RU2140276C1 - Method of treating hepatic insufficiency in patients with different- genesis liver diseases - Google Patents

Abstract

FIELD: hepatology. SUBSTANCE: invention is designed for use in reanimation departments and departments dealing with intensive therapy in cases of acute and chronic hepatic insufficiency. Patient orally receives xenogenic lyophilized encapsulated hepatocytes - 2.0 in each capsules. In cases of acute hepatic insufficiency, 2 capsules 3 times a day are administered 1.5 hr before meat during 10 days. In cases of chronic hepatic insufficiency, 2 capsules 2 times a day are administered during 14 days. EFFECT: simplified treatment and considerably enhanced effect. 3 cl, 2 tbl, 2 ex

Description

 The invention relates to medicine, namely to hepatology, and may find application in intensive care units and intensive care units in the treatment of acute and chronic liver failure.

 Closest to the invention is a method of treating liver failure in chronic liver diseases by oral administration into the gastrointestinal tract of a suspension of hepatocytes in a Hanks solution based on 2 ml per 100 g of patient’s body weight at a concentration of 2x10 cells in 1 ml 30-40 minutes before food intake every two days on the third until a stable remission occurs, and before and after taking a suspension of hepatocytes, 100 ml of mineral water is administered (Copyright certificate N 2063760, A 61 K 35/407).

The disadvantages of this method are;
1. The impossibility of long-term storage of suspension, since the functional activity of hepatocytes decreases, and therefore, the effectiveness of the treatment of liver failure.

 2. A suspension of hepatocytes, getting into the stomach, is subjected to enzymatic treatment and destruction, which reduces the functional activity of hepatocytes and, consequently, the effectiveness of treatment.

 3. Suspension of hepatocytes requires expensive means of storage, for example, medium 199, Hanks solution.

 4. The proposed daily dose of suspension of hepatocytes (2 ml per 100 g of mass at a concentration of 2x10 cells in 1 ml) cannot be calculated on the predictable therapeutic effect, since the acidity in the stomach can vary in different ranges: from low to high values. In this regard, to determine the number of preserved functional hepatocytes is not possible, therefore, the therapeutic dose of hepatocytes varies in different limits.

5. Oral suspension of hepatocytes may cause aesthetically unpleasant sensations in some patients. The need to take mineral water causes some uncertainty: a) mineral water in case of diseases of the gastrointestinal tract is selected accordingly; b) it is not clear why it is necessary to take mineral water, with a view to what effect (electrolyte content, pH of the solution, the presence of carbon dioxide?),
The objective of the invention is to increase the effectiveness of treatment and simplify the method.

 The problem is solved in that in the method of treating liver failure in liver diseases of various genesis by oral administration of functionally active hepatocytes, xenogenic lyophilized hepatocytes are used in capsule form of 0.2 g, which are administered 1-2 capsules 1.5 hours before meals 2-3 times a day for 10 days, and in case of acute liver failure, 2 capsules are administered 3 times a day for 10-15 days, and then 1 capsule 3 times a day for the next 10-15 days, and with chronic liver failure Precisely injected 2 capsules 2 times a day for 10 days, and then 1 capsule 2 times a day for 14 days.

 Xenogenic lyophilized hepatocytes with prolonged storage at room temperature up to 3 years retain their functional activity.

 The clinical effect is achieved due to the entry of functionally active hepatocytes in the chyme into the small intestine, whence the active factors of hepatocytes - growth factor, enzymes - due to pinocytosis in an unchanged form and active state, enter the liver through the circulatory and lymphatic systems. Active factors of hepatocytes exert their specific effect on the patient’s liver, which consists in activating regenerative processes in the affected liver, stimulating the synthesizing and detoxifying functions of the patient’s functionally active liver cells.

 To substantiate the treatment method and determine the therapeutic dose, experimental studies were conducted on clinically healthy rats (106 animals) and sick dogs of various ages and breeds (1512 animals) with an average weight of 10 kg.

