RU2405559C2 - Method of treating diabetes mellitus - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely, to endocrinology, and deals with treatment of diabetes mellitus. For this purpose into abdomen hypodermic cellular tissue of recipient introduced is injection composition, including bio-degradable gel-containing matrix and cell suspension of primary culture of patient's pancreatic islet tissue.

EFFECT: method ensures normalisation of glucose level in blood for a long time period due to recovery of function of patient's pancreatic islet tissue.

5 cl, 2 tbl

Description

The invention relates to medicine and can be used in the general course of treatment for diabetes.

In connection with the wide spread of diabetes mellitus, the task of finding new effective ways to treat it and prevent complications remains relevant for practical health care. Among these methods prescribed for ineffective or insufficient effective drug therapy, we can mention the transplantation of β-cells of the pancreas. The biomaterial is obtained from newborn rabbits (up to 80 individuals) and, after special processing, dispersion and successive passages, a β-cell culture (2 million) is introduced in the form of a suspension (up to 20.0 ml) under the fascia of the rectus abdominis muscle.

The disadvantages of this method include the mechanical leaching of capable of proliferation of donor material, a change in the amount of donor material at the injection site, an uncontrolled change in glycemia after surgery, the unstable nature of post-transplant glycemia, the risk of hypoglycemic coma in the post-transplant period, the restoration of hyperglycemia 6-8 months after transplantation and high cost of the method.

The objective of the invention is to provide a method for the treatment of patients suffering from type I and type 2 diabetes mellitus, while the technical result achieved is to restore the norm of endocrine relations of the patient’s pancreatic tissue by directed excitation of regulatory systems in his body, as well as to reduce or completely cancel the sugar-lowering drug therapy.

To achieve the result, a method for the treatment of diabetes mellitus is proposed, comprising introducing into the subcutaneous tissue of the abdomen a recipient of an injection composition in the form of a tissue culture and biodegradable gel-containing matrix, the primary culture of islet tissue of the pancreas of the donor in the form of a cell suspension is used as a tissue culture.

Embodiments of the method include the use of a human or animal as a donor, and as a tissue culture, a primary culture of embryonic islet or primary culture of fetal islet tissue of the pancreas.

The principle and ideology of the proposed method are based on the possibilities of correcting the impaired function of organs (in the claimed method - the pancreas) through previously studied and convincingly proven mechanisms for restoring their functions by activating internal reserves. Of course, this is possible if the affected organ has a residual morphological substrate. The application of the proposed method in this case allows not only to hope for the success of the taken medical actions, but also to expect a steady rehabilitation of the lost or leveling of the excess function of the organ damaged by the disease.

Having come into contact with the recipient’s organism, the donor material immediately changes the established balance of the endocrine relationships in the recipient’s blood plasma, since its incretum is absorbed into the blood at the implantation site, spreading its effect by the humoral way, “automatically” through the central nervous system and / or reflexively. The information thus obtained is accepted at the level of the cell membranes of the target organ, the receptor zones of which have a stoichiometric affinity for the regulatory hydrocarbon compounds that make up the hormone. This leads to a change not only in the balance of the “norm of endocrine relationships” in the recipient’s blood plasma, but also in the functional activity of the target organ cells, since they are forced to enter a state of “excitation”.

The biological substance transferred from the donor material to the recipient in a humoral way and, more importantly, transferred from the recipient to the donor material, is a composition of hydrocarbon compounds (proteins, peptides, mucopolysaccharides, etc. - more than 200 items, divided into 24 fractions) molecular weighing from 400 to 400,000 daltons, as well as ions (K ⁺ , Na ⁺ Ca ⁺⁺ , Mg ⁺⁺ , C1 ^- etc.), determines the possibility of the correct selection of the target organ (islets of the recipient's pancreas Langerhans) and restoration his (them) is adequate th function.

The restoration of the function of the organ thus chosen as the target occurs due to the fact that the donor material endowed with the linked properties of tissue and functional specificity has its own hormone, the composition of which is characteristic of healthy tissue. Due to this, the donor material provides not only the choice of the target in the recipient’s body, but also actively intervenes in the established balance of relationships between biologically active substances in the recipient’s blood plasma, assuming the function of the active participant in the general system of neurohumoral regulation, including the function of the organ the target.

