RU2248796C2 - Pharmaceutical composition (variants) no containing proteins and peptides for neutralization and/or removing endotoxins from body - Google Patents

Abstract

FIELD: medicine, toxicology, pharmacy.

SUBSTANCE: according with the first variant the composition contains neutral lipid and therapeutically effective amount of cholanic acid or cholanic acid salt and phospholipid. Neutral lipid presents in the amount from 3% to 50% by mass relatively to the total amount of lipid. According with the second variant the composition contains from 3% to 30% by mass of bile acid or bile acid salt, from 3% to 50% by mass of neutral lipid and from 10% to 95% by mass of phospholipid. Composition is designated for treatment in poisoning with endotoxins. Composition no containing peptides and proteins but containing the combination of phospholipid with cholanic acid proves effective relief or prophylaxis of endotoxemia.

EFFECT: enhanced effectiveness and valuable medicinal properties of composition.

23 cl, 10 dwg, 2 tbl, 10 ex

Description

Technical field

The invention relates to the treatment of endotoxemia caused by endotoxins, in particular to the treatment of endotoxin poisoning by administering various formulations into the body that neutralize and / or remove endotoxins from the body, as well as for prophylaxis using these formulations.

State of the art

Endotoxin bacterial toxic shock is a condition, often fatal, triggered by the release of lipopolysaccharide (LPS) from the outer membrane of most gram-negative bacteria (for example, Escherichia coli: Salmonella tymphimurium). The structure of bacterial LPS is fairly well explained, and the only molecule called lipid A is linked to the acyl chains through its glucosamine chain (see Paetz, Ann. Pev. Biochem. 59: 129-170 (1990)).

The lipid A molecule serves as a membrane retainer of the structure of lipopolysaccharide (LPS), which is involved in the development of endotoxin shock. It should be noted that LPS molecules are characterized by a lipid A type structure and part of a polysaccharide. This part may differ in details in various LPS molecules, but it retains the general structural basis of endotoxins. It would be wrong to say that the LPS molecule is the same in all bacteria (see Raetz, above). Usually, experts call various LPS molecules “endotoxins”, and this term will be used hereinafter to refer to LPS molecules.

US Pat. No. 5,128,318, the description of which is incorporated by reference, indicates that reconstituted particles containing both the apolipoprotein associated with HDL and a lipid capable of binding endotoxin to inactivate it can be used as effective materials to attenuate toxicity caused by endotoxins.

In previous applications, which are used in the present description by reference, it was indicated that other materials can be used to suppress toxicity caused by endotoxins. In particular, it was found that apolipoproteins are not required in the reconstructed particles and that the reconstructed particle may contain a peptide and a lipid, where the peptide is not an apolipoprotein.

The inventors have also found that toxicity caused by endotoxins can be controlled by sequential administration of an apolipoprotein or peptide, and then a lipid. Serially introduced components form a reconstructed particle and become active in the elimination of endotoxins.

It has also been found that at least some patients have natural apolipoprotein levels that are higher than normal and allow for effective therapy of endotoxemia by administering reconstituted particles not containing apolipoprotein or peptide but containing the lipid described.

In addition, the invention disclosed in these applications involves the use of reconstructed particles and the aforementioned components for the prevention of toxicity caused by endotoxins by introducing a prophylactically effective amount to prophylactic subjects. Such subjects include infected patients or recovering from surgery. These patients sometimes have very low plasma HDL levels, decreasing to 20% of normal levels. In these cases, early prophylaxis using HDL to compensate for these decreases is desirable.

It has been unexpectedly discovered that phospholipids can be used alone or in combination with additional materials, such as neutral lipids, cholates, and the like, as an effective means to weaken and / or prevent endotoxemia. In particular, it is preferred to use phosphatidylcholines (hereinafter referred to as PF) both individually and in combination with other phospholipids, such as sphingolipids, in compositions that are substantially free of peptides and proteins, for example apolipoproteins or peptides derived from them. Neutral lipids, such as mono-, di-, and triglycerides, can be mixed with phospholipids until the total amount of natural lipids is below a certain mass percentage when using these formulations in the form of balls for intravenous administration. With other methods of administration, for example intravenously by continuous infusion, the percentage by weight is not so important, but desirable.

