Analgesic composition

Abstract

To provide an analgesic composition that can substitute for NSAIDs.SOLUTION: An analgesic composition contains microalgae as an active ingredient.SELECTED DRAWING: Figure 3B

Description

[Technical Field]

[0001] The present invention relates to an analgesic composition. [Background technology]

[0002] For example, Japanese Patent Application Laid-Open Publication No. 2021-013313 (Patent Document 1) discloses microalgae of the Pavlovaceae family that contain high concentrations of fucoxanthin, dietary fiber, eicosapentaenoic acid, gamma (γ)-aminobutyric acid, etc. Recently, the nutritional components contained in microalgae (such as chlorella and euglena) have attracted attention, and efforts are underway to use them in health foods, food ingredients, etc. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent Publication No. 2021-013313 Summary of the Invention [Problem to be solved by the invention]

[0004] Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used drugs expected to have anti-inflammatory and analgesic effects, but there are concerns about side effects on the digestive tract, such as gastritis, and the possibility of various adverse effects on the renal and cardiovascular systems. For this reason, there is a strong demand for new analgesic materials that can replace NSAIDs or reduce the dosage of NSAIDs to suppress side effects, etc. However, for example, Patent Document 1 mentioned above does not mention whether microalgae of the Pavlovaceae family have anti-inflammatory and analgesic effects.

[0005] In view of the above circumstances, an object of the present invention is to provide an analgesic composition that can replace NSAIDs. [Means for solving the problem]

[0006] The present inventors conducted extensive research to solve the above problems and arrived at the present invention. The inventors focused on microalgae of the Pavlovaceae family, particularly microalgae of the genus Pavlova, and investigated whether they possess analgesic effects that can replace NSAIDs. As a result, they discovered that the microalgae at least have the effect of suppressing thermal hyperalgesia, leading to the completion of the present invention. The present invention relates to an analgesic composition as described below.

[0007] [1] A pain-relieving composition containing microalgae as an active ingredient. [2] The analgesic composition according to [1], wherein the microalgae are classified into the genus Pavlova. [3] The analgesic composition according to [1] or [2], which is an antihyperalgesic agent or an antiallodynic agent. [4] The analgesic composition according to any one of [1] to [3], which is a therapeutic agent for arthralgia. [5] The analgesic composition according to any one of [1] to [4], which is a c-FOS activity inhibitor. [6] The analgesic composition according to any one of [1] to [5], further comprising a nonsteroidal anti-inflammatory agent. [7] The analgesic composition according to any one of [1] to [6], wherein the microalgae is administered orally to a living body at a daily dose of 5 mg / kg to 300 mg / kg. [8] The analgesic composition according to [7], which is used together with a nonsteroidal anti-inflammatory agent. [9] A food or beverage material comprising the analgesic composition according to any one of [1] to [8].

[10] The food and beverage material according to [9], which is applied to at least one selected from the group consisting of food and beverage products, oral medicines, and animal feed. [Effects of the Invention]

[0008] According to the present invention, an analgesic composition that can replace NSAIDs is provided. [Brief explanation of the drawings]

[0009] [Figure 1A] FIG. 1A is a line graph showing the relationship between the amount of DFC administered to arthritis model mice and its antihyperalgesic effect. [Figure 1B] FIG. 1B is a bar graph showing the relationship between the area under the curve for the antihyperalgesic effect from immediately after DFC administration to 24 hours later, calculated from the graph in FIG. 1A, and the dose. [Figure 2A] FIG. 2A is a line graph showing the comparative antihyperalgesic effects of administering low-dose DFC alone to arthritis model mice and administering low-dose DFC in combination with Pavlova. [Figure 2B] Figure 2B is a bar graph comparing the areas under the curve for the antihyperalgesic effect of low-dose DFC alone or in combination with Pavlova from immediately after administration to 24 hours after administration, calculated from the graph in Figure 2A. [Figure 3A] Figure 3A shows fluorescent images of the analgesic mechanism of Pavlova by examining the changes in the number of c-FOS-positive cells in the spinal cord of arthritis model mice. [Figure 3B] FIG. 3B is a bar graph showing the evaluation results based on the image of FIG. 3A. DETAILED DESCRIPTION OF THE INVENTION

[0010] Hereinafter, embodiments of the present invention (hereinafter also referred to as "present embodiments") will be described, but the present invention is not limited thereto. Herein, in this specification, an expression in the form of "A to B" means the upper and lower limits of a range (i.e., A or more and B or less), and when no unit is specified for A and a unit is specified only for B, the unit of A and the unit of B are the same.

[0011] [Analgesic composition] The analgesic composition according to this embodiment contains microalgae as an active ingredient. As described below, an analgesic composition with these characteristics can replace NSAIDs by utilizing the analgesic effects of the microalgae. Alternatively, when used in combination with NSAIDs, the analgesic composition may potentially reduce the dosage of the NSAIDs and suppress side effects. As used herein, "microalgae" refers to microorganisms containing chloroplasts and measuring, for example, 0.1 μm to 1 mm in microscopic size, and generally refers to algae of this size that live in water. Microalgae include organisms belonging to the prokaryotic phylum Cyanobacteria and the eukaryotic phylum Glaucophyta, Rhodophyta (red algae), Chlorophyta, Cryptophyta (cryptophytes), Haptophyta (haptophytes), Heterokontophyta, Dinophyta (dinoflagellates), Euglena, and Chlorarachniophyta.

