Anti-myalgic encephalomyelitis / chronic fatigue syndrome medicaments

Abstract

The invention relates to a combination of alpha-ketoglutaric acid or one of the salts or esters thereof and pyruvic acid or a pyruvate as a medicament for combating myalgic encephalomyelitis (ME) and / or chronic fatigue syndrome (CFS).

Description

MEDICINES FOR MYALGIC ENCEPHALOMYELITIS / CHRONIC FAILURE SYNDROME

[0001] The present invention relates to a drug for myalgic encephalomyelitis / chronic fatigue syndrome.

[0002] There is evidence that myalgic encephalomyelitis / chronic fatigue syndrome (ME / CFS) is caused by the activation of immune or inflammatory reactions via metal-induced lipid peroxidation of macrophages.

[0003] In Maes M, Kubera M, Uytterhoeven M, Vrydags N, Bosmans E. "Increased plasma peroxides as a marker of oxidative stress in myalgic encephalomyelitis / chronic fatigue syndrome (ME / CFS)", Med Sci Monit. 2011 Apr;17(4):SC11-5. doi: 10.12659 / msm.881699. PMID: 21455120; PMCID: PMC3539515, it was shown that the plasma peroxide concentration (TBARS; TBARS = thiobarbituric acid reactive substances) is significantly higher in ME / CFS patients than in healthy controls. There was also a trend toward significantly higher serum oxLDL antibodies in ME / CFS compared to controls. Both biomarkers thus contributed significantly to differentiating between ME / CFS patients and healthy controls.

[0004] There are substances that can prevent lipid peroxidation-mediated oxidation of LDL and TBARS, such as vitamin E and vitamin C.

[0005] It is not known that the combination of alpha-ketoglutarate (AKG) and pyruvate (Pyr) can also delay or inhibit metal-mediated lipid peroxidation.

[0006] Therefore, within the scope of the present invention, studies were conducted directly on isolated LDL, which was converted to oxidized LDL (oxLDL) and lipid radicals (TBARS) using copper ions. This was done in the absence and presence of the individual components AKG, Pyr, and the combination of both (AKG+Pyr). Surprisingly, it was found that the combination of AKG+Pyr significantly reduced lipid peroxidation. Subsequently, a clinical trial was conducted to determine whether this combination of alpha-ketoglutaric acid and pyruvate was effective against myalgic encephalomyelitis (ME) and / or chronic fatigue syndrome (CFS). The result of this study was that it was highly effective.

[0007] The present invention therefore relates to the combination of alpha-ketoglutaric acid or one of its salts or esters and pyruvic acid or a pyruvate as a medicament for the treatment of myalgic encephalomyelitis (ME) and / or chronic fatigue syndrome (CFS). Preferably, the molar ratio of alpha-ketoglutaric acid or one of its salts or esters to pyruvic acid or a pyruvate is between 1:1 and 10:1.

[0008] Any salts and esters are suitable: alkali, alkaline earth, transition metals up to creatine and arginine alpha-ketoglutarate.

[0009] The present invention is explained in more detail with reference to the experimental descriptions and the accompanying drawings. These show: the oxLDL values ​​before and after oxidation of LDL, wherein the oxidation was carried out without and with the addition of AKG, Pyr and AKG+Pyr; the TBARS values ​​after this experiment; and the effect of AKG+Pyr on fatigue.

[0010] Before starting the metal oxidation of LDL, it was dialyzed with 0.01 mol / l PBS, pH 7.4, and carefully degassed. After determining the protein content using the BCA method, an initial oxidation was carried out at 37 °C with 10 µmol / l CuSO₄. At intervals between 0 and 6 hours, the reaction was stopped by adding a stop solution to achieve a final concentration of 2.7 mmol / l EDTA.

[0011] The samples were saturated with nitrogen and stored at 4 °C.

[0012] To determine the degree of LDL modification, lipid peroxides were measured in all samples using a spectrophotometric test with CHOD iodide color reagent (Merck) at 365 nm, and carbonyl proteins were determined using an oxLDL assay.

[0013] It was shown that both the TBARS and the oxLDL of 4h Cu 2+ -oxidized LDL served as a good comparison for the following measurements:

[0014] LDL was again oxidized with 10 µmol / l CuSO4, without additives or in the presence of alpha-ketoglutarate, pyruvate, or with the combination of alpha-ketoglutarate:pyruvate (in a molar ratio of 3+1). Afterwards, both TBARS and oxLDL were measured again.

[0015] Figure 1 shows the oxLDL content measured at an absorption of 450 nm. It shows that oxLDL alone exhibits a threefold increase at 4 h compared to native LDL (Nat.LDL), which appears highly significant (p<0.001). In the presence of alpha-ketoglutarate (AKG) and also of pyruvate (Pyr), the oxidation of LDL was (only) more than twice as high compared to native LDL, with lower significance (p<0.01). The increase in oxLDL after 4 h in the presence of the mixture of AKG+Pyr showed the smallest increase compared to native LDL, with the lowest significance (p<0.05).

[0016] Comparing the results with 4h oxLDL (instead of Nat.LDL), the mixture of AKG+Pyr showed the best effect in reducing oxLDL production, with a high significance of p<0.01; the individual substances AKG and Pyr showed less significant effects (p<0.05). However, a significantly lower value was also found for the combination of AKG+Pyr compared to AKG or Pyr alone (p<0.05).

[0017] The determination of TBARS (cholesterol) showed the same trend, see [reference]. The mixture of AKG+Pyr showed the lowest value compared to 4h oxLDL (p<0.01), while AKG and Pyr were less low, but still significantly lower than 4h oxLDL (p<0.05).

