KR100372672B1 - Compositions and methods useful for the prevention and treatment of endotoxin-related diseases - Google Patents

Abstract

Methods for treating and preventing endotoxin-induced addiction are disclosed. This is done by administering to the patient a composition containing phospholipids. The composition does not contain proteins and peptides and may contain triglycerides, other polar or neutral lipids, bile acids or bile salts.

Description

Compositions and Methods Useful for the Prevention and Treatment of Endotoxin Related Diseases

Endotoxin shock is most gram-negative bacteria (eg, Escherichia coli); Salmonella tymphimurium] is often a fatal disease caused by lipopolysaccharide (LPS) released from the outer membrane. The structure of the bacterial LPS is very well known, and a unique molecule called lipid A is bound to the acyl chain through its glucoseamine backbone. See Raetz, Ann. Rev. Biochem. 59: 129-170, 1990 in this regard.

Lipid A molecules function as membrane anchors of lipopolysaccharide (“LPS”) and are LPS implicated in the expression of endotoxin shock. It should be pointed out that LPS molecules are characterized by lipid type A structures and polysaccharide moieties. This latter part should retain the general structural basis properties of endotoxin, although the details of the molecule may differ in different LPS molecules. It would not be accurate to claim that LPS molecules are identical among bacteria (see Raetz et al., Supra). It is common in the art to refer to various LSP molecules as "endotoxins", which term is later used herein to generically refer to LPS molecules.

The disclosure of US Pat. No. 5,128,318 is incorporated herein by reference, in which a reconstituted particle containing both apo lipoproteins associated with HDL and lipids capable of binding to and inactivating endotoxins mitigates toxicity due to endotoxins. It is indicated that it can be used as an effective substance to make it.

US Pat. No. 5,344,822, International Patent Application No. PCT / US93 / 07453, and US Patent Application No. 08 / 288,568, incorporated herein by reference in their entirety as applications related to this application, are toxic due to endotoxins using various other materials. It is disclosed that can be treated. Specifically, it has been found that apo lipoproteins are not required for reconstituted particles, and that the reconstituted particles may contain peptides and lipids that are not apo lipoproteins.

The inventors have also found that toxicity due to endotoxin can be treated by sequential administration of lipids following peptides or apolipoproteins. After sequential administration, these components associate with the reconstituted particles and then remove the endotoxin.

It has also been found that at least some individuals inherently possess apolipoproteins above normal levels, thereby enabling effective endotoxemia treatments by administering reconstituted particles that do not contain apolipoproteins or peptides but contain the disclosed lipids.

In addition, the inventions disclosed herein include the use of the reconstituted particles and components discussed herein for preventing toxicity due to endotoxin by administering a prophylactically effective amount to a subject in need thereof. The subject includes a patient recovering from a surgical operation or suffering from an infection. These patients have very low levels of plasma HDL, often only 20% of normal levels. In such cases, early prevention using HDL to compensate for this degradation is highly desirable.

Very surprisingly, it has also been found that phospholipids can be used alone or in combination with additional substances such as neutral lipids, cholates, etc. as an active agent to alleviate and / or prevent endotoxemia. It is particularly preferred to use phosphatidylcholine (hereinafter referred to as "PC") alone or in combination with other phospholipids, such as spinolipids, in compositions substantially free of peptides and proteins such as apolipoproteins or peptides derived therefrom. When the composition is injected intravenously into the bolus, neutral lipids such as mono-, di- and triglycerides can be used with phospholipids as long as the total amount of neutral lipids is below a certain weight percentage. When used in other dosage forms such as continuous intravenous infusion, the weight percentage is not critical but is required.

Particularly preferred embodiments of the present invention include emulsions in which bile acids or bile salts are used with phospholipids and neutral lipids.

The present invention demonstrates the efficacy of treating endotoxins of bile acids and bile salts which are cholates. These bile acids may be used alone or in combination with one or more phospholipids (eg phosphatidylcholine) and / or neutral lipids (eg triglycerides).

The invention is described in more detail below.

The present invention relates to the treatment of endotoxin-related endotoxins. More particularly, the present invention relates to the treatment of such addiction through the administration of various compositions that act to neutralize and / or eliminate endotoxins from an organism and to prevent the use of these compositions.

