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WO2009128395A1 - Promoteur de formation osseuse comprenant un composé 2-aminophénol ou un dérivé de celui-ci comme principe actif - Google Patents

Promoteur de formation osseuse comprenant un composé 2-aminophénol ou un dérivé de celui-ci comme principe actif Download PDF

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Publication number
WO2009128395A1
WO2009128395A1 PCT/JP2009/057325 JP2009057325W WO2009128395A1 WO 2009128395 A1 WO2009128395 A1 WO 2009128395A1 JP 2009057325 W JP2009057325 W JP 2009057325W WO 2009128395 A1 WO2009128395 A1 WO 2009128395A1
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Prior art keywords
bone
aminophenoxazin
added
cells
aminophenol
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English (en)
Japanese (ja)
Inventor
正樹 三宅
恵三 河野
千加子 新井
慎平 牛尾
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Hayashibara Seibutsu Kagaku Kenkyujo KK
Original Assignee
Hayashibara Seibutsu Kagaku Kenkyujo KK
Hayashibara Biochemical Laboratories Co Ltd
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Priority to JP2010508187A priority Critical patent/JP5507445B2/ja
Publication of WO2009128395A1 publication Critical patent/WO2009128395A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/136—Amines having aromatic rings, e.g. ketamine, nortriptyline having the amino group directly attached to the aromatic ring, e.g. benzeneamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/538—1,4-Oxazines, e.g. morpholine ortho- or peri-condensed with carbocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention relates to an osteogenesis promoter, and more particularly to an osteogenesis promoter containing 2-aminophenol or a derivative thereof as an active ingredient.
  • the bone tissue in the body is constantly renewed by bone destruction (bone resorption) by osteoclasts and bone formation by osteoblasts. Maintained.
  • bone destruction bone resorption
  • osteoclasts are differentiated monocyte / macrophage progenitor cells, but differentiation into mature osteoclasts is physiologically abbreviated as macrophage-colony stimulating factor (hereinafter “M-CSF”).
  • M-CSF macrophage-colony stimulating factor
  • RNKL Ligand of Receptor Activator of NF- ⁇ B
  • interleukin-1 (hereinafter abbreviated as “IL-1”) produced from synovial cells and T cells activated by inflammation and tumor necrosis factor (hereinafter referred to as “TNF- ⁇ ”). ) Is activated by inflammatory cytokines, and the expression of RANKL, which is a physiological factor essential for differentiation into osteoclasts, is increased, and interleukin-6 (hereinafter, “ Abbreviated as “IL-6”) and nitric oxide (hereinafter abbreviated as “NO”). This production-induced NO promotes cartilage destruction by enhancing apoptosis of chondrocytes or decreasing cartilage matrix synthesis.
  • IL-1 interleukin-1
  • TNF- ⁇ tumor necrosis factor
  • monocytes / macrophages are activated by NO, differentiation into osteoclasts is induced, and bone destruction proceeds.
  • TNF- ⁇ and IL-6 itself induce differentiation of osteoclast precursor cells into osteoclasts, and prostaglandin E 2 (hereinafter referred to as “PGE 2 ”) from osteoblasts induced by these inflammatory cytokines.
  • PGE 2 prostaglandin E 2
  • bone metabolic disorders diseases caused by bone metabolism abnormalities such as osteoporosis and rheumatoid arthritis (hereinafter referred to as “bone metabolic disorders”), it is considered that several factors overlap to form a pathological condition called bone destruction. It has been.
  • estradiol-containing hormone preparations for the treatment of disorders of bone metabolism such as osteoporosis and rheumatoid arthritis, bisphosphonate compounds, calcitonin preparations, active vitamin D 3 preparations, calcium preparations, isoflavone preparations, selective estrogen receptor modulators (SERM preparations) ), Estradiol-containing hormone preparations, and pharmaceuticals such as vitamin K 2 are generally used.
  • active vitamin D 3 derivatives, estradiol derivatives, third-generation bisphosphonate compounds, and the like have been developed with the aim of excellent therapeutic agents with few side effects.
  • hormone preparations may cause side effects. In some cases, the target of use may be limited.
  • 2-aminophenol also known as questionomycin B
  • 2-aminophenol is a compound represented by the following chemical formula 1, and is oxidatively polymerized to give 2-aminophenoxazin-3-one represented by the following chemical formula 2 (also known as QUESTIOMYB). Converts to thiomycin A) relatively easily.
  • 2-Aminophenol or a derivative thereof has long been known as a substance having antibacterial activity, and in particular, 2-aminophenoxazin-3-one is also known as a basic skeleton of actinomycin D, which is a potent anticancer agent. ing. Also, in Motohashi et al., “Medical Research Review”, Vol. 11, pp.
  • 2-aminophenoxazin-3-one and its derivatives have antitumor activity.
  • Shimamoto et al. “Clinical Cancer Research”, Vol. 7, pp. 704 to 708, 2001, one of its derivatives, 2-amino-4,4 ⁇ -dihydro-4 ⁇ , 7-dimethyl-phenoxazin-3-one is disclosed to exhibit cytotoxic activity against various tumor cells.
  • 2-aminophenoxazin-3-one has been disclosed in Japanese Patent Application Laid-Open No. 2004-143101 for the treatment of viral diseases, Japanese Patent Application Laid-Open No. 2005-272334 for the treatment of chlamydiasis, and Japanese Patent Application Laid-Open No.
  • An object of the present invention is to provide an osteogenesis promoter that has fewer side effects than conventional osteogenesis promoters and is highly effective in preventing or improving bone metabolic disorders.
  • 2-aminophenol or a derivative thereof has an action of inducing differentiation of pre-osteoblasts into osteoblasts and promoting calcification. Furthermore, it was discovered that these compounds also have the effect of inhibiting bone destruction by suppressing the induction of osteoclast differentiation from macrophages, and these compounds are effective in the prevention or treatment of diseases of abnormal bone metabolism.
  • the present invention has been completed by finding that it has both a bone formation promoting action and a bone resorption inhibiting action.
  • the present invention solves the above problems by providing an osteogenesis promoter containing 2-aminophenol or a derivative thereof as an active ingredient.
  • an osteogenesis promoter that has fewer side effects than conventional osteogenesis promoters and is excellent in the effect of promoting osteogenesis.
  • the 2-aminophenol referred to in the present invention is a compound represented by the following chemical formula 1, and a commercially available product can be used regardless of its origin and origin as long as the effects of the present invention are exhibited.
