CN106344595B

CN106344595B - Application of algin oligosaccharide and derivatives thereof in preparation of pain treatment drugs

Info

Publication number: CN106344595B
Application number: CN201510424357.7A
Authority: CN
Inventors: 耿美玉; 辛现良; 丁健
Original assignee: Shanghai Green Valley Pharmaceutical Co Ltd
Current assignee: Shanghai Green Valley Pharmaceutical Co Ltd
Priority date: 2015-07-17
Filing date: 2015-07-17
Publication date: 2020-06-19
Anticipated expiration: 2035-07-17
Also published as: CN106344595A

Abstract

The invention relates to algin oligosaccharide and derivatives thereof, in particular to mannuronic acid oligosaccharide formed by β -D-mannuronic acid through 1, 4 glycosidic bond connection or mannuronic acid oligosaccharide with carboxyl at the 1-position of the reducing end, derivatives thereof or pharmaceutically acceptable salts thereof.

Description

Application of algin oligosaccharide and derivatives thereof in preparation of pain treatment drugs

Technical Field

The invention relates to algin oligose and its derivative and their application in treating pain.

Background

Pain (pain) is a complex physiological and psychological activity and is one of the most common symptoms in the clinic. Acute pain is often a symptom of other diseases, while chronic pain is itself a disease. At present, about 30% of people all over the world suffer from chronic pain, and at least more than 1 hundred million pain patients in China. Migraine is one of the clinically common types of headache. It is characterized by recurrent headache, and the intermittent period of attack is normal. The exact cause of migraine is not clear, and clinically, symptomatic treatment and preventive treatment during the attack are the main causes, and no ideal therapeutic medicine exists.

Algin is a main component of the cell wall of brown algae, and is linear anionic polysaccharide formed by connecting β -D-mannuronic acid and α -L-guluronic acid through 1, 4-glycosidic bond, is a high molecular compound with large molecular weight generally between tens of thousands to millions of daltons, has rich sources, and is widely applied to food, chemical and pharmaceutical industries and the like.

However, the molecular weight of algin is large, so that the algin is limited in the aspect of drug application. Therefore, the alginate oligosaccharides prepared by various degradation methods, including mannuronic acid oligosaccharide, guluronic acid oligosaccharide, glycuronic acid oligosaccharide and derivatives thereof, have important research values in the research fields of glycochemistry, glycobiology, glycoengineering, carbohydrate drugs and the like. Many methods can be used to degrade algin to obtain alginate oligosaccharides, including enzymatic, chemical and physical degradation methods.

Disclosure of Invention

The invention provides alginate oligosaccharides and derivatives thereof and use thereof in treating pain, the alginate oligosaccharides and derivatives thereof are alginate hydrolysis degradation products, preferably, the alginate oligosaccharides and derivatives thereof with molecular weight in the range of 300-4500Da or pharmaceutically acceptable salts of the oligosaccharides and derivatives thereof, the alginate oligosaccharides and derivatives thereof or pharmaceutically acceptable salts of the oligosaccharides and derivatives thereof are mannuronic acid oligosaccharides formed by β -D-mannuronic acid through 1, 4 glycosidic bond connection or mannuronic acid oligosaccharides with carboxyl at the 1-reducing end and derivatives thereof, or pharmaceutically acceptable salts thereof, preferably, the invention also relates to alginate oligosaccharides and derivatives thereof of formula (I) or formula (II) or pharmaceutically acceptable salts of the oligosaccharides and derivatives thereof.

The present invention relates, in one aspect, to an alginate oligosaccharide represented by the following structural formula (I) and derivatives thereof, or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof,

in formula (I), n represents one or more integers from 0 to 20, for example n is one or more integers selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20.

The invention also relates to the algin oligosaccharide and the derivative thereof represented by the following structural formula (II), or the pharmaceutically acceptable salt of the oligosaccharide and the derivative thereof,

in formula (II), n represents one or more integers from 0 to 20, for example n is one or more integers selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20.

In the above formulae (I) and (II), n is preferably 0 to 10, more preferably 2 to 8, and most preferably 4. The reason why alginate oligosaccharides with n-0 to 10 (preferably n-2 to 8, most preferably n-4) have better biological effects is not clear, and it is likely that they are easily recognized and accepted by body cells. In some embodiments, n may also be one or more integers selected from the range of 0 to 20.

The algin oligosaccharide of formula (I) and (II) of the invention can be prepared and identified by the method of patent number ZL 200580009396.5. In a preferred embodiment of the present invention, the alginate oligosaccharide represented by structural formula (I) is prepared by the following steps: dissolving a solution containing polymannuronate sodium salt, which is a component of algin and is commercially available or prepared, for example, with reference to Chinese patent ZL200580009396.5The liquid (preferably aqueous solution) is placed in an autoclave and reacted for 2 to 6 hours under the conditions of pH2 to 6 and temperature 100 and 120 ℃. The reacted solution is then removed from the autoclave and neutralized (e.g., with NaOH solution) to neutrality. The neutral solution is slowly added to ethanol (e.g., industrial ethanol, 95% ethanol, or absolute ethanol) with stirring, and subjected to alcohol precipitation. Then, the solid substance obtained by the alcohol precipitation is separated by filtration (for example, suction filtration), and the solid substance obtained by the filtration is preferably washed with ethanol (preferably, absolute ethanol) during the filtration. Drying the solid matter (usually white) to obtain crude mixture of alginate oligosaccharide represented by formula (I). Preferably, the crude alginate-derived oligosaccharide mixture can be further prepared into a solution (preferably an aqueous solution), and then 95% ethanol is added into the solution for alcohol precipitation again. The precipitate obtained is separated by filtration and again precipitated with ethanol and optionally washed with absolute ethanol. The precipitate was separated and dried to give a solid material. The solid material is brought into solution, preferably an aqueous solution, the solution is filtered through a filter membrane, for example a filter membrane with a pore size of 3 μm, and the filtrate is collected. Eluting the filtrate with size exclusion chromatography (Bio-Gel-P6 or Bio-Gel-P10 Gel column, e.g. 1.6X 180cm or other size) to obtain mobile phase as eluent NH₄HCO₃And so on. The eluates were collected from the chromatogram in sequence using a plurality of vessels, and the sugar content in the eluates of each vessel was detected by a sulfuric acid-carbazole method. The sulfuric acid-carbazole method detection method is to use sulfuric acid-carbazole to develop color, detect with a conventional ultraviolet spectrometer, and express the result as an optical density value (OD value) and express that different optical density values show that the sugar content is different. For convenience, the different molecular weight alginate oligosaccharides may be referred to as