1A and 1B show the results obtained by testing various compositions in a model measuring neutralization of endotoxin through the measurement of TNF release in a whole blood model of humans. Figure 1A shows the role of protein, Figure 1B shows the role of phospholipids. The compositions tested included neutral lipoproteins (VLDL, LDL, HDL), reconstituted HDL ("R-HDL"), and intra lipid (INTRALIPID®) compositions, and emulsions containing phospholipids and proteins.

2A and 2B compare the roles of triglycerides (neutral lipids) and phosphatidylcholine (phospholipids) in the same model.

3 is this. The 55% lethal model administered with Coli LPS is used to show information about the toxicity associated with the administration of the various PC and PC / TG compositions in the mouse model.

FIG. 4 shows data almost equivalent to that obtained in the whole blood assay of humans, although unesterified cholesterol, spinomyelin, or a mixture of both was used with phospholipids instead of triglycerides.

5A and 5B show comparable results to FIGS. 1A and 1B, except for mixing phospholipids, unesterified cholesters, and / or spingomyelin with neutral lipids triglycerides or esterified cholesterols. Indicates.

FIG. 6 compares the results obtained from emulsions containing cholester esters and triglycerides in a mouse model in vivo.

FIG. 7 shows the theoretical amounts and toxic thresholds of triglycerides released into the blood upon administration of various TG containing compositions. "TPN" refers to "total parenteral nutrition" and "RI" refers to a composition according to the invention.

Detailed Description of Embodiments of the Invention

<Example 2>

Factors affecting LPS-mediated stimulation of TNF-α while preserving the interaction between plasma proteins and cellular components of the blood can be appropriately studied in human, whole blood system in vitro. This system was used to identify which lipoprotein components are important for neutralizing LPS.

Substances tested were high density lipoprotein (R-HDL), natural plasma lipoprotein (VLDL, LDS, HDL), lipoprotein deficient serum (LPDS), and triglyceride rich emulsion 20% intralipid (a mixture of triglycerides and phospholipids).

Blood was collected in Hank's Balanced Salt Solution (hereinafter referred to as "HBSS") or in tubes coagulated with heparin and diluted with the material to be tested dissolved in HBSS. The material obtained was transferred to a Starstedt tube (250 ul / tube). LPS was dissolved in saline containing 10 mM HEPES and pyrogen free, and added to a final concentration of 10 ng / ml (2.5 ul). After 4 hours of incubation at 37 ° C., the tubes were cooled to 4 ° C. and then centrifuged at 10,000 × g for 5 minutes. Supernatants were collected and analyzed using a commercial ELISA to determine TNF-α.

Table 1 below compares the composition of the materials tested. 1A and 1B show the results. Data was plotted by plotting the amount of TNF-α produced against the concentrations of added protein (FIG. 1A) and phospholipids (FIG. 1B). In order to show all the wide concentrations used, 10 ° is shown in logarithm units at 1 mg / ml. All whole blood cultures were 10 ng / ml of E. coli. Coli 0111: B4 LPS was contained and supplemented with one of the compositions as indicated by symbols in FIGS. 1A and 1B.

The fact that the substances are very similar in terms of efficacy when plotting phospholipid content (FIG. 1B) but different in terms of efficacy when plotting protein content (FIG. 1A) suggests that phospholipid is an important component. This is confirmed by the finding that protein-free lipid emulsions are more effective than native HDL, but less effective than R-HDL. Protein does not appear to be important for neutralization.

Composition of Natural Lipoproteins and Reconstituted HDL

<Example 3>

As a next step, protein-free lipid emulsions containing different amounts of neutral lipids were tested in whole blood of humans. The same in vitro human whole blood assay as shown in Example 2 was used.