  • the derivative of 2-aminophenol as used in the present invention means 2-aminophenoxazin-3-one represented by the following chemical formula 2, which is an oxidized polymer of 2-aminophenol, and a derivative having the basic skeleton as the following. It is as effective as 2-aminophenol.
  • 2-Aminophenoxazin-3-one regardless of its origin or origin, can be produced by appropriate extraction and purification methods from plants and bacteria containing abundant sources, and the precursor 2-aminophenol can be oxidatively polymerized. Can be synthesized.
  • a method of reacting 2-aminophenol with a trivalent iron ion such as potassium ferricyanide described in JP-A-2003-2878, or 2-aminophenol described in JP-A-2-193984 It can be synthesized by a method of reacting with human or bovine hemoglobin.
  • Examples of 2-aminophenoxazin-3-one derivatives used in the present invention include those represented by the following general formula 1 or general formula 2, and salts thereof may also be used. Moreover, as long as the effect of this invention is exhibited, a commercial item can also be used regardless of the origin or origin.
  • those represented by the following general formula 1 include 2-amino-7-hydroxy-phenoxazin-3-one (in the general formula 1, R 5 is a hydroxy group, and R 1 to R 4 are Hydrogen atom), 2-amino-7-methoxy-phenoxazin-3-one (in the following general formula 1, R 5 is a methoxy group, R 1 to R 4 are hydrogen atoms), 2-amino-3,4-dimethoxy- 6,7-dimethoxycarbonyl-phenoxazin-3-one (in the following general formula 1, R 3 and R 4 are methoxy groups, R 6 and R 7 are methoxycarbonyl groups, R 1 , R 2 and R 5 are hydrogen atoms ), 2-acetylamino - phenoxazine-3-one (in the following general formula 1, R 1 is acetyl group, R 2 to R 5 is a hydrogen atom), 2-acetylamino-7-hydroxy - phenoxazine - - one (the)
  • 2-amino-10- (N-4′-aminobutyl) -phenoxazin-3-one (wherein R 1 to R 7 and R 9 in the general formula 2 are A hydrogen atom, R 8 is an aminobutyl group), 2-amino-7-hydroxy-10- (N-4′-aminobutyl) -phenoxazin-3-one (R 1 to R 4 , R 6 of formula 2) R 7 and R 9 are hydrogen atoms, R 5 is a hydroxy group, R 8 is an aminobutyl group), 2-amino-7-methoxy-10- (N-4′-aminobutyl) -phenoxazin-3-one (R 1 to R 4 , R 6 , R 7 and R 9 in the general formula 2 are hydrogen atoms, R 5 is a methoxy group, and R 8 is an aminobutyl group) and salts thereof.
  • sugars may be added to these 2-aminophenoxazin-3-ones and their derivatives, and these compounds by artificial methods to form glycosides, and water-soluble substances such as polyethylene glycol and pullulan. Including those having a binding polymer.
  • R 1 and R 2 are each independently selected from a hydrogen atom (H), a hydroxy group (OH), an acetyl group (COCH 3 ), a hydroxyacetyl group (COCH 2 OH), and R 3 to R 7 are each independently a hydrogen atom (H), a hydroxy group (OH), a methoxy group (OCH 3 ), an ethoxy group (OC 2 H 5 ), a methoxycarbonyl group (COOCH 3 ), an ethoxycarbonyl group (COOC 2). H 5 )
  • R 1 and R 2 are each independently selected from a hydrogen atom (H), a hydroxy group (OH), an acetyl group (COCH 3 ), a hydroxyacetyl group (COCH 2 OH), and R 3 to R 7 and R 9 are each independently a hydrogen atom (H), a hydroxy group (OH), a methoxy group (OCH 3 ), an ethoxy group (OC 2 H 5 ), a methoxycarbonyl group (COOCH 3 ), or an ethoxycarbonyl group.
  • R 8 is an amino propyl group (CH 2 CH 2 CH 2 NH 2), is selected from amino-butyl group (CH 2 CH 2 CH 2 CH 2 NH 2).
  • the disease of abnormal bone metabolism as used in the present invention refers to a disease in which bone balance caused by osteoclasts and bone formation by osteoblasts is lost, and the bone mass is reduced compared to healthy subjects. Including those with reserves. Specific examples include osteoporosis, rheumatoid arthritis, rheumatoid spondylitis, Paget's disease of bone, osteoarthritis, gouty arthritis, gingivitis, periodontal disease, and their reserves.
  • the bone formation promoting action referred to in the present invention means an action in which bone formation by osteoblasts is promoted to increase bone mass, and includes the case where bone loss is stopped or improved. Specifically, it means an action of promoting bone formation by promoting calcium deposition by increasing osteoblast activity, promoting differentiation, or increasing the number of cells.
  • the osteoclast-inhibiting action referred to in the present invention means an action to suppress bone loss due to increased bone destruction (bone resorption) in osteoclasts, and induces osteoclast differentiation from macrophages by RANKL stimulation.
  • Inhibition inhibition of IL-6 production from osteoblasts, inhibition of PGE 2 synthesis by inhibiting the activity of cyclooxygenase (hereinafter abbreviated as “COX”), and inducible monoxide
  • COX cyclooxygenase
  • inducible monoxide Includes the action of inhibiting the synthesis of macrophages into osteoclasts based on the action of inhibiting the synthesis of nitrogen by macrophages by suppressing the production of nitrogen synthase (hereinafter abbreviated as “iNOS”).
  • the NO synthesis inhibitory activity referred to in the present invention is exhibited by quantitatively decreasing iNOS in a cell, and the compound of the present invention is added to a cell having NO synthesis activity, for example, a macrophage cell. It can be examined by measuring the amount of NO produced by the action of iNOS.
  • macrophage cells mouse-derived cell line RAW264.7 cells or those collected from experimental animals such as mice can be used. The amount of NO produced can be measured by a conventional Griess method.
  • Cyclooxygenase (COX) inhibitory activity in the present invention in the COX-1 or COX-2 and arachidonic acid presence, by adding a compound of the invention as the test sample, measuring the amount of generated PGE 2, This can be confirmed by comparison with a control sample.
  • the 50% inhibitory concentration (IC 50 value) represents the concentration of the test sample required to inhibit PGE 2 synthesis by 50% compared to an uninhibited control sample. Further, by adding a test sample to cells having cyclooxygenase activity can also be confirmed by measuring the production of PGE 2.