[0012] Of these, the phylum Haptophyta (haptophytes) includes the class Haptophyceae. Haptophytes are phytoplankton with cell diameters of approximately 5 to 50 μm, and are autotrophic organisms that perform photosynthesis. The class Haptophyceae includes the subclass Pavloviales and the subclass Prymnesiophyceae. The subclass Pavloviales includes the order Pavlovales. The order Pavlovales includes the family Pavlovaceae. The family Pavlovaceae includes the genus Pavlova. The genus Pavlova includes P. calceolate, P. granifera, P. gyrans, P. lutheri, P. pinguis, and P. salina.

[0013] The microalgae are preferably classified into the genus Pavlova. In this preferred embodiment, the microalgae are more preferably P. granifera or P. gyrans. The P. granifera and P. gyrans have been deposited, and the NBRC accession number for P. granifera is NBRC 114066. The NBRC accession number for P. gyrans is NBRC 102809. Specifically, the microalgae may be the Pavlova OPM S30543 strain (the algae strain identified by the accession number NBRC 114066, the P. granifera), or the Pavlova OPM S30543X strain (the algae strain identified by the accession number NBRC 102809, the P. gyrans), or a derivative thereof. As used herein, the term "derivative" refers to a strain containing a gene that contains a region substantially homologous to the DNA of the microalgae of interest. When such a strain is aligned using a computer homology program known in the art and compared with the whole genome sequence of the original strain, it has a whole genome sequence that is at least 30% identical, preferably 60% identical, more preferably 90% identical, and most preferably 95% identical or 99% identical. However, as used herein, the term "a strain derived from Pavlova OPM S30543 or the like" does not necessarily mean that the strain was derived from Pavlova OPM S30543 or the like, but rather means a microalga that contains a gene containing a region substantially homologous to the DNA of the target microalgae and that exhibits at least some of its biological activity.

[0014] As used herein, the term "biological activity" refers to various functions (e.g., analgesic activity (specifically, antihyperalgesic activity, antiallodynic activity, etc.)) that can be exhibited by organisms, including microalgae, in a certain environment. Such biological activity can be measured by techniques well known in the art. For example, the biological activity can be measured qualitatively and quantitatively by evaluating the response of the organism to a given exposure or stimulus. For example, the biological activity of the microalgae can be evaluated by an increase or decrease in the amount of upstream or downstream proteins, or an improvement or decrease in other functions, after some stimulus or event.

[0015] The present inventors focused on the microalgae of the genus Pavlova described above and investigated whether they possess analgesic effects that can replace NSAIDs. As a result, as will be described in the Examples below, it was discovered that the microalgae exhibit at least an unknown attribute of suppressing thermal hyperalgesia, making them suitable for use in the new application of analgesia. Here, the microalgae contained as an active ingredient in the analgesic composition may typically be the algae bodies of the microalgae, or a dried product containing some of the components contained in the algae bodies, or a product further processed from the dried product.

[0016] When the microalgae contained as an active ingredient in the analgesic composition is a dried product, the moisture content of the microalgae may be 50% by mass or less, 10% by mass or less, 5% by mass or less, 3% by mass or less, 1% by mass or less, 0.5% by mass or less, 0.2% by mass or less, 0.1% by mass or less, or 0.05% by mass or less. The analgesic composition may be provided by immersing the dried microalgae or a purified product thereof in oil or encapsulating it in a capsule (e.g., a soft capsule). In this manner, the analgesic composition can be stabilized.

[0017] The analgesic composition may contain the microalgae in any amount, but may contain 0.01 to 100% by mass. The analgesic composition may contain the microalgae in an amount of 0.01% or more, 0.1% or more, 1% or more, 5% or more, 10% or more, 20%, 50%, 80% or more, or even 100% by mass. The analgesic composition may contain the microalgae in an amount of 100% or less, 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, or 1% or less by mass. The analgesic composition can contain any combination of flavoring agents, odorants, masking agents, etc. to adjust the taste or flavor of the microalgae when ingested. The analgesic composition can also adjust the taste or flavor of the microalgae when ingested by means of coating or encapsulation, etc.

[0018] <Antihyperalgesic / Antiallodynic> The analgesic composition according to this embodiment is preferably an antihyperalgesic agent or an antiallodynic agent. The analgesic composition is preferably an analgesic, an anti-inflammatory agent, or a neuropathic pain agent. As used herein, "hyperalgesia" refers to a state in which pain is enhanced in response to stimuli exceeding the pain threshold. The term "hyperalgesia" is used to refer to the hyperreactivity observed in cases involving a normal pain threshold and an increase therein, such as in neuropathic pain. Therefore, the suppressive effect of the antihyperalgesic agent on hyperalgesia suppresses the enhancement of pain in response to stimuli exceeding the pain threshold.

[0019] In this specification, "allodynia" refers to a state in which pain is induced by tactile stimuli, mild pressure stimuli, moderate warm or cold stimuli, etc. on clearly normal skin. "Allodynia" is also called allodynia. "Allodynia" is distinguished from the above-mentioned "hyperalgesia" in that it can induce pain in response to stimuli that do not exceed the pain threshold. Therefore, the action of the anti-allodynic agent can suppress the induction of pain in response to stimuli that do not exceed the pain threshold.

[0020] Since the above analgesic composition is an anti-hyperalgesic agent or an anti-allodynic agent, it may be applicable to cases and symptoms where the analgesic effect of NSAIDs is not sufficient or hardly expected, such as neuropathic pain.

[0021] <Therapeutic agent for joint pain> The analgesic composition according to this embodiment is preferably a therapeutic agent for joint pain. Typically, the above analgesic composition is preferably an arthritis therapeutic agent. The above arthritis therapeutic agent may be applied, for example, to osteoarthritis. The above analgesic composition can obtain an effect of reducing joint pain, for example, through the action of suppressing the above-mentioned hyperalgesia or through the action of suppressing the induction of pain in response to stimuli that do not exceed the pain threshold.

