Calcium phosphate powder composition

Abstract

The present invention relates to a powder composition that can be used as an adhesive when mixed with water. The composition is based on calcium phosphate, phosphorylated amino acids, and acidic compounds.

Description

Technical Field

[0001] The present invention relates to a powder composition of a calcium phosphate compound, a phosphorylated amino acid, and an acidic compound. The powder composition can be used as an adhesive. Furthermore, the present invention relates to the use of the powder composition and a method of using the same.

Background Art

[0002] Calcium phosphate (CaP), particularly hydroxyapatite (Ca , ,

[0005] (PO4)6(OH)2, HA) is a mineral widely used in medical applications due to its similarity to the mineral components of bone and teeth and its biocompatibility. Furthermore, hydroxyapatite is non-toxic, biocompatible, and bioactive. This means that hydroxyapatite is not harmful and is not recognized as a foreign substance, while on the other hand, it can have a beneficial effect on bone remodeling. Therefore, hydroxyapatite is widely used in bone repair and also as a drug / gene delivery medium, catalyst, ion adsorption / exchange agent, optoelectronic material reagent, etc. Hydroxyapatite can be formed, for example, by mixing tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD) under basic conditions.

[0003] However, a problem with HA is its crystallinity and very high stability, which can make it difficult for the body to reconstruct the material so as to incorporate it into soft tissues, and also requires a very long time in the process.

[0004] The field of biomaterials includes tissue fixation and tissue repair of implants. The limit of the mechanical strength of implants using composite adhesives remains an issue in the field of implants and biomaterials. The repair of soft tissues and internal organs with adhesives has generally not been successful.

[0005] Patent document 1 (US'446) discloses adhesives containing polyvalent metal compounds, compounds containing serine phosphate oligomers, or polymers encapsulated with serine phosphate, the latter of which are present in 10-90% by weight. US'446 discloses experimental data using tetracalcium phosphate (TTCP) as the polyvalent metal compound, and, as an example, phosphoserine-ethylene glycol-diglycidyl-phosphoserine, achieving an adhesive strength of up to 3.76 MPa when bonded to bone.

[0006] Patent document 2 (US'057) discloses a bone repair composition comprising a bioactive glass material containing amino acid phosphate species, polyvalent metal compounds, and ionic functional groups. US'057 discloses an example using a composition containing TTCP, serine phosphate, and various amounts of Combeite Bioactive glass and water as the polyvalent metal compound, and bonding it to bone. The resulting shear strength was in the range of 0.75 to 2.13 MPa.

[0007] Patent Document 3 (US'189) teaches an adhesive composition having a polyvalent metal compound containing 10-90% by weight of a serine phosphate-like compound. US'189 discloses that when TTCP and various phosphorylated compounds are used as the polyvalent metal compound, the adhesive shear strength to cortical bone is 130-890 kPa after 5 minutes, and when α-TCP and phosphoserine are used, it is 650 kPa.

[0008] Several tissue adhesives are currently available on the market, but none are ideal sealants or adhesives. Cyanoacrylates exhibit good adhesion, but have been shown to cause inflammatory reactions during their degradation process. Fibrin adhesives have low adhesive strength but excellent biocompatibility. Other adhesives have drawbacks such as high cost, long curing times, or difficulty in adjusting curing time according to the tissue and situation. Soft tissue adhesives typically contain fibrin, gelatin, and various polysaccharides.

[0009] Conventional tissue adhesives are currently unable to withstand any significant shear force, and none have been shown or suggested to be effective for soft tissues. Therefore, there is a need for a composition that can be used as both a hard tissue adhesive and a soft tissue adhesive that provides high shear strength. [Prior art documents] [Patent Documents]

[0010] [Patent Document 1] U.S. Patent Application Publication No. 2012 / 288446 [Patent Document 2] U.S. Patent Application Publication No. 2013 / 0122057 [Patent Document 3] U.S. Patent No. 8765189 [Overview of the project]

[0011] The object of the present invention is to overcome the shortcomings of the prior art. Accordingly, in a first embodiment, the present invention relates to the composition described in claim 1.

[0012] In a second embodiment, the present invention relates to a method for producing stable amorphous calcium phosphate (ACP): a. A step of preparing a powder composition of a calcium phosphate compound selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP), a phosphorylated amino acid, and an acidic compound selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts, polyphosphates or polyphosphates or its salts; b. A step of adding an aqueous solution to the powder composition to obtain an aqueous composition; c. A step of measuring the pH of the aqueous composition; d. Optionally, the step of adjusting the pH to obtain a pH equal to or less than 9 by adding more calcium phosphate compounds, phosphorylated amino acids and / or acidic phosphate compounds. Regarding methods including

[0013] In a third aspect, the present invention relates to an aqueous composition comprising a powder composition and an aqueous solution according to the present invention.