components

1, 2, 3. And respectively collecting eluates containing alginate oligosaccharide components with different molecular weights according to the detection result. The eluates containing different molecular weight alginate oligosaccharide components (such as

components

1, 2, 3.. times.etc.) are each separately concentrated, desalted, and freeze-dried to obtain alginate oligosaccharides represented by formula (I) having different molecular weights. The alginate oligosaccharides with different molecular weights have different values of n corresponding to the alginate oligosaccharides shown in the formula (I). When the target product is alginate-oligosaccharide mixtureWhen the target mixture is obtained, the eluents containing different alginate oligosaccharide components are respectively dried and then mixed, or the eluents containing different alginate oligosaccharide components are combined and then dried, so that the target mixture is obtained.

The preparation method of the algin oligosaccharide shown in the structural formula (II) comprises the following steps: and (3) reacting the crude alginate oligosaccharide mixture of the formula (I) which is not subjected to size exclusion chromatography separation or the separated alginate oligosaccharides of the formula (I) with different n values with an oxidizing agent under the heating condition to obtain the alginate oligosaccharides of the formula (II). In one embodiment, the oxidizing agent is copper hydroxide (e.g., obtained by adding a copper sulfate solution to a sodium hydroxide solution and immediately mixing). Adding fresh oxidant into the crude alginate-derived oligosaccharide mixture of formula (I) which is not subjected to size exclusion chromatography or the solution (such as aqueous solution) prepared from the separated alginate-derived oligosaccharides of formula (I) with different n values, and heating for reaction until no brick red precipitate is generated. The reaction system was subjected to centrifugation to remove the precipitate to obtain a supernatant. For the purpose of checking whether the oxidation reaction is complete, a small amount of supernatant may be taken to add the oxidizing agent again, and it is checked whether a brick-red precipitate is still generated. If brick red precipitate is generated, the whole supernatant obtained by the centrifugation and another part of the oxidant are continuously reacted until no brick red precipitate is generated during the inspection. The resulting reaction system was centrifuged to obtain a supernatant. Ethanol (e.g., industrial ethanol, 95% ethanol, or absolute ethanol) is added to the resulting supernatant to carry out alcohol precipitation. Filtering and separating solid substance obtained by alcohol precipitation, and washing the solid substance with absolute ethyl alcohol. The resulting solid material was dried. The separation was performed according to the same size exclusion chromatography separation method as the preparation method of the alginate oligosaccharides of formula (I) above. Thereby obtaining the algin oligose shown in the formula (II) with different molecular weights. The alginate oligosaccharides with different molecular weights have different values of n corresponding to the alginate oligosaccharides represented by the formula (II). When the target product is the alginate-derived oligosaccharide mixture, the target mixture can be obtained by drying the eluents containing different alginate-derived oligosaccharide components respectively and then mixing, or by combining the eluents containing different alginate-derived oligosaccharide components and then drying.

The derivative of the algin oligosaccharide or the pharmaceutically acceptable salt of the formula (I) or the formula (II) comprises ester of which one or more hydroxyl groups are esterified by inorganic acid or organic acid. The organic acid capable of forming an ester with one or more hydroxyl groups of the alginate oligosaccharide of formula (I) or formula (II) of the present invention or a pharmaceutically acceptable salt thereof includes, but is not limited to: formic acid, acetic acid, oxalic acid, glycolic acid, propionic acid, malonic acid, butyric acid, succinic acid, valeric acid, acrylic acid, oxalic acid, maleic acid, fumaric acid, malic acid, succinic acid, citric acid, tartaric acid, lactic acid, methanesulfonic acid, lactic acid, salicylic acid, acetylsalicylic acid, benzenesulfonic acid, p-toluenesulfonic acid, pyruvic acid, hydroxybutyric acid, adipic acid, phthalic acid, mandelic acid, benzoic acid, boric acid, and the like. The inorganic acid capable of forming an ester with one or more hydroxyl groups of the alginate oligosaccharide of formula (I) or formula (II) of the present invention or a pharmaceutically acceptable salt thereof includes, but is not limited to: sulfuric acid, sulfurous acid, phosphoric acid, metaphosphoric acid, phosphorous acid, hypophosphorous acid, pyrophosphoric acid, polyphosphoric acid, and the like.

The pharmaceutically acceptable salts of the oligosaccharide of formula (I) or formula (II) and derivatives thereof of the present invention comprise: inorganic salts such as lithium salt, sodium salt, potassium salt, beryllium salt, magnesium salt, calcium salt, iron salt, zinc salt, selenium salt, vanadium salt, tin salt, silicon salt, strontium salt, or basic addition salts with a basic amino acid such as lysine, arginine, ornithine, etc., among which sodium salt is preferred. The pharmaceutically acceptable salts can be prepared by conventional methods.

The medicine for treating pain comprises the algin oligosaccharide shown in the formula (I) or the formula (II) and a derivative thereof, or a pharmaceutically acceptable salt of the oligosaccharide and the derivative thereof, and one or more pharmaceutically acceptable carriers. The drug of the present invention may be in the form of tablets, hard capsules, soft capsules, enteric capsules, microcapsules, granules, syrups, injections, granules, emulsions, suspensions, solutions, and sustained-release preparations for oral or non-oral administration.

