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    • 专利摘要:
    Method of obtaining useful data for the screening and diagnosis of osteoporosis. The present invention relates to the use of the composition for the treatment of osteoporosis, the use of markers and a method of obtaining useful data for the screening, diagnosis, prognosis and/or monitoring of osteoporosis, kit or device and uses . (Machine-translation by Google Translate, not legally binding)
    公开号:ES2687220A1
    申请号:ES201700255
    申请日:2017-03-23
    公开日:2018-10-24
    发明作者:María José MARTÍNEZ RAMÍREZ；Cristina MONTES CASTILLO
    申请人:Servicio Andaluz de Salud；
    IPC主号:
    专利说明:

    METHOD OF OBTAINING USEFUL DATA FOR THE SCREENING AND DIAGNOSIS OF OSTEOPOROSIS
    FIELD OF THE INVENTION
    S 10 The present invention is within the field of biomedicine and biotechnology, and refers to a method for screening, diagnosis, prognosis and / or monitoring of osteoporosis for the treatment of those individuals selected through said method. Said screening, diagnosis, prognosis and / or monitoring is performed by quantifying a series of markers: glucagon-like peptide markers (GLP-1), glucagon-like peptide 2 (GLP-2), gastric inhibitor polypeptide (GIP), VV peptide , amylin, dipeptidyl peptidase 4 (DPP-4), 25-0H vitamin D, calcium, phosphorus, magnesium, parathyroid hormone (PTH), osteocalcin, aminoterminal propeptide propeptide, and beta-ctx collagen carboxyterminal telypeptide (CTX-Beta cross -Iaps).
    fifteen BACKGROUND OF THE INVENTION
    twenty The great progress made in recent years around the action that certain intestinal peptides play in relation to glucose metabolism and its application in the treatment of diabetes mellitus, has opened new avenues of research on the action of these substances in other areas, having shown the existence of a relationship of these intestinal hormones in bone health, especially in osteoporosis.
    25 Osteoporosis and its main complication, osteoporotic fractures, constitute a serious public health problem, being responsible for excess mortality, morbidity. chronic pain, greater degree of dependence, admission to institutions and economic costs seriously affecting the quality of life of people who could enjoy the last years of their lives (Baim et al 2008).
    30 Osteoporosis is defined as "Skeletal disorder characterized by impaired bone resistance that predisposes to an increased risk of fracture. Bone resistance is the reflection of the integration of two main features: bone density and qualityB • Tissue deterioration Bone that occurs in osteoporosis is the result of a decrease in bone mass and the alteration of bone microarchitecture, due to imbalance in the processes of bone formation and reabsorption (Baim et al 2008).
    Bone remodeling is a complex system of homeostasis of bone mass, with an efficient self-control system, regulated by numerous factors, including genetic, mechanical, hormonal, nutritional and growth factors. Under normal circumstances, the resorption and formation processes are closely coupled and the result is that there is no net change in bone mass (Baim et al
    2008).
    Bone remodeling occurs with a circadian rhythm, having demonstrated an elevation of bone remodeling markers during the night and their decrease during the day (Yavropoulou & Yovos 2013), with bone resorption phenomena prevailing at night (Henriksen et al. 2007). Bone formation markers also follow a circadian rhythm, but the magnitude of the variation is shorter than that of reabsorption. Bone formation decreases after 4 days of fasting, specifically, osteocalcin (bone formation parameter) does not change after the first 24 hours of fasting (Walsh & Henriksen, 2010). An attempt has been made to explain this fact by the parallelism in the circadian secretion of hormones such as cortisol, paratohormone (PTH) or melatonin, but so far, this assumption has not been confirmed (Yavrapaulou & Yovos 2013).
    The key aspect in this sequence of bone remodeling rhythm has been to verify how bone resorption is significantly affected during fasting periods (Walsh & Henriksen, 2010), having proposed that the main cause of the circadian rhythm of bone remodeling is variation in food intake. The rate of remodeling is affected depending on food intake, increasing at night with the night fast (Deal 2009). As previously mentioned, the most affected process is resorption, while bone formation parameters are not affected by nutrient intake, this fact being explained by the existence of a decoupling of the two processes at the postprandial moment ( Yavropoulou & Yavos 2013).
    The mechanisms by which food regulates bone turnover are complex, with several mediators involved. The intake of glucose, protein, fat and mixed intake is associated with a significant reduction in bone resorption markers, detectable twenty minutes after feeding (Henriksen et al 2003). Bone formation is also influenced, but it seems that its response to nutrients is less than reabsorption (Clowes et al 2002). The biochemical analysis of bone resorption demonstrates that food intake is the main cause of reduced bone resorption during the day, which is followed by a nighttime increase in it (Bjarnason et al
    2002).
    The determining fact of the dramatic reduction in bone resorption that occurs after the ingestion of a meal along with other significant tests such as that the response of bone remodeling to oral glucose administration is much greater than when this same glucose is administered via intravenously (Walsh & Henriksen, 2010), or the fact that during the digestive rest that occurs with parenteral nutrition there is a reduction in bone mass, reaffirms the idea of a functional relationship between intestine and bone metabolism, which could possibly be mediated by hormones, which would respond to nutrient absorption (Clowes et al 2005, Maibellau et al 2013).
    It has been shown that the most important effectors of the acute response that occurs in bone metabolism in relation to food intake are intestinal and pancreatic peptides (Walsh & Henriksen, 2010).
    Intestinal peptides are a set of peptides (some hormonal) that are produced in the digestive system and are involved in the regulation of the different stages of digestion. Unlike other hormones, these peptides are not usually produced by specific glands, but are secreted to the extracellular fluid and blood by isolated endocrine cells distributed along the digestive tract, constituting what some authors call the intrinsic endocrine system of the gastrointestinal system . On other occasions, they are produced by neurons of the mesenteric and submucosal plexus, or by pancreatic islets. Intestinal peptides are released in response to different stimuli present in the intestinal lumen, how pH, mechanical strain, tonicity and presence of nutrients (Martinez de Victoria 2010).
    The investigation of these peptides has been of fundamental interest in recent years especially in relation to diabetes mellitus, developing the concept of ncretinas, which are defined as intestinal peptides released after food ingestion and that have a modulating action on glucose homeostasis, leading to the design of new drugs for the treatment of diabetes mellitus, but has also revealed the implication of these substances in other areas of intermediate metabolism, highlighting their effects at the level of bone tissue. Thus, the existence of a relationship of these hormones with bone formation and resorption has been shown, with preliminary data suggesting positive effects of several of these intestinal peptides on the regulation of bone resorption that occurs in response to food. This regulation would be very complex and different mediators would participate in it.
    The most important intestinal peptides that have been related to bone remodeling are: glucagon-like peptide 1 and 2 (GLP -1, GLP -2), insulinotropic dependent peptide
    of glucose (GIP), also called gastro · inhibitor peptide, and to a lesser extent the Peptide
    VV, and hormones produced by pancreatic beta cells: such as amylin, and
    pancreatic polypeptide (PP) (Walsh & Henriksen 2010). Fundamental would be in addition to
    Contest of these peptides, substances such as peptide receptor analogues
    5 similar to glucagon · 1 (GLP-1 RA), such as those derived from exendin 4
    (exenatide) and the enzyme dipeptidyl peptidase 4 (DPPAi) inhibitors, enzyme
    responsible for the metabolization of some of these peptides such as GLP-1 and 2
    (Dicembrini et al 2012). The peptides that have been associated with the incretin effect are
    fundamentally the GLP-1 and the GIP, which have a hypoglycemic effect,
    10 controlling and even decreasing body weight, which has motivated the development of
    mainly GLp · 1 analogue drugs and DPP-4 inhibitors, which are currently
    used in the treatment of diabetes mellitus.
    Different studies have shown the relationship between the aforementioned peptides
    and bone remodeling and osteoporosis. In 2003, Henriksen and collaborators in a double
    fifteen study conducted in healthy volunteers, show, as the intake of different
    Nutrient types (protein, fat, glucose and fructose) reduce bone resorption
    causing a parallel secretion of GLP-1 and GLP-2. In the same study the injection
    intravenous GLP-2 at different doses in 60 postmenopausal women causes a
    reduction of bone resorption, showing no action on the formation parameters
    twenty bone (Henriksen et al 2003). The decrease in bone resorption by GLP-2 requires a
    intact gastrointestinal tract, where GLP-2 receptors that have been located in the
    mesenteric plexus, be intact.
    Subsequently, another study by Henriksen's group administering GLP-2 for 14 days at
    a group of healthy postmenopausal women, shows, on the one hand the safety of
    2S treatment, and on the other the significant decrease in bone resorption, without affecting
    apparently bone formation, as evidenced by stable osteocalcin levels during
    the treatment (Henriksen et al 2007).
    In a trial in 160 postmenopausal women who received treatment with GLP-2, in
    on day 120 after the injection, he showed an increase in bone density in the hip and a
    30 reduction of the nocturnal increase in the C · telopeptide concentration, marker of
    bone resorption, while osteocalcin was not modified, a formation marker
    bone (Deal 2009).
    In contrast to the evidence of GLp · 2 in relation to bone formation and resorption
    which comes mainly from human studies, the physiological role of GLP-1 on
    5
    bone has only been investigated primarily in rodents In a study conducted in rats
    Wistar undergoing hyperlipidic diets, the administration of GLP-1, and its analogue,
    receptor stimulant, exendin, improve on the one hand the lipid metabolism and the
    glucose and on the other increases the expression of genes that encode osteocalcin and
    5 osteoprotegerin, reversing bone loss (Nuche-Berenguer et al 2011). In
    this same line, a recent study shows how exendine administered to rats
    ovariectomized promotes formation and decreases bone resorption (Ma et al 2013).
    Regarding GIP, it has been shown that there are receptors for GIP in osteoblasts, and
    that the stimulation of these receptors favors the synthesis of type 1 collagen and increases
    10 alkaline phosphatase activity (8011ag et al 2001), and as demonstrated in this study
    performed in rats, the administration of GIP causes an increase in measured bone mass
    by densitometry, curiously without increasing the weight of rodents, arguing
    that the GIP would be like a sentinel hormone that in addition to this direct action on the
    bone, could act maximizing the effect of nutrients and the action of other hormones
    fifteen (amylin, insulin) on bone tissue (801la9 et al 2001). Subsequently it has also
    demonstrated in a study conducted in cell cultures, that the GIP also acts on
    osteoclasts inhibiting bone resorption, thus corroborating the fact of
    that the GIP would mediate the effect that food intake has by inhibiting the
    bone resorption (Zhong et al 2007).
    twenty On the other hand it has been possible to verify how calcium from food is
    deposited in bone tissue through the GIP contest (Tsukiyama et al 2006). These
    authors also show how GIP has an action on calcium homeostasis, and
    an antiapoptotic action in osteoblasts, no action having been proven
    About osteoclasts In a later study conducted by the Maibel1au group, in
    2S mice with GIP receptor deficiency (GIPR-Knockout), is shown as the
    absence of this receptor favors an increase in bone mass by increasing the
    formation, but with a poor quality bone tissue (Gaudin-Audrain et al, 2013).