[0022] Since the above analgesic composition is a therapeutic agent for joint pain, it may be preferably applicable as an alternative to NSAIDs for chronic pain such as osteoarthritis. This is because long-term administration of NSAIDs tends to be avoided due to concerns about side effects and the like for chronic pain. Furthermore, the above analgesic composition may be preferably applicable as an alternative to NSAIDs for chronic low back pain, which is known as a chronic pain disease other than osteoarthritis.

[0023] <c-FOS activity inhibitor> The analgesic composition according to this embodiment is preferably a c-FOS activity inhibitor. c-FOS is a phosphorylated protein localized in the cell nucleus and is used as a marker indicating the activation of nerve activity. When nerve activity is enhanced in a living body, the expression of c-FOS is induced in nerve cells, and the number of c-FOS-positive nerve cells increases. Along with this, the number of activated nerve cells increases, and there is a possibility that hyperalgesia or allodynia may occur as neuropathic pain. Therefore, it is possible that hyperalgesia or allodynia can be suppressed by the action of a c-FOS activity inhibitor.

[0024] Since the analgesic composition is a c-FOS activity inhibitor, it can suppress the above-mentioned hyperalgesia or allodynia, and can be applied to cases or symptoms where, for example, the analgesic effect of NSAIDs for neuropathic pain and the like is not sufficient or almost no effect can be expected.

[0025] <Combined use with NSAIDs> The analgesic composition according to this embodiment may contain a non-steroidal anti-inflammatory drug (NSAIDs). In this aspect, the amount of NSAIDs used expecting an analgesic effect can be reduced compared to the amount when used alone. For example, the amount of NSAIDs contained in the above analgesic composition can be 50% or less, 25% or less, or 10% or less per day on a mass basis compared to the amount when used alone, preferably 5% or less per day, more preferably 3% or less per day, and even more preferably 2% or less or 1% or less per day. Thereby, it is possible to reduce the dosage of NSAIDs and suppress its side effects, etc., while maintaining the analgesic effect or efficacy by the above analgesic composition.

[0026] The NSAIDs contained in the above analgesic composition are not particularly limited, and examples thereof include aspirin, acetaminophen, ibuprofen, diclofenac, or their derivatives.

[0027] <Use, dosage form, and dosage> The analgesic composition according to this embodiment is preferably administered orally. This allows the analgesic composition to exhibit a favorable analgesic effect. The daily dose of the microalgae administered to a living body is not particularly limited, but is preferably 1 mg / kg or more, 10 mg / kg or more, 20 mg / kg or more, 30 mg / kg or more, 40 mg / kg or more, 50 mg / kg or more, 60 mg / kg or more, 70 mg / kg or more, 80 mg / kg or more, 90 mg / kg or more, or 100 mg / kg or more. It may also be 10 g / kg or less, 5 g / kg or less, 1000 mg / kg or less, 900 mg / kg or less, 800 mg / kg or less, 700 mg / kg or less, 600 mg / kg or less, 500 mg / kg or less, 400 mg / kg or less, or 300 mg / kg or less. Therefore, the daily dose of the microalgae administered to a living body is 1 mg to 10 g / kg, and the composition may be administered orally. The daily dose of the microalgae to a living organism may be 10 to 1000 mg / kg, 20 to 900 mg / kg, 30 to 800 mg / kg, 40 to 700 mg / kg, 50 to 500 mg / kg, 60 to 400 mg / kg, 70 to 350 mg / kg, 100 to 300 mg / kg, 100 to 1000 mg / kg, 150 to 700 mg / kg, 200 to 500 mg / kg, etc. The daily dose of the microalgae to a living organism may be 5 mg / kg body weight or more and 300 mg / kg body weight or less.

[0028] The desired dosage of the analgesic composition can be increased or decreased as appropriate, taking into consideration the method of administration, the age and sex of the patient, the severity of symptoms, and the like. For example, the analgesic composition is preferably administered in an amount of 1 mg or more, 10 mg or more, 20 mg or more, 30 mg or more, 40 mg or more, 50 mg or more, 60 mg or more, 70 mg or more, 80 mg or more, 90 mg or more, 100 mg, 300 mg, 500 mg, 700 mg, 1000 mg, 1200 mg, 1300 mg, 1500 mg, 2000 mg, or 2500 mg or more per day for an adult, but may also be administered in an amount of 1000 g or less, 200 g or less, 100 g or less, 10 g or less, 5000 mg or less, 4000 mg or less, 3000 mg or less, 2000 mg or less, 1000 mg or less, 900 mg or less, 800 mg or less, 700 mg or less, 600 mg or less, 500 mg or less, 400 mg or less, or 300 mg or less. The daily dose of the microalgae for an adult may be 1 to 10,000 mg, 10 to 5,000 mg, 20 to 3,000 mg, 500 to 3,000 mg, 1,000 to 2,000 mg, 10 to 1,000 mg, 40 to 700 mg, 50 to 500 mg, 60 to 400 mg, 70 to 350 mg, 100 to 300 mg, etc.

[0029] The above-mentioned "living body" includes not only humans as mammals but also animals, particularly pets, etc. The dose of the analgesic composition can be determined appropriately for ingestion by these living bodies. When the analgesic composition is used as feed, pet food, or the like, the target organisms are not particularly limited, but are preferably mammals, reptiles, amphibians, birds, or fish, and more preferably mammals other than humans, such as platypus, echidna, opossum, quoll, kangaroo, aardvark, rock badger, elephant, armadillo, sloth, anteater, tree shrew, flying lemur, chimpanzee, rabbit, degu, dormouse, squirrel, raccoon, mouse, hedgehog, chinchilla, ferret, camel, wild boar, giraffe, deer, cow, goat, hippopotamus, whale, dolphin, horse, rhinoceros, tapir, bat, monkey, tiger, wolf, weasel, bear, seal, dog, cat, parakeet, parrot, finch, owl, and horned owl, and more preferably dog or cat.