 The experimental rats were divided into three groups.

 So, in the first group, 5 ml of a solution with an active substance content of 0.3 was orally administered to white rats to detect side effects (toxicity); 0.4; 0.6 and 1.0 g. As shown by the results of studies, the drug did not have a toxic effect on experimental animals even with the introduction of high concentrations. Experimental animals did not die during the observation period for 14 days.

 The general clinical condition of the animals, as well as the dynamics of hematological and biochemical studies during the observation period were satisfactory.

 In the second group of animals, a model of toxic hepatitis was obtained by injecting animals with carbon tetrachloride, previously diluted in sterile vegetable oil in a ratio of 1: 1 at a dose of 0.6 ml / kg orally for three days.

 A wide range of doses, from 0.06 to 0.50 mg / kg body weight, was used to determine the optimal therapeutic dose of lyophilized xenogenic hepatocytes. The criterion of the therapeutic dose was considered such an amount of the drug, with the introduction of which the mortality decreased, the clinical condition of the animals improved. The drug was administered once or twice for five days. The analysis of the results showed the dependence of the survival of experimental animals on the amount of the drug (dose), as well as the frequency of administration. So, with a double administration of the same dose of the drug over 5 days, the mortality rate decreased in all experimental groups by 1.3-1.7 times compared with the groups where the drug was administered once, and with an increase in the dose to 0.5 mg / kg body weight this indicator is reduced by 4-5 times. Based on the data obtained, it is possible to consider the optimal therapeutic dose of 0.20-0.50 mg / kg of body weight twice within 5 days. The revealed optimal therapeutic doses of the drugs were confirmed once again by the method of pharmacological screening when administered to animals with varying degrees of liver damage.

 Studies have made it possible to establish the feasibility of using the drug for toxic hepatitis in the indicated doses, which allows to reduce the death of animals by 2-5 times and increase their life expectancy.

 In the third group of experimental animals, the kinetics of orally administered lyophilized hepatocytes stained with trypan blue was determined. For this, euthanasia of animals was carried out at certain time intervals. They established the movement of the cell suspension during the day; some of these cells reach the large intestine.

 Data from the results of experimental studies in rats were confirmed in the treatment of dogs with acute and chronic liver failure.

 The criterion for evaluating the effectiveness of the drug was the clinical condition indicators, the positive dynamics of a decrease in the concentration of toxic substances in the blood and the activity of serum cytoplasmic enzymes. By calculation, the optimal therapeutic dose of the proposed drug was compiled - 0.2 g per capsule.

 The therapeutic effectiveness of the capsules of the drug with the active substance content of 0.2 g was determined. The drug was prescribed 1 capsule in the morning and evening before feeding the animal for five or ten days, depending on the dynamics of clinical and biochemical parameters. The treatment results correlated with the dynamics of biochemical parameters (table. 1 at the end of the description). Table 1 presents the results of biochemical blood tests in the treatment of liver failure in the experiment.

 As can be seen from the table, in the process of treatment there is an improvement in clinical and hematological parameters: stabilization of blood pressure, decreased heart rate, respiratory rate; normalization of the level of proteinemia, glucoseemia and cholesterolemia; decrease in the concentration of toxic substances, the activity of certain liver enzymes, etc.

 The results formed the basis for oral administration of lyophilized hepatocytes in clinical practice.

 The method is as follows: in acute liver failure, the patient orally takes lyophilized hepatocytes containing 0.2 g in capsule form 2 capsules three times a day 1.5 hours before meals for 10-15 days, then 1 capsule three times a day for the next 10-15 days; in chronic liver failure in the decompensation stage, patients take 2 capsules orally twice a day 1.5 hours before meals for 10 days, then 1 capsule twice a day for 14 days.

Clinical example
Patient Ogorodnaya I.V., 35, medical history N 11525, was received on 08/02/97, was discharged on 08/26/97. Diagnosis: Overdose of paracetamol, acute hepatic-renal failure.

