SOOTHING PRO-PHEROMONAL COMPOSITION FOR MAMMALS

Abstract

The invention relates to a compound of general formula (I) (in configuration Z or E) and to pro-pheromonal compositions and formulations comprising said compound, in addition to the uses thereof for soothing purposes for non-human mammals such as sheep, pigs, sheep, cattle, felines, equines and canines.

##STR00001##

Claims

1. A compound of the general formula (1): ##STR00009## wherein n is equal to 0 or 1 and R represents a saturated or unsaturated linear alkyl group having 2 to 30 carbon atoms.

2. The compound as claimed in claim 1, characterized in that n=0 and R is a saturated or unsaturated linear alkyl group having 9 to 30 carbon atoms, more particularly 12 to 30 carbon atoms.

3. The compound as claimed in claim 1, characterized in that n=1 and R is a saturated or unsaturated linear alkyl group having 2 to 30 carbon atoms, more particularly 8 to 22 carbon atoms.

4. The compound as claimed in one of the preceding claims for use as a veterinary medicinal product for treating anxiety, stress or aggressiveness in a non-human mammal.

5. The compound as claimed in claim 4, characterized in that the non-human mammal is selected from the group consisting of pigs, sheep, horses, cattle, cats and dogs.

6. A veterinary composition comprising a compound (1) as claimed in one of claims 1 to 5 and at least one fatty acid or a derivative thereof.

7. The composition as claimed in claim 6, characterized in that the acid is a fatty acid having 6 to 30 carbon atoms or a derivative of such a fatty acid selected from esters, thioesters or amides.

8. The composition as claimed in claim 7, characterized in that the fatty acid is selected from the group consisting of lauric acid, linoleic acid, linolenic acid, palmitic acid, pentadecanoic acid, capric acid, oleic acid, myristic acid, palmitoleic acid, azelaic acid, pimelic acid and mixtures thereof.

9. The composition as claimed in one of claims 6 to 8, characterized in that the amount of the compound (1) is between 0.1% and 5% by weight of the composition.

10. The composition as claimed in one of claims 6 to 9, characterized in that the amount of fatty acid or fatty acid derivative is between 95% and 99.9% by weight of the composition.

11. A formulation comprising the composition as claimed in one of claims 6 to 10 and a solvent.

12. The formulation as claimed in claim 11, characterized in that the solvent is selected from the group consisting of water, alcohols, glycols, polyglycols, paraffin and mixtures thereof.

13. The formulation as claimed in claim 12, characterized in that the solvent is selected from the group consisting of water, ethanol, isopropanol, paraffin, dipropylene glycol methyl ether, propylene glycol propyl ether, tripropylene glycol methyl ether and mixtures thereof.

14. The composition as claimed in one of claims 6 to 10, for use as a veterinary medicinal product for treating anxiety, stress or aggressiveness in a non-human mammal.

15. The composition as claimed in claim 14, characterized in that the non-human mammal is selected from the group consisting of pigs, sheep, horses, cattle, cats and dogs.

16. The formulation as claimed in one of claims 11 to 13, for use as a veterinary medicinal product for treating anxiety, stress or aggressiveness in a non-human mammal.

17. The formulation as claimed in claim 16, characterized in that the non-human mammal is selected from the group consisting of pigs, sheep, horses, cattle, cats and dogs.

18. A device for administering to a non-human mammal the composition as claimed in one of claims 6 to 10 or the formulation as claimed in one of claims 11 to 13, characterized in that it is selected from the group consisting of: a vaporizer containing said composition or said formulation, an electric diffuser fitted with a wick and containing said composition or said formulation, an accessory designed to be carried by the non-human mammal and impregnated with said composition or said formulation.

19. A kit comprising the composition as claimed in any one of claims 6 to 10, the formulation as claimed in one of claims 11 to 13 and/or the device as claimed in claim 18, for decreasing stress, aggressiveness and/or anxiety in a non-human mammal.

Description

EXAMPLES

Example 1: Synthesis of a Compound According to the Invention Wherein R is C.SUB.22.H.SUB.43 .with n=0

[0075] Into a 250 mL three-neck round-bottom flask under nitrogen atmosphere is introduced cis-13-docosenol (15.0 g; 46.21 mmol) and triethylamine (9.6 mL; 69 mmol) dissolved in 50 mL of methyltetrahydrofuran. The reaction medium is cooled to 0° C. then mesyl chloride (3.9 mL; 50.80 mmol) in solution in 25 mL of methyltetrahydrofuran. The reaction medium is then allowed to return to room temperature and is stirred for 90 minutes. 100 mL of water is then introduced. The organic phase is then washed with 100 mL of aqueous HCl solution (0.1 N), 100 mL of water (emulsion), 100 mL of brine solution, dried over MgSO.sub.4, filtered and concentrated under vacuum. A colorless oil that crystallizes at room temperature is then obtained (16.1 g; 86%). The crude reaction product is used as such in the following condensation step with solketal (see .sup.1H NMR).