[0030] When the analgesic composition is used as feed, pet food, or the like, it may be given in divided doses several times a day by adding it to staple food, or it may be given as a snack at any time.

[0031] Here, the concept of the dosage (dose) of the analgesic composition for an animal (living body) will be explained using a mouse as an example as follows: That is, when converting the dosage (dose) for a mouse to the dosage (dose) for a living body, the concept of the human equivalent dose can be used, although it is not limited thereto. Bioequivalent dose (HED) = mouse dose (mg / kg) × (mouse body weight (kg) / animal body weight (kg)) 0.33

[0032] The biological equivalent dose is calculated using the formula above. For example, for a mouse weighing 30 g and dosed at 300 mg / kg, the HED is calculated as 24.4 mg / kg. For a 60 kg human, the HED is calculated as approximately 1500 mg / human, and for a 10 kg dog, it is calculated as approximately 440 mg / dog.

[0033] The following concept can be used to convert the dose for mice to that for other living organisms: For example, if a mouse takes about 300 mg / kg of body weight of the above analgesic composition per day, the intake per kg of body weight is calculated as follows: 2 To convert this to an intake per person, the factor listed in Table 1 of the FDA guidance (Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers, July 2005) is used. According to this, the factor for mice is multiplied by 3, resulting in a daily intake of 900 mg / m2 for mice. 2 This means that the patient has ingested an analgesic composition equivalent to 1000 mg of body surface area. 2When converting the intake amount per kg of body weight for each animal species, for humans it is divided by the factor of 37 specified in the FDA guidance above, which gives 24.3 mg / kg body weight per day, for dogs it is divided by the factor of 20 specified in the FDA guidance above, which gives 45.0 mg / kg body weight per day, and for rabbits it is divided by the factor of 12 specified in the FDA guidance above, which gives 75.0 mg / kg body weight per day.

[0034] The analgesic composition according to this embodiment is preferably used in combination with a nonsteroidal anti-inflammatory drug (NSAID). In such applications, the amount of NSAID used to achieve an analgesic effect can be reduced compared to the amount when the NSAID is used alone. For example, the amount of NSAID used in combination with the analgesic composition can be 50% or less, 25% or less, or 10% or less per day, by mass, compared to the amount when the NSAID is used alone, preferably 5% or less per day, more preferably 3% or less per day, and even more preferably 2% or less, or 1% or less per day. This may allow the analgesic composition to reduce the dose of NSAIDs and suppress their side effects while maintaining analgesic activity or effectiveness.

[0035] The analgesic composition is expected to be primarily administered orally as described above, but it is not excluded that it may also be in a dosage form for parenteral administration, such as an injection, suppository, or topical skin preparation. For example, topical skin preparations include patches, tapes, creams, lotions, lotions, emulsions, foundations, packs, foams, plasters, ointments, poultices, and aerosols, and analgesic effects can also be expected in these cases.

[0036] [Materials for food and beverages] The food and beverage material according to this embodiment includes the analgesic composition. A food and beverage material having such characteristics can replace NSAIDs by utilizing the analgesic effect of the microalgae in the analgesic composition. Alternatively, when used in combination with NSAIDs, the food and beverage material may be able to reduce the dosage of NSAIDs and suppress side effects. The food and beverage material is preferably applied to at least one selected from the group consisting of food and beverages, oral medications, and animal feed. The food and beverage material can also be mixed with regular meals or feed. The food and beverage material can also be used as a specified health food or a functional food.

[0037] As used herein, "food and beverage products" refers to products intended for ingestion by animals (including humans). Food and beverage products include not only commonly used foods and beverages, but also food additives, functional foods (e.g., foods for specified health uses, foods with functional claims, foods with nutrient functions, etc.), and supplements. Furthermore, as used herein, "animal feed" refers to products intended for ingestion by pet animals or livestock, among others.

[0038] As used herein, "drugs" in "oral medications" refers to drugs administered to humans or animals for the diagnosis, treatment, or prevention of disease. This term includes items listed in the Japanese Pharmacopoeia, items intended for use in the diagnosis, treatment, or prevention of disease in humans or animals that are not mechanical instruments, dental materials, medical supplies, or sanitary products, and items intended to affect the structure or function of the human or animal body that are not mechanical instruments, dental materials, medical supplies, or sanitary products. Furthermore, as used herein, "oral medications" also includes quasi-drugs. "Quasi-drugs" are defined in Japan's "Act on Ensuring Quality, Efficacy, and Safety of Pharmaceuticals, Medical Devices, Gene Therapy Products, and Gene Therapy Products," and are classified as intermediates between pharmaceuticals and cosmetics. These include products with mild effects on the human body, including mechanical instruments with mild effects on the human body. Examples of quasi-drugs include, but are not limited to, designated quasi-drugs (such as supplements and some gastrointestinal medications). As used herein, "oral" in "oral medications" refers to the above-mentioned medications that are ingested orally by animals (including humans).

[0039] When the food or beverage material is an "oral pharmaceutical," the daily oral intake or dosage may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form.

[0040] When the food or beverage material is an "oral pharmaceutical," the oral pharmaceutical can be taken or administered once to several times a day, typically 1 to 6 times a day, 1 to 3 times a day, 1 to 2 times a day, or for any period, interval, or as needed, but 3 times a day or as needed is preferred.