  • the COX-1 / COX-2 ratio a ratio of an IC 50 value with respect to an IC 50 value and COX-2 relative to COX-1, as this value is larger, relative to COX-2 All selective inhibitors.
  • 2-aminophenol or a derivative thereof used in the present invention has a bone formation promoting action.
  • these compounds produce physiologically active substances involved in enhancing bone destruction, such as NO synthesis inhibitory action, PGE 2 production inhibitory action, cyclooxygenase activity inhibitory action, and IL-6 and other inflammatory cytokine production inhibitory actions. It has the effect
  • 2-aminophenol or a derivative thereof also has an action of enhancing the production of collagen constituting the matrix of bone and cartilage, foods, cosmetics, quasi-drugs intended for osteogenesis promoters, It has a wide variety of uses as pharmaceuticals.
  • the osteogenesis promoter of the present invention can be taken orally or transgastrically or enterally by a catheter or suppository.
  • the content of 2-aminophenol or its derivative as an active ingredient contained in the osteogenesis promoter is based on the total mass of the osteogenesis promoter, Usually, it is 0.01 to 100% by mass, preferably 0.1 to 50% by mass, more preferably 0.5 to 10% by mass, and the amount used is appropriately determined according to the degree of symptoms of bone metabolic disorder. It may be determined, may be taken every other day, may be taken every day, or may be taken once to several times a day.
  • the amount of intake is usually 0.05 to 100 mg / kg / body weight, preferably 0.5 to 50 mg / kg / body weight, more preferably 2.5 To 25 mg / kg ⁇ body weight.
  • osteogenesis promoter of the present invention other substances having an osteogenesis promoting action or a bone destruction inhibiting action can be appropriately blended as long as the action and effects of 2-aminophenol or a derivative thereof are not inhibited.
  • bisphosphonate compound, calcitonin preparation, active vitamin D 3 preparation, hormone preparation containing estradiol, SERM preparation, isoflavone preparation, vitamin K 2 , mineral (calcium, magnesium, etc.) preparation, trehalose examples thereof include substances having an action of inhibiting bone destruction and / or promoting bone formation, such as lactosucrose.
  • bisphosphonate compounds are particularly preferable because they enhance the osteogenesis promoting action of the osteogenesis promoter of the present invention synergistically or additively.
  • various optional components that are usually used in foods, quasi drugs, pharmaceuticals, and the like can be appropriately blended as necessary.
  • water, ethanol, glycerin, moisturizer, oil component, emulsifier, emulsion stabilizer, thickener, antioxidant, preservative, pH adjuster, fragrance, physiologically active substance, medicinal component, animal / plant extract, etc. can be mentioned.
  • 2-aminophenol or a derivative thereof used in the present invention is very hydrophobic, when used in the form of a solution, after previously mixing with oil, lecithin, polyethylene glycol, saccharides, etc., What is necessary is just to mix with a small amount or an appropriate amount of water, and to use as an emulsification thru
  • it may be in the form of micelles encapsulated in liposomes or dissolved in an organic solvent such as acetone together with a hydrophilic-hydrophobic polymer and then mixed with water and encapsulated in the polymer.
  • the dosage form of the osteogenesis promoter of the present invention may be in a form suitable for oral ingestion or gastric / enteral administration, for example, liquid, tablet, troche, pill, aqueous suspension, oil suspension, Examples include dispersible powders or granules, emulsions, hard capsules, soft capsules, syrups, and elixirs.
  • oral, gastric and enteral administration topical administration such as subcutaneous, intradermal, intramuscular, intravascular, intraperitoneal, etc., or respiratory tract, pulmonary administration by inhalation, spraying, etc. Can be administered parenterally, and a dosage form suitable for these is also optional.
  • the dosage in the case of parenteral administration may be determined as appropriate according to the degree of the symptom of abnormal bone metabolism, and may be administered every other day or every day, once to several times a day. It may be administered separately. In this case, the dose is usually 0.0005 to 20 mg / kg / body weight, preferably 0.005 to 5 mg / kg / body weight, more preferably 2-aminophenol or a derivative thereof. 0.05 to 2.5 mg / kg body weight.
  • the osteogenesis promoter of the present invention is an effective ingredient other than the active ingredient of the osteogenesis promoter of the present invention for the purpose of improving or suppressing pain, inflammation and the like accompanying abnormal bone metabolism within the range where the desired effect is obtained.
  • analgesics such as acetaminophen and phenacetin
  • potentiators of analgesic action of analgesics such as caffeine, H2-antagonists, aluminum hydroxide, magnesium hydroxide, simethicone, diuretics, analgesic anti A histamine drug, a non-analgesic antihistamine, etc. are mentioned, What is necessary is just to use together with these 1 type, or 2 or more types.
  • the bone formation-promoting agent of the present invention can be used, for example, for bone metabolism abnormal diseases accompanied by bone destruction such as rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis, gingivitis, periodontal disease, osteoporosis, especially bone destruction It can be used as a prophylactic agent, ameliorating agent, or therapeutic agent for bone metabolic disorders associated with inflammatory reactions.
  • the osteogenesis promoter of the present invention has an anti-inflammatory action in addition to the osteogenesis promoting action, it can also be used in applications such as inflammatory diseases, analgesics, and antipyretics.
  • the osteogenesis promoter of the present invention can also be used as a therapeutic agent for various cancers in which COX-2 is overexpressed.
  • the osteogenesis promoter of the present invention is administered in the form of a dosage unit preparation containing a pharmaceutically acceptable base material, additives and excipients. Moreover, it can be applied not only to the treatment or prevention of diseases of abnormal bone metabolism in humans, but also to livestock such as mice, rats, horses, cows, sheep, pigs, dogs, cats, chickens, poultry and pets.
  • the solid obtained by drying the reaction solution under reduced pressure was dissolved in 450 ml of methanol, centrifuged to remove undissolved solids, and then dried again under reduced pressure.
  • the obtained solid was dissolved in 200 ml of ethyl acetate, 200 ml of 0.005N hydrochloric acid aqueous solution was added and stirred, and then the ethyl acetate layer was recovered with a separatory funnel. After repeating this operation three times, the obtained ethyl acetate layer was concentrated under reduced pressure until the liquid volume became 15 ml, and this was followed by a silica gel chromatography column (Wako Pure Chemical Industries, Ltd.
