Preventive or therapeutic agent for osteoclast proliferative disorders

Abstract

To provide an agent for preventing or treating osteoclast proliferative disease.SOLUTION: An agent for preventing or treating osteoclast proliferative disease contains unfractionated heparin containing polymer heparin as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a preventive or therapeutic agent for osteoclast proliferative disorders. [Background technology]

[0002] Normal bone remodeling is achieved by maintaining a balance between osteoclasts and osteoblasts. Osteoclasts are cells that break down (resorb) bone, while osteoblasts are cells that form bone. If the balance between the two is disrupted and bone resorption by osteoclasts exceeds bone formation by osteoblasts, bone mass decreases and bones become brittle and more susceptible to fracture.

[0003] For such diseases in which the activity of osteoclasts is dominant, drug therapy is generally performed using anti-RANKL antibodies such as denosumab, bisphosphonates, etc. However, although anti-RANKL antibodies and bisphosphonates increase bone mass by suppressing the activity of osteoclasts, there is a problem that side effects such as osteonecrosis of the jaw and femoral fractures occur due to inappropriate bone resorption when administered for a long period of time. In order to solve such problems, new therapeutic drugs are being developed (Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2012-102021 A Summary of the Invention [Problem to be solved by the invention]

[0005] An objective of the present invention is to provide an agent for the prevention or treatment of osteoclast proliferative disorders. [Means for solving the problem]

[0006] The present inventors have conducted intensive research to solve the above problems and have found that heparin has an effect of suppressing differentiation into osteoclasts. Based on these findings, the present inventors have conducted further research and have completed the present invention.

[0007] The present invention encompasses, for example, the subject matter described in the following paragraphs. Section 1. A preventive or therapeutic agent for osteoclast proliferative disorders, comprising unfractionated heparin including high molecular weight heparin as an active ingredient. Section 2. Item 2. The preventive or therapeutic agent according to Item 1, wherein the unfractionated heparin containing high molecular weight heparin is an alkaline earth metal salt of heparin. Section 3. Item 3. The preventive or therapeutic agent according to Item 2, wherein the unfractionated heparin containing high molecular weight heparin is heparin calcium. Section 4. Item 4. The preventive or therapeutic agent according to any one of Items 1 to 3, wherein the osteoclast proliferative disorder is at least one selected from the group consisting of benign or malignant primary bone and soft tissue tumors including giant cell tumor of bone, bone metastasis of cancer, osteoporosis, bone erosion associated with rheumatoid arthritis, Paget's disease of bone, SAPHO syndrome, and chronic recurrent multiple osteomyelitis. Section 5. An osteoclast differentiation inhibitor comprising unfractionated heparin including high molecular weight heparin. Effect of the Invention

[0008] An agent for the prevention or treatment of osteoclast proliferative disorders is provided. [Brief description of the drawings]

[0009] [Figure 1A] Photographs of TRAP staining when heparin calcium was added at various concentrations are shown. [Figure 1B] Photographs of TRAP staining when heparin calcium was added at various concentrations are shown. [Diagram 2] The number of osteoclasts when heparin calcium was added at various concentrations is shown. [Figure 3A]Photographs of TRAP staining in mice administered 0 IU / kg of heparin calcium are shown. [Figure 3B] Photographs of TRAP staining in mice administered 100 IU / kg of heparin calcium are shown. [Figure 3C] Photographs of TRAP staining in mice administered 500 IU / kg of heparin calcium are shown. [Figure 4] Photographs of the tumor in a patient with local recurrence and pulmonary metastasis of giant cell tumor of bone are shown before heparin calcium administration (upper left), after heparin calcium administration (upper right), after switching to warfarin (lower left), and after re-administration of heparin calcium (lower right). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Each embodiment included in the present invention will be described in further detail below.

[0011] The present invention includes a preventive or therapeutic agent for osteoclast proliferative disorders, which contains unfractionated heparin including high molecular weight heparin as an active ingredient. In this specification, the preventive or therapeutic agent for osteoclast proliferative disorders may be referred to as the "preventive or therapeutic agent of the present invention."

[0012] Heparin is an acidic mucopolysaccharide having a repeating structure of uronic acid and glucosamine. Examples of heparin include, but are not limited to, those purified from natural products, commercially available products, etc. Examples of heparin include those isolated from the liver, lung, intestinal mucosa, etc. of healthy pigs, cows, etc.

