Method using predetermined quantitative value of collagen contained in mammalian teeth

Abstract

To provide a method using a predetermined quantitative value of collagen contained in mammalian teeth as an indicator for evaluating the potential risk of disease in a mammal and / or whether a mammal is potentially healthy or not.SOLUTION: At least one of PYD, PEN, T-GLYC, PpP, and PpG is compared with respective reference values thereof with respect to each of collagen contained in the teeth of a mammal to be evaluated for a potential risk of developing a disease and / or whether the mammal is potentially healthy and collagen contained in the teeth of one or more mammals selected from a mammal that has not developed a disease, a mammal that has developed a disease, a healthy mammal, and a non-healthy mammal.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a method using a quantitative value of the amount of pyridinoline contained in collagen (ng / mg) in the teeth of a mammal, a method using a quantitative value of the amount of pentosidine contained in collagen (ng / mg) in the teeth of a mammal, a method using a quantitative value of the total amount of glycated products contained in collagen (ng / mg: converted to the amount of pentosidine contained) in the teeth of a mammal, a method using a value of the weight ratio (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)) calculated from the quantitative value of the amount of pyridinoline contained in collagen (ng / mg) and the quantitative value of the amount of pentosidine contained (ng / mg) in the teeth of a mammal, and a method using a value of the weight ratio (amount of pyridinoline contained (ng / mg) / total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained)) calculated from the quantitative value of the amount of pyridinoline contained in collagen (ng / mg) and the quantitative value of the total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained) in the teeth of a mammal, relating to at least any one of the methods.

Background Art

[0002] Conventionally, various collagens (actually, "collagen-rich organic compositions" are accurate, but generally, they are often simply referred to as "collagen") have been developed. Most of them are atelocollagens produced by being extracted from soft tissues in the living body using an enzyme treatment method, in which telopeptides present at the N-terminal and C-terminal in the amino acid sequence of collagen are cleaved, and insoluble collagen fibers are excluded during the purification process. Also, collagens extracted and produced from hard tissues such as bone and teeth, such as demineralized freeze-dried bone allograft (DFDBA) and demineralized dentin matrix (DDM), have been developed. Collagen is produced by demineralization treatment and is mainly used as a biomaterial.

[0003] Regarding atelocollagen, the quality of collagen is evaluated because telopeptides, which are said to have antigenicity, are cleaved by enzymatic treatment. For non-denatured collagen derived from soft tissues (collagen having a triple helix structure with telopeptides retained), the quality of collagen is evaluated in terms of how highly purified collagen is produced by setting the conditions for purification processes such as pH. By the way, when producing collagen from hard tissues, it is necessary to remove minerals by decalcification treatment, i.e., acid treatment, and then extract collagen. Moreover, the yield of collagen is limited and the extraction is time-consuming. Therefore, almost all of the developed collagen products are atelocollagen, non-denatured collagen derived from soft tissues, or products using them.

[0004] Most of the collagen contained in biological tissues is type I collagen. Type I collagen has both physiological and non-physiological crosslinks. Pyridinoline crosslink molecules are known as physiological crosslink molecules, and crosslink molecules formed by advanced glycation end products (AGEs) represented by pentosidine are known as non-physiological crosslink molecules (Non-Patent Document 1). The pyridinoline crosslink is a physiological and enzymatic crosslink due to the action of lysyl oxidase and is a regular crosslink molecule at specific sites in the amino acid sequence of collagen, contributing to the improvement of the elasticity and strength of collagen fibers. On the other hand, the pentosidine crosslink is a non-physiological and non-enzymatic crosslink (aging crosslink (AGE crosslink)) involving blood glucose. When blood glucose is warmed by body temperature, it is a crosslink molecule formed between arginine and lysine scattered in the amino acid sequence of collagen. Therefore, it is a randomly formed crosslink molecule between adjacent arginine and lysine in the amino acid sequence of collagen, which disrupts the physiological three-dimensional structure of collagen and contributes to the decrease in the elasticity and strength of collagen fibers. As described above, atelocollagen has its telopeptides cleaved by enzymatic treatment, so it contains almost no pyridinoline crosslink molecules. Moreover, during the purification process, insoluble collagen fibers caused by pentosidine crosslinks are excluded, so it is also collagen that contains almost no pentosidine crosslink molecules.

[0005] Conventionally, it is known that an increase in pyridinoline cross-linked molecules in collagen increases the strength and elasticity of the collagen (Non-Patent Document 2). In addition, it is known that pentosidine cross-linking is a pathological cross-linking (aging cross-linking (AGEs cross-linking)) (Non-Patent Document 3). Further, it has been reported that the amount of pentosidine per unit collagen in human articular cartilage increases linearly with age, and the increase in the ratio of the amount of pentosidine to the amount of pyridinoline per unit collagen (content of pentosidine / content of pyridinoline) accelerates with age (Non-Patent Documents 4 and 5). There is also a report that in non-calcified lesions of dystrophic calcification of the aorta, the ratio of the content of pentosidine to the content of pyridinoline per unit collagen (content of pentosidine / content of pyridinoline) increases significantly (Non-Patent Document 6).

Prior Art Documents

Non-Patent Documents

[0006]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Summary of the Invention

Problems to be Solved by the Invention

[0007] The evaluation of the quality of the above-mentioned collagen (collagen-rich organic composition) is generally evaluated based on the level of purity and nature of collagen in the collagen-rich organic composition, and not based on any molecular-level index of collagen, that is, any index at the amino acid sequence level of collagen. Therefore, it is difficult to say that the quality of collagen itself has been correctly evaluated. In addition, the amount of pyridinoline contained, which contributes to the improvement of the quality of collagen such as the elasticity and strength of collagen fibers, and factors that disrupt the physiological three-dimensional structure of collagen, cause a decrease in the elasticity and strength of collagen fibers, and ultimately contribute to a decrease in the quality of collagen. The amount of pentosidine contained and the total amount of glycated products, the weight ratio of the amount of pyridinoline contained and the amount of pentosidine contained, and the weight ratio of the amount of pyridinoline contained and the total amount of glycated products have not been used to evaluate the quality of collagen.

[0008] In addition, in Non-Patent Documents 1 to 6 described above, although there are various mentions of pyridinoline cross-linked molecules and pentosidine cross-linked molecules in collagen, the ratio of the amount of pentosidine contained per unit collagen to the amount of pyridinoline contained (amount of pentosidine contained / amount of pyridinoline contained) in diseases has been considered. However, none of the documents are specifically about collagen derived from teeth, where collagen extraction is not easy. Also, in tissues that require physiological strength, such as hard tissues like bone and cartilage and soft tissues like tendons and ligaments, the amount of pyridinoline cross-linked molecules contained in the collagen is large, and accordingly, the amount of pentosidine cross-linked molecules or cross-linked molecules of total glycation products tends to be large (M. TAKAHASHI et al., Anal Biochem Vol. 232, 158 - 62, 1995). However, since these tissues are metabolized tissues, the pyridinoline cross-linked molecules, pentosidine cross-linked molecules, and cross-linked molecules of total glycation products contained therein are also metabolized, so it does not reflect the total accumulated amount of pyridinoline cross-linked molecules, pentosidine cross-linked molecules, and cross-linked molecules of total glycation products accumulated in the mammalian host so far. On the other hand, teeth, which are hard tissues, are the only tissues among hard tissues that are not incorporated into the metabolic cycle. Therefore, the pyridinoline cross-linked molecules, pentosidine cross-linked molecules, and cross-linked molecules of total glycation products contained in the collagen are not metabolized, and thus the total accumulated amount of pyridinoline cross-linked molecules, pentosidine cross-linked molecules, and cross-linked molecules of total glycation products accumulated in the mammalian host so far is reflected. Therefore, in order to accurately evaluate the potential risk of diseases in mammals and whether mammals are potentially healthy, it is obvious that collagen derived from teeth (collagen-rich organic composition) that reflects the total accumulation of pyridinoline cross-linked molecules, pentosidine cross-linked molecules, and cross-linked molecules of total glycation products accumulated in the mammalian host so far should be used as an indicator. However, in Non-Patent Documents 1 to 6 described above, there is no such description, let alone any suggestion.