 Upon admission, the condition is serious, conscious, inhibited, worried by severe weakness, headache. On examination: marked yellowness of the skin, hemorrhage on the skin of the body, injection of sclera is noted. In the lungs, hard breathing, no wheezing. Hemodynamics is stable, heart rate - 78 beats / min, blood pressure -120/80 mm Hg The abdomen is swollen, peristalsis is sluggish, soreness in the right hypochondrium. Diuresis 50 ml per day. From the anamnesis it is known: on the eve of July 28, 1997, for the purpose of self-medication, I took 5 tablets of paracetamol during the day. 07.29.97, the patient appeared nausea, vomiting. 30-31.07.97 g noted yellowness of the skin, icteric sclera, decreased diuresis. Hyperfermentemia was noted in blood tests: AlAT - 524.5 n / mol s. l (30-40), AcAT - 110.5 n / mol s. l (30-40), LDH - 15.21 μmol / l (8.6), hyperbilirubinemia: total bilirubin - 208.67 μmol / l (5.0-30.0), direct - 154.0 μmol / l (2.0-26.0), prothrombin - 79% (80-95), hyperazotemia: urea - 14.6 mmol / L (10.0-12.0), creatinine - 354.0 μmol / L (80 , 0-120.0).

Ultrasound of the abdominal cavity: echographic signs of diffuse changes in the liver, parenchyma consolidation, enlarged spleen, portal vein - 1.5 cm, R _i (resistance index) - on the branches of the hepatic artery - 0.65, toxic changes in the kidneys, size 11x5.6 cm , R _i - 0.75. In the abdominal cavity, pleural cavities, a significant amount of free fluid.

 Complex therapy was carried out; active detoxification methods - hemodiafiltration (N5), plasmapheresis (N3), "roulette" dialysis (N2), vasculature and catheterization of the umbilical vein (BPV), followed by v / portal administration of glucose with B vitamins, lipoic acid, antispasmodics, reagents - Trental, chimes.

 Conservative therapy included: infusion therapy with forcing diuresis, correctly anemia, hypoproteinemia, water-electrolyte balance. Due to persistent hyperbilirubinemia (total bilirubin - 112.0 μmol / l), hyperfermentemia (AlAT - 107.8 n / mol s.L, AsAT - 121.0 n / mol s.L, LDH - 8.0 μmol / l), the course of protective hepatic therapy included oral administration of lyophilized hepatocytes for 10 days, 2 capsules 3 times a day, and then 1 capsule 3 times a day for the next 10 days.

After the treatment, a decrease in the concentration of total bilirubin to 87.3 μmol / L was noted in blood tests, the level of alanine and aspartic transaminases returned to normal. Restoration of diuresis was noted. In a control study of the abdominal cavity (ultrasound + Doppler), restoration of intrahepatic hemodynamics was noted; the diameter of the portal vein was 1.3 cm, R _i was normalized (0.58), and renal hemodynamics was restored. Moderate effects of portal hypertension persisted: an increase of up to 11x5 cm. After the treatment, the patient's condition improved: signs of encephalopathy disappeared, appetite appeared, and became more active. The concentration of bilirubin and enzymes reached normal values. The patient was discharged after 26 days.

Clinical example 2
Patient Fokin V.A., 49 years old, medical history 4508, was admitted March 26, 1997. Diagnosis: Cirrhosis of the liver, decompensation. Portal hypertension: hepatosplenomegaly, varicose veins of the esophagus, ascites. Complaints at admission: weakness, sleep disturbance, headache. From the anamnesis it is known that the patient suffers from cirrhosis of the liver for 3 years, abuse alcohol.