[0014] In a fourth embodiment, the present invention relates to the use of an aqueous composition according to the present invention as a tissue adhesive.

[0015] In a fifth embodiment, the present invention relates to a method for adhering a first tissue to a second tissue using a powder composition according to the present invention: a. A step of obtaining a tissue adhesive by mixing the powder composition according to claim 1 with an appropriate amount of aqueous solution; b. Applying the tissue adhesive to the first or second tissue and optionally leaving it for an appropriate amount of time; c. The process of bringing the first tissue and the second tissue into contact with each other; d. Optionally, a step of applying appropriate time pressure to the first and second tissues; and e. Process for curing the tissue adhesive Regarding methods including

[0016] In a sixth aspect, the present invention relates to a kit comprising at least two containers, Any one container in the kit may contain one of the following: a calcium phosphate compound selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP) or a combination thereof; a phosphorylated amino acid; and an acidic compound selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts, polyphosphate or polyphosphate or its salts; and an aqueous solution. However, if one container contains a calcium phosphate compound, a phosphorylated amino acid, or an acidic compound, the aqueous solution must be contained in a separate container.

[0017] In a seventh aspect, the present invention is a composition for use as a tissue adhesive, the composition comprising a calcium phosphate compound selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP), a phosphorylated amino acid or creatine phosphate, and an acidic compound selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphoric acid or its salts, polyphosphates or polyphosphoric acid and its salts.

[0018] Embodiments by item In one embodiment of any of the aspects of the present invention, the amount of the phosphorylated amino acid or creatine phosphate is 10 to 80 mol%, preferably 10 to 60 mol%, more preferably 20 to 50 mol% based on the total dry matter.

[0019] The powder composition contains a total amount of calcium phosphate compound and acidic compound of 40 to 90 mol%, preferably 50 to 80 mol% based on the total dry matter.

[0020] In one embodiment of any of the aspects of the present invention, the molar% ratio in the total amount of the calcium phosphate compound and the acidic compound is 10 to 90:9 to 10.

[0021] In one embodiment, the amount of the calcium phosphate compound is preferably 20 to 50% by weight, more preferably 25 to 40% by weight based on the dry matter, and is preferably TTCP or α-TCP.

[0022] In one embodiment of any of the aspects of the present invention, the calcium phosphate compound is tetracalcium phosphate or α-TCP.

[0023] In one embodiment of any of the aspects of the present invention, the acidic compound is selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), and is preferably DCPD or DCPA.

[0024] In any embodiment of the present invention, the phosphorylated amino acid is phosphoserine, phosphothreonine, or phosphotyrosine.

[0025] In any embodiment of the present invention, the amount of the acidic compound is 20 to 50% by weight on a total dry weight basis.

[0026] In any embodiment of the present invention, the composition comprises 25 to 60 mol%, preferably 30 to 50 mol%, of phosphorylated amino acids or creatine phosphate, and a total amount of 40 to 75 mol%, preferably 50 to 70 mol%, of calcium phosphate compounds and acidic compounds, wherein the mol% ratio of the total amount of calcium phosphate compounds and acidic compounds is 60 to 90:40 to 10, preferably 65 to 90:53 to 10.

[0027] In any embodiment of the present invention, the composition comprises 15 to 40 mol%, preferably 20 to 30 mol%, of phosphorylated amino acids or creatine phosphate, and a total amount of 60 to 85 mol%, preferably 70 to 80 mol%, of calcium phosphate compounds and acidic compounds, wherein the molar ratio of the total amount of calcium phosphate compounds and acidic compounds is 10 to 50:90 to 50, preferably 10 to 40:90 to 60.

[0028] In any embodiment of the present invention, the composition further comprises a silicate compound, preferably in an amount of 10 to 30 mol% on a total dry weight basis.

[0029] In any embodiment of the present invention, the silicate compound is calcium silicate, preferably mono, di, or tricalcium silicate compounds, or a mixture of two or more, preferably calcium metasilicate.

[0030] In any embodiment of the present invention, the composition is essentially water-free, for example, less than 0.5% by weight, preferably less than 0.1% by weight.

[0031] In any embodiment of the present invention, the aqueous composition has a pH equal to or less than 9, preferably equal to or less than 8, more preferably equal to or less than 7, even more preferably equal to or less than 6, even more preferably equal to or less than 5, but preferably 3 or higher.

