FORMULATION FOR THE USE IN THE SIMULTANEOUS TREATMENT OF COCCIDIAL INFECTIONS AND IRON DEFICIENCIES

Abstract

The present invention relates to a formulation for use in the simultaneous treatment of coccidial infections and iron deficiencies containing a triazinone such as toltrazuril and a polynuclear iron(III) polysaccharide complex compound together with a specifically selected surfactant that has a hydrophilic-lipophilic-balance (HLB) value of ≥10.

Claims

1. A method for simultaneous treatment of a coccidial infection and an iron deficiency in a non-human animal, the method comprising administering to the non-human animal a formulation comprising toltrazuril or diclazuril, a polynuclear iron(III) polysaccharide complex compound and at least one surfactant having a HLB value of 10 or more.

2. The method according to claim 1, with the exception of the formulation is selected from the list consisting of a. a formulation, consisting of 3.5% toltrazuril, 17.8% of iron (as Gleptoferron), 0.3% of sorbitan monooleate, 10% diethylene glycol monoethyl ether, 0.1% of copolymer Polyoxyl 35—hydrogenated castor oil, and Water ad 100%; b. a formulation, consisting of 3.5% toltrazuril, 19.4% of iron (as Gleptoferron), 0.3% of sorbitan monooleate, 0.1% of copolymer Polyoxyl 35—hydrogenated castor oil, and Water ad 100%; c. a formulation, consisting of 3.5% toltrazuril, 19.3% of iron (as Gleptoferron), 0.3% of Propylene Glycol Monolaurate, 0.1% of copolymer Polyoxyl 35—hydrogenated castor oil, 1% Sodium Chloride, and Water ad 100%; and d. a formulation, consisting of 3.5% toltrazuril, 19.4% of iron (as Gleptoferron), 0.3% of sorbitan monooleate, 0.1% of copolymer Polyoxyl 35—hydrogenated castor oil, and Water ad 100%.

3. The method according to claim 1, with the exception of the formulation is selected from the list consisting of a. a formulation, consisting of 4.2% toltrazuril, 13.5% of iron (as Gleptoferron), 0.1% Sodium Docusate, 0.1% silicone emulsion, 1.5% sodium chloride, and Water ad 100%. b. a formulation, consisting of 4.2% toltrazuril, 13.5% of iron (as Gleptoferron), 0.1% Sodium Docusate, 5% diethylene glycol monoeethyl ether, 0.1% silicone emulsion, 1% sodium chloride, and Water ad 100%. c. a formulation, consisting of 4.2% toltrazuril, 13.5% of iron (as Gleptoferron), 0.6% Phenole, 0.2% Sodium Docusate, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicone dioxide, 1% povidone, 0.5% sodium chloride, and Water ad 100%. d. a formulation, consisting of 4.2% toltrazuril, 13.5% of iron (as Gleptoferron), 0.6% Phenole, 0.2% Sodium Docusate, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicone dioxide, 1% povidone, 1% sodium chloride, and Water ad 100%. e. a formulation, consisting of 4.2% toltrazuril, 13.5% of iron (as Gleptoferron), 0.6% Phenole, 0.1% Sodium Docusate, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicone dioxide, 1% povidone, 1% sodium chloride, and Water ad 100%; and f. a formulation, consisting of 2.9% toltrazuril, 18.8% of iron (as Gleptoferron), 0.6% Phenole, 0.1% Sodium Docusate, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicone dioxide, 1% povidone, 1% sodium chloride, and Water ad 100%.

4. The method according to claim 1, with the exception of the formulation is selected from the list consisting of a. a formulation, consisting of 2.9% toltrazuril, 18.8% of iron (as Gleptoferron), 0.3% of sorbitan monooleate, 0.1% of copolymer 15 Polyoxyl 35—hydrogenated castor oil, 1.5% of sodium chloride and water; b. a formulation, consisting of 4.2% toltrazuril, 13.5% of iron (as Gleptoferron), 0.3% of sorbitan monooleate, 0.1% of copolymer Polyoxyl 35—hydrogenated castor oil, 1.5% of sodium chloride and water; and c. a formulation, consisting of 4.2% toltrazuril, 13.5% of iron (as Gleptoferron), 1% of sodium chloride, 1% of polyvinylpyrrolidone, 0.6% of phenol, 0.5% of colloidal silicon dioxide, 0.1% of sodium docusate, 0.1% of simethicon emulsion and water

5. (canceled)

6. The method according to claim 1, except for those formulations comprising Copolymer Polyoxyl 35—hydrogenated castor oil or sodium docusate as a surfactant, when toltrazuril is used.

7. The method according to claim 1, wherein the at least one surfactant having a HLB value of 10 or more is selected from the list of Sodium Docusate, Polysorbates, Copolymer Polyoxyl 35—hydrogenated castor oil, Pluronic F-68 and Vitamin E TPGS (HLB 13.2).

8. The method according to claim 1, wherein the at least one surfactant having a HLB value of 10 or more is selected from the list of Polysorbate 20 and Polysorbate 80.

9. The method according to claim 8, wherein at least two surfactants with a HLB value of ≥10 are present in the formulation.

10. The method according to claim 9, wherein the at least two surfactants are Polysorbate 20 and Polysorbate 80.

11. The method according to claim 1, wherein the formulation is a suspension.

12. The method according to claim 1, wherein the formulation is administered by oral application.

13. The method according to claim 1, wherein the formulation is administered by parenteral application.

14. The method according to claim 13 wherein the formulation is administered by injection application.

Description

EXAMPLES

[0177] Preparation of formulations used in examples 1 to 4.

[0178] Formulations of toltrazuril used in the following examples 1 to 4 are produced by weighing the necessary amount of toltrazuril into a beaker and dissolving it in about half of the amount of the lead solvent.

[0179] Next, if applicable, the specified amount of the co-solvent ethanol and the surfactants are added. The formulation is stirred until a clear solution is obtained. The formulation is transferred into a volumetric flask, tempered to 20° C. and filled with the lead solvent up the calibration mark (q.s to the desired volume). The formulations are sterile filtered into injection vials and closed with rubber stoppers and crimped caps.

[0180] List of surfactants/dispersants used:

TABLE-US-00001 (Trade) name(s) Chemical classification or Pharm. Eur. Name Kollidone ™ 17