JP6426746B2 - 4-Methylsulfonyl-2-butenenitrile and its pharmaceutical use - Google Patents

Description

FIELD OF THE INVENTION The present invention relates to 4-methylsulfonyl-2-butenenitrile and pharmaceutical compositions comprising the same. The invention also relates to methods of using the compounds to treat inflammation or inflammation related disorders and pain.

BACKGROUND OF THE INVENTION Inflammation is the process by which microbe or tissue damage induces the release of cytokines and chemokines from various cell types, increased vascular permeability, upregulation of endothelial receptors and thus the birth of various cells. It results in an increase in the exit of the sexual and adaptive immune system, enters the surrounding tissue and largely produces the traditional state of inflammation, ie, redness, swelling, fever and pain.

  Inflammation is the local response of living tissue to damage that may be caused by various endogenous and exogenous factors. Exogenous factors include physical, chemical and biological factors. Endogenous agents include inflammatory mediators, antigens and antibodies. Intrinsic factors often appear under the influence of exogenous damage. The inflammatory response often then results in altered structure and permeability of the cell membrane. Endogenous factors such as mediators and antigens determine the type or type of inflammatory response, in particular the course of the inflammatory response in the area of injury. An acute form of inflammation occurs when tissue damage is limited to the production of mediators. If the immune response is involved in the process, a long-term inflammatory process emerges through the interaction of antigens, antibodies and autoantigens. For example, various extrinsic natural forces such as infections, trauma, radiation also provide an inflammatory process at the molecular level by damaging the cell membrane which initiates the biochemical reaction.

  Based on physical factors, pain can be divided into three types: nociceptive, neuropathic and mixed.

  Nociceptive pain is a term relating to pain detected by nociceptors. Nociceptors are free nerve endings that terminate directly under the skin, in the tendon, in the joint and in the viscera. Nociceptive pain typically responds well to treatment with opioids and NSAIDs. There are several types of nociceptive pain: somatic pain, visceral pain and skin pain. Visceral pain comes from the viscera. Deep somatic pain is a dull, itchy, poorly localized pain that is initiated by stimulation of nociceptors in the ligaments, tendons, bones, blood vessels, fascia and muscles. Examples include sprains and fractures. Superficial pain is initiated by the activation of nociceptors in the skin or other surface tissue and is in a sharp, clear, well-defined position. Examples of injuries causing superficial somatic pain include minor and minor burns (first degree). Nociceptive pain is usually short in duration and ends when the injury recovers. Examples of nociceptive pain include post-operative pain, sprains, bone fractures, burns, bumps, bruises, and inflammatory nociceptive pain. Inflammatory nociceptive pain is associated with tissue damage and the resulting inflammatory process.

  Neuropathic pain is caused by damage to neurons in the peripheral and central nervous systems, accompanied by sensitization of these systems. Because the underlying etiology is usually irreversible, most neuropathic pain is chronic pain. Most people express neuropathic pain as shooting, burning, tingling, blunting, electric shock, numbness and persistent allodynia. The nomenclature of neuropathic pain is based on the etiology and sites that initiate the nervous system, for example, post-stroke pain, diabetic peripheral neuropathy, post-herpetic (or post-zoster) neuralgia, terminal cancer pain, phantom pain It is limb pain.

  Mixed pain is characterized by the coexistence of both nociceptive and neuropathic pain. For example, myalgia induces central or peripheral nerve sensitization leading to chronic low back pain, migraine headache and myofascial pain.

  Connective tissue is exposed to constant stress and injury. Acute or chronic impact and the natural progression of various degenerative diseases all cause painful inflammation in joint areas such as the neck, back, arms, hips, ankles and feet. These pains are common and often debilitating.

  There is a need for compositions and methods for treating inflammation, inflammation related disorders and pain. The composition should be economical and easy to manufacture, and the method should be effective and have no significant side effects.

SUMMARY OF THE INVENTION The present invention relates to 4-methylsulfonyl-2-butenenitrile or a pharmaceutically acceptable salt thereof, one isomer or multiple isomers. The present invention also relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and 4-methylsulfonyl-2-butenenitrile or a pharmaceutically acceptable salt thereof, or a solvate thereof. The compound is preferably at least 90% pure (w / w).

  The invention also relates to methods of treating inflammation and pain. The method comprises the step of administering 4-methylsulfonyl-2-butenenitrile, or a pharmaceutically acceptable salt thereof, to the subject in need. Pharmaceutical compositions containing the active compounds can be applied by any of the accepted modes of administration, including topically, orally and parenterally (e.g. intravenously, intramuscularly, subcutaneously or rectally and the like). Topical administration and oral administration are preferred.

Detailed Description of the Invention As used herein, "about" refers to ± 10% of the stated value.

