N-CARBAMOYLPUTRESCINE TO ENHANCE MUSCLE PROTEIN SYNTHESIS

Abstract

The present invention relates to the field of food supplement and medicament supporting muscle metabolism, more precisely muscle protein synthesis. In particular, the invention relates to a composition comprising N-carbamoylputrescine (NCP). The invention further relates to the non-therapeutic use of N-carbamoylputrescine (NCP) to enhance muscle protein synthesis in a subject. Moreover, the invention also relates to N-carbamoylputrescine for its use as a medicament.

Claims

1. An edible product comprising a pharmaceutically acceptable salt of N-carbamoylputrescine of formula (I) ##STR00002##

2. An edible product according to claim 1, comprising an enteric vehicle.

3. An edible product according to claim 1, wherein said edible product is a food product or a beverage product.

4. An edible product according to claim 1, further comprising L-amino-acids, polyamines, carbohydrate compounds, or combinations thereof.

5. An edible product according to claim 4, wherein the L-amino-acid is histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, alanine, arginine, citrulline, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, ornithine, proline, serine, tyrosine, argininosuccinate, or decarboxylated forms thereof and deaminated forms thereof.

6. An edible product according to claim 4, wherein the polyamine is putrescine, spermine, spermidine, or agmatine.

7. An edible product according to claim 4, wherein the carbohydrate compound is glucose, fructose, galactose, or polysaccharides thereof.

8. A topical cosmetic composition comprising N-carbamoylputrescine and/or one of its salts, and a cosmetically acceptable excipient.

9-12. (canceled)

13. A composition comprising a pharmaceutically acceptable salt of N-carbamoylputrescine.

14-15. (canceled)

16. An edible product according to claim 2, wherein said edible product is a food product or a beverage product.

17. An edible product according to claim 1, wherein said edible product is a medicament.

18. An edible product according to claim 2, wherein said edible product is a medicament.

19. The topical cosmetic composition of claim 8, said composition comprising a pharmaceutically acceptable salt of N-carbamoylputrescine.

20. The topical cosmetic composition of claim 8, wherein said topical cosmetic composition is a powder, aerosol, plaster lotion, liniment, solution, emulsion, suspension, ointment, cream, paste or gel.

21. The composition of claim 13, further comprising a pharmaceutically acceptable carrier.

Description

BRIEF DESCRIPTION OF FIGURES

[0079] FIG. 1: Effect of administration of NCP on plasma urea, glutamine and ornithine in rats.

[0080] Plasma concentrations of urea, glutamine and ornithine were measured in rats untreated (control) (white bars) and rats whose diet consisted of 5 mg/day of NCP (gray bars) for 10 days (thus corresponding to 50 mg/kg of NCP (black bars) The results (meanSD) are expressed in mmol/l urea and pmol/l amino acids * P<0.05.

EXAMPLES

Reagents

[0081] Polyamines and other reagents were from Sigma-Aldrich (Lisle d'Abeau Chesnes, France).

[0082] N-carbamoylputrescine (NCP; Cinoberine) was synthetized as described in Ramani et al. (Anal. Biochem., 2012, 423(1):54-60)

Animal

[0083] Eight-week old Sprague-Dawley rats (Charles Rivers, Lyon, France) were used. They were placed in individual cages and acclimatized for two weeks to our animal facility. During this period they received standard rodent chow (A04, UAR, Villemoisson-sur-Orge, France) and water ad libitum.

[0084] Animal care and experimentation complied with French and European Community regulations for animal care and experimentation (Official Journal of the European Community 110 L 358, 18 Dec. 1986). The study protocol has been approved by the Regional ethic committee of Ile-de-France.

Animal Experimentation

[0085] After acclimatization twenty eight-month old rats were randomized into three groups in order to receive for two weeks a standard diet either alone (n=6; control group) or supplemented with NCP either 5 mg/kg/d (n=7, NCP5 group) or 50 mg/kg/d (n=7; NCP50 group). Their weight, behaviour and mortality were monitored throughout the feeding period.

[0086] At the end of the feeding period, the rats, in the fasted state, were anesthetized by isoflurane inhalation and euthanatized by decapitation.

[0087] Mixed blood was collected onto heparinized tubes and rapidly centrifuged. The liver was immediately removed and weighed, and a sample was cut off, frozen in liquid nitrogen, and stored at 80 C. until analysis. For the jejunum and ileum, the intestine was washed with cold NaCl (0.9%) and reverted. Thereafter the intestinal mucosa was scraped, rapidly frozen in liquid nitrogen, and stored at 80 C. until analysis. Two muscles of the hindlimb, tibialis (rich in type II fibres) and soleus (rich in type I fibres), and the right kidney were rapidly removed, weighed, frozen in liquid nitrogen, and stored at 80 C. until analysis. Body composition was assessed by dissection and lean mass (carcass), visceral fat mass, sub-cutaneous fat mass (subcutaneous fat and skin) and mass of the viscera were determined.