##STR00005##

[0076] Into a 500 mL three-neck round-bottom flask fitted with a Dean-Stark apparatus and under nitrogen atmosphere is introduced finely ground KOH suspended in toluene (110 mL). Solketal (4.3 mL; 34.7 mmol) and the mesylate derivative synthesized beforehand (15.9 g; 39.50 mmol) are introduced. The medium is then heated at reflux for 16 hours. The reaction is then quenched by addition of 200 mL of water then 100 mL of ethyl acetate is added so as to have two clear phases. The cloudy aqueous phase is then extracted again with 2×100 mL of ethyl acetate, washed with brine (100 mL), dried over MgSO.sub.4, filtered and concentrated under vacuum to lead to the expected product in the form of an orange oil (16.0 g; 93%).

##STR00006##

[0077] In a 500 mL single-neck round-bottom flask under nitrogen atmosphere, the dioxolane derivative (15.80 g; 36.0 mmol) is dissolved in a methanol/H.sub.2O mixture (158/15 mL). APTS (300 mg; 1.74 mmol) is then added to the reaction medium, which is heated at reflux for 3 hours. The medium is cooled then the methanol is evaporated under vacuum. The residue is then taken up in 100 mL of ethyl acetate, washed with saturated aqueous NaHCO.sub.3 solution, 100 mL of water, and brine (100 mL), dried over MgSO.sub.4, filtered and concentrated under vacuum to lead to an oil that crystallizes at room temperature. The product is crystallized in hexane (50 mL) to lead to a white solid (9.0 g; 63%).

##STR00007##

[0078] Into a 50 mL three-neck round-bottom flask under nitrogen atmosphere is introduced the C22:1 alkylglycerol derivative (7 g; 17.00 mmol) and pyridinium para-toluenesulfonate (43 mg; 0.17 mmol) in solution in toluene (90 mL). Tiglic aldehyde (11.5 mL; 0118 mmol) in solution in 10 mL of toluene is added to the reaction medium, which is then heated at reflux for 3 hours. The reaction medium is neutralized by addition of saturated NaHCO.sub.3 solution (50 mL). After decanting, the aqueous phase is extracted with ethyl acetate (50 mL).

[0079] The organic phases are then combined, washed with brine solution, dried over MgSO.sub.4, filtered and concentrated under vacuum to lead to a light-yellow oil (8.0 g). This oil is then purified by flash chromatography to lead to a colorless oil (6.0; 74%).

[0080] The table below presents the different alkylglycerols synthesized and sampled, as well as the operating conditions and the overall yields. The procedures used for the synthesis of C22:1 AKG appear in the experimental section.

##STR00008##

TABLE-US-00001 Electrophile Product 1 Yield R AKG Conditions leaving group formed (% mol.) C.sub.8H.sub.17 C.sub.8:0 A Iodine 1.sub.8 37 C.sub.10H.sub.21 C.sub.10:0 B Bromine 1.sub.10 68 C.sub.12H.sub.25 C.sub.12:0 B Bromine 1.sub.12 62 C.sub.14H.sub.29 C.sub.14:0 B Bromine 1.sub.14 65 C.sub.16H.sub.33 C.sub.16:0 B Mesylate 1.sub.16 66 C.sub.16H.sub.31 C.sub.16:1 C Mesylate 1.sub.16:1 66 C.sub.18H.sub.37 C.sub.18:0 B Mesylate 1.sub.18 65 C.sub.18H.sub.35 C.sub.18:1 C Mesylate 1.sub.18:1 66 C.sub.18H.sub.35 C.sub.18:2 C Mesylate 1.sub.18:2 64 C.sub.20H.sub.41 C.sub.20:0 B Bromine 1.sub.20 71 C.sub.22H.sub.43 C.sub.22:1 C Mesylate 1.sub.22:1 50

Conditions A: 1) Solketal/KOH/RI/DMSO/50° C. 2) APTS/MeOH/H.SUB.2.O.