"Pharmaceutically acceptable salt" as used herein is a salt that retains the desired biological activity of the present compounds and does not impart undesired toxic effects. Pharmaceutically acceptable salt forms include various crystalline polymorphs as well as amorphous forms of different salts. Pharmaceutically acceptable salts may be formed with metal or organic counterions and include, but are not limited to, alkali metal salts such as sodium or potassium, alkaline earth metal salts such as magnesium or calcium, and ammonium or tetraalkyl Ammonium salts may be mentioned, i.e. NX4 ⁺ (X is _C1-4 ).

  "Solvate" as used herein is an addition complex in which a compound is combined with an acceptable cosolvent at a particular fixed rate.

4-Methylsulfonyl-2-butenenitrile We have isolated and identified 4-methylsulfonyl-2-butenenitrile. The compound or a pharmaceutically acceptable salt or solvate thereof is effective for treating inflammation, inflammation related disorders and pain.

  4-Methylsulfonyl-2-butenenitrile has a molecular weight of 145.18, and its trans structure is shown below.

4-Methylsulfonyl-2-butene nitrile

  4-Methylsulfonyl-2-butenenitrile can be synthesized by the addition of sodium methanesulfinate to 4-bromo-2-butenenitrile in an organic solvent containing water, such as aqueous ethanol. The required 4-bromo-2-butene nitrile brominates allyl cyanide to provide 3,4-dibromobutane nitrile, then base catalyzed removal of HBr to form a double bond between C2 and C3 It can be prepared by providing and is expected to be an approximately 1: 1 mixture of E- and Z-isomers.

  The present invention relates to a process for the preparation of 4-methylsulfonyl-2-butenenitrile. The process comprises the step: (a) reacting -bromo-2-butenenitrile with sodium methanesulfinate. Any solvent or solvent mixture that is suitable to provide a homogeneous solution of reactants is suitable. For example, water miscible organic solvents such as ethanol, methanol, isopropanol, acetone, tetrahydrofuran, or dioxane, containing a small amount of water (1 to 10%, or 1 to 5%) are suitable solvents for the reaction. In one embodiment, ethanol: water in a ratio of 30 to 40: 1 is used as a solvent for the reaction.

The process comprises firstly reacting bromine (Br ₂ ) with allyl cyanide prior to step (a) above and then adding the basic solution to prepare the required 4-bromo-2-butene nitrile ( i) may further be included. In this process, allyl cyanide is first brominated in an organic solvent such as petroleum ether and alcohol and then by introducing at least one equivalent, preferably about one equivalent of a suitable base The HBr is removed from the molecule to form a double bond. In this step, a suitable base that is compatible with the solvent in the reaction mixture is used. The preferred base is an ethanolic solution of sodium ethoxide, which can be prepared by dissolving sodium metal in ethanol. After isolation and purification of the product 4-bromo-2-butenenitrile, it may be used in step (a).

Pharmaceutical Composition The present invention provides a pharmaceutical composition comprising one pharmaceutically acceptable carrier and an active compound of 4-methylsulfonyl-2-butenenitrile or a pharmaceutically acceptable salt thereof or a solvate thereof. . The pharmaceutical composition can comprise both isomers in one or equimolar amounts of cis or trans, or in different amounts. The "active compound" or a pharmaceutically acceptable salt or solvate thereof in a pharmaceutical composition is generally about 0.01-20% or 0.05-20% or 0.1-0.1% in a topical preparation. 20% or 0.2 to 15% or 0.5 to 10% or 1 to 5% (w / w) amount, about 0.1 to 5% for injectable formulation, 0. 0 for patch formulation. 1 to 5%, about 1 to 90% for tablet formulations and 1 to 100% for capsule formulations.

  In one embodiment, the active compound is incorporated in any acceptable carrier, such as a cream, gel, lotion or other that can stabilize the active compound and deliver it to the affected area by topical application. Types of suspensions include: In another embodiment, the pharmaceutical composition can be in the form of tablets, capsules, granules, fine granules, powders, syrups, suppositories, injectable solutions, patches and the like. The above pharmaceutical compositions may be prepared by conventional methods.