All particles described herein are prepared through the same manner, including mixing, weighing and placing into the flask phospholipids, spinomyelin or phosphatidylcholine, triolein and / or non-esterified cholesterol esters dissolved in chloroform. It was. Vitamin E (0.02% w / v) was added as antioxidant. Subsequently, nitrogen or argon gas was blown onto the sample to prepare a dry lipid film. A large amount of nonpyrogenic saline was then added to the flask and mixed in a vortex mixer until all lipids were suspended. The solution was then homogenized in a high pressure homogenizer. Samples containing phosphatidylcholine (PC) with or without triolein were circulated 10 times at 20,000 psi on a homogenizer. Samples containing cholesterol esters with one or more other lipids were cycled 15-20 times at 30,000 psi. The homogenized solution was filtered through a 0.45 μm syringe filter and the filtrate was stored at room temperature until use (within 3 days). These results are shown in Figures 2A and 2B. In this study, LPS-dependent TNF-α production was plotted against the concentration of added triglycerides (FIG. 2A) or phospholipids (FIG. 2B). As indicated, the composition contained 7% by weight triglycerides (“TG”), 45% TG, 89% TG, 94% TG, R-HDL, or phospholipids without TG (shown only in FIG. 2B). The 89% TG composition is a 10% intralipid formulation and 94% TG represents 20% intralipid. Egg phosphatidylcholine (PC) and triolein were used in all other tests.

These results show that the protein-free compositions are very different when compared via triglyceride content. They are very similar when tested via phospholipid (PC) content. This is particularly valid for phospholipids alone but less effective than emulsions containing less than 45% TG, confirming the role of phospholipids.

<Example 4>

Subsequently, in vivo experiments were conducted in mouse models that are believed to be a reliable way to predict efficacy in humans.

In these experiments, the formulations described in Example 3 and other (pure phosphatidylcholine, 7% TG, 25% TG, 45% TG, 71%) were sufficient to provide a phospholipid dose (200 mg / kg or 400 mg / kg). TG, 81% TG, 89% TG, 94% TG) or saline control were injected into mice in bolus form with 25 mg / kg of E. coli 0111: B4 LPS. The control group was injected intravenously with a volume of physiological saline sufficient to correspond to the volume of the emulsion. Survival after 72 hours is shown in FIG. 3. Only 155 of 344 control groups survived.

PC alone had a moderate protective effect that was not statistically significant at the 95% confidence level, but the 7%, 45% and 71% TG compositions significantly improved survival. The 80% and 89% TG compositions had some effect, but 94% TG reduced survival.

When the dose was increased to provide 400 mg / kg of PC, both 89% and 94% TG emulsions significantly reduced survival time, possibly due to TG toxicity, as described below.

Example 5

The content described in Examples 2-4 demonstrated that phospholipids are useful active agents in inhibiting endotoxemia. The fact that nonpolar lipids other than triglycerides can form phospholipids and emulsions other than PCs suggests that something else could be tried. Examples are spingomyelin (another phospholipid) and nonesterified cholesterol (polar neutral lipids) and mixtures thereof. Esterified cholesterol (non-polar esters), squalene (hydrocarbons) and vitamin E (non-polar antioxidants) can also be used. A series of experiments were planned to test them using the human whole blood assay of Example 2 and the mouse survival assay of Example 4.

Emulsions were prepared using the method described above using phosphatidylcholine containing 10% by weight of pure phosphatidylcholine, 10% by weight of non-esterified cholesterol, 10% by weight of spinomyelin, or a mixture of both. The emulsion was added to whole blood at a concentration of 100 mg / dl PC and 10 ng / ml LPS. The mixture was incubated and TNF-α release was measured.

The results are shown in FIG. TNF-α production was substantially reduced when only PC was used. Emulsions containing non-esterified cholesterol, spinomyelin, or a mixture of both were also inhibitory to TNF-α release.

<Example 6>

Whole blood analysis was used to determine the effect of unesterified cholesterol and / or spingomylin on emulsions containing neutral lipids. At this time, the emulsion was added at 100 mg / dl PC. Various compositions (weight / weight) are shown in the table below.

5A and 5B show the results. PC emulsions prepared using neutral lipids with or without additional polar lipids showed inhibition. LPS concentration was used 10 ng / ml, a clinically problematic endotoxin concentration. Cholesterol ester containing emulsions are less effective than TG containing emulsions, but emulsions containing non-esterified cholesterol as well as emulsions containing no esters did not inhibit TNF-α. Spinomyelin was added to the emulsion to improve the inhibition of TNF-α production.