The pharmaceutically acceptable carrier of the present invention refers to pharmaceutically acceptable carriers well known to those skilled in the art, and the pharmaceutically acceptable carriers of the present invention include, but are not limited to: fillers, wetting agents, binders, disintegrants, lubricants, binders, glidants, taste masking agents, surfactants, preservatives, and the like. Fillers include, but are not limited to, lactose, microcrystalline cellulose, starch, powdered sugar, dextrin, mannitol, calcium sulfate, and the like. Wetting agents and binders include, but are not limited to, sodium carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, gelatin, sucrose, polyvinylpyrrolidone, and the like. Disintegrants include, but are not limited to, sodium carboxymethyl starch, crospovidone, croscarmellose sodium, low substituted hydroxypropyl cellulose, and the like. Lubricants include, but are not limited to, magnesium stearate, aerosil, talc, hydrogenated vegetable oils, polyethylene glycols, magnesium lauryl sulfate, and the like. Binders include, but are not limited to, acacia, alginic acid, carboxymethylcellulose calcium, carboxymethylcellulose sodium, dextrates, dextrin, dextrose, ethylcellulose, gelatin, liquid glucose, guar gum, hydroxyethyl cellulose, hydroxypropyl methylcellulose, magnesium aluminum silicate, maltodextrin, methylcellulose, polymethacrylates, polyvinylpyrrolidone, pregelatinized starch, sodium alginate, sorbitol, starch, syrup, and tragacanth. Glidants include, but are not limited to, colloidal silicon dioxide, powdered cellulose, magnesium trisilicate, silicon dioxide, and talc. Taste-masking agents include, but are not limited to, aspartame, stevioside, fructose, glucose, syrup, honey, xylitol, mannitol, lactose, sorbitol, maltitol, glycyrrhizin. Surfactants include, but are not limited to, tween-80, poloxamers. Preservatives include, but are not limited to, parabens, sodium benzoate, potassium sorbate, and the like.

Methods of preparing various pharmaceutical compositions containing various proportions of active ingredients are known or will be apparent to those skilled in the art in light of the present disclosure. Such as REMINGTON' S PHARMACEUTICAL SCIENCES, Martin, E.W., ed., Mack Publishing Company, 19th ed. (1995). The process for preparing the pharmaceutical composition comprises incorporating suitable pharmaceutical excipients, carriers, diluents and the like.

The pharmaceutical formulations of the present invention are manufactured in a known manner, including conventional mixing, dissolving or lyophilizing processes.

The medicaments of the invention are administered in various routes suitable for the selected mode of administration, for example orally or parenterally (by intravenous, intramuscular, topical or subcutaneous routes).

Thus, the agents of the invention may be administered systemically, e.g., orally, in a suitable pharmaceutically acceptable carrier, such as an inert diluent or an edible carrier. They may be enclosed in hard or soft shell gelatin capsules and may be compressed into tablets. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of swallowable tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such formulations should contain at least 0.1% active compound. The proportion of active compound in such formulations may, of course, vary and may range from about 0.01% to about 99% by weight of a given unit dosage form. In such therapeutically useful pharmaceutical formulations, the amount of active compound is such that an effective dosage level is obtained.

Tablets, troches, pills, capsules and the like may also contain: binders, such as gum tragacanth, acacia, corn starch or gelatin; excipients, such as dicalcium phosphate; disintegrating agents, such as corn starch, potato starch, alginic acid, and the like; lubricants, such as magnesium stearate; and sweeteners such as sucrose, fructose, lactose or aspartame; or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavor. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a vegetable oil or polyethylene glycol. Various other materials may be present, as coatings, or to otherwise modify the physical form of the solid unit dosage form. For example, tablets, pills, or capsules may be coated with gelatin, wax, shellac, or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl or propylparabens as preservatives, a dye and a flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and non-toxic in the amounts employed. In addition, the active compounds may be incorporated into sustained release formulations and sustained release devices.

The active compounds may also be administered intravenously or intraperitoneally by infusion or injection. Aqueous solutions of the active compounds or their salts, optionally in admixture with non-toxic surfactants, may be prepared. Dispersants in glycerol, liquid polyethylene glycols, triacetin and mixtures thereof, and oils may also be prepared. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical dosage forms suitable for injection or infusion may include sterile aqueous solutions or dispersions or sterile powders of the active ingredient, optionally encapsulated in liposomes, containing ready-to-use preparations of injectable or infusible solutions or dispersions suitable for sterility. In all cases, the final dosage form must be sterile, liquid and stable under the conditions of manufacture and storage. The liquid carrier can be a solvent or liquid dispersion medium including, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. Suitable fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersants, or by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use of compositions which delay absorption of the agent (e.g., aluminum monostearate and gelatin).

Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze-drying technique which yield a powder of the active ingredient plus any additional required ingredients present in the sterile-filtered solution.

Useful solid carriers include finely divided solids (e.g., talc, clay, microcrystalline cellulose, silicon dioxide, alumina, and the like). Useful liquid carriers include water, ethanol or ethylene glycol or water-ethanol/ethylene glycol mixtures in which the combination of the invention may be dissolved or dispersed in effective amounts, optionally with the aid of non-toxic surfactants. Adjuvants (such as fragrances) and additional antimicrobial agents may be added to optimize the properties for a given use.

Thickeners (e.g., synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified inorganic materials) can also be used with liquid carriers to form coatable pastes, gels, ointments, soaps, etc., for direct application to the skin of the user.

The therapeutically required amount of the compound or its active salt or derivative will depend not only on the particular salt selected, but also on the mode of administration, the nature of the disease to be treated and the age and condition of the patient, and will ultimately be at the discretion of the attendant physician or clinician.

The formulations may be presented in unit dosage form comprising physically discrete units of a unit dose suitable for administration to the human or other mammalian body. The unit dosage form may be a capsule or tablet, or a plurality of capsules or tablets. The amount of unit dose of the active ingredient may be varied or adjusted from about 0.01 to about 1000 mg or more depending upon the particular treatment involved.