    We must also highlight a study that shows how in patients with
    Alteration of bone mass there is a decrease in different intestinal peptides. This
    30 study is performed in patients diagnosed with anorexia nervosa, showing a
    decrease in GIP and amylin values in relation to mass decrease
    bone (Wojcik et al 2010), although this study is limited by the small size of
    sample.
    On the other hand, it is known that in diabetes mellitus there is an increased risk of osteoporotic fracture (Dicembrini et al 2011), and it is recognized that treatment with GLP-1 analogues leads to a decrease in weight and the possible positive action of these In bone homeostasis, it could be eclipsed by weight loss, which as is known can lead to a decrease in bone mass (Labouesse 2014). However, it has been possible to verify in a trial with humans how the treatment with exenatide (GLP-1 receptor agonist) does not modify the risk of fracture (Bunck et al 2011).
    Osteoporosis is a chronic disease, and currently it cannot be cured, therefore its treatment is supposed to be undefined. Currently authorized drugs are: Estrogens, estrogen receptor stimulants, bisphosphonates, Denosumab, Teriparatide, Strontium ranelate (only authorized in very serious osteoporosis).
    Due to the scarcity of therapeutic means, contraindications and adverse effects, it is necessary to propose alternative treatments to be modified depending on the patient and their characteristics. what is called sequential Uterapia-, as proposed by the Spanish Society of Osteoporosis and Mineral Metabolism Research Guide (SEIOMM 2014). It is therefore necessary to investigate new lines of treatment that may be a safe and effective alternative for the treatment of osteoporosis.
    BRIEF DESCRIPTION OF THE INVENTION
    A first aspect of the invention relates to the use of a composition, hereinafter composition of the invention, comprising glucagon-like peptide 1 (GLP1), GLP-1 analogs, GLP-1 receptor agonists, glucagon-like peptide 2
    (GLP-2), GLP-2 analogs, GLP-2 receptor agonists, DPP-4 inhibitors, or any of its pharmaceutically acceptable salts, esters, tautomers, solvates and hydrates, or any combination thereof, in the preparation of a medicine for the prevention, relief, improvement and / or treatment of osteoporosis.
    Alternatively it refers to the use of a composition, consisting of the glucagon-like peptide (GLP-1), GLP-1 analogs, GLP-1 receptor agonists, glucagon-like peptide 2 (GLP-2), GLP-2 analogues, GLP-2 receptor agonists, OPP-4 inhibitors, or any of its pharmaceutically acceptable salts, esters, tautomers, solvates and hydrates, or any combination thereof, in the preparation of a medicament for the prevention, relief, improvement and / or treatment of osteoporosis.
    In a preferred embodiment of this aspect of the invention, it further comprises GIP, YY peptide, amylin or any of its pharmaceutically acceptable salts, esters, tautomers, solvates and hydrates.
    In another preferred embodiment, it refers to the GLP-1 receptor analogs and agonists being selected from the list consisting of repaglinide, nateglinide, exenatide, pramlintide, liraglutide, lixisenatide, albiglutide, dulaglutide or any combination thereof.
    In another preferred embodiment, it means that the GLP-2 analog consists of teduglutide.
    In another preferred embodiment, it refers to OPP-4 inhibitors being selected from the list consisting of sitagliptin, vildagliptin, saxagliptin, alogliptin, linagliptin, gemigliptin, sitagliptin and simvastatin, or any combination thereof.
    In another preferred embodiment of this aspect of the invention, the composition further comprises another active ingredient, preferably an agent that affects bone structure and mineralization that is selected from the list consisting of bisphosphonates, bone morphogenic proteins, strontium ranelate, denosumab, estrogens, estrogen receptor stimulants, raloxifene, teriparatide, parathyroid hormone, calcitonin, calcium, cholecalciferol, calcifediol or any combination thereof.
    In another preferred embodiment, the composition further comprises one or more pharmaceutically acceptable excipients.
    A second aspect of the invention relates to the use of glucagon-like peptide markers (GLP-1), carboxyterminal telopeptide of Beta-ctx collagen (CTX-Beta crosslaps), or their combination for screening, diagnosis, prognosis and / or monitoring osteoporosis.
    In a preferred embodiment of this aspect of the invention, it further comprises the glucagon-like peptide 2 (GLP-2), dipeptidyl peptidase 4 (OPP-4), or any combination thereof.
    In another preferred embodiment it further comprises the gastric inhibitor polypeptide (GI P), VV peptide, amylin, or any combination thereof.
    In another preferred embodiment it further comprises 25-0H vitamin O, calcium, phosphorus, magnesium, parathyroid hormone (PTH), or any combination thereof.
    In another preferred embodiment, it further comprises osteocalcin, pro-collagen aminoterminal propeptide, beta-ctx collagen carboxyterminal telopeptide (CTX-Beta cross-Iaps), or any combination thereof.
    A third aspect of the invention relates to a method of obtaining useful data for the screening, diagnosis, prognosis and / or monitoring of osteoporosis, hereinafter the first method of the invention, comprising:
    a) quantify the expression products GLP-1, GLP-2, OPP-4 and CTX-Beta cross-Iaps, as defined in the present invention, in an isolated biological sample from an individual who has been previously administered a nutritional preparation, and the
    10 Zscorelumbar, and
    b) calculate the probability of an individual suffering from osteoporosis based on the formula
    (one):
    Probability =, + e (-O.06B 'GI.PI.o.m' crx'.oI.411 'ZSCORELUMBAR.9.0L6)
    Formula 1)
    15 where
    GLP1: is the peptide value similar to glucagon 1 (GLP-1);
    CTX: is the value of the CTX Beta-cross laps bone remodeling parameter; Y
    Zscorelumbar: is the value of the lumbar Z score obtained by bone Densitometry (DEXA).
    In another preferred embodiment, the biological sample isolated in step a) is blood.
    A fourth aspect of the invention relates to a method for screening, diagnosis, prognosis and / or monitoring of osteoporosis, comprising steps (a) and (b) as described in the present invention, hereafter referred to as the second method. of the invention, and
    25 also includes:
    c) assign the individual to the group of individuals who have osteoporosis when the result of Formula I is greater than 0.5.
    In a preferred embodiment, it refers to the second method of the invention for the screening, diagnosis, prognosis and / or monitoring of osteoporosis.
    In the preferred embodiment, the biological sample isolated in step a) is blood.
    S A fifth aspect of the invention relates to the use of the composition of the invention for the prevention, relief, improvement and / or treatment of individuals selected by the methods of the invention.
    A sixth aspect of the invention relates to a kit or device, hereinafter kit or device of the invention, comprising a:
    component A, formed by a blood collector, and / or
    10 component B, consisting of one or more blood sample tubes with 1 O ~ L of an OPP · 4 inhibitor per mL of blood, a 96-well plate, conjugated magnetic beads, specific antibodies, standards, buffers, solvents , streptavidin · PE, filter plate, flat bottom plate, sealing tape, and / or
    fifteen component e, formed one or more blood sample tubes, tri potassium EDTA, sodium heparin, and so
    component D, formed by a light spectrum measuring device, and / or
    component E, formed by a densitometer, and / or
    or any of its combinations.
    twenty A seventh aspect of the invention relates to the use of the kit or device of the invention, for obtaining useful data for the screening, diagnosis, prognosis and / or monitoring of osteoporosis.
    An eighth aspect of the invention relates to a computer program comprising program instructions for making a computer carry out the process according to any of the methods of the invention.
    2S A ninth aspect of the invention relates to a computer-readable storage medium comprising program instructions capable of causing a computer to carry out the steps of any of the methods of the invention.
    A tenth aspect of the invention relates to a transmissible signal comprising program instructions capable of having a computer perform the steps of any of the methods of the invention.
    S DESCRIPTION OF THE FIGURES
    Figure 1. ROC curve with the values of the predicted probabilities of the model and the original dependent variable.
    Detailed description of the invention
    10 The authors of the present invention describe the use of a method for screening, diagnosis, prognosis and monitoring of osteoporosis by quantifying a series of markers, namely GLP-1, GLP-2, GIP, OPP-4 and CTX-Beta cross-Iaps .
    lS In this work an analysis was applied based on a case-control study, to look for potential markers, mainly the intestinal peptides GLP-1, GLP-2 and GIP, as well as the enzyme responsible for their metabolization, OPP-4, and CTX -Beta cross-Iaps that can help understand the pathological processes related to osteoporosis.
    twenty Blood samples were taken as biological material and analyzed by luminescence techniques. The set of significant markers identified, together with the lumbar Z-score were used to apply a multivariate logistic regression model to group patients in risk groups for osteoporosis for early detection or screening and diagnosis purposes, as well as with prevention, relief, improvement and treatment purposes.
    COMPOSITION OF THE INVENTION
    25 30 Therefore, a first aspect of the invention relates to the use of a composition, hereafter referred to as the invention, comprising glucagon-like peptide 1 (GLP-1), GLP-1 analogs, receptor agonists of GLP-1, glucagon-like peptide 2 (GLP-2), GLP-2 analogs, GLP-2 receptor agonists, DPP-4 inhibitors, or any of its salts, esters, tautomers, solvates and hydrates pharmaceutically acceptable, or any combination thereof, in the preparation of a medicament for the prevention, relief, improvement and treatment of osteoporosis.
    eleven
    Alternatively it refers to the use of a composition, which consists of the peptide similar to glucagon 1 (GLP-1 l, analogs of GLP-1, agonists of GLP-1 receptors, peptide similar to glucagon 2 (GLP-2), GLP-2 analogues, GLP-2 receptor agonists, OPP-4 inhibitors, or any of its pharmaceutically acceptable salts, esters, tautomers, soJvates and hydrates, or any combination thereof, in the preparation of a medicament for the prevention, relief, improvement and / or treatment of osteoporosis.
    In a preferred embodiment of this aspect of the invention, it further comprises GIP, YY peptide, amylin or any of its pharmaceutically acceptable salts, esters, tautomers, solvates and hydrates.
    In another preferred embodiment, it refers to the GLP-1 receptor analogs and agonists being selected from the list consisting of repaglinide, nateglinide, exenatide, pramlintide, liraglutide, lixisenatide, albiglutide, dulaglutide or any combination thereof.
    In another preferred embodiment, it means that the GLP-2 analog consists of teduglutide.
    In another preferred embodiment, it refers to OPP-4 inhibitors being selected from the list consisting of sitagliptin, vildagliptin, saxagliptin, alogliptin, linagliptin, gemigliptin, sitagliptin and simvastatin, or any combination thereof.
    In another preferred embodiment of this aspect of the invention, the composition further comprises another active ingredient, preferably an agent that affects bone structure and mineralization that is selected from the list consisting of bisphosphonates, bone morphogenic proteins, strontium ranelate, denosumab, estrogens, estrogen receptor stimulants, raloxifene, teriparatide, parathyroid hormone, calcitonin, calcium, cholecalciferol, calcifediol or any combination thereof.