<Example 7>

Cholesterol ester containing emulsions were tested in a biometric model (ie, those used in Example 4) using lethal amounts of endotoxin. Emulsions are prepared using PC and TG, or PC and cholesterol esters (CE), 25 mg / kg Yi. A single bolus dose of 200 mg / kg PC was administered via the tail vein along with Coli 0111: B4LPS. In the control group, saline was injected intravenously in accordance with the volume of the emulsion.

The data in FIG. 6 compares the results from the CE and TG containing emulsions. Each emulsion was tested in at least 2 experiments with a total of 16 animals or more.

As shown, emulsions containing 7% or 45% CE (% by weight) significantly improved survival. Taken together with the results of Example 6, it can be seen that CE can produce an emulsion that replaces TG to neutralize endotoxin.

<Example 8>

Emulsions of phospholipids containing no protein and containing triglycerides effectively blocked TNF-α production promoted by LPS in whole blood. Theoretically, these emulsions would be effective in vivo if they could be safely administered at a dose that provides a protective effective concentration of phospholipids in plasma. The experiments using our R-HDL suggest that the minimum dose of phospholipids is approximately 200 mg / kg. Using this dosage and a plasma volume of 4.5% of body weight, the concentration of triglycerides expected in the plasma can be calculated after administration of a series of emulsions with increasing triglyceride content. This result is shown in FIG. 7 as a continuous line bending upward as the TG weight% increases. Plasma TG levels rarely exceed 1000 mg / dl in healthy adults even after fatty meals. Pancreatitis has been reported in patients with plasma TG above 2000 mg / dl [Farmer et al., Amer. J. Med. 54: 161-164 (1973); Krauss et al., Amer. J. Med. 62: 144-149 (1977); Glueck et al., J. Lab. Clin. Med. 123: 59-61. Plasma TG above 4000 mg / dl is extremely rare and causes serious concern. The last two limits are indicated by horizontal lines in the above figures. Administration of 10% or 20% intralipid at doses providing 200 mg / kg phospholipids is expected to raise plasma TG concentrations above the safety limits described above (see two reductions). In contrast, administration of emulsions containing 7%, 45%, 71% or 78% (left to right color squares) raised plasma TG to 136, 477, 1300 or 2000 mg / dl, respectively. Emulsions with a TG content of up to 50% are expected to be free of toxicity due to TG.

Example 9

The efficacy of the combination of phospholipids and bile acids (ie sodium cholate) was tested in the same type as the experiments performed in the above examples.

However, the method of preparing the formulation administered to the test animal was different.

In this example and the following examples, formulations were prepared using a Microfluidizer high pressure homogenizer. This device is easy to scale up.

Either liquid triolein or liquid soy triglycerides were weighed into an appropriate amount of water or water to which 9 mM, 18 mM or 36 mM sodium cholate was added. Solid granular phosphatidylcholine was weighed on a weighing paper and then slowly added to the solution with stirring. It took about 3 to 5 minutes to disperse the lipids. After dispersion, the material was poured into a microfluidizer. The device uses hydraulic pressure to operate the pump, which directs the two sample jets to each other. The pressure can be as high as 1.76 × 10 ⁶ g / cm ² (25,000 psi). The sample was homogenized by pressurizing the injection liquid through the + type orifice at the time of impact.

The sample is recycled through the microfluidizer, where a "one pass" is defined as the time taken to pump all the samples through the device. Samples were cycled 20 times to produce homogenized samples. Dextrose was added to a final concentration of 5%.

this. Endotoxin (40 mg / kg) purified from Coli 0111: B4 and an emulsion as discussed below (200 mg phosphatidylcholine / kg) are mixed at room temperature and immediately tail vein into C57BL6 / J mice (weight 19 to 30 g) Via intravenous injection. Mice injected with xyl salt alone received the same volume of sodium cholate as the cholate / EML (emulsion) formulation at the same choline salt concentration. Mice in the control group were injected with the same volume of 5% dextrose to meet plasma osmotic pressure.

The results are shown in the table below. The emulsion was a phosphatidylcholine / 7% triglyceride emulsion as described in the examples above. Sodium cholate, if used, was added at the concentrations indicated in the raw material prior to emulsifying the raw material.