[0088] Biological parameters studied were: [0089] in tissues: NCP, protein and amino acid contents [0090] in plasma: amino acids, liver function tests (plasma aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and bilirubin), muscle enzymes (plasma creatine kinase, and lactate dehydrogenase), plasma electrolytes (sodium, potassium, chloride, bicarbonates), total protein, urea, creatinine, calcium, phosphate, magnesium, uric acid, glucose, cholesterol and triglycerides.

Results

General Effects of N-Carbamoyl Putrescine

[0091] No rats showed abnormal general behavior, and there was no mortality in the 3 groups during the study.

[0092] All animals achieve normal weight gain and there was no difference between rats receiving NCP compared to controls (Table 1).

[0093] Analysis of body composition showed no difference between the groups. Lean mass, fat mass, either total, abdominal or cutaneous, mass of the viscera, and liver mass were similar in the three groups (data not shown).

TABLE-US-00001 TABLE 1 Influence of NCP on rat weight. Results are presented as mean SD Control NCP 5 mg/kg NCP 50 mg/kg (n = 6) (n = 7) (n = 7) Initial weight (g) 263.3 11.4 264.9 16.sup. 264.9 8.2 Final weight (g) 314.2 10.7 313.1 18.1 316.1 13.9 Weight gain (g) 50.8 7.9 48.1 8.1 51.2 8.7 Daily weight gain (g/d) 3.63 0.57 3.44 0.58 3.65 0.62

Biological Effects of N-Carbamoylputrescine

[0094] No difference was observed between the three groups in terms of plasma electrolytes levels (sodium, potassium, chloride, bicarbonates, calcium, phosphate, magnesium), in hepatic and muscular enzymes (aminotransferases, creatine kinase, lactate dehydrogenase, alkaline phosphatase), in creatinine, glucose, cholesterol, triglycerides and uric acid levels (data not shown). Plasma urea was higher in NCP50 group vs NCP5 (FIG. 1); however no difference was shown between the two NCP-supplemented groups compared to control rats.

[0095] Analysis of amino acid levels showed that plasma glutamine levels were significantly lower in the NCP50 group than in controls (FIG. 1) and there was a trend for a NCP dose-effect relationship (p=0.06). Plasma ornithine showed a similar dose-effect relationship (p=0.055) (FIG. 1). No differences were observed for other amino acids. There was no correlation between plasma urea and plasma glutamine or ornithine.

[0096] While liver, kidney, tibialis, and jejunal and ileal mucosa protein contents were similar between the three groups, protein level in soleus was higher in NCP5 vs control group (Control: 12.71.8 g/100 g, NCP5: 14.50.7 g/100 g, NCP50: 14.00.8 g/100 g; p=0.018 NCP5 vs control).

[0097] Also, in soleus muscle, while the concentration of most amino acids were similar between the 3 groups, the concentration of Glutamine, alanine and histidine significantly increased with NCP administration and there was a positive correlation for glutamine and alanine with the dose of NCP (p=0.015 and p=0.013 respectively)(Table 2)

TABLE-US-00002 TABLE 2 Soleus amino acid content. Results presented as mean SD; ANOVA and plsd Fisher a b:p < 0 Amino acid Control NCP 5 mg/kg NCP 50 mg/kg (mol/g) (n = 6) (1) (n = 7) (2) (n = 6) (3) Taurine 22972 3178 23882 2585 25223 1987 Aspartate 3889 612 4195 703 4742 1443 Threonine 481 75 512 109 571 67 Serine 1745 158 1933 384 1964 248 Asparagine 447 57 519 106 538 124 Glutamate 3757 474 3753 684 4074 390 Glutamine .sup.6858 1008.sup.a .sup.7605 1016.sup.ab .sup.8547 988.sup.b Glycine 1823 190 1932 516 2145 153 Alanine 1539 238.sup.a .sup.1735 385.sup.ab .sup.2025 182.sup.b Citrulline 321 33 351 53 379 65 Valine 172 38 182 22 194 24 Isoleucine 79 25 83 17 91 22 Leucine 97 34 103 23 113 32 Tyrosine 114 19 106 15 126 21 Phenylalanine 77 14 75 19 77 11 Ornithine 73 21 75 12 74 15 Histidine 406 70.sup.a 415 50.sup.a 494 6.sup.8b Lysine 877 249 933 222 1009 168 Arginine 318 71 381 115 395 62 Proline 207 39 196 42 199 26