Method for determining miacalcic preparation dose

FIELD: medicine.

SUBSTANCE: method involves receiving mononuclear cells from 6-8 ml of patient blood by adhesion in control and case tablet cells. 9 mcl of Miacalcic solution of 500 IU/l are added into case cell and the same volume of solvent is added to control cell. Culturing begins. NO3 level increasing in Miacalcic medium to 10%, Miacalcic dose of 50 IU/day is to be prescribed. The value being equal to 10-30%, 100 IU/day is to be prescribed. The value being greater than 30%, 200 IU/day is to be prescribed.

EFFECT: enabled differential approach to treating osteoporosis cases.

1 tbl

 

The invention relates to medicine, more specifically to Orthopaedics and rheumatology, and is intended for differentiated assignments and individual dosage of synthetic calcitonin salmon (miacalcin) in patients with osteoporosis.

Drugs from the group of calcitonin used in complex treatment of patients with osteoporosis. Combined with drugs from other groups, the most widespread synthetic calcitonin salmon produced by a Swiss company Novartis under the name "Miacalcic". The mechanism of developing osteoporosis depends on the predominant causes and individually in each patient. Therefore, the development of new ways of organising differentiated and individual dispensing of medicines affecting bone tissue, plays an important role.

The most effective methods of application of drugs from the group of calcitonin widely entered into the practice of treating patients with osteoporosis, consider the recommended Novartis dosing schedules of miacalcic for the treatment of patients with episodes of acute or chronic back pain. These methods are used as follows: during the episode of acute back pain appoint 50-100 ED drug subcutaneously or intramuscularly for 2 weeks, then 50 UNITS a day up to 2 months, training for 2 months 3 p is for a year, chronic back pain appoint 50 IU intramuscularly every other day, then 50 IU three times per week up to 2 months, training for 2 month 3 times per year with a 2 months break.

However, these methods of application of miacalcic only take into account the nature of the pain syndrome. It is known that osteoporosis has many etiopathogenetic variants and clinical efficacy of miacalcic depends on the leading mechanisms of disease in a specific patient. It is also known that a number of patients when using miacalcic effect is absent or detected intolerance to the drug.

For the effective assignment of miacalcic requires the use of differentiated schemes with a preliminary estimate of drug action in vitro and individual its dosage.

Miakalcik is a synthetic analogue of calcitonin salmon, a peptide hormone secreted parafollicular cells of the thyroid gland. Widely known for its ability to inhibit osteoclastic resorption by suppressing the activity of osteoclasts and decrease their number, gipocalziemiceski effect, analgesic effect, a positive effect on bone formation [3, 4]. It is also widely known patterns of use of this drug in patients with osteoporosis, depending on the nature of the pain syndrome[1, 2, 3, 4]. However, it is not known on a differentiated approach to the appointment of miacalcic and individual dosing on the basis of in vitro tests.

Known ways to spend dosed appointment of miacalcic depending on the nature of the pain syndrome.

Known, in particular, the method of application of miacalcic patients with an episode of acute back pain by assigning 50-100 IU subcutaneously or intramuscularly for 2 weeks, then 50 UNITS a day up to 2 months, training for 2 month 3 times per year with a 2 months break.

The disadvantages of this method include that it does not allow individual dosing of miacalcic. It is known that osteoporosis has many etiopathogenetic variants and clinical efficacy of any drug depends on the leading mechanisms of the disease in each individual patient.

It is established that in the process of treatment of patients with osteoporosis should consider etiopathogenetic variant of the disease and to provide individualized dosing of drugs [4].

In addition to the low efficiency of this method it should be noted the absence of the possibility of a preliminary assessment of the effectiveness of the drug in vitro prior to the appointment of his patient.

The aim of the invention is to increase clinches the th effectiveness of the drug miakalcik by individual dosage.

The goal is to reach what in the way of drug use miakalcik according to the invention is obtained from heparinized peripheral blood of patients with osteoporosis mononuclear cells using gradient centrifugation and evaluate a known manner, the level of production of endogenous nitric oxide mononuclear cells in the presence of miacalcic in vitro. Miakalcik prescribed to patients with higher level of production NO3and the total amount of the final metabolites of nitric oxide in the culture medium in the presence of this drug in vitro. The dose is adjusted depending on the degree of enhancement products NO3: up to 10% - 50 U/day, 10-30% - 100 IU/day, more than 30% - 200 IU/day. The method allows for differentiated application and individual dosage adjustment.