[0009] The present invention is a method using a quantitative value of the content of pyridinoline in collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a disease in a mammal and / or whether the mammal is potentially healthy, the method comprising: quantifying the content of pyridinoline in collagen (ng / mg) contained in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, to obtain a quantitative value PYD; quantifying the content of pyridinoline in collagen (ng / mg) contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals, to obtain a reference quantitative value R-PYD; and comparing the PYD and the R-PYD. The present invention is also a method using a quantitative value of the content of pentosidine in collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a disease in a mammal and / or whether the mammal is potentially healthy, the method comprising: quantifying the content of pentosidine in collagen (ng / mg) contained in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, to obtain a quantitative value PEN; quantifying the content of pentosidine in collagen (ng / mg) contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals, to obtain a reference quantitative value R-PEN; and comparing the PEN and the R-PEN. The present invention is further a method using a quantitative value of the total glycation product content in collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a disease in a mammal and / or whether the mammal is potentially healthy, the method comprising: quantifying the total glycation product content in collagen (ng / mg: converted to pentosidine content) contained in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, to obtain a quantitative value T-GLYC; quantifying the total glycation product content in collagen (ng / mg: converted to pentosidine content) contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals,A method comprising a step of quantifying the total amount of glycated products contained in collagen in teeth (ng / mg: converted to the amount of contained pentosidine) to obtain R-T-GLYC which is a reference quantitative value, and a step of comparing the T-GLYC and the R-T-GLYC, and as an index for evaluating the potential risk of disease in a mammal and / or whether the mammal is potentially healthy, a method using a value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in collagen in the teeth of a mammal and the quantitative value of the amount of pentosidine contained therein, which comprises a step of quantifying the amount of pyridinoline contained in collagen in the teeth (ng / mg) of a mammal to be evaluated for the potential risk of disease and / or whether the mammal is potentially healthy to obtain PYD which is a quantitative value, a step of quantifying the amount of pentosidine contained in collagen in the teeth (ng / mg) of the mammal to be evaluated to obtain PEN which is a quantitative value, a step of calculating PpP which is a value of the weight ratio (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)) from the obtained PYD and the obtained PEN, a step of quantifying the amount of pyridinoline contained in collagen in the teeth (ng / mg) of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals and non-healthy mammals to obtain R-PYD which is a reference quantitative value, a step of quantifying the amount of pentosidine contained in collagen in the teeth (ng / mg) of the one or more mammals to obtain R-PEN which is a reference quantitative value, a step of calculating R-PpP which is a reference value of the weight ratio (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)) from the obtained R-PYD and the obtained R-PEN, and a step of comparing the PpP and the R-PpP, and as an index for evaluating the potential risk of disease in a mammal and / or whether the mammal is potentially healthy, a method using a value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in collagen in the teeth of a mammal and the quantitative value of the total amount of glycated products contained therein, which comprises a step of, for a mammal to be evaluated for the potential risk of disease and / or whether the mammal is potentially healthy,A step of quantifying the amount of pyridinoline contained in collagen in teeth (ng / mg) to obtain PYD which is a quantitative value, a step of quantifying the total amount of glycated products contained in collagen in the teeth of the mammal to be the subject of the evaluation (ng / mg: converted amount of contained pentosidine) to obtain T-GLYC which is a quantitative value, a step of calculating PpG which is a value of weight ratio (amount of contained pyridinoline (ng / mg) / total amount of glycated products contained (ng / mg: converted amount of contained pentosidine)) from the obtained PYD and the obtained T-GLYC, a step of quantifying the amount of pyridinoline contained in collagen in teeth (ng / mg) of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals and non-healthy mammals to obtain R-PYD which is a reference quantitative value, a step of quantifying the total amount of glycated products contained in collagen in the teeth of the one or more mammals (ng / mg: converted amount of contained pentosidine) to obtain R-T-GLYC which is a reference quantitative value, a step of calculating R-PpG which is a reference value of weight ratio (amount of contained pyridinoline (ng / mg) / total amount of glycated products contained (ng / mg: converted amount of contained pentosidine)) from the obtained R-PYD and the obtained R-T-GLYC, and a step of comparing the PpG and the R-PpG, with the aim of providing at least any one of the methods of the method including these steps.

Means for Solving the Problems

[0010] As a result of intensive studies to solve the above problems, the present inventors have found that by comparing at least any one of the above-described values of PYD, PEN, T-GLYC, PpP, and PpG for each of the collagen contained in the teeth of a mammal to be the subject of evaluation of the potential risk of suffering from a disease and / or whether the mammal is potentially healthy, and the respective reference values, it is possible to evaluate the potential risk of suffering from a disease in a mammal and / or whether the mammal is potentially healthy, and have completed the following inventions.

[0011] (1) A method using a quantitative value of the amount of pyridinoline contained in collagen in the teeth of a mammal as an indicator for evaluating the potential risk of a disease in a mammal and / or whether the mammal is potentially healthy, the method comprising: quantifying the amount of pyridinoline contained in collagen in the teeth (ng / mg) of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy to obtain a quantitative value PYD; quantifying the amount of pyridinoline contained in collagen in the teeth (ng / mg) of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain a reference quantitative value R-PYD; and comparing the PYD and the R-PYD.

[0012] (2) A method using a quantitative value of the amount of pentosidine contained in collagen in the teeth of a mammal as an indicator for evaluating the potential risk of a disease in a mammal and / or whether the mammal is potentially healthy, the method comprising: quantifying the amount of pentosidine contained in collagen in the teeth (ng / mg) of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy to obtain a quantitative value PEN; quantifying the amount of pentosidine contained in collagen in the teeth (ng / mg) of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain a reference quantitative value R-PEN; and comparing the PEN and the R-PEN.

[0013] (3) A method using the quantitative value of the total amount of glycated products of collagen contained in the teeth of a mammal as an indicator for evaluating the potential risk of disease in the mammal and / or whether the mammal is potentially healthy, the method comprising: quantifying the total amount of glycated products of collagen contained in the teeth of the mammal to be evaluated for the potential risk of disease and / or whether the mammal is potentially healthy (ng / mg: converted to the amount of contained pentosidine) to obtain a quantitative value of T-GLYC; quantifying the total amount of glycated products of collagen contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals (ng / mg: converted to the amount of contained pentosidine) to obtain a reference quantitative value of R-T-GLYC; and comparing the T-GLYC and the R-T-GLYC.

[0014] (4) As an index for evaluating the potential risk of a disease in a mammal and / or whether the mammal is potentially healthy, a method using a value of a weight ratio calculated from the quantitative value of the amount of pyridinoline contained in collagen in the teeth of the mammal and the quantitative value of the amount of pentosidine contained therein, comprising: a step of quantifying the amount of pyridinoline contained in collagen in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, and obtaining PYD as a quantitative value; a step of quantifying the amount of pentosidine contained in collagen in the teeth of the mammal to be evaluated, and obtaining PEN as a quantitative value; a step of calculating PpP (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), which is a value of a weight ratio, from the obtained PYD and the obtained PEN; a step of quantifying the amount of pyridinoline contained in collagen in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals, and obtaining R-PYD as a reference quantitative value; a step of quantifying the amount of pentosidine contained in collagen in the teeth of the one or more mammals, and obtaining R-PEN as a reference quantitative value; a step of calculating R-PpP (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), which is a reference value of a weight ratio, from the obtained R-PYD and the obtained R-PEN; and a step of comparing the PpP and the R-PpP.

[0015] (5) A method using a value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in collagen in the teeth of a mammal and the quantitative value of the total amount of glycated products contained therein as an index for evaluating the potential risk of disease in a mammal and / or whether the mammal is potentially healthy, the method comprising: a step of quantifying the amount of pyridinoline contained in collagen in the teeth of a mammal to be evaluated for the potential risk of disease and / or whether the mammal is potentially healthy to obtain a quantitative value PYD; a step of quantifying the total amount of glycated products contained in collagen in the teeth of the mammal to be evaluated to obtain a quantitative value T-GLYC (converted to the amount of pentosidine contained); a step of calculating PpG, which is a value of the weight ratio (amount of pyridinoline contained (ng / mg) / total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained)), from the obtained PYD and the obtained T-GLYC; a step of quantifying the amount of pyridinoline contained in collagen in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain a reference quantitative value R-PYD; a step of quantifying the total amount of glycated products contained in collagen in the teeth of the one or more mammals to obtain a reference quantitative value R-T-GLYC; a step of calculating R-PpG, which is a reference value of the weight ratio (amount of pyridinoline contained (ng / mg) / total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained)), from the obtained R-PYD and the obtained R-T-GLYC; and a step of comparing the PpG and the R-PpG.

[0016] (6) The method according to (1), wherein the quantification step is a step of quantifying using high performance liquid chromatography with fluorescence detector (HPLC-Flu) using heptafluorobutyric acid and formic acid as ion pair reagents.

[0017] (7) The method according to any one of (1) to (6), wherein the mammal is one or more mammals selected from the group consisting of humans, cows, horses, pigs, wild boars, sheep, deer, dogs, and cats. [Effect of the Invention]

[0018] According to the present invention, by comparing the values of at least any one of the above-mentioned PYD, PEN, T-GLYC, PpP, and PpG for the collagen contained in the teeth of a mammal that is the subject of evaluation of the potential risk of suffering from a disease and / or whether the mammal is potentially healthy, with the respective reference values, with the collagen contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals, it is possible to evaluate the potential risk of suffering from a disease in a mammal and / or whether the mammal is potentially healthy. [Brief Description of the Drawings]

[0019]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

BEST MODE FOR CARRYING OUT THE INVENTION

[0020] Hereinafter, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the amount of pyridinoline contained in the collagen in the teeth of the mammal (ng / mg), as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the amount of pentosidine contained in the collagen in the teeth of the mammal (ng / mg), as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the total amount of glycation products contained in the collagen in the teeth of the mammal (ng / mg: converted amount of pentosidine contained), as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of the weight ratio (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)) calculated from the quantitative value of the amount of pyridinoline contained in the collagen in the teeth of the mammal (ng / mg) and the quantitative value of the amount of pentosidine contained (ng / mg), and as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of the weight ratio (amount of pyridinoline contained (ng / mg) / total amount of glycation products contained (ng / mg: converted amount of pentosidine contained)) calculated from the quantitative value of the amount of pyridinoline contained in the collagen in the teeth of the mammal (ng / mg) and the quantitative value of the total amount of glycation products contained (ng / mg: converted amount of pentosidine contained) will be described in detail. In this specification, the numerical range represented by "~" or "from" means a range including the numerical values described before and after "~" or "from" as the lower limit value and the upper limit value.

