CN116528900A

CN116528900A - Drug efficacy enhancer for therapeutic agent for inflammatory disease and aggregation promoter for promoting aggregation of contrast agent to inflammatory site

Info

Publication number: CN116528900A
Application number: CN202180067412.5A
Authority: CN
Inventors: 森正树; 山本浩文; 吴鑫
Original assignee: Ultra Micro Yue Co ltd
Current assignee: Ultra Micro Yue Co ltd
Priority date: 2020-09-30
Filing date: 2021-09-30
Publication date: 2023-08-01
Also published as: WO2022071496A1; JPWO2022071496A1

Abstract

The purpose of the present invention is to provide a medicament which can enhance the efficacy of a therapeutic agent for inflammatory diseases. The therapeutic effect of inflammatory diseases can be greatly enhanced by administering a carbonic apatite which does not contain an inflammatory disease therapeutic agent to a patient suffering from inflammatory diseases, in addition to the inflammatory disease therapeutic agent.

Description

Drug efficacy enhancer for therapeutic agent for inflammatory disease and aggregation promoter for promoting aggregation of contrast agent to inflammatory site

Technical Field

The present invention relates to an enhancer for enhancing the efficacy of a therapeutic agent for inflammatory diseases. The present invention also relates to a module for treating inflammatory diseases using the enhancer. The present invention further relates to an aggregation accelerator capable of promoting aggregation of a contrast agent into an inflammatory site and imaging the inflammatory site. The present invention also relates to an imaging module for an inflammatory site using the aggregation promoter.

Background

For inflammatory diseases such as rheumatoid arthritis, various therapeutic agents have been developed with recent advances in medicine. For example, as a therapeutic agent for rheumatoid arthritis, there are put into practical use: etanercept (etanercept) and infliximab

TNF inhibitors such as (infliximab), adalimumab (adalimumab), golimumab (golimumab), cetuzumab (certolizumab pegol), and the like; anti-IL-6 receptor antibodies such as tocilizumab (Tocilizumab) and Sha Lilu monoclonal antibody (sarilumab); and T cell selective co-stimulatory modulators such as abatacept. However, in the conventional therapeutic agents for inflammatory diseases, a certain effectiveness was confirmed, and on the other hand, a sufficient drug effect was not confirmed. Therefore, development of a drug for enhancing the efficacy of a therapeutic agent for inflammatory diseases is demanded.

In addition, in diagnosing inflammatory diseases and in planning treatment, it is important to perform imaging diagnosis of inflammatory sites and accurately determine the position and size of inflammatory sites. Therefore, there is a need to develop a technique capable of efficiently imaging an inflammation site.

On the other hand, it is reported that apatite carbonate has an effect of enhancing an antitumor effect of an anticancer agent, an effect of promoting aggregation of a contrast agent into a tumor, and the like (patent documents 1 and 2). In addition, composite particles in which miR-29a and/or miR-29b are supported on carbonate apatite particles have been reported to be effective in the treatment of inflammatory bowel disease (patent document 3). However, in the treatment of inflammatory diseases, there has been no report on administration of carbonic acid apatite without compounding with a therapeutic agent for inflammatory diseases or use of carbonic acid apatite for imaging an inflammatory site.

Prior art literature

Patent literature

Patent document 1: international publication No. 2015/125934

Patent document 2: japanese patent application laid-open No. 2015-151377

Patent document 3: international publication No. 2018/199121

Disclosure of Invention

Problems to be solved by the invention

The purpose of the present invention is to provide a medicament which can enhance the efficacy of a therapeutic agent for inflammatory diseases. Another object of the present invention is to provide a therapeutic module for treating inflammatory diseases using the agent.

Further, another object of the present invention is to provide an aggregation accelerator capable of imaging an inflammatory site by aggregating a contrast agent at the inflammatory site by a simple method. Another object of the present invention is to provide an imaging module for an inflammatory site using the aggregation promoter.

Means for solving the problems

As a result of intensive studies to solve the above problems, the present inventors have found that the therapeutic effect of inflammatory diseases can be greatly enhanced by administering, in addition to an inflammatory disease therapeutic agent, a carbonic acid apatite which does not contain an inflammatory disease therapeutic agent to a patient suffering from inflammatory diseases. Further, the present inventors have found that by administering not only a contrast agent but also an apatite carbonate containing no contrast agent, aggregation of the contrast agent to an inflammatory site can be promoted, and thus, high-precision imaging of the inflammatory site can be performed. The present invention has been completed based on this finding and has been further studied repeatedly.

That is, the present invention provides the following disclosed embodiments.

The drug efficacy enhancer of the therapeutic agent for inflammatory diseases according to item 1, which comprises as an active ingredient, apatite carbonate not containing the therapeutic agent for inflammatory diseases.

Item 2. The efficacy enhancer of item 1, which is used for enhancing the efficacy of a therapeutic agent for rheumatoid arthritis.

Item 3. The pharmacodynamic enhancer of item 2, wherein the therapeutic agent for rheumatoid arthritis is a TNF inhibitor.

A therapeutic kit for inflammatory diseases comprising a first preparation comprising a therapeutic agent for inflammatory diseases and a second preparation comprising the drug efficacy enhancer according to any one of items 1 to 3,

the first and second formulations are administered separately.

The method of item 5, an aggregation accelerator for an inflammatory site comprising, as an active ingredient, a carbonic apatite which does not contain a contrast agent.

Item 6. The aggregation-promoting agent according to item 5, wherein the contrast agent is a fluorescent contrast agent.

The aggregation-promoting agent according to item 6, wherein the fluorescent contrast agent is indocyanine green.

An inflammatory site imaging device comprising a first preparation containing a contrast agent and a second preparation containing the aggregation accelerator according to any one of items 5 to 7,

The first and second formulations are administered separately.

A method for treating an inflammatory disease, comprising the steps of: a first preparation comprising a therapeutic agent for inflammatory diseases and a second preparation comprising the drug efficacy enhancer according to any one of items 1 to 3 are administered to a patient suffering from inflammatory diseases, respectively.

The use of a carbonic apatite which does not contain a therapeutic agent for inflammatory diseases for producing a drug efficacy enhancer for therapeutic agents for inflammatory diseases.

The method of treating an inflammatory disease, comprising administering to a subject in need thereof an effective amount of a therapeutic agent for inflammatory disease.

A method of imaging an inflammatory site, comprising the steps of: a preparation I comprising a contrast agent and a preparation II comprising the aggregation accelerator according to any one of items 5 to 7 are administered to a subject in need of imaging an inflammatory site, respectively.