Comparative analysis of stated and known, selected as a prototype, technical solutions showed that the difference of the proposed method is the appointment of miacalcic only patients with increased level of production NO3and the total amount of the final metabolites of nitric oxide in the culture medium in the presence of the drug in vitro, the dose is adjusted individually depending on the degree of enhancement products NO3: up to 10% - 50 U/day, 10-30% - 100 IU/day, more than 30% - 200 IU/day.

The presence of the distinctive features of the prototype characteristics ensures compliance with the proposed method the criterion of "novelty".

The authors first established the possibility of differentiated applications and individual dosage of miacalcic. As it turned out, the need for the appointment of miacalcic depends on its effect in vitro on the production of NO3and the total amount of the final metabolites of nitric oxide in the culture medium of mononuclear cells, and the dose of the drug may individually vary depending on the degree of enhancement products NO3. It has allowed to selectively assign miakalcik sick and individually to dispense the drug in the range from 50 to 200 IU per day.

A significant sign of "assess the level of endogenous production of nitric oxide by mononuclear cells in the presence of miacalcic in vitro in a known manner. Miakalcik prescribed to patients with higher level of production NO3and the total amount of the final metabolites of nitric oxide in the culture medium in the presence of this drug in vitro. The dose is adjusted depending on the degree of enhancement products NO3: up to 10% - 50 U/day, 10-30% - 100 IU/day, more than 30% - 200 IU/day is a new and ensures compliance with the proposed technical solution the criterion of "substantial differences".

The inventive method ASU is estlat as follows.

In a patient with osteoporosis in the morning, on an empty stomach by puncture of the vein in the elbow bend in a sterile tube get 6-8 ml of blood. In the blood in the test tube, add 5-10 IU of heparin. Using gradient centrifugation of heparinized blood is obtained mononuclear cells and carry out the adhesion of mononuclear cells in the wells of sterile tablet. Then prepare a solution of miacalcic in sterile 0.9% sodium chloride solution with a concentration of 500 U/liter. After receipt of adhered cells in the wells with cells and nutrient medium was added 9 μl of a solution of miculescu (experience), and the control hole is equal to the volume of the distribution of the liquid, and then mononuclear cells were cultured for 48 hours at 37°at 5% carbon dioxide. The level of production of endogenous nitric oxide is estimated by the concentration of the final stable metabolites of nitric oxide through reaction Griss. Miakalcik prescribed to patients with higher level of production NO3and the total amount of the final metabolites of nitric oxide in the culture medium in the presence of this drug in vitro and conduct individual selection of doses of the drug depending on the degree of enhancement products NO3: up to 10% - 50 U/day, 10-30% - 100 IU/day, more than 30% - 200 IU/day. Miakalcik in the individual dose is administered subcutaneously or intramuscularly 1 time per day in those who tell 2 weeks then 50 UNITS a day up to 2 months, training for 2 month 3 times per year with a 2 months break.

The authors performed a comprehensive treatment of 14 patients with osteoporosis dosing of miacalcic proposed method. Excellent treatment believed in the complete disappearance of pain and the restoration of motor activity of the patient, good for slight residual pain syndrome and high activity of the patient, satisfactory moderate residual pain syndrome and reduced activity of the patient, and a bad one at the unchanged intensity of pain syndrome and severe restriction of activity of the patient. The results of in vitro tests and treatment of patients presented in the table. It is established that there is a close connection between the influence of miacalcic in vitro on the production of NO3and the total amount of the final metabolites of nitric oxide in the culture medium and the effect of treatment.

The authors found that in the absence of increasing the production of NO3and the total amount of the final metabolites of nitric oxide in the culture medium of mononuclear cells in the presence of miacalcic in vitro, low clinical efficacy of treatment with this drug. Thus, the studies revealed the relationship between the influence of miacalcic products NO3and commanagement end of metabolites of nitric oxide in the culture medium mononuclear cells in vitro and clinical effects of this drug, confirmed immunological method, demonstrates the effectiveness of the proposed method differentiated use and individual dosage of synthetic calcitonin salmon (Miacalcin) in patients with osteoporosis.