[0041] Use of the analgesic composition The analgesic composition according to the present embodiment can be added to or mixed with foods, beverages, pharmaceuticals, feed, or pet food. The analgesic composition can also be used as is in foods, beverages, pharmaceuticals, feed, or pet food. The analgesic composition can be used as a food or beverage that explicitly or implicitly claims analgesia, anti-inflammatory properties, or anti-hyperalgesia as its functional properties, i.e., health foods, functional foods, foods for patients, and foods for specified health uses. The analgesic composition can also be used as so-called doctor's supplements recommended or offered by doctors in internal medicine or orthopedic departments in hospitals and / or clinics, veterinary clinics, etc., even if the functional properties are not explicitly or implicitly claimed. When the functional properties are explicitly claimed, they may be stated as, but are not limited to, alleviating temporary pain in daily life or alleviating transient pain caused by daily life or exercise. It is also possible to claim that prior intake of the analgesic composition can prevent pain from occurring or reduce the severity of pain.

[0042] Health foods, functional foods, foods for patients, and foods for specified health uses can be used in various dosage forms, such as solid preparations (tablets, orally disintegrating tablets, granules, fine granules, powders, capsules, chewable tablets, candy, etc.), liquid preparations (syrups, suspensions), and liquid foods. Food preparations can be produced in the same manner as known pharmaceutical preparations, by mixing the active ingredient with a food-acceptable carrier, such as a suitable excipient, and then producing the preparation using conventional means. The dosage form is not limited, but is preferably an orally disintegrating tablet, chewable tablet, candy, granules, powder, or liquid, from the viewpoint of significantly exhibiting the effects of the analgesic composition.

[0043] For example, tablets can be prepared by mixing a powdered active ingredient with a pharmaceutically acceptable carrier (such as an excipient) and compressing the mixture, and confectionery tablets such as candies can be prepared by pouring the mixture into a mold. Tablets can also be sugar-coated to form sugar-coated tablets. Furthermore, tablets can be single-layer tablets or layered tablets such as double-layer tablets.

[0044] Powdered granules such as granules may be prepared by various granulation methods (extrusion granulation, milling granulation, dry compaction granulation, fluidized bed granulation, tumbling granulation, high-speed stirring granulation, etc.), and tablets can be prepared by an appropriate combination of the above-mentioned granulation methods and tableting methods (wet tableting, direct tableting), etc.

[0045] Capsules can be prepared by filling a capsule (soft or hard capsule) with powder (powder, granules, etc.) by a conventional method.

[0046] A liquid preparation can be prepared by dissolving or dispersing each component in an aqueous medium (purified water, purified water containing ethanol, etc.) which is a carrier component, filtering or sterilizing the resulting solution as necessary, filling the resulting solution into a predetermined container, and sterilizing the resulting solution. The preferred dosage form of the solid preparation is a capsule or tablet, and soft capsules (soft capsules) are more preferred.

[0047] Soft capsules are preferred by users because they have a smooth surface and are easy to swallow. Common methods for manufacturing soft capsules include the flat plate method, the rotary method, and the seamless method.

[0048] In the rotary method (punching method), a sheet-like capsule shell sandwiches the flowing filling material and forms a capsule shape along the holes in a rotating cylindrical mold. On the other hand, in the seamless method (dropping method), the capsule shell composition and the filling material are simultaneously ejected from multiple concentric nozzles to form a seamless capsule shape.

[0049] The base material for the soft capsule shell is not particularly limited, but may be starch, pullulan, cellulose, polyvinyl alcohol, gelatin, succinated gelatin, etc., with starch, gelatin, and succinated gelatin being preferred, and gelatin and succinated gelatin being more preferred. These may be used alone or in combination of two or more.

[0050] The analgesic composition can also be used in liquid beverages such as soups, juices, fruit juice drinks, milk, dairy drinks, whey drinks, lactic acid bacteria drinks, tea drinks, alcoholic drinks, coffee drinks, carbonated drinks, soft drinks, water drinks, cocoa drinks, jelly drinks, sports drinks, and diet drinks; semi-solid foods such as pudding and yogurt; noodles; confectioneries; spreads; and the like.

[0051] When the analgesic composition is prepared as a food composition, various food additives may be added, such as antioxidants, colorings, flavorings, seasonings, sweeteners, acidulants, pH adjusters, quality stabilizers, and preservatives.

[0052] When the analgesic composition is prepared as a pharmaceutical composition, it is prepared as a formulation containing the microalgae as an active ingredient and preferably a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier generally refers to an inert, non-toxic, solid or liquid filler, diluent, or encapsulating material that does not react with the active ingredient, such as water, ethanol, polyols, suitable mixtures thereof, vegetable oils, and other solvents or dispersion media.

[0053] The pharmaceutical composition is administered orally or parenterally, for example, into the oral cavity, the digestive tract, or the nasal cavity. Orally administered formulations include solid formulations (tablets, orally disintegrating tablets, granules, fine granules, powders, capsules, chewable tablets, lozenges, etc.) and liquid formulations (syrups, suspensions, inhalants). Parenterally administered formulations include eye drops, drip infusions, nasal drops, and injections. The formulation form is not limited, but is preferably an orally disintegrating tablet, chewable tablet, lozenge, granule, powder, or liquid formulation, from the viewpoint of significantly exhibiting the effects of the analgesic composition.

[0054] The pharmaceutical composition may further contain additives commonly used in the pharmaceutical field. Such additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, coloring agents, flavoring agents, etc., and can be used appropriately as needed. To achieve sustained release so as to ensure long-term action, the composition may also be coated with a known retardant, etc. The pharmaceutical composition may further contain other additives or drugs, such as antacids and gastric mucosa protectants, as needed.