[0013] In this specification, "unfractionated heparin" refers to heparin having a wide molecular weight distribution, for example, about 6000 to 20000. The upper or lower limit of the molecular weight may be, for example, about 7000, 8000, 9000, 10000, 12000, 14000, 16000, or 18000. More specifically, it may be about 7000 to 18000. Examples of unfractionated heparin include heparin that has not been degraded by enzymatic or chemical treatment.

[0014] The unfractionated heparin used in the present invention may have a number average molecular weight of, for example, about 6000 to 20000. The upper or lower limit of the number average molecular weight may be, for example, about 7000, 8000, 9000, 10000, 12000, 14000, 16000, or 18000. More specifically, it may be about 7000 to 18000.

[0015] As used herein, "high molecular weight heparin" refers to heparin having a molecular weight of, for example, about 6000 or more. The upper or lower limit of the molecular weight may be, for example, about 7000, 8000, 9000, 10000, 12000, 14000, 16000, 18000, or 20000.

[0016] The unfractionated heparin containing high molecular weight heparin used in the present invention includes, for example, unfractionated heparin containing high molecular weight heparin having a molecular weight of about 6000 or more and having a molecular weight of, for example, about 6000 to 20000.

[0017] As used herein, "heparin" also encompasses salts of heparin. The unfractionated heparin including the high molecular weight heparin used in the present invention may be in the form of a salt, for example. Specific examples include alkali metal salts and alkaline earth metal salts. Examples of the alkali metal salts include sodium heparin, and examples of the alkaline earth metal salts include calcium heparin. Among these, alkaline earth metal salts of heparin are preferred, and calcium heparin is particularly preferred. In addition, heparin can be used alone or in combination of two or more types.

[0018] The content of unfractionated heparin including high molecular weight heparin in the preventive or therapeutic agent of the present invention is not particularly limited and can be set appropriately up to a maximum of 100% by mass.

[0019] The preventive or therapeutic agent of the present invention contains the above-mentioned active ingredient and may further contain other ingredients, such as pharma- ceutically acceptable bases, carriers, and / or additives (e.g., solvents, dispersants, emulsifiers, buffers, stabilizers, excipients, binders, disintegrants, lubricants, antioxidants, preservatives, coating agents, colorants, and other drugs such as gastric mucosa protective agents).

[0020] The form of the preventive or therapeutic agent of the present invention is not particularly limited, and examples thereof include tablets, pills, capsules, powders, fine granules, granules, liquids, troches, jellies, injections, plasters, extracts, suppositories, suspensions, tinctures, ointments, poultices, nasal drops, inhalants, liniments, lotions, aerosols, etc. Among these, injections are preferred.

[0021] The preventive or therapeutic agent of the present invention can be prepared by a conventional method by combining the above-mentioned active ingredient with other ingredients as necessary.

[0022] Examples of osteoclast proliferative diseases include benign and malignant primary bone and soft tissue tumors including giant cell tumor of bone, bone metastasis of cancer, osteoporosis, bone erosion associated with rheumatoid arthritis, Paget's disease of bone, SAPHO syndrome, and chronic recurrent multiple osteomyelitis.

[0023] Since the preventive or therapeutic agent of the present invention can suppress differentiation into osteoclasts, it can be used for the prevention or treatment of osteoclast proliferative diseases such as benign or malignant primary bone and soft tissue tumors including giant cell tumor of bone, bone metastasis of cancer, osteoporosis, bone erosion associated with rheumatoid arthritis, Paget's disease of bone, SAPHO syndrome, chronic recurrent multiple osteomyelitis, etc. The osteoclast proliferative diseases may be one type alone or two or more types in combination.

[0024] The subject to which the preventive or therapeutic agent of the present invention is administered is preferably a mammal. It may be not only a human but also a non-human mammal. Examples of the subject include patients with benign or malignant primary bone and soft tissue tumors including giant cell tumor of bone, and humans suspected of benign or malignant primary bone and soft tissue tumors including giant cell tumor of bone; humans with bone metastasis of cancer, and humans suspected of bone metastasis of cancer; cancer patients; patients with primary osteoporosis, patients with secondary osteoporosis caused by drugs, lifestyle-related diseases, rheumatism, nephropathy, and the like, and humans suspected of osteoporosis; patients with rheumatoid arthritis, humans with bone erosion associated with rheumatoid arthritis, and humans suspected of rheumatoid arthritis; patients with Paget's disease of bone, and humans suspected of Paget's disease of bone; patients with SAPHO syndrome, and humans suspected of SAPHO syndrome; patients with chronic recurrent multiple osteomyelitis, and humans suspected of chronic recurrent multiple osteomyelitis. Examples of non-human mammals include mammals kept as pets, livestock, laboratory animals, etc. Examples of such non-human mammals include dogs, cats, monkeys, cows, horses, sheep, goats, pigs, rabbits, mice, rats, camels, llamas, etc.