The proposed method in practice can be carried out as follows. Immediately prior to surgery, a syringe containing a biodegradable matrix, for example, a heterogeneous implantable Sphero® GEL gel in an amount of 1.0 ÷ 2.0 ml, is placed vertically with the piston down. The syringe piston is withdrawn, a micropipette tip is introduced through the syringe connector into the vacant volume and a suspension of xenogenic biomaterial is injected in the form of a culture of authentic xenogenic donor tissue (according to the claimed method, a cell suspension of islet pancreatic tissue culture in the amount of ~ 3 million cells in the growth medium volume ~ 0.2-0.45 ml). The further process of interpenetration occurs spontaneously, due to the physicochemical properties of the injection composition of the biodegradable matrix, which provides, at the same time, adequate respiration, nutrition, external function of the biomaterial and its fixation at the injection site. The transplantation of the injection composition of xenogenic biomaterial and biodegradable matrix to the recipient is carried out by introducing the contents of the syringe into the subcutaneous tissue of the abdomen under local anesthesia. Injection is performed from top to bottom through an injection needle catheter with a diameter of 2.0-2.2 mm.

Preliminary sampling of donor material, cultivation, primary processing, preservation and transportation is carried out according to the generally accepted method: under aseptic conditions of the laminar flow of sterile air, the biomaterial (the tail of the pancreas) is placed in a Petri dish containing physiological saline. Then it is transferred to another Petri dish and also under conditions of a laminar flow of sterile air, it is crushed as much as possible using eye scissors, periodically washing in physiological saline. After removing saline by low-speed centrifugation (1 thousand rpm), the crushed tissue is placed in a solution devoid of Ca ⁺⁺ and Mg ⁺⁺ ions. Excess solution is maximally removed and centrifuged (1 thousand rpm). The procedure is repeated several times. Immediately prior to transplantation, the cell suspension is centrifuged (1 thousand rpm) and extempore is injected into a syringe containing a biodegradable matrix.

The following examples 1 and 2 of a series of clinical studies illustrate the effectiveness of the proposed method for the treatment of diabetes mellitus type I (insulin-dependent form) and type II (non-insulin-dependent form), respectively.

Example 1

The introduction into the subcutaneous tissue of the abdomen of four patients suffering from non-insulin-dependent diabetes mellitus, a primary culture of islet tissue (3.0 million cells in 0.45 ml of growth medium) of the pancreas of newborn rabbits in the injection composition of the biodegradable matrix Sphero® GEL (2.0 ml) in accordance with the claimed invention showed that the dynamics of glycemia is not stable and has a smooth kinetics. Within one day after the operation, against the background of ongoing drug therapy, a temporary decrease in the glucose content in the blood plasma of the recipients was recorded from 20 to 15.0 mmol / L in the morning and up to 13.2 mmol / L in the evening. Over the next ten days, the amplitude of the oscillations (morning-evening) did not change (~ 7.5 mmol / L), demonstrating a steady tendency to a decrease in hyperglycemia up to 2-3 months of observation. Moreover, glycemia in its general dynamics practically normalized with reduced doses of sugar-lowering therapy (~ 2-4 times). All patients were in outpatient monitoring and control. The condition is satisfactory. There are no complications during the procedure and in the postoperative period.

Example 2

A total of 41 transplants were performed to 29 patients with type II diabetes mellitus in the general course of treatment (of which 7 repeated operations). The distribution of patients in comparison groups is presented in table 1. The age of the patients ranged from 37 to 72 years. Average age: 54.8 ± 9.89 years. Duration of the disease: from a newly diagnosed disease to 15 years or more. Objective selection criteria for patients are presented in table 2.