[0055] The pharmaceutical composition can be applied in the form of an oral composition, an internal composition, etc. The pharmaceutical composition may be used therapeutically or non-therapeutically. The analgesic composition can also be mixed in the above-mentioned dosage form with the food and beverage material described below. [Example]

[0056] The present invention will be described in more detail below with reference to several examples, but the present invention is not limited thereto. All examples described below were conducted in accordance with the animal experiment regulations of Tohoku Medical and Pharmaceutical University.

[0057] [First Example] <Sample preparation> The sample used in this example (analgesic composition, hereinafter also referred to as "Pavlova") was prepared by obtaining a dried product of microalgae (trade name: "Pavlova", manufactured by Rohto Pharmaceutical Co., Ltd.). This sample consisted of 100% by mass of the microalgae, which belonged to the genus Pavlova. In this example, the sample was suspended in a 0.5% by mass CMC (sodium carboxymethylcellulose, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) solution.

[0058] <Preparation of animals> The animals used in this and other examples were prepared by obtaining young adult ddy mice (7-12 weeks old, weighing 25-40 g, manufactured by Japan SLC Co., Ltd.). Approximately equal numbers of male and female mice were used in each example. Until their use in each example, the mice were housed in a controlled environment at 22±2°C, 55±10% relative humidity, and a 12-hour light / dark cycle (light period 7:00-19:00). Four mice were housed in each plastic cage (30 cm length × 20 cm width × 15 cm height). The mice were fed solid feed (product name: "FR-2", manufactured by Funabashi Farm Co., Ltd.) and tap water ad libitum.

[0059] <Verification of the effect of Pavlova on thermal hyperalgesia> To evaluate the analgesic effect of Pavlova on thermal hyperalgesia in arthritis model mice, Pavlova or CMC (Fujifilm Wako Pure Chemical Industries, Ltd.) was orally administered to the mice for 28 days at a dose of 300 mg / kg / day.

[0060] (Creation of arthritis model mice) On day 14 after the oral administration of Pavlova or CMC, 50% by mass complete Freund's adjuvant (CFA, Invitrogen) was administered periarticularly in the amounts described below to generate arthritis model mice. Specifically, the arthritis model mice were generated using a modified version of the method described by Montilla-Garcia et al. Specifically, under inhalation anesthesia with isoflurane (Fujifilm Wako Pure Chemical Corporation) / oxygen, 50% by mass CFA diluted with incomplete Freund's adjuvant (IFA, Invitrogen) was administered periarticularly around the right tibial joint to induce arthritis. 15 μL of 50% by mass CFA was administered intraarticularly and periarticularly (30 μL total). The control group of mice received the same volume (i.e., 30 μL) of saline instead of 50% by mass CFA.

[0061] (Method for measuring thermal hyperalgesia) Thermal hyperalgesia was measured using the Hargreaves method. Specifically, using a Hargreaves Analgesia Meter 390 (IITC Life Sciences), the mice were placed individually in each compartment (9 cm × 5 cm × 5 cm high) on the glass plate of the instrument and allowed to acclimate for at least 1 hour. After confirming that the mice had ceased to move spontaneously, a halogen lamp was projected onto the plantar surface of the right hind paw from beneath the glass plate, and the time until the mice exhibited a withdrawal response (latency) was measured. A higher latency indicated a longer time until the mice exhibited a withdrawal response, indicating that thermal hyperalgesia was not enhanced (pain was suppressed).

[0062] The output of the halogen lamp light source was set to 20% of the maximum output of the device. Because mouse urine staining of the glass plate would affect the measurement values, the urine was immediately wiped off to keep the measurement environment clean. To avoid tissue damage to the mouse, the cutoff time was set to 40 seconds. Response latencies were calculated as the average of three measurements.

[0063] (evaluation) The reaction latencies of the Pavlova- or CMC-administered mice (arthritis model mice) were measured using the method described above on the first day of Pavlova or CMC administration (baseline), the day before 50% CFA or saline administration (Day -1), the day after 50% CFA or saline administration (Day +1), 3 days after 50% CFA or saline administration (Day +3), 7 days after 50% CFA or saline administration (Day +7), and 14 days after 50% CFA or saline administration (Day +14). Eight mice were used in each group. The results are shown in Table 1.

[0064] In Table 1, "0.5% CMC / Control" represents mice administered with CMC during the test period (28 days) and used as the control group (mice administered with saline). "0.5% CMC / 0.5% CFA" represents arthritis model mice administered with CMC during the test period (28 days) (mice administered with CFA). "Pav 300 mg / kg / Control" represents mice administered with Pavlova during the test period (28 days) and used as the control group (mice administered with saline). "Pav 300 mg / kg / 0.5% CFA" represents arthritis model mice administered with Pavlova during the test period (28 days) (mice administered with CFA).

[0065] [Table 1]

[0066] <Consideration> According to Table 1, the "Pav 300 mg / kg / 0.5% CFA" group had longer response latencies on Day +1, Day +3, Day +7, etc. compared to the "0.5% CMC / 0.5% CFA" group. This suggests that the analgesic composition (Pavlova) at least has the effect of suppressing thermal hyperalgesia.

[0067] [Second Example (Reference Example)] <Preparing the drugs to be used> Diclofenac sodium (DFC, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was prepared by purchasing it from the market. In this example, the DFC was dissolved in physiological saline to a concentration suitable for the dose described below.