    In another preferred embodiment, the composition further comprises one or more pharmaceutically acceptable excipients.
    In another preferred embodiment, administration is performed orally, sublingually, inhalatively, transdermally, intramuscularly (1M), intravenously (IV), or subcutaneously (Se).
    In another preferred embodiment of this aspect of the invention, any of the active ingredients of the present invention can be administered to an individual simultaneously, in combination, or sequentially, that is, that the active ingredients do not have to be
    interacting chemically as part of a composition, being juxtaposed without being part of a true combination. if notthatthey can
    S The term "prevention" as understood in the present invention consists in preventing the onset of the disease, that is, preventing the disease or pathological condition from occurring in a subject (preferably mammal, and more preferably a human), in particularly, when said subject has a predisposition for the pathological condition.
    10 The term "treatment" as understood in the present invention refers to combating the effects caused as a result of a disease or pathological condition of interest in a subject (preferably mammal, and more preferably a human) that includes:
    (i) inhibit the disease or pathological condition, that is, stop its development;
    (ii) alleviate the disease or the pathological condition, that is, cause the regression of the disease or the pathological condition or its symptomatology;
    (iH) stabilize the disease or pathological condition.
    15 20 Osteoporosis is understood as defined in the present invention as a skeletal disorder characterized by altered bone strength that predisposes to an increase in fracture risk. Bone resistance is the reflection of the integration of two main features: bone density and quality. The deterioration of bone tissue that occurs in osteoporosis is the result of a decrease in bone mass and the alteration of bone microarchitecture, due to imbalance in the processes of bone formation and reabsorption.
    2S Depending on its etiology, osteoporosis can be classified as primary or secondary. Primary osteoporosis is the most common type of osteoporosis. This diagnosis is established when, after evaluating the patient, the cause that causes it is not found. In turn, primary osteoporosis can be categorized as juvenile, postmenopausal, age-related and idiopathic.
    30 Secondary osteoporosis is diagnosed when bone loss is caused by another disease or by the use of particular drugs. Fractures Most often occur in the bones of the hip, the vertebrae of the spine and the bones of the wrist. Vertebral fractures can cause loss of height and deformity of the rib cage.
    13
    Osteoporosis does not cause symptoms and usually goes unnoticed, which is why it has been called "the silent epidemic." The clinical manifestations of osteoporosis appear as a consequence of its complications: fractures. Also, osteoporosis is a chronic disease, and currently has no cure, therefore its treatment is chronic. The first step before recommending a treatment is to evaluate the patient to determine if it is a primary or secondary osteoporosis, in order to detect the diseases that cause it, some of which usually go unnoticed. If the causative disease is treated appropriately and low bone density for age persists, the treatment will depend on the dynamics of the bones. The general guidelines are based on recommending an adequate amount of calcium in the diet, the practice of physical exercise and the use of medications that contribute to the maintenance or increase of bone mass. Currently authorized drugs are: estrogens, estrogen receptor stimulants, bisphosphonates, denosumab, teriparatide, strontium ranelate (only authorized in very serious osteoporosis).
    The term "active substance", KfármacoK "active substance", "pharmaceutically active substance
    ,
    active "," active ingredient "or" pharmaceutically active ingredient "as used herein means any component that potentially provides a pharmacological activity or other different effect in the diagnosis, cure, mitigation, treatment, or prevention of a disease, or that affects the structure or function of the body of man or other animals.The term includes those components that promote a chemical change in the preparation of the drug and are present therein in a planned modified form that provides the specific activity or effect .
    The term · glucagon-like peptide 1 GLP-1 "refers to a hormone derived from the transcription of a gene called proglucagon whose physiological function is based on blood glucose concentration. The main source of GLP-1 in the body is cells L of the intestine, which secrete the hormone as an intestinal product.The biologically active form of the hormone GLP-1 are GLP-1- (7-37) and GLP-1- (7-36) NH2.
    The term "GLP-1 analogues and GLP_1K receptor agonists refers to those active ingredients that have an agonist action of the GLP-1 receptor, also called incretin mimetics, are a type of drug with a ncretin action. These drugs, repaglinide, naleglinide, exenalide, pramlinlide, liraglulide, lixisenalide, albiglulide, and dulaglutide, copy or mimic the action of GLP-1 produced by the body.The effects of GLP ~ 1 last only a few minutes, but analogues and agonists of the GLP-1 receptors last approximately 10 hours.The GLP-1 receptor agonists are formulated in liquid form, which is injected under the skin of the stomach, thigh or arm.According to the drug of choice, one or twice a day, or once a week.
    The term ~ glucagon 2 GLP-2 "peptide refers to a 33 amino acid peptide with the sequence HADGSFSDEMNTILDNLAARDFINWLlQTKITD in humans. GLP-2 is produced by the intestinal endocrine L cell and by several neurons in the central nervous system. Intestinal GLP-2 is co-secreted together with GLP-1 after nutrient intake.When administered externally, GLP-2 produces a number of effects on humans and rodents, including intestinal growth, increased intestinal function. , the reduction of bone breakdown and neuroprotection GLP-2 can act in an endocrine way to link intestinal growth and metabolism with nutrient intake GLP-2 and related analogues can be treatments for bowel syndrome short, Crohn's disease, osteoporosis and as adjuvant therapy during cancer chemotherapy.
    The term "GLP-2 analogue" refers to those active ingredients that have an action similar to GLP-2, specifically teduglutide.
    The term ~ GIP gastric inhibitor polypeptide or glucosau-dependent insulinotropic peptide (from the English gastric inhibitory pofypeptide), is a member of the secretin hormone family. GIP, together with the glucagon-like peptide type 1 (GLP-1), belong to a class of molecules known as incretins. The function of the GIP is to induce insulin secretion, which is stimulated mainly by the hyperosmolarity of glucose in the duodenum. The amount of insulin secreted is greater when glucose is administered orally than intravenously.
    This specification is understood as ~ dipeptidjl peptidase-4 DPP-4, adenosine deaminase protein complexing 2 or CD26u as a protein that, in humans, is encoded by the DPP4 gene. OPP-4 exhibits enzymatic activity and acts by degrading the incremental hormones GLP-, and GIP. Inhibitors of this protein have been developed, the DPP-4 inhibitors that are being used as a new class of medications for the treatment of type 2 diabetes, specifically in clinical practice sitagliptin, vildagliptin, saxagliptin, alogliptin are being used. and linagliptin: These compounds block the degradation of incretinic hormones, such as GLP-1, and can increase their natural levels in circulation, favoring glycemic control in type 2 diabetic patients.
    When the "agents that affect bone structure and mineralization" are mentioned herein, it refers to all those active ingredients whose main action is based on treating skeletal disorders characterized by impaired bone resistance that predisposes to an increase of the risk of fracture. Preferably, these agents can treat bone density and quality.
    Here, when reference is made to the term "salts, esters, tautomers, solvates and hydrates" refers to any pharmaceutically acceptable salt, ester, tautomer, solvate or hydrate, or any other compound that, in its administration, is capable of provide (directly or indirectly) a compound such as those described herein. However, it will be noted that pharmaceutically unacceptable salts also fall within the scope of the invention, since these may be useful for the preparation of pharmaceutically acceptable salts. The preparation of salts, prodrugs and derivatives can be carried out by methods known in the state of the art.
    The term "solvate," as used herein, includes both pharmaceutically acceptable solvates, that is, solvates of the compounds present in the composition of the invention that can be used in the manufacture of a medicament, such as pharmaceutically acceptable solvates, which may be useful in the preparation of pharmaceutically acceptable solvates or salts.The nature of the pharmaceutically acceptable solvate is not critical as long as it is pharmaceutically acceptable.In a particular embodiment, the solvate is a hydrate.The solvates can be obtained by conventional solvation methods known by experts in the field.
    The term "therapeutically effective amount" refers to the amount of the agent or compound capable of developing the therapeutic action determined by its pharmacological properties, calculated to produce the desired effect and, in general, will be determined, among other causes, by the proper characteristics. of the compounds, in addition to the age, condition of the patient, the severity of the alteration or disorder, and the route and frequency of administration.
    In accordance with the EMEA definition, any component in the composition other than an active ingredient is considered excipient. An example of additional pharmaceutically acceptable excipients could be, but not limited to, those selected from the list consisting of: methanol, propylene glycol, ethylene glycol, dimethylsulfoxide, 2,3 butanediol, glycerol, and other alcohols, galactose, glucose,
    sucrose, trehalose and other carbohydrates or carbohydrates, polyvinylpyrrolidone, polyvinyl alcohol, sodium hyaluronidate and other polymers, or any combination thereof.
    5 The pharmaceutically acceptable adjuvants and vehicles that can be used in said compositions are the adjuvants and vehicles known to those skilled in the art and commonly used in the elaboration of therapeutic compositions.
    10 15 20 The pharmaceutical composition as mentioned in the invention can be administered topically, transdermally, orally, nasally, intramuscularly, intravenously, intraperitoneally, subcutaneously, enterally or parenterally. Illustrative examples of topical or transdermal administration include, but are not limited to, iontophoresis, sonophoresis, electroporation, mechanical pressure, osmotic pressure gradient, occlusive cure, microinjections, needleless injections by pressure, microelectric patches and any combination thereof. Illustrative examples of pharmaceutical forms of oral administration include tablets, capsules, granules, solutions, suspensions, etc., and may contain conventional excipients, such as binders, diluents, disintegrants, lubricants, humectants, etc., and may be prepared. by conventional methods. The pharmaceutical compositions can also be adapted for parenteral administration, in the form of, for example, sterile lyophilized solutions, suspensions or products, in the appropriate dosage form; in this case, said pharmaceutical compositions will include suitable excipients, tampons, surfactants, etc. In any case, the excipients will be chosen based on the pharmaceutical form of administration selected. A review of the different pharmaceutical forms of
    The administration of farm vest and its preparation can be found in the book MTratado de Farmacia Galénjca ~, by C. Fauli i Trillo, 10 Edition, 1993, Luzan 5, S.A. of Editions.
    2S The term "medication", as used herein, refers to any substance used for prevention, diagnosis, relieves, treatment or cure of diseases in man and animals.
    30 The compositions of the present invention can be used together with other medicaments in combination therapies. The other drugs may be part of the same composition or of a different composition, for administration at the same time or at different times.
    Both the compositions of the present invention, as well as the combined preparation can be formulated for administration to an animal, and more preferably to a mammal, including man, in a variety of ways known in the state of
    technique. Such combined compositions or preparations and / or their formulations may be administered to an animal, including a mammal and, therefore, to man, in a variety of ways, including, but not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intrathecal, intraventricular, oral, enteral, parenteral, intranasal or dermal.
    s Administration of the compositions or pharmaceutical forms of the present invention can be accomplished by any suitable method, such as intramuscular, subcutaneous, or intravenous infusion. Intramuscular administration is preferred for pharmacokinetic convenience and for the severity of the disease to be treated.