Item 13. Use of a contrast agent-free apatite carbonate for the manufacture of an aggregation promoter for a contrast agent to an inflammatory site.

Item 14. A carbonate apatite comprising no contrast agent, for use in a treatment for promoting aggregation of contrast agent to an inflammatory site.

Effects of the invention

According to one embodiment of the present invention, the efficacy of a therapeutic agent for inflammatory diseases can be greatly enhanced. Therefore, according to the present invention, it is expected to improve the therapeutic effect against inflammatory diseases and bring good news to patients suffering from inflammatory diseases.

In addition, according to another aspect of the present invention, accumulation of a contrast medium to an inflammatory site can be promoted by a simple method of administering apatite carbonate, and the inflammatory site can be visualized and imaged. Therefore, according to the present invention, diagnosis of inflammatory diseases, planning of treatment of inflammatory diseases, detection of inflammatory sites in experimental animals, and the like can be performed with high accuracy by a simple method, and thus, the present invention can greatly contribute to technical progress in the fields of diagnosis and treatment of inflammatory diseases.

Drawings

Fig. 1 is a result of observation and scoring of swelling of foot joints in a naturally occurring rheumatoid arthritis model and a non-pathogenic/moderately pathogenic model in test example 1.

Fig. 2 shows the results of the observation of foot joints after 5 hours and 8 hours in test example 1, in which indocyanine green (ICG) was administered intraperitoneally (ICG (i.p.)) or apatite carbonate (sCA) was administered intravenously and ICG was administered intraperitoneally (sCA (i.v.)) +icg (i.p.)) in a naturally occurring rheumatoid arthritis model and a non-pathogenic wind-damp disease model. The upper image is an image of the foot joint, and the lower image is an image obtained by fluorescence imaging of the foot joint.

Fig. 3 shows the results of quantifying fluorescence signals of the foot joints after 5 hours and after 8 hours in test example 1, in which ICG (i.p.) or sCA was administered intraperitoneally and ICG (sCA (i.v.) and ICG (i.p.) was administered intraperitoneally) to a naturally occurring rheumatoid arthritis model.

Fig. 4 a shows the results of RNA analysis of tissues around the bipedal joint after 1 hour and 2 hours by intravenous administration sCA to a model of rheumatoid arthritis with natural onset in test example 2. Further, b in fig. 4 is a result of performing an IPA Upsteam analysis on 845 genomes whose fluctuation was confirmed in a in fig. 4.

Fig. 5 shows the results of fluorescence imaging of inflammatory sites after 30 minutes, after 60 minutes, after 90 minutes, and after 150 minutes in test example 3 for a naturally occurring rheumatoid arthritis model, i.g., ICG (ICG i.p.), or freeze-dried sCA by i.v., and ICG (sCA i.v. + ICG i.p.) by i.c. administration.

Fig. 6 shows the results of quantifying fluorescence signals at the inflammatory sites after 30 minutes, after 60 minutes, after 90 minutes and after 150 minutes in test example 3, for a naturally occurring rheumatoid arthritis model, i.g., ICG (ICG i.p.), or freeze-dried for i.v. administration sCA and i.p. administration ICG (sCA i.v. + ICG i.p.).

Fig. 7 shows the results of quantifying the fluorescent signal of the foot joint over time in the case of administering ICG intraperitoneally (ICG i.p.) or intravenously (sCA) to a model of rheumatoid arthritis with natural onset and administering ICG intraperitoneally after 100 minutes (sCA i.v. (-100 min) +icg i.p.) in test example 4.

Fig. 8 shows the results of quantitative determination of fluorescent signals of foot joints over time in test example 5 by administering ICG (ICG i.p.) to a naturally occurring rheumatoid arthritis model, or by administering ICG (sCA i.v. + ICG i.p.) to the abdominal cavity by freeze-drying sCA.

Fig. 9 a shows the results of quantitative fluorescence signals of the foot joints over time in the case of administering ICG intraperitoneally (ICG i.p.) or in the case of administering ICG intraperitoneally (1/2 scca i.v. (-100 min) +icg i.p.) after 100 minutes of freeze-drying sCA in test example 6. Fig. 9 b shows the results of quantitative fluorescence signals of the foot joints over time in the ICG i.p. group and 1/2 scca i.v. (-100 min) +icg i.p. group in test example 6, from the first administration of ICG to 103.5 hours after the administration of ICG alone.

Fig. 10 a is an image of the hind legs of a mouse with arthritis onset observed in test example 7. Fig. 10 b is a graph showing the results of measuring the width of the feet of the arthritic disease with time in test example 7, in which mice having the arthritic disease were divided into untreated groups (Con), etanercept (Etanercept i.p.), sCA (sCA i.v.), and a combination of sCA and Etanercept (scai.v. + Etanercept i.p.). The arrow in b of fig. 10 refers to the date of administration.

FIG. 11 shows the results of treatment of untreated mice with rheumatoid arthritis (RA (-)), untreated mice with rheumatoid arthritis (RA (+), etanercept-administered mice with rheumatoid arthritis (RA (+) Etanercept i.p.), a combination of sCA and Etanercept (RA (+) sCA) in test example 8

V. + Etanercept i.p.), and sCA administration group (RA (+) sCA i.v.) of rheumatoid arthritis-onset mice, the results of the extent of symptoms of rheumatoid arthritis were observed after the initial administration day (day 0) to 98 days of the drug.

Detailed Description

1. Drug effect enhancer of inflammatory disease therapeutic agent

The drug efficacy enhancer of the present invention is used for enhancing the efficacy of inflammatory diseases, and is characterized by comprising as an active ingredient, apatite carbonate which does not contain a therapeutic agent for inflammatory diseases. The efficacy enhancer of the present invention will be described in detail below.

Carbonate apatite

The carbonate apatite has a structure of mixing hydroxyapatite (Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ ) Part of the hydroxyl groups of (2) is CO ₃ The substituted structure is represented by the general formula Ca _10-m X _m (PO ₄ ) ₆ (CO ₃ ) _1-n Y _n A compound represented by the formula (I). Here, X is an element that can partially replace Ca in the apatite carbonate, and examples thereof include Sr, mn, and rare earth elements. m is a positive number of usually 0 to 1, preferably 0 to 0.1, more preferably 0 to 0.01, still more preferably 0 to 0.001. Y is CO in the carbonate apatite which can be partially substituted ₃ Examples of the group or element(s) include OH, F, and Cl. n is a positive number of usually 0 to 0.1, preferably 0 to 0.01, more preferably 0 to 0.001, still more preferably 0 to 0.0001.