Particularly preferred embodiments of the invention involve the use of emulsions in which bile acid or its salt is used together with a phospholipid or a neutral lipid.

The effectiveness of bile acids and their salts - cholates in the treatment of endotoxemia is shown. These bile acids may be used alone or in combination with one or more phospholipids and / or a neutral lipid, such as phosphatidylcholine and / or triglyceride.

The invention is described in more detail below.

A brief description of the illustrations.

Figures 1 and 2 show the results obtained by testing various formulations in a model where the neutralization of endotoxin was determined by detecting the release of TNF into a model of human whole blood. Figure 1 shows the value of the protein, and figure 2 - phospholipid. The tested formulations included natural lipoproteins (VLDL, LDL, HDL), reconstructed HDL (R-HDL) and INTPALIPID ® formulations, as well as emulsions containing phospholipid and protein.

Figures 3 and 4 show the comparative value of triglyceride (neutral lipid) and phosphatidylcholine, phospholipid in the same model.

Figure 5 provides toxicity information for the introduction of various PF and PF / TG compositions into a mouse model using a model infected with E. coli LPS. with a fatal outcome of 55%.

Figure 6 shows data comparable to the data obtained above when testing a model of whole human blood, but when using a phospholipid with unesterified cholesterol, sphingomyelin or a mixture of the latter instead of triglycerides.

Figures 7 and 8 show results comparable to those shown in figures 1 and 2, in which case a phospholipid, unesterified cholesterol and / or sphingomyelin are mixed with triglycerides or esterified cholesterol as a neutral lipid.

Figure 9 presents a comparison of the results obtained from the introduction of emulsions containing cholesterol ester and triglyceride in an in vivo mouse model.

Figure 10 shows a diagram of the theoretical amount of triglycerides released into the blood after the introduction of various TG-containing formulations, with toxicity thresholds. The abbreviation "OPP" means "general parenteral nutrition", and "SI" means the compositions according to the invention.

Detailed Description of Embodiments

Example 1

Factors that influence the stimulation of TNF-α by LPS while maintaining the integrity of the interaction of plasma proteins and blood cell elements can be adequately investigated in vitro on the whole human blood system. Such a system was used to determine which of the lipoprotein components are important in neutralizing LPS.

The following materials were tested:

reconstructed high-density lipoprotein (R-HDL), natural lipoproteins (VLDL, LDL, HDL) of blood plasma, serum with a lack of lipoproteins (SNLP) and triglyceride-rich emulsion 20% INTPALIPID ® (a mixture of triglycerides and phospholipids).

Blood was collected in a heparinized tube, diluted with Hank's balanced salt solution (hereinafter CCPX), or the test material dissolved in CCPX was poured. The resulting material was poured into Starstedt tubes (250 μl / tube). LPS was dissolved in pyrogen-free saline containing 10 mM HEPES buffer and added (2.5 μl) to a final concentration of 10 ng / ml. After incubation for 4 hours at 37 ° C, the tubes were cooled to 4 ° C, after which they were centrifuged at 10,000 × g for 5 minutes. The supernatant was discharged and TNF-α was determined using a known enzyme immunosorbent assay (ELISA).

Table 1 shows comparative data on the compositions of the tested materials. Figure 1 and 2 presents the results. The data are presented in the form of graphs of the dependence of the amount of TNF-α produced on the concentration of added protein (Fig. 1) and phospholipid (Fig. 2). To show the wide range of concentrations used, a logarithmic scale was used, where 10 ° corresponds to 1 mg / ml. All incubated whole blood systems contained 10 ng / ml E.coli 0111: B4 LPS supplemented with one of the formulations, as an explication to figures 1 and 2.

The fact that the effectiveness of the materials is different when plotting the protein content (FIG. 1) and is similar when applying the phospholipid content (FIG. 2) indicates that the phospholipid is an important component. This was also confirmed by the fact that a protein-free lipid emulsion is more effective than natural HDL, but less effective than R-HDL. Protein is not so important for neutralization.