  Pharmaceutically acceptable carriers that are inactive ingredients may be selected by one skilled in the art using conventional criteria. Pharmaceutically acceptable carriers include, but are not limited to, non-aqueous solutions, suspensions, emulsions, microemulsions, micellar solutions, gels and ointments. Pharmaceutically acceptable carriers may also include the following components, including, but not limited to, saline and aqueous electrolyte solutions; ionic such as sodium chloride, potassium chloride, glycerol and dextrose and Nonionic penetrants; pH adjusters and buffers such as hydroxide salts, phosphates, citrates, acetates, borates and trolamines; bisulfites, sulfites, metabisulfites, thios Sulfites, ascorbic acid, acetylcysteine, cysteine, glutathione, butylhydroxyanisole, butylated hydroxytoluene, tocopherol and salts of ascorbyl palmitate, acids and / or bases, and other antioxidants; surfactants such as lecithin and phospholipids (But not limited to, phosphatidyl choline, phosphatidyl ether, Poloxamer and poloxamine, polysorbate 80, polysorbate 60 and polysorbate such as polysorbate 20, polyethers such as polyethylene glycol and polypropylene glycol; polyvinyl compounds such as polyvinyl alcohol and povidone; methyl cellulose, hydroxypropyl cellulose Cellulose derivatives such as hydroxyethyl cellulose, carboxymethyl cellulose and hydroxypropyl methyl cellulose and salts thereof; petroleum derivatives such as mineral oil and white petrolatum; fats such as lanolin, peanut oil, palm oil, soybean oil; mono-, di- and triglycerides; Acrylic acid polymers such as polymethylene gel and hydrophobically modified crosslinked acrylate copolymers Mar; include polysaccharides such as glycosaminoglycans, such as dextran and sodium hyaluronate. Such pharmaceutically acceptable carriers can be preserved against bacterial contamination using well known preservatives, where such preservatives include, but are not limited to, benzalko chloride And ethylenediaminetetraacetic acid and salts thereof, benzethonium chloride, chlorhexidine, chlorobutanol, methyl paraben, thimerosal and phenylethyl alcohol, or formulated as a non-preserved preparation either single or multiple use it can.

  For example, tablet or capsule formulations of the active compound have no bioactivity and can include other excipients that do not have the reactivity of the active compound. The excipients for tablets or capsules can include fillers, binders, lubricants and glidants, disintegrants, wetting agents and release rate modifiers. Binders promote the adhesion of the particles of the formulation and are important for tablet formulations. Examples of excipients for tablets or capsules include, but are not limited to, carboxymethylcellulose, cellulose, ethylcellulose, hydroxypropyl methylcellulose, methylcellulose, karaya gum, starch, gum tragacanth, gelatin, magnesium stearate, titanium dioxide, poly ( Acrylic acid) and polyvinyl pyrrolidone. For example, the tablet formulation can include inactive ingredients such as colloidal silicon dioxide, crospovidone, hypromellose, magnesium stearate, microcrystalline cellulose, polyethylene glycol, sodium starch glycolate and / or titanium dioxide. Capsule formulations may include inactive ingredients such as gelatin, magnesium stearate and / or titanium dioxide.

  For example, patch formulations of the active compound may be 1,3-butylene glycol, dihydroxyl aluminum aminoacetate, disodium edetate, D-sorbitol, gelatin, kaolin, methyl paraben, polysorbate 80, povidone, propylene glycol, propyl paraben, carboxymethylcellulose sodium It can contain several inactive ingredients such as sodium polyacrylate, tartaric acid, titanium dioxide and purified water. Patch formulations may also include skin permeation enhancers such as lactate esters (eg, lauryl lactate) or diethylene glycol monoethyl ether.

  Topical formulations containing the active compound can be in the form of gels, creams, lotions, liquids, emulsions, ointments, sprays, solutions and suspensions. Inactive ingredients in topical preparations include, but are not limited to, for example, lauryl lactate (emollient / permeability enhancer), diethylene glycol monoethyl ether (emollient / permeability enhancer), DMSO (dissolution) Accelerators), silicone elastomers (rheology / texture modifiers), caprylic acid / capric acid triglycerides, (emollients), octyl salicylate (octisalate) (emollients / UV filters), silicone fluids (emollients / diluents) And squalene (emollients), sunflower oil (emollients) and silicon dioxide (thickeners).

  In one embodiment, lauryl lactate (eg, about 0.1 to 10% or about 0.2 to 5% or 0.5 to 5%) is included in the topical gel formulation. Lauryl lactate is considered safe for topical administration. Lauryl lactic acid is suitable for human use in medicine and cosmetics. Lauryl lactate enhances the permeability of compounds when used in topical formulations. Preferably, lauryl lactate is purified to achieve a purity of 9090%, preferably ≧ 95%, the high purity reducing the presence of hydrolysing agents and oxidizing agents. Also, 0.1-20% or 0.5% -10% (w / w) DMSO in the formulation provides adequate solubility of the active compound.

  In another embodiment, diethylene glycol monoethyl ether is included in the topical gel formulation.

Methods of Use Inflammation is a process and histopathologic condition resulting from the continuation of the activation and activation of the innate and acquired components of the immune system. The arachidonic acid cascade and cytokine production and action in cell-cell interactions are important elements of immune activation and response that cause inflammation. Arachidonic acid is a component of membrane phospholipids. After arachidonic acid is released from phospholipids, arachidonic acid acts as a precursor of many known eicosanoids, including prostaglandins and leukotrienes, which are known proinflammatory entities.