[0021] In the present invention, the term "mammal" basically refers to an animal that reproduces sexually, and most existing species are viviparous and raise their offspring with milk. Such an animal is not particularly limited as long as it meets these criteria. Examples of such animals include humans; anthropoid apes such as orangutans, gorillas, chimpanzees, bonobos, and monkeys belonging to the family Hylobatidae; monkeys other than those belonging to the family Hylobatidae, as well as large mammals such as cows, horses, pigs, wild boars, sheep, goats, deer, serows, bears, seals, sea lions, walruses, whales, etc.; and small mammals such as dogs, cats, rabbits, mice, rats, weasels, martens, mongooses, etc. Humans, cows, horses, pigs, wild boars, sheep, deer, dogs, and cats can be mentioned as preferred mammals, humans, cows, horses, pigs, dogs, and cats can be mentioned as more preferred mammals, and humans, cows, dogs, and cats can be mentioned as most preferred mammals.

[0022] In the present invention, the term "potential risk of contracting a disease" does not refer to a state of having contracted a disease, but rather to a state in which the host's defense response against the disease is functioning to the maximum extent and there is a risk of just barely not contracting the disease, meaning a state in which it is almost impossible to confirm that the host has contracted the disease based on subjective symptoms, objective findings, or conventional general examination findings.

[0023] Also, in the present invention, the term "potentially healthy" refers to a state in which the subject will not reach an unhealthy state, meaning a state in which it is almost impossible to confirm that the subject is unhealthy based on subjective symptoms, objective findings, or conventional general examination findings. In the present invention, the term "not potentially healthy" refers to a state in which the subject will not reach a healthy state, meaning a state in which it is almost impossible to confirm that the subject is healthy based on subjective symptoms, objective findings, or conventional general examination findings.

[0024] Collagen is one of the proteins that form the framework of various tissues in the living body, mainly constituting various organs, blood vessels, nerves, skin, ligaments, tendons, bones, cartilage, and dentin of teeth in vertebrates, and is the main component of the extracellular matrix of multicellular animals. In the present invention, "collagen" mainly refers to type I collagen. Also, the "collagen" of the present invention is dental collagen. Since teeth are tissues that are subjected to a strong load such as biting force, unlike soft tissues, dental collagen is required to have an extremely high strength to withstand the biting force.

[0025] Hard tissue collagen can be determined to be derived from hard tissue by detecting, for example, BMP-2, BMP-4, BMP-7, and osteocalcin, and can be determined to be derived from teeth by detecting dentin sialoprotein (DSP), dentin glycoprotein (DGP), and dentin phosphoprotein (DPP).

[0026] In the present invention, the pyridinoline content, pentosidine content, and total glycation product content of collagen are, respectively, pyridinoline that constitutes pyridinoline cross-linking molecules, which are physiological cross-linking molecules in collagen, pentosidine that constitutes pentosidine cross-linking molecules, which are non-physiological cross-linking molecules in collagen, and total glycation products that constitute cross-linking molecules of total glycation products, which are non-physiological cross-linking molecules in collagen. In the present invention, the amount of pyridinoline content (ng / mg) of collagen, the amount of pentosidine content (ng / mg) of collagen, and the amount of total glycation product content (ng / mg: converted to pentosidine content) of collagen are each quantified to obtain the quantitative values of PYD, PEN, and T-GLYC. Also, by dividing the PYD by the PEN, the value of PpP (pyridinoline content (ng / mg) / pentosidine content (ng / mg)), which is the weight ratio value of the PYD and the PEN, and by dividing the PYD by the T-GLYC, the value of PpG (pyridinoline content (ng / mg) / total glycation product content (ng / mg: converted to pentosidine content)), which is the weight ratio value of the PYD and the T-GLYC, are each calculated.

[0027] It is no exaggeration to say that the pyridinoline crosslink, a physiological and enzymatic crosslinking molecule, is programmed mainly for improving the inherent strength of hard tissues. As described above, it is said to be a crosslinking molecule formed between two lysine residues in the telopeptides respectively present at the N-terminus and C-terminus in the amino acid sequence of collagen and one lysine residue in the amino acid sequence excluding both ends of collagen by the enzymatic action of lysyl oxidase. Since the pyridinoline crosslinking molecule formed between two lysine residues in the telopeptides respectively present at the N-terminus and C-terminus in the amino acid sequence of collagen is a crosslinking molecule between collagen fibers, it can be said that it greatly contributes to maintaining the three-dimensional structure of tropocollagen (the entire collagen sequence), which has a triple helix structure.

[0028] Therefore, when the amount of pyridinoline crosslinking molecules increases in the collagen contained in living tissues, the quality of the collagen contained in the living tissues becomes flexible and tenacious, the strength and degree of calcification of the collagen increase, and it can become a factor for improving the activity of the cells constituting the living tissues, that is, a factor for easily maintaining the homeostasis of an individual by increasing the activity of tissue stem cells involved in tissue repair. Therefore, the amount of pyridinoline contained is larger in collagen derived from teeth and bones than in soft tissue collagen typified by the skin, and the collagen derived from teeth and bones becomes flexible, high-strength, and highly elastic.

[0029] On the one hand, cross-linking by advanced glycation end products (AGEs), typified by pentosidine cross-linking molecules which are non-physiological cross-linking molecules and cross-linking molecules of total glycation products containing pentosidine cross-linking molecules, is pathological cross-linking (aging cross-linking (AGE cross-linking)) caused by blood glucose. As described above, when blood glucose is heated by body temperature, it is a cross-linking molecule formed by various combinations of arginine and lysine scattered in the amino acid sequence of collagen. When arginine and arginine, lysine and lysine, and arginine and lysine in the amino acid sequence of collagen are in proximity, the pentosidine cross-linking molecules and cross-linking molecules of total glycation products containing pentosidine cross-linking molecules are formed non-specifically (randomly). Therefore, the collagen fibers may become entangled, losing the regularity of the collagen fiber arrangement and disturbing the three-dimensional structure of tropocollagen. Thus, an increase in pentosidine cross-linking molecules and cross-linking molecules of total glycation products containing pentosidine cross-linking molecules in the amino acid sequence of collagen leads to a decrease in the flexibility, strength, and elasticity of collagen fibers. The formation of these pathological cross-linking molecules is said to depend on the host's dietary habits (carbohydrate intake habits) and increase in its accumulation amount because it is caused by blood glucose. Also, since carbohydrates, which are the raw materials for pentosidine cross-linking molecules and cross-linking molecules of total glycation products containing pentosidine cross-linking molecules, are supplied transvascularly, collagen, which is the support base of tissues throughout the body, is cross-linked according to the circulating blood flow volume regardless of the tissue location. That is, it can be said that excessive sugar intake leads to a uniform decrease in the quality of collagen, which is the support base of the body, regardless of hard or soft tissues.

[0030] In addition, when the cross-linked molecules of pentosidine and the total glycated products containing the cross-linked molecules of pentosidine increase in collagen, since the cross-linked molecules of pentosidine and the total glycated products containing the cross-linked molecules of pentosidine are glycated cross-linked molecules by the Maillard reaction mediated by sugar, the collagen contained in the living tissue exhibits a brittle chalk-like appearance, and the strength and degree of calcification of the collagen contained in the living tissue decrease. The products formed by the Maillard reaction mediated by sugar that have matured are called advanced glycation end products (AGEs). The cross-linked molecules of pentosidine and the total glycated products containing the cross-linked molecules of pentosidine are AGEs. In the collagen of mammals in which the blood sugar level is constantly maintained at a high level due to a diet with a high frequency of carbohydrate intake, compared with the collagen of mammals with a balanced diet of the same age (monthly age), since the cross-linked molecules of pentosidine and the total glycated products containing the cross-linked molecules of pentosidine are contained in a large amount, it can be said that pathological cross-linking (aging cross-linking (AGE cross-linking)) is progressing in the collagen of mammals with a constantly high blood sugar level. The generated cross-linked molecules of pentosidine and the total glycated products containing the cross-linked molecules of pentosidine disrupt the regular arrangement order of collagen fibers, impair the flexibility of collagen, change it into a hard and brittle substance, inhibit the metabolism of collagen, and as a result, deteriorate the quality of collagen that constitutes the whole body, causing a decrease in the flexibility of tissues. Since collagen is the main component of the supporting tissue that supports not only all hard tissues such as bones and teeth but also all soft tissues including nerves and blood vessels, this decrease in the quality of collagen can be a factor that deteriorates the quality of hard and soft tissues in the body. For example, when bone mass decreases due to bone glycation, the bone becomes brittle and osteoporosis occurs. In addition, when the quality of blood vessels decreases due to glycation of the blood vessel wall, the arteries become hardened, so-called arteriosclerosis occurs, which causes tissue damage such as an increase in blood pressure.The same is true for the three major complications of diabetes, namely, "(1) diabetic retinopathy (which may lead to blindness if it worsens)", "(2) diabetic nephropathy (which may require hemodialysis if it worsens)", and "(3) diabetic neuropathy (which may lead to necrosis of the fingers and toes due to blood circulation disorders, for example, if it worsens)". The causes are respectively said to be "(1) blood circulation disorders in the retina", "(2) capillary disorders in the glomeruli of the kidneys, which are blood filtration devices", and "(3) blood flow disorders in the extremities due to capillary neuropathy and arteriosclerosis". It can be said that in all these cases, the deterioration of blood vessel quality caused by the glycation of collagen that constitutes blood vessels is a major factor.