Table 1

Composition of natural lipoproteins and reconstructed HDL

Lipoprotein CT TG FH Protein Class Density (g / ml) Weight (%) VLDL <1.006 22 53 eighteen 7 LDL 1.007-1.063 48 eleven 22 20.9 HDL 1,063-1,21 eighteen 8 22 52 R-HDL 1,063-1,21 - - 79 21 SNLP <1.21 0 0 2 98 Intralipid - 1 93 6 0

Example 2

In this example, protein-free lipid emulsions containing different amounts of neutral lipid were tested on whole human blood. In this case, the same analysis was used as in the in vitro study of whole human blood in Example 1.

All particles described herein were obtained according to the same procedure, which involves mixing a phospholipid, sphingomyelin or phosphatidylcholine, triolein and / or ester of unesterified cholesterol dissolved in chloroform, and weighing them before placing in a flask. Vitamin E (0.02% w / v) was added as an antioxidant. Then, a dry lipid film was prepared by blowing nitrogen gas or argon over the sample. Then pyrogen-free saline was added to the flask, and then the contents of the flask were mixed in a vortex mixer until all of the lipid was suspended. The solution was then homogenized in a high pressure homogenizer. Samples containing phosphatidylcholine (FC), with or without triolein, were cyclically passed through the homogenizer 10 times at a pressure of 137895 kPa (20,000 psi). Samples containing cholesterol ester with one or more other lipids were cyclically passed 15-20 times at a pressure of 206842 kPa (30,000 psi). The homogenized solutions were filtered using a syringe with a filter nozzle having pores of 0.45 μm, the filtrate was left at room temperature (for 3 days) until use. Figures 3 and 4 show the results of these studies, where the data on the production of LPS-dependent TNF-α depend on the concentration of added triglyceride (Fig. 3) or phospholipid (Fig. 4). The compositions, as shown in the explication, contained 7% triglyceride (TG), 45% TG, 89% TG, 94% TG, P-HDL or phospholipid without TG (shown only in FIG. 4). A composition with 89% TG contains 10% INTPALIPID ®, and 94% TG contains 20% INTPALIPID. In all other experiments, egg phosphatidylcholine (FC) and triolein were used.

These results show that protein-free formulations vary greatly in their effect of triglyceride content, but they are very similar when studying the effects of phospholipid (PC) content. This confirms the role of phospholipid, since only phospholipid is effective, although to a lesser extent than emulsions containing up to 45% TG.

Example 3

Next, the experiments were continued in vivo on a mouse model that was chosen as a reliable system for predicting the effectiveness of actions on humans.

In these experiments, the mice were injected, in the form of balls, a sufficient number of compounds described in example 2, as well as other compounds (pure phosphatidylcholine, 7% TG, 25% TG, 45% TG, 71% TG, 81% TG, 94% TG ) or a control saline solution, with doses of phospholipid (200 mg / kg or 400 mg / kg), together with 25 mg / kg E. coli 0111: B4 LPS. The control group received intravenous saline in a volume corresponding to the volume of the emulsion. Survival after 72 hours is shown in FIG. 5. Of the 344 animals in the control group, 155 survived.

The use of FC alone gave a moderate protective effect, statistically insignificant at a confidence level of 95%, while formulations containing 7%, 45%, and 71% TG significantly increased survival. Compositions containing 80% and 89% TG provided a minimal effect, while survival rates containing 94% TG decreased.

When the doses were increased to 400 mg / kg PF, emulsions containing 89% and 94% TG significantly reduced the lifetime, probably due to the toxic effect of TG, which is explained below.

Example 4

In examples 1-3, it was found that phospholipids are an active means of suppressing endotoxemia. The fact that non-polar lipids, instead of triglycerides, can form emulsions with phospholipids, and not with PF, suggests that other substances can be used, for example, sphingomyelin (another phospholipid), unesterified cholesterol (polar neutral lipid), and mixtures thereof. Etherified cholesterol (non-polar ether), squalene (hydrocarbon) and vitamin E (non-polar antioxidant) can also be used. To test these substances, a series of experiments were carried out, with the analysis of the model of whole human blood in example 1 and the survival of mice in example 3.