  The active compound is anti-inflammatory when applied topically in a mouse ear swelling model, where inflammation is induced by arachidonic acid. The active compounds are effective to inhibit proinflammatory mediators.

  The present invention relates to a method of treating inflammation and / or pain. 4-Methylsulfonyl-2-butenenitrile may be used as it is or may be administered in the form of a pharmaceutical composition additionally comprising a pharmaceutically acceptable carrier. The method comprises the steps of first identifying a subject afflicted with inflammation and / or pain, and administering to the subject an active compound in an amount effective to treat inflammation and / or pain. An "effective amount," as used herein, is an amount effective to treat a disease by ameliorating the condition or reducing the symptoms of the disease.

  In one embodiment, the method reduces or ameliorates a symptom associated with inflammation. The present invention provides a method of treating local symptoms of inflammation characterized by acute or chronic swelling, pain, redness, elevated temperature or loss of function in some cases.

  In another embodiment, the present invention provides a method of ameliorating the symptoms of pain regardless of the cause of the pain. The general term "pain" treatable by this method includes nociceptive, neuropathic and mixed. The present invention reduces pain of varying severity, ie, mild, moderate and severe pain, acute and chronic pain. The present invention is effective in treating arthralgia, myalgia, pain in tendons, pain caused by burns, and pain caused by inflammation such as rheumatoid arthritis.

  In one embodiment, the present invention is useful for treating inflammation and / or pain associated with the musculoskeletal system or skin. The highly innervated musculoskeletal and skin systems have a high ability to display pain. Also, the musculoskeletal system has a high capacity for tissue enlargement, and the skin has a high capacity for redness, swelling and heat. In the musculoskeletal and skin systems, the degree of tissue damage often increases in proportion to the inflammatory response that occurs. For example, in skin, mere firm rubbing will cause the release of cytokines, IL-1 and TNF.

  The present invention provides methods of treating pain associated with inflammation and / or inflammatory skeletal or muscle diseases or conditions. The method comprises the steps of identifying a subject in need thereof and administering the active compound to the subject in an amount effective to treat inflammation and / or pain. "Active compound", as used herein, is intended to include 4-methylsulfonyl-2-butenenitrile and its pharmaceutically acceptable salts, one isomer or multiple isomers thereof . Skeletal or muscle disease or condition is musculoskeletal sprain, musculoskeletal contusion, tendon disorder, peripheral nerve root disease, osteoarthritis, joint degenerative disease, polymyalgia rheumatica, juvenile arthritis, gout, ankylosing spondylitis, psoriatic Arthritis, systemic lupus erythematosus, costochondritis, tendonitis, bursitis, eg, normally lateral epicondylitis (tennis elbow), medial epicondylitis (pitcher elbow) and trochanteric bursitis, temporomandibular joint syndrome and Fibromyalgia may be mentioned.

  The present invention provides methods of treating inflammation and / or inflammatory skin diseases such as dermatitis, psoriasis and pain associated with acne. The method comprises the steps of identifying a subject in need thereof and administering to the subject an effective amount of the active compound to treat inflammation and / or pain.

  The skin is highly responsive to environmental stimuli, and the epidermal component of keratinocytes is a very rich source of both arachidonic acid and the proinflammatory cytokines IL-1 and TNF. Dermal dendritic cells, Langerhans cells, recognize and process antigens for additional immune responses of various lymphocytes, and all of these cells are cytokines via their specific cell surface receptors Mainly adjusted by

  Dermatitis (also called eczema) is a common inflammation of the skin. Specific types of dermatitis include atopic, tactile, money-like and light-induced.

  4-Methylsulfonyl-2-butenenitrile, which is effective in inhibiting arachidonic acid-induced inflammation and effective in inhibiting proinflammatory mediators, can be used to treat inflammation and / or psoriasis, acne, rosacea, It is also effective for treating the pain associated with dermatitis, especially contact dermatitis and atopic dermatitis.

  The pharmaceutical composition of the present invention can be applied by local administration and systemic administration. Local administration includes topical administration. Systemic administration includes oral, parenteral (including intravenous, intramuscular, subcutaneous or rectal) and other systemic routes of administration. On systemic administration, the active compound first reaches the plasma and is then distributed in the target tissue. Topical and oral administration are preferred routes of administration for the present invention.

The dosage of the composition can vary based on the degree of injury and the response of each individual patient. For systemic administration, the concentration in plasma of the active compound to be delivered may vary, but in general 1 × 10 ^-10 to 1 × 10 ^-4 moles / liter, preferably 1 × 10 ^-8 to 1 × 10 ^-5 It is mol / liter.