[0032] In any embodiment of the present invention, the pH of the composition is adjusted to be equal to or less than 8, more preferably equal to or less than 7, more preferably equal to or lower than 6, and more preferably equal to or less than 5.

[0033] In any embodiment of the present invention, the amount of aqueous solution is 15 to 50% by weight on a total weight basis.

[0034] In any embodiment of the present invention, at least one of the first tissue and the second tissue is soft tissue, preferably selected from tendons, ligaments, fascia, skin, fibrous tissue, muscle, fat, nerves, or blood vessels.

[0035] In any embodiment of the present invention, at least one of the first tissue and the second tissue is a hard tissue, preferably selected from bone or teeth.

[0036] In any embodiment of the present invention, the process is carried out in vitro.

[0037] In one embodiment, the calcium phosphate compound is tetracalcium phosphate (TTCP), the phosphorylated amino acid is phosphoserine, and the acidic compound is dicalcium phosphate dihydrate (DCPD).

[0038] All embodiments described herein can be combined with respect to all aspects of the present invention, unless otherwise specified. [Brief explanation of the drawing]

[0039] [Figure 1] This figure shows the shear strength of samples with different amounts of fixed phosphoserine and varying TTCP:DCPD molar ratios after curing at 22°C for 3 hours. [Figure 2] This graph shows the shear value (N) after 16 hours of curing. The 100% TTCP sample on the y-axis was cured for 24 hours. [Modes for carrying out the invention]

[0040] In this application, the term “aqueous solution” includes water and water of any purity. Water may be, but is not limited to, tap water, distilled water, or deionized water. The aqueous solution may be a buffer such as PBS, or any suitable serine buffer.

[0041] powder composition The composition according to the present invention is a powder composition comprising a calcium phosphate compound selected from tetracalcium phosphate (TTCP), tricalcium phosphate (TCP), or a combination of TTCP and TCP, a phosphorylated amino acid or creatine phosphate, and an acidic compound. The acidic compound is selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts, and polyphosphates or polyphosphates or its salts.

[0042] The inventors have discovered that this powder composition, when mixed with water, enables the formation and storage of amorphous calcium phosphate (ACP), which exhibits remarkably excellent mechanical strength when used, and is easy to handle when bonding tissues or other surfaces. In particular, this powder composition has been shown to be suitable for bonding hard tissue to hard tissue, soft tissue to hard tissue, or synthetic implants or scaffolds, as well as for bonding soft tissue to soft tissue. Furthermore, this composition is also suitable for use as a general-purpose adhesive for bonding surfaces together.

[0043] Considering the shear strength exhibited by TTCP and the fact that DCPD (acidic calcium phosphate) alone exhibits no shear strength at all, it is surprising that the mechanical properties were improved so significantly when these two components (TTCP and DCPD) were combined in the presence of phosphoserine (phosphorylated amino acid), as shown in Figure 1.

[0044] The inventors discovered that when the powder composition is mixed with water, the resulting aqueous composition not only forms ACP, but that this ACP maintains stability and is not converted to crystalline hydroxyapatite. Although not bound by theory, this unexpected effect is thought to be due to the combined effect of low pH and phosphorylated compounds (e.g., surface bonding of calcium salt particles or grains, ion chelation, increased solubility or dissolution, stabilization / destabilization of nucleation during cementation or crystallization processes), which promote the formation and stabilization of ACP. In one embodiment, the pH of the aqueous composition is preferably equal to or less than 9, more preferably equal to or less than 8, more preferably equal to or less than 7, more preferably equal to or less than 6, and more preferably equal to or less than 5. The pH of the composition is determined by preparing a sample containing 0.15 to 0.2 g of total dry powder, adding 0.25 mL / g of liquid, allowing it to harden for 5 minutes, then immersing the sample in 1 mL of water and measuring the pH of the water using any suitable pH measuring device or electrode until it equilibrates to a stable pH value.

[0045] The formed ACP is stable in an aqueous composition, in a wet or liquid environment, at room temperature for at least 1 day, preferably at least 3 days, more preferably at least 5 days, and more preferably at least 10 days. Stability is determined by measuring X-ray diffraction (XRD) and / or nuclear magnetic resonance (NMR), both of which indicate the proportion of crystalline and amorphous (ACP) solids.

[0046] The amount of calcium phosphate compound is preferably 20-50% by weight, more preferably 25-40% by weight, based on dry weight. In one embodiment, calcium phosphate is TTCP or α-TCP.