[0031] In addition, when advanced glycation end products (AGEs), represented by pentosidine cross-linked molecules and cross-linked molecules of total glycation products containing pentosidine cross-linked molecules, increase, AGEs bind to receptor for advanced glycation end products (RAGE), which is expressed in a wide range of tissues such as vascular cells represented by vascular endothelial cells and macrophages, which are immune cells. As a result, a biological environment in which its signal is easily activated is established. This promotes signals such as the NF-κB signal related to inflammation. AGEs, represented by pentosidine cross-linked molecules and cross-linked molecules of total glycation products containing pentosidine cross-linked molecules, function as chronic inflammation trigger factors for overly glycated tissues, thereby causing chronic inflammation in a wide range of tissues. This mechanism is considered to be a factor that exacerbates chronic inflammatory diseases such as inflammatory bowel disease (IBD) and collagen diseases. As described above, the glycation of collagen gradually erodes the living body in various ways, unknowingly reducing the quality of life (QOL) and becoming a major factor that distances organisms from healthy longevity. It should be noted that it has been reported that both pyridinoline cross-linked molecules and pentosidine cross-linked molecules in collagen contained in human biological tissues increase with aging (Shimizu, "Non-enzymatic glycation modification of dentin collagen protein related to aging", Osaka University Knowledge Archive. 2015; Walters C. et al., Calcif. Tissue Int. Vol. 35, 401-405, 1983).

[0032] As an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the quantitative value of the content of pyridinoline in collagen contained in the teeth of the mammal (ng / mg) is as follows: (i) Quantifying the content of pyridinoline in collagen contained in the teeth of the mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, and obtaining PYD, which is a quantitative value (step of quantifying the content of pyridinoline in the evaluation target); (ii) Quantifying the content of pyridinoline in collagen contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals, and obtaining R-PYD, which is a reference quantitative value (step of quantifying the content of pyridinoline in the reference); (iii) Comparing PYD obtained in the step of quantifying the content of pyridinoline in the evaluation target (i) and R-PYD obtained in the step of quantifying the content of pyridinoline in the reference (ii) (PYD·R-PYD comparison step); The above steps (i) to (iii) are included.

[0033] Also, as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the quantitative value of the content of pentosidine in collagen contained in the teeth of the mammal (ng / mg) is as follows: (iv) Quantifying the content of pentosidine in collagen contained in the teeth of the mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, and obtaining PEN, which is a quantitative value (step of quantifying the content of pentosidine in the evaluation target); (v) Quantifying the amount (ng / mg) of pentosidine contained in the collagen in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain R-PEN, which is a reference quantification value (reference pentosidine content quantification step), (vi) A step of comparing PEN obtained in the evaluation target pentosidine content quantification step (iv) and R-PEN obtained in the reference pentosidine content quantification step (v) (PEN·R-PEN comparison step), As described above, it has the steps of (iv) to (vi).

[0034] In addition, as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the quantification value of the total glycation product content (ng / mg: converted to pentosidine content) of the collagen contained in the teeth of the mammal is as follows: (vii) Quantifying the total glycation product content (ng / mg: converted to pentosidine content) of the collagen contained in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy to obtain T-GLYC, which is a quantification value (evaluation target total glycation product content quantification step), (viii) Quantifying the total glycation product content (ng / mg: converted to pentosidine content) of the collagen contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain R-T-GLYC, which is a reference quantification value (reference total glycation product content quantification step), (ix) A step of comparing T-GLYC obtained in the evaluation target total glycation product content quantification step (vii) and R-T-GLYC obtained in the reference total glycation product content quantification step (viii) (T-GLYC·R-T-GLYC comparison step), As described above, it has the steps of (vii) to (ix).

[0035] In addition, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of a weight ratio calculated from a quantitative value of the amount of pyridinoline contained in collagen in the teeth of the mammal and a quantitative value of the amount of pentosidine contained therein is (x) A step of quantifying the amount of pyridinoline contained in collagen (ng / mg) in the teeth of a mammal that is the subject of evaluation of the potential risk of disease and / or whether the mammal is potentially healthy to obtain PYD, which is a quantitative value (step of quantifying the amount of pyridinoline contained in the subject of evaluation), (xi) A step of quantifying the amount of pentosidine contained in collagen (ng / mg) in the teeth of the mammal that is the subject of the evaluation to obtain PEN, which is a quantitative value (step of quantifying the amount of pentosidine contained in the subject of evaluation), (xii) A step of calculating PpP, which is a value of the weight ratio (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), from PYD obtained in the step of quantifying the amount of pyridinoline contained in the subject of evaluation (x) and PEN obtained in the step of quantifying the amount of pentosidine contained in the subject of evaluation (xi) (step of calculating the weight ratio PpP of the subject of evaluation), (xiii) A step of quantifying the amount of pyridinoline contained in collagen (ng / mg) in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain R-PYD, which is a reference quantitative value (step of quantifying the amount of pyridinoline contained in the reference), (xiv) A step of quantifying the amount of pentosidine contained in collagen (ng / mg) in the teeth of the one or more mammals to obtain R-PEN, which is a reference quantitative value (step of quantifying the amount of pentosidine contained in the reference), (xv) A step of calculating R-PpP, which is a reference value of the weight ratio (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), from R-PYD obtained in the step of quantifying the amount of pyridinoline contained in the reference (xiii) and R-PEN obtained in the step of quantifying the amount of pentosidine contained in the reference (xiv) (step of calculating the reference weight ratio R-PpP), (xvi) Step of calculating the weight ratio PpP to be evaluated: A step of comparing PpP calculated in step (xii) and R-PpP calculated in the reference weight ratio R-PpP calculation step (xv) (weight ratio PpP·R-PpP comparison step). As described above, it has steps (x) to (xvi).

[0036] In addition, as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of a weight ratio calculated from a quantitative value of the amount of pyridinoline contained in collagen in the teeth of the mammal and a quantitative value of the total amount of glycated products contained is as follows. (xvii) Step of quantifying the amount of pyridinoline contained in collagen (ng / mg) in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy to obtain PYD as a quantitative value (step of quantifying the amount of pyridinoline contained in the evaluation target). (xviii) Step of quantifying the total amount of glycated products (ng / mg: converted to the amount of pentosidine contained) contained in collagen in the teeth of the mammal to be evaluated to obtain T-GLYC as a quantitative value (step of quantifying the total amount of glycated products contained in the evaluation target). (xix) Step of calculating PpG, which is a value of the weight ratio (amount of pyridinoline contained (ng / mg) / total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained)), from PYD obtained in the step of quantifying the amount of pyridinoline contained in the evaluation target (xvii) and T-GLYC obtained in the step of quantifying the total amount of glycated products contained in the evaluation target (xviii) (step of calculating the weight ratio PpG of the evaluation target). (xx) Step of quantifying the amount of pyridinoline contained (ng / mg) in collagen in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain R-PYD as a reference quantitative value (step of quantifying the amount of pyridinoline contained in the reference). (xxi) Quantifying the total glycated product content (ng / mg: converted to the content of pentosidine) of collagen contained in the teeth of the above-mentioned 1 or more mammals to obtain R-T-GLYC, which is a reference quantification value (reference total glycated product content quantification step), (xxii) Calculating R-PpG, which is a reference value of the weight ratio (content of pyridinoline (ng / mg) / total glycated product content (ng / mg: converted to the content of pentosidine)), from R-PYD obtained in the reference pyridinoline content quantification step (xx) and R-T-GLYC obtained in the reference total glycated product content quantification step (xxi) (reference weight ratio R-PpG calculation step), (xxiii) Comparing PpG calculated in the evaluation target weight ratio PpG calculation step (xix) with R-PpG calculated in the reference weight ratio R-PpG calculation step (xxii) (weight ratio PpG·R-PpG comparison step), As described above, it has the steps of (xvii) to (xxiii).

[0037] In the method using the quantification value of the pyridinoline content (ng / mg) of collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a disease in the mammal according to the present invention and / or whether the mammal is potentially healthy, the "evaluation target pyridinoline content quantification step (i)"; and in the method using the value of the weight ratio calculated from the quantification value of the pyridinoline content and the quantification value of the pentosidine content of collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a disease in the mammal according to the present invention and / or whether the mammal is potentially healthy, the "evaluation target pyridinoline content quantification step (x)"; and in the method using the value of the weight ratio calculated from the quantification value of the pyridinoline content and the quantification value of the total glycated product content of collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a disease in the mammal according to the present invention and / or whether the mammal is potentially healthy, the "evaluation target pyridinoline content quantification step (xvii)" are the same step.