  In one embodiment, the composition is topically applied to the affected area and rubbed in. The composition is applied topically at least once or twice a day, or 3 to 4 times a day, depending on the medical problem and the disease condition being chronic or acute. Generally, the topical composition is about 0.01 to 20% or 0.05 to 20% or 0.1 to 20% or 0.2 to 15%, 0.5 to 10 or 1 to 5% (w / w) The active compound of For example, the topical composition comprises about 1 or 5% (w / w) of active compound. Depending on the size of the affected area, 0.2 to 85 mL, typically 0.2 to 10 mL of the topical composition is applied to the individual per dose. The active compound passes through the skin and is delivered to the site of discomfort.

  In one embodiment, the pharmaceutical composition is orally administered to the subject. The dosage for oral administration is generally 1 to 50, preferably 1 to 10 or 1 to 5 mg / kg / day. For example, the active compound may be applied orally to adults at 100 to 800 mg / dose or 200 to 600 mg / dose 1-4 times / day, depending on the patient's condition.

  In one embodiment, the pharmaceutical composition is administered subcutaneously to the subject. The dosage for subcutaneous administration is generally 0.3 to 20, preferably 0.3 to 3 mg / kg / day.

  One skilled in the art will recognize that various types of delivery mechanisms are also suitable for the present invention.

  The invention is useful for treating mammalian subjects such as humans, horses and dogs. The invention is particularly useful for treating humans.

  The following examples further illustrate the invention. These examples are intended merely to illustrate the present invention and should not be construed as limiting.

Example 1 Preparation of 4-Methylsulfonyl-2-Butene Nitrile (A) A solution of allyl cyanide (0.59 mol) in t-amyl alcohol (120 mL) and petroleum ether (370 mL), sequentially with t-amyl alcohol (60 mL) The solution was treated with a solution of bromine (0.59 mol) in) and then with an ethanolic solution of sodium ethoxide (345 mL, 0.60 mol). When the reaction mixture was cooled to room temperature, the solids were removed by vacuum filtration and the filtrate was concentrated under reduced pressure. The residual liquid was loaded onto a silica gel column (307.10 g) and eluted with hexane-ethyl acetate (19: 1; 9: 1). The appropriate fractions were combined and then concentrated under reduced pressure to provide 4-bromo-2-butenenitrile as a pale yellow liquid (47.14 g).

(B) The product from part (A) (150 mmol) is added dropwise over about 14 to an equimolar solution of sodium methanesulfinate in ethanol (120 mL) containing water (4 mL), about 40 ° C. Stir. After 6 hours of continuous heating and stirring, the reaction mixture was concentrated under reduced pressure. The dark residual syrup was diluted with acetone and then vacuum filtered to remove sodium bromide. The filtrate was concentrated to give a dark brown syrup which was loaded onto a silica gel column (about 255 g) and eluted with chloroform-ethanol (19: 1). Combination of the appropriate fractions followed by concentration under reduced pressure gave a pale amber liquid (16.08 g). The liquid (10.48 g) was loaded onto a silica gel column (about 310 g) and eluted with chloroform-ethanol-hexane (66: 4: 30). Combination of the appropriate fractions followed by concentration under reduced pressure gave a pale amber liquid which was taken in acetone and degranulated with a 0.45 μm syringe filter. Concentration of the clear filtrate gave the expected product (9.71 g). The ¹ H NMR spectrum of 4-methylsulfonyl-2-butenenitrile was acquired by Spectral Data Services, Inc. at 400 MHz in DMSO-d ₆ solution. Table 1 shows chemical shift data (ppm) of NMR results.

The structure of 4-methylsulfonyl-2-butenenitrile contains seven protons. All protons cause rational chemical shifts and occur there. The assignment of chemical shifts for methyl and methylene protons is clear. Two groups of multiplets at 6 ppm and 6.2 ppm can be identified by the coupling constant (J). J-16 Hz is consistent with the expected coupling constant for alkene protons in the E structure, and J-11 Hz is consistent with the expected coupling constant for alkene protons in the Z structure. Based on a comparison of the RIV values of these signals, the E / Z ratio is about 58/42. The coupling constants of the remaining alkene protons can not be accurately measured due to the complexity of the signal between 6.6 and 7 ppm. The spectrum contains the impurity / solvent peak (RlV _Total 3.75).

Example 2 Gel formulation 1
Table 2 illustrates one gel formulation comprising 4-methylsulfonyl-2-butenenitrile.