The average particle size of the apatite carbonate used in the present invention is not particularly limited as long as it can be administered into a living body and is transferred into cells, and it is usually more than 50nm (for example, more than 50nm and 3000nm or less), preferably 100 to 3000nm, more preferably 100 to 2000nm, still more preferably 200 to 2000nm or 400 to 3000nm, and particularly preferably 400 to 2000nm.

The average particle diameter of the apatite carbonate is a value obtained by measuring by a dynamic light scattering method (DLS) particle measurement. In the case where large particles (for example, having a particle diameter of 5 μm or more) unsuitable for measurement using DLS are present, they are excluded from the measurement object range. In the present specification, the particle diameter means: the particle size of individual particles that can be identified as individual particles when measured using a scanning probe microscope. Therefore, in the case where a plurality of particles are aggregated, their aggregate will be judged as one particle.

Since the drug effect enhancer of the present invention and the therapeutic agent for inflammatory diseases are administered separately, the drug effect enhancer of the present invention uses apatite carbonate which does not contain the therapeutic agent for inflammatory diseases. Here, "the carbonate apatite containing no therapeutic agent for inflammatory diseases" means a carbonate apatite containing no therapeutic agent for inflammatory diseases in the interior of the particle and having no therapeutic agent for inflammatory diseases attached or bonded to the surface of the particle.

The apatite carbonate can be obtained by a known method. For example, it can be prepared by allowing calcium ions, phosphate ions and bicarbonate ions to coexist in an aqueous solution. The concentration of each ion in the aqueous solution is not particularly limited as long as apatite is formed, and can be appropriately set with reference to the following.

The calcium ion concentration in the aqueous solution is usually 0.1 to 1000mM, preferably 0.5 to 100mM, and more preferably 1 to 10mM.

The phosphate ion concentration in the aqueous solution is usually 0.1 to 1000mM, preferably 0.5 to 100mM, and more preferably 1 to 10mM.

The bicarbonate ion concentration in the aqueous solution is usually 1.0 to 10000mM, preferably 5 to 1000mM, and more preferably 10 to 100mM.

As calcium ions, phosphateThe source of supply of ions and bicarbonate ions is not particularly limited as long as they can be supplied in an aqueous solution, and examples thereof include water-soluble salts of these ions. Specifically, caCl can be used ₂ As the calcium ion source, naH can be used ₂ PO ₄ ·2H ₂ O as phosphate ion source, naHCO can be used ₃ As a source of carbonate ions.

The aqueous solution for preparing the apatite carbonate may contain components other than the ion sources and other substances described above as long as the apatite carbonate is formed. For example, ca or CO in the apatite carbonate may be partially substituted or modified by adding fluoride ion, chloride ion, sr, mn, polyethylene glycol (PEG) or the like to the above composition in an aqueous solution ₃ Etc. However, the amount of the fluorine ion, the chlorine ion, sr, mn, PEG and the like to be added is preferably within a range that does not significantly affect the pH solubility and the particle size range of the composite particles to be formed. The aqueous solution for preparing apatite carbonate may be water as a matrix, but various media for cell culture, buffers, and the like may be used.

In the preparation of the apatite to be used in the present invention, the mixing order of the respective ion sources and other substances in the aqueous solution is not particularly limited, and the aqueous solution may be prepared in any mixing order as long as the target apatite can be obtained. For example, a first solution containing calcium ions and other substances may be prepared, while a second solution containing phosphate ions and bicarbonate ions may be additionally prepared, and the first solution and the second solution may be mixed to prepare an aqueous solution.

The apatite carbonate can be obtained by adjusting the pH of an aqueous solution containing the ions to a range of 6.0 to 9.0 and allowing the solution to stand (incubate) for a predetermined period of time. The pH of the aqueous solution at the time of forming the apatite carbonate may be, for example, 7.0 to 8.5, preferably 7.1 to 8.5, more preferably 7.2 to 8.5, still more preferably 7.3 to 8.5, particularly preferably 7.4 to 8.5, and most preferably 7.5 to 8.0.

The temperature conditions of the aqueous solution at the time of forming the apatite carbonate are not particularly limited as long as the apatite carbonate is formed, and examples thereof are usually 4℃or higher, preferably 25 to 80℃and more preferably 37 to 70 ℃.

The incubation time of the aqueous solution for forming the apatite carbonate is not particularly limited as long as the apatite carbonate can be formed, and examples thereof include usually 1 minute to 24 hours, preferably 5 minutes to 1 hour. Whether particles are formed or not can be confirmed, for example, by observation under a microscope.

The method of controlling the average particle size of the apatite to be within the above range is not particularly limited, and examples thereof include a method of subjecting the apatite formed in the aqueous solution to ultrasonic vibration treatment. Specific examples of the ultrasonic vibration treatment include: a treatment of applying ultrasonic waves by bringing an ultrasonic transducer such as an ultrasonic crusher into direct contact with a sample; an ultrasonic washer having an ultrasonic transducer and a water tank (cleaning tank) is used, and a liquid (for example, water) is added to the water tank, a container (for example, a plastic pipe) for containing the apatite carbonate is floated in the liquid, and an ultrasonic wave treatment is applied to an aqueous solution containing the apatite carbonate through the liquid. By such ultrasonic vibration treatment, the particle size of the apatite carbonate can be easily and efficiently reduced to the above range. Further, after the ultrasonic treatment, the particle size of the apatite can be adjusted to the above range by separating the apatite with a desired particle size by a filter having a specific pore size.

The conditions of the ultrasonic vibration treatment are not particularly limited as long as the particle size can be controlled within a predetermined range. For example, when the ultrasonic cleaning is performed using an ultrasonic cleaner having an ultrasonic vibrator and a water tank (cleaning tank), the following conditions can be exemplified.

Temperature of the water tank: for example, the temperature is 5 to 45 ℃, preferably 10 to 35 ℃, and more preferably 20 to 30 ℃.

High frequency output: for example, 10 to 500W, preferably 20 to 400W, more preferably 30 to 300W, and even more preferably 40 to 100W.

Oscillation frequency: for example, 10 to 60Hz, preferably 20 to 50Hz, more preferably 30 to 40Hz.

The treatment time is as follows: for example, 30 seconds to 30 minutes, preferably 1 to 20 minutes, and more preferably 3 to 10 minutes.

