of health foods in which the compositions of the present invention can be used. Moreover, compositions containing the compound of chemical formula 1 of the present invention, its isomers, its solvates, its hydrates, or pharmaceutically acceptable salts thereof as active ingredients can be produced by mixing with other appropriate auxiliary ingredients and known additives that can be included in health functional foods, as selected by those skilled in the art. Examples of foods to which it can be added include meats, sausages, bread, chocolates, candies, snacks, confectionery, pizzas, ramen and other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and can be produced by adding it to soups, teas, jellies and juices produced with the compound of the present invention as the main component.

[0096] In one aspect, the present invention relates to the use of the compound of Chemical Formula 1 of the present invention, its isomers, its solvates, its hydrates, or its salts for use in the manufacture of pharmaceutical compositions for the prevention or treatment of neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin diseases.

[0097] In one aspect, the present invention relates to the use of the compound of chemical formula 1 of the present invention, its isomers, its solvates, its hydrates, or its salts for use in the manufacture of compositions for improving bladder dysfunction or suppressing bladder enlargement.

[0098] In one aspect, the present invention relates to a method for treating neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin disease, comprising the step of administering a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof to an individual suffering from neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin disease.

[0099] In one aspect, the present invention relates to a method for treating or improving bladder dysfunction or bladder hypertrophy, comprising the step of administering a compound of chemical formula 1 of the present invention, its isomer, its solvate, its hydrate, or a salt thereof to an individual having symptoms of bladder dysfunction or bladder hypertrophy. [Modes for carrying out the invention]

[0100] The present invention will be described in more detail by the following examples. However, the following examples are provided to illustrate the content of the present invention and do not limit it thereto.

[0101] Example 1. Preparation of an animal model of nerve injury

[0102] 1-1. Creation of animal models of central nervous system injury

[0103] A traumatic spinal cord injury (SCI) model was created for use as an animal model of central nervous system injury (Figure 1a). Specifically, 230-250g male adult Sprague-Dawley rats were anesthetized by intraperitoneal injection of chloral hydrate (500 mg / kg), then the back and nape were shaved to expose the spinal cord without damaging the dura mater, and a laminectomy was performed at the T9-T10 level. To stabilize the spine, the T8 and T11 spinous processes were fixed, and a 10g weight was dropped from a height of 25mm using an NYU impactor onto the dorsal surface of the exposed spinal cord to injure the spine. After that, the muscles and skin were closed, and the rats were left overnight in a temperature and humidity controlled chamber. After surgery, rats were given subcutaneous fluids (5 ml lactated ringer), and antibiotics (gentamicin, 5 mg / kg, intramuscular injection) once daily for 5 days. Body weight, food residue, and water weight were recorded for all animals, and the bladder was emptied twice daily by manual bladder massage until reflex urination was established. The Sham group consisted of rats that underwent laminectomy only and were not injured.

[0104] 1-2. Creation of animal models of peripheral nerve injury

[0105] 1-2-1. Lumbar spinal stenosis model

[0106] A lumbar spinal stenosis (LSS) model was created for use as an animal model of peripheral nerve injury (Figure 1b). Specifically, male adult Sprague-Dawley rats weighing 260-280g were anesthetized by intraperitoneal injection of chloral hydrate (500 mg / kg), then their backs were shaved to expose the vertebral plates at the L4-S2 level. After removing a thin ligamentous flap between L4 and L5, a trapezoidal silicone block (1.00 mm long × 1.3-1.2 mm wide × 1.0 mm high) was inserted into the epidural space beneath the L5 and L6 vertebral plates without damaging the dural sac. In this case, the Sham group was the group in which only a posterior incision was made in the animal, without the insertion of a silicone block.

[0107] 1-2-2. Sciatic nerve injury (SNI) model

[0108] A spinal nerve injury (SNI) model was created for use as an animal model of peripheral nerve injury (Figure 1c). Specifically, male adult Sprague-Dawley rats weighing 200-220g were anesthetized by intraperitoneal injection of chloral hydrate (500 mg / kg). The three peripheral branches of the right sciatic nerve (tibial, common peroneal, and sural nerves) were then approached, and the tibia and peroneal nerves were firmly ligated using 4-0 silk. Nerve resection was performed 3 mm distal to the ligation site. In this study, the sham group consisted of rats in which the sciatic nerve was exposed but not damaged.

[0109] Example 2. Confirmation of the neuropathic pain-suppressing effect of the compound of the present invention.