TABLE 1

Mouse protection against lethal doses of lethal doses

For convenience, the weight percentage of the emulsion is as follows. When using 9 mM cholate, the weight percentages for the emulsions were 7% cholate, 6.1% triglycerides, and 86.9% phosphatidylcholine. At 18 mM cholate, the weight percentages were 13.1% cholate, 5.7% triglycerides and 81.2% phosphatidylcholine. For 36 mM cholate, the relative values were 23.2% cholate, 5% triglycerides and 71.8% phosphatidylcholine.

It should be noted that the amount of LPS administered in these experiments (40 mg / kg) was much higher than the amount used in the lethal experiments conducted previously. The intention of these high doses is to overwhelm all the protective effects attributable to phosphatidyl choline and / or triglycerine. Thus, the conclusion reached with these experiments is that there is a protective effect due to bile salts, sodium cholate.

Studies conducted with other bile salts and taurine containing bile salts are not described herein. Further examples of bile acids include allodeoxycholic acid, lithocholic acid, hydeoxyolic acid, hycolic acid, α, β and ω-muricolic acid, mudeoxycholic acid, ursodeoxycholic acid, ursokolic acid, and these All salts of, for example, its sodium salt or taurine or glycine conjugates. See, Hoffman et al., Supra.

<Example 10>

Further studies were performed using mice as subject animals, the first being survival studies.

In survival studies, subject animals were divided into four groups. In the first group, 5% dextrose solution was administered and used as a control. The second group was administered an emulsion of 93% (by weight) phosphatidylcholine and 7% (by weight) triglycerides prepared as described above. The emulsion contained 5% dextrose and soybean phospholipids at approximately 50 mg / ml lipid.

In groups 3 and 4, animals were injected with emulsions similar to those provided in group 2 and supplemented with 18 mM sodium cholate or 18 mM sodium deoxycholate. The protocol used in this experiment was the same as shown in Example 9.

Survival was measured 72 hours after dosing and summarized in the table below.

TABLE 1

Effect of Bile Acids in 7% Triglyceride Emulsion on 72-Hour Survival in Mice

Statistical significance of comparison between group survival was tested by the generalized Walcox method using a computer program. A comparison to Control 1 is described in column "1" and a comparison to Group 2 animals treated with 7% emulsion is described in column "2" and a comparison to Group 3 animals treated with emulsion plus sodium cholate Described in the "3" column.

Survival and statistical analysis both show clear and unexpected superiority of formulations containing bile salts.

<Example 11>

Another series of experiments used a rabbit model. In this model, the release of TNF (Tumor Necrosis Factor) -α was measured.

The rabbits were divided into 3 groups and administered a 5% dextrose solution, an emulsion of phospholipids and triglycerides (93% / 7%) discussed above or a 93% / 7% emulsion containing 18 mM cholic acid. All emulsions were adjusted to 5% dextrose content as in Example 10. Two hours after the priming bolus of the emulsion was administered to the rabbit, 100 μg of E. coli. Coli 0111: B4 LPS was stimulated. Following administration of the priming bolus, the formulations were administered intravenously to allow continuous maintenance infusion of 50 mg of lipid per kg of body weight per hour. Intravenous administration was continued for 3 hours after stimulation.

Blood from rabbits was taken every hour over baseline, 30 minutes after priming bolus administration and 5 hours post administration.

In the table below, the peak values of TNF-α are shown. They appeared 2 hours after endotoxin administration.

Statistical significance was measured using a known Student's test. As shown in the table, TNF-α levels decreased significantly after administration of 18 mM cholic acid.

TABLE 2

Effect of Emulsion on TNF-α Production in Rabbits

The above embodiments detail the present invention comprising alleviating or preventing endotoxin in a patient by administering an effective amount of phospholipid to which endotoxin binds. The combination of phospholipids and endotoxins is removed from the patient through standard biological methods known to those skilled in the art who are familiar with how to remove lipoprotein particles. When endotoxins bind to phospholipids, endotoxins are inactivated.