[0038] Also, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, the "reference content pyridinoline amount quantification step (ii)" in the method using the quantitative value of the content pyridinoline amount (ng / mg) of collagen contained in the teeth of the mammal, and as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, the "reference content pyridinoline amount quantification step (xiii)" in the method using the value of the weight ratio calculated from the quantitative value of the content pyridinoline amount and the quantitative value of the content pentosidine amount of collagen contained in the teeth of the mammal, and as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, the "reference content pyridinoline amount quantification step (xx)" in the method using the value of the weight ratio calculated from the quantitative value of the content pyridinoline amount and the quantitative value of the total glycation product amount of collagen contained in the teeth of the mammal are the same step.

[0039] Also, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, the "evaluation target content pentosidine amount quantification step (iv)" in the method using the quantitative value of the content pentosidine amount (ng / mg) of collagen contained in the teeth of the mammal, and as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, the "evaluation target content pentosidine amount quantification step (xi)" in the method using the value of the weight ratio calculated from the quantitative value of the content pyridinoline amount and the quantitative value of the content pentosidine amount of collagen contained in the teeth of the mammal are the same step.

[0040] Also, in the method of using the quantitative value of the content of pentosidine (ng / mg) in the collagen contained in the teeth of a mammal as an index for evaluating the potential risk of developing a disease in the mammal according to the present invention and / or whether the mammal is potentially healthy, the "reference pentosidine content quantification step (v)" and, as an index for evaluating the potential risk of developing a disease in the mammal according to the present invention and / or whether the mammal is potentially healthy, the "reference pentosidine content quantification step (xiv)" in the method of using the value of the weight ratio calculated from the quantitative value of the content of pyridinoline and the quantitative value of the content of pentosidine in the collagen contained in the teeth of a mammal are the same step.

[0041] Also, in the method of using the quantitative value of the total glycated product content (ng / mg: converted to pentosidine content) in the collagen contained in the teeth of a mammal as an index for evaluating the potential risk of developing a disease in the mammal according to the present invention and / or whether the mammal is potentially healthy, the "evaluation target total glycated product content quantification step (vii)" and, as an index for evaluating the potential risk of developing a disease in the mammal according to the present invention and / or whether the mammal is potentially healthy, the "evaluation target total glycated product content quantification step (xviii)" in the method of using the value of the weight ratio calculated from the quantitative value of the content of pyridinoline and the quantitative value of the total glycated product content in the collagen contained in the teeth of a mammal are the same step.

[0042] In addition, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, the "reference total glycated product amount quantification step (viii)" in the method using the quantitative value of the total amount of glycated products contained in the collagen in the teeth of a mammal (ng / mg: converted to the amount of contained pentosidine), and as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, the "reference total glycated product amount quantification step (xxi)" in the method using the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in the collagen in the teeth of a mammal and the quantitative value of the total amount of glycated products are the same step.

[0043] Quantification of the amount of pyridinoline (ng / mg) in the "evaluated pyridinoline amount quantification step" and the "reference pyridinoline amount quantification step", quantification of the amount of pentosidine (ng / mg) in the "evaluated pentosidine amount quantification step" and the "reference pentosidine amount quantification step", and quantification of the total amount of glycated products (ng / mg: converted to the amount of contained pentosidine) in the "evaluated total glycated product amount quantification step" and the "reference total glycated product amount quantification step" can be performed using conventional preparation means known in the art. For example, it can be performed generally as follows.

[0044] (1) Weigh the dry collagen powder produced by the method for producing the collagen-rich organic composition according to the present invention into a test tube and swell it with a small amount of 6N hydrochloric acid under deaeration. (2) After adding 6N hydrochloric acid to the dry collagen and deaerating, seal it. (3) Perform hydrolysis by heat-treating at 110°C for 24 hours. (4) Cool the sample to room temperature. (5) Remove excess hydrochloric acid by drying under reduced pressure while heating to obtain a hydrolyzate of collagen. (6) Solubilize the obtained collagen hydrolyzate with an aqueous solution of 10% methanol. (7) Centrifuge at 12,000 rpm for 5 minutes at room temperature, and use the supernatant as the sample. (8) By adding a known amount of standard substance to the sample, the extraction rate and calibration curve can be determined, and the collagen content can be corrected as appropriate.

[0045] Also, the quantification method for the amount of pyridinoline contained (ng / mg) in the "Quantification step of the amount of pyridinoline contained in the evaluation target" and the "Quantification step of the amount of pyridinoline contained in the reference", the quantification method for the amount of pentosidine contained (ng / mg) in the "Quantification step of the amount of pentosidine contained in the evaluation target" and the "Quantification step of the amount of pentosidine contained in the reference", and the quantification method for the amount of total glycation products contained (ng / mg: converted amount of pentosidine contained) in the "Quantification step of the amount of total glycation products contained in the evaluation target" and the "Quantification step of the amount of total glycation products contained in the reference" are not particularly limited as long as their quantification is possible. As such a quantification method, for example, high performance liquid chromatography with fluorescence detector (HPLC-Flu) using heptafluorobutyric acid and formic acid as ion pair reagents can be employed.

[0046] The quantification method using high performance liquid chromatography with fluorescence detector (HPLC-Flu) using the heptafluorobutyric acid and formic acid as ion pair reagents can be carried out using conventional quantification means known in the art. For example, it can be carried out generally as follows.

[0047] (1) Using ultrapure water containing 0.05% heptafluorobutyric acid (mobile phase A) and a 1:1 mixture of methanol and ethanol containing 0.05% formic acid (mobile phase B) as the mobile phase, perform gradient elution and detection. (2) As specific elution conditions, start at a ratio of 90% mobile phase A and 10% mobile phase B at a flow rate of 0.5 mL / min at a detection column temperature of 40°C and elute for 0.5 minutes. (3) Then, increase the mobile phase B to 22% over 1 minute to create a concentration gradient, and then elute for 7.5 minutes as it is. (4) Subsequently, the mobile phase B was increased to 80% over 1 minute and then the liquid was pumped for another 1 minute. (5) Finally, after pumping with the mobile phase A at 10% and the mobile phase B at 90%, equilibration was carried out for 4 minutes at the ratio of the mobile phase A at 90% and the mobile phase B at 10%. (6) The above steps (1) to (5) are carried out for a total of 15 minutes per sample. A Cadenza CD-C18 column with a length of 150 mm and an inner diameter of 3 mm is used for the separation column, and a fluorescence detector is used for detection. In the first 8 minutes, it is monitored at an excitation wavelength of 295 nm and an emission wavelength of 395 nm, and in the last 7 minutes, it is monitored at an excitation wavelength of 325 nm and an emission wavelength of 385 nm. By these monitors, the contained pyridinoline is detected at around a retention time of about 6.5 minutes, and the contained pentosidine is detected at around a retention time of about 9.1 minutes.

[0048] Quantification of the amount of contained pyridinoline (ng / mg) in the "Quantification step of the amount of contained pyridinoline in the evaluation target" and the "Quantification step of the amount of contained pyridinoline in the reference", quantification of the amount of contained pentosidine (ng / mg) in the "Quantification step of the amount of contained pentosidine in the evaluation target" and the "Quantification step of the amount of contained pentosidine in the reference", and quantification of the amount of contained total glycation products (ng / mg: converted to the amount of contained pentosidine) in the "Quantification step of the amount of contained total glycation products in the evaluation target" and the "Quantification step of the amount of contained total glycation products in the reference" can be carried out for extracted teeth of mammals that are the subject of the evaluation of the potential risk of disease and / or whether the mammal is potentially healthy, mammals not suffering from the disease, mammals suffering from the disease, healthy mammals, and non-healthy mammals. In addition, it can also be carried out in modes such as cutting, contact, and non-contact.

[0049] Also, the mammals that are the subject of the evaluation of the potential risk of disease and / or whether the mammal is potentially healthy, mammals not suffering from the disease, mammals suffering from the disease, healthy mammals, and non-healthy mammals may all be the same individual.

[0050] In addition, as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the content of pyridinoline in collagen contained in the teeth of the mammal (ng / mg) may have other steps not only in the above-described "Quantification step (i) of the content of pyridinoline in the subject to be evaluated", "Quantification step (ii) of the content of pyridinoline in the reference", and "PYD·R-PYD comparison step (iii)", but also within a range that does not impair the features of the present invention.

[0051] In addition, as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the content of pentosidine in collagen contained in the teeth of the mammal (ng / mg) may have other steps not only in the above-described "Quantification step (iv) of the content of pentosidine in the subject to be evaluated", "Quantification step (v) of the content of pentosidine in the reference", and "PEN·R-PEN comparison step (vi)", but also within a range that does not impair the features of the present invention.

[0052] In addition, as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the total glycation product content in collagen contained in the teeth of the mammal (ng / mg: converted to the content of pentosidine) may have other steps not only in the above-described "Quantification step (vii) of the total glycation product content in the subject to be evaluated", "Quantification step (viii) of the total glycation product content in the reference", and "T-GLYC·R-T-GLYC comparison step (ix)", but also within a range that does not impair the features of the present invention.