[0110] To confirm the neuropathic pain-suppressing effect of YJ102 (Chemical Formula 1), a compound of the present invention, rats in which pain was induced 3 weeks (SNI model) or 4 weeks (SCI model and LSS model) post-surgery were selected from the three animal models of nerve injury prepared in Example 1. YJ102 dissolved in a solution of DMSO, methyl pyrrolidinone, and water (1:1.5:1) was administered orally (po) at 10 mg / kg or 20 mg / kg, or intraperitoneally (ip injection) at 1, 2, or 5 mg / kg. The positive control group was administered intraperitoneally at a concentration of 5 mg / kg by dissolving gabapentin or pregavalin in physiological saline, and the vehicle group was administered the same amount of a solution of DMSO, methyl pyrrolidinone, and water. Subsequently, to measure mechanical allodynia, rats were placed at the bottom of a transparent plastic box (10cm x 10cm) on top of a wire mesh (3mm x 3mm) and allowed approximately 20 minutes for adaptation. Von-Frey filaments were applied to the soles of both feet, and tactile (mechanical) sensitivity was assessed using log-increasing rigid nylon Von-Frey single filament series (3.61, 3.84, 4.08, 4.31, 4.56, 4.74, 4.93, and 5.18 mN, Stoelting, WoodDale, IL, USA; equivalent in grams to 0.4, 0.6, 1.0, 2.0, 4.0, 6.0, 8.0, and 15.0). The avoidance response of the feet to increased mechanical stimuli was assessed by applying sufficient force. Von Frey filaments were applied to each foot for 3-4 seconds, to the point where they were fully bent, and the 50% threshold was confirmed using the up-down method (Chaplan et al., 1994). Of the eight Von Frey filaments ranging from 0.4 to 15 g, the medium-weight 2.0 g (4.31 mN) filament was used first to identify the most sensitive area from multiple sites. If no avoidance reaction occurred in the foot, the process was repeated using a thicker filament. If an avoidance reaction occurred, a weaker filament was applied to that sensitive area. The interval between stimulations was 2 seconds.The amount of von Frey hair (grams) that showed a 50% avoidance response to stimulation of the soles of the feet was measured for both feet, and the average value was calculated. Mechanical allodynia was determined to have occurred when the avoidance response threshold was statistically significantly lower compared to before spinal cord injury induction.

[0111] As a result, when mechanical allodynia was measured using von Frey filaments, the PWT (paw withdrawal threshold) score was significantly lower in rats 4 weeks after spinal cord injury, 4 weeks after induction of lumbar spinal stenosis, and 3 weeks after nerve injury compared to normal rats (15 points), showing scores of 2-3 points or less (Figures 1d-f). In all three animal models, the pain threshold was lowered, and it was confirmed that neuropathic pain was induced, where even stimuli that would not normally cause pain were perceived as painful. On the other hand, in all three animal models in which neuropathic pain was induced, oral administration of YJ102 at 10 mg / kg or 20 mg / kg increased the mechanical allodynia threshold starting 30 minutes after administration, and the analgesic effect lasted for more than 4 hours (Figures 1d-f). Furthermore, to compare the effects of YJ102 with those of gabapentin and pregabalin, which are currently prescribed clinically for neuropathic pain, intraperitoneal administration of YJ102 showed that at the same concentration, YJ102 exhibited more than twice the analgesic effect compared to gabapentin or pregabalin, which showed a significant analgesic effect one hour after administration. The duration of the analgesic effect was also more than three times longer than that of the conventional drugs (Figure 1g~i). In particular, while sedation, a drug side effect, was observed when high concentrations of gabapentin (50 mg / kg) and pregabalin (20 mg / kg) were administered, no sedation was observed in the group treated with YJ102, even at a concentration of 100 mg / kg.

[0112] This confirmed that compound YJ102 of the present invention has a significantly greater analgesic effect on neuropathic pain than conventional clinically used drugs (gabapenten, pregabalin), and in particular, it was confirmed that it does not exhibit the side effects observed with conventional drugs, even at high concentrations.