Emulsions were prepared, as described above, from pure phosphatidylcholine, phosphatidylcholine with 10% (by weight) unesterified cholesterol, 10% (by weight) sphingomyelin, or only 10% of a mixture of unesterified cholesterol and sphingomyelin. Emulsions were added to whole blood at a concentration of 100 mg / dl in relation to PF and 10 ng / ml LPS. The mixture was incubated and the isolated TNF-α was measured.

The experimental results are shown in Fig.6. The isolation of TNF-α was significantly reduced when using only PF. Emulsions containing unesterified cholesterol, sphingomyelin, or a mixture thereof, also inhibited the release of TNF-α.

Example 5

Whole blood analysis was also used to determine the effect of unesterified cholesterol and / or sphingomyelin on emulsions containing a neutral lipid. In addition, emulsions at 100 mg / dl PF were used. The various compositions (in mass terms) are presented in table 2.

table 2

Emulsion Composition FC with 45% TG 55:45 ФХ + ТГ + С 54.4: 45.3: 0.3 FH + TG + SF 51.6: 43.3: 5.4 ФХ + ТГ + С + СФ 51.4: 42.9: 0.3: 5.4 FH + EH 54.5: 45.5 ФХ + ЭХ + С 54.4: 45.3: 0.3 FH + EH + SF 51.6: 43.0: 5.4 ФХ + ЭХ + С + СФ 51.5: 42.9: 0.3: 5.4

Figures 7 and 8 show the results of the experiment. PF emulsions prepared with a neutral lipid in the presence or absence of polar lipids showed inhibitory ability. In addition, the LPS concentration was 10 ng / ml, i.e. clinically relevant endotoxin concentration. Emulsions containing cholesterol ester were less effective than emulsions containing TG, while emulsions containing unesterified cholesterol did not suppress TNF-α, as did emulsions that did not contain cholesterol. The addition of sphingomyelin in the emulsion led to an increase in suppression of the production of TNF-α.

Example 6

Emulsions containing cholesterol ester were tested in vivo in the model used in Example 3 with a lethal dose of endotoxins. Emulsions were prepared from PF and TG or PF and cholesterol ester (SEC), injected in a single dose of 200 mg / kg PF in the form of a ball together with 25 mg / kg E.coli 0111: B4 LPS (lethal dose) into the tail vein. The control group received intravenous saline in a volume corresponding to the volume of the emulsion.

Figure 9 gives comparative results of the action of emulsions containing PF and TG. One and the other emulsions were tested in at least two experiments using 16 or more animals.

As can be seen in Fig.9, emulsions containing 7% or 45% SEC by weight, significantly increased survival. These results, considered in conjunction with the results in Example 5, show that EC can be a substitute for TG when creating emulsions that neutralize endotoxins.

Example 7

Protein-free emulsions of phospholipid with triglyceride effectively block the production of TNF-α in whole blood, activated by LPS. Theoretically, these emulsions can also be effective on living objects if they can be safely administered in doses that provide protective plasma phospholipid concentrations. Our previous experiments with R-HDL suggest that the minimum dose of phospholipid is approximately 200 mg / kg. Using this dose and plasma volume of 4.5% of body weight, it is possible to calculate the concentration of triglyceride expected in the plasma after the introduction of emulsions with a successively increasing content of triglyceride. The result is shown in figure 10 in the form of a smooth line, bending upward with increasing TG content (% by weight). Plasma TG concentrations rarely exceed 1000 mg / dL in healthy adults, even after ingestion of fatty foods. According to clinical data, pancreatitis occurs in patients with a plasma TG concentration above 2000 mg / dl (Farmer et al., Amer. J. Med. 54: 161-164 (1973); Krauses et al., Amer. J.