The examples show that the administration of one of the cholanic acid or choline salt series such as bile acid or bile salt can be used for the same purpose as phospholipids, i.e. alleviation or prevention of endotoxemia, thus, bile acid / bile salts and phospholipids Compositions containing either or both of them and without peptides and proteins can be used for the treatment of endotoxemia. Cholanic acid is disclosed, for example, in Hofmann, Hepatology 4 (5): 4S-14S (1984), which is incorporated herein by reference. Of particular note are the FIGS. 1 and 2, 5S planes, which are incorporated herein by reference, which show the structural features of cholanic acid.

The patient to be treated is preferably a human, but the practice of the present invention is equally applicable to the veterinary scope.

"Relax" means a treatment that alleviates the burden of endotoxin caused by any of a variety of endotoxins produced by, for example, Gram-negative bacteria (S. tymphimurium, E. coli, etc.). Prophylaxis may be achieved by administering the agent at or at the point where the subject may be exposed to endotoxin. Typically this occurs during a surgical operation. Thus, the active ingredient may be administered prior to this procedure to a patient who will be undergoing a surgical procedure.

The effective amount of the phospholipid and bile acid combination required for the treatment of the subject may vary. In the context of severity or prophylaxis of endotoxins, the amount may be reduced or increased depending on the risk, but generally a dose of about 200 mg up to about 800 mg total phospholipids per kg body weight of the subject is preferred. In the case of cholanic acid and salts such as bile acids and salts thereof, dosages of from about 10 mg to about 300 mg, more preferably from 15 mg to about 275 mg per kg body weight are used.

It is preferred to administer the bile acids / bile salts and phospholipids in compositions that also contain neutral lipids, but this is not necessary as emulsions of phospholipids without neutral lipids can also be intended. Preferred administration of a combination of phospholipids is due to the fact that the neutral lipids and phospholipids associate to form particles that differ from lipoproteins in that there is no protein or peptide component that is naturally always present in the lipoprotein.

A particularly preferred form of treatment is that the phospholipid is phosphatidylcholine such as egg yolk phosphatidylcholine, phosphatidylcholine based on soybean or spinolipid. In the case of bile acid / bile acid salts, preference is given to cholic acid and / or salts thereof such as sodium cholate, sodium deoxycholate and sodium kenodeoxycholate. As for neutral lipids, it is preferred to use cholester esters or triglycerides, but other neutral lipids such as squalene or other hydrocarbon oils, di- and mono-glycerides and antioxidants such as vitamin E can also be used.

The form in which the composition may be administered may vary, but bolus or other intravenous form is particularly preferred. If the bolus form is used and the composition contains triglycerides, some care must be taken when administering. It is known that triglycerides are toxic when administered in excessive amounts. However, those skilled in the art can readily prepare the composition such that the risk of triglyceride toxicity is reduced or eliminated. In general, when using the bolus form, the composition should contain up to about 80%, preferably up to 70% by weight of triglycerides or other neutral lipids. Most preferably, the composition should contain up to about 50% by weight of neutral lipids when administered in bolus.

However, the use of non-bolus forms, such as intravenous dosage forms other than bolus, reduces the risk of poisoning. Nevertheless, it should be noted that the above-mentioned ranges are preferred for, but are not limited to, intravenous or other dosage forms. For bile acids and bile salts, the dosage is preferably from about 25 mg to about 500 mg per kg body weight, particularly preferably from about 50 mg to about 100 mg per kg body weight. For phospholipids, dosages of from about 100 mg to about 1000 mg per kg of body weight are preferred. However, dosages generally vary with the subject and form of administration.

As described above, preparations without proteins and peptides require the presence of at least one phospholipid or bile acid / bile acid salt. For phospholipids, it is preferred to have at least one neutral lipid such as triglycerides, diglycerides, or monoglycerides. . The composition may be selected from sterols (eg, cholesterol, β-sitosterol), esterified or non-esterified lipids (eg, cholesterol esters or non-esterified cholesterol), hydrocarbon oils such as squalene, antioxidants such as vitamin E, and the like. May contain additional substances, but these are not essential. It is also possible to use two or more phospholipids and / or two or more neutral lipids in all of the above formulations. When using a combination of neutral lipids and phospholipids, the neutral lipids should be present from about 3% to about 50% by weight relative to the total amount of lipids in the composition.

In the case of bile acids / bile salts, they can be used separately or in combination with phospholipids, neutral lipids or both. With respect to these additional substances (eg phospholipids and neutral lipids), the preferred species are as discussed and described above. Optional additional ingredients include those described above.