The type of the container containing the apatite carbonate used for the ultrasonic vibration treatment is not limited as long as the particles can be made finer to a predetermined particle size range, and may be appropriately selected depending on the volume of the aqueous solution and the purpose of use. For example, plastic tubing having a capacity of 1 to 1000ml may be used.

The ultrasonic vibration treatment is preferably performed in the presence of a dispersant (i.e., in a state in which a dispersant is added to an aqueous solution containing apatite carbonate). The reason for this is that by performing ultrasonic vibration treatment in an environment where a dispersant and apatite coexist, apatite nanoparticles having a finer particle diameter can be obtained, and the re-agglomeration of particles can be suppressed. The type of the dispersant is not particularly limited as long as the carbonate apatite can be dispersed, and the dispersant is usually added to a drug, and examples thereof include albumin. The dispersant may be used alone or in combination of 1 or more than 2. The concentration of the dispersant in the aqueous solution containing the apatite carbonate is not particularly limited as long as the effect of reducing the size and/or suppressing the re-aggregation can be obtained, and examples thereof include about 0.1 to 500mg/ml, preferably about 1 to 100mg/ml, and more preferably about 1 to 10 mg/ml; or about 0.001 to 10 wt%. In this way, the dispersant added to miniaturize the apatite particles can be administered to the living body together with the apatite particles in a state of being contained in the drug efficacy enhancer of the present invention.

The drug efficacy enhancer of the present invention is used in a state in which the apatite carbonate is dispersed in a dispersion of a solvent such as physiological saline. The drug efficacy enhancer of the present invention may be provided as a dispersion of the carbonic acid apatite, or the carbonic acid apatite may be provided in a dried state such as freeze-drying, and may be dispersed in a solvent such as physiological saline for use.

Usage and dosage

The drug efficacy enhancer of the present invention is useful for enhancing the efficacy of a therapeutic agent for inflammatory diseases. In the present invention, the "efficacy of an inflammatory disease therapeutic agent" refers to the preventive, palliative, ameliorative or therapeutic effects of an inflammatory disease caused by an inflammatory disease therapeutic agent.

Examples of the inflammatory disease to be used as the drug efficacy enhancer of the present invention (i.e., the disease to be treated with the enhancer of the inflammatory disease) include chronic inflammatory diseases, autoimmune diseases, allergic diseases, and the like. Among these inflammatory diseases, inflammatory autoimmune diseases are preferred.

Specific examples of inflammatory diseases include rheumatoid arthritis, inflammatory bowel disease, crohn's disease, ulcerative colitis, multiple sclerosis, autoimmune hepatitis, primary biliary cirrhosis, guillain-Barre syndrome, diffuse toxic goiter, toyota disease, pemphigus, multiple sclerosis, systemic lupus erythematosus, sjogren's syndrome, systemic scleroderma, dermatomyositis, psoriasis, vitiligo, vesicular pemphigoid, sudden diastolic cardiomyopathy, type 1 diabetes, hashimoto's disease, myasthenia gravis, igA nephropathy, membranous nephropathy, pernicious anemia, antiphospholipid antibody syndrome, polymyositis, dermatomyositis, scleroderma, igG 4-related diseases, vasculitis syndrome, mixed connective tissue diseases, inflammatory bowel disease, atopic dermatitis, and the like.

Among these inflammatory diseases, rheumatoid arthritis is preferred as a target of application of the efficacy enhancer of the present invention.

In the drug efficacy enhancer of the present invention, the type of the therapeutic agent for inflammatory diseases to be enhanced in drug efficacy is not particularly limited, and may be appropriately selected depending on the type of inflammatory disease. For example, as a therapeutic agent for rheumatoid arthritis, there may be mentioned: TNF inhibitors such as etanercept, infliximab, adalimumab, golimumab (golimumab), cetuximab (golimumab), cetuzumab (certolizumab pegol), and the like; anti-IL-6 receptor antibodies such as tocilizumab (Tocilizumab) and Sha Lilu monoclonal antibody (sarilumab); and T cell selective co-stimulatory modulators such as abatacept. Among these therapeutic agents for rheumatoid arthritis, the efficacy enhancer of the present invention is particularly preferably used for the purpose of enhancing the efficacy of TNF inhibitors (particularly etanercept).

The pharmacodynamic potentiators of the present invention are administered by systemic administration. Examples of systemic administration include intravascular (intra-arterial or intravenous) administration, subcutaneous administration, intraperitoneal administration, etc., preferably intravascular administration, and more preferably intravenous administration. It should be noted that intravascular administration includes not only intravascular injection but also continuous infusion. The method of administration of the drug efficacy enhancer of the present invention may be the same as or different from the method of administration of the therapeutic agent for inflammatory diseases to be enhanced in drug efficacy.

The method of administration of the therapeutic agent for inflammatory diseases having enhanced efficacy by the efficacy enhancer of the present invention is not particularly limited, and may be appropriately set depending on the type of therapeutic agent for inflammatory diseases or the like, and may be the same as or different from the method of administration of the efficacy enhancer of the present invention. Specific examples of the method of administration of the therapeutic agent for inflammatory diseases include intraperitoneal administration, intravascular (intra-arterial or intravenous) administration, subcutaneous administration, and oral administration. It should be noted that intravascular administration includes not only intravascular injection but also continuous infusion.

The amount of the drug effect enhancer of the present invention to be administered is appropriately determined depending on the type of the inflammatory disease therapeutic agent to be enhanced in drug effect, the sex, age, symptoms of the patient, and the like, and thus cannot be generally said, and is not limited to, for example, about 10mg to 1g/kg (body weight) per 1 time in terms of the amount of apatite carbonate.

The amount of the therapeutic agent for inflammatory diseases having an enhanced drug effect by the agent for enhancing drug effect of the present invention may be a therapeutically effective amount, and may be appropriately determined according to the type of inflammatory disease to be treated, the type of therapeutic agent for inflammatory disease to be used, the sex, age, and symptoms of the patient, and the like, and may be, for example, about 1 to 1500mg or about 10 to 1200mg per 1 adult.

The drug effect enhancer of the present invention is administered separately from the therapeutic agent for inflammatory diseases. In the present invention, "administered separately from the therapeutic agent for inflammatory disease" means that the therapeutic agent for inflammatory disease of the present invention and the therapeutic agent for inflammatory disease are not administered as the same preparation (mixture). In this case, the drug effect enhancer of the present invention and the therapeutic agent for inflammatory disease are administered separately.

The administration time of the efficacy-enhancing agent of the present invention is not particularly limited, as long as it is, for example, about 100 hours or less, preferably about 24 hours or less, more preferably about 6 hours or less, and even more preferably about 2 hours or less, of the administration of the therapeutic agent for inflammatory disease to be enhanced.