In some embodiments, the present invention also relates to alginate oligosaccharides and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharides and derivatives thereof for use in the treatment of pain, said alginate oligosaccharides and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharides and derivatives thereof for use in the treatment of pain are selected from the group consisting of mannuronic acid oligosaccharides formed by β -D-mannuronic acid through a 1, 4 glycosidic linkage or mannuronic acid oligosaccharides having a carboxyl group at the 1-position of the reducing end thereof and derivatives thereof, or pharmaceutically acceptable salts thereof in some preferred aspects, said alginate oligosaccharides and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharides and derivatives thereof for use in the treatment of pain are selected from the group consisting of alginate oligosaccharides and derivatives thereof represented by the following structural formula (I) or formula (II), or pharmaceutically acceptable salts of said oligosaccharides and derivatives thereof,

wherein n in each of formula (I) or formula (II) is one or more integers selected from 0 to 20. The alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of the oligosaccharide and derivatives thereof for treating pain may also be selected from alginate oligosaccharides represented by formula (I) or formula (II) and derivatives thereof, or pharmaceutically acceptable salts of the oligosaccharides and derivatives thereof, and n is one or more integers selected from 0 to 10. The alginate oligosaccharide and derivatives thereof or the pharmaceutically acceptable salts of the oligosaccharide and derivatives thereof for treating pain may also be selected from alginate oligosaccharides represented by formula (I) or formula (II) and derivatives thereof, or pharmaceutically acceptable salts of the oligosaccharides and derivatives thereof, and n is one or more integers selected from 2 to 8. The alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of the oligosaccharide and derivatives thereof for use in the treatment of pain may also be selected from alginate oligosaccharides and derivatives thereof represented by formula (I) or formula (II) or pharmaceutically acceptable salts of the oligosaccharides and derivatives thereof, and n is 4.

In some embodiments, the present invention also relates to a method for treating pain, comprising administering to a patient in need thereof a therapeutically effective amount of an alginate oligosaccharide and derivatives thereof or a pharmaceutically acceptable salt of the oligosaccharide and derivatives thereof according to the present invention, in some embodiments, the method for treating pain comprises administering to a patient in need thereof a therapeutically effective amount of an alginate oligosaccharide and derivatives thereof or a pharmaceutically acceptable salt of the oligosaccharide and derivatives thereof according to the present invention, wherein the alginate oligosaccharide and derivatives thereof or the pharmaceutically acceptable salt of the oligosaccharide and derivatives thereof is selected from the group consisting of a mannuronic acid oligosaccharide formed by 1, 4 glycosidic linkage of β -D-mannuronic acid or a mannuronic acid oligosaccharide having a carboxyl group at the 1-position of the reducing end thereof and derivatives thereof, or a pharmaceutically acceptable salt thereof, in some embodiments, the method for treating pain comprises administering to a patient in need thereof a therapeutically effective amount of an alginate oligosaccharide and derivatives thereof or a pharmaceutically acceptable salt of the oligosaccharide and derivatives thereof represented by the following structural formula (I) or formula (II) or a pharmaceutically acceptable salt of the alginate oligosaccharide and derivatives thereof,

wherein n in each of formula (I) or formula (II) is one or more integers selected from 0 to 20. In some embodiments, the method for treating pain comprises administering to a patient in need of treatment a therapeutically effective amount of an alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof according to the invention, wherein the alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof are selected from alginate oligosaccharides and derivatives thereof represented by formula (I) or formula (II) or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof, and n is one or more integers selected from 0 to 10. In some embodiments, the method for treating pain comprises administering to a patient in need of treatment a therapeutically effective amount of an alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof according to the invention, wherein the alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof are selected from alginate oligosaccharides and derivatives thereof represented by formula (I) or formula (II) or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof, and n is one or more integers selected from 2 to 8. In some embodiments, the method for treating pain comprises administering to a patient in need of treatment a therapeutically effective amount of an alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof according to the invention, wherein the alginate oligosaccharide and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof are selected from the group consisting of alginate oligosaccharides and derivatives thereof represented by formula (I) or formula (II) or pharmaceutically acceptable salts of said oligosaccharide and derivatives thereof, and n is 4.

The term "treating" as used herein generally refers to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic, in terms of preventing the disease or its symptoms, in whole or in part; and/or may be therapeutic in terms of partially or completely stabilizing or curing the disease and/or side effects due to the disease. As used herein, "treatment" encompasses any treatment of a disease in a patient, including: (a) preventing a disease or condition in a patient susceptible to the disease or condition but not yet diagnosed as having the disease; (b) inhibiting the symptoms of the disease, i.e., arresting its development; or (c) alleviating the symptoms of the disease, i.e., causing regression of the disease or symptoms.

In all embodiments of the present invention, the alginate oligosaccharides and derivatives thereof according to the present invention further comprise a mixture comprising said one or more alginate oligosaccharides and derivatives thereof or pharmaceutically acceptable salts of said oligosaccharides and derivatives thereof. For example, n may also be a plurality of integers selected from each of the numerical ranges recited.

On the other hand, in all embodiments of the present invention, the numerical ranges of the present invention are intended to cover any integer within the ranges. For example, in all embodiments of the present invention, n in the alginate oligosaccharide represented by formula (I) or formula (II) and derivatives thereof or pharmaceutically acceptable salts of the oligosaccharide and derivatives thereof may be any integer in the range of 0 to 20, such as n may be one or more integers selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20. Unless otherwise specified, percentages recited herein refer to weight percentages.

In all embodiments of the invention, the pain of the invention includes, but is not limited to, the types of pain listed in the examples, for example, acute and chronic pain such as neuropathic and post-operative pain, chronic lower back pain, cluster headache, herpetic neuralgia, phantom limb pain, central pain, dental pain, opioid-resistant pain, visceral pain, surgical pain, bone injury pain, pain during exertion and childbirth, pain due to burns including sunburn, postpartum pain, migraine, angina, and pain associated with the genitourinary tract (including cystitis), vascular pain, neuropathic pain, migraine, trigeminal neuralgia, intercostal neuralgia, surgical incision pain, chronic fasciitis pain, heel pain, diabetic peripheral neuropathy pain, phantom limb pain, muscle pain, bone pain, joint pain, cancerous pain, noncancerous pain, and the like. The term also includes nociceptive pain or nociception.