[0047] Surprisingly, the inventors discovered that, with respect to calcium and phosphate salts, the improvement effect of this composition applies only to tetracalcium phosphate and tricalcium phosphate. The calcium phosphate is preferably in the form of a powder having an average particle size of 5 to 5000 nm (for example, 20 nm or greater than 20 nm, 50 nm or greater than 50 nm, 100 nm or greater than 100 nm, 300 nm or greater than 300 nm, 500 nm or greater than 500 nm, 800 nm or greater than 800 nm, less than 3000 nm or less than 3000 nm, less than 1500 nm or less than 1500 nm, or less than 1000 nm or less). The particles may be spherical or flake-shaped.

[0048] Phosphorylation is the process by which a phosphate group (PO4) is added to an amino acid or other molecule. 3- This is the addition of ). The phosphorylated amino acid according to the present invention is, for example, phosphorylated serine, threonine, or tyrosine, but may be other amino acids. In one preferred embodiment, the phosphorylated amino acid is phosphorylated serine, also known as phosphoserine (pSer). The phosphorylated amino acid according to the present invention may be a monomer, a dimer, or a trimer.

[0049] Phosphorylated amino acids or creatine phosphate are thought to act as curing agents that give the composition improved mechanical strength. The amount of phosphorylated amino acids or creatine phosphate should be 10 to 80 mol% on a dry weight basis of the powder composition. To equilibrate the pH of the aqueous composition when the powder composition is mixed with water, the amount of phosphorylated amino acids or creatine phosphate can be varied according to the desired pH and amount of acidic compounds. In one embodiment, a preferred amount of phosphorylated amino acids or creatine phosphate is 10 to 60 mol% relative to the solid content, for example, 12 mol% or more than 12 mol%, 15 mol% or more than 15 mol%, 18 mol% or more than 18 mol%, 35 mol% or less than 35 mol%, 32 mol% or less than 32 mol%, 30 mol% or less than 30 mol%, 28 mol% or less than 28 mol%, and 25 mol% or less than 25 mol%. In one embodiment, the amount of phosphorylated amino acids or creatine phosphate is 20 to 40 mol%, preferably 15 to 30 mol%, on a dry weight basis. The advantage of using this amount is that the mechanical properties of the resulting adhesive are improved.

[0050] The acidic compound is selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts, and polyphosphates or polyphosphates or its salts. The polyphosphate may be potassium polyphosphate. In preferred embodiments, the acidic compound is DCPD or DCPA, preferably DCPD. These acidic compounds provide sufficient acidity and contribute to the mechanical properties of the adhesive composition when mixed with water.

[0051] The acidic compounds and calcium phosphate compounds are present in the powder composition in total amounts. In preferred embodiments, the total amount is 40 to 90 mol%, preferably 50 to 80 mol%, based on dry matter. Within this range, high shear strength was achieved when the powder mixture was mixed with water.

[0052] The molar ratio (mol% ratio) of the calcium phosphate compound and the acidic compound is preferably 10-90:90-10. The inventors have confirmed that the shear strength depends on this molar ratio depending on the amount of phosphorylated amino acid or creatine phosphate. When using a moderate amount of phosphorylated amino acid or creatine phosphate, the amount of calcium phosphate in the total amount should be higher, and when using a low amount, the amount of calcium phosphate should be lower. In one embodiment, the composition contains 25-60 mol%, preferably 30-50 mol%, of phosphorylated amino acid or creatine phosphate and a total amount of 40-75 mol%, preferably 50-70 mol%, of the calcium phosphate compound and the acidic compound, and the molar ratio of the total amount of the calcium phosphate compound and the acidic compound is 60-90:40-10, preferably 65-90:53-10. In another embodiment, the composition comprises 15-40 mol%, preferably 20-30 mol%, of phosphorylated amino acids or creatine phosphate, and a total amount of 60-85 mol%, preferably 70-80 mol%, of calcium phosphate compounds and acidic compounds, wherein the mol% ratio of the total amount of calcium phosphate compounds and acidic compounds is 10-50:90-50, preferably 10-40:90-60.

[0053] A preferred powder composition comprises phosphoserine, TTCP, and an acidic compound, preferably DCPD, wherein the composition contains 15 to 35 mol% phosphoserine, and the molar ratio of TTCP to the acidic compound is 10 to 50:90 to 50, preferably 10 to 40:90 to 60.

[0054] The powder composition preferably contains a silicate compound. In one embodiment, the composition is essentially free of silicate compounds or completely free of silicate compounds. In another embodiment, the amount of silicate compound is 5 to 30 mol%. In one embodiment, the amount of silicate is 8 mol% or more than 8 mol%, or 12 mol% or more than 12 mol%, or 25 mol% or less than 25 mol%, or 20 mol% or less than 20 mol%. Silicates further improve mechanical properties.