[0113] Example 3. Analysis of the mechanism of suppression of neuropathic pain

[0114] 3-1. Inhibition of Microglia and Astrocyte Activation

[0115] To analyze the activation of spinal microglia, which are known to play a crucial role in the induction and persistence of neuropathic pain, spinal cord tissue from lumbar vertebrae 4-5 was stained with OX-42 four weeks after nerve injury in an SNI animal model, and changes in cell morphology and intensity were observed. Specifically, perfusion was performed through the right atrium of rats in an SNI animal model, fixed with 4% paraformaldehyde, and after transection of spinal nerves (L4-L5), the tissue was embedded in an OCT compound (optimum cutting temperature compound) and sectioned into 10 μm thick sections using a cryostat. Immunohistochemical staining was performed by treating each tissue section with an anti-OX-42 antibody against the OX-42 protein, a marker of microglia, as the primary antibody. After reacting according to the manufacturer's protocol, the tissue was stained with a secondary antibody (Jackson, USA). Furthermore, for protein separation, the severed spinal cord tissue was perfused with PBS, cut into 1 cm sections around the injury site, and homogenized with Lysis Buffer [50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 2 mM MgCl2, 0.1% NP40]. The samples were transferred to microcentrifuge tubes, rocked at 4°C for 20 minutes, and then centrifuged at 25,000 × g for 10 minutes. The supernatant was transferred to a new microcentrifuge, the protein concentration was measured using a BCA kit, and then 1X sample buffer was added and boiled for 5 minutes to prepare the sample for Western blotting. For Western blotting, the same amount of protein was loaded onto an SDS-PAGE and subjected to electrophoresis, then transferred to a nitrocellulose membrane and blotted using primary and secondary antibodies against p38MAPK and p-p38MAPK.

[0116] As a result, microglia stained with OX-42 showed a significant increase in OX-42 intensity and typical activated microglia with thickened and shortened dendrites in the Vehicle-administered group compared to the Sham group, in both the dorsal horn (contralateral) tissue of the control group (non-surgical side) and the ipsilateral tissue of the experimental group (nerve-damaged side) (Figure 2a and b). On the other hand, rats administered with YJ102 showed a significant decrease in these activation changes. Furthermore, analysis of p38MAPK phosphorylation, which is known to be observed only in activated microglia, showed a significant decrease in the amount of p-p38MAPK in rats administered with YJ102 (Figure 2c). This confirmed that YJ102 suppresses microglial activation in spinal cord tissue of a neuropathic pain model.

[0117] 3-2. Inhibition of astrocyte activation

[0118] Using an anti-GFAP antibody against glial fibrillary acidic protein (GFAP), a spinal cord astrocyte activation marker known to play an important role in the induction and persistence of neuropathic pain, the findings were confirmed by immunohistochemical staining and Western blot analysis, similar to Example 3-1.

[0119] As a result, in L4-5 spinal nerve tissue, GFAP expression was significantly increased in the ipsilateral spinal cord compared to the contralateral spinal cord dorsal horn, and astrocyte activation resulting in astrocyte hypertrophy was observed (Figure 3a). On the other hand, in the group administered YJ102, GFAP expression intensity was significantly reduced, which was also shown in the results of Western blot analysis (Figures 3a and b). This confirmed that YJ102 suppresses astrocyte activation in spinal cord tissue of a neuropathic pain model.

[0120] 3-3. Suppression of inflammatory factor expression

[0121] When neuropathic pain occurs, activation of microglia and astrocytes is known to lead to the expression of inflammatory factors, including cytokines and chemokines, causing hyperexcitability of surrounding nerve cells. Therefore, in an SNI animal model 4 weeks after nerve injury, RNA and proteins were extracted from L4-5 spinal cord tissue and analyzed by RT-PCR and Western blot to confirm changes in inflammatory factor expression induced by YJ102 administration. Specifically, for RT-PCR analysis, TRIZOL Reagent (Invitrogen) was used to isolate RNA from spinal cord tissue sections according to the manual. A total mixture of 20 μl was prepared using 1 μg of total RNA, and cDNA was synthesized using MMLV reverse transcriptase (Invitrogen). Using cDNA templates and the primers listed in Table 1 below, the IL-6, iNOS, COX-2, IL-1β, TNF-α, MIP-1α, MIP-1β, MIP-2α, Gro-α, and MCP-1 genes were amplified, and the results were obtained by agarose gel electrophoresis. [Table 1]

[0122] As a result, both RT-PCR and Western blot analysis showed that inflammatory factors whose expression was significantly increased in ipsilateral spinal cord tissue after SNI were significantly decreased in the group treated with YJ102 (Figure 4).

[0123] 3-4. Analysis of signal transduction pathways

[0124] To determine how YJ102 suppressed neuroinflammatory responses, we analyzed changes in factor expression related to the JAK / STAT3 pathway, mTOR pathway, and TRPM7-mediated pathway, which are known to be important regulatory mechanisms and epigenetic regulators mediating neuroinflammatory responses, as well as increased expression and activation of histone demethylase JMJD3, in tissues of an animal model of SNI four weeks after nerve injury.

[0125] As a result, YJ102 significantly reduced the phosphorylation of JAK2 and STAT3, suppressing the JAK / STAT3 pathway and also regulated the mTOR pathway by suppressing the activity of S6 and p70S6 kinases (Figure 5). Furthermore, YJ102 was shown to suppress the expression of TRPM7, a non-selective cation channel reported to cause neuropathic pain by mediating astrocyte activation (Figure 5), confirming that YJ102 is involved in suppressing astrocyte activation by suppressing TRPM7 expression. In addition, YJ102 was shown to significantly suppress the expression of JMJD3, a factor involved in the expression of inflammatory factors such as IL-6, and the activation of H3K27Me3 demethylation through histone demethylation (Figure 5), confirming the possibility that this is how the anti-inflammatory effect occurred.