    10 The amount of active ingredients present in the composition of the invention will depend on the relative efficacy of the compounds chosen, the severity of the disease to be treated and the weight of the patient. It is important to keep in mind that it may be necessary to introduce variations in the dose, depending on the age and condition of the patient, as well as modifications in the route of administration.
    fifteen It should be emphasized that the term "combined preparation" or also called "juxtaposition", in this specification, means that the components of the combined preparation need not be present as a joint, for example in a composition, in order to be available for application separated or sequential. In this way, the expression Myuxtapuesta "implies that it is not necessarily a true combination, in view of the physical separation of the components.
    twenty
    USE OF BIOMARKERS
    2S A second aspect of the invention relates to the use of glucagon-like peptide markers (GLP-1), beta-ctx collagen carboxyterminal telopeptide (CTX-Beta crosslaps), or their combination for screening, diagnosis, prognosis and / or monitoring of osteoporosis.
    In a preferred embodiment of this aspect of the invention, it further comprises the glucagon-like peptide 2 (GLP-2), dipeptidyl peptidase 4 (DPP-4), or any combination thereof.
    30 In another preferred embodiment, it further comprises the gastric inhibitor pOpeptide (GIP), VY peptide, amylin, or any combination thereof.
    In another preferred embodiment it further comprises 25 · 0H vitamin D, calcium, phosphorus, magnesium, parathyroid hormone (PTH), or any combination thereof.
    In another preferred embodiment it further comprises osteocalcin, aminoterminal procollagen propeptide, carboxyterminal telopeptide of the Beta · ctx cOlagen (CTX-Beta cross-Iaps), or any combination thereof.
    In another preferred embodiment the markers are selected from the list consisting of GLP · 1, GLP-2, GIP, DPP-4, CTX-Beta cross · laps or any combination thereof. In a more preferred embodiment, the marker is GLP-1 and GLP-2. In an even more preferred embodiment, the marker is GLp · 1.
    The term ~ Marker, biomarker or biological marker · is used to designate that substance used as an indicator of a biological state. It should be able to be measured objectively and evaluated as an indicator of a normal biological process, pathogenic state or response to a pharmacological treatment.
    The term carboxyterminal Utelopeptide of the collagen Beta · ctx or CTX · Beta cross · laps ~ refers to an indicator of bone metabolism, and is an aid when assessing treatment with antiresortive drugs (bisphosphonates, denosumab, strontium ranelate) or formators (teriparatide, selective estrogen receptor stimulants).
    The term population screening or Screening "as used in the present invention refers to the strategy applied to a population to detect a disease in individuals without signs or symptoms of that disease early within a community. So that this strategy of Prevention be considered as screening must meet the criteria of Frame and Carlson:
    • The disease sought is a common cause of morbidity and mortality (prevalent disease).
    • Be detectable in presymptomatic stage.
    • Diagnostic tests must be effective and effective (acceptable sensitivity and specificity).
    • Early treatment should be better than in the symptomatic stage.
    • The potential damage of the intervention must be less than in non-early treatment.
    The term ~ diagnosis · in this report refers to a procedure by which a disease, nosological entity, syndrome, or any state of health or illness is identified.
    In the present invention, "prognosis" is understood as the expected evolution of a disease and refers to the assessment of the probability according to which a subject suffers from a disease as well as the assessment of its onset, stage of development, evolution, or its regression, and / or the prognosis of the course of the disease in the future. As those skilled in the art will understand, such assessment, although preferred, may not be correct for 100% of the subjects to be diagnosed. The term, however, requires that a statistically significant part of the subjects can be identified as having the disease or having a predisposition to it. If a part is statistically significant, it can be determined by the person skilled in the art using several well-known statistical evaluation tools, for example, determination of confidence intervals, determination of p-values, Student's t-test, Mann's test -Whitney, etc. Preferred confidence intervals are at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%. P values are preferably 0.2, 0.1, 0.05. In turn, according to the method of the present invention, other subclassifications could be established within this principal, thus facilitating the choice and establishment of appropriate therapeutic regimens or treatment. This discrimination, as understood by one skilled in the art, is not intended to be correct in 100% of the samples analyzed. However, it requires that a statistically significant amount of the analyzed samples be classified correctly. The amount that is statistically significant can be established by a person skilled in the art by using different statistical tools, for example, but not limited, by determining confidence intervals, determining the significance value P, Student test or discriminant functions. Fisher, non-parametric measurements of Mann Whitney, Spearman correlation, logistic regression, linear regression, area under the ROC curve (AUC). Preferably, the confidence intervals are at least 90%, at least 95%, at least 97%, at least 98% or at least 99%. Preferably, the value of p is less than 0.1, 0.05, 0.01, 0.005 or 0.0001. Preferably, the present invention makes it possible to correctly detect the disease differentially by at least 60%, more preferably at least 70%, much more preferably at least 80%, or even much more preferably at least 90%. of the subjects of a certain group or population analyzed.
    The term useguimiento · as it is reflected in this report refers to a health care process that continues to another diagnostic or therapeutic intervention, with the aim of ending the episode of care initiated to achieve its full recovery, or to maintain a state of Satisfactory health in chronic diseases.
    Mtc ALL OF THE INVENTION
    A third aspect of the invention relates to a method of obtaining useful data for the screening, diagnosis, prognosis and / or monitoring of osteoporosis, hereinafter the first method of the invention, comprising:
    a) quantify the expression products GLP-1, GLP-2, OPP-4 and CTX-Beta cross-Iaps, as defined in the present invention, in an isolated biological sample from an individual who has been previously administered a nutritional preparation, and the
    10 Zscorelumbar, and
    b) calculate the probability of an individual suffering from osteoporosis based on formula (1):
    . . one
    Probability = (-0068 'GLPI-om' crX-4418 'ZSCORELUMBAR.9016)
    l + e '...
    Formula 1)
    15 where
    GLP1: is the peptide value similar to glucagon 1 (GLP-1);
    CTX: is the value of the CTX Beta-cross laps bone remodeling parameter; Y
    Zscorelumbar: is the value of the lumbar Z score obtained by bone Densitometry
    (OEXA).
    In a preferred embodiment, it refers to the first method of the invention for screening, diagnosis, prognosis and / or monitoring of osteoporosis.
    In another preferred embodiment, the biological sample isolated in step a) is blood. Preferably the blood is extracted from the antecubital vein. More preferably the
    Blood collection is done 20 minutes after the ingestion of a nulricional preparation of chemically defined composition. Even more preferably, the nutritional preparation has a caloric concentration of 1'25 kcal / ml, 18'8 9 of proteins, vitamins, minerals and trace elements.
    A fourth aspect of the invention relates to a method for screening, diagnosis, prognosis and monitoring of osteoporosis, comprising steps (a) and (b) as described in the present invention, hereafter referred to as the second method of invention, and also includes:
    c) assign the individual to the group of individuals that present osteoporosis when the result of Formula (1) is greater than 0.5.
    In a preferred embodiment, it refers to the second method of the invention for screening, diagnosis, prognosis and monitoring of osteoporosis.
    In another preferred embodiment, the biological sample isolated in step a) is blood. Preferably the blood is extracted from the antecubital vein. More preferably the blood collection is done 20 minutes after the intake of a nutritional preparation of chemically defined composition. Even more preferably, the nutritional preparation has a caloric concentration of 1.25 kcalfml, 18.8 g of protein, vitamins, minerals and trace elements.
    An individual or "subject" as used herein refers to a mammal, human or non-human, under observation, and more preferably a human being. The individual can be anyone, an individual predisposed to a disease (for example Pea) or an individual suffering from said disease.
    The term "quantify" as used in the description refers to, but is not limited to, the determination of the absolute or relative amount of the markers, their concentration in blood or plasma, as well as any other value or for meter related to them or that may be derived from them.These values or parameters comprise values of signal intensity obtained from any of the physical or chemical properties of said expression products obtained by direct measurement. they include all those obtained by indirect measurement, for example, any of the measurement systems described elsewhere in this document.
    The term uZ-score, Z lumbar score or Zscorelumbar "is the number of standard deviations of a patient with average bone mineral density (OPH) different from the average DPH by age, sex, ethnicity. This value is used in premenopausal women , men under 50, and in children.It also serves to establish if the patient has such a low DPH with respect to his age group that he presumes some secondary cause.This value is the value obtained by means of Bone Densitometry (DEXA).
    The term "logistic regression model, logistic model. Logit model, or maximum entropy classifier" refers to a type of regression analysis, which uses formula 1, used to predict the outcome of a categorical variable (a variable that it can adopt a limited number of categories) depending on the independent or predictive variables. It is useful for modeling the probability of an event occurring as a function of other factors.
    A "biological sample" as defined herein is a small part of a subject, representative of the whole and may be a biopsy or a sample of body fluid. Biopsies are small pieces of tissue and can be fresh, frozen or fixed, as fixed with formalin and embedded in paraffin (formalin · fixed and paraffin embedded FFPE). Samples of body fluids may be blood, plasma, serum, urine, sputum, cerebrospinal fluid, milk or ductal fluid samples and may also be fresh, frozen or fixed. Samples can be surgically removed, by extraction, that is, with hypodermic or other needles, by microdissection or laser capture. The sample must contain any biological material suitable for detecting the desired marker, biomarker or biomarkers / s, therefore, said sample must advantageously comprise material from the subject's cells. Therefore, in a particular embodiment, the sample is a body fluid sample such as a blood or urine sample; preferably, the sample is from blood.
    A "reference sample", as used herein, means a sample obtained from individuals, preferably two or more individuals, known to be free from the disease (osteoporosis) and / or, alternatively, from the healthy population. . Suitable reference levels of the markers can be determined by measuring the levels of said markers in various suitable individuals, and such reference levels can be adjusted for populations of specific individuals or subjects. In a preferred embodiment, the reference sample is obtained from a group of healthy individuals or subjects or from subjects without a previous history of osteoporosis. The profile of marker levels in the reference sample can preferably be generated from a population of two or more individuals; for example, the population may comprise 3, 4, 5, 10, 15, 20, 30, 40, 50 or more individuals or subjects. In another preferred embodiment, the healthy reference population comprises 58 individuals or subjects. In another preferred embodiment, the population with a previous history of reference osteoporosis comprises 58 individuals or subjects.
    The determination of the markers can be done by any means known to the person skilled in the art. In a preferred embodiment, the determinations have been made by luminescence methods.
    s A nutritional preparation, as referred to in the present invention, refers to the intake of a chemically defined composition with a caloric concentration of 1.25 kcal / ml, 18.8 g of proteins, vitamins, minerals and trace elements. , used to improve the determination of blood markers of individuals.
    MEDICAL USES
    10 A fifth aspect of the invention relates to the use of the composition of the invention for the prevention, relief, improvement and / or treatment of individuals selected by the methods of the invention.