[0068] <Verification of the effect of DFC on thermal hyperalgesia> (Creation of arthritis model) Arthritis model mice were prepared in a similar manner to Example 1. Specifically, mice were bred as described in Example 1. Under inhalation anesthesia with isoflurane (Fujifilm Wako Pure Chemical Industries, Ltd.) / oxygen, 50% by mass CFA (Invitrogen) diluted with IFA (Invitrogen) was administered around the right tibial joint to induce arthritis. 15 μL of 50% by mass CFA was administered from the inside and outside of the joint (30 μL in total).

[0069] (evaluation) Three days after the administration of 50% CFA by mass, DFC was intraperitoneally administered to the arthritis model mice at doses of 0.1 mg / kg, 1 mg / kg, or 10 mg / kg. As a control group, saline was intraperitoneally administered to the arthritis model mice at a volume of 10 mL / kg. Seven to eight arthritis model mice were used in each group. The reaction latency of the arthritis model mice was then measured immediately (0 h), 0.5 h, 1 h, 1.5 h, 2 h, 3 h, and 24 h after the administration of DFC or saline using the "Method for Measuring Thermal Hyperalgesia" described in Example 1. A line graph of the results is shown in Figure 1A. Furthermore, the area under the curve (AUC) was calculated from the graph in Figure 1A as an index of the magnitude of the effect, and this is shown in Figure 1B.

[0070] Figure 1A is a line graph showing the relationship between the amount of DFC administered to arthritis model mice and its antihyperalgesic effect. Figure 1B is a bar graph showing the relationship between the area under the curve for the antihyperalgesic effect calculated from the graph in Figure 1A and the dose from immediately after DFC administration to 24 hours later.

[0071] <Consideration> As shown in Figures 1A and 1B, DFC showed a tendency to have a dose-dependent analgesic effect in arthritis model mice (see Figure 1B in particular). However, in arthritis model mice administered DFC intraperitoneally at a dose of 0.1 mg / kg, almost no analgesic effect was observed compared to the control group of mice administered saline (see Figure 1B in particular).

[0072] [Third Example] <Preparation of samples and drugs> Pavlova was prepared in the same manner as in Example 1. In this example, the Pavlova was suspended in a 0.5% by mass CMC (sodium carboxymethylcellulose) solution and used. Furthermore, DFC was prepared in the same manner as in Example 2. In this example, the DFC was dissolved in saline to a concentration suitable for a dose of 0.1 mg / kg. Hereinafter, in this example, such DFC will be referred to as low-dose DFC.

[0073] <Verification of the combined effect of Pavlova and low-dose DFC on thermal hyperalgesia> To verify the combined effect of Pavlova and low-dose DFC on thermal hyperalgesia in arthritis model mice, Pavlova or CMC as a control was orally administered for 28 days to mice maintained in the same manner as in Example 1. The dose of Pavlova itself was 300 mg / kg / day.

[0074] (Creation of arthritis model mice) Arthritis model mice were produced in the same manner as in Example 1. That is, on day 14 after the oral administration of Pavlova or CMC, 30 μL of 50% by mass CFA was administered periarticularly to produce arthritis model mice.

[0075] (evaluation) Three days after administration of 50% by mass CFA, the CMC-administered group of the arthritis model mice was intraperitoneally administered with 10 mL / kg of saline or with 0.1 mg / kg of DFC (low-dose DFC). Furthermore, the Pavlova-administered group of the arthritis model mice was intraperitoneally administered with low-dose DFC. Of the 16 arthritis model mice administered with CMC, 8 were administered with saline and the remaining 8 with low-dose DFC. Of the 8 arthritis model mice administered with Pavlova, low-dose DFC was administered.

[0076] After administration of saline or low-dose DFC to the arthritis model mice, reaction latencies were measured immediately (0 hour), 0.5 hours, 1 hour, 1.5 hours, 2 hours, 3 hours, and 24 hours using the "Method for Measuring Thermal Hyperalgesia" described in Example 1. A line graph of the results is shown in Figure 2A. Furthermore, the area under the curve (AUC) was calculated from the graph in Figure 2A as an index of the magnitude of the effect, and this is shown in Figure 2B.

[0077] Figure 2A is a line graph comparing the antihyperalgesic effects of low-dose DFC alone and low-dose DFC in combination with Pavlova in arthritis model mice. Figure 2B is a bar graph comparing the areas under the curve for the antihyperalgesic effects calculated from the graph in Figure 2A, from immediately after administration of low-dose DFC alone or low-dose DFC in combination with Pavlova, up to 24 hours after administration.

[0078] <Consideration> Figures 2A and 2B suggest that the administration of Pavlova in combination with low-dose DFC, which has almost no analgesic effect, tends to produce an analgesic effect in arthritis model mice (see Figure 2B in particular).

[0079] [Fourth Example] <Sample preparation> Pavlova was prepared in the same manner as in the first example.

[0080] <Preparation of animals> (Creation of arthritis model mice) Arthritis model mice were produced in the same manner as in Example 1. Specifically, Pavlova or CMC (as a control sample) was orally administered to mice maintained in the same manner as in Example 1 for 17 days. The Pavlova dose was 300 mg / kg / day. Furthermore, on the 14th day after oral administration of Pavlova or CMC, 30 μL of 50% by mass CFA was administered periarticularly to produce arthritis model mice. The control group of mice received the same volume (i.e., 30 μL) of saline instead of 50% by mass CFA.