Med. 62: 144-149 (1977); Glueck et al., J. lab. Clin. Med. 123: 59-61). Plasma TG concentrations above 4000 mg / dl are extremely rare and cause serious concern. 10, the last two thresholds are shown by horizontal lines. With the introduction of 10% or 20% INTPALIPID ® to obtain a dose containing 200 mg / kg phospholipid, a significant increase in plasma TG concentrations is expected (see two contour circles) beyond safe limits. In contrast, the introduction of emulsions containing 7%, 45%, 71% or 78% (solid squares from left to right) increases the concentration of TG in the plasma to 136, 477, 1300 or 2000 mg / dl, respectively. Emulsions with a TG content of up to 50% are not expected to be toxic due to the presence of TG.

Example 8

The effectiveness of combinations of phospholipid and bile acid, i.e. sodium cholate was tested in an experiment similar to that described in the previous examples. However, the method by which the compositions were introduced, introduced for testing on animals, had a difference. In this and subsequent examples, compositions were prepared using a high pressure homogenizer

"Microfluidizer". This unit makes scaling easy.

Liquid triolein or liquid soya triglyceride was dissolved in an appropriate mass of water or water with 9 mM, 18 mM or 36 mM sodium cholate. Solid granular phosphatidylcholine was weighed on paper, and then slowly added to the solution with stirring. This takes about 3-5 minutes to disperse the lipid. After dispersion, the resulting material was poured into a high pressure homogenizer. This device uses hydraulic pressure to turn on a pump, which in turn directs two opposing jets of samples. Pressure can reach 172.4 kPa (25,000 psi). Upon impact, the jets under pressure pass into the cross-shaped hole, thereby homogenizing the sample.

The sample was passed through the homogenizer several times, with one pass being the time required to pump the entire sample through the device. Samples were passed 20 times to achieve the required level of homogenization. Dextrose was added to a final 5% concentration.

Endotoxin purified from E. coli 0111: B4 (40 mg / kg) and the emulsion described below (200 mg phosphatidylcholine / kg) were mixed at room temperature and immediately introduced into C57BL6 / J mice (weight 19-30 g) by intravenous injection into the tail vein. Those mice that received only cholate were given a volumetric dose of sodium cholate equal to the dose of cholate / EML (emulsion) at the same cholate concentration. The control mice were injected with the same dose of 5% dextrose solution to ensure plasma osmolality.

The experimental results are shown in table 3. In this case, an emulsion containing phosphatidylcholine and 7% triglyceride described in the previous examples was used. Sodium cholate was added at the indicated concentration to the starting materials until an emulsion was obtained.

Table 3

Protecting mice from a lethal dose of endotoxin

Time watch) The control 7% TG 7% TG + HC 18 MMHK 9 mm 18 mm 36 mm lack of PF or TG + FH The number of surviving mice (N) 0 28 28 8 16 8 8 8 24 nine 12 4 fifteen 8 8 8 48 5 10 2 fifteen 8 8 8 72 2 5 1 fifteen 8 8 8 96 1 0 1 fifteen 8 7 8

The content of emulsions by weight was as follows. When using 9 mM cholate, the percentage by weight in the emulsion was: 7% cholate, 6.1% triglyceride and 86.9% phosphatidylcholine; at 18 mM cholate: 13.1% cholate, 5.7% triglyceride and 81.2% phosphatidylcholine; at 36 mM cholate: 23.2% cholate, 5% triglyceride and 71.8% phosphatidylcholine.

It should be noted that the amount of LPS administered in these experiments (40 mg / kg) is significantly higher than the amount used in the mortality study in the previous examples. The purpose of administering these increased doses was to suppress any protective effect of phosphatidylcholine and / or triglyceride. Thus, it follows from these experiments that the bile salt, i.e. sodium cholate, have a protective property.

Experiments with other salts of bile acid and taurine containing bile salts are not described herein. Among the bile acids, it should be noted, for example, allodeoxycholic, lithocholic, hyodeoxycholic, hyocholic acids, α-β and ω-muricholeic acids, murodeoxycholic, ursodeoxycholic, ursocholic acids and all their salts, such as sodium salts or taurine, or conjugated glycides. See Hoffmann below.