2. Therapeutic assembly for inflammatory diseases

The therapeutic device for inflammatory diseases according to the present invention is a therapeutic agent set for use in the treatment of inflammatory diseases, and is characterized by comprising a first preparation containing an inflammatory disease therapeutic agent and a second preparation containing the efficacy-enhancing agent, wherein the first preparation and the second preparation are administered separately.

The therapeutic module for inflammatory diseases of the present invention is provided by grouping the aforementioned drug efficacy enhancer and the therapeutic agent for inflammatory diseases. In the module for treating inflammatory diseases according to the present invention, the type of inflammatory diseases to be treated, the type of the first agent (therapeutic agent for inflammatory diseases) and the second agent (the drug efficacy enhancer), the administration method, the amount of the drug to be administered, and the like are as shown in the column "1. Therapeutic agent for inflammatory diseases". Further, since the drug effect enhancer and the therapeutic agent for inflammatory disease are administered separately as described above, the second preparation does not contain the therapeutic agent for inflammatory disease.

3. Agent for promoting aggregation of contrast agent to inflammatory site

The aggregation accelerator of the present invention is used for accelerating aggregation of a contrast agent to an inflammatory site, and is characterized by comprising, as an active ingredient, an apatite carbonate which does not contain a contrast agent. The aggregation accelerator of the present invention will be described in detail below.

Carbonate apatite

The average particle diameter and the production method of the apatite carbonate used in the present invention are as described in the column "1. Pharmacodynamics enhancer of therapeutic agent for inflammatory diseases".

Since the aggregation accelerator of the present invention is administered separately from the contrast agent, the aggregation accelerator of the present invention uses a carbonic apatite which does not contain the contrast agent. Here, "apatite containing no contrast agent" refers to a carbonate apatite containing no contrast agent inside the particles and having no contrast agent attached or bound to the particle surface.

Usage and dosage

The aggregation-promoting agent of the present invention is used for promoting aggregation of a contrast agent at an inflammatory site generated in an inflammatory disease. In the present invention, the type of the contrast agent that promotes accumulation to the inflammatory site is not particularly limited as long as it is cytotoxic and can contrast the tissue in the living body, regardless of the presence or absence of the accumulation ability of the contrast agent itself to the inflammatory site. Even if the contrast agent itself does not have the ability to accumulate in the inflammatory site, the aggregation accelerator of the present invention can cause the contrast agent to accumulate in the inflammatory site and image the inflammatory site.

In the aggregation accelerator of the present invention, specific examples of the contrast agent that aggregates at an inflammatory site include a fluorescent contrast agent, an X-ray contrast agent, an MRI contrast agent, a PET contrast agent, and a SPECT contrast agent. Among these contrast agents, the fluorescent contrast agent has a high accumulation promoting effect on an inflammatory site and is less burdened on a subject, and is therefore suitable as an application target of the accumulation promoting agent of the present invention.

Specific examples of the fluorescent contrast agent include indocyanine green (ICG), fluorescein, quantum dots, near infrared fluorescent dyes (Cy 5.5, cy7, alexaFluoro, etc.), and other fluorescent substances. Indocyanine green is a fluorescent contrast agent that receives excitation light having a wavelength of around 805nm and emits fluorescence having a wavelength of around 835 nm. 5-aminolevulinic acid is a fluorescent contrast agent that emits fluorescence having a wavelength of about 620nm upon receiving excitation light having a wavelength of about 380 nm. Fluorescein is a fluorescent contrast agent that emits fluorescence having a wavelength of 521nm when receiving excitation light having a wavelength of 494 nm. Among these fluorescent contrast agents, indocyanine green is preferable from the viewpoint of more efficiently promoting accumulation to tumors.

Further, as a preferable example of an inflammatory disease to be applied as the aggregation accelerator of the present invention, an inflammatory autoimmune disease can be given. Examples of inflammatory autoimmune diseases include rheumatoid arthritis, inflammatory bowel disease, crohn's disease, ulcerative colitis, multiple sclerosis, autoimmune hepatitis, primary biliary cirrhosis, guillain-Barre syndrome, diffuse toxic goiter, toyota disease, pemphigus, and the like. Among these inflammatory diseases, rheumatoid arthritis is preferred as a target for application of the aggregation-promoting agent of the present invention.

The aggregation promoters of the present invention are administered by systemic administration. Examples of systemic administration include intravascular (intra-arterial or intravenous) administration, subcutaneous administration, intraperitoneal administration, etc., preferably intravascular administration, and more preferably intravenous administration. It should be noted that intravascular administration includes not only intravascular injection but also continuous infusion. The administration method of the aggregation accelerator of the present invention may be the same as or different from that of the contrast agent.

The method of administration of the contrast agent to be accumulated in the inflammatory site by the aggregation accelerator of the present invention is not particularly limited, and may be appropriately set depending on the type of the contrast agent, the inflammatory site, and the like, and may be the same as or different from the method of administration of the aggregation accelerator of the present invention. Even if the administration method of the contrast agent is different from that of the aggregation accelerator of the present invention, accumulation of the contrast agent to the inflammatory site can be promoted by the aggregation accelerator of the present invention. Specific examples of the method of administering the contrast agent include intraperitoneal administration, intravascular (intra-arterial or intravenous) administration, subcutaneous administration, and the like, and intraperitoneal administration is preferable. It should be noted that intravascular administration includes not only intravascular injection but also continuous infusion.

The amount of the aggregation accelerator of the present invention to be administered is appropriately determined depending on the type of the contrast medium to be the target of the acceleration of the accumulation to the inflammatory site, the sex, age, symptoms of the patient, and the like, and thus cannot be approximated, for example, it is only required to convert the amount of apatite carbonate to about 10mg to 1g/kg (body weight) every 1 time.

The amount of the contrast medium to be administered, which is accumulated in the inflammatory site by the aggregation accelerator of the present invention, may be appropriately determined depending on the inflammatory site, the sex, age, symptoms, etc. of the patient, and may be, for example, about 5 to 30mg per 1 adult.

The aggregation promoters of the present invention are administered separately from the contrast agent. In the present invention, "administered separately from a contrast agent" means that the aggregation accelerator of the present invention and the contrast agent are administered without being formulated into the same formulation (mixture). The administration time of the aggregation accelerator of the present invention and the contrast agent may be simultaneous, but in this case, the aggregation accelerator of the present invention and the contrast agent are administered separately.