[0126] Example 4. Confirmation of the bladder function improving effect of the compound of the present invention.

[0127] 4-1. Urinary function analysis

[0128] To confirm the effect of YJ102 (Chemical Formula 1), a compound of the present invention, on urinary dysfunction occurring during spinal cord injury, a traumatic spinal cord injury (SCI) model (Figure 1a) prepared in Example 1 was immediately administered intraperitoneally at a dose of 5 mg / kg after dissolving YJ102 in DMSO, methyl pyrrolidinone, and water (1:1.5:1). This was administered intraperitoneally at 24-hour intervals for 7 days starting the following day. The effect on improving bladder function was then analyzed by residual urine volume testing and the urinary bladder function score (UBFS). In this case, the vehicle group was administered the same amount of DMSO, methyl pyrrolidinone, and water solution. Specifically, to confirm the amount of urine remaining in the bladder after spinal cord injury, bladder expression was performed twice a day by applying pressure to the lowest center of the lower abdomen with the index and middle fingers to empty the bladder. The expelled urine was collected in a urinary collection tube and its volume was recorded. Furthermore, bladder function scores were assessed daily using a 0-4 point UBFS (0 = total loss of function (3 manual bladder pumps per day, blood in urine), 1 = partial recovery of UBF (2 manual pumps per day, blood in urine), 2 = partial recovery of UBF (partial / complete release of sphincter spasms, 1 manual pump per day, persistent blood in urine), 3 = high recovery (1 manual pump per day, no blood in urine), and 4 = normal UBF) in accordance with the reference (Zeng et al., Spinal Cord (2017) 55:834-839).

[0129] The results showed that while normal rats (Sham group) had almost no urine in their bladder because they could urinate on their own, the Vehicle group, which could not urinate on their own after spinal cord injury, showed a significantly increased residual urine volume compared to the Sham group. However, in the YJ102-administered group, residual urine volume decreased significantly from day 1 compared to the Vehicle group (Figure 6a). Furthermore, analysis of bladder function recovery using bladder function scores showed a significant increase in bladder function in the YJ102-administered group compared to the Vehicle group on days 3 and 7 after spinal cord injury (Figure 6b). In addition, bladder size and weight also decreased significantly with YJ102 administration (Figures 6c and 6d).

[0130] 4-2. Analysis of changes in bladder enlargement and bladder muscle collagen content

[0131] As indicators of bladder dysfunction, we examined bladder hypertrophy (thickening of the bladder wall) and changes in collagen content within the bladder muscle. Specifically, we performed perfusion through the left ventricle of an animal model of spinal cord injury, fixed it with 4% paraformaldehyde, removed the bladder, embedded it in an OCT compound (optimum cutting temperature compound), and then sectioned the tissue to a thickness of 10 μm using a cryostat. To measure the thickness of the bladder wall, we performed hematoxylene & eosin (Abcam) staining according to the manufacturer's protocol, and to measure the collagen content within the muscle, we performed Masson's trichrome (Sigma-Aldrich) staining.

[0132] The results showed that the bladder wall thickened significantly on day 1 after spinal cord injury, with the thickness of the urothelium, lamina propria, and muscle layers of the bladder wall increasing 2 to 3 times compared to the normal group. On the other hand, in the group administered YJ102, the thickness of the urothelium, lamina propria, and muscle layers decreased significantly (Figures 7a and b), and the collagen content in the muscle, which had decreased after spinal cord injury, was maintained at almost the same level as the normal group (Figures 7c and d).