    KIT OR DEVICES AND USES
    A sixth aspect of the invention relates to a kit or device, hereinafter kit or device of the invention, comprising a:
    fifteen component A, formed by a blood collector, and / or
    component 8, consisting of one or more blood sample tubes with 10 IJL of a DPP-4 inhibitor per mL of blood, a 96-well plate, conjugated magnetic beads, specific antibodies, standards, buffers, solvents, streptavidin -PE, filter plate, flat bottom plate, sealing tape, and / or
    twenty component e, formed one or more blood sample tubes, tripotassium EOTA, sodium heparin, and / or
    component D, formed by a light spectrum measuring device, and / or
    component E, formed by a densitometer, and / or
    or any of its combinations.
    25 On the other hand, the kit can include all the supports and containers necessary for its start-up and optimization. Preferably, the kit further comprises instructions for carrying out the methods of the invention.
    The term - bone densitometry or OEXN refers to a test to determine bone mineral density. It can be done with X-rays, ultrasound or radioactive isotopes. It serves to
    S the diagnosis of osteoporosis. The test is performed with the device that measures the images and gives a figure of the bone mineral amount per surface. The test works by measuring a specific bone, or more, usually of the spine, hip, forearm. The density of these bones is compared with an average value based on age, sex, size. The comparison of results is used to determine the risk of fractures and the state of osteoporosis in an individual.
    10 A seventh aspect of the invention relates to the use of the kit or device of the invention, for obtaining useful data for the screening, diagnosis, prognosis and monitoring of osteoporosis.
    METHODS IMPLEMENTED BY COMPUTER
    lS The invention also extends to computer programs adapted so that any processing means can implement the methods of the invention. Such programs may have the form of source code, object code, an intermediate source of code and object code, for example, as in partially compiled form, or in any other form suitable for use in the implementation of the processes according to the invention . Computer programs also cover cloud applications based on that procedure.
    twenty An eighth aspect of the invention relates to a computer program comprising program instructions for making a computer carry out the process according to any of the methods of the invention.
    25 In particular, the invention encompasses computer programs arranged on or within a carrier. The carrier can be any entity or device capable of supporting the program. When the program is incorporated into a signal that can be directly transported by a cable or other device or medium, the carrier may be constituted by said cable or other device or means. As a variant, the carrier could be an integrated circuit in which the program is included, the integrated circuit being adapted to execute, or to be used in the execution of the corresponding processes.
    30 For example, the programs could be incorporated into a storage medium, such as a ROM, a CD ROM or a semiconductor ROM, a USB memory, or a magnetic recording medium such as a floppy disk or a HDD. Alternatively, the programs could be supported in a
    transmissible carrier signal. For example, it could be an electrical or optical signal that could be transported through an electrical or optical cable, by radio or by any other means.
    A ninth aspect of the invention relates to a computer-readable storage medium comprising program instructions capable of causing a computer to carry out the steps of any of the methods of the invention.
    A tenth aspect of the invention relates to a transmissible signal comprising program instructions capable of having a computer perform the steps of any of the methods of the invention.
    Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and drawings are provided by way of illustration, and are not intended to be limiting of the present invention,
    EXAMPLES OF THE INVENTION
    In osteoporotic patients, the levels of intestinal peptides, mainly GLp · 1, GLP · 2, or GIP are decreased compared to patients without osteoporosis of the same age and sex. That is why the determination of the levels of these markers, as well as the OPP-4 marker, could be a good strategy to find new ways to screen, diagnose, prognosis and / or monitor osteoporosis.
    Likewise, preparations analogous to intestinal peptides GLp · 1, GLp · 2, GIP, as well as the enzyme responsible for their metabolization, OPP · 4, together with amylin can be used as a treatment for osteoporosis.
    To test this hypothesis, the project was currently being developed.
    In this work, a case-control study was applied, matched 1: 1 by sex and age (+/1 year) to look for potential markers, mainly the intestinal peptides GLP-1, GLP · 2 and GIP, as well as the enzyme responsible for its metabolism, OPP-4, and its relationship with osteoporosis, which may help to understand the pathological processes related to this pathology.
    Likewise, the relationship of the following markers and their association with osteoporosis were analyzed in a more particular way:
    Intestinal peptides Peptide YY and amylin.The blood parameters involved in bone remodeling and their associationwith osteoporosis and with the intestinal peptides GLP · 1, GLP · 2, GIP, peptide YY,amylin and DPP-4 enzyme, especially: 25 · 0H vitamin D, calcium, phosphorus,Magnesium, PTH, osteocalcin, aminoterminal procollagen propeptide, andBeta-ctx collagen earboxiterminal telopeptide (CTX-Beta eross-Iaps).Other blood parameters that are related to osteoporosis: blood count,glucose, total cholesterol and fractions, triglycerides and HbA 1 C.Anthropometric parameters: weight, height, body mass index (BMI), andbody compartments: fat free mass, fat mass (measured byimpedance measurement) are associated with the intestinal peptides studied, with theosteoporosis and with the referred parameters of bone remodeling (osteocalcin,aminoterminal procollagen propeptide, CTX · Beta cross · laps).The intake of the main food groups (dairy, vegetables and vegetables,legumes, fruits, meats, fish and eggs, oils) and their association withosteoporosis and with the peptides referenced above: GLP · 1, GLP · 2, GIP,YY peptide, amylin and DPP-4 enzyme.The intake of macronutrients (proteins, lipids, carbohydrates) and ofmicro nutrients (calcium, zinc, vitamin C, vitamin 812, phallic acid, vitamin 86,vitamin O) studying its relationship with osteoporosis and with peptidesIntestinal: GLP-1, GLP-2, GIP, YY peptide, amylin and DPP-4 enzyme.Physical activity, possible pharmacological treatments and their association withintestinal peptides GLP · 1, GLP · 2, GIP, peptide YY, amylin and the enzyme OPP · 4 andwith osteoporosis
    The population of the study in this work corresponds to the area of influence of the Jaén Hospital Complex.
    Example 1. Study design
    As mentioned above, this work is based on a case-control study, paired 1: 1 by sex and age (+/1 year). It has been considered as exposure to have decreased levels of intestinal peptides, and as an effect to present osteoporosis.
    Sample size and sampling procedure
    According to the results shown in Wojcik et al 2011 (2011), considering a 5% error, a power of 90% and a 10% loss, to detect differences in the contrast of the null hypothesis HO: IJ, = 1J2 by a bilateral T ~ Student Test for two independent samples, and assuming that the average of the Case group is 92.20 units, the average of the Controls group is 106.30 units and the standard deviation of both groups is 12.00 units, 38 patients would be needed in the study.
    In order to bridge the differences between the Wojcik study and the present one and to be able to carry out the statistical analyzes presented with sufficient power, it has been decided to expand the sample to a total of 58 patients per group (cases and control), in total 116 patients aged 18-65 years from the area of influence of the Jaén Hospital Complex. As it is a non-invasive study, this sample expansion does not present any ethical conflict.
    The sampling procedure used will be consecutive to the identification of the patients, who underwent an interview and the review of their medical histories.
    The exclusion criteria that have been marked for both groups are the following:
    Diabetes mellitus (determined by measuring HbA1C).Diagnosis of secondary osteoporosis.Diseases related to calcium metabolism: hypo orhyperparathyroidism, hyperphosphatemia, subclinical and clinical hyperthyroidism.Cushing's diseaseGestation.Previous hospital admission in the last 6 months.Diagnosis of severe cancer disease at the time of inclusion in thestudy.Ileocolic disease: inflammatory bowel disease, intestinal malabsorption,resection and intestinal fistulas.Diagnosis of stage 4 chronic renal failure (GFR <30).In active treatment with: biological factors; GLP-1 analogs; inhibitors ofDDP ~ 4; cholestyramine; Antibodies: phenytoin, phenobarbital, carbamazepine orvalproate; rifampicin; antacids: aluminum salts; antineoplastic: cyclophosphamide;bisphosphonates; strontium ranelate teriparatide; sodium fluoride; corticosteroids; and therapyhormone replacement.
    They will be selected consecutively to their identification among patients who are diagnosed with primary osteoporosis in the Rheumatology service of the Jaén Hospital Complex and who meet the following selection criteria.
    Be diagnosed with osteoporosis by densitometry (dual absorptiometry byX-ray (DEXA)).Accept participation in the study by signing the consentinformed.
    Controls
    Controls or healthy patients were recruited in the family environment of those included in the case group, matched 1: 1 by age and sex with the control group and also:
    They have no criteria for osteopenia or osteoporosis in DEXAThey have not presented a fragility fracture.Accept participation in the study by signing the consentinformed.
    Example 2. Method and Data Collection
    All patients were subjected to a series of tests to determine the markers set forth above:
    Bone Mass Determination
    It has been made using a GE HC brand densitometer, model LUNAR OPX. After weighing and carving the patient, the lumbar spine measurement was performed: specifically the vertebrae from L 1-L4 were analyzed, and the femoral neck was analyzed at the hip. The results were compared with those considered normal according to age and sex.
    The levels or cut-off points of BMD measured by DXA in lumbar spine and hip have been established as:
    Normal: when BMD values are greater than -1 SD in relation to the average of adults (T-score> -1).
    s Osteopenia or low bone mass: BMD values between -1 and -2.5 SD (T-score between -1 and -2.5). Osteoporosis: BMD values <-2.5 SD (Tscore <-2.5), and osteoporosis established when it is associated with one or more osteoporotic fractures3. (WHO Scientific Group 2007).
    Blood determinations
    Two blood extractions were performed:
    10 Basal: for the determination of parameters of bone metabolism and general biochemistry. Postprandial: 20 minutes after the intake of a nutritional preparation with chemically defined formulation for the determination of GLP-1, GLP-2, amylin, VY peptides, as well as the activity of the DPP-4 enzyme.
    lS The following determinations were made in the Clinical Analysis laboratory of the Jaén Hospital Complex:
    twenty General: blood count; biochemistry: glucose; HBA1C; Urea; creatinine; total cholesterol and fractions; calcium; match; and magnesium 6seo metabolism and remodeling parameters: 25-0H vitamin D, PTH, osteocalcin, aminoterminal procollagen propeptide, CTX-Beta cross-laps. These parameters were determined at baseline conditions after fasting for at least 8 h.
    2S 30 At the University of Jaén in the department of Physiology, the GLP-1, GLP-2 determinations were made. GIP, YY psype, amylin and DPP-4. The determination of intestinal peptides will be carried out in plasma samples obtained from blood extracted 20 minutes after the ingestion of a nutritional preparation with chemically defined composition (Resource protein®), of the antecubital vein, the tubes being placed on ice and centrifuged immediately at 4 degrees Celsius, and at 3000 xg, 30 minutes, then immediately stored in a freezer at -80 degrees Celsius. Before blood collection and only in the tube with the blood sample corresponding to the study of the peptides, 10 IJL of an OPP-4 inhibitor (DPP4IDPP4-010, Linco Research Inc, St Charles, Missouri, USA) will be added for each mL of blood, following the technique indicated in Hattori et al (2013). For determinations,
    S 10 will use the 8io-Plex Pro Human Diabetes 10-Plex Assay Kit, from 8io-Rad, specifically designed to determine the levels of GLP-1, GLP-2, GIP, YY peptide and amylin, using Luminex x-MAP technology technology. These determinations will be carried out in the technical research services of the University of Jaén. DPP4 activity (Dipeptidyl aminopeptidasa-4) was determined by fluorimetric techniques using the Sigma-Aldrich -DPP4 Activity Assay Kit ". This assay is based on hydrolysis by the enzyme of the H-Gly-Pro-AMe Substrate enzyme, releasing the product 7-Amino-4-Methyleoumarin (AMe) whose fluorescence will be measured at an excitation of 360 nm and an emission of 460 nm after incubation at 37 ° C. The activity will be expressed as JJmol of AMC per minute and per minute. of plasma These analyzes were carried out in the laboratories of the Physiology area of the University of Jaén.