  • ⁇ Experiment 2 Effect on mouse preosteoblast cell line MC3T3-E1> ⁇ -MEM medium (Gibco Invitrogen) containing 10 volume% fetal calf serum (sold by JR Scientific, hereinafter abbreviated as “FCS”) in a 24-well plate (sold by Becton Dickinson).
  • FCS fetal calf serum
  • FCS-added ⁇ -MEM medium 10 volume% fetal calf serum
  • the powdery 2-aminophenoxazin-3-one prepared in Experiment 1 as a test substance or 2-aminophenol as its starting material is dissolved in ethanol. Used. As a positive control, it used as a therapeutic agent for osteoporosis, and, using vitamin K 2 having a function of promoting the deposition of calcium into the bone (Ca).
  • a differentiation-inducing medium (10 vol% FCS-added ⁇ -MEM medium + 10 mM ⁇ -glycerophosphate (sold by Sigma) +10 mM HEPES (DOJINDO Wako Pure Chemicals) supplemented with this test substance or positive control to the concentration shown in Table 1 Kogyo Co., Ltd.) was added at 2 ml / well. Furthermore, as a negative control, a differentiation induction medium supplemented with ethanol used as a solvent so that the final concentration was 0.1% by mass was added and cultured.
  • ALP Alkaline phosphatase activity
  • AP is 2-aminophenol in Table 1
  • QA means 2-amino-phenoxazine-3-one (QUEStionable thio mycin A)
  • VK means vitamin K 2.
  • culture is performed once every 3 or 4 days on the same day for all wells in the differentiation induction medium containing the test substance or positive control at the same concentration as the first addition or in the negative control differentiation induction medium.
  • the measured value when the test substance or the positive control is added is divided by the measured value of the negative control and multiplied by 100 to obtain a relative value with the measured value of the negative control as 100%, and the rate of increase in ALP activity (% ) are also shown in Table 1.
  • Method for measuring osteocalcin> On the ninth day from the start of the culture, the cell culture supernatant was collected and measured using a commercially available EIA kit for measuring mouse osteocalcin (available from Biomedical Technologies Inc., MA. USA).
  • the measured value when the test substance or positive control is added is divided by the measured value of the negative control and multiplied by 100 to obtain a relative value with the measured value of the negative control as 100%, and the increase rate of the secretion amount of osteocalcin It is also shown in Table 1 as (%).
  • Dulbecco's PBS (-) available from Nissui Pharmaceutical Co., Ltd. was added to each well from which the culture supernatant had been removed to wash the cells.
  • ⁇ Cell viability> The cells are stained with an alamar blue dye (trade name “Alamar Blue Assay”, sold by Trek Diagnostics Systems, Inc.) by a conventional method, and measured with a fluorescent plate reader (sold by Molecular Devices), The measured value when the test substance or the positive control is added is divided by the measured value of the negative control and multiplied by 100 to obtain a relative value with the measured value of the negative control as 100%, and the cell viability (% ) Are also shown in Table 1.
  • alamar blue dye trade name “Alamar Blue Assay”, sold by Trek Diagnostics Systems, Inc.
  • a fluorescent plate reader sold by Molecular Devices
  • the ALP activity and the amount of calcium deposition of MC3T3-E1 cells cultured with 2-aminophenol (AP) or 2-aminophenoxazin-3-one (QA) added to the medium As compared with the case where only ethanol used as a culture was added (negative control), it increased depending on the added concentration.
  • the amount of osteocalcin secreted is determined when 2-aminophenol (AP) or 2-aminophenoxazin-3-one (QA) is added to the medium at 1 or 2 ⁇ M, and only ethanol used as a solvent is added. Increased compared to (negative control).
  • Vitamin K 2 was used as a positive control also, in the case of adding only ethanol to the medium than the (negative control), ALP activity and calcium deposition amount, but rose in dependence on the added concentration of 10 ⁇ M In addition, ALP activity was suppressed reversibly. Moreover, secretion of osteocalcin, the addition of vitamin K 2 so as to be 10 [mu] M, compared to negative controls, increase was observed. When compared with the rate of increase in the amount of calcium deposited, 2-aminophenoxazin-3-one showed a stronger calcium deposition enhancing action than vitamin K 2 used as a positive control.
  • 2-aminophenol and its derivatives have the effect of inducing differentiation of preosteoblasts into osteoblasts and promoting calcification in the cells, and these substances can be used in chronic joints. It shows that it can be used as an osteogenesis promoter or calcification promoter for bone metabolism disorders such as rheumatism, osteoporosis, periodontal disease, and the reserves. It also shows that 2-aminophenoxazin-3-one (QA) is superior to 2-aminophenol (AP) in terms of the strength of promoting bone formation. Furthermore, 2-aminophenol and 2-aminophenoxazin-3-one used in this experiment did not affect cell viability at any concentration used in the experiment. The cytotoxicity of aminophenol and 2-aminophenoxazin-3-one was judged to be weak.
  • MC3T3-E1 cells were seeded in a 24-well plate, and one of the differentiation induction media in which the test substance was dissolved was added at 2 ml / well to the semi-confined cells. Once every 3 or 4 days, all wells were cultured on the same day for 20 days while changing the medium with a differentiation-inducing medium containing the same kind and concentration of the test substance as the first added one.
  • the amount of calcium deposited in the cells on the 20th day from the start of the culture was measured, and the amount of calcium deposited in the cells cultured in the medium supplemented with the derivative of 2-aminophenoxazin-3-one was Divide by the amount of calcium deposited in cells cultured in a medium supplemented with phenoxazin-3-one and multiply by 100 to give a relative value with the amount of calcium deposited when 2-aminophenoxazine-3-one added as 100%. Obtained and shown in Table 2 as the Ca (calcium) deposition rate (%).
  • the cell viability (%) is obtained by the same method as in Experiment 2 and is also shown in Table 2.
  • the cell survival rate (%) was expressed as a relative value with the survival rate of cells cultured in a medium supplemented with 2-aminophenoxazin-3-one being 100%. The test was conducted in 3 wells per sample.
  • test substance 2-aminophenoxazin-3-one was added to the concentration shown in Table 3, and lipolipin as an inducer of NO and PGE 2 production was added.
  • Polysaccharide was added at 2 ⁇ g / ml and IFN- ⁇ at 10 IU / ml, and RPMI-1640 medium containing 10 vol% FCS was added to make the volume 200 ⁇ l. After culturing this at 37 ° C.