[0133] Example 5. Analysis of the treatment mechanism of urinary dysfunction

[0134] 5-1. Suppression of tight junction protein loss in urothelium

[0135] When bladder dysfunction occurs due to reasons such as spinal cord injury or cystitis, a decrease in tight junction proteins in the urothelium, which is in contact with urine, occurs. This increases the permeability of the bladder wall, allowing harmful substances to enter the tissue and leading to urothelial cell death, which plays a barrier role, and an increase in inflammatory responses within the bladder. To confirm how the compound YJ102 of the present invention affects this pathological process, the expression of the tight junction proteins ZO-1 and occludin was confirmed by Western blot analysis and immunofluorescence staining, and the infiltration of neutrophils and macrophages into the bladder was confirmed by immunofluorescence staining. Specifically, the excised bladder tissue sections were frozen in liquid nitrogen, pulverized in a mortar and pestle to completely disrupt them, and homogenized with Lysis Buffer [50mM Tris-HCl (pH 7.4), 150mM NaCl, 2mM MgCl2, 0.1% NP40]. The mixture was transferred to a microcentrifuge tube, rocked at 4°C for 20 minutes, and then centrifuged at 25,000×g for 10 minutes. The supernatant was transferred to a new microcentrifuge, the protein concentration was measured using a BCA kit, then 1X sample buffer was added, and the mixture was boiled for 5 minutes to prepare the sample for Western blotting. For Western blotting, the same amount of protein was loaded onto an SDS-PAGE and electrophoresis was performed. The samples were then transferred to a nitrocellulose membrane and reacted with primary antibodies Occludin (Invitrogen), ZO-1 (Invitrogen), ED-1 (Serotec), and β-actin (Sigma). Finally, blotting was performed using a secondary antibody.Furthermore, for immunofluorescence staining, 10 μm thick bladder tissue sections were treated with ZO-1 (Invitrogen), Occludin (Invitrogen), MPO (DAKO), and ED-1 (Serotec) as primary antibodies, respectively. After reacting according to the manufacturer's protocol, the samples were stained with a secondary antibody (Jackson immunoresults).

[0136] As a result, the levels of ZO-1 and Occludin were significantly reduced on day 1 after spinal cord injury (SCI 1d), and this reduction was significantly suppressed by YJ102 treatment (Figure 8a). Immunofluorescence staining also showed a significant decrease in Occludin expression in the urothelium after spinal cord injury, and this decrease in Occludin was suppressed by YJ102 treatment (Figure 8b). Furthermore, examination of neutrophils and macrophages in bladder tissue confirmed that the infiltration of MPO-positive neutrophils and ED-1-positive macrophages into the bladder after spinal cord injury was significantly suppressed by YJ102 (Figure 9). From this, it was found that YJ102 protected tight junction proteins in the urothelium and blocked the infiltration of immune cells into the bladder.

[0137] 5-2. Suppression of p75-mediated urothelial cell death

[0138] The effect of YJ102 on urothelial cell death in bladder tissue after spinal cord injury was confirmed by TUNEL staining. The protein levels of cleaved caspase-3, which indicates caspase-3 activation, and the protein levels of UPIII, a urothelial marker, were confirmed by Western blot analysis using p75 (Promega), cleaved caspase-3 (Cell Signaling Technology), and UPIII (Abcam) as primary antibodies. Furthermore, to confirm whether the mechanism of cell death is mediated by the p75 neurotrophin receptor, which is thought to be involved in urothelial cell death, the expression level of p75 was confirmed by RT-PCR, Western blot analysis, and immunofluorescence staining. For RT-PCR analysis, the obtained bladder tissue sections were frozen in liquid nitrogen, ground in a mortar, and completely disrupted. RNA was then separated using TRIZOL Reagent (Invitrogen) according to the manual. 20 μl of the total mixture was prepared using 1 μg of total RNA, and cDNA was synthesized using MMLV reverse transcriptase (Invitrogen). p75 gene expression was analyzed using the cDNA template and the primers listed in Table 2 below. [Table 2]

[0139] TUNEL analysis revealed that the number of TUNEL-positive cells in the urothelium was significantly reduced in the YJ102-treated group compared to the vehicle group (Figures 10a and 10b), and caspase-3 activation was also suppressed by YJ102 (Figure 10c). Furthermore, after spinal cord injury, the level of UPIII, a urothelial marker, was significantly reduced (signifying urothelial loss), and this was shown to be suppressed by YJ102 administration (Figure 10d). In addition, after spinal cord injury, the expression of the p75 gene (mRNA) and protein was significantly increased (Figures 11a-c), and it was confirmed that urothelium expressing p75 was TUNEL-positive, confirming that p75 mediates the process of urothelial cell death (Figure 11d). This increased p75 expression was shown to be significantly reduced by YJ102 treatment (Figures 11e and 11f).

[0140] This confirmed that YJ102 has a cytoprotective effect that suppresses urothelial cell death, and that this effect is mediated by p75 receptor signaling.

[0141] 5-3. Suppression of proNGF expression

[0142] To determine whether YJ102 affects the production of proNGF, a ligand for p75, proNGF expression in the bladder was confirmed by RT-PCR, Western blotting, and immunofluorescence staining. For further verification, the effect of YJ102 on proNGF production was investigated by stimulating the macrophage strain Raw 264.7 cell line (Korea Cell Cline Bank, Seoul, Korea) with 1 μg / ml of LPS. YJ102 was dissolved in DMSO and administered 30 minutes prior to LPS treatment.