[0018] Furthermore, a significant difference was found between AKG or Pyr alone and the mixture of AKG+Pyr with p<0.05.

[0019] It can therefore be shown that the combination of AKG+Pyr shows the best effect in preventing the oxidation of lipids, both oxLDL and TBARS.

[0020] There is evidence that myalgic encephalomyelitis / chronic fatigue syndrome (ME / CFS) is characterized by the activation of immune, inflammatory, oxidative and nitrosative stress (IO&NS).

[0021] The present study was conducted to investigate whether ME / CFS is associated with elevated levels of plasma peroxides and oxidized LDL (oxLDL), two biomarkers for oxidative stress.

[0022] Materials / Methods: Pooled plasma from ME / CFS patients and pooled blood plasma from healthy volunteers were purchased for analysis. The severity of ME / CFS was assessed using the Fibromyalgia and Chronic Fatigue Syndrome (FF) rating scale. Results: Plasma peroxide levels were significantly higher in ME / CFS patients (261 ± 35.2 µmol / L, p < 0.01) than in healthy controls (140 ± 11.0 µmol / L). A commercially available spectrophotometric test kit was used: Peroxides are determined via a reaction of peroxidase with peroxide followed by substrate conversion with TMB. After adding the stop solution, the chromogenic liquid is photometrically measured at a wavelength of 450 nm. Quantification is performed using a portable calibrator. There was also a significantly higher plasma OxLDL level in ME / CFS (777.1±62 ng / ml; p<0.01) than in control subjects (240.4±28.6 ng / ml).Measurement principle: The test is based on the classic sandwich ELISA technique. In the first incubation step, the target antigen is bound by immobilized polyclonal antibodies. All unbound substances are removed in a wash step. In the second incubation step, a peroxidase-labeled antibody is added. After another wash step to remove unbound components, the peroxidase substrate tetramethylbenzidine (TMB) is added. The enzyme reaction is stopped by the addition of acid. The resulting chromogenic compound is measured photometrically at 450 nm. The color intensity is directly proportional to the concentration of the measured analyte. The sample concentration can be determined using a standard curve – optical density (absorption at 450 nm) versus standard concentration. Both biomarkers significantly contributed to the differentiation between ME / CFS patients and healthy controls.Both plasma PerOx levels and plasma OxLDL levels were significantly associated with one of the FF symptoms. Conclusions: The results show that ME / CFS is characterized by increased oxidative stress.

[0023] A concentrate containing arginine AKG salt and creatine pyruvate salt (1.8 g arginine AKG and 0.5 g creatine pyruvate daily) was administered to subjects for three weeks. Before and after administration, a fatigue score questionnaire (according to Stein KD, Jacobsen PB, Blanchard CM, Thors C. "Further validation of the multidimensional fatigue symptom inventory-short form" J Pain Symptom Manage. 2004 Jan;27(1):14-23) was completed. After a washout period, the subjects were surveyed again, with no changes to their lifestyle. The results are shown.

[0024] The Kruskal-Wallis test is a non-parametric test for comparing three or more groups with a metric or ordinal dependent variable. It is frequently used when the assumptions for an analysis of variance (e.g., normality, equal variances) are not met.

[0025] The precise nature of these differences (i.e., which groups differ from which others) requires additional post-hoc Bonferroni tests. Test statistic Standard error Std. Test statistic p Adjusted p Total fatigue before - Total fatigue after 22.18 5.18 4.28 <0.001 <0.001 Total fatigue before - Fatigue after washout 8.23 ​​5.75 1.43 0.15 20.45 7 Total fatigue after - Fatigue control -13.95 5.75 -2.43 0.01 50.04 6 Adjusted p: Values ​​adjusted with Bonferroni correction.

[0026] The results show that daily oral administration of the combination of alpha-ketoglutarate and pyruvate significantly improved fatigue after three weeks compared to baseline. After a washout period, the values ​​worsened again; while they were better than before treatment, the improvement was not significant. Interpretation:

[0027] The results of the Kruskal-Wallis test, with a chi-squared value of 18.67, a df of 2, and a p-value of <0.001, indicate a statistically significant difference between the three compared groups. Therefore, the null hypothesis is rejected based on the available data. Post-hoc test

[0028] Using a Dunn-Bonferroni test, the groups were compared pairwise to find out which ones differed significantly.

[0029] The Dunn-Bonferroni test revealed that the pairwise group comparisons of total fatigue before - total fatigue after treatment had an adjusted p-value less than 0.001 and total fatigue after treatment - fatigue after washout had an adjusted p-value just less than 0.05, and thus, based on the available data, it must be assumed that these groups differ significantly pairwise.

[0030] Maes M, Kubera M, Uytterhoeven M, Vrydags N, Bosmans E. "Increased plasma peroxides as a marker of oxidative stress in myalgic encephalomyelitis / chronic fatigue syndrome (ME / CFS)", Med Sci Monit. 2011 Apr;17(4):SC11-5. doi: 10.12659 / msm.881699. PMID: 21455120; PMCID: PMC3539515 nplcit2: Stein KD, Jacobsen PB, Blanchard CM, Thors C. "Further validation of the multidimensional fatigue symptom inventory-short form" J Pain Symptom Manage. 2004 Jan;27(1):14-23

Claims

Combination of alpha-ketoglutaric acid or one of its salts or esters and pyruvic acid or a pyruvate as a medicinal product for the treatment of myalgic encephalomyelitis (ME) and / or chronic fatigue syndrome (CFS). Combination according to claim 1, characterized in that the molar ratio of alpha-ketoglutaric acid or one of its salts or esters to pyruvic acid or a pyruvate is between 1:1 and 10:1.

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