Example 3. Anti-inflammatory activity of active compound by topical application in mice 4-Methylsulfonyl-2-butenenitrile prepared from Example 1 was used in this experiment.
The anti-inflammatory activity was evaluated in a local arachidonic acid-induced ear swelling model in mice for test compounds, indomethacin (positive control) and vehicle.
Male ICR mice weighing 22 ± 2 g were randomly divided; test compounds and vehicle controls had 10 mice and indomethacin had 5 mice. Arachidonic acid (0.5 mg in 20 μl acetone: ethanol 1: 1) was topically applied to the anterior and posterior surfaces of the right ear of each mouse. The test substance and vehicle were applied similarly 30 minutes before and 15 minutes after arachidonic acid application as shown in Table 2. The thickness of the right and left ears was measured and the difference was calculated as an indicator of right ear inflammation. Ear swelling was measured with a Dyer model micrometer gauge 60 minutes and 90 minutes after application of arachidonic acid as an indicator of inflammation. Percent inhibition was calculated by the formula: Ic-It / Icx 100, where Ic and It refer to the increase in ear thickness (mm) in control and treated mice, respectively. ANOVA and Durnnett tests were used to confirm significant differences between vehicle control and treated groups. Significance is set at p <0.05 level. The results after 90 minutes of arachidonic acid application are shown in Table 2.

  Test compounds resulted in 22% inhibition of arachidonic acid-induced ear swelling compared to the vehicle treated group. The difference between treated and vehicle treated mice was determined to be statistically significant (p-value by t-test was 0.002).

Example 4 Anti-inflammatory activity of active compounds in mice by oral application (predictive example)
The active compound 4-methylsulfonyl-2-butenenitrile is suspended at 5 to 15 mg / mL in vehicle (DMSO). Test compounds, dexamethasone (positive control) and vehicle are orally administered to mice and anti-inflammatory activity is assessed in a local arachidonic acid-induced ear swelling model in mice.

  Male ICR derived mice are used in this experiment. Ten mice are used for each group (active compound, positive control and vehicle). All animals are maintained in a controlled temperature (22-24 ° C.) and humidity (60% -70%) environment with a 12 hour light / dark cycle for at least 1 week prior to use.

  Arachidonic acid (0.5 mg in 20 μl of acetone) is topically applied to the anterior and posterior surfaces of the right ear of the test animal. The test substance (10 mL / kg) and vehicle (10 mL / kg, 50 to 150 mg / kg) in vehicle are orally administered by gavage one hour before arachidonic acid application, while dexamethasone is loaded with arachidonic acid Three hours prior to administration orally by gavage. The thickness of the right and left ears is measured 60 and 90 minutes after arachidonic acid induction of ear edema, and the difference is calculated as an indicator of right ear inflammation. Significant activity is defined as statistically significant inhibition (as determined by t-test, p-value <0.05) in arachidonic acid-induced ear swelling compared to the vehicle treated group.

Example 5 Analgesic activity of the active compound by oral administration in mice (formalin model, predictive example)
The formalin test is a model of continuous pain resulting from formalin induced tissue damage. Nociceptive pain and inflammatory pain are induced by injection into the foot of dilute formalin solution, resulting in nociceptive behavior including foot contraction. The formalin model involves inflammation of pain, neurogenesis and central mechanisms. The early stages of pain (0 to about 10 minutes) are due to nociceptive mechanisms, and the late stages of pain (10 to 40 minutes) are due to a combination of inflammatory pain and nociceptive mechanisms. Pain behavior is assessed using forefoot licking measurements. The final point of the study is the number of foot licking events (Hunskaar et al., Pain, 30: 103-114, 1987; Li et al., Molecular Pain, 6: 11, 2010).
Ten mice per group are used in this study. Just prior to testing (time 0), mice are restrained in cloth and 20 μL of 5% formalin solution is injected subcutaneously into the back surface of the left hind paw. Vehicle control (DMSO) and test compound 4-methylsulfonyl-2-butenenitrile (in DMSO) are orally administered to mice in a volume of 5 mL / kg by gavage. The amount of test compound is 100 or 500 mg / kg / dose.

  The positive control morphine in saline is administered by subcutaneous injection into mice at 8 mg / kg immediately prior to formalin injection and tested at time zero.

  Following formalin injection, animals are placed in individual cages and manually observed for 60 minutes. The licking events are recorded at 5 minute intervals and lasts for a total of 60 minutes.

  The number of licking events at different time points after formalin injection of vehicle control, morphine-treated and test compound-treated mice are plotted at 5-minute intervals. The number of licking events per minute is calculated between 0-10 minutes and 10-40 minutes for vehicle, positive control and test compound. A statistically significant decrease in licking events per minute is an indicator that the test compound is effective in treating acute nociceptive pain (early phase) or inflammatory nociceptive pain (late phase) .