    Anthropometric determinations and bioimpedance
    A series of parameter determinations were made, according to:
    fifteen Weight, size, waist circumference (cm), and Body Mass Index (BMI). Body behaviors: were carried out by means of an analyzer of the body composition by impedance, Tanita brand, model Me-180MA, and it was determined: Segmental Multifrequency. Total and segmental fat mass (%), Total and segmental fat free mass (%), and Total and segmental muscle mass (%).
    Food survey and physical activity data
    20 2S A semi-quantitative food consumption frequency questionnaire adapted to the Spanish population (Martín-Moreno et al 2003) was used, complemented with the questionnaire used in the study of the SUN cohort, assessing physical activity in METs, (Martínez-González el 2003) . Information on the frequency of consumption is collected based on the type of food (daily, weekly, monthly, annual) of the previous year. The intake per day of energy (Kcalldia) macro nutrients (carbohydrates, proteins, lipids) in grams / day, and micronutrients will be determined: calcium, phosphorus, zinc, vitamin O, vitamin e, vitamin 812 and folic acid.
    Example 3. Variables of the study
    The variables that will be taken into account in the case-control study are:
    30 Sociodemographic: Age (years); Sex (man, (domicile, institution) Dependent Variable: Osteoporosis: Yes / No.woman);Homehabitual
    31
    Analytical Determinations:
    • Specific:
    or Intestinal peptides: GLP-l (pg / ml), GLP-2 (pg / ml), GIP (pg / ml), Amylin (pg / ml), YY Peptide (pg / ml), DPP-4 Activity : The
    activity will be expressed as IJmol of AMC per minute and per mL of plasma.
    o Parameters of bone metabolism and remodeling: PTH (pg / ml), 25-0H serum vitamin O (ng / ml), Osteocalcin (ng / ml), carboxyterminal carboxyterminal peptide Beta-cross laps (CTX) (ng / ml) , Aminoterminal propeptide of total procollagen type lIP1 NP (ng / ml)
    (propeptide).
    • General:
    o Blood count: hematies, leukocytes, platelets (units / L).
    o Standard biochemistry: glucose (mg / dl), HbAlc (%), urea (mg / dl),
    creatinine (g / di), cholesterol (total and fractions) (mg / dl), triglycerides
    (mg / dl), electrolytes (mEq / L) (sodium, potassium, chlorine).
    o Serum calcium (mg / dl).
    o Serum phosphorus (mg / dl).
    o Serum magnesium (mg / dl).
    o Serum albumin (g / di).
    • Bone mass:
    o Lumbar spine
    o Hip Anthropometric parameters:
    • Weight (kg)
    • Talia (cm)
    • IMe (kg / m2)
    Body composition parameters:
    • Total and segmental fat mass (%)
    • Total and segmental free fat mass (%)
    • Total and segmental muscle mass (%)
    Food intake:
    • Intake of the main food groups (dairy, vegetables, legumes, fruits, meats, fish and eggs, oils) g / day.
    • Energy (Kcal / day).
    • Macronutrients: carbohydrates (g / day), Proteins (g / day), Lipids (g / day).
    • Micronutrients: Calcium (mg / day), Phosphorus (mg / day), Vitamin O (mg / day), Zinc (mg / day), Vitamin e (mgldia), Vitamin 812 (~ g / day), vitamin 86 (lJg /day)
    • Folic acid (~ g / day).
    s Physical activity: METs / day.Pharmacological treatments
    Example 4. Statistical analysis of the study variables
    The data collected on the independent and dependent variables mentioned in this study were recorded in an anonymized database, built for this purpose and statistically processed using the SPSS v21 statistical program.
    In the first place, a descriptive statistical analysis of each of the variables in the database is carried out, for this in the case of qualitative variables a table of frequencies is presented and as a graphical representation the graph of sectors. In the case of
    The quantitative variables will present measures of central tendency and as a graphical representation the box and mustache diagram.
    Below is a bivariate analysis to evaluate the different objectives.
    In this way, to check the relationship between intestinal peptides and the presence of osteoporosis and the rest of its variables, the different tests were used.
    20 In the case of a dichotomous qualitative variable and a quantitative variable, the t-$ tudent test for independent samples or the non-parametric U-Mann Whitney test, if the qualitative variable has 3 or more modalities, an ANOVA or Kruskal Wallis non-parametric test, in the case of giving significant results, the corresponding multiple comparisons were studied.
    2S In the case of qualitative variables, the Chi-square test (in r x s tables) or the Fisher test (in 2x2 tables), evaluating the Odds Ratio and their corresponding confidence intervals. If both variables are quantitative the Pearson correlation coefficient, or Spearman
    (according to the normality of the variables).
    The study of the normality of the quantitative variables was carried out through the Kolmogorov Smirnov test or Shapiro Wilks test, depending on the sample size.
    In the multivariable analysis, logistic regression models will be used to determine which variables are associated with the presence or not of osteoporosis. Confusion factors were considered to be those that are not intermediate variables between exposure and effect, and maintain a relationship with exposure (GLP-1, GLP-2, GIP) and the effect (osteoporosis). As a pragmatic criterion, the change in the coefficient by 10% or more was used in the detection of a confounding factor.
    A value of 0 = 0.05 was considered significant for all analyzes.
    In the descriptive analysis, level of studies, type of work, history of illness, pharmacological treatments, nutritional intake, physical activity were assessed. In the mean difference test, no significant differences were found between cases and controls in type of work, history of disease or pharmacological treatments, nor in type of feeding, which indicates a great homegeneity between both groups. There are no significant differences in GLP-1 and GLP-2 between cases and controls in the mean difference test (t-test).
    Nor have significant differences been found in the general analyzes of biochemical and hematimetric parameters between cases and controls in the mean difference test.
    Yes, statistically significant differences were found between cases and controls in relation to anthropometric parameters (weight, body mass index) and in body compartments measured by bioimpedance.
    In relation to the results obtained in the values of the intestinal peptides, and although
    or a significant correlation was found in GLP-1 and GLP-2 between cases and controls in the mean difference test (t-test), a significant association was found in linear regression analysis between GLP-1 and GLP- 2 And the T-score and lumbar Z-score.
    Thus it has been seen that GLP-1 is negatively related to the T-score and Z-score, at the limit of significance in the case group (p = 0.056 and 0.064 respectively) and statistically significant in the controls ( p = 0.015 and 0.010 respectively), while GLP-2 has a positive association with the lumbar T-score in the case group (p = 0.029).
    In the multivariable conditional logistic regression analysis, adjusting for different confounding factors, GLP-1 shows a significant decrease in the risk of osteoporosis (OR = 0.89, 95% confidence interval (0.81 -0.98 ».
    S In relation to OPP · 4 activity, there is a significant correlation of OPP · 4 with bone remodeling markers in the linear regression analysis: a positive and significant association of OPP-4 activity is shown in the case group between DPP-4 activity and osteoealein (p = 0.006), propeptide (p = a, Ot2), and CTX (p = 0.007), while in the control group, it only shows significant association with the activity of the DPP-4, the propeptide (p = 0.022).
    No relationship was found between the YY peptide and osteoporosis.
    The results regarding statistics. to amylin they have been very heterogeneous and did not admit analysis
    10 There is no relationship between anthropometry and bioimpedance.thePIstudiedYvariablesfromremodeledosseous,
    As expected, there is a significant association between osteoporosis and:
    fifteen Bone mass values measured by OEXA (T · score and Z · score in hip and spine). 6seo remodeling markers (osteoealeina, propétidoy CTX). Anthropometry: weight, IMe, and bioimpedance: fat free mass (FFM), total muscle mass (Musc · T) and total fat mass (Fat · T) that is shown in both means difference test and OR analysis.
    In summary, taking into account the most important results, we can say that:
    20 25 It has not been shown that there is a significant decrease in GLP · 1, GLp · 2, PYY, or an increase in OPP4 activity in patients with osteoporosis versus patients without osteoporosis. The GLp · 1 correlates negatively with the T · score and Z · lumbar score mainly in the controls. GLP-2 correlates positively with the lumbar T · score in cases, not controls. DPP4 activity correlates positively and significantly with bone remodeling markers. In multivariable conditional logistic regression, GLP · 1 is associated with a significant decrease in the risk of osteoporosis.
    35
    These results show that there is a relationship of GLP · 1 and GLP · 2 with the bone mass values measured DEXA
    In the case of GLP-1, it has been proven that although the association with the T · score and Z · lumbar score is negative, in logistic regression analyzes, it is verified that GLP · 1 levels are associated with the risk of fracture, that is, higher levels of GLP · 1 decrease the risk of osteoporosis.
    These results, although they may seem contradictory, are in line with other studies that show results for and against the effect of GLP-1 on bone tissue.
    Thus, while Oriessen et al (2015), in a cohort study, show that treatment with GLp · 1 does not affect the risk of fracture, important experimental studies have shown that the use of GLp · receptor agonists 1 (exenatide and liraglutide) improves bone mass in ovariectamized rats, mainly trabecular bone, and increasing the number of osteoclasts, demonstrating that there is a GLp · 1 receptor also in osteoclasts, which could mediate the beneficial effect of GLP receptor agonists -1 on bone tissue, decreasing bone resorption (Pereira et al 2015). Previously, it had been shown that GLP-1 receptors exist in adipocytes and osteocytes which would favor the formation of osteoblasts, and in thyroid C cells which would stimulate calcitonin secretion and indirectly inhibit bone resorption, and also in osteoblasts , GLP-1 receptor, which differs from that existing in pancreatic cells (Nuche-Berenguer et al 2010).
    Therefore the different results would be explained because there is no single type of receiver. There are different polymorphisms in the GLP-1 receptor. These different polymorphisms in the GLP · 1 receptor, give rise to dissociated responses (Cantini et al 2016, Luo et al 2016), and possibly would be the cause of responding and non-responding individuals to the action of GLp · 1 on the woven bone.
    Regarding GLp · 2, the results are consistent with those shown in previous studies, mainly those of the Henriksen et al. Group, which show that GLP-2 favors the increase in bone mass (Henriksen, 2003, 200709 Askav-Hansen 2013).
    In relation to the activity of OPP4, the results published so far are contradictory. Monami et al (2011) in a meta-analysis show that treatment with OPP4 inhibitors decreases the risk of fracture. Bunk et al (2012), on the other hand, showed no relationship between treatment with OPP · 4 inhibitors in bone remodeling (Bunck el al 2012).