Example 9

Next, experiments were conducted, in the first of which the survival of mice used as experimental animals was investigated. Moreover, they were divided into 4 groups. The first group, the control, was injected with a 5% dextrose solution. The second was an emulsion containing 93% (by weight) phosphatidylcholine and 7% (by weight) triglyceride and prepared as described above. This emulsion contained 5% dextrose and soy phospholipids with approximately 50 mg / ml lipid. An emulsion similar to the emulsion introduced to the second group was added to the third and fourth groups of animals with the addition of 18 mM sodium cholate or 18 mM sodium deoxycholate. In this experiment, the same procedure was used as in Example 8. Survival was determined 72 hours after infection; the data are summarized in table 4.

Table 4
The effect of adding bile acid to a 7% triglyceride emulsion on mouse survival after 72 hours Group No. Survival rate (%) Value
p / group 1 2 3 1 5% dextrose 28 4 - - - 2 7% TG 64 8 NS * - - 3 Sodium Cholate + 7% TG 16 94 0.00001 0.00001 - 4 Sodium Dioxicholate + 7% TG 8 75 0.0001 0.00001 NS * NS - unacceptable

Note that the statistical significance of animal survival comparisons was verified using the Wilcox method using a computer program. Comparative data with control group 1 are placed under the index "1", a comparison with group 2, the animals of which were given a 7% emulsion - "2", and a comparison with group 3 of animals, which were injected with an emulsion with sodium cholate - "3".

Both survival (%) and statistical analysis reflect a definite, unexpected advantage in formulations containing bile salts.

Example 10

The second group of experiments was tested on a rabbit model. In this model, the isolation of TNF-α (necrosis factor of neoplasms) was determined.

The rabbits were divided into 3 groups and a 5% dextrose solution, an emulsion of phospholipid and triglyceride (93% / 7%) described above, or the same emulsion, but with 18 mM cholic acid, were introduced. All emulsions were corrected for a 5% dextrose solution, as in Example 9. The rabbits were previously injected with an emulsion ball and after 2 hours, 100 μg of E.coli 0111: B4 LPS. In addition to the emulsion ball, a composition of 50 mg lipid per kilogram of body weight per hour was continuously infused intravenously. Intravenous infusion was carried out for 3 hours after challenge.

Blood was drawn from rabbits in the area of the line separating the arch and base of the skull, 30 minutes after the preliminary injection of the ball and every hour for 5 hours after administration.

Table 5 presents the maximum values of TNF-α. They are registered 2 hours after administration of endotoxin.

The statistical significance of the indicators was determined using the well-known Student criterion. As can be seen from table 5, the indices of TNF-α decreased significantly after the introduction of 18 mm cholic acid.

Table 5
The effect of emulsion on the production of TNF-α in rabbits Emulsion TNF-α Relevance (ng / ml) N R 5% dextrose, control 134 ± 70 nine 7% TG emulsion 68 ± 5 5 <0.05 7% TG emulsion + 18 mM cholic acid 39 ± 20 4 <0.01

These examples disclose in detail an invention that provides, for one goal, the reduction or prevention of endotoxemia by administering to the patient an effective amount of phospholipid with which endotoxins are associated. Associations of phospholipid and endotoxins are then removed from the patient's body as a result of normal biological processes, well known to those familiar with lipoprotein particle removal processes. Phospholipid inhibits endotoxin in association with it.

It can be seen from these examples that the administration of any of the cholanic acids or salts of cholanic acids, such as bile acid or a salt thereof, can also be used instead of phospholipids to achieve the same effect, i.e. weakening or preventing endotoxemia. Thus, formulations not containing peptides and proteins, but containing only bile acid or a salt of bile acid, or both, as well as a phospholipid, can be used to treat endotoxemia. A description of cholanic acids (see, for example, Hofmann, Hepatology 4 (S): 4S-14S (1984)) is hereby incorporated by reference, in particular, page 5S, FIGS. 1 and 2, which show characteristic structures of cholanic acids .

The invention can be used preferably for the treatment of people, but its use in veterinary medicine is not ruled out.