[0053] In addition, as an index for evaluating the potential risk of developing a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of a weight ratio calculated from the quantitative value of the content of pyridinoline in collagen contained in the teeth of the mammal and the quantitative value of the content of pentosidine may include, in addition to the above-described "quantitative step (x) of the content of pyridinoline to be evaluated", "quantitative step (xi) of the content of pentosidine to be evaluated", "calculation step (xii) of the weight ratio PpP of the object to be evaluated", "quantitative step (xiii) of the content of pyridinoline in the reference", "quantitative step (xiv) of the content of pentosidine in the reference", "calculation step (xv) of the weight ratio R-PpP of the reference", and "comparison step (xvi) of the weight ratio PpP·R-PpP", and may also have other steps within a range that does not impair the features of the present invention.

[0054] In addition, as an index for evaluating the potential risk of developing a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of a weight ratio calculated from the quantitative value of the content of pyridinoline in collagen contained in the teeth of the mammal and the quantitative value of the content of total glycation products may include, in addition to the above-described "quantitative step (xvii) of the content of pyridinoline to be evaluated", "quantitative step (xviii) of the content of total glycation products to be evaluated", "calculation step (xix) of the weight ratio PpG of the object to be evaluated", "quantitative step (xx) of the content of pyridinoline in the reference", "quantitative step (xxi) of the content of total glycation products in the reference", "calculation step (xxii) of the weight ratio R-PpG of the reference", and "comparison step (xxiii) of the weight ratio PpG·R-PpG", and may also have other steps within a range that does not impair the features of the present invention.

[0055] As an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the amount of pyridinoline contained in the collagen in the teeth of the mammal (ng / mg), as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the amount of pentosidine contained in the collagen in the teeth of the mammal (ng / mg), as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a quantitative value of the total amount of glycated products contained in the collagen in the teeth of the mammal (ng / mg: converted amount of pentosidine contained), as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in the collagen in the teeth of the mammal and the quantitative value of the amount of pentosidine contained, and, as an index for evaluating the potential risk of a disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using a value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in the collagen in the teeth of the mammal and the quantitative value of the total amount of glycated products contained. In the method, the reason for limiting to "teeth" is that the pyridinoline crosslinking, pentosidine crosslinking, and crosslinking of total glycated products in type I collagen are all crosslinking reactions accelerated by factors such as in-vivo heat due to body temperature and sugars supplied transvascularly, and are crosslinking reactions caused according to the circulating blood flow rather than locally. Among them, it is considered that the collagen contained in teeth (especially dentin) most accurately reflects the accumulation of crosslinked molecules of pyridinoline crosslinked molecules, pentosidine crosslinked molecules, and total glycated products containing pentosidine crosslinked molecules. Teeth are the only tissue in the living body that is not incorporated into the metabolic cycle, and once formed, they are not absorbed and decomposed. Therefore, crosslinked molecules of pyridinoline crosslinked molecules, pentosidine crosslinked molecules, and total glycated products containing pentosidine crosslinked molecules accumulate in the teeth (especially dentin) containing collagen.

[0056] Furthermore, the collagen in teeth (especially dentin) is insoluble compared to bone. In pepsin digestion under the conditions of 0.01N hydrochloric acid, 4°C, and a treatment time of 72 hours, about 35% of the collagen in adult bovine bone is solubilized, while only 5.6% of the collagen in the dentin of adult bovine teeth is solubilized. Also, under the condition of pH 2, insoluble collagen such as skin and Achilles tendon swells to 4 - 8 times its volume, and the insoluble collagen in adult bovine bone swells to 1.2 times its volume, whereas the insoluble collagen in adult bovine dentin does not swell at all (edited by Hiroshi Nagai and Daisaburo Fujimoto, Collagen Experiment Methods, Kodansha Scientific, p.21 - 22). Also, the dentin collagen in teeth has the highest content rate of pyridinoline cross-linked molecules in the living body. Therefore, PYD (ng / mg), which is the quantitative value of the pyridinoline content in the collagen contained in mammalian teeth, PEN (ng / mg), which is the quantitative value of the pentosidine content in the collagen contained in mammalian teeth, T-GLYC (ng / mg: converted to pentosidine content), which is the quantitative value of the total glycated product content in the collagen contained in mammalian teeth, PpP (pyridinoline content (ng / mg) / pentosidine content (ng / mg)), which is the value of the weight ratio calculated from the quantitative value of the pyridinoline content (ng / mg) and the quantitative value of the pentosidine content (ng / mg) in the collagen contained in mammalian teeth, and PpG (pyridinoline content (ng / mg) / total glycated product content (ng / mg: converted to pentosidine content)), which is the value of the weight ratio calculated from the quantitative value of the pyridinoline content (ng / mg) and the quantitative value of the total glycated product content (ng / mg: converted to pentosidine content) in the collagen contained in mammalian teeth, it is reasonable to evaluate the potential risk of disease in mammals and / or whether the mammal is potentially healthy as an indicator.

[0057] On the other hand, the basis for excluding bone will be explained. As indicators of osteoporosis, numerical values of type I collagen C-terminal telopeptide (ICTP) in blood (serum) and NTx (type I collagen cross-linked N-telopeptide) in urine are widely used. However, these are substances that are released into the blood and urine as collagen degradation products due to matrix metalloproteinase (MMPs) activity and are substances containing pyridinoline cross-linked molecules. From this, in addition to the fact that it can be seen that the pyridinoline cross-linked molecules, pentosidine cross-linked molecules, and cross-linked molecules of total glycation products containing pentosidine cross-linked molecules contained in bone-containing collagen are partially cleared by metabolism, when using the excised bone, since bone is an organ containing a large amount of lipids due to yellow bone marrow, it is not easy to perform degreasing, and it becomes very difficult to purify highly pure collagen.

[0058] In addition, as an index for evaluating the potential risk of disease in mammals according to the present invention, a method using PYD, which is a quantitative value of the amount of pyridinoline contained in collagen in the teeth of mammals; as an index for evaluating the potential risk of disease in mammals according to the present invention, a method using PEN, which is a quantitative value of the amount of pentosidine contained in collagen in the teeth of mammals; as an index for evaluating the potential risk of disease in mammals according to the present invention, a method using T-GLYC, which is a quantitative value of the total amount of glycated products contained in collagen in the teeth of mammals; as an index for evaluating the potential risk of disease in mammals according to the present invention, a method using the value of PpP (amount of pyridinoline contained / amount of pentosidine contained), which is a weight ratio value calculated from PYD, which is a quantitative value of the amount of pyridinoline contained in collagen in the teeth of mammals, and PEN, which is a quantitative value of the amount of pentosidine contained; as an index for evaluating the potential risk of disease in mammals according to the present invention, a method using the value of PpG (amount of pyridinoline contained / total amount of glycated products contained), which is a weight ratio value calculated from PYD, which is a quantitative value of the amount of pyridinoline contained in collagen in the teeth of mammals, and T-GLYC, which is a quantitative value of the total amount of glycated products contained; as an index for evaluating whether a mammal according to the present invention is potentially healthy or not, a method using PYD, which is a quantitative value of the amount of pyridinoline contained in collagen in the teeth of mammals; as an index for evaluating whether a mammal according to the present invention is potentially healthy or not, a method using PEN, which is a quantitative value of the amount of pentosidine contained in collagen in the teeth of mammals; as an index for evaluating whether a mammal according to the present invention is potentially healthy or not, a method using T-GLYC, which is a quantitative value of the total amount of glycated products contained in collagen in the teeth of mammals; as an index for evaluating whether a mammal according to the present invention is potentially healthy or not, a method using the value of PpP (amount of pyridinoline contained / amount of pentosidine contained), which is a weight ratio value calculated from PYD, which is a quantitative value of the amount of pyridinoline contained in collagen in the teeth of mammals, and PEN, which is a quantitative value of the amount of pentosidine contained; and as an index for evaluating whether a mammal according to the present invention is potentially healthy or not,A method using the value of PpG (pyridinoline content / total glycated product content), which is a value of the weight ratio calculated from the quantitative value of PYD, which is the quantitative value of the pyridinoline content in the collagen contained in mammalian teeth, and the quantitative value of T-GLYC, which is the quantitative value of the total glycated product content, may be independent or related to each other.

[0059] Hereinafter, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the quantitative value of the pyridinoline content (ng / mg) of collagen contained in the teeth of a mammal, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the quantitative value of the pentosidine content (ng / mg) of collagen contained in the teeth of a mammal, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the quantitative value of the total glycated product content of collagen contained in the teeth of a mammal, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the value of the weight ratio (pyridinoline content (ng / mg) / pentosidine content (ng / mg)) calculated from the quantitative value of the pyridinoline content (ng / mg) and the quantitative value of the pentosidine content (ng / mg) of collagen contained in the teeth of a mammal, and, as an index for evaluating the potential risk of disease in a mammal according to the present invention and / or whether the mammal is potentially healthy, a method using the value of the weight ratio (pyridinoline content (ng / mg) / total glycated product content (ng / mg: pentosidine content conversion)) calculated from the quantitative value of the pyridinoline content and the quantitative value of the total glycated product content of collagen contained in the teeth of a mammal will be described based on examples. It should be noted that the technical scope of the present invention is not limited to the embodiments shown by these examples.