Table 1 Group name Place of introduction The number of cells, 10 ⁶ Volume ml Number of patients the introduction of β-cells of the islet tissue of the pancreas subfascial to the rectus abdominis muscle 2.0 20,0 9 the introduction of β-cells of islet tissue of the pancreas in the matrix in the subcutaneous tissue of the abdomen 3.0 2.2 10 the introduction of culture of islet tissue of the pancreas according to the claimed method in the subcutaneous tissue of the abdomen 3.0 2.2 10

table 2 Nosological form of the disease Randomization criteria Reference values diabetes blood glucose> 6.0 mmol / ml 4.10-5.9 mmol / ml urine sugar> 3.0 nmol / l 1.77-2.93 nmol / L HbA1a + b + c (glycated Hb) 5.6-7.5% of total Hb

The chosen scheme for introducing the culture of β-cells of the pancreatic islet tissue of newborn rabbits (under the fascia of the rectus abdominis muscle) showed that the glucose content in the recipient's blood plasma is not stable and has wave-like kinetics. Within one day after the operation, against the background of ongoing drug therapy, there was a temporary, but significant (p <0.05) decrease in the glucose content in the recipient's blood plasma (to 6.0 ± 3.1 mmol / L). In the next four days, the oscillation amplitude (morning-evening) decreased significantly (morning ~ 8.25 ± 1.18 mmol / l, evening ~ 9.45 ± 2.12 mmol / l; against: morning ~ 16 ± 4.5 mmol / l; evening ~ 9.2 ± 4.3 mmol / l), showing some unreliable tendency to increase hyperglycemia up to 3 months of observation (15.2 ± 4.8 mmol / l). By the sixth month, the glucose content in the blood plasma significantly decreased (11.8 ± 4.2 mmol / L), and by the 7-8th month it again increased (14.8 ± 5.4 mmol / L), not differing, in fact, from the preoperative level. The condition of all patients did not undergo significant changes, although the subjective assessment was very often unreasonably optimistic.

The general kinetics of glycemia in the postoperative period and the introduction of a culture of β-cells of the pancreatic islet tissue of newborn rabbits (into the subcutaneous tissue of the abdomen) as part of the biodegradable matrix Sphero® GEL did not affect the overall kinetics of glycemia. The glucose content was also unstable and had a clear wave-like kinetics similar to previous results (p> 0.05).

The selected scheme for the introduction of the primary culture of the islet pancreatic tissue of newborn rabbits in a biodegradable matrix (into the subcutaneous tissue of the abdomen) in accordance with the claimed method showed that the glucose content in the blood plasma of recipients, as in previous groups of patients, tends to wave-like kinetics, but differs more resilience. During the first days after the operation, against the background of ongoing drug therapy, a temporary decrease in the glucose content in the recipient's plasma was recorded to (6.0 mmol / L in the morning and 11.4 ± 3.1 in the evening). Over the next four days, the oscillation amplitude (morning-evening) decreased (morning ~ 9.2 ± 2.3; evening ~ 7.8 ± 2.0; against: morning ~ 12.2 ± 2.5; evening ~ 7.8 ± 2.3), showing some unreliable tendency to decrease hyperglycemia up to six days of observation, as in the comparison groups. It is important to note that in patients of this group, starting from six days, the sequence of high and low numbers of glucose in blood plasma changed: if before the fifth day of observation, morning results were ~ 2 times higher than evening ones, then in the period of 6-7 days this difference ceased to be determined, from 8 days: morning indicators again became higher than the evening ones by ~ 1.2-2.2 times. A comparative analysis, however, showed that the general dynamics of glycemia in the post-transplant period is preferable when transplanting a primary tissue culture according to the claimed method. A total of 10 patients were operated on, 2 of them re-operated. Both were in outpatient monitoring and control. The condition is satisfactory. There are no complications during the procedure and in the postoperative period.

Summing up, the claimed method allows within 1-2 months after the operation to normalize the level of glycemia and begin to reduce the dose of sugar-lowering therapy until it is completely canceled by the 3-6th month after the operation. If necessary, the operation can be repeated, but no later than 3.5–4 months after the first operation. In cases of severe initial condition of patients, according to indications, the following operation can be prescribed, but not earlier than 6 months after the last transplantation.

Claims (5)

1. A method for the treatment of diabetes mellitus, comprising introducing into the subcutaneous tissue of the abdomen a recipient of an injection composition in the form of a tissue culture and a biodegradable gel-containing matrix, wherein the primary culture of the pancreatic islet tissue of the donor in the form of a cell suspension is used as a tissue culture. 2. The method according to claim 1, in which a person is used as a donor. 3. The method according to claim 1, in which an animal is used as a donor. 4. The method according to claim 3, in which the primary culture of embryonic islet tissue of the pancreas is used as a tissue culture. 5. The method according to claim 3, in which the primary culture of fetal islet tissue of the pancreas is used as a tissue culture.