    More recently, the association of DPP-4 activity has been shown with the risk of osteoporosis (Zheng et al 2015), and with the risk of fracture (Kim et al 2017). The results show that there is a positive and significant relationship between DPP-4 activity with bone remodeling markers: osteocalcin, propeptide and CTX, coinciding with the results of Notsu et al (2016), which also shows an association of bone remodeling markers with DPP-4 (Notsu et al 2016).
    The limitations of the study must be taken into account because it is a cross-sectional study, and a single analytical determination has been made.
    Although it has not been shown that in osteoporotic patients there is a significant decrease in the levels of GLP-1, GLP-2, PYY or increased DPP4 activity, it has been shown that:
    GLP-2 is positively related to lumbar spine MO in patients withosteoporosis.GLP-1 decreases the risk of osteoporosis.The activity of DPP4 is positively and significantly related to thebone remodeling markers.More studies are needed to deepen the relationship between IP and tissueosseous.
    Example 5. Multivariate logistic regression model for the classification of patients in cases and controls
    A multivariate model is proposed, the dependent variable being the one that measures whether patients are cases or controls, and as independent variables GLP-1, CTX and lumbar Z-score.
    The model includes information on 102 patients, since 19 have lost values.
    The omnibus tests on the model coefficients indicate that the calculated model is statistically significant, p_value = O, OOO (table 1).
    Table 1. Bus tests on model coefficients
    Chi squared glI follow
    Step 1 Step Block Model120,913 120,913 120,9133 3 3, 000, 000, 000
    The coefficient R squared of Cox and Snell indicates that the model explains 69.4% of the variability of the dependent variable, which shows that the model is very suitable.
    5 The Hosmer and Lemeshow Test verifies that the classification that makes the model of patients to osteoporosis does not differ significantly from reality, which means that the model classifies correctly, p_value = 1, OOO.
    Table 2. Hosmer and Lemeshow test
    He passed Chi squaredglI follow
    one , 54581,000
    In the following table (table 3) it can be verified that the observed and expected frequencies do not differ slightly.
    Table 3. Contingency table for the Hosmer and Lemeshow test
    CASCON CONTROL HELMET N = CASETotal
    Observed Expected ObservedExpected
    one 1010,000OR, 00010
    2 109,994OR, 00610
    3 109,962OR, 03810
    4 109,714OR, 28610
    5 88,34121,65910
    Step 1
    6 21,87788,12310
    7 OR, 086109,91410
    8 OR, 022109,97810
    9 OR, 003109,99710
    10 OR, 0001212,00012
    The model correctly classifies 97.1% of cases (table 4), also has high sensitivity, specificity, VPN and vpp values (table 5).
    Table 4. Classification table
    Forecasted
    Observed CASCON Percentage
    CONTROL CASE correct
    Step 1 CONTROL 49 1 98.0 CASCON CASE 2 50 96.2
    Overall percentage 97.1
    Table 5. Table of sensitivity, specificity, NPV and PPV values.
    IC
    Value Lim. InlLim. His p
    VPN 98.0093.12100.00
    VPP 96.1589.97100.00
    Sensitivity 98.0493.25100.00
    Specificity 96.0889.77100.00
    s
    According to this information it can be verified that the fit of the calculated model is adequate.
    Finally, in the model obtained it can be seen that both GLP-1 and ZSCORCOLUM are
    10 statistically significant variables in the model (p_value = 0.009 and p_value = 0.001 respectively), while the CTX variable is not significant but remains in the model due to its importance in the study (table 6).
    Table 6. Variables in the equation.
    B E.T.WaldglI followExp (B)I.C. 95% for EXP (B)
    lower Higher
    GLP1 -, 068, 0266,853one, 009, 934, 888, 983
    He passed CTX-, 7724,286, 032one, 857, 462, 0002053,844
    l ' ZSCORCOLUM-4,4181,27312,033one, 001, 012, 001, 146
    Constant 9,0163,9845,122one, 0248229,918
    Therefore, an equation is finally obtained to classify patients at risk of osteoporosis:
    Probability of OSleoporosis = 1+ /, 0.068 'GLP1. (), 712'crL, 418' z.scORELUMBAR + 9.016)
    5 Thus, when a new patient is evaluated, if the result of the equation is greater than 0.5, it will be classified as an osteoporotic patient. While if the result is less than 0.5 it will be classified as a non-osteoporotic patient.
    To determine the predictive capacity of the model, the ROC Curve is calculated with the predicted probabilities of the model and the original dependent variable. The
    10 results indicate that the area under the ROe curve is 0.994, that is, the predictive capacity of the model is 99.4%, so the model has a very high predictive capacity (Figure 1).
    Finally, as the proposed regression study includes quantitative variables, a linearity study has been carried out in ellogi1. Through the correlation coefficients of
    Pearson shows that such linearity exists and is statistically significant (Table 7).
    Table 6. Correlations.
    GLP1 CTXZSCORCOLUM
    LOGIT_P Pearson correlation-, 211, 431 ..-, 971 ..
    Sig o (bilateral) , 033, 000, 000
    N 102102102
    REFERENCES
    Alvarez-Nebreda ML, Jiménez AB, Rodríguez P, Serra JA. Epidemiology of hip fracture in 5 the elderly in Spain. Bone 200B Feb; 42 (2): 27B-B5.
    Askov-Hansen C, Jeppesen PB, Lund P, Hartmann B, Holst JJ, Henriksen OB. Effect of glucagon-like peptide-2 exposure on bone resorption: Effectiveness of high concentration versus prolonged exposure. Regul Pept. 2013 Feb 10; 181: 4-8
    Baim S, Binkley N, Bilezikian JP, KendJer ol, Hans oB, Lewiecki EM, Silverman S. Official 10 Positions of the International Society for Clinical Oensitometry and Executive Summary of the 2007 ISCo Position Oevelopment Conference. J Clin oensitom. 2008; 11: 75-91.
    Baró F, Cano A, Sánchez Borrego R, Ferrer J, González Rodríguez SP, Neyro JL, Rodríguez Bueno E, Sancho C, Inaraja V, Fernández C, Corral C; FROSPE StudyGroup. Frequency 01 FRAX risk factors in osteopenic postmenopausal women with and without history af fragility
    15 fracture Menapause 2012; 19: 1193-9.
    Bjarnason NH, Henriksen EE, Alexandersen P, Christgau S. Henriksen OB, Christian sen C. Mechanism of circadian variation in bone resorption. Bone 2002; 30: 307-313.
    Bolla9 RJ, Zhong a, Ding KH, Phillips P, Zhong L, ain F, et al. Glucose-dependent insulinotropic peptide is an integrative hormane with osteotropic effects. Mol Cell Endocrine! 20 2001. 25; 177: 35-41.
    42 Sunck MC, Cornér A, Etiasson B, Heine RJ, Shaginian RM, Taskinen MR. Effects of exenatide on measures of ¡3-cell function after 3 years in metformin-treated patients with type 2 diabetes. Careo Diabetes 2011; 34: 2041-7.
    Bunck MC, Poelma M, Eekhoff EM, Schweizer A, Heine RJ, Nijpels G, Foley JE, Diamant M. Effects of vildagliptin on postprandial markers of bone resorption and calcium homeostasis in recently diagnosed, well-control1ed type 2 diabetes patients. J Diabetes. 2012; 4: 181-5.
    Cantini G, Mannucci E, Luconi M. Perspectives in GLP-1 Research: New Targets, New Receptors. Trends Endocrinol Metab. 2016 Jun; 27 (6): 427-38.Clowes JA, Hannon R, Yap T, Hoyle NR, Blumsohn A, Eastell R. Effect of feeding on bone turnover markers and its impact on biological variability of measurement ". Sone 2002; 30: 886-90.
    Clowes JA, Khosla S, Eastell R. Potential role of pancreatic and enteric hormones in regulating bone turnover. J Bone Min Res 2005; 20: 1497-506.
    Oeal C. Future therapeutic targets in osteoporosis. Current Opinion in Rheumatology. 2009. 21; 380-5.
    Oicembrini 1, Mannucci E, Rotella CM. Sone: incretin hormones perceiver or receiver Exp Diabetes Res. 2012; 2012: 519784.
    Oriessen JH, Henry RM, van Onzenoort HA, Lalmohamed A, Burden AM,
    Prieto-Alhambra O, Neef C, Leufkens HG, by Vries F. Sone fracture risk is not associated with the use of glucagon-like peptide-1 receptor agonists: a population-based cohort analysis. Calcif Tissue Int. 2015; 97: 1 04-12
    Gaudin-Audrain C, Irwin N, Mansur S, Flatt PR, Thorens S, Baslé M, Chappard O, Mabilleau
    G. Glucose-dependent insulinotropic po! Ypeptide receptor deficiency leads to modifications of trabecular bane volume and quality in mice. Bane 2013; 53: 221-30.
    Henriksen OS, Alexandersen P, Bjarnason NH, Vilsb011 T, Hartmann S, Henriksen EE, et al. Role of gastrointestinal hormones in postprandial reduction of bane resarption. J Sone Miner Res. 2003; 18: 2180-9.
    Henriksen OS, Alexandersen P, Hartmann B, Adrian CL, Syrjalsen 1. Sone HG, et al. Disassociation of bone resorption and formation by GLP-2: a 14-day study in healthy postmenopausal women. Sane 2007; 40: 723-9.
    Henriksen OS, Alexandersen P, Hartmann 8, Adrian CL, Syrjalsen 1, Sone HG, HoIst JJ, Christiansen C. Four-month treatment with GLP-2 significantly increases hip SMo: a randomized, placebo-controlled, dose-ranging study in postmenopausal women with low BMD. Bone 2009; 45: 833-42.
    S Janssen P, Rotondo A, Mulé F, Tack J. Review article: a comparison of glucagon-like peplides I and 2. Alimenl Pharmaeol Ther. 2013 Jan; 37 (1): 18-36.
    Jódar E. [Characteristics and types of GLP-1 receptor a90nists. individualized Iherapy]. Med Clin (Bare). 2014 Sep; 143 SuppI2: 12-7. Anopportunityfor
    10 Kim H, Baek KH, Lee SY, Ahn SH, Lee SH, Koh JM, Rhee Y, Kim CH, Kim DY, Kang MI, Kim BJ, Min YK. Association of circulating dipeptidyl-peptidase 4 levels with osteoporotic fraelure in poslmenapausal women. Osleoporos Inl. 2017 Mar; 28: 1 099-11 08.
    Labouesse MA, Gertz ER, Pieeolo BD, Souza EC, Sehusler GU, Wilbraehl MG, al. Associations among endocrine, inflammatory, and bone markers, body composition and weight loss induced bone loss. Sane. 2014 Jul; 64: 138-46.
    fifteen Luo G, Liu H, Lu H. Glucagon-like peptide-1 (GLP-1) receptor agonists: potential to reduce fracture risk in diabetic patients Mr J Cl in Pharmacol. 2016 Jan; 81 (1): 78-88.