[0055] The silicate compound can be any suitable compound containing silicates, i.e., anionic silicon compounds. The silicon compound is [SiO4] 2- , or [Si2O7] 6- The silicate compound may be an oxide such as quartz, feldspar, zeolite, mica, pyroxene, etc. In one embodiment, the silicate compound is selected from calcium silicate, sodium silicate, and aluminum silicate, magnesium silicate, strontium silicate; zirconium silicate; or a mixture of dicalcium silicate and tricalcium silicate, preferably calcium silicate. The silicate may be in the form of cement, such as Portland gray cement or Portland white cement. The mixture of dicalcium silicate and tricalcium silicate may include 0 to 100% by weight of dicalcium silicate and 0 to 100% by weight of tricalcium silicate, for example, 30 to 70% by weight of dicalcium silicate and 30 to 70% by weight of tricalcium silicate. In one embodiment, the silicate compound is a monocalcium silicate compound, preferably calcium metasilicate.

[0056] The powder composition preferably contains virtually no water. That is, the amount of water on a total weight basis is preferably less than 0.5% by weight, and more preferably less than 0.1% by weight.

[0057] An advantage of the present invention is that the curing time can be adjusted according to the application so that curing occurs at the appropriate time. In some cases, the composition needs to cure rapidly, while in other cases, curing needs to be delayed because adjustments are required during application after mixing or molding for a while.

[0058] Retardants such as sodium citrate can be added to the reaction mixture in amounts of 0.1 to 10% by weight (based on solid content). In one embodiment, the amount of retarder is 3.5 to 7% by weight of the solid content. The retarder may be, but is not limited to, sodium citrate (monosodium citrate, disodium citrate, or trisodium citrate) or citric acid. The composition may further contain additives such as growth factors, nutrients, and antioxidants. However, the composition functions without retardants or additives, and in one embodiment, at least 95% by weight, preferably at least 98% by weight, and more preferably at least 99% by weight of the solid content of the powder composition consists of calcium phosphate compounds, phosphorylated amino acids or creatine phosphate, and acidic compounds.

[0059] aqueous composition By mixing the powder composition according to the present invention with an aqueous solution, an aqueous composition can be obtained, which can then be applied to a surface for adhesion.

[0060] The aqueous solution is preferably water, but may be a buffer such as PBS or any suitable salt buffer. The water in the aqueous solution may be, but is not limited to, tap water, distilled water, or deionized water.

[0061] The pH of the aqueous composition is important for obtaining the desired properties of the cured composition. In one preferred embodiment, the aqueous composition has a pH equal to or less than 9, preferably equal to or less than 8, more preferably equal to or less than 7, even more preferably equal to or less than 6, even more preferably equal to or less than 5, but preferably 3 or higher.

[0062] Based on the total weight of the aqueous composition, the composition contains an amount of aqueous solution such that the liquid-to-powder ratio (L / P) is 0.16 to 0.45, preferably 0.20 to 0.42 or 0.23 to 0.42. The powder refers to calcium phosphate and additive compounds, and the ratio is expressed as the ratio of volume (mL) to weight (g).

[0063] ACP manufacturing method To prepare the ACP according to the present invention, the powder composition is mixed with an aqueous solution, preferably water, but may also be a buffer such as PBS or any suitable salt buffer, by any suitable method. The water in the aqueous solution may be, but is not limited to, tap water, distilled water, or deionized water.

[0064] Amorphous calcium phosphate is formed after mixing the powder composition with an aqueous solution. The amount of aqueous solution added can be 20-50% by weight relative to the total weight of the resulting aqueous composition. The curing time, fluidity or moldability, and mechanical properties are affected by the amount of aqueous solution added. For example, a large amount of water reduces the adhesive strength. The water in the aqueous solution can be distilled water, deionized water, or any high-purity water, but tap water can also be used. The aqueous solution may also be in the form of a buffer or a hydrogel such as hyaluronic acid, polyvinyl alcohol, chitosan, collagen, or a combination thereof. Using a hydrogel as the aqueous solution makes it easier for the composition to remain in the desired position during curing.

[0065] Tissue adhesion method The aqueous composition according to the present invention can be used for a variety of applications. Due to the ease of application of the aqueous composition and the mechanical strength of the cured composition, this composition can be used as a tissue adhesive.

[0066] The tissue adhesive is formed by mixing the powder composition of the present invention with an aqueous solution. The aqueous solution is preferably water, a buffer solution, or a hydrogel. The water in the aqueous solution may be, but is not limited to, tap water, distilled water, or deionized water.

[0067] By applying tissue adhesive to tissue (hard or soft tissue), other tissue or artificial implants can be bonded, creating sufficient mechanical strength between the two surfaces.