The last deterioration three weeks ago - there was weakness, decreased appetite, disturbed sleep, appeared inadequate behavior. The abdomen enlarged, swelling appeared on the lower extremities. On examination: skin, icteric sclera, liver on palpation +6 cm from under the edge of the costal arch, painful. Instrumental studies were conducted. Liver scintigraphy: irregularly shaped liver, fuzzy, fuzzy contours, dimensions 13x9x20 cm, distribution of radiopharmaceuticals diffusely uneven, with a decrease in radiopharmaceutical in the right lobe, spleen 16x12 ohm, radiopharmaceutical capture 48%. Signs of cirrhosis, splenomegaly, portal hypertension. Ultrasound revealed an increase in the liver, compaction of the parenchyma, the intrahepatic network was expanded due to the portal system, the diameter of the portal vein was 2.2 cm, R _i was 0.80, and the diameter of the splenic vein was 1.6 cm. Free fluid was present in the abdominal cavity. In admission to blood tests, hyperbilirubinemia is noted: total bilirubin -184.3 μmol / L, direct - 137.0 μmol / L, cholesterol - 1.8 mmol / L, beta-lipoproteins - 1.7 g / L, hypokalemia - 3.5 mmol / l, hypoproteinemia: total protein - 58.0 g / l, hyperfermentemia: AsAT - 72.5 n / mol s.L, LDH - 10.86 μmol / s. L, GTP - 99.9 IU / L, prothrombin - 85.0%.

 The treatment was carried out: infusion therapy with forced diuresis - glucose, B vitamins, antispasmodics, antibiotics, diuretics. On the 5th day from the day of admission, the patient received orally 1.5 hours before meals lyophilized hepatocytes 2 capsules twice a day for 10 days, then 1 capsule 2 times a day for 14 days. On the 7th day of taking lyophilized hepatocytes, a decrease in the concentration of total bilirubin was noted by 45%, the lipid concentration increased: cholesterol - 3.3 mmol / l, beta-lipoproteins - 3.0 g / l. The concentration of total protein was 63.0 g / L. The concentration of AsAT, LDH normalized on the 14th day of hepatocyte intake.

 These data are confirmed by ultrasound: signs of portal hypertension have decreased, free fluid - a minimal amount. The patient became active, adequate, improved appetite, normalized sleep. The patient was discharged on the 34th day of the disease with subsequent remission of 6 months.

 According to the proposed method, 47 patients were treated. In all patients, the course of the disease corresponded to hepatopathy I - II degree. In the treatment of this group of patients used lyophilized hepatocytes in capsule form in the form of oral administration.

 The dynamics of changes in blood biochemical parameters during treatment are presented in table. 2 (see the end of the description). As can be seen from the table, a 60% decrease in bilirubinemia was noted on the 6th day, and on the 14th day, the concentration of total bilirubin and its fractions decreased by 80%.

 Fermentemia decreased on the 14th day: AsAT - by 34%, AlAT - by 60%. By the end of the third week of treatment, blood biochemical parameters returned to normal. During the treatment, the patients' well-being was noted, the manifestations of encephalopathy decreased: sleep normalized and appetite appeared. In patients with chronic liver diseases, signs of portal hypertension decreased, which was confirmed by instrumental studies (ultrasound, liver scintigraphy).

 Thus, the method of treating patients with acute and chronic liver diseases with lyophilized hepatocytes can be considered as more effective, simplified and cheapened compared with the suspension of live hepatocytes, the advantage of this method of treatment is high functional activity, dosage accuracy, preservation of the drug from the damaging effects of gastric juice , the duration of their storage (for 3 years), which allows the treatment of patients with various degrees of severity of dysfunction Yeni.

Claims (3)

 1. A method for the treatment of liver failure in liver disease of various origins by oral administration of functionally active hepatocytes, characterized in that xenogenic lyophilized hepatocytes are used in capsule form of 0.2 g, which are administered 1 to 2 capsules 1.5 hours before meals 2 - 3 times a day for 10 to 30 days.  2. The method according to claim 1, characterized in that in acute liver failure, 2 capsules are administered 3 times a day for 10 to 15 days.  3. The method according to claim 2, characterized in that in chronic liver failure, 2 capsules are administered 2 times a day for 10 days, and then 1 capsule 2 times for 14 days.

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