Conditions B: 1) Solketal/KOH/RBr/Toluene/DS/2) APTS/MeOH/H.SUB.2.O.

[0081] Conditions C: 1) CH.sub.2Cl.sub.2/Et.sub.3N/MsCl 2) Solketal/KOH/ROMs/Toluene/DS/A 3) APTS/MeOH/H.sub.2O.

GC Method Used:

Equipment: GC, Hewlett Packard 5890 Series II

Detector FID

Column: HP5 30 m, 0.53 mm, 0.88 μm

Pressure: 11 psi

[0082] Injection volume: 1 μL
Sample preparation: 4 mg/mL in AcOEt
Injector temperature: 270° C.
Detector temperature: 280° C.

Oven:

[0083]

TABLE-US-00002 Initial temperature 170° C. Initial time 3 min Gradient 5° C./min Final temperature 260° C. Final time 10 min Run 31 min

Example 2: Preparation of Soothing Active Formulations

[0084] The soothing formulations are prepared in a precise order since the principle of the invention consists in combining a compound (1) with a fatty acid catalyst of its degradation. The method thus consists in introducing into a stirred-reactor type mixer the formulation solvent, the fatty acid or the mixture of fatty acids. Stirring is maintained for 1 hour, after which the medium is perfectly homogeneous, then the compound (1) is added and stirring is maintained for another hour.

[0085] The following table lists the mixtures that were prepared

TABLE-US-00003 Amount of Amount of Amount of Solvent solvent fatty acid Compound compound Example (v/v) (g) Fatty acids (g) (1) (1) (g) 2a Ethanol/water 85 Linoleic/ 12 1.sub.18 3 90/10 Pentadecanoic 95:5 2b Isopropanol 85 Linoleic/ 12 1.sub.18 3 Pentadecanoic 95:5 2c Isopar V 90 Linoleic 9.7 1.sub.22:1 0.3 paraffin 2d Isopar V 95 Linoleic 4.85 1.sub.22:1 0.15 paraffin 2e Isopar V 98 Linoleic 1.94 1.sub.22:1 0.06 paraffin 2f Dipropylene 99 Linoleic 0.9 1.sub.22:1 0.09 glycol methyl ether 2g Propylene 99 Linoleic 0.9 1.sub.22:1 0.09 glycol propyl ether 2h Propylene 99 Linoleic 0.9 1.sub.22:1 0.09 glycol methyl ether

Example 3: In Vitro Study of 2M2B Release

[0086] The solution prepared in Example 2b is used in a climatic chamber at 100° C. and 30° C. (conditions corresponding either to the temperature of a thermal diffuser or to room temperature). Samples are taken at regular intervals and the disappearance of compound 1.sub.22:1 and the appearance of C.sub.22:1 AKG is measured by gas chromatography. The results are presented in the following table (the percentages are expressed only taking into consideration integration relative to compound 1.sub.18 and of C.sub.18:0 AKG):

TABLE-US-00004 T = 100° C. 0 3 h 6 h 1 d 2 d 7 d 14 d 21 d 28 d % 1.sub.18 100 52 25 2 0 0 0 0 0 % C.sub.18:0 0 48 75 98 100 100 100 100 100 AKG T = 30° C. 0 3 h 8 h 1 d 2 d 7 d 14 d 21 d 28 d % 1.sub.18 100 100 100 100 99.5 99 99 % C.sub.18:0 0 0 0 0 0.5 1 1 AKG

[0087] This table shows that degradation of the product 1.sub.18 is well stimulated by high temperature (electric diffuser-type) but that in its diluted state at room temperature the solution is sufficiently stable to provide the treatment for one month (less than 5% loss over the period).

[0088] Likewise, we noted that the solutions of Examples 2f, 2 g and 2h have similar trans-2-methyl-2-butenal release kinetics and this despite a different kind of dilution solvent.

Example 4: Formulation for Thermal Diffuser and Release Curve

[0089] 50 mL of the formulations prepared in Examples 2c, 2d and 2e are used to fill three PET flasks fitted with a diffusion wick. Each flask is screwed onto an electric diffuser characterized in that the ceramic of which it is composed is heated to about 100° C. when the system is connected up.

[0090] The daily release of the formulation is then measured and the results are presented in the following table: Evaluated is the total loss of weight of the formulation reflecting the release of 2-methyl-2-butenal, the fatty acid and the combined solvent.