[0068] To bond a first tissue to a second surface, a tissue adhesive is applied to the first tissue or the second surface. This could be, for example, two or more tissues, or tissue and an implant surface or scaffold. The tissue adhesive can also be applied to the second surface. The adhesive can be left for an appropriate amount of time before the two or more tissues or surfaces are brought into contact with each other. This time depends on the components and curing time of the adhesive, as well as the tissues and materials, but non-limiting examples include 10 seconds or longer, 30 seconds or longer, 1 minute or longer, or 5 minutes or longer. In one embodiment, the composition is left for 20 to 60 seconds before the two or more tissues or surfaces are brought into contact with each other.

[0069] Subsequently, the surfaces may be brought into contact and pressure may be applied as needed. The pressure applied depends on the curing time of the tissue / material and the tissue adhesive, but non-limiting examples include 10 seconds or longer, 30 seconds or longer, 1 minute or longer, or 5 minutes or longer. In one embodiment, pressure is applied for 1 to 3 minutes. To cure the tissue adhesive more quickly, energy may be applied to the composition or the tissue portion to which the composition is applied. This can be done by irradiating with ultraviolet light, heat, or any suitable type of radiation for several seconds to several minutes. The adhesive is then cured until it becomes the final cured composition. The adhesive is fully cured after 5 minutes to 48 hours, depending on the composition and the tissue or surface.

[0070] The curing time depends on the ratio of the specific components, but since the curing of the tissue adhesive begins when the powder composition is mixed with the aqueous solution, the mixing must be done at the right time. In certain applications, rapid curing after application of the tissue adhesive is required, while in other applications, gradual curing is desired. The present invention facilitates the adjustment of the curing time, enabling users to prepare the adhesive in advance so that it does not become completely cured at the time of application, to prepare a tissue adhesive that remains moldable, or to prepare a tissue adhesive that cures almost instantaneously.

[0071] This method can be performed in vivo or in vitro, although some steps can be performed in vitro followed by in vivo. Damage requiring extensive tissue replacement cannot heal without intervention. Currently, large tissue fragments for transplantation cannot be cultured because oxygen and nutrients cannot penetrate deeper than 500 μm to 2 mm. One possible solution to this problem is to culture multiple small tissue fragments, adhere them together (in vitro or ex vivo), and then join them using the composition of the present invention immediately before in vivo transplantation.

[0072] The tissue is soft tissue and may be selected from, but is not limited to, tendons, ligaments, cartilage, fascia, skin, fibrous tissue, muscle, fat, nerves, blood vessels, liver, stomach, hair, nails, eyelashes, intestines, bladder, brain, eyes, uterus, lungs, esophagus, heart, kidneys, spleen, and glands. In one embodiment, the soft tissue is selected from fascia, skin, fibrous tissue, muscle, fat, nerves, blood vessels, liver, stomach, intestines, bladder, brain, eyes, uterus, lungs, esophagus, heart, lungs, kidneys, spleen, and glands. In one embodiment, the soft tissue is cartilage or tendon. In one embodiment, the soft tissue adhesive according to the present invention is used to bond soft tissue to hard tissue or to bond two different soft tissues together. In one embodiment, the soft tissue is tissue having an extracellular matrix, collagen, and elastin. In another embodiment, the soft tissue is tissue having epithelium. The tissue may also be hard tissue preferably selected from bone or teeth.

[0073] Furthermore, the compositions according to the present invention can be used to bond implants or scaffolds to tissue. The implants or scaffolds may be made from synthetic materials, biological materials, or combinations thereof. Examples of synthetic materials include metals, polymers, or ceramics. Metals may be titanium, niobium, or alloys thereof, or aluminum oxide, stainless steel. Polymers may be polyurethane, polyester (e.g., polylactic acid, polyglycolic acid, polycaprolactone), polyacrylate (e.g., polymethyl methacrylate, poly(2-hydroxyethyl methacrylate)), polyether (e.g., polyethylene glycol), polysiloxane (e.g., silicone), hydrogel (e.g., polyvinyl alcohol), and polyvinyl (e.g., polyethylene, polypropylene, polyisobutylene, polystyrene). Ceramics may be calcium phosphate (e.g., hydroxyapatite, mononite, tetracalcium phosphate), metal oxides (e.g., aluminum oxide, zirconium oxide, titanium oxide), or bioglass.