The term “attenuation”, as used herein, means a treatment to reduce the severity of endotoxemia caused by any of the endotoxins produced, for example, by gram-negative bacteria (S. tymphimirium, E. coli, etc.). Prevention can be carried out by the introduction of an appropriate agent at the moment when the danger of endotoxin poisoning arose or is assumed. A classic example is surgery. So, the patient, who is preparing for surgery, you can enter the active ingredient.

The effective amount of phospholipid and bile acid needed to treat a patient may vary. Typically, a dose with a total weight of about 200-800 mg of phospholipid per kilogram of body weight is preferred, although this amount may be more or less depending on the severity of endotoxemia or the degree of risk for prevention. The used dose of cholanic acids and salts, for example bile acids and their salts, contains about 10-300 mg per kilogram of body weight, more preferably about 275 mg per kilogram of body weight.

It is desirable to administer bile acid / its salt and phospholipids with neutral lipids, but this is not necessary, since an emulsion of phospholipids containing no neutral lipids is also provided. The advisability of combined administration of phospholipids is due to the fact that neutral lipids and phospholipids are combined into particles that are similar to lipoproteins, but differ from them in that they do not contain protein peptide components, which of course are always present in lipoproteins.

Especially desirable are those forms of treatment where the phospholipid is phosphatidylcholine, for example egg yolk phosphatidylcholine, soybean phosphatidylcholine or sphingolipid. As bile acid / its salt, cholic acid and / or its salts are preferred, such as sodium cholate, sodium deoxycholate and sodium chenodeoxycholate. Regarding neutral lipids, it is preferable to use cholesterol ester or triglyceride, but other neutral lipids, such as squalene or other hydrocarbon oils, di- and monoglycerides and antioxidants such as vitamin E, can also be used.

Forms for administration may be different, with spherical or others for intravenous administration are particularly preferred. When using a spherical form, for example, with triglyceride, caution should be exercised when prescribing a dose. It is well known that triglycerides are toxic if they are introduced in too much. But a specialist can easily choose such a composition to eliminate or reduce the risk of triglyceride poisoning. Typically, when using a spherical shape, the mass fraction of triglyceride or other neutral lipid should be approximately not more than 80%, preferably not more than 70%. More preferably, the mass fraction of neutral lipid in the ball should be about no more than 50%.

However, when non-spherical forms are used, for example, other forms for intravenous administration, the risk of poisoning is reduced. However, the limits described above are preferred for intravenous or other routes of administration, although it is obvious that they are not required. Doses of bile acids and their salts are preferably from about 25 to 500 mg / kg body weight or more, preferably from about 50 to 100 mg / kg body weight. Preferred doses of phospholipids are preferably 100 to 1000 mg / kg body weight. The total doses are given here, however, they will vary depending on the patient and route of administration.

As indicated above, in compositions containing no proteins and peptides, at least one phospholipid or bile acid / salt thereof is required. Of the phospholipids, the presence of at least one neutral lipid, for example a triglyceride, diglyceride or monoglyceride, is preferred. These compositions may include additional materials, such as sterols (e.g., cholesterol, β-sitosterol), esterified or unesterified lipids (e.g., cholesterol ester or unesterified cholesterol), hydrocarbon oils, such as squalene, antioxidants, such as vitamin E, but they are not required. There is no doubt that any such formulation may include more than one phospholipid and / or more than one neutral lipid. When using a combination of a neutral lipid and a phospholipid, the neutral lipid should be present in an amount of about 3% to 50% by weight relative to the total amount of lipid in the composition.

Bile acid or its salts can be used alone or in combination with a phospholipid, a neutral lipid. As for additional materials (for example, phospholipids or neutral lipids), the species mentioned above are preferred.

In particular, the invention also provides compositions for treating endotoxemia. One example embodiment of the invention is a composition containing at least one of the bile acids / its salt, a phospholipid and a neutral lipid, which generally contains the active ingredient in an amount that attenuates the manifestation of endotoxemia. This composition preferably contains from about 5% to 30% by weight of bile acid / its salt, from about 3% to 50% by weight of neutral lipid and from about 10% to 95% by weight of phospholipid. Particularly preferred are formulations containing about 10-15% by weight of bile acid / salt thereof, from about 5% to 10% by weight of a neutral lipid, and the rest is a phospholipid.