Example 1: the Patient-VA AV, 64 years. (Test No. 3) Enrolled in the Department of trauma 01.12.03 with the diagnosis of Postmenopausal osteoporosis, pathologic fracture of the body of L1 vertebra" and intense pain. 03.12.03 in the morning on an empty stomach, the patient is taken venous blood (8 ml, heparinization 10 IU of heparin. After 2 hours the blood delivered in immunological laboratory, where using gradient centrifugation derived mononuclear cells. Next conducted the adhesion of cells in the wells of sterile tablet. After receipt of adhered cells in the wells with cells and culture medium was added 9 μl of a solution of miacalcic in a dilution of 500 U/liter (experience), the control hole is equal to the volume of the distribution of the liquid. As the distribution of the liquid used sterile 0.9% sodium chloride solution. Mononuclear cells cultivated for 48 hours at 37°at 5% carbon dioxide. The level of production of endogenous nitric oxide is evaluated according to the final concentration of stable metabolites of nitric oxide through reaction Griss. It was established that the patient and eat the increased levels of NO production 3and the total amount of the final metabolites of nitric oxide in the culture medium in the presence of miacalcic in vitro (without drug NO317,05 µmol/l, total nitrogen 20,1 µmol/l, in the presence of the drug NO341,7 µmol/l, total nitrogen 44,6 µmol/l). The production level of NO3in the presence of the drug were increased by more than 30%. The patient is assigned miacalcic in the form of intramuscular injection, 1 time per day 200 IU within 14 days. After treatment achieved excellent results: pain controlled completely, the activity of the patient is restored. After discharge from the hospital continued reception of miacalcic outpatient 50 IU intramuscularly every other day up to 2 months.

Example 2: the Patient-VA VE, 72 years. (Test No. 10) Received the Department of trauma 11.03.03 with the diagnosis of Steroid osteoporosis, multiple pathological fractures of the bodies of the thoracic and lumbar vertebrae", severe pain, a sharp limitation of activity 12.03.03 in the morning on an empty stomach, the patient is taken venous blood (8 ml, heparinization 10 IU of heparin. After 2 hours the blood delivered in immunological laboratory, where using gradient centrifugation derived mononuclear cells. Next conducted the adhesion of cells in the wells of sterile tablet. After receipt of adhered cells in the wells with cells and nourishing the environment added 9 μl of a solution of miacalcic in a dilution of 500 U/liter (experience), the control hole is equal to the volume of the distribution of the liquid. As the distribution of the liquid used sterile 0.9% sodium chloride solution. Mononuclear cells cultivated for 48 hours at 37°at 5% carbon dioxide. The level of production of endogenous nitric oxide is evaluated according to the final concentration of stable metabolites of nitric oxide through reaction Griss. It was established that the patient has a reduced level of production of NO3and the total amount of the final metabolites of nitric oxide in the culture medium in the presence of miacalcic in vitro (without drug NO319,33 µmol/l, total nitrogen 21,4 µmol/l, in the presence of the drug NO311.5 µmol/l, total nitrogen of 13.9 µmol/l). The patient is assigned miacalcic accepted way in the form of intramuscular injection, 1 time per day on 100 ED for 14 days. After treatment achieved unsatisfactory results: the intensity of the pain has not changed, the activity of the patient is severely limited, requires nursing care. After discharge from the hospital continued reception of miacalcic outpatient 50 IU intramuscularly every other day up to 2 months. After 2 months remain intense pain, the patient activity remains low. The expediency of application of miacalcic in this case is doubtful.