[0081] (Preparation of sectioned spinal cord) Three days after periarticular administration of saline to the control group, saline was administered intraperitoneally at a volume of 10 mL / kg. Three days after administration of 50% by mass CFA to the arthritis model mice, the CMC-administered group received intraperitoneal administration of saline at a volume of 10 mL / kg or intraperitoneal administration of DFC at a dose of 0.1 mg / kg. Furthermore, the Pavlova-administered group of the arthritis model mice received intraperitoneal administration of DFC at a dose of 0.1 mg / kg. Of the 12 arthritis model mice administered with CMC, six received saline and the remaining six received DFC. Of the 6 arthritis model mice administered with Pavlova, six received DFC. The control group consisted of six mice.

[0082] (Immunohistochemical staining) Each mouse was perfused through the left ventricle with phosphate-buffered saline (PBS, pH 7.2) and 4% by weight paraformaldehyde (Invitrogen) diluted with the PBS under isoflurane (Fujifilm Wako Pure Chemical Industries, Ltd.) / oxygen inhalation anesthesia. The spinal cord was then removed and fixed overnight at 4°C in the same 4% by weight paraformaldehyde used for the perfusion. The spinal cord was then cut into 30 μm-thick sections using a vibratome (trade name: "Leica VT1200S", Leica Biosystems).

[0083] The sectioned spinal cords were then blocked by incubation with 10% normal goat serum (NGS, Invitrogen, hereafter referred to as "PBSGT") diluted in 0.3% Triton X-100-containing PBS (PBST) for 1 hour at room temperature. The blocked spinal cords were then incubated overnight at 4°C with a primary antibody (rabbit anti-PAX2 antibody, 1:500, Mercian) diluted in 10% PBSGT. Subsequently, the sections were incubated overnight at 4°C in the dark with a high concentration of secondary antibody (Alexa 488 goat anti-rabbit Fab fragment antibody, 1:80, Jackson Immunoresearch). This simultaneously labeled the primary antibody and masked the antigenic sites.

[0084] Furthermore, after the spinal cord was reacted with mouse anti-NeuN antibody (1:500, manufactured by Merc) and rabbit anti-c-Fos antibody (1:500, manufactured by Cell Signaling Technology), these antibodies were labeled with secondary antibodies (product name: "Alexa 568 goat anti-mouse antibody", 1:400, manufactured by Invitrogen) and secondary antibody (product name: "Alexa 647 goat anti-rabbit Fab fragment antibody", 1:400, manufactured by Invitrogen) under the conditions of 4°C, in the dark, and overnight. For nuclear staining, a 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) solution was used. The stained spinal cord was encapsulated with ProLong (registered trademark) Diamond Antifade Mountant (manufactured by Invitrogen) and stored at 4°C in the dark until observation. The stained spinal cord was visualized as a fluorescence image by using a confocal microscope system (product name: "Nikon C2", manufactured by Nikon Corporation).

[0085] <Examination of the mechanism of analgesic action of Pavlova> To clarify the mechanism of action of Pavlova, changes in neural activity in laminae I-V corresponding to the posterior horn of the spinal cord of mice in each of the above groups were examined. Specifically, neural activity at the above site was analyzed using the change in the number of c-Fos positive cells as an index. More specifically, as described above, neurons (spinal cord) were labeled with Pax2, an inhibitory neuron marker, and NeuN, which is expressed in all neurons. The neural activity of Pax2+ / NeuN+ cells as inhibitory neurons and Pax2- / NeuN+ cells as excitatory neurons was evaluated using the change in the number of c-Fos positive cells as an index. The resulting fluorescence image is shown in Fig. 3A. Furthermore, from the image in Fig. 3A, a bar graph representing the magnitude of neural activity in mice of each group is shown in Fig. 3B.

[0086] Fig. 3A is a fluorescence image evaluating the mechanism of analgesic action of Pavlova by examining the change in the number of c-FOS positive cells in the spinal cord of arthritis model mice. Fig. 3B is a bar graph representing the evaluation results based on the image in Fig. 3A.

[0087] <Consideration> Figures 3A and 3B show that in the group administered DFC and Pavlova, excitatory nerve activity was suppressed to the same level as in the control group in laminae I to V, which correspond to the dorsal horn of the spinal cord (see Figure 3B). Furthermore, the group administered DFC and Pavlova showed a more pronounced tendency for excitatory nerve activity to be suppressed compared to the group administered DFC alone, suggesting that the mechanism of Pavlova's analgesic effect is the suppression of excitatory nerve activity (see Figure 3B).

[0088] Although the embodiments and examples of the present invention have been described above, it is also intended from the beginning that the configurations of the above-described embodiments and examples may be appropriately combined.

[0089] The embodiments and examples disclosed herein should be considered to be illustrative in all respects and not restrictive. The scope of the present invention is defined by the claims, not by the above description, and is intended to include all modifications within the meaning and scope of the claims.

Claims

1. A pain-relieving composition comprising microalgae as an active ingredient.

2. The analgesic composition according to claim 1, wherein the microalgae is classified into the genus Pavlova.

3. 3. The analgesic composition of claim 1 or claim 2, which is an antihyperalgesic or antiallodynic agent.

4. 3. The analgesic composition according to claim 1 or 2, which is a therapeutic agent for arthralgia.

5. The analgesic composition according to claim 1 or 2, which is a c-FOS activity inhibitor.

6. 3. The analgesic composition according to claim 1 or claim 2, which comprises a nonsteroidal anti-inflammatory agent.

7. The analgesic composition according to claim 1 or 2, wherein the microalgae is administered orally to a living body at a daily dose of 5 mg / kg or more and 300 mg / kg or less.

8. The analgesic composition according to claim 7, which is used in combination with a nonsteroidal anti-inflammatory drug.

9. A material for food or drink, comprising the analgesic composition according to claim 1 or 2.

10. The food and beverage material according to claim 9, which is applied to at least one selected from the group consisting of food and beverage products, oral medicines, and animal feed.

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