[0074] Adhesive preparation kit A kit containing different components of the powder composition can be used for the use of this composition. The kit includes at least two containers, which can be any suitable type of container, such as a bowl, bag, dish, plate, beaker, flask, can, cup, or bottle, and can be of any size and shape. One of the containers in the kit may contain a calcium phosphate compound selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP), a phosphorylated amino acid or creatine phosphate, and dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts, and polyphosphates or polyphosphates or their salts, as well as an aqueous solution. However, if one of the containers contains a calcium phosphate compound, a phosphorylated amino acid or creatine phosphate, and an acidic compound, the aqueous solution must be contained in a separate container; otherwise, the composition will harden. In other words, one container may contain an aqueous solution, and a second or additional container may contain the solid components, i.e., a calcium phosphate compound, a phosphorylated amino acid or creatine phosphate, and an acidic compound. When silicate compounds are used, they may exist in the form of two or more reactants that can react to form the silicate compound. The two or more reactants may be contained in the same container or in separate compartments. In one embodiment, the kit includes three or more containers. The amounts of aqueous solutions, calcium compounds, phosphorylated amino acids or creatine phosphates, and acidic compounds in the kit are such that when mixed, an aqueous composition or tissue adhesive of the desired amounts and ratios is obtained. [Examples]

[0075] Example 1 Materials: Calcium silicate (CS1, Sigma-made calcium metasilicate), tetracalcium phosphate (TTCP, synthesized at Uppsala University), dicalcium phosphate dihydrate (DCPD, Sigma), α-tricalcium phosphate (aTCP, Innotere 5μm), and phosphoserine (Flamma) were used as reactants / starting materials.

[0076] Sample: The powder was prepared as shown in the table or graph, pre-mixed (total 0.2g), mixed with water (50μL, 0.25L / P), and polished with silicon carbide abrasive paper of 60 grit to form a steel cube (1cm). 3 The samples were applied to the surface of the material. Each sample was mixed within 15-20 seconds. Two cubes were pressed together using grips (Cocraft grips) along with the adhesive between them, and cured at room temperature (22°C, 40% relative humidity) for 3 hours, and some samples for 16 hours.

[0077] Testing: Samples were tested using a mechanical testing machine from Shimadzu (AGS-H, Shimadzu Europa GmbH, Duisburg, Germany) at a displacement rate of 1 mm / min and a 5 kN load cell, in a shear test setting. Data were analyzed using the manufacturer's software, Trapezium-X Lite, version 1.2.0 (Shimadzu Europa). Each group or data point represents the mean value for sample sizes N=2 to 4.

[0078] Results: The results are shown in Figure 1. Very high shear strength was already observed 3 hours after the start of curing. No significant increase in shear strength was observed even when the curing time was extended to 16 hours (Figure 2), indicating that this composition cures completely within 3 hours.

[0079] Several samples containing monocalcium silicate were prepared and tested in the same manner as described above. The results are shown in Table 1.

[0080] [Table 1]

Claims

1. A powder composition comprising: a calcium phosphate compound selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP) or a combination thereof; a phosphorylated amino acid or creatine phosphate; and an acidic compound selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts; and polyphosphates or polyphosphates or its salts.

2. The powder composition according to claim 1, wherein the amount of phosphorylated amino acids or creatine phosphate is 10 to 80 mol%, preferably 10 to 60 mol%, and more preferably 20 to 50 mol%, based on the total dry matter.

3. The powder composition according to claim 1 or 2, wherein the powder composition contains a total amount of calcium phosphate compound and acidic compound in an amount of 40 to 90 mol%, preferably 50 to 80 mol%, based on total dry matter.

4. The powder composition according to claim 3, wherein the molar percentage ratio of the total amount of the calcium phosphate compound and the acidic compound is 10 to 90:90 to 10.

5. The powder composition according to claim 1, wherein the calcium phosphate compound is tetracalcium phosphate or α-TCP, and the amount of the calcium phosphate compound is preferably 20 to 50% by weight, more preferably 25 to 40% by weight, on a dry weight basis.

6. The powder composition according to claim 1, wherein the acidic compound is selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), and monocalcium phosphate monohydrate (MCPM), preferably DCPD or DCPA.

7. The powder composition according to any one of claims 1 to 6, wherein the phosphorylated amino acid is phosphoserine, phosphothreonine, or phosphotyrosine.

8. The powder composition according to claim 8, wherein the amount of the acidic compound is 20 to 50% by weight on a total dry material basis.

9. The powder composition according to any one of claims 1 to 8, comprising 25 to 60 mol%, preferably 30 to 50 mol%, of phosphorylated amino acids or creatine phosphate, and a total amount of 40 to 75 mol%, preferably 50 to 70 mol%, of calcium phosphate compounds and acidic compounds, wherein the mol% ratio of the total amount of calcium phosphate compounds and acidic compounds is 60 to 90:40 to 10, preferably 65 to 90:53 to 10.