The administration time of the aggregation accelerator of the present invention is not particularly limited, as long as it is, for example, 100 hours or less before and after administration of the contrast agent, preferably 24 hours or less before and after administration of the contrast agent, more preferably 6 hours or less before and after administration of the contrast agent, and still more preferably 2 hours or less before and after administration of the contrast agent.

By separately administering the aggregation accelerator and the contrast agent of the present invention, the contrast agent is accumulated at the site of inflammation. Thereafter, the inflammatory site can be imaged by a known method according to the type of the contrast agent used. For example, if a fluorescent contrast medium is used, since fluorescence is emitted from the fluorescent contrast medium by irradiation of excitation light, an inflammatory site can be imaged by observing the fluorescence.

The imaging of the inflammatory site using the aggregation accelerator of the present invention is suitably used for diagnosis of inflammatory diseases, confirmation of the degree of progression, confirmation of the inflammatory site in experimental animals, and the like.

4. Imaging module for inflammatory site

The present invention provides a kit for imaging an inflammatory site, which is a kit for use in imaging an inflammatory site, comprising an I-th preparation containing a contrast agent and a II-th preparation containing the aggregation-promoting agent, wherein the I-th preparation and the II-th preparation are administered separately.

The component for imaging an inflammatory site of the present invention is provided by grouping the aggregation accelerator and the contrast agent. In the component for imaging an inflammatory site of the present invention, the type of inflammatory disease to be imaged, the type of the I-th preparation (contrast agent) and the II-th preparation (aggregation accelerator) and the administration method and the administration amount are shown in the column "3. Aggregation accelerator of contrast agent to inflammatory site". In addition, since the aggregation accelerator is administered separately from the contrast agent as described above, the second preparation does not contain the contrast agent.

Examples

The present invention will be described below with reference to examples. However, the present invention is not limited to the following examples.

Reference example 1: production of apatite carbonate (sonicated carbonate apatite; sCA)

First, an inorganic aqueous solution (NaHCO) ₃ ；44mM，NaH ₂ PO ₄ ；0.9mM，CaCl ₂ The method comprises the steps of carrying out a first treatment on the surface of the 1.8mM, pH 7.5), incubated at 37℃for 30 minutes. Then, precipitation was performed at 12000rpm×3 minutes, whereby particles of apatite carbonate (sCA) were obtained. To the obtained pellets, physiological saline (containing 0.5 wt% albumin) was added to 200. Mu.L and dispersed, and the pellets were subjected to ultrasonic vibration treatment (38 kHz, 80W) for 10 minutes, immediately used for the test, or lyophilized. The average particle diameter of sCA obtained was found to be 400 to 2000nm in dynamic light scattering particle measurement (DLS) using a Zetasizer NanoZS (Malvern).

Test example 1: imaging of fluorescent substances at inflammatory sites using a model of rheumatism

Mannans (purchased from sigma) were intraperitoneally administered (20 mg/500. Mu.l physiological saline/mouse) to 8-week-old SKG/Jcl mice (females, purchased from Japanese cleara) and, after several weeks, autoimmune arthritis very similar to human rheumatoid arthritis (Rheumatoid Arthritis: RA) was developed and a model of naturally-occurring rheumatoid arthritis was established. The degree of swelling of the foot joints of naturally occurring rheumatoid arthritis model and mice not administered with mannan (rheumatism non-occurring model) was scored according to the following criteria. In the following test examples, the inflammation scores were also based on the following criteria. The total of the inflammation scores is a total value of the inflammation scores of the respective swelling sites confirmed from the mice.

Baseline of score of extent of swelling of foot joints (inflammation score, RA score)

0: no swelling

0.5: moderate swelling

1: severe swelling

Swelling of the foot joints in the naturally occurring rheumatoid arthritis model and in the non-occurring rheumatic disease model was observed, and the scoring results are shown in fig. 1.

For the naturally occurring rheumatoid arthritis model and the non-occurring rheumatic disease model established above, 200 μl of physiological saline (i.p.) containing 1.25mg/ml indocyanine green (ICG) was administered intraperitoneally (i.p.), and after 5 hours and after 8 hours, imaging (ICG (i.p.) groups) of foot joints were performed by fluorescence imaging using IVIS Spectrum. Further, after 3 days thereof, physiological saline containing sCA (containing sCA in an amount corresponding to 50ml of the inorganic aqueous solution prepared in reference example 1 described above) was intravenously administered (i.v.) and 200 μl of physiological saline containing 1.25mg/ml of ICG was intraperitoneally administered (i.p.) and after 5 hours and after 8 hours, imaging of the foot joint was performed by fluorescence imaging using IVIS Spectrum (sCa (i.v.) group+icg (i.p.).

FIG. 2 shows the results of ICG detection by fluorescence imaging using IVIS Spectrum. As a result, in the naturally occurring rheumatoid arthritis model with an inflammation score of 2, the fluorescence signal intensity at the foot joints of the sCA (i.v.) group+icg (i.p.) group was stronger than that of the ICG (i.p.) group. Fig. 3 shows the result of quantifying the fluorescence signal of the foot joint in the naturally occurring rheumatoid arthritis model. As a result, it was confirmed that the fluorescence intensity was significantly higher in the sCA (i.v.) +icg (i.p.) group than in the ICG (i.p.) group.

Test example 2: mechanism of aggregation-enhancing effect of agent at inflammation site based on sCAA naturally occurring rheumatoid arthritis model (total inflammation score of 2) was prepared in the same manner as in test example 1. In a naturally occurring rheumatoid arthritis model, normal saline (containing sCA in an amount equivalent to 50ml of an inorganic aqueous solution prepared in reference example 1 described above) containing sCA was administered intravenously (i.v.), and tissues surrounding the bipedal joint of the same mouse were collected by killing them 1 hour and 2 hours after administration. In addition, the same amount of physiological saline was intravenously administered without sCA, and after 1 hour and 2 hours from the administration, the tissues around the bipedal joint of the same mouse were sacrificed and collected (untreated group).

RNA analysis of collected bipedal joint peripheral tissues was performed on each of untreated groups (mice n=4), sCA groups 1 hour after intravenous administration (mice n=3), and sCA groups 2 hours after intravenous administration (mice n=3). The results are shown in fig. 4 a. As a result, it was found that 845 genes with a Fold change (Fold change) of ±2 and p < 0.05 were observed in the group after the administration of sCA vein, compared with the untreated group, and these genes were changed with time. Further, the results of upstream (Upsteam) analysis of the 845 genomes by specific pathway analysis (Ingenuity Pathways Analysis (IPA)) are shown in fig. 4 b. As a result, it was predicted that H was administered intravenously through sCA ₂ O ₂ Upstream regulatory elements (Upstream regulator) such as, NFkB, EGF, VEGF, prostaglandin (prostaglandin) E2, active oxygen (reactive oxygen species), TNF, PTGS2, NOS1, etc. These factors are known as factors that increase vascular permeability.

