SEMI-SOLID FORMULATION FOR VOLUNTARY ORAL ADMINISTRATION OF BIOACTIVE COMPOUNDS TO RODENTS

Abstract

A formulation for voluntary oral administration of bioactive compounds to rodents. The semidsolid formulation having at least one thickening agent, a digestible sweetener selected from sucralose and calcium saccharin, a diluent agent, wherein the diluent agent is a powder with a granulometry between 300 and 800 μm, obtained from finely grounded seeds, cereal grains or cereal grain based diet for rodents, at least one flavor masking agent and a bioactive compound. The semi-solid formulation promotes voluntary feeding in rodents regardless of the bioactive compounds contained therein and without disruption of the metabolic pathways, therefore the semi-solid formulation has shown to be ideal for use in the oral administration of drugs and bioactive compounds to rodents. Use of the semi-solid formulation in the production of toxicant baits for rodents is also envisioned.

Claims

1. A semi-solid pill formulation for the voluntary oral administration of bioactive compounds to rodents comprising: at least one thickening agent, wherein said thickening agent is selected from the group consisting of gelatin, agar, carrageenan, sodium alginate and pectin; a digestible sweetener, wherein the digestible sweetener is selected from the group consisting of sucralose and calcium saccharin; a diluent agent, wherein the diluent agent is a powder with a granulometry between 300 and 800 μm in a concentration between 10 and 30% m/v, wherein the powder is selected from finely grounded seeds, cereal grains or finely grounded cereal grain based diet for rodents; at least one flavor masking agent; a bioactive compound.

2. The semi-solid pill formulation according to claim 1, wherein the finely grounded seeds are selected from at least one of sunflower seeds and canary seeds.

3. The semi-solid pill formulation according to claim 1, wherein the finely grounded cereal grain based diet for rodents comprises at least one of dehulled soybean meal, wheat middlings, flaked corn, ground corn, fish meal, cane molasses, ground wheat, dried whey, soybean oil and brewers dried yeast.

4. The semi-solid pill formulation according to claim 1, wherein the semi-solid pill formulation further comprises a preservative.

5. The semi-solid pill formulation according to claim 1, wherein the thickening agent is gelatin in a concentration between 5 and 20% m/v.

6. The semi-solid pill formulation according to claim 1, wherein the thickening agent is agar in a concentration between 1 and 5% m/v.

7. The semi-solid pill formulation according to claim 1, wherein the thickening agent is carrageenan in a concentration between 0.02 and 3% m/v.

8. The semi-solid pill formulation according to claim 1, wherein the thickening agent is sodium alginate in a concentration between 0.7 and 2% m/v.

9. The semi-solid pill formulation according to claim 1, wherein the thickening agent is pectin in a concentration between 0.5 and 5% m/v.

10. The semi-solid pill formulation according to claim 1, wherein the non-digestible sweetener is sucralose in a concentration between 1 and 5% m/v.

11. The semi-solid pill formulation according to claim 1, wherein the non-digestible sweetener is calcium saccharin in a concentration between 0.1 and 0.25% m/v.

12. The semi-solid pill formulation according to claim 1, wherein the flavor masking agent is selected from vanilla, strawberry, blueberry and chocolate.

13. The semi-solid pill formulation described in claim 1, for use in the oral administration of drugs and bioactive compounds to rodents.

14. The semi-solid pill formulation described in claim 1, for use in toxicological screening methods of rodents.

15. A toxicant bait for rodents comprising the semi-solid pill formulation described in claim 1.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0067] For easier understanding of this application, figures are attached in the annex that represent the preferred forms of implementation which nevertheless are not intended to limit the technique disclosed herein.

[0068] FIG. 1 shows voluntary acceptance index of prior art formula (Zhang et al.) versus present invention formula (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M. (n=6-8 experimental group).

[0069] FIG. 2 shows Semi-solid vehicle turbidity over time. The transmitted light, assessed through spectrophotometry, was used as a readout of chemical stability under RT or 4° C. storage. Standard vehicle composition: gelatin porcine skin 20% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15% and preservatives [tartaric acid (E334) and sodium citrate (E331) added according to EMA dosage recommendations].

[0070] FIG. 3 illustrates the semi-solid pill administration setup.

[0071] FIG. 4 shows the semi-solid pill vehicle acceptance over time in healthy rodents (A: C57BL/6 mice, B: Wistar rats). Standard vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%; Low-protein vehicle composition: Agar 2% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, standard diet for rodents 15%). Data are presented as mean±S.E.M. (n=6-8 experimental group).

[0072] FIG. 5 shows the semi-solid pill vehicle acceptance carrying bioactive compounds over time in healthy C57BL/6 mice (Vehicle composition: described in Examples, Section 3.3). Data are presented as mean±S.E.M. (n=6-8 experimental group).

[0073] FIG. 6 shows the semi-solid pill vehicle acceptance carrying Warfarin (80 mg/Kg body weight), a rodenticide in healthy C57BL/6 mice (Vehicle composition: described in Examples, Section 3.3). Data are presented as mean±S.E.M. (n=6-8 experimental group).

[0074] FIG. 7 shows body weight evolution after chronic vehicle semi-solid pill administration (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M. (n=6-8 experimental group).