Example 6 Analgesic activity of active compounds by topical administration in mice (formalin model, predictive example)
The animals and treatment procedures are similar to those described in Example 5 with the following exceptions.
The test compound 4-methylsulfonyl-2-butenenitrile (375 mM in vehicle, n = 10) and vehicle control (acetone: ethanol 1: 1, n = 10) sink the mouse left hind leg in each solution for about 30 seconds Administration locally. Then remove the foot and wipe with tissue to avoid excessive skin dryness.

  The positive control morphine is administered by subcutaneous injection at 8 mg / kg in saline (n = 10).

  Morphine is given subcutaneously once 15 minutes before formalin injection. Test compounds and vehicle controls are topically administered twice 60 minutes prior to formalin injection (BID).

  Following formalin injection, animals are placed in individual cages and manually observed for 60 minutes. The licking events are recorded at 5 minute intervals and lasts for a total of 40 minutes.

The number of licking events at different time points after formalin injection of vehicle control, morphine treated and test compound treated mice is determined.
The number of licking events per minute is calculated between 0-10 minutes and 10-40 minutes for vehicle, positive control and test compound. Two sample t-tests are performed to compare the vehicle group to the test compound group. Significance is set at p <0.05 level.

Example 7 Analgesic activity of active compounds in chronic contraction injury model (predictive example)
Peripheral nerve lesions can cause pathological phenomena including spontaneous pain as well as exaggerated responses to light touch (touch allodynia). The chronic stenosis injury model is a neuropathic pain model.
Use male Sprague Dawley rats. Under anesthesia with pentobarbital (50 mg / kg, 5 ml / kg, ip) the sciatic nerve is exposed at the mid-thigh level. Loosely connect four ligatures (4-0 chrom gut) about 1 mm apart around the nerve. Animals are then individually placed in cages with soft bedding for 7 days prior to testing. Narrowing of the sciatic nerve causes nerve damage and unilateral neuropathic pain.

  On the day of the experiment, the animals do not have access to food overnight prior to testing. The rat is placed on a wire mesh rack under an inverted plexiglass cage and allowed to acclimate for 20-30 minutes. Mechanics Allodynia is assessed by the Chapplan up / down method using von Frey filaments on the plantar surface of the left hind paw. See Chaplan, et al. J. Neuroscience Methods, 53: 55-63, 1994.

  In rats, the pain threshold 7-14 days after nerve ligation (pretreatment) is reduced by 10 g of force before the nerve ligation (before ligation), that is, there is a clear allodynia Only if preselected for the experiment.

  The active compound 4-methylsulfonyl-2-butenenitrile is prepared in a gel formulation according to example 2.

  Active compound in gel formulation (1 or 5% topical administration), active compound in DMSO (500 mg / kg, oral administration), morphine (positive control, subcutaneously, 8 mg / kg in physiological saline), topical vehicle (active compound) And gel vehicle (DMSO) without

  The test substance or vehicle is administered orally or topically on the plantar surface of the left hind paw. The mechanical allodynia test is performed 30 minutes before (pretreatment) and 1 and 3 hours after a single dose of test substance or vehicle (post treatment). The paw withdrawal thresholds of control and test compounds are measured.

Example 8 Treatment of knee pain by local administration (predicted example)
Aim: To investigate the efficacy of active compounds in topical gel formulations in human patients with moderate to severe knee pain associated with osteoarthritis after temporary discontinuation of standard NSAID therapy. This study focuses on the pathological phenomena caused by painful arthritis. Clinical trials use knee osteoarthritis as a well established paradigm for other musculoskeletal injuries.

  Topical Formulation: A gel formulation containing 1% and 5% 4-methylsulfonyl-2-butenenitrile (Example 2) is used in this example. The placebo comprises the same gel without the active compound.

  Methods: Randomized double-blind placebo-controlled parallel-centered multicenter clinical activity study (A randomized, double-blind, placebo controlled, parallel treatment multicenter clinical activity study)

  Patients with knee pain osteoarthritis controlled by a stable dose of standard NSAID therapy for at least 2 months stop using the NSAID for a 7 day washout period. Patients are then randomized at a ratio of 1: 1: 1 (1% active gel, 5% active gel, placebo). A total of 150 patients will be enrolled.

  The active gel or placebo is applied to the affected knee three times a day for 12 weeks, for a total of 252 treatments taking place every 4 to 6 hours.

  Patients are treated for 12 weeks and followed up for 4 more weeks. The NSAID can be restarted for evaluation after the 12th week visit.

Safety criteria for evaluation · Adverse events (AE) throughout the study
Physical examination at entry (day -7, start of NSAID without withdrawal), Baseline (day 1, start of treatment), Week 12 and Week 16-Vital sign at entry (day-7, Start of NSAID without withdrawal), baseline (day 1, start of treatment) and weeks 2, 4, 6, 12 weeks and week 16 baseline (day 1), 4, 8, 12 and Clinical laboratory measurements at 16.