[0143] As a result, YJ102 significantly suppressed NGF mRNA expression and proNGF production in bladder tissue after spinal cord injury (Figure 12a-d), and proNGF was confirmed to be expressed in macrophages (ED-1 positive cells) that infiltrated the bladder (Figure 12e). When this was confirmed in macrophage strains, it was shown that YJ102 significantly suppressed the expression of proNGF mRNA and protein (Figure 13).

[0144] This means that YJ102 has a cytoprotective effect that suppresses the expression of p75 and its ligand proNGF in the urothelium of the bladder after spinal cord injury, thereby inhibiting urothelial cell death.

[0145] 5-4. Suppression of inflammatory factors

[0146] To confirm the effect of YJ102 on the expression of inflammatory factors in the bladder after spinal cord injury, the expression of inflammatory cytokines and chemokines COX-2, iNOS, IL-6, IL-1β, MIP-1α, MIP-1β, MIP-2α, and Gro-α in bladder tissue was confirmed by RT-PCR analysis and Western blot analysis using the primers shown in Table 2 above. RT-PCR analysis was also performed in the Raw 264.7 cell line.

[0147] As a result, it was shown that YJ102 administration significantly reduced the expression of COX-2, iNOS, IL-6, IL-1β, MIP-1α, MIP-1β, MIP-2α, and Gro-α (Figure 14). In macrophage strains, it was confirmed that the mRNA levels of IL-1β, TNF-α, IL-6, iNOS, and COX-2, which were increased by LPS treatment, significantly decreased with YJ102 treatment (Figure 15).

[0148] This confirmed that, after traumatic spinal cord injury, YJ102 can restore bladder function by reducing urothelial cell death, tight junction breakdown, cell infiltration, and the associated inflammatory response in the bladder, thereby suppressing deformation of bladder tissue.

[0149] Example 6. Preparation of an animal model of psoriasis

[0150] Psoriasis, caused by the overproduction of skin cells, is a chronic skin disease associated with various factors, including autoimmune diseases. Its symptoms include redness, thickening of the skin, and abnormal scaling. To create an animal model of psoriasis that exhibits key indicators of the disease similar to those seen in clinical practice, such as epidermal thickening, immune cell infiltration into the dermis and epidermis, abnormal keratosis, inflammatory response, and activation of the IL-23 / IL-17 cytokine axis, imiquimod (IMQ) was applied to mice. Specifically, the backs of C57BL / 6 mice (male, 18-20g, 8 weeks old) (Daehan Biolink, Eumseong, Korea) were shaved to remove hair, and then IMQ cream (83mg / day) (5%) was applied to the backs for five consecutive days from Day 0 to Day 4 to induce skin inflammation similar to that of psoriasis. Normal control mice (Sham) were treated with Vaseline cream (Figure 16a).

[0151] Example 7. Confirmation of the therapeutic effect of the compound of the present invention on psoriasis.

[0152] 7-1. Relief of erythema and scaling

[0153] To confirm the efficacy of YJ102 (Chemical Formula 1), the compound of the present invention, against psoriasis, YJ102 dissolved in DMSO, methyl pyrrolidinone, and water (1:1.5:1) was administered intraperitoneally (5 mg / kg) for a total of three days on Day 2, Day 3, and Day 4. The Vehicle group received the same amount of solution in DMSO, methyl pyrrolidinone, and water (1:1.5:1) (Figure 16a). Skin changes were assessed daily using the PASI score for erythema and scaling. The PASI score is assessed on a severity scale from 0 to 4, where 0 is none, 1 is mild, 2 is moderate, 3 is severe, and 4 is very severe (PASI reference: Yashpal Manchanda, Abhishek De, Sudip Das, and Disha Chakraborty. Disease Assessment in Psoriasis. Indian J Dermatol. 2023 May~Jun; 68(3):278~281.).

[0154] As a result, in the psoriasis mouse model prepared in Example 1 (Vehicle group), erythema and scaling began to appear on the skin from Day 2 after application of IMQ, and the symptoms worsened on Day 4 and Day 5, with erythema and scaling observed across the entire back. On the other hand, in the group administered YJ102, the degree of erythema and scaling was significantly lower compared to the Vehicle group, and it was confirmed that the symptoms were alleviated by Day 6 (Figures 16b-d).

[0155] 7-2. Suppression of Epidermal Thickening

[0156] On Day 6, skin tissue from a psoriasis mouse model was stained with hematoxyline and eosin, and the thickness of the epidermal layer was measured. Specifically, skin was excised from the back of the mice on Day 6, fixed by immersion in 4% paraformaldehyde for 24 hours, and then paraffin blocks were prepared. 5 μm thick paraffin tissue sections were then prepared. To measure the thickness of the epidermal layer, hematoxyline and eosin staining of the paraffin tissue was performed according to the manufacturer's protocol. Five locations on the epidermal layer of each mouse were photographed under a 200x optical microscope, and the thickness of the epidermal layer was measured from a total of 5 mice (25 sections in total), and the average value was calculated.