TABLE-US-00005 Days Formula 1 2 3 4 5 6 7 14 21 28 35 42 49 56 2c 1 0.5 0.3 0.3 0.2 0.2 0.1 0.1 0.05 0.05 — — — — 2d 0.9 1 0.8 0.6 0.4 0.2 0.2 0.15 0.15 0.15 0.15 0.15 0.15 0.15 2e 0.9 1.2 1.2 1.2 0.7 0.8 0.7 0.4 0.4 0.3 0.3  0.3  0.3  0.3 

[0091] The table expresses the daily diffusion in grams of each formulation

[0092] At day 35, the residual compositions in each flask are measured by liquid chromatography and compared with the initial compositions.

TABLE-US-00006 Composition after Formula Initial composition 35 d of diffusion 2c 90/9.7/0.3 88/11.6/0.1 2d 95/4.85/0.15 95/4.95/0.15 2e 98/1.94/0.06 98/1.94/0.06

[0093] The table expresses the content of each solution according to the ratio paraffin/linoleic acid/1.sub.22:1

[0094] Based on the preceding measurements, we can estimate that the amounts released over the 35-day period by molecules going into each composition are the following (in grams):

TABLE-US-00007 Formula 2c 2d 2e Linoleic acid 0.195 0.188 0.2 1.sub.22:1 0.05 0.0063 0.0063

[0095] These results show that the product 1.sub.22:1 and linoleic acid disappear from the formulation at flow rates that respect the orders of magnitude defined in the formulation.

[0096] By considering that at 100° C., all of product 1.sub.22:1 decomposes as was shown in Example 3, we can estimate the daily amounts of the various products released into the atmosphere according to the various formulae.

2% Formula (2e)

[0097]

TABLE-US-00008 Product Daily amount (mg) Linoleic acid 5.8 C.sub.22:1 AKG 0.14 2-Methyl-2-butenal 0.036

5% Formula (2d)

[0098]

TABLE-US-00009 Product Daily amount (g) Linoleic acid 5.1 C.sub.22:1 AKG 0.14 2-Methyl-2-butenal 0.036

10% Formula (2c)

[0099]

TABLE-US-00010 Product Daily amount (mg) Linoleic acid 5.5 C.sub.22:1 AKG 1.2 2-Methyl-2-butenal 0.32

Example 6: Study of the Soothing Effect on Pigs

[0100] The tests were carried out with electric diffusers like those described in Example 5.

[0101] The protocol is as follows:

[0102] One hundred 3-week-old piglets are distributed among 4 pigsties on 25 m.sup.2 grating (or 175 m.sup.3). The piglets are divided into populations of 25 called A, B, C, D. Population A is treated with a diffuser containing only Isopar V paraffin.

[0103] Population B is treated by a diffuser containing the formula of Example 2e (2%).

[0104] Population B is treated by a diffuser containing the formula of Example 2d (5%).

[0105] Population B is treated by a diffuser containing the formula of Example 2c (10%).

[0106] The tests are conducted for 28 days. The average weight of each population and the average number of visible wounds are measured each week.

[0107] The results are presented in the following tables:

TABLE-US-00011 Weight change as a function of piglet age (kg) A B C D Weight at 21 days 6.513 6.513 6.513 6.513 Weight at 28 days 7.081 7.075 7.078 7.051 Weight at 35 days 9.153 9.161 9.175 9.169 Weight at 42 days 11.815 12.012 12.02 12.07 Weight at 49 days 14.989 15.206 15.281 15.601

TABLE-US-00012 Daily average weight gains (kg) A B C D Week 1 0.08 0.08 0.08 0.08 Week 2 0.30 0.30 0.30 0.30 Week 3 0.38 0.41 0.41 0.41 Week 4 0.45 0.46 0.47 0.50

[0108] The number of bites over the four weeks of treatment is 10% to 12% lower for populations B, C and D compared with population A.

Example 7: Soothing Effect on Kittens

[0109] A 60 mL vaporizer is filled with the formulation of Example 2a. Another vaporizer containing a methanol/water mixture (90:10 v/v) is used as a placebo.

[0110] Six 1-month-old kittens from the same litter are separated from their mother and divided into two groups of three (2 males and 1 female in each group). Group A is placed in a basket onto which formulation 2a is vaporized every 12 hours. Group B is placed in a basket vaporized every 12 hours with the placebo.

[0111] In the first week, the population A kittens are calm and sleep against one another 16 hours per day on average whereas the population B kittens sleep at most 14 hours and at the end of the third day regularly fight.

[0112] The behaviors of the two populations converge the second week.

[0113] This example shows that formulation A by vaporizer improves the kittens' adaptation to weaning.