Test example 3: fluorescent substance aggregation enhancement effect to inflammation site based on freeze-drying sCA-through sCA at the same time Time-lapse imaging of drug administration

A naturally occurring rheumatoid arthritis model (total inflammation score of 2) was prepared in the same manner as in test example 1. The model of rheumatoid arthritis with natural onset used in this test also had a plurality of joints at the tail, and was evaluated as an inflammatory site.

For the above-mentioned naturally occurring rheumatoid arthritis model, 500 μl of physiological saline containing 0.83mg/ml ICG was administered intraperitoneally (i.p.), and imaging of the foot joints was performed by fluorescence imaging using IVIS Spectrum (ICG i.p. group) 30 minutes after, 60 minutes after, 90 minutes after, and 150 minutes after the administration. In addition, for the above-mentioned model of naturally occurring rheumatoid arthritis, imaging of the inflammatory site was performed by fluorescence imaging using IVIS Spectrum (sCA i.v. + ICG i.p. group) by administering 500 μl of physiological saline containing 0.83mg/ml ICG to the abdominal cavity (i.p.) while administering (i.v.) physiological saline containing freeze-dried sCA (physiological saline containing sCA in an amount corresponding to 30ml prepared from the inorganic aqueous solution in reference example 1) intravenously (i.v.) for 30 minutes, 60 minutes, 90 minutes, and 150 minutes from the administration).

FIG. 5 shows the results of ICG detection by fluorescence imaging using IVIS Spectrum. As a result, it was found that the sCA i.v. + ICG i.p. group had a stronger fluorescence signal intensity at the inflammatory site than the ICG i.p. group in the naturally occurring rheumatoid arthritis model. Fig. 6 shows the result of quantifying the fluorescent signal at the inflammatory site. As a result, it was confirmed that in the sCA i.v. + ICG i.p. group, the fluorescence signal intensity in the inflammatory site was significantly higher than in the ICG i.p. group.

Test example 4: aggregation enhancement effect of fluorescent substance on inflammation site based on freeze-dried sCA-by Time-lapse imaging of sCA min pre-administration

A naturally occurring rheumatoid arthritis model (total inflammation score of 2) was prepared in the same manner as in test example 1.

For the above-mentioned naturally-occurring rheumatoid arthritis model, 500 μl of physiological saline containing 0.83mg/ml ICG was administered intraperitoneally (i.p.), and after 70 hours from the administration, imaging of the foot joint was performed by fluorescence imaging using IVIS Spectrum, and the fluorescence signal intensity at the foot joint was quantified (ICG i.p. group). In addition, for the above-mentioned model of naturally occurring rheumatoid arthritis, intravenous administration (i.v.) contained freeze-dried sCA physiological saline (containing sCA physiological saline in an amount equivalent to 30ml prepared from the inorganic aqueous solution in reference example 1 above), 100 minutes after the administration of sCa, intraperitoneal administration (i.p.) 500. Mu.l of physiological saline containing 0.83mg/ml of ICG, imaging of the foot joint by fluorescence imaging using IVIS Spectrum was performed over time from the start of ICG administration to 70 hours, and the fluorescence signal intensity at the foot joint was quantified (sCA i.v. (-100 min) +ICG i.p. group).

The results are shown in FIG. 7. As a result, the relative fluorescence intensity of the foot joints in sCA i.v. (100 min) +icg i.p. group was higher than that in ICG i.p. group. That is, it was found that ICG can be efficiently accumulated in the inflammatory site even when sCa is administered 100 minutes before ICG is administered.

Test example 5: fluorescent substance aggregation enhancement effect to inflammation site based on freeze-drying sCA-through sCA at the same time Time-lapse imaging of drug administration

For the above-mentioned naturally-occurring rheumatoid arthritis model, 500 μl of physiological saline containing 0.83mg/ml ICG was administered intraperitoneally (i.p.), and 45 hours after administration, imaging of the foot joint was performed by fluorescence imaging using IVIS Spectrum, and the fluorescence signal intensity at the foot joint was quantified (ICG i.p. group). In addition, for the above-mentioned model of rheumatoid arthritis with natural onset, while normal saline (containing sCA in an amount equivalent to 30ml of an inorganic aqueous solution prepared in reference example 1) containing lyophilized sCA was administered intravenously (i.v.), 500 μl of normal saline containing 0.83mg/ml ICG was administered intraperitoneally (i.p.) and imaging of the foot joint was performed by fluorescence imaging using IVIS Spectrum over time after 45 hours from the administration, and the fluorescence signal intensity at the foot joint was quantified (sCA i.v. + ICG i.p. group).

The results are shown in FIG. 8. As a result, the relative fluorescence intensity of the foot joints in the sCA i.v. + ICG i.p. group was higher than that in the ICG i.p. group.

Test example 6: fluorescent substance aggregation enhancement effect-sCA decrease and increase to inflammation site based on freeze-drying sCA Evaluation of Strong action persistence

For the above-described naturally-occurring rheumatoid arthritis model, 500 μl of physiological saline containing 0.83mg/ml ICG was administered intraperitoneally (i.p.) and imaging of the foot joints was performed by fluorescence imaging using IVIS Spectrum over time, and the fluorescence signal intensity at the foot joints (first time) was quantified (ICG i.p. group). Further, in the ICG i.p. group, 500 μl of physiological saline containing 0.83mg/ml ICG was administered intraperitoneally (i.p.) again after 103.5 hours from the start of the first ICG administration, and similarly, the fluorescence signal intensity at the foot joint (second time) was quantified over time.

In addition, for the above-mentioned model of naturally occurring rheumatoid arthritis, a physiological saline solution (i.v.) containing freeze-dried sCA (containing sCA equivalent to 15ml of an inorganic aqueous solution prepared in the above-mentioned reference example 1; half the amount as compared to sCa of the above-mentioned test example 4) was intravenously administered, and after 100 minutes from sCa administration, 500. Mu.l of a physiological saline solution (i.p.) containing 0.83mg/ml of ICG was intraperitoneally administered (i.p.), and imaging of the foot joint was performed by fluorescence imaging using IVIS Spectrum, and the fluorescence signal intensity at the foot joint was quantified (1/2 sCA i.v. (-100 min) +ICG i.p. group). Further, in the 1/2sCA i.v. (-100 min) +ICG i.p. group, after 103.5 hours from the first ICG administration, 500. Mu.l of physiological saline containing 0.83mg/ml of ICG was administered again intraperitoneally (i.p.) and similarly, the fluorescence signal intensity at the foot joint (second time) was quantified over time.