The invention will now be illustrated by way of examples. Those skilled in the art will appreciate that these embodiments are illustrative, not restrictive. These examples are not intended to limit the scope of the present invention in any way.

Drawings

FIG. 1: an alginate oligosaccharide column separation diagram shown in a formula (I). A: p6 column isolation diagram; b: p10 column isolation diagram.

FIG. 2: an alginate oligosaccharide column separation diagram shown in a formula (II). A: p6 column isolation diagram; b: p10 column isolation diagram.

Detailed Description

EXAMPLE one preparation of alginate oligosaccharides

1g of sodium polymannuronate (weight-average molecular weight 8235Da, supplied by Shanghai green valley pharmaceutical Co., Ltd.) was weighed and added with an appropriate amount of distilled water to prepare a 1% (by weight) aqueous solution of sodium polymannuronate. The pH of the 1% aqueous solution of sodium polymannuronate was adjusted to 4 with hydrochloric acid, and then the aqueous solution was placed in an autoclave. The reaction was heated at 110 ℃ for 4 hours. The reacted solution was taken out of the autoclave and allowed to cool. After cooling, the pH value of the solution after the reaction was adjusted with NaOH solution to give a neutral liquid. Slowly adding the neutral liquid into ethanol with the volume of 4 times that of the liquid under the stirring condition, carrying out alcohol precipitation and standing overnight. Filtering and separating solid substances obtained by alcohol precipitation, washing the solid substances obtained by filtering and separating with absolute ethyl alcohol during filtering and separating, and finally obtaining white filter cakes. And (3) drying the filter cake in a 60 ℃ oven to obtain the crude product of the algin oligosaccharide.

Taking 1g of the crude product of the algin oligosaccharide, preparing a 10% aqueous solution, and carrying out alcohol precipitation again by using a 95% ethanol solution. Filtering, separating, precipitating again with ethanol, and washing the precipitate with anhydrous ethanol. Separating the precipitateAnd dried to obtain a solid substance. The solid material was prepared as a 5% (by weight) aqueous solution, which was filtered through a 3 μm pore membrane and the filtrate was collected. The filtrate was eluted on a column (1.6X 180cm, available from Bio-Rad) of a BioGel-P6 Gel (molecular exclusion chromatography) to give a mobile phase containing 0.2 mol. L of eluent^-1NH₄HCO₃. The eluate was collected from the column chromatography using a plurality of 5ml test tubes in sequence, and then the sugar content of the eluate in each of the test tubes was detected by a sulfuric acid-carbazole method. And respectively collecting eluates containing alginate oligosaccharide components with different molecular weights according to the detection result. The eluates containing different molecular weight alginate-derived oligosaccharide fractions were each separately concentrated under reduced pressure and freeze-dried, and fraction 1 was discarded to obtain alginate-derived oligosaccharide fractions 2-12 (n has a value of 0-10, respectively, as can be seen from subsequent detection) represented by formula (I) each having a different molecular weight (Volume in the figure represents the Volume of the eluate in terms of the number of test tubes) (FIG. 1A). The fractions difficult to be separated from the Bio-Gel-P6 Gel column were further eluted and separated by a Bio-Gel-P10 Gel column (1.6X 180cm, available from Bio-Rad) and the eluent used as the mobile phase was 0.2 mol. L^- ¹NH₄HCO₃. The eluates were collected in a plurality of 5ml test tubes in sequence, and then the sugar content of the eluates in each test tube was measured by a sulfuric acid-carbazole method. And respectively collecting eluates containing alginate oligosaccharide components with different molecular weights according to the detection result. The eluates containing the alginate oligosaccharide fractions of different molecular weights were each separately concentrated under reduced pressure and freeze-dried to obtain alginate oligosaccharide fractions 13-24 (n has a value of 11-22, respectively, according to the subsequent examination) of formula (I) having different molecular weights (Volume in the figure represents the Volume of the eluates in terms of the number of test tubes) (FIG. 1B). The Bio-Gel-P6 column and the P10 column are two polyacrylamide microspheres with different molecular size exclusion, the algin oligosaccharide with the molecular weight below 3000Dalton can be better separated on the P6 column, and the algin oligosaccharide with the molecular weight below 3000-6000Dalton can be better separated on the P10 column. Through ultraviolet spectrum analysis, infrared spectrum analysis and comparison with available reference,¹H-NMR spectroscopic analysis,¹³After C-NMR spectroscopic analysis and mass spectrometric analysis, it was found that PIn the 6 column diagram (FIG. 1A) and the P10 column diagram (FIG. 1B), alginate oligosaccharide compounds represented by formula (I) are obtained, wherein n is 0-22, and the fractions 2-24 are respectively. According to the spectrogram result, the algin oligosaccharide eluent with the n-2-8 formula (I) is combined and dried, and then the algin oligosaccharide mixture with the n-2-8 formula (I) is obtained.

EXAMPLE preparation of algin oligosaccharide oxo-decomposition product

5g of the crude product of the algin oligosaccharide which is not separated by the molecular exclusion chromatography is prepared into 5 percent (weight percentage) of aqueous solution. The fresh oxidant copper hydroxide was prepared by adding 25ml of 5% by weight copper sulfate solution to 50ml of 10% by weight sodium hydroxide solution and immediately mixing. The fresh oxidant copper hydroxide was immediately added to 40ml of the above 5% (by weight) alginate oligosaccharide solution while heating in a boiling water bath until no more brick-red precipitate was produced. The reaction system was subjected to centrifugation to remove the precipitate to obtain a supernatant. A little supernatant was taken and added again to the oxidant to check if any brick red precipitate was produced. If brick-red precipitate is generated, the whole supernatant obtained by the centrifugation is continuously reacted with another part of the oxidant until no brick-red precipitate is generated. The resulting reaction system was centrifuged to obtain a supernatant. Adding 95% ethanol with 4 times volume amount to the supernatant, precipitating with ethanol, and standing overnight. Filtering and separating solid substance obtained by alcohol precipitation, and washing the solid substance with absolute ethyl alcohol. And (3) drying the obtained solid substance in a drying oven at 60 ℃ to obtain the crude product of the algin oligosaccharide shown in the formula (II).