Example 3: the Patient set E.N., 67. (Test No. 11) Received in the Department of trauma 15.04.03 with the diagnosis of Senile osteoporosis, multiple pathological fractures of the bodies of the thoracic vertebrae", severe pain, significant restriction of activity 17.04.03 in the morning on an empty stomach, the patient is taken venous blood (8 ml, heparinization 10 IU of heparin. After 2 hours the blood delivered in immunological laboratory, where using gradient centrifugation derived mononuclear cells. Next conducted the adhesion of cells in the wells of sterile tablet. After receipt of adhered cells in the wells with cells and culture medium was added 9 μl of a solution of miculescu in a dilution of 500 U/liter (experience), the control hole is equal to the volume of the distribution of the liquid. As the distribution of the liquid used sterile 0.9% sodium chloride solution. Mononuclear cells cultivated for 48 hours at 37°at 5% carbon dioxide. The level of production of endogenous nitric oxide is evaluated according to the final concentration of stable metabolites of nitric oxide through reaction Griss. It was established that the patient has a reduced level of production of NO3and the total amount of the final metabolites of nitric oxide in the culture medium in the presence of miacalcic in vitro (without drug NO3the value of 16,81 µmol/l, total nitrogen and 17.2 µmol/l, in the presence of the drug NO316,32 µmol/l, total nitrogen 16,9 µmol/l). The patient is assigned miacalcic the accepted way in the form of intramuscular injection, 1 time per day on 100 ED for 14 days. After treatment achieved unsatisfactory results: the intensity of the pain has not changed, the activity of the patient is limited, serves himself with difficulty. After discharge from the hospital continued reception of miacalcic outpatient 50 IU intramuscularly every other day up to 2 months. After 2 months is maintained pain syndrome, the patient activity remains low. The expediency of application of miacalcic in this case is doubtful.

Thus, as seen from the above examples, the appointment of miacalcic depending on its influence on the production of NO3and the total amount of the final oxide metabolites in the culture medium mononuclear cells in vitro is effective in improving the quality of treatment of patients with osteoporosis.

Tests of the proposed method confirmed its advantages over the method of the prototype: unlike the prototype method that does not provide a differentiated assignment of miacalcic, offer allows you to use the drug only to patients in whom the level of production of NO3and the total amount of the final oxide metabolites in kulturalny environment mononuclear cells in vitro. Moreover, accounting for the increase in NO production3allows for individual dosage adjustment.

Achievement like what about the results became possible due to set by the authors of the close relationship between the influence of miacalcic products NO 3and the total amount of the final metabolites of nitric oxide in the culture medium mononuclear cells in vitro and clinical effects of this drug, confirmed by immunological method.

Thus, individual dosage adjustment miakalcik increases its clinical efficacy.

Sources of information:

1. Benevolensky LI Osteoporosis: epidemiology, diagnosis. Calcitonin in the treatment of osteoporosis: guidelines for physicians / Liyanawatte, Ehimare, Lyatoshynsky and others - M., 1997. - 32 S.

2. Wertkin A.L. Clinical application of miacalcic in the treatment of postmenopausal osteoporosis / Allwarden, Laulainen, Anticev // Russian medical journal. - 2003. - T, No. 27. - C.2-7.

3. Nasonov EL Place of miacalcic (synthetic calcitonin salmon for the treatment and prevention of osteoporosis / Elenasnow, Iascinina, Wavev etc. // Therapeutic archive. - 1997. No. 7. - Pp.86-89.

4. Rozhen L.Y. Systemic osteoporosis / Lyatoshinsky. - M.: Publisher Mokeev, 2000. - 196 S.

The influence of miacalcic on the production of nitric oxide in vitro and patient outcomes
no testWithout medicationIn the presence of the drugNO2NO3Total NNO2NO3Total N
1.6,340,146,46,843,350,1Good
2.8,525,734,25,833,639,4Good
3.3,0517,0520,12,941,744,6A great price.
4.2,9113,4916,42,3814,8217,2Satis.
5.is 3.0810,8213,92,513,1the 15.6Good
6.9,111,220,38,613,121,7Satis.
7.3,1218,5821,71,2629,1430,4Satis.
8.5,1234,2839,42,7339,0741,8Satis.
9.10,4/td> 31,742,111,135,646,7Good
10.2,0719,3321,42,411,536,1Newdow.
11.0,39the value of 16,8117,20,5816,3216,9Newdow.
128,512,921,49,326,836,1Good
13.5,6the 11.617,2as 4.0212,8816,9Good
14.4,7312,6717,42,89the value of 16,8119,7Good

The method of dispensing of the drug miakalcik in patients with osteoporosis, characterized in that from 6-8 ml of blood of a patient receive mononuclear cells, which desirous in experimental and control wells, then in seasoned well add 9 µl of a solution of miacalcic with a concentration of 500 IU/l and in the control hole is equal to the volume of the distribution of the liquid, cultivate and assess the levels of NO3and when the level of NO3in the presence of miacalcic up to 10% of the patient miacalcic dose of 50 IU/day, up to 10-30% - 100 IU/day, more than 30% - 200 IU/day.



 

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