Examples

[0060] Example 1: Measurement of the Amounts of Pyridinoline, Pentosidine, and Total Glycation Products in Collagen Derived from Teeth (Primarily Dentin) of Human Non-Diabetic Patients, Human Diabetic Patients, and Cows The amounts of pyridinoline (ng / mg), pentosidine (ng / mg), and total glycation products (ng / mg: converted to the amount of contained pentosidine) in collagen derived from teeth (primarily dentin) of 14 human non-diabetic patients, 4 human diabetic patients, and 19 cows (cow specified risk materials; SRM) were measured according to the following procedure.

[0061] 1. Preparation of Collagen Powder [1-1] The hard tissue was ground to a particle size of about 1 mm using a grinder. [1-2] The attached soft tissue was removed by stirring in a 1% aqueous sodium carbonate solution at 70 °C for several hours. [1-3] Demineralization treatment was performed. Specifically, using a demineralization solution containing 1 M phosphoric acid and ethanol, demineralization treatment was carried out for several hours while reducing the pressure, and then replaced with the demineralization solution diluted several times, and demineralization treatment was carried out for several hours while reducing the pressure. Subsequently, using the above demineralization solution diluted 5 times, shaking was performed at room temperature for about one day and night to completely demineralize. Thereafter, washing was performed several times at room temperature using 5% ethanol. [1-4] The demineralized hard tissue was transferred to a mortar and pulverized into fine powder by rubbing in 70% ethanol. [1-5] Stirring was performed with 100% ethanol for washing, degreasing, and dehydration. [1-6] The ethanol remaining in the collagen was completely removed by drying to obtain dry collagen powder.

[0062] 2. Preparation of Samples for Analysis The dry collagen powder obtained in [2-1][1-6] was hydrolyzed. Specifically, the obtained dry collagen powder was weighed into a test tube and sufficiently swollen with a small amount of 6N hydrochloric acid under deaeration. Then, an appropriate amount of 6N hydrochloric acid was added to the dry collagen, and the test tube was welded and sealed under deaeration. The sealed test tube was hydrolyzed by heat treatment overnight at 110 °C for one day and night, then cooled to room temperature, the test tube was cut, and excess hydrochloric acid was removed by drying under reduced pressure while heating to obtain a hydrolyzate of collagen. [2-2] The obtained hydrolyzate of collagen was solubilized with a 10% aqueous methanol solution, centrifuged at 12,000 rpm for 5 minutes at room temperature, and the supernatant was used as a sample. [2-3] By adding a known amount of standard substance to the sample, the extraction rate and calibration curve were determined, and the collagen content was corrected as appropriate.

[0063] 3. Quantification of the amount of pyridinoline contained, the amount of pentosidine contained, and the amount of total glycation products contained, and calculation of their weight ratios by high-performance liquid chromatography with fluorescence detector (HPLC-Flu) using heptafluorobutyric acid and formic acid as ion-pair reagents Using ultrapure water containing 0.05% heptafluorobutyric acid (mobile phase A) and a 1:1 methanol-ethanol mixture containing 0.05% formic acid (mobile phase B) as the mobile phase, the liquid was delivered and detected under gradient settings. Specifically, starting at a ratio of 90% mobile phase A and 10% mobile phase B, the liquid was delivered at a flow rate of 0.5 mL / min at a detection column temperature of 40°C for 0.5 minutes. Subsequently, over 1 minute, the mobile phase B was increased to 22% to create a concentration gradient, and then the liquid was delivered as it was for 7.5 minutes. Subsequently, over 1 minute, the mobile phase B was increased to 80%, and then the liquid was delivered for an additional 1 minute. Finally, after delivering the liquid at a ratio of 10% mobile phase A and 90% mobile phase B, the mobile phase B was then decreased to 10%, and equilibrium was achieved for 4 minutes at a ratio of 90% mobile phase A and 10% mobile phase B. Subsequently, each sample was detected over a total of 15 minutes. A Cadenza CD-C18 column with a length of 150 mm and an inner diameter of 3 mm was used as the separation column, and a fluorescence detector was used for detection. In the first 8 minutes, it was monitored at an excitation wavelength of 295 nm and an emission wavelength of 395 nm, and in the last 7 minutes, it was monitored at an excitation wavelength of 325 nm and an emission wavelength of 385 nm. By this monitoring, the amount of pyridinoline contained was detected at around a retention time of approximately 6.5 minutes, and the amount of pentosidine contained and the amount of total glycation products contained were detected at around a retention time of approximately 9.1 minutes. From the obtained amount of pyridinoline contained and the amount of total glycation products contained, their weight ratio (amount of pyridinoline contained (ng / mg) / amount of total glycation products contained (ng / mg: converted to amount of pentosidine)) was determined.

[0064] The distribution diagrams of PYD, which is the quantitative value of the amount of pyridinoline contained in collagen (ng / mg) in the teeth of 14 human non-diabetic patients, 4 human diabetic patients, and 19 cows, according to the tooth age, are shown in Figure 1. The distribution diagrams of PEN, which is the quantitative value of the amount of pentosidine contained in collagen (ng / mg) in the teeth of 14 human non-diabetic patients, 4 human diabetic patients, and 19 cows, according to the tooth age, are shown in Figure 2. The distribution diagrams of T-GLYC, which is the quantitative value of the total glycated product amount (ng / mg: converted to the amount of pentosidine contained) in collagen in the teeth of 14 human non-diabetic patients, 4 human diabetic patients, and 19 cows, according to the tooth age, are shown in Figure 3. The distribution diagrams of PpP, which is the value of the weight ratio calculated from PYD, which is the quantitative value of the amount of pyridinoline contained in collagen (ng / mg), and PEN, which is the quantitative value of the amount of pentosidine contained (ng / mg) in the teeth of 14 human non-diabetic patients, 4 human diabetic patients, and 19 cows, according to the tooth age, are shown in Figure 4. The distribution diagrams of PpG, which is the value of the weight ratio calculated from PYD, which is the quantitative value of the amount of pyridinoline contained in collagen (ng / mg), and T-GLYC, which is the quantitative value of the total glycated product amount (ng / mg: converted to the amount of pentosidine contained) in the teeth of 14 human non-diabetic patients, 4 human diabetic patients, and 19 cows, according to the tooth age, are shown in Figure 5. In Figure 1, the dashed line indicates the approximate curve (linear approximation) based on the PYD of 14 human non-diabetic patients, corresponding to R-PYD. In Figure 2, the dashed line indicates the approximate curve (linear approximation) based on the PEN of 14 human non-diabetic patients, corresponding to R-PEN. In Figure 3, the dashed line indicates the approximate curve (linear approximation) based on the T-GLYC of 14 human non-diabetic patients, corresponding to R-T-GLYC. In Figure 4, the dashed line indicates the approximate curve (linear approximation) based on the PpP of 14 human non-diabetic patients, corresponding to R-PpP. In Figure 5, the dashed line indicates the approximate curve (linear approximation) based on the PpG of 14 human non-diabetic patients, corresponding to R-PpG.

[0065] Regarding human teeth, permanent teeth are the subject of measurement. Considering the formation period of permanent dentin, it can be considered that dentin formation starts in the tooth germ in the jawbone on average from around 10 years old, and accordingly, the formation of glycation crosslinks also starts (Investigation and Research on the Eruption Periods of Deciduous and Permanent Teeth in Japanese Children II, Journal of the Japanese Society of Pediatric Dentistry, 57(3), 363 - 373, 2019, 363). On the other hand, regarding the teeth in the lower jaw of bovine SRM, the same can be said on average 18 months after birth. Therefore, for the specified teeth of humans and bovine SRM, Table 1 below is used for the conversion from age to month age.

[0066] [Table 1] TIFF2025097590000002.tif110165

[0067] Also, PYD, which is the quantitative value of the amount of pyridinoline contained (ng / mg) in the collagen contained in the teeth of 14 human non - diabetic patients, PEN, which is the quantitative value of the amount of pentosidine contained (ng / mg), T - GLYC, which is the quantitative value of the total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained), PpP, which is the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained (ng / mg) and the quantitative value of the amount of pentosidine contained (ng / mg) (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), and PpG, which is the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained (ng / mg) and the quantitative value of the total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained) (amount of pyridinoline contained (ng / mg) / total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained)), which were quantified by high - performance liquid chromatography with fluorescence detector (HPLC - Flu) using heptafluorobutyric acid and formic acid as ion - pair reagents, are shown in Table 2 below.

[0068] [Table 2] TIFF2025097590000003.tif106165

[0069] Next, the quantitative values of PYD, which is the amount of pyridinoline contained (ng / mg) in the collagen contained in the teeth of 4 human diabetic patients, PEN, which is the amount of pentosidine contained (ng / mg), T-GLYC, which is the amount of total glycated products contained (ng / mg: converted to the amount of pentosidine contained), PpP, which is the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained (ng / mg) and the quantitative value of the amount of pentosidine contained (ng / mg) (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), and PpG, which is the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained (ng / mg) and the quantitative value of the amount of total glycated products contained (ng / mg: converted to the amount of pentosidine contained) (amount of pyridinoline contained (ng / mg) / amount of total glycated products contained (ng / mg: converted to the amount of pentosidine contained)), which were quantified by high performance liquid chromatography with fluorescence detector (HPLC-Flu) using heptafluorobutyric acid and formic acid as ion pair reagents, are shown in Table 3 below.