It should be noted that these mass fractions relate to compositions containing three components. When combining a three-component system with, for example, a carrier, adjuvant, optional ingredients similar to those described above, the indicated mass fractions with respect to the whole composition will be lower. It should be borne in mind that in all cases, such therapeutic compositions do not contain proteins and peptides.

If the compositions not containing proteins and peptides also do not contain bile acid or its salt, then at least about 3% to about 50% by weight of the neutral lipid can be introduced into them, the rest is at least one phospholipid. A neutral lipid is predominantly triglyceride or any additional neutral lipids described above. In addition, phospholipid is predominantly phosphatidylcholine. Other aspects of the invention are clear to the skilled person and there is no need to re-list them here.

It is understood that the description and examples serve to illustrate the invention and do not limit the scope, and one skilled in the art can propose other embodiments of the invention that do not go beyond its essence and scope.

Claims (23)

1. Protein and peptide-free pharmaceutical composition suitable for neutralizing and / or removing endotoxins from the body by intravenous administration, containing a neutral lipid and a therapeutically effective amount of cholanic acid or a salt of cholanic acid and phospholipid, where the neutral lipid is present in an amount of from about 3 to approximately 50% of the total amount of lipid. 2. The pharmaceutical composition according to claim 1, wherein the cholanic acid or cholanic acid salt is bile acid or a bile acid salt. 3. The pharmaceutical composition according to claim 1 or 2, where the phospholipid is a phosphatidylcholine. 4. The pharmaceutical composition according to any one of claims 1 and 2, where the phospholipid is a sphingolipid. 5. The pharmaceutical composition according to claim 1, where the neutral lipid is a triglyceride. 6. The pharmaceutical composition according to claim 1, where the neutral lipid is an ester of cholesterol. 7. The pharmaceutical composition according to claim 2, where the bile salt is selected from sodium cholate, sodium deoxycholate and sodium chenodeoxycholate. 8. The pharmaceutical composition according to claim 7, where the bile salt is sodium cholate. 9. The pharmaceutical composition according to claim 1, containing phosphatidylcholine, triglyceride and sodium cholate. 10. The pharmaceutical composition according to claim 9, containing phosphatidylcholine and triglyceride in a mass ratio of 93: 7. 11. The pharmaceutical composition according to claim 9, where sodium cholate, phosphatidylcholine and triglyceride have a total mass ratio of 13.1: 81.2: 5.7, respectively. 12. Not containing proteins and peptides, a pharmaceutical composition suitable for neutralizing and / or removing endotoxins from the body, containing bile acid / bile salt, phospholipid and neutral lipid, said composition containing from about 5 to about 30% by weight of bile acid / bile salts, from about 3 to about 50% by weight of a neutral lipid and from about 10 to about 95% by weight of a phospholipid. 13. The pharmaceutical composition according to item 12, containing about 10-15% by weight of bile acid / bile salt, from about 5 to about 10% by weight of a neutral lipid and the rest is a phospholipid. 14. The pharmaceutical composition according to item 12 or 13, which is suitable for intravenous administration. 15. The pharmaceutical composition according to item 12, where the phospholipid is a phosphatidylcholine. 16. The pharmaceutical composition according to item 12, where the phospholipid is a sphingolipid. 17. The pharmaceutical composition according to item 12, where the neutral lipid is a triglyceride. 18. The pharmaceutical composition of claim 12, wherein the neutral lipid is cholesterol ester. 19. The pharmaceutical composition of claim 12, wherein the bile salt is selected from sodium cholate, sodium deoxycholate and sodium chenodeoxycholate. 20. The pharmaceutical composition according to claim 19, where the bile salt is sodium cholate. 21. The pharmaceutical composition according to item 12, containing phosphatidylcholine, triglyceride and sodium cholate. 22. The pharmaceutical composition according to item 21, containing phosphatidylcholine and triglyceride in a mass ratio of 93: 7. 23. The pharmaceutical composition according to claim 21, wherein sodium cholate, phosphatidylcholine and triglyceride have a total weight ratio of 13.1: 81.2: 5.7, respectively.

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