Clinical activity:
The primary clinical activity parameter is a measure of pain in the target joint as quantified by Visual Analogue Scale (VAS) and Western Ontario and McMaster University (WOMAC) Index Pain Subscale. The effects on knee swelling, tenderness and inflammation are recorded, and the time to reduction or eradication of pain after treatment is recorded.

Study end point The primary clinical activity end point is
WOMAC Dysfunction Index The change from baseline (day 1) to week 12 (scale 0-20) in the pain subscale.
Secondary clinical activity end point is
Change from baseline (day 1) to week 12 in WOMAC dysfunction index pain subscale stiffness (scale 0-8)
・ Physical function (scale 0 to 68)
Change from baseline (day 1) to week 12 in the VAS pain score (scale 0-100)
Change from baseline (day 1) to week 2 in the VAS pain score (scale 0-100)
Change in investigators' assessment of swelling, tenderness and inflammation at baseline (Day 1) and Weeks 4 and 12 after the first application on Day 1 Active gel or placebo gel Pain reduction or time to eradication after each topical application of each • Emergency drug (APAP) use.

Example 9 Treatment of knee pain by oral administration (predicted example)
The design and protocol of this experiment are similar to those described in Example 9, but the active compound and placebo are applied by oral route.
Oral formulations: Tablet formulations containing 10, 100 or 1000 mg of active compound 4-methylsulfonyl-2-butenenitrile are used in this example. The placebo is the same tablet formulation without the active compound.

Methods Patients are randomized at 1: 1: 1: 1 (10 mg: 100 mg: 1000 mg: placebo). Include up to 200 patients in total.
A total of 168 treatments are given orally to each patient, giving active tablets or placebo twice a day for 12 weeks, awake every 12 hours. Patients are treated for 12 weeks and followed up for another 4 weeks.
The criteria for the evaluation are identical to those described in Example 9.

  The foregoing describes the preferred embodiments of the present invention, and it should be understood that modifications can be made within the scope of the present invention as set forth in the claims.

Claims (14)

  A compound of 4-methylsulfonyl-2-butenenitrile or a pharmaceutically acceptable salt thereof.   A pharmaceutical composition comprising a pharmaceutically acceptable carrier, and the compound according to claim 1 or a pharmaceutically acceptable salt thereof. Has a purity of the compound is at least 90% (w / w), and wherein the composition is a topical form of gels, creams, lotions, ointments, solutions, suspensions, spray or patch, according to claim 2 Pharmaceutical composition. The pharmaceutically acceptable carrier is an emollient selected from the group consisting of lauryl lactate, diethylene glycol monoethyl ether, caprylic / capric acid triglyceride, octyl salicylate (octisalate), silicone fluid, squalene and sunflower oil. The pharmaceutical composition as described in 2 .   The pharmaceutical composition according to claim 2, wherein the pharmaceutically acceptable carrier is diethylene glycol monoethyl ether.   The pharmaceutical composition according to claim 5, further comprising acrylate / C10-30 alkyl and tris (2-hydroxyethyl) amine.   The pharmaceutical composition according to claim 2, wherein the compound has a purity of at least 90% (w / w) and the composition is in oral form in tablets, capsules, granules, powders or syrups.   A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof in an amount effective to treat inflammation or pain, for treating inflammation or pain.   9. The pharmaceutical composition according to claim 8, for reducing or alleviating signs of local symptoms of inflammation characterized by acute or chronic swelling, pain or redness.   The inflammation and / or pain may be musculoskeletal sprain, musculoskeletal contusion, tendon disorder, peripheral radiculopathy, osteoarthritis, joint degenerative diseases, juvenile arthritis, gout, ankylosing spondylitis, psoriatic arthritis, systemic lupus erythematosus, The pharmaceutical composition according to claim 8, which is associated with a skeletal or muscle disease or condition selected from the group consisting of costochondritis, tendinitis, bursitis, temporomandibular joint syndrome and fibromyalgia.   9. The pharmaceutical composition according to claim 8, wherein the inflammation and / or pain is associated with a joint, a ligament, a tendon, a bone, a muscle or a fascia.   9. The pharmaceutical composition according to claim 8, wherein the inflammation and / or pain is associated with an inflammatory skin disease or disorder of dermatitis or psoriasis.   A process for the preparation of 4-methylsulfonyl-2-butenenitrile which comprises reacting sodium methanesulphinate with 4-bromo-2-butenenitrile. Bromine Br ₂ is reacted with allyl cyanide, and by adding a basic solution further comprises the preparation of 4-bromo-2-butenenitrile, The method of claim 13.