[0157] The results showed that the epidermal thickness of the normal control group (Sham group) was 36.6 ± 2.3 μm, while the epidermal thickness of the psoriasis-induced group (Vehicle group) was 141.3 ± 7.5 μm, approximately 38 times thicker than that of the Sham group. On the other hand, the epidermal thickness of the group administered YJ102 was 89.8 ± 6.8 μm, a reduction of approximately 35% compared to the Vehicle group. Furthermore, while a very large number of cells infiltrating the dermis were observed in the skin tissue of the Vehicle group, this number was reduced to the Sham level in the YJ102-administered group (Figure 17).

[0158] This confirmed that YJ102 can alleviate the symptoms of psoriasis.

[0159] Example 8. Confirmation of the inhibitory effect of the compound of the present invention on spleen enlargement caused by psoriasis.

[0160] In mice in which psoriasis was induced, a phenomenon of spleen enlargement occurred, and the longitudinal diameter of the spleen in psoriasis patients increased compared to non-psoriasis patients. Since a correlation between disease duration and spleen diameter has been reported, the spleens of each group of mice were removed on Day 6, and their weight was measured to confirm the degree of spleen enlargement.

[0161] As a result, spleen size and weight were significantly reduced in the YJ102 group compared to the vehicle group (Figure 18). Therefore, the finding that YJ102 reduced splenic hypertrophy in the IMQ psoriasis animal model suggests that YJ102 may modulate immune system abnormalities in psoriasis patients.

Claims

1. A pharmaceutical composition for the prevention or treatment of neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin disease, comprising a compound of chemical formula 1, its isomer, its solvate, its hydrate, or its salt as an active ingredient. 【Chemistry 1】

2. The pharmaceutical composition according to claim 1, wherein neuropathic pain includes pain caused by damage or dysfunction of peripheral or central nerves.

3. Neuropathic pain includes diabetic peripheral neuropathy, central sensitization, herpes zoster, postherpetic neuralgia, trigeminal neuralgia, complex regional pain syndrome, reflex sympathetic dystrophy, migraine, phantom limb pain, neuropathic pain due to trauma, neuropathic pain due to nerve injury, and spinal stenosis. The pharmaceutical composition according to claim 1, wherein the neuropathic pain is due to stenosis, spinal cord injury, chronic disease (multiple sclerosis, HIV, etc.), trauma (burning pain), collision (i.e., sciatica, carpal tunnel syndrome, etc.), drug exposure or exposure to harmful chemicals, infection or post-infection neuropathic pain, damaged organ function, vascular disease, metabolic disease, cancer or cancer treatment, autoimmune disease, neuropathic low back pain, fibromyalgia, or idiopathic neuropathic pain.

4. The pharmaceutical composition according to claim 1, which inhibits the activation of microglia or astrocytes.

5. The pharmaceutical composition according to claim 1, which suppresses the expression of p75, NGF, proNGF, IL-6, JMJD3 (Jumonji domain-containing protein 3), TRPM7 (Transient receptor potential caution channel, subfamily M, member 7), IL-6, iNOS, COX-2, IL-1β, TNF-α, MIP-1α, MIP-1β, MIP-2α, Gro-α, or MCP-1.

6. The pharmaceutical composition according to claim 1, which inhibits the phosphorylation of p38MAPK, JAK2, STAT3, S6, or p70S6 kinase.

7. The pharmaceutical composition according to claim 1, which increases H3K27Me3 (Trimethylated histone H3 at lysine 27).

8. The pharmaceutical composition according to claim 1, wherein the neuroinflammation is induced by the activation of microglia or astrocytes.

9. The pharmaceutical composition according to claim 1, wherein the bladder disease is urinary tract stones, cystitis, prostate cancer, urinary incontinence, hematuria, bladder cancer, urethritis, prostatitis, frequent urination, nocturnal enuresis, urge urination, oliguria, acute cystitis, hypospadias, dysuria, bladder stones, urethral stricture, overactive bladder syndrome, chronic cystitis, urinary tract infection, stress urinary incontinence, chronic prostatitis, delayed voiding, or chronic bladder failure.

10. The pharmaceutical composition according to claim 9, wherein the bladder disorder is caused by spinal cord injury.

11. The pharmaceutical composition according to claim 10, wherein the spinal cord injury is traumatic spinal cord injury, degenerative spinal disease, inflammatory spinal disease, spinal cord tumor, spinal cord malformation, spinal tumor, spinal cord hemorrhage, stroke, spinal cord paralysis due to extramedullary vascular disease, myelitis, multiple sclerosis, amyotrophic lateral sclerosis, or spinal stenosis.