[0025] Examples of the administration method include oral administration, parenteral (e.g., intravenous, intraarterial, intramuscular, subcutaneous, peritoneal, rectal, transdermal, topical, etc.) administration, etc. Among them, parenteral administration is preferred, and subcutaneous administration and intravenous injection are more preferred.

[0026] The dosage (ingestion) of the preventive or therapeutic agent of the present invention is not particularly limited and is determined depending on the age, sex, severity of symptoms, administration method, etc. of the subject to be administered. For example, the dosage of the active ingredient may be about 50 to 1000 IU / kg body weight per day.

[0027] The preventive or therapeutic agent of the present invention may be administered once a day, or may be administered in divided doses two to three times a day.

[0028] The present invention also includes an osteoclast differentiation inhibitor containing unfractionated heparin including high molecular weight heparin. The osteoclast differentiation inhibitory effect means the effect of inhibiting the differentiation of precursor cells into osteoclasts. The differentiation of precursor cells into osteoclasts can be confirmed by TRAP staining of osteoclasts as shown in the examples described below.

[0029] In addition, in this specification, the term "comprising" includes "consisting essentially of" and "consisting of." In addition, the present invention includes all arbitrary combinations of the constituent elements described in this specification.

[0030] Furthermore, the various characteristics (properties, structures, functions, etc.) described in each embodiment of the present invention above may be combined in any way to specify the subject matter included in the present invention. In other words, the present invention includes all subject matter consisting of all combinations of the combinable characteristics described in this specification. EXAMPLES

[0031] The present invention will be specifically described using the following experimental examples. However, the present invention is not limited to these. In the following, unless otherwise specified, the experiments are performed under atmospheric pressure and room temperature conditions. Furthermore, unless otherwise specified, "%" means "volume %".

[0032] (1) In vitro inhibition of osteoclast differentiation Mouse osteoclast precursor-like cell line RAW264.7 was cultured in a 96-well plate at 5x10 3The cells were seeded at a concentration of 10 ...

[0033] It was confirmed that the number of TRAP-positive cells (osteoclasts) decreased in proportion to the heparin concentration, indicating that the addition of heparin inhibited differentiation into osteoclasts.

[0034] (2) In vivo experiment to inhibit osteoclast expression in mice 12-week-old C57BL / 6 mice were fed a vitamin D-deficient diet for 8 weeks until they reached 20 weeks of age. From 20 weeks of age, they were divided into three groups and administered heparin calcium with molecular weights of 6000-20000 at 0 (5 mice), 100 (4 mice), or 500 (4 mice) IU / kg subcutaneously once a day until they reached 32 weeks of age, at which point their femurs were removed. The vitamin D-deficient diet was continued until they reached 32 weeks of age. After formalin fixation, TRAP staining was performed to evaluate osteoclasts expressed in the distal femoral metaphysis to the epiphysis. The results are shown in Figure 3.

[0035] It was confirmed that the number of TRAP-positive cells (osteoclasts) decreased in proportion to the heparin concentration, indicating that the addition of heparin inhibited differentiation into osteoclasts.

[0036] (3) The therapeutic effect of heparin calcium administration in patients with local recurrence and pulmonary metastasis of giant cell tumor of bone In a patient with local recurrence and lung metastasis of giant cell tumor of bone, the lesions were significantly reduced when 40,000 IU of heparin calcium with a molecular weight of 6,000 to 20,000 was administered subcutaneously once a day. However, when heparin calcium was changed to oral warfarin, the lesions increased. After that, when the administration was returned to 40,000 IU of heparin calcium with a molecular weight of 6,000 to 20,000 subcutaneously once a day, the tumors were reduced again. The dose of heparin calcium was reduced to 20,000 IU subcutaneous injection twice a week, but the tumors remained reduced. Photographs of the tumor before heparin calcium administration (upper left), after heparin calcium administration (upper right), after changing to warfarin (lower left), and after heparin calcium administration again (lower right) are shown in Figure 4.

Claims

1. A therapeutic agent for giant cell tumor of bone, containing unfractionated heparin including high molecular weight heparin as an active ingredient (excluding therapeutic agents containing nanoparticles containing heparin and berberine).

2. The method of claim 1, wherein the unfractionated heparin containing high molecular weight heparin is an alkaline earth metal salt of heparin.

3. The therapeutic agent according to claim 2 , wherein the unfractionated heparin containing high molecular weight heparin is heparin calcium.

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