Taking 1g of the crude product of the algin oligosaccharide to prepare 10 percent (weight percentage) of water solution, carrying out alcohol precipitation again by using 95 percent of ethanol solution, filtering, separating and carrying out alcohol precipitation again to obtain precipitate, and optionally washing by using absolute ethanol. The precipitate was separated and dried to give a solid material. The solid material was prepared as a 5% (by weight) aqueous solution, which was filtered through a 3 μm pore membrane and the filtrate was collected. The filtrate was eluted on a column (1.6X 180cm, available from Bio-Rad) of a Biogel 6 Gel (size exclusion chromatography: BioGel-P6) to give a mobile phase of 0.2mo as eluentl·L^-1NH₄HCO₃. The eluate was collected from the column chromatography using a plurality of 5ml test tubes in sequence, and then the sugar content of the eluate in each of the liquid collection tubes was measured by a sulfuric acid-carbazole method. And respectively collecting eluates containing alginate oligosaccharide components with different molecular weights according to the detection result. The eluates containing different molecular weight alginate-derived oligosaccharide fractions were each separately concentrated under reduced pressure and freeze-dried, and fraction 1 was discarded to obtain alginate-derived oligosaccharide fractions 2-12 (n has a value of 0-10, respectively, as can be seen from subsequent detection) represented by formula (II) each having a different molecular weight (Volume in the figure represents the Volume of the eluate in terms of the number of test tubes) (FIG. 2A). The fractions difficult to be separated from the Bio-Gel-P6 Gel column were further eluted and separated by a Bio-Gel-P10 Gel column (1.6X 180cm, available from Bio-Rad) and the eluent used as the mobile phase was 0.2 mol. L^-1NH₄HCO₃. The eluates were collected in a plurality of 5ml test tubes in sequence, and then the sugar content of the eluates in each test tube was measured by a sulfuric acid-carbazole method. And respectively collecting eluates containing alginate oligosaccharide components with different molecular weights according to the detection result. The eluates containing the alginate oligosaccharide fractions of different molecular weights were each separately concentrated under reduced pressure and freeze-dried to obtain alginate oligosaccharide fractions 13-23 (n has a value of 11-21, respectively, as can be seen from subsequent detection) of formula (II) having different molecular weights (Volume in the figure represents the Volume of the eluates in terms of the number of test tubes) (FIG. 2B). The Bio-Gel-P6 column and the P10 column are two polyacrylamide microspheres with different molecular size exclusion, the algin oligosaccharide with the molecular weight below 3000Dalton can be better separated on the P6 column, and the algin oligosaccharide with the molecular weight below 3000-6000Dalton can be better separated on the P10 column. Through ultraviolet spectrum analysis, infrared spectrum analysis and comparison with available reference,¹H-NMR spectroscopic analysis,¹³After C-NMR spectroscopic analysis and mass spectrometric analysis, it was found that the alginate oligosaccharide compounds of the formula (II) wherein n is 0 to 21, of fractions 2 to 23 were obtained in the P6 column (FIG. 2A) and the P10 column (FIG. 2B), respectively. According to the spectrogram result, collecting and combining the algin oligosaccharide eluent represented by the formula (II) with n being 2-8, drying the algin oligosaccharide eluent, and then obtaining the algin oligosaccharide mixture represented by the formula (II) with n being 2-8.

Example Effect of Tri-alginate oligosaccharide derivatives on acetic acid-induced pain in mice

1 materials of the experiment

1.1 drugs and reagents

The preparation method of the second example is adopted to prepare the algin oligosaccharide (n is 2-8, namely the components 4-10 in the figure) mixture of the formula (II). The mixture is white powder, and is easily soluble in water; acetic acid, manufactured by Kaiton chemical reagents, Inc. of Tianjin; aspirin, produced by Shijiazhuang Europe pharmaceuticals Co., Ltd.

1.2 Experimental animals

Kunming mouse, male and female half, weight 18-22g, by Shanghai Xipul-Bikai laboratory animals company Limited.

2 method of experiment

2.1 animal grouping and administration and treatment

Mice were randomly divided into 5 groups, namely blank control group, positive drug aspirin 200mg/kg group, alginate oligosaccharide derivatives 25mg/kg, 50mg/kg, 100mg/kg group, each group containing 10 animals. From the grouping day, the blank control group was gavaged with 20ml/kg distilled water per day, and the other groups were gavaged with the corresponding drugs 1 time per day for 7 days. 1 hour after the last administration, each group of mice was intraperitoneally injected with 0.6% acetic acid solution (0.2 ml), and the writhing latency (i.e., the time from the acetic acid injection to the occurrence of writhing reaction) and the number of writhing times of the mice within 20 minutes after the acetic acid injection were recorded, and the writhing inhibition rate and the analgesic percentage (%) were calculated according to the following formulas.

The writhing inhibition ratio (%) was (average writhing number in blank control group-average writhing number in administration group)/average writhing number in blank control group × 100%

Percent (%) pain (number of animals without writhing in experimental group-number of animals without writhing in blank control group)/number of animals in experimental group ]. times.100%

3 statistical treatment

The results were statistically analyzed and expressed as "mean ± standard deviation" (mean ± SD) and compared using analysis of variance (ANOVA). The difference is significant when p is less than 0.05, and the difference is very significant when p is less than 0.01.

4 results of the experiment

4.1 Effect of alginate-derived oligosaccharide derivatives on acetic acid-induced writhing response in mice

The injection of chemical substances such as acetic acid solution into abdominal cavity of mouse can stimulate peritoneum of mouse to cause intermittent persistent pain, which is manifested by abdominal indent, abdominal anterior wall close to cage bottom, hip distortion and hind limb extension, and a special posture called writhing reaction. Writhing latency (i.e., the time from acetic acid injection to writhing response) and the number of writhing times over a period of time may represent the severity of the pain.

