J Diabetes Res. 2013;2013:763125.
The novel oral drug Subetta exerts an antidiabetic effect in the diabetic Goto-Kakizaki rat: comparison with rosiglitazone.
Bailbé D, Philippe E, Gorbunov E, Tarasov S, Epstein O, Portha B.
Laboratoire B2PE (Biologie et Pathologie du Pancréas Endocrine), Unité BFA (Biologie Fonctionnelle et Adaptive), Equipe 1, Université Paris-Diderot et CNRS EAC 4413, 4 rue Lagroua Weill Hallé, Paris Cedex 13, France.
The aim of the present study was to evaluate the potential antidiabetic effects of two-component drug Subetta and its components (release-active dilutions of antibodies to β-subunit insulin receptor (RAD of Abs to β-InsR) and to endothelial nitric oxide synthase (RAD of Abs to eNOS)) in Goto-Kakizaki (Paris colony) (GK/Par) diabetic rats. Subetta was administered orally for 28 days once daily (5 mL/kg) and compared to its two components (2.5 mL/kg), Rosiglitazone (5 mg/kg), and vehicle (5 mL water/kg). At day 28, fasting plasma glucose levels were significantly decreased only in Subetta and Rosiglitazone groups as compared to vehicle (P < 0.01): 147 ± 4 mg/dL and 145 ± 4 mg/dL and 165 ± 4 mg/dL, respectively. The data of glucose tolerance test showed that Subetta and RAD of Abs to β-InsR (similar to Rosiglitazone) prevented significantly (P < 0.01) the age-related spontaneous deterioration of glucose tolerance as seen in the control group. Subetta and RAD of Abs to β-InsR did not significantly modify the glucose-induced insulin secretion. Chronic administration of Subetta and RAD of Abs to β-InsR improves glucose control, to an extent similar to that of Rosiglitazone. We hypothesize that Subetta and RAD of Abs to β-InsR mostly act via an insulin-sensitizing effect upon target tissues.
Glucose control and OGTT:
Chronic treatment with Subetta and RAD of Abs to β-InsR, but not with RAD of Abs to eNOS, prevented diabetes progression and significantly decreased plasma glucose as compared with baseline values (153 ± 4 dg/mL versus 168 ± 8 dg/mL (p < 0.01) and 147 ± 4 dg/mL versus 167 ± 3 dg/mL (p < 0.001), respectively) and with H2O control group as well in case of Subetta (147 ± 4 dg/mL versus 165 ± 4 dg/mL (p < 0.01)).
Oral glucose tolerance tests showed that glucose intolerance spontaneously deteriorated with aging (at least within the time-window 10–14 wks.) in the male GK/Par rats in both control groups (H2O and CMC). Animals in RAD of Abs to β-InsR, Subetta, and Rosiglitazone groups exhibited significantly lower postoral glucose loading glucose levels than those in the controls: AUC glucose variations during the 28-days period were lower by 41% (p < 0.001), 59% (p < 0.05), and 41% (p < 0.05) as compared to respective controls (H2O and CMC) (Figure 1). This establishes that both RAD of Abs to β-InsR and Subetta exert positive long-term effect upon glucose homeostasis in GK/Par rat model of type 2 of diabetes, which is comparable with Rosiglitazone effect. Herewith Subetta effect exceeds RAD of Abs to β-InsR effect.
Efficacy in experimental model of streptozotocin-induced diabetes:
Previously, in experimental model of streptozotocin-induced diabetes Subetta showed pronounced antihyperglycemic activity, which is comparable to that of the reference drug Rosiglitazone: Subetta decreases high plasma levels of glucose, urine levels of ketone bodies and improves glucose uptake in peripheral tissues .
We were the first to discover physical phenomenon of release-activity. Technological treatment of the starting substance as consecutive multiple decrease in its concentration results in production of fundamentally new activity absent in the starting substance which we named release-activity. Release-activity is exhibited as the ability of technological treated products to modify physical, chemical and biological properties of the starting substance [2, 3].
Mechanism of action:
The findings of the current study suggest that Subetta action is mostly at the level of insulin action on the target tissues. Taking into account that Subetta belong to the class of novel drugs and shares its common properties [2, 3], such mechanism might be carried out by modulating effect of Subetta on the β-subunit of the insulin receptor regulating the insulin receptor’s kinase activity and consequently activating receptor-associated signaling pathways. Partly direct action of Subetta on insulin receptor has been recently confirmed in vitro, where it was shown that Subetta significantly stimulates adiponectin production by mature human adipocytes in the absence of insulin (Figure 2) , which is known to enhance adiponectin regulation and secretion selectively in adipocytes .
Figure 1. Time-related variations of AUCglucose values in each group between Day 0 (d0) and Day 28 (d28). Data are expressed as means±S.E.M. from 12 observations/group. Mann-Whitney test for unpaired data was used for statistical analysis. ** – p <0.01 vs d0-H2O-treated GK/Par group; * – p <0.05 vs d0-CMC-treated GK/Par group. CMC – carboxy-methyl-cellulose; Rosi – Rosiglitazone; AUC – incremental integrated blood glucose values (AUC glucose; g/L/120min) (sum of value at tn− value to, for tn = 5, 10, 15, 30, 60, and 120 min).
Figure 2. Effect of Subetta and rosiglitazone on human adipocyte adiponectin secretion. Incubation of mature human adipocytes with Subetta for 72 hours resulted in a statistically significant increase in adiponectin concentration in the culture medium. Nonspecific controls, RAD of Abs to R-CBI and RAD of RbS, did not significantly affect adiponectin secretion, resulting in adiponectin levels comparable to background values of the negative controls, placebo, purified water, and 0.1% dimethyl sulfoxide (DMSO). The reference drug Rosiglitazone stimulated adiponectin secretion but to a lower level compared to that of Subetta and its effect was not significant in comparison with DMSO value. Data are expressed as means±SD from 4-6 replicas. One-way analysis of variance (ANOVA) followed by Tukey HSD test was used for statistical analysis. *** – p<0.001 vs either RAD of Abs to R-CBI, or RAD of Rbs, or Placebo, or Purified water, or DMSO (0.10%); # – p<0.05 versus Rosiglitazone (1 uM). RAD of Abs to R-CBI – release-active dilutions of antibodies to cannabinoid receptor type I; RAD of RbS – release-active dilutions of rabbit non-immune serum. The figure was generated based on the data represented in the Table 1 in “J. Nicoll, E. A. Gorbunov, S. A. Tarasov, O. I. Epstein, Subetta treatment increases adiponectin secretion by mature human adipocytes in vitro. Int J Endocrinol. 2013, Article ID 925874 (2013), doi:10.1155/2013/925874.”
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