    Ma X, Meng J, Jia M, et al. Exendin-4, a glucagon-like peptide-1 receptor agonist, prevents osteopenia by promoting bone formation and suppressing bone resorption in aged ovariectomized rats. J. Sone Miner Res. 2013; 28: 1641-52.
    twenty Mabilfeau G, Mieczkowska A, Irwin N, Flatt PR, Chappard D. Optimal bone mechanical and material properties require a functional glucagon-like peptide-1 receptor. J Endocrinol 2013; 219: 59-68.
    Mannucci E, Dicembrini 1. Drugs for type 2 diabetes: role metabolism. Clin Cases Miner Sone Metab. 2015; 12: 130-4. intheregulationofbone
    25 Martin Moreno JM, Boyle P, Weevil L, Maisonneuve P, Fernández Rodriguez JC, Salvini S, Willett WC. 1993. oevelopment and validation of a tood frequency questionnaire in Spain. Int J Epidemiol; 22: 512-9.
    Martinez de Vitoria E, Mañas M, Yago MD. Physiology of Digestion. In Gil A (director). Nutrition Treaty. Madrid 2010. Pan American. 173-201.
    44
    Martínez-González MA, Sánchez-Villegas A, by Irala-Estévez J, Marti A, Martínez JA. 2002. Mediterranean diel and stroke: objectives and design of the SUN Projet. Nutritional Neuroscience; 5: 65-73.
    S Monami M, Dicembrini 1, Antenore A, Mannucci E. Dipeptidyl peptidase-4 inhibitors and bone fractures: a meta-analysis of randomized clinical trials. Diabetes Care 2011; 34: 2474-6
    Nuche-Berenguer 8, Lozano D, Gutiérrez-Rojas 1, Moreno Villanueva-Peñacarrillo ML GLP-1 and exendin-4 can osteopenia. J Endocrinol. 2011 May; 209: 203-10. P, Mariñoso ML, Esbrit P, reverse hyper1ipidic-related
    10 Nuche-Berenguer S, Portal-Núñez S, Moreno P, González N, Acitores A, López-Herradón A, Esbrit P, Valverde 1, Villanueva-PeñacarrilJo ML. Presence of a functional receptor for GLP-1 in osteoblastic cells, independent of the cAMP-linked GLP-1 receptor. J Cell Physiol. 2010 Nov; 225 (2): 585-92.
    fifteen Nuche-Berenguer 8, Moreno P, Esbrit P, Dapia S, Caeiro JR, Cancelas J, Haro-Mora JJ, Villanueva-PeñacarrilJo ML. Effect of GLP-1 treatment on bone turnover in normal, type 2 diabetic, and ¡nsulin-resistant states. Calcif Tissue Int. 2009 Jun; 84 (6): 453-61.
    Notsu M, Kanazawa 1, Tanaka S, Yamaguchi T, Sugimoto T. Serum dipeptidylpeptidase-4 ls associated with multiple vertebral fractures in type 2 diabetes mellitus. Clln Endocrinol (Oxf). 2016 Mar; 84: 332-7
    twenty Pereira M, Jeyabalan J, J0rgensen CS, Hopkinson M, AI-Jazzar A, Roux JP, Chavassieux P, Orriss IR, Cleasby ME, Chenu C. Chronic administration of Glucagon-like peptide-1 receptor agoni5ts improves 1rabecular bone mass and architecture in ovariectomised mice. Sone 2015 Oec; 81: 459-67.
    2S SEOIMM Clinical practice guidelines in postmenopausal, glucocorticoid and male osteoporosis. Spanish Society of Bone Research and Mineral Metabolism. 2014 version. Updated March 2015.
    Tsukiyama K, Yamada Y, Yamada C, Harada N, Kawasaki Y, Ogura M, et al. Gastric inhibitory polypeptide as an endogenous factor promoting new bone formation after food ingestion. Mol Endocrinol. 2006; 20: 1644-51.
    Walsh JS, Henriksen OS. Feeding and bone. Arch Siochem Biophys. 2010. 1; 503: 11-9.
    Four. Five WHO Scientific Group. Assessment of osteoporosis at the primary health care level Geneva
    (Switzerland) World Health Organization; 2007 [updated 2007; cited May 2011].
    Wojcik M, Meenaghan E, Lawson E. Reduced Amylin Levels Are Associated with Low Bone Mineral Density in Women with Anorexia Nervosa. Bone 2010 March; 46: 796-800. 5 Yavropoulou MP, Yovos JG. Incretins and bone: evolving concepts in nutrient-dependent
    regulation 01 bone turnover. Hormones (Athens). 2013; 12: 214-23.Zhong a, Itokawa T,
    Sridhar S, Ding KH, Xie D, Kang B, 801la9 WB, et al. Effects of glucose-dependent insulinotropic peptide on osteoclast function. Am J Physiol Endocrinol Metab. 2007;
    292: E543-8.
    10 Zheng T, Yang L, Liu Y, Liu H, Yu J, Zhang X, ain S. Plasma OPP4 Activities Are Associated
    With Osteoporosis in Postmenopausal Women With Normal Glucose Tolerance. J Clin
    Endocrinol Metab. 2015 Oct; 1 00 (1 0): 3862-70.
    权利要求:
    Claims (18)
    [1]
    1. Use of a composition comprising glucagon-like peptide 1 (GLP-1), GLP-1 analogs, GLP-1 receptor agonists, peptide similar to
    5 glucagon 2 (GLP-2), GLP-2 analogs, GLP-2 receptor agonists,OPP-4 inhibitors, or any of their salts, esters, tautomers, solvates andpharmaceutically acceptable hydrates, or any combination thereof, in thedevelopment of a medicine for the prevention, relief, improvement and / or treatment ofosteoporosis
    2. Use of the composition according to the preceding claim further comprising GIP, YV peptide, amylin or any of its pharmaceutically acceptable salts, esters, tautomers, solvates and hydrates.
    [3]
    3. Use of the composition according to any of claims 1-2, wherein the GLP-1 receptor analogs and agonists are selected from the list that
    15 consists of repaglinide, nateglinide, exenatide, pramlintide, liraglutide, lixisenatide, albiglutide, dulaglutide or any combination thereof.
    [4]
    Four. Use of the composition according to any of claims 1-3, wherein the GLP-2 analog consists of teduglutide.
    [5]
    5. Use of the composition according to any of claims 1-4, wherein
    20 DPP-4 inhibitors are selected from the list consisting of sitagliptin, vildagliptin, saxagliptin, alogliptin, linagliptin, gemigliptin, sitagliptin and simvastatin, or any combination thereof.
    [6]
    6. Use of the composition according to any of claims 1-5, wherein the composition further comprises another active ingredient, preferably an agent
    25 which affects bone structure and mineralization that is selected from the list consisting of bisphosphonates, bone morphogenic proteins, strontium ranelate, denosumab, estrogens, estrogen receptor stimulants, raloxifene, teriparatide, parathyroid hormone, calcitonin, calcium, cholecalciferol , calcifediol or any combination thereof.
    7. Use of the composition according to any of claims 1-6, wherein the composition further comprises one or more pharmaceutically acceptable excipients.

    [8]
    8. Use of glucagon-like peptide markers (GLP-1), carboxyterminal telopeptide of Beta-ctx collagen (CTX-Beta cross-Iaps), or their combination for the screening, diagnosis, prognosis and / or monitoring of osteoporosis.
    [9]
    9. The use of the markers according to the preceding claim further comprising the glucagon-like peptide 2 (GLP-2), dipeptidyl peptidase 4 (OPP-4), or any combination thereof.
    [10]
    10. The use of the markers according to any of claims 8 · 9, further comprising the gastric inhibitor pOpeptide (GIP), YY peptide, amylin, or any combination thereof.
    [11 ]
    eleven . The use of the markers according to any of claims 8-10, which further comprises 25-0H vitamin O, calcium, phosphorus, magnesium, parathyroid hormone (PTH), or any combination thereof.
    [12]
    12. The use of the markers according to any one of claims 8-11, further comprising osteocalcin, aminoterminal procollagen propeptide, beta-ctx collagen carboxyterminal telopeptide (CTX-Beta cross-Iaps), or any combination thereof.
    [13]
    13. A method of obtaining useful data for the screening, diagnosis, prognosis and / or monitoring of osteoporosis, comprising:
    a) quantifying the expression products GLP-1, GLP-2, DPP-4 and CTX-Beta crosslaps, as defined in any of claims 8-12, in an isolated biological sample of an individual who has been previously administered a nutritional preparation, and Zscorelumbar, and
    b) calculate the probability of an individual suffering from osteoporosis based on formula (1):
    Probability
    1+ e (-O, 068 ° GLP1-O.77PCTX-4,4LS "ZSCORElUMBAR.,. 9.0l6J
    Formula 1)
    where
    GLP1: is the value of the peptide similar to glucagon 1 (GLP-1);

    CTX: is the value of the 6seo CTX Beta-cross laps remodeling parameter; Y
    Zscorelumbar: is the value of the lumbar Z score obtained by bone Oensitometry (DEXA).
    [14]
    14. A method for screening, diagnosis, prognosis and / or monitoring of osteoporosis,
    5 comprising steps (a) and (b) according to the preceding claim, and further comprising:
    e) assign the individual to the group of individuals presenting osteoporosis when the result of Formula I is greater than 0.5.
    [15]
    15. The method according to any of claims 13-14, wherein the biological sample isolated in step a) is blood.
    [16]
    16. Use of the composition according to any of claims 1 ~ 7, for the prevention, relief, improvement and treatment of the individuals selected by the method according to any of claims 13-15.
    [17]
    17. A kit or device comprising a:
    Component A, connected by a blood collector, and / or
    component B, consisting of one or more blood sample tubes with 10 IJL of an OPP-4 inhibitor per mL of blood, a 96-well plate, conjugated magnetic microspheres, specific antibodies, standards, buffers, solvents, streptavidin -PE, filter plate, flat bottom plate, sealing tape, and it
    20 component C, treated by tripotassium EDTA, sodium heparin, and so
    component O, formed by a light spectrum measuring device, and
    component E, formed by a densitometer, and
    or any of its combinations.
    [18]
    18. Use of the kit or device according to claim 17, for obtaining useful data for the screening, diagnosis, prognosis and / or monitoring of osteoporosis.
    [19]
    19. A computer program comprising program instructions to make a computer practice the method as described in any of claims 13-15.
    [20]
    twenty. A computer-readable storage medium comprising program instructions capable of having a computer perform the steps of any of the methods described in any of the claims.

    13-15
    S 21. A transmissible signal comprising program instructions capable of having a computer perform the steps of any of the methods described in any of claims 13-15.
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    同族专利:
    公开号 | 公开日
    ES2687220B1|2019-08-08|
    WO2018172590A1|2018-09-27|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    ES2160985T3|1996-12-09|2001-11-16|Osteometer Biotech As|SANDWICH TYPE TESTS FOR COLLAGEN FRAGMENTS.|
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