12. The pharmaceutical composition according to claim 1, wherein the neurogenic bladder is caused by spinal cord injury.

13. The pharmaceutical composition according to claim 12, wherein the spinal cord injury is traumatic spinal cord injury, degenerative spinal disease, inflammatory spinal disease, spinal cord tumor, spinal cord malformation, spinal tumor, spinal cord hemorrhage, stroke, spinal cord paralysis due to extramedullary vascular disease, myelitis, multiple sclerosis, amyotrophic lateral sclerosis, or spinal stenosis.

14. The pharmaceutical composition according to claim 1, wherein neurogenic bladder exhibits symptoms of urinary retention, urinary urgency, urge incontinence, frequent urination, nocturia, difficulty urinating, urinary dysfunction, or pyelonephritis.

15. The pharmaceutical composition according to claim 1, which alleviates splenomegaly or bladder enlargement.

16. The pharmaceutical composition according to claim 1, which increases the expression of ZO-1 and occludin in urothelium.

17. The pharmaceutical composition according to claim 1, which protects urothelial cells.

18. The pharmaceutical composition according to claim 1, wherein the inflammatory skin disease is an autoimmune skin disease, a proliferative skin disease, a fibrous skin disease, contact dermatitis, seborrheic dermatitis, sclerosis cutaneously, hypersensitivity to skin, atopic dermatitis, urticaria, pruritus, psoriasis, or eczema.

19. The pharmaceutical composition according to claim 18, wherein the psoriasis is guttate psoriasis, plaque psoriasis, erythrodermic psoriasis, pustular psoriasis, and exfoliative psoriasis.

20. The pharmaceutical composition according to claim 1, which inhibits or alleviates erythema of the skin, scaling of the skin, or thickening of the epidermal tissue.

21. A composition for improving bladder dysfunction, comprising a compound of chemical formula 1, its isomer, its solvate, its hydrate, or its salt as an active ingredient. 【Chemistry 1】

22. The composition for improving bladder dysfunction according to claim 21, wherein the bladder dysfunction is bladder dysfunction caused by spinal cord injury.

23. The composition for improving bladder dysfunction according to claim 22, wherein the spinal cord injury is traumatic spinal cord injury, degenerative spinal disease, inflammatory spinal disease, spinal cord tumor, spinal cord malformation, spinal tumor, spinal cord hemorrhage, stroke, spinal cord paralysis due to extramedullary vascular disease, myelitis, multiple sclerosis, amyotrophic lateral sclerosis, or spinal stenosis.

24. A composition for improving bladder dysfunction according to claim 21, which improves or restores the urinary function of the bladder.

25. A composition for inhibiting bladder enlargement, comprising a compound of chemical formula 1, its isomer, its solvate, its hydrate, or a salt thereof as an active ingredient. 【Chemistry 1】

26. The bladder enlargement inhibitory composition according to claim 25, which suppresses the thickening of the urothelial layer, lamina propria, or muscular layer of the bladder.

27. A cosmetic composition for the prevention or improvement of inflammatory skin diseases, comprising a compound of chemical formula 1, its isomer, its solvate, its hydrate, or its salt as an active ingredient. 【Chemistry 1】

28. The cosmetic composition for preventing or improving inflammatory skin diseases according to claim 27, manufactured in one or more dosage forms selected from the group consisting of lotion, cream, serum, emulsion, gel, hand cream, lipstick, cleansing foam, cleansing cream, cleansing water, spray, shampoo, conditioner, treatment, body cleanser, soap, pack, massage agent, face powder, compact, foundation, two-way cake, and makeup base.

29. Uses of the compound of chemical formula 1, its isomers, its solvates, its hydrates, or its salts for use in the manufacture of pharmaceutical compositions for the prevention or treatment of neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin diseases. 【Chemistry 1】

30. Uses of the compound of chemical formula 1, its isomers, its solvates, its hydrates, or its salts for use in the manufacture of compositions for improving bladder dysfunction or suppressing bladder enlargement. 【Chemistry 1】

31. A method for treating neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin disease, comprising the step of administering a compound of chemical formula 1, its isomer, its solvate, its hydrate, or a salt thereof to an individual suffering from neuropathic pain, neuroinflammation, bladder disease, neurogenic bladder, or inflammatory skin disease. 【Chemistry 1】

32. A method for treating or improving bladder dysfunction or bladder enlargement, comprising the step of administering a compound of chemical formula 1, its isomer, its solvate, its hydrate, or a salt thereof to an individual having symptoms of bladder dysfunction or bladder enlargement. 【Chemistry 1】

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