  • osteoprogenitor cell line MC3T3-E1 cells from C57BL / 6 mice expressing it as a RANKL ligand. It was done as follows. Specifically, bone marrow cells were collected from the femurs of C57BL / 6 mice (female, 7 weeks old, sold by Charles River Japan, Inc.) by a conventional method, washed twice with ⁇ -MEM medium, and added with 10% by volume FCS. The suspension was suspended in ⁇ -MEM medium and adjusted to a concentration of 4 ⁇ 10 6 cells / ml.
  • MC3T3-E1 cells are suspended in a 50 ⁇ g / ml mitomycin C (hereinafter sometimes abbreviated as “MMC”) solution, kept in a water bath at 37 ° C. for 30 minutes, and then placed in ⁇ -MEM medium. The cells were washed 3 times and suspended in ⁇ -MEM medium supplemented with 10% by volume FCS to a concentration of 4 ⁇ 10 5 cells / ml. Add 100 ⁇ l / well of this bone marrow cell suspension and 50 ⁇ l / well of MMC-treated MC3T3-E1 cells to a 48-well plate (Becton Dickinson, trade name “Multiwell TM 48-well”).
  • MMC mitomycin C
  • Recombinant human M-CSF 120 ng / ml, sold by Pepro Tech was added at 50 ⁇ l / well. Further, 2-aminophenoxazin-3-one of the test substance dissolved in 10% by volume FCS-added ⁇ -MEM medium was added to this well at 50 ⁇ l / well at 37 ° C. The culture was started. As a negative control, the cells were cultured for 9 days in a medium supplemented only with M-CSF.
  • alendonate alendronate, sold by LKT Laboratories
  • raloxifene hydrochloride raloxifene, sold by LKT Laboratories
  • Table 5 shows the results of the test conducted after incubation.
  • the culture is carried out once every 3 or 4 days on the same day for all wells, a medium containing the same concentration of M-CSF as that initially added and the test substance or positive control, or M-CSF.
  • the culture was performed for 9 days while the medium was changed with a negative control medium containing.
  • the test was performed in 3 wells per sample.
  • mice bone marrow cells were treated with TRAP in the presence of M3CSF and RANKL ligand-expressing MC3T3-E1 cells only (see 2-aminophenoxazin-3-one concentration of 0 ⁇ M). Differentiated into positive cells.
  • 2-aminophenoxazin-3-one was added and cultured in the presence of M-CSF and MC3T3-E1 cells, it was the same as in the case of alendonate or raloxifene hydrochloride added as a positive control.
  • mouse bone marrow cells produced IL-6 in the presence of M-CSF and MC3T3-E1 cells alone (2-aminophenoxazin-3-one concentration 0 ⁇ M), whereas When phenoxazin-3-one was added, its production was suppressed.
  • ⁇ Experiment 7 Effect of mouse spleen macrophages on differentiation into osteoclasts in the presence of M-CSF and soluble RANKL> Since it was confirmed in Experiment 6 that osteoclasts can be induced from bone marrow cells by co-culture with MC3T3-E1 cells expressing RANKL ligand, soluble in the presence of M-CSF using mouse spleen macrophages The following tests were conducted to examine the effect of 2-aminophenoxazin-3-one on the differentiation of macrophages into osteoclasts by RANKL (soluble RANKL is abbreviated as “sRANKL”). .
  • spleen cells were collected from BALB / c mice (female, 7 weeks old, sold by Charles River Japan, Inc.) by a conventional method, washed twice with ⁇ -MEM medium (MP Biomedical), and then a 48-well plate. (Becton Dickinson, trade name “Multiwell TM 48 well”) was suspended in ⁇ -MEM medium supplemented with 10% by volume FCS and seeded at 8 ⁇ 10 4 cells / well. Macrophages attached to the plate after 2 hours incubation at 37 ° C. with 5% by volume CO 2 were washed twice with ⁇ -MEM medium, and then fresh 10% by volume FCS-added ⁇ -MEM medium was 200 ⁇ l / well.
  • ⁇ -MEM medium MP Biomedical
  • test substance aminophenoxazin-3-one
  • the differentiation-inducing medium so as to have the concentrations shown in Table 6, and the culture was continued.
  • the cells in each well were stained using a TRAP staining kit that selectively stains osteoclasts as in Experiment 5, and stained under a microscope.
  • the number of TRAP positive cells was counted.
  • the results are also shown in Table 6.
  • bone marrow cells were similarly cultured in a medium supplemented with M-CSF alone and a medium supplemented with M-CSF and sRANKL.
  • risedronate (Risedronate, sold by LKT Laboratories) used as a therapeutic agent for osteoporosis was added instead of aminophenoxazin-3-one, and cultured in the same manner.
  • culture is performed once every 3 or 4 days on the same day for all wells, and the medium is replaced with a medium containing the test substance or positive control at the same concentration as the first added or a negative control medium. The culture was continued for 14 days. The test was performed in 3 wells per sample.
  • mouse macrophages do not differentiate into TRAP-positive cells in the presence of M-CSF alone, but coexist with M-CSF and sRANKL (2-aminophenoxazine-3-one).
  • the concentration of the cells was differentiated into TRAP positive cells.
  • 2-aminophenoxazin-3-one was added together with M-CSF and sRANKL
  • M-CSF and sRANKL were added in the same manner as when risedronate used as a positive control was added and cultured.
  • the increase in the number of TRAP positive cells depending on the concentration was suppressed.
  • ⁇ Experiment 8 Effect of TNF- ⁇ or IL-1 on osteoblast activation> Since rheumatoid arthritis and periodontal disease are usually accompanied by inflammation, inflammatory cytokines (IL-1, TNF- ⁇ ) produced from activated synovial cells and T cells are secreted at the inflammation site. Thus, osteoblasts are activated, the expression of RANKL, which is a physiological factor essential for differentiation into osteoclasts, is increased, and the production of IL-6 and NO is increased. This production-induced NO enhances the apoptosis of chondrocytes or decreases the synthesis of cartilage matrix, and therefore promotes the destruction of cartilage at the inflammatory site.
  • RANKL which is a physiological factor essential for differentiation into osteoclasts
  • monocytes / macrophages are activated by NO, differentiation into osteoclasts is induced, and bone destruction (bone resorption) proceeds. Furthermore, there is a direct differentiation induction pathway from osteoclast precursor cells to osteoclasts by TNF- ⁇ itself. Thus, in rheumatoid arthritis, it is considered that several pathways overlap to form a pathological condition of bone destruction. Therefore, a test for examining the effect of 2-aminophenoxazin-3-one on the activation of osteoblasts by TNF- ⁇ or IL-1 was conducted using MC3T3-E1 cells as osteoblasts as follows. It was done.