The results of the experiments on the writhing reaction of the alginate-derived oligosaccharide to the acetic acid in mice are shown in Table 1. The algin oligosaccharide derivative has good inhibition effect on mouse writhing reaction, and the dosage groups present dosage dependence relationship, and the high dosage group has the best effect. Compared with the blank control group, the differences of the number of writhing times, the inhibition rate of writhing and the percentage of analgesia have statistical significance (P is less than 0.01). The alginate-derived oligosaccharide derivative has pain relieving effect.

TABLE 1 Effect of alginate oligosaccharide derivatives on acetic acid-induced writhing response in mice (n 10, mean + -SD)

Blank control group with p < 0.05 p < 0.01 vs

Similar results were also obtained when the above experiments were performed with alginate oligosaccharides of formula (I) or formula (II), respectively, each n being a single integer selected from 0 to 20, or a pharmaceutically acceptable salt thereof.

Example Effect of Tetraalginate oligosaccharide derivatives on nitroglycerin-induced migraine in rats

1 materials of the experiment

1.1 drugs and reagents

The preparation method of the second example is adopted to obtain the algin oligosaccharide (n ═ 2-8, namely the component 4-1O in the figure) mixture of the formula (II). The mixture is white-like powder, and is easily soluble in water.

1.2 Experimental animals

SD male rats weighing 180-.

2 method of experiment

2.1 animal grouping and administration

SD rats are randomly divided into a blank control group, a model group and alginate oligosaccharide derivatives 12.5, 25, 50, 100 and 200mg/kg groups, and each group comprises 8 rats. The administration is started on the grouping day, distilled water is given to the blank control group and the model group by intragastric administration, the corresponding drugs are given to the other groups, 1 time is given each day, the administration is continuously carried out for 28 days, 30 minutes is carried out after the last administration, and the other groups except the blank control group are injected with nitroglycerin 10mg/kg subcutaneously at the right shoulder for molding. Observing the appearance time and duration of ear red after the rat model is made, and the times of head bending in two time periods of 30-45 minutes and 60-75 minutes after the model is made; the 5-HT content of brain tissue is determined by fluorescence spectrophotometry. Measured at a wavelength of Ex356nm/Em 483nm, and the results are expressed in ng/g brain weight.

3 statistical treatment

4 results of the experiment

Migraine is a functional disorder of blood vessels and nerves under the interaction of blood vessels and nerve mechanisms. Nitroglycerin can cause hypersensitivity of trigeminal nerve fiber and cause migraine by dilating blood vessels of meninges, forming neurogenic inflammation, activating functions of neurons of hypothalamus, brain stem and spinal cord segments and the like. The nitroglycerin model is an animal model established in 1995 and is a classical animal migraine model at present. According to the nitroglycerin pathogenesis, the ear red time of a model animal caused by vasodilatation, the number of times of head scratching caused by pain and the content of 5-hydroxytryptamine (5-HT) which is a pain sensitive factor caused by brain tissues are detected, so that the severity of the migraine can be evaluated.

The results of this study show (Table 2-4) that about 3 minutes after the subcutaneous injection of nitroglycerin, rats show red ears which last for about 2.5 hours; and the number of times of head bending in the 30-45 min and 60-75 min periods after molding is obviously more than that of the blank control group, and the 5-HT content of the brain tissue is obviously lower than that of the blank control group. Under the dosage of 25-200mg/kg, the algin oligosaccharide derivative can obviously delay the appearance time of rat ear red, shorten the duration time of the ear red, reduce the times of head bending in two time periods of 30-45 minutes and 60-75 minutes and increase the content of 5-HT in brain tissues. The alginate oligosaccharide derivative can obviously inhibit rat migraine, and has the function of treating pain.

TABLE 2 Effect of alginate oligosaccharide derivatives on the appearance and duration of nitroglycerin migraine headache in rat ear Red (n 8, mean + -SD)

Blank control group with # p less than 0.01 vs; p < 0.05, p < 0.01 vs model set

TABLE 3 Effect of alginate oligosaccharide derivatives on nitroglycerin migraine headache-induced rat head-bending times (n ═ 8, mean. + -. SD)

Blank control group with # p less than 0.01 vs; p < 0.05, p < 0.01 vs model set

TABLE 4 Effect of alginate oligosaccharide derivatives on 5-HT content in Nitropyrlycerol migraine-induced rat brain tissue (n ═ 8, mean. + -. SD)

# p < 0.01 vs to white control; model set of p < 0.01 vs

Example Effect of pentaalginate oligosaccharide derivatives on Electrical stimulation of trigeminal ganglion migraine-causing rats

1 materials of the experiment

1.1 drugs and reagents

The preparation method of the second example is adopted to prepare the algin oligosaccharide (n is 2-8, namely the components 4-10 in the figure) mixture of the formula (II). The mixture is white powder, and is easily soluble in water; (ii) a Evans blue, paraformaldehyde, sucrose, glycerol, hydrogen peroxide, Triton X-100, DAB were all purchased from SIGMA; rabbit anti-murine c-fos antibody (Ab-5), Oncogene research products, USA; biotin-labeled goat anti-rabbit IgG serum, Alexis, Switzerland; avidin biotinperoxidase complex, Vector Laboratories Inc., Burlingame, Calif., USA.

1.2 Experimental animals

SD rats, 5 months old, male, with a body weight of 200-240 g, were provided by Shanghai Sphere-BikKa laboratory animals Co.

1.3 Experimental instruments

An LZ-6 drill rig, a new medical instrument factory in Shanghai old port; brain stereotaxic apparatus, STOELT ING; MD2000 stimulator, university of medical, tokyo.