10. The powder composition according to any one of claims 1 to 8, comprising 15 to 40 mol%, preferably 20 to 30 mol%, of phosphorylated amino acids or creatine phosphate, and a total amount of 60 to 85 mol%, preferably 70 to 80 mol%, of calcium phosphate compounds and acidic compounds, wherein the mol% ratio of the total amount of calcium phosphate compounds and acidic compounds is 10 to 50:90 to 50, preferably 10 to 40:90 to 60.

11. The powder composition according to any one of claims 1 to 10, wherein the calcium phosphate compound is tetracalcium phosphate (TTCP), the phosphorylated amino acid is phosphoserine, and the acidic compound is dicalcium phosphate dihydrate (DCPD).

12. The powder composition according to claim 1, further comprising a silicate compound, preferably in an amount of 10 to 30 mol% on a total dry weight basis.

13. The powder composition according to claim 12, wherein the silicate compound is calcium silicate, preferably a mono, di, or tricalcium silicate compound, or a mixture of two or more, preferably calcium metasilicate.

14. The powder composition according to any one of claims 1 to 13, wherein the composition is essentially water-free, for example, less than 0.5% by weight, preferably less than 0.1% by weight.

15. An aqueous composition comprising the powder composition according to any one of claims 1 to 14 and an aqueous solution.

16. The aqueous composition according to claim 15, wherein the aqueous composition has a pH equal to or less than 9, preferably equal to or less than 8, more preferably equal to or less than 7, even more preferably equal to or less than 6, even more preferably equal to or less than 5, but preferably 3 or higher.

17. Use of the aqueous composition according to claim 15 or 16 as a tissue adhesive.

18. A method for producing stable amorphous calcium phosphate (ACP): a. A step of preparing powder compositions of calcium phosphate compounds selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP), phosphorylated amino acids, and acidic compounds selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts, polyphosphates or polyphosphates or its salts; b. A step of adding an aqueous solution to a powder composition to obtain an aqueous composition; c. A step of measuring the pH of the aqueous composition; d. Optionally, the step of adjusting the pH to obtain a pH equal to or less than 9 by adding more calcium phosphate compounds, phosphorylated amino acids, and / or acidic phosphate compounds. A method that includes this.

19. The pH of the composition is equal to or less than 8, more preferably equal to or less than 7, more preferably equal to or less than 6, more preferably equal to or less than 5, the method according to claim 18.

20. The method according to claim 18 or 19, wherein the amount of aqueous solution is 15 to 50% by weight on a total weight basis.

21. A method for adhering a first tissue to a second tissue using the powder composition described in claim 1, the method being: a. A step of obtaining a tissue adhesive by mixing the powder composition according to claim 1 with an appropriate amount of aqueous solution; b. The process of applying the tissue adhesive to the first or second tissue and optionally leaving it for an appropriate amount of time; c. The process of bringing the first tissue and the second tissue into contact with each other; d. Optionally, a step of applying appropriate time pressure to the first and second tissues; and e. Process for curing the tissue adhesive A method that includes this.

22. The method according to claim 21, wherein at least one of the first tissue and the second tissue is soft tissue, preferably selected from tendons, ligaments, fascia, skin, fibrous tissue, muscle, fat, nerves or blood vessels.

23. The method according to claim 21, wherein at least one of the first tissue and the second tissue is a hard tissue, preferably selected from bone or tooth.

24. The method according to any one of claims 21 to 23, wherein the process is performed in vitro.

25. A kit comprising at least two containers, Any one container in the kit may contain one of the following: a calcium phosphate compound selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP) or a combination thereof, a phosphorylated amino acid, and an acidic compound selected from dicalcium phosphate dihydrate (DCPD), dicalcium phosphate anhydride (DCPA), monocalcium phosphate monohydrate (MCPM), pyrophosphate or its salts, polyphosphate or polyphosphate or its salts, or an aqueous solution; however, if one container contains a calcium phosphate compound, a phosphorylated amino acid, or an acidic compound, the aqueous solution must be contained in a separate container.

26. A composition for use as a tissue adhesive, comprising a calcium phosphate compound selected from tetracalcium phosphate (TTCP) or tricalcium phosphate (TCP), a phosphorylated amino acid or creatine phosphate, and an acidic compound selected from dicalcium phosphate dihydrate (DCPD), anhydrous dicalcium phosphate (DCPA), monohydrate monocalcium phosphate (MCPM), pyrophosphate or its salts, polyphosphate or polyphosphate or its salts.

Images