  • MC3T3-E1 cells prepared in 10% by volume FCS-added ⁇ -MEM medium to a concentration of 5 ⁇ 10 5 cells / ml were prepared in a 96-well microplate (available from Becton Dickinson, trade name “Microtest TM 96 ”) at 50 ⁇ l / well.
  • 2-aminophenoxazin-3-one diluted in the same medium was added at 50 ⁇ l / well so that the concentrations shown in Table 7 or 8 were obtained.
  • Cell proliferation was determined by adding 100 ⁇ l / well of an Alamar Blue stock solution diluted 4-fold in the same medium and incubating at 5% by volume CO 2 at 37 ° C. for 2 to 3 hours, followed by absorbance at 544/590 nm. Was measured. The test was performed in 3 wells per sample.
  • the measurement results of NO production and IL-6 production in the presence of recombinant human IL-1 ⁇ are shown in Table 7, and the measurement results in the presence of recombinant human TNF- ⁇ are also shown in Table 8.
  • MC3T3-E1 cells were present only in the presence of recombinant human IL-1 ⁇ or recombinant human TNF- ⁇ (see 2-aminophenoxazin-3-one concentration of 0 ⁇ M). NO and IL-6 production was observed. When cultured by adding 2-aminophenoxazin-3-one together with recombinant human IL-1 ⁇ , NO and IL are compared to when cultured with only recombinant human IL-1 ⁇ . The production of -6 was strongly suppressed.
  • ⁇ Experiment 9 Effect of mouse spleen macrophages on differentiation into osteoclasts in the presence of M-CSF and TNF- ⁇ >
  • Induction of differentiation from macrophages into osteoclasts is not limited to the pathway by osteoblasts, but especially in diseases such as rheumatoid arthritis and periodontal disease, chronically produced TNF- ⁇ and IL-1 are M- It has been revealed that there is also a pathway that directly acts on macrophages in the presence of CSF to induce differentiation into osteoclasts. Therefore, a test for examining the effect of 2-aminophenoxazin-3-one on differentiation induction from macrophages into osteoclasts in the presence of TNF- ⁇ and M-CSF was performed as follows.
  • spleen cells were collected from C57BL / 6 mice (female, 7 weeks old, sold by Charles River Japan, Inc.) by a conventional method, washed twice with ⁇ -MEM medium, and then added to ⁇ -MEM medium supplemented with 10% FCS. The suspension was suspended and seeded in a 48-well plate (trade name “Multiwell TM 48-well”, sold by Becton Dickinson) at 6 ⁇ 10 4 cells / well. Mouse spleen macrophages that had been incubated for 2 hours at 37 ° C.
  • FCS-added ⁇ -MEM medium was 200 ⁇ l / Wells and recombinant human M-CSF (60 ng / ml, sold by Pepro Tech) were added at 100 ⁇ l / well, followed by culturing at 37 ° C. for 5 days. The culture supernatant of each well was completely removed by aspiration, and 10 volume% FCS-added ⁇ -MEM medium containing 20 ng / ml human recombinant M-CSF and 20 ng / ml recombinant human TNF- ⁇ was added. Added at 200 ⁇ l / well.
  • 2-aminophenoxazin-3-one as a test substance was added to the medium so as to have a concentration shown in Table 9, and the culture was continued for 14 days. Culturing was continued for 14 days in a medium supplemented with M-CSF and TNF- ⁇ as a negative control. After removing the culture supernatant, as in Experiment 6, the cells in each well were stained using a TRAP staining kit that selectively stains osteoclasts, and the number of stained TRAP-positive cells was counted under a microscope. The results are also shown in Table 9. As a positive control, risedronate was added instead of aminophenoxazin-3-one, and culture was performed in the same manner.
  • culture is performed once every 3 or 4 days on the same day for all wells, and the medium is replaced with a medium containing the test substance or positive control at the same concentration as the first added or a negative control medium.
  • the culture was continued for 14 days.
  • the test was performed in 3 wells per sample.
  • 2-aminophenoxazin-3-one was added together with M-CSF and TNF- ⁇
  • M-CSF was added in the same manner as when risedronate used as a positive control was added and cultured.
  • the increase in the number of TRAP-positive cells depending on the concentration was observed.
  • 2-aminophenoxazin-3-one like risedronate used as a positive control, can suppress the differentiation of macrophages into osteoclasts induced by inflammatory cytokines such as TNF- ⁇ . ing.
  • 2-aminophenoxazine is also used in a system in which RAW-264.7 cells of the macrophage cell line are stimulated with recombinant human TNF- ⁇ to induce differentiation into osteoclasts.
  • -3-one was found to suppress the increase in the number of TRAP-positive cells depending on its concentration.
  • mice Forty ddy mice (8 weeks old, male, sold by Nippon Charles River Co., Ltd.) were randomly divided into 10 groups of 4 groups, and 10 groups of 3 ovaries were extracted. The remaining 10 animals per group, as control 1, performed only laparotomy without removing the ovaries. From the day after the ovary was removed, 10 animals in each of the 2 groups were treated with 2-aminophenoxazin-3-one at a dose of either 3 mg / kg ⁇ body weight or 10 mg / kg ⁇ body weight using a gastric sonde, It was administered 5 times / week and reared for 4 weeks.
  • control 2 The remaining 10 animals (control 2) from which the ovaries had been removed and 10 animals in control 1 group were given an aqueous solution containing 1.5% of 1N hydrochloric acid and 0.5% by mass of carboxymethylcellulose, respectively. Used and dosed 5 times / week and raised for 4 weeks. 29 days after the ovariectomy operation, blood was collected from the abdominal vena cava under ether anesthesia, and then the mouse was dissected and the right femur was removed. The femur was immersed in 70% by volume ethanol overnight, and then tissues such as attached muscle were removed and dried overnight in an oven at 110 ° C. The dried femur was put into a crucible, heated in an oven at 950 ° C.
  • femur bone ash mass, Ca, Mg and P as compared to mice (control 2) administered with an aqueous solution containing 5% and carboxymethylcellulose 0.5% by weight with a gastric sonde for 4 weeks. Any decrease in the content of was suppressed.
  • the amount of osteocalcin in the serum is higher than that of a laparotomy-only mouse (control 1) in which the ovaries are not removed, and after removing the ovaries, an aqueous solution containing 1.5% by mass of 1N hydrochloric acid and 0.5% by mass of carboxymethylcellulose.