[0070] [Table 3] TIFF2025097590000004.tif60165

[0071] Next, the quantitative values of PYD, which is the amount of pyridinoline contained (ng / mg) in the collagen contained in the teeth of 19 cows, PEN, which is the amount of pentosidine contained (ng / mg), T-GLYC, which is the amount of total glycated products contained (ng / mg: converted to the amount of pentosidine contained), PpP, which is the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained (ng / mg) and the quantitative value of the amount of pentosidine contained (ng / mg) (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), and PpG, which is the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained (ng / mg) and the quantitative value of the amount of total glycated products contained (ng / mg: converted to the amount of pentosidine contained) (amount of pyridinoline contained (ng / mg) / amount of total glycated products contained (ng / mg: converted to the amount of pentosidine contained)), which were quantified by high performance liquid chromatography with fluorescence detector (HPLC-Flu) using heptafluorobutyric acid and formic acid as ion pair reagents, are shown in Table 4 below.

[0072] [Table 4] TIFF2025097590000005.tif80165

[0073] As shown in Fig. 1, it was revealed that the amount of pyridinoline contained in the collagen in the teeth of human diabetic patients (ng / mg) was slightly increasing with the increase in the age in months, as compared with the amount of pyridinoline contained in the collagen in the teeth of human non-diabetic patients and cows.

[0074] Also, as shown in Fig. 2, it was revealed that the amount of pentosidine contained in the collagen in the teeth of human diabetic patients (ng / mg) was increasing with the increase in the age in months, as compared with the amount of pentosidine contained in the collagen in the teeth of human non-diabetic patients and cows.

[0075] Also, as shown in Fig. 3, it was revealed that the total amount of glycated products contained in the collagen in the teeth of human diabetic patients (ng / mg: converted to the amount of pentosidine contained) was increasing with the increase in the age in months, as compared with the total amount of glycated products contained in the collagen in the teeth of human non-diabetic patients and cows.

[0076] Also, as shown in Fig. 4, it was revealed that the value of PpP (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), which is a value of the weight ratio calculated from PYD which is the quantitative value of the amount of pyridinoline contained in the collagen in the teeth of human non-diabetic patients and cows (ng / mg) and PEN which is the quantitative value of the amount of pentosidine contained (ng / mg), was decreasing with the increase in the age in months, as compared with the value of PpP (amount of pyridinoline contained (ng / mg) / amount of pentosidine contained (ng / mg)), which is a value of the weight ratio calculated from PYD which is the quantitative value of the amount of pyridinoline contained in the collagen in the teeth of human diabetic patients (ng / mg) and PEN which is the quantitative value of the amount of pentosidine contained (ng / mg).

[0077] Furthermore, as shown in Fig. 5, compared with PpG, which is a value of weight ratio calculated from PYD, which is the quantitative value of the amount of pyridinoline contained in collagen (ng / mg) in the teeth of human non-diabetic patients and cows, and T-GLYC, which is the quantitative value of the total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained), it was revealed that PpG, which is a value of weight ratio calculated from PYD, which is the quantitative value of the amount of pyridinoline contained in collagen (ng / mg) in the teeth of human diabetic patients, and T-GLYC, which is the quantitative value of the total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained), decreases with age.

[0078] From the above, it is clear that these can be used as indicators for evaluating the potential risk of disease in mammals and / or whether a mammal is potentially healthy. Therefore, as an indicator for evaluating the potential risk of disease in mammals and / or whether a mammal is potentially healthy, a method using the quantitative value of the amount of pyridinoline contained in collagen in the teeth of mammals, such as the method according to the present invention, is effective. As an indicator for evaluating the potential risk of disease in mammals and / or whether a mammal is potentially healthy, a method using the quantitative value of the amount of pentosidine contained in collagen in the teeth of mammals, such as the method according to the present invention, is effective. As an indicator for evaluating the potential risk of disease in mammals and / or whether a mammal is potentially healthy, a method using the quantitative value of the total amount of glycated products contained in collagen in the teeth of mammals, such as the method according to the present invention, is effective. As an indicator for evaluating the potential risk of disease in mammals and / or whether a mammal is potentially healthy, a method using the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in collagen and the quantitative value of the amount of pentosidine contained in collagen in the teeth of mammals, such as the method according to the present invention, is effective. Also, as an indicator for evaluating the potential risk of disease in mammals and / or whether a mammal is potentially healthy, it has been shown that a method using the value of the weight ratio calculated from the quantitative value of the amount of pyridinoline contained in collagen and the quantitative value of the total amount of glycated products contained in collagen in the teeth of mammals, such as the method according to the present invention, is effective.

Claims

1. A method using a quantitative value of the amount of pyridinoline contained in collagen in the teeth of a mammal as an indicator for evaluating the potential risk of a disease in a mammal and / or whether a mammal is potentially healthy, comprising: quantifying the amount of pyridinoline contained in collagen in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, to obtain a quantitative value PYD; quantifying the amount of pyridinoline contained in collagen in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals, to obtain a reference quantitative value R-PYD; comparing the PYD and the R-PYD; and a method comprising the same.

2. A method using a quantitative value of the amount of pentosidine contained in collagen in the teeth of a mammal as an indicator for evaluating the potential risk of a disease in a mammal and / or whether a mammal is potentially healthy, comprising: quantifying the amount of pentosidine contained in collagen in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, to obtain a quantitative value PEN; quantifying the amount of pentosidine contained in collagen in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals, to obtain a reference quantitative value R-PEN; comparing the PEN and the R-PEN; and a method comprising the same.

3. A method using a quantitative value of the total amount of glycation products contained in collagen in the teeth of a mammal as an indicator for evaluating the potential risk of a disease in a mammal and / or whether a mammal is potentially healthy, comprising: quantifying the total amount of glycation products contained in collagen in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy, in terms of ng / mg of the amount of pentosidine contained, to obtain a quantitative value T-GLYC; Quantifying the total glycated product content (ng / mg: converted to the content of pentosidine) of collagen contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain R-T-GLYC, which is a reference quantitative value, and comparing the T-GLYC and the R-T-GLYC A method comprising the steps of:

4. A method using a value of a weight ratio calculated from a quantitative value of the content of pyridinoline and a quantitative value of the content of pentosidine of collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a mammal suffering from a disease and / or whether the mammal is potentially healthy, comprising: Quantifying the content of pyridinoline (ng / mg) of collagen contained in the teeth of a mammal to be evaluated for the potential risk of a disease and / or whether the mammal is potentially healthy to obtain PYD, which is a quantitative value, and Quantifying the content of pentosidine (ng / mg) of collagen contained in the teeth of the mammal to be evaluated to obtain PEN, which is a quantitative value, and Calculating PpP (content of pyridinoline (ng / mg) / content of pentosidine (ng / mg)), which is a value of a weight ratio, from the obtained PYD and the obtained PEN, and Quantifying the content of pyridinoline (ng / mg) of collagen contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals to obtain R-PYD, which is a reference quantitative value, and Quantifying the content of pentosidine (ng / mg) of collagen contained in the teeth of the one or more mammals to obtain R-PEN, which is a reference quantitative value, and Calculating R-PpP (content of pyridinoline (ng / mg) / content of pentosidine (ng / mg)), which is a reference value of a weight ratio, from the obtained R-PYD and the obtained R-PEN, and comparing the PpP and the R-PpP A method comprising the steps of:

5. A method using a value of a weight ratio calculated from a quantitative value of the content of pyridinoline and a quantitative value of the content of total glycated products of collagen contained in the teeth of a mammal as an index for evaluating the potential risk of a mammal suffering from a disease and / or whether the mammal is potentially healthy, comprising: Quantifying the amount of pyridinoline in collagen contained in the teeth of a mammal to be evaluated for the potential risk of disease and / or whether the mammal is potentially healthy (ng / mg) to obtain PYD as a quantitative value, Quantifying the total amount of glycated products in collagen contained in the teeth of the mammal to be evaluated (ng / mg: converted to the amount of pentosidine contained) to obtain T-GLYC as a quantitative value, Calculating PpG, which is a value of the weight ratio (amount of pyridinoline contained (ng / mg) / total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained)), from the obtained PYD and the obtained T-GLYC, Quantifying the amount of pyridinoline in collagen contained in the teeth of one or more mammals selected from mammals not suffering from a disease, mammals suffering from a disease, healthy mammals, and non-healthy mammals (ng / mg) to obtain R-PYD as a reference quantitative value, Quantifying the total amount of glycated products in collagen contained in the teeth of the one or more mammals (ng / mg: converted to the amount of pentosidine contained) to obtain R-T-GLYC as a reference quantitative value, Calculating R-PpG, which is a reference value of the weight ratio (amount of pyridinoline contained (ng / mg) / total amount of glycated products contained (ng / mg: converted to the amount of pentosidine contained)), from the obtained R-PYD and the obtained R-T-GLYC, Comparing the PpG and the R-PpG A method comprising the steps of.

6. The method according to claim 1, wherein the quantifying step is a step of quantifying using high performance liquid chromatography with a fluorescence detector (HPLC-Flu) using heptafluorobutyric acid and formic acid as ion pair reagents.

7. The method according to any one of claims 1 to 6, wherein the mammal is one or more mammals selected from the group consisting of humans, cows, horses, pigs, wild boars, sheep, deer, dogs, and cats.

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