2 test method

2.1 animal grouping and administration

Male SD rats were randomly divided into a blank control group, a sham operation group, a model group, and alginate oligosaccharide derivatives 12.5, 25, 50, 100, 200mg/kg groups, each of which contained 10 animals. The corresponding drugs are orally taken to each group, and distilled water is orally taken to a blank control group, a sham operation group and a model group. After continuous administration for 10 days, except for a blank control group, after all rats are anesthetized by intraperitoneal injection of 350mg/kg chloral hydrate, the rats are fixed on a stereotaxic instrument, a central incision is made on the vertex of the head, skin and muscle are cut layer by layer, and skull cap bones are exposed at a middle opening of a sagittal suture of the brain. After 3 mm of posterior movement of the anterior chimney and 3 mm of lateral movement, the electrode was inserted into the trigeminal ganglion (9.5 mm deep from the dura) using a dental drill, and then the operation was continued with anesthesia. All manipulations were performed under sterile conditions. And (3) debugging a stimulation electrode, wherein the electrical stimulation parameters are 200ms in period, 10v in amplitude, 5ms in wave width and 10 minutes in stimulation. Sham animals only inserted electrodes and were not stimulated. The injection of 50mg/kg Evans blue is injected into the right femoral vein 7 minutes before stimulation, and the perfusion is fixed within 20 minutes after the stimulation is finished.

5 minutes after stimulation is finished, perfusing the left ventricle for 2 minutes, opening the skull, taking the whole brain, fixing, and measuring c-fos by immunohistochemistry of pathological sections; and (3) additionally determining the position of the electrode, separating the insertion position of the electrode and the dura mater at the corresponding position of the other cerebral hemisphere, washing by deionized water, then spreading on a glass slide, drying at 37 ℃ for 15 minutes, and fixing by 70% glycerol. Fluorescence intensities of the specified areas of the stimulation side and the control side are detected on a confocal microscope at an excitation wavelength of 647nm and an emission wavelength of 680nm, and the ratio of the fluorescence intensities of the stimulation side and the control side is calculated to indicate the Plasma Protein exudation rate (PPE). The whole brain is continuously frozen into coronal sections with the thickness of 10 μm, and immunohistochemical fluorescence marks c-fos positive cells. Under a confocal microscope, randomly selecting 5 visual fields, determining the number of positive cells on the experimental side and the control side of the tail side of the trigeminal spinal nucleus, and then taking the average value of the 5 visual fields to obtain the average number of the positive cells.

3 statistical treatment

The results were statistically analyzed and expressed as "mean ± standard deviation" (mean ± SD) and compared using analysis of variance (AN0 VA). The difference is significant when p is less than 0.05, and the difference is very significant when p is less than 0.01.

4 results of the test

Activation of the trigeminal vasculature is a key component in the development of pain in migraine sufferers, where the development of neurogenic inflammation of the meninges plays an important role in the development and maintenance of pain in migraine. When trigeminal nerve distributed in dura mater is stimulated, vasoactive substances are released, and meningeal vasodilatation, plasma component extravasation, mast cell degranulation and platelet activation are caused to produce migraine. In addition, the neurotransmitter released after the pain stimulation is combined with the corresponding receptor on the cell membrane, under the action of the second messenger, the C-fos mRNA gene is expressed, and is translated and synthesized into the C-fos protein in the cell nucleus, so that the physiological effect on the body for a long time is generated. Therefore, in migraine, the number of cells expressing C-fos mRNA and C-fos protein in the trigeminal spinal nucleus and nucleus raphanus are increased. Therefore, the degree of migraine can be reflected by measuring the amount of serum protein exudation in the dura mater of the migraine animal and the number of c-fos positive cells on the caudal side of the spinal nucleus of the trigeminal nerve.

The research results are shown in Table 5, the electrical stimulation of the trigeminal ganglion of the rat obviously causes the exudation of the serum protein of the dura mater, the alginate oligosaccharide derivatives 25, 50, 100 and 200mg/kg are continuously administrated for 10 days, the exudation of the serum protein of the dura mater caused by the electrical stimulation of the trigeminal ganglion of the rat can be obviously inhibited, and the inhibition rates are respectively 43.1%, 59.7%, 79.2% and 97.2%; the number of the positive cells of the c-fos expression at the tail side of the spinal nucleus of the trigeminal nerve of the electrically stimulated rat is obviously increased, and the increase of the positive cells of the c-fos expression can be obviously inhibited by continuously administering the alginate oligosaccharide derivatives for 10 days at 25, 50, 100 and 200mg/kg, wherein the inhibition rates are respectively 61.1%, 64.6%, 81.7% and 89.3%. The results show that the alginate oligosaccharide derivative can obviously inhibit rat migraine caused by electrically stimulating trigeminal ganglia. This indicates that the alginate oligosaccharide derivatives have an effect of treating migraine and also indicates that the alginate oligosaccharide derivatives have an effect of treating pain.

TABLE 5 Effect of alginate oligosaccharide derivatives on the electrical stimulation of the Leptocephalia trigeminal ganglia rat dura mater serum protein exudation rate (PPE rate) and the number of c-fos positive cells at the caudal side of the trigeminal spinal nucleus (n 10, mean + -SD)

Blank control group with # p less than 0.01 vs; p < 0.05, p < 0.01 vs model set

Claims

1. The application of the algin oligosaccharide and the derivatives thereof or the pharmaceutically acceptable salts of the algin oligosaccharide and the derivatives thereof in preparing the drugs for treating pains, wherein the algin oligosaccharide and the derivatives thereof are the algin oligosaccharide represented by the following structural formula (II) and the derivatives thereof or the pharmaceutically acceptable salts of the oligosaccharide and the derivatives thereof,

wherein n in formula (II) is one or more integers selected from 0 to 8,

wherein the pain is acute pain, chronic pain, neuropathic pain, post-operative pain, chronic lower back pain, cluster headache, herpetic neuralgia, phantom limb pain, central pain, dental pain, opioid-resistant pain, visceral pain, surgical pain, bone injury pain, pain during exertion and childbirth, pain due to burns including sunburn, postpartum pain, migraine, angina and pain associated with the genitourinary tract (including cystitis), vascular pain, trigeminal neuralgia, intercostal fascia pain, surgical incision pain, chronic inflammatory pain, heel pain, muscle pain, bone pain, joint pain, cancer pain, non-cancer pain.

2. The use according to claim 1, wherein n in formula (II) is one or more integers selected from 2 to 8.

3. The use according to claim 1 or 2, wherein n in formula (II) is 4.