  • mice administered with a gastric tube for 4 weeks (Control 2), the concentration decreased.
  • osteocalcin binds to calcium efficiently. It shows that bone formation is promoted by being taken up by cells.
  • the body weight of the mice was measured. As a result, there was no difference between the four groups in the increase or decrease in the body weight throughout the test period, and 2-aminophenoxazin-3-one was toxic even if taken orally. It was judged to be a low and safe compound.
  • the cell name “NHDF”) was suspended at 5.0 ⁇ 10 5 cells / ml, and 50 ⁇ l / well was added to a 96-well microplate (available from Becton Dickinson). 2 and cultured for 1 day. The culture supernatant of each well was removed, and 10 volume% FC-added D-MEM containing 100 ⁇ M ascorbic acid 2-glucoside (trade name “AA2G”, sold by Hayashibara Biochemical Co., Ltd.) was added at 100 ⁇ l / well. did.
  • a solution prepared by dissolving 2-aminophenoxazin-3-one prepared in Experiment 1 as a test substance in DMSO to 4 mM was diluted with 10% by volume FCS-added D-MEM, and added at 100 ⁇ l / well. The final concentration of each well was adjusted to the concentration shown in Table 11. After culturing at 37 ° C. under 5% by volume CO 2 for 3 days, the medium was replaced with the same composition, and the culturing was further continued for 3 days.
  • a 1 mM acetic acid solution of pepsin (sold by Sigma) (pepsin concentration 1 mg / ml) was added at 50 ⁇ l / well at room temperature.
  • the pepsin digest was obtained by shaking for 4 hours.
  • the digested product is pipetted and collected in a 1.5 ml centrifuge tube, and 200 ⁇ l / tube of a color developing reagent of a commercially available collagen quantification kit (trade name “Sircol Collagen Assay Kit”) is added.
  • the mixture was mixed by inverting at room temperature for 30 minutes.
  • the tube for centrifugation was centrifuged as it was at 4 ° C. for 10 minutes (15000 rpm), the supernatant was removed, 50 ⁇ l of 1N NaOH was added to dissolve the precipitate, and the absorbance at 560/650 nm was measured.
  • a known amount of collagen as a standard product was appropriately diluted, digested with pepsin in the same manner, similarly treated with a coloring reagent, and the absorbance was measured to prepare a standard curve. The results of calculating the amount of collagen in NHDF cells based on this standard curve are also shown in Table 11.
  • ⁇ Bone formation promoter 2-Aminophenoxazin-3-one synthesized by the method described in Experiment 1 with respect to 75 parts by mass of anhydrous crystalline ⁇ -maltose powder (trade name “FINE TOSE”, sold by Hayashibara Corporation), or synthesis thereof 2-aminophenol used as a raw material was added in a proportion of 25 parts by mass and mixed uniformly to prepare a powdery osteogenesis promoter.
  • This product can be used as it is, tableted, encapsulated, or added to a soft drink or the like.
  • This product is used as an osteogenesis promoter, osteoporosis, rheumatoid arthritis, rheumatoid spondylitis, Paget's disease, osteoarthritis, gouty arthritis, gingivitis, periodontal disease and other bone metabolic disorders and its It is suitable for suppressing the decrease in bone mass of the reserve army or promoting the increase in bone mass.
  • ⁇ Bone formation promoter> instead of 25 parts by mass of 2-aminophenoxazin-3-one used in Example 1, the derivative of 2-aminophenoxazin-3-one synthesized in Experiment 3 was 2-amino-7-hydroxy-phenoxazine- 3-one, 2-amino-7-methoxy-phenoxazin-3-one, 2-amino-3,4 dimethoxy-6,7-dimethoxycarbonyl-phenoxazin-3-one, 2- (N-hydroxy) acetyl Powdered in the same manner as in Example 1 except that 25 parts by mass of amino-phenoxazin-3-one or 2-amino-10- (N-4'-aminobutyl) -phenoxazin-3-one was added A bone formation promoter was prepared.
  • This product can be used as it is, tableted, encapsulated, or added to a soft drink or the like.
  • This product is used as an osteogenesis promoter, osteoporosis, rheumatoid arthritis, rheumatoid spondylitis, Paget's disease, osteoarthritis, gouty arthritis, gingivitis, periodontal disease and other bone metabolic disorders and its It is suitable for suppressing the decrease in bone mass of the reserve army or promoting the increase in bone mass.
  • ⁇ Bone formation promoter 1 part by weight of sugar ester is added to 99 parts by weight of any of the powdered osteogenesis promoters prepared in Example 1 or 2, mixed homogeneously, and compressed into tablets by a conventional method.
  • a tablet containing about 50 mg of 2-aminophenoxazin-3-one or a derivative thereof or 50 mg of 2-aminophenol was obtained.
  • This product is used as an osteogenesis promoter, osteoporosis, rheumatoid arthritis, rheumatoid spondylitis, Paget's disease, osteoarthritis, gouty arthritis, gingivitis, periodontal disease and other bone metabolic disorders and its It is suitable for suppressing the decrease in bone mass of the reserve army or promoting the increase in bone mass.
  • this product since this product has COX inhibitory activity and nitric oxide synthesis inhibitory activity, it can be administered orally to treat or prevent various inflammatory diseases such as rheumatism and gastritis, cancer, Alzheimer's disease, and analgesics. It can also be used as an antipyretic.
  • ⁇ Bone formation promoter> The following ingredients were blended to obtain a powdery osteogenesis promoter. 41 parts by weight of powdered bone formation promoter prepared in Example 1 or 2 Lactosucrose (powder, lactosucrose purity 90%) 5 parts by weight powdered rice 41 parts by weight glycosylated ascorbic acid (Hayashibara Co., Ltd.) Commercial sales, 1 part by mass, “Asco Fresh”) Kotarahim extract powder 0.2 parts by weight mineral yeast (zinc, selenium, chromium, copper, manganese, 1 part by weight molybdenum and iodine) Carnitine Appropriate vitamin A Appropriate vitamin D Appropriate amount Thiamine hydrochloride Appropriate amount Riboflavin Appropriate amount Pyridoxine hydrochloride Appropriate amount Cyanocobalamin Appropriate amount Choline hydrogen tartrate Appropriate nicotinic acid amide Appropriate amount Tocopherol acetate Ap
  • this product can also be used for the prevention and treatment of all life-week diseases as an inhibitor of blood sugar elevation, an absorption inhibitor of carbohydrates and lipids, and an agent for improving metabolism of sugars and lipids.