The results of measuring the fluorescence signal intensity at the foot joint with time after the first ICG administration are shown in fig. 9 a, and the results of measuring the fluorescence signal intensity at the foot joint with time after the second ICG administration are shown in fig. 9 b. From this result, it was confirmed that even when the dose of sCA was reduced, the effect of enhancing the aggregation of ICG produced by sCA to the inflammatory site was confirmed, and further that the effect of enhancing the aggregation of ICG produced by sCA to the inflammatory site was continued for about 100 hours from the administration of sCA.

Test example 7: research on sCA efficacy enhancement effect of antibody-induced collagen arthritis mouse model

Arthritis-initiating monoclonal antibody cocktails (Arthrogen-CIA arthritis-initiating monoclonal antibodies Chondrex Corp.) were intravenously administered as sensitizers to 8-week-old female DBA-1 mice (Charles River Laboratories Japan, inc.) at a dose of 75mg/kg, and LPS (lipopolysaccharide) was intraperitoneally administered at a dose of 1.25mg/kg after 6 days. After 7 days from LPS administration, as in FIG. 10 a, arthritis was developed in the right foot of the mice, and swelling was observed over a wide range.

Mice with arthritis onset (mice 7 days after LPS administration, day 0) were divided into 4 groups shown in Table 1, and drug administration was performed under each condition, and the width of feet with arthritis onset was measured over time from the drug administration (day 0) to 6 days, and the therapeutic effect of each group was evaluated.

TABLE 1

The results are shown in fig. 10 b. As a result, the tendency of swollen feet to shrink is most pronounced in the sCA and etanercept combined group, with a significantly smaller width at day 6 than in the other groups. (One Way ANOVA with Tukey's multiple comparisons test: convs. sCA i.v. + EtOnercept i.p.p <0.001;Etanercept i.p.vs.sCA i.v.+ EtOnercept i.p.p <0.05, sCA i.v. + EtOnercept i.p.vs. sCA i.v. p < 0.05).

Test example 8: research on enhancement effect of therapeutic effect of sCA on model of naturally occurring rheumatoid arthritis

To SKG/Jcl mice (female, purchased from Japanese cleara) of about 8 weeks old, mannans (purchased from sigma) were intraperitoneally administered (20 mg/500. Mu.l physiological saline/mouse), and rheumatoid arthritis-onset mice having a total inflammatory score of 2, which were very similar to human rheumatoid arthritis (Rheumatoid Arthritis: RA) in immunopathology, were prepared 19 days after administration of mannans (day 0). In addition, mice in which rheumatoid arthritis was not developed without administering mannan to SKG/Jcl mice of about 8 weeks of age and without developing autoimmune arthritis were prepared separately.

The mice with rheumatoid arthritis and the mice without rheumatoid arthritis were divided into 5 groups shown in table 2, and the drug was administered under each condition, and the degree of symptoms of rheumatoid arthritis was observed in each mouse from the initial administration day (day 0) to 98 days later.

TABLE 2

The results are shown in FIG. 12. As a result, in the RA (+) group, swelling, stiffness, deformation, and contracture of the limb finger were observed in the joint, as compared with the RA (-) group. In 3 mice suffering from rheumatoid arthritis in the RA (+) Etanercept i.p. group, the symptoms of rheumatoid arthritis were not improved. On the other hand, in the RA (+) sCA i.v. + Etanercept i.p. group, the symptoms of rheumatoid arthritis were significantly improved. That is, according to the present results, sCa has an effect of enhancing the therapeutic effect of etanercept. Although etanercept is usually administered 2 times per week and 3.0mg/kg, the combination with sCa confirms a sufficient therapeutic effect even if etanercept is administered 1 time per month.

Claims

1. A drug effect enhancer for an inflammatory disease therapeutic agent, characterized by comprising as an active ingredient an apatite carbonate which does not contain an inflammatory disease therapeutic agent.

2. The efficacy enhancer of claim 1, wherein the efficacy enhancer is for enhancing the efficacy of a therapeutic agent for rheumatoid arthritis.

3. The potentiator of claim 2, wherein the therapeutic agent for rheumatoid arthritis is a TNF inhibitor.

4. A therapeutic kit for inflammatory diseases, comprising a first preparation comprising a therapeutic agent for inflammatory diseases and a second preparation comprising the drug efficacy enhancer according to any one of claims 1 to 3, wherein the first preparation and the second preparation are administered separately.

5. An aggregation-promoting agent for an inflammatory site, characterized by comprising, as an active ingredient, an apatite carbonate which does not contain a contrast agent.

6. The aggregation-promoting agent according to claim 5, wherein the contrast agent is a fluorescent contrast agent.

7. The aggregation-promoting agent according to claim 6, wherein the fluorescent contrast agent is indocyanine green.

8. An assembly for imaging an inflammatory site, comprising a first preparation containing a contrast agent and a second preparation containing the aggregation accelerator according to any one of claims 5 to 7, wherein the first preparation and the second preparation are administered separately.

9. A method for treating an inflammatory disease, comprising the steps of: a first preparation comprising a therapeutic agent for inflammatory diseases and a second preparation comprising the drug efficacy enhancer according to any one of claims 1 to 3 are administered to a patient suffering from inflammatory diseases, respectively.

10. Use of carbonic apatite not containing an inflammatory disease therapeutic agent for producing a drug effect enhancer for an inflammatory disease therapeutic agent.

11. A carbonic apatite which does not contain an inflammatory disease therapeutic agent, characterized by being used for a treatment for enhancing the efficacy of an inflammatory disease therapeutic agent.

12. A method for imaging an inflammatory site, comprising the steps of: a preparation I comprising a contrast agent and a preparation II comprising the aggregation accelerator according to any one of claims 5 to 7 are administered to a subject in need of imaging an inflammatory site.

13. Use of a carbonic apatite containing no contrast agent for producing an aggregation promoter for an inflammatory site of a contrast agent.

14. A carbonic apatite containing no contrast agent, which is used for a treatment for promoting aggregation of a contrast agent to an inflammatory site.