[0075] FIG. 8 shows behavioral tests to ascertain stress-related parameters after chronic vehicle semi-solid pill administration (35 consecutive days). Open field for exploration habits, Elevated Plus Maze for anxiety-like parameters, Y-Maze for willingness to explore new environments and Splash test for self-care and motivation. (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M. (n=11-12 experimental group).

[0076] FIG. 9 shows corticosterone plasmatic concentration 30 minutes following a single vehicle administration through forced gavage versus voluntary semi-solid vehicle consumption (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, standard diet for rodents 15%). Data are presented as mean±S.E.M. (n=5-7 experimental group. One-way ANOVA, ** P<0.01 versus control, ##P<0.01 versus vehicle).

[0077] FIG. 10 shows Ileum contractility after chronic vehicle semi-solid pill administration (35 consecutive days. Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M. (n=6-8 experimental group).

[0078] FIG. 11 illustrates Hematoxylin & eosin staining (10× magnification) of liver and kidney sections after chronic vehicle semi-solid pill administration ((35 consecutive days. Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%).

[0079] FIG. 12 shows metabolic profile and biochemical markers of liver and kidney function after chronic vehicle administration (35 consecutive days). HDL, high-density lipoprotein; LDL, low-density lipoprotein; ALT, alanine transaminase; AST, aspartate transaminase. (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M. (n=6-8 experimental group).

[0080] FIG. 13 shows semi-solid pill vehicle acceptance over time in an experimental mouse model of multiple sclerosis (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M. (One-way ANOVA, *** P<0.001 versus control, n=6-8 experimental group).

[0081] FIG. 14 shows Body weight evolution after chronic semi-solid pill administration in an experimental mouse model of multiple sclerosis (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M.

[0082] FIG. 15 illustrates plasmatic sitagliptin concentration (HPLC/UV) following sitagliptin administration (50 mg/Kg, once) through gavage or voluntary semi-solid vehicle consumption Data are presented as mean±S.E.M. (n=5-6 experimental group). (Vehicle composition: gelatin porcine skin 10% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%).

[0083] FIG. 16 shows In vivo drug efficacy after sitagliptin administration (50 mg/Kg, once) through gavage or through semi-solid pill. Plasmatic DPPIV activity was measured to assess sitagliptin pharmacological action (Vehicle composition: gelatin porcine skin 2% m/v, sucralose 2%, strawberry flavoring essence 8% v/v, cereal grain based standard diet for rodents 15%). Data are presented as mean±S.E.M. (One-way ANOVA, **** P<0.0001 versus control, n=6-8 experimental group).

DESCRIPTION OF THE EMBODIMENTS

[0084] Now, preferred embodiments of the present application will be described in detail. However, they are not intended to limit the scope of this application.

[0085] It is disclosed herein a semi-solid pill formulation for the voluntary oral administration of bioactive compounds to rodents, wherein said formulation comprises: [0086] at least one thickening agent, wherein said thickening agent is selected from gelatin, agar, carrageenan, sodium alginate and pectin; [0087] a digestible sweetener, wherein the digestible sweetener is selected from sucralose and calcium saccharin; [0088] a diluent agent, wherein the diluent agent is a powder with a granulometry between 300 and 800 μm, obtained from finely grounded seeds, cereal grains or cereal grain based diet for rodents, in a concentration between 10 and 30% m/v; [0089] at least one flavor masking agent; [0090] a bioactive compound.

[0091] In one embodiment, the finely grounded seeds are selected from at least one of sunflower and canary.

[0092] In one embodiment, the finely grounded cereal grain based diet for rodents comprise at least one of dehulled soybean meal, wheat middlings, flaked corn, ground corn, fish meal, cane molasses, ground wheat, dried whey, soybean oil and brewers dried yeast.

[0093] In one embodiment, the formulation further comprises a preservative.

[0094] In one embodiment, the thickening agent is gelatin in a concentration between 5 and 20% m/v.

[0095] In another embodiment, the thickening agent is agar in a concentration between 1 and 5% m/v.

[0096] In another embodiment, the thickening agent is carrageenan in a concentration between 0.02 and 3% m/v.

[0097] In another embodiment, the thickening agent is sodium alginate in a concentration between 0.7 and 2% m/v.

[0098] In another embodiment, the thickening agent is pectin in a concentration between 0.5 and 5% m/v.

[0099] In one embodiment, the non-digestible sweetener is sucralose in a concentration between 1 and 5% m/v.

[0100] In another embodiment, the non-digestible sweetener is calcium saccharin in a concentration between 0.1 and 0.25% m/v.

[0101] In one embodiment, the flavor masking agent is selected from vanilla, strawberry, blueberry and chocolate.

[0102] In one embodiment, the formulation of the present patent application is for use in the oral administration of drugs and bioactive compounds to rodents.

[0103] In another embodiment, the formulation of the present patent application is used in toxicological screening methods of rodents.

[0104] In another embodiment, the formulation of the present patent application is used in the production of toxicant baits for rodents.

[0105] This description is of course not in any way restricted to the forms of implementation presented herein and any person with an average knowledge of the area can provide many possibilities for modification thereof without departing from the general idea as defined by the claims. The preferred forms of implementation described above can obviously be combined with each other. The following claims further define the preferred forms of implementation.

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