  • ⁇ Bone formation promoter> Dentifrice was manufactured with the following formulation. Insoluble sodium metaphosphate 26.0% by mass Glycerin 25.0% by mass Dicalcium phosphate 15.0% by mass Sodium lauryl sulfate 1.5% by mass Tragacanth gum 1.4% by mass Fragrance 1.0% by mass 2-Aminophenoxazine produced in Experiment 1 0.1% by mass 3-one or 2-aminophenol (sold by Wako Pure Chemical Industries, Ltd.) Saccharin 0.1% by mass Copper chlorophyllin sodium 1.0% by mass 28.9% by mass of water
  • This product is used as an osteogenesis promoter, osteoporosis, rheumatoid arthritis, rheumatoid spondylitis, Paget's disease, osteoarthritis, gouty arthritis, gingivitis, periodontal disease and other bone metabolic disorders and its It is suitable for suppressing the decrease in bone mass of the reserve army or promoting the increase in bone mass.
  • ⁇ Bone formation promoter 20 g of vegetable oil and 0.5 g of 2-aminophenoxazin-3-one or 2-aminophenol were mixed and mixed under heating. To this mixed solution, 0.6 g of purified egg yolk lecithin and 5.5 g of glycerin were added and dissolved by vigorous stirring while warming, and then an appropriate amount of distilled water was added and stirred with a Polytron homogenizer to prepare an emulsion. .
  • This emulsified liquid is further emulsified with a microfluidizer, and then distilled water is added to the emulsified liquid to make 100 ml, followed by filtration with a filter having a pore size of 0.45 ⁇ m, so that 2-aminophenoxazin-3-one or 2-amino An injection in which fat particles containing about 400 ⁇ g / ml of phenol were dispersed was prepared.
  • This product is used as an osteogenesis promoter, osteoporosis, rheumatoid arthritis, rheumatoid spondylitis, Paget's disease, osteoarthritis, gouty arthritis, gingivitis, periodontal disease and other bone metabolic disorders and its It is suitable for suppressing the decrease in bone mass of the reserve army or promoting the increase in bone mass.
  • This product is used as an osteogenesis promoter, osteoporosis, rheumatoid arthritis, rheumatoid spondylitis, Paget's disease, osteoarthritis, gouty arthritis, gingivitis, periodontal disease and other bone metabolic disorders and its It is suitable for suppressing the decrease in bone mass of the reserve army or promoting the increase in bone mass.
  • this product since this product has COX inhibitory activity and nitric oxide synthesis inhibitory activity, it can also be used as a therapeutic or preventive agent for various inflammatory diseases such as rheumatism, cancer, Alzheimer's disease, an analgesic agent and an antipyretic agent. .
  • 2-aminophenol or its derivatives differentiate preosteoblasts into osteoblasts and promote calcium deposition on the cells.
  • 2-aminophenol or a derivative thereof has an action of suppressing osteoclast activity and suppressing bone resorption by an NO synthesis inhibitory action, an IL-6 production inhibitory action, and a PGE 2 synthesis inhibitory action.
  • 2-aminophenol or a derivative thereof is useful as an osteogenesis promoter because it has low toxicity even when taken orally or parenterally and is safe even if used for a long time.

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Abstract

L'invention concerne un promoteur de formation osseuse qui présente une excellente efficacité et peu d'effets secondaires indésirables. Ce promoteur comprend un composé 2-aminophénol ou un dérivé de celui-ci comme principe actif.
PCT/JP2009/057325 2008-04-16 2009-04-10 Promoteur de formation osseuse comprenant un composé 2-aminophénol ou un dérivé de celui-ci comme principe actif Ceased WO2009128395A1 (fr)

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Publication number Priority date Publication date Assignee Title
WO2013047719A1 (fr) * 2011-09-30 2013-04-04 アステラス製薬株式会社 Composé 2-(pyridine-2-yl)pyrimidine-4-amine et sel de celui-ci

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JP2004143101A (ja) * 2002-10-25 2004-05-20 Akio Tomota 抗ウィルス剤
JP2005060325A (ja) * 2003-08-18 2005-03-10 Akio Tomota 抗ヘリコバクター剤、ヘリコバクター属細菌が関与する消化器疾患の予防剤、再発予防剤又は治療剤
JP2005187341A (ja) * 2003-12-24 2005-07-14 Univ Nihon プロスタグランジンｅ２産生抑制剤及び抗炎症剤
JP2005538088A (ja) * 2002-07-22 2005-12-15 オーキッド ケミカルズ アンド ファーマシューティカルズ リミテッド 新規な生物活性分子
JP2007500128A (ja) * 2003-07-25 2007-01-11 アムジエン・インコーポレーテツド 置換複素環式化合物及び使用方法
WO2008047758A1 (fr) * 2006-10-17 2008-04-24 Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo Agent anti-inflammatoire contenant du 2-aminophénol ou un dérivé de celui-ci en tant que principe actif

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JP2005538088A (ja) * 2002-07-22 2005-12-15 オーキッド ケミカルズ アンド ファーマシューティカルズ リミテッド 新規な生物活性分子
JP2004143101A (ja) * 2002-10-25 2004-05-20 Akio Tomota 抗ウィルス剤
JP2007500128A (ja) * 2003-07-25 2007-01-11 アムジエン・インコーポレーテツド 置換複素環式化合物及び使用方法
JP2005060325A (ja) * 2003-08-18 2005-03-10 Akio Tomota 抗ヘリコバクター剤、ヘリコバクター属細菌が関与する消化器疾患の予防剤、再発予防剤又は治療剤
JP2005187341A (ja) * 2003-12-24 2005-07-14 Univ Nihon プロスタグランジンｅ２産生抑制剤及び抗炎症剤
WO2008047758A1 (fr) * 2006-10-17 2008-04-24 Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo Agent anti-inflammatoire contenant du 2-aminophénol ou un dérivé de celui-ci en tant que principe actif

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PHARMACOLOGY, vol. 27, 1983, pages 267 - 280 *

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Publication number Priority date Publication date Assignee Title
WO2013047719A1 (fr) * 2011-09-30 2013-04-04 アステラス製薬株式会社 Composé 2-(pyridine-2-yl)pyrimidine-4-amine et sel de celui-ci

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