Some of the present invention are compositions useful for treating endotoxins. One aspect of this aspect of the invention is a composition containing at least one bile acid / bile acid salt, phospholipid and neutral lipid, respectively, wherein the composition as a whole contains an active component in an endotoxin alleviating amount. The composition preferably contains about 5% to about 30% by weight of bile acid / bile acid salts, about 3% to about 50% by weight of neutral lipids, and about 10% to about 95% by weight of phospholipids. Particularly preferably the composition is a composition containing from about 10 to 15% by weight bile acid / bile acid salt, from about 5% to about 10% by weight neutral lipids, with the remainder of the composition being phospholipids.

It should be appreciated that the weight percentages are for compositions of three components. If the three component system is combined with, for example, a carrier, an adjuvant, any of the components discussed above, the weight percentages for the total composition will be lowered. Note that such therapeutic compositions do not contain proteins and peptides.

For compositions that do not contain bile acids or bile salts, the composition without proteins and peptides preferably contains about 3% to about 50% by weight neutral lipids, with the remainder being one or more phospholipids. Neutral lipids are preferred triglycerides, but may be any of the additional neutral lipids discussed above. Further, phospholipids are preferably phosphatidylcholine.

Other aspects of the invention will be apparent to those skilled in the art and need not be repeated herein.

It is to be noted that the foregoing description and examples are exemplary, and are not intended to limit the invention, and that other aspects within the spirit and scope of the invention may be proposed by one of ordinary skill in the art.

Claims (19)

A pharmaceutical composition for the treatment or prophylaxis of a person or animal suffering from endotoxin, comprising cholanic acid or choline salts, phospholipids and triglycerides. The pharmaceutical composition of claim 1 comprising 3 to 80% by weight of neutral lipids. A pharmaceutical composition according to claim 1 or 2 comprising 3 to 70% by weight of neutral lipids. A pharmaceutical composition according to claim 1 or 2 comprising 3 to 50% by weight of neutral lipids. The pharmaceutical composition according to claim 1 or 3, wherein 3 to 50% of the composition lipid content is neutral lipid. Persons or animals with endotoxins, including cholanic acid or cholate, phospholipids and triglycerides, with neutral lipids present in an amount of from 3 to 50% by weight of the total amount of fat and suitable for intravenous administration and lacking proteins and peptides Pharmaceutical compositions for the treatment or prophylaxis. The pharmaceutical composition according to claim 6, wherein the cholanic acid or choline acid salt is a bile acid or bile acid salt. For the treatment or prophylaxis of a person or animal suffering from endotoxemia, comprising 5 to 30% by weight of bile acids or bile salts, 3 to 50% by weight of neutral lipids, and 10 to 95% by weight of phospholipids and free of proteins and peptides Pharmaceutical composition. The pharmaceutical composition according to claim 8, wherein 10 to 15% by weight is bile acid or bile salt, 5 to 10% by weight is neutral lipid and the remainder is phospholipid. The pharmaceutical composition according to claim 8 or 9, which is suitable for intravenous administration. The pharmaceutical composition according to any one of claims 6 to 9, wherein the phospholipid is phosphatidylcholine. The pharmaceutical composition according to any one of claims 6 to 9, wherein the phospholipid is spinolipid. The pharmaceutical composition according to any one of claims 6 to 9, wherein the neutral lipid is triglyceride. The pharmaceutical composition of claim 6, wherein the neutral lipid is a cholesterol ester. The pharmaceutical composition of claim 7, wherein the bile acid salt is selected from sodium cholate, sodium deoxycholate and sodium kenodeoxycholate. The pharmaceutical composition of claim 15, wherein the bile salt is sodium cholate. 17. The pharmaceutical composition of any one of claims 6-9 and 16, comprising phosphatidylcholine, triglycerides and sodium cholate. 18. The pharmaceutical composition of claim 17, wherein the phosphatidylcholine: triglyceride is present in a weight ratio of 93: 7. 18. The pharmaceutical composition of claim 17, wherein the total weight ratio of sodium cholate, phosphatidylcholine and triglyceride is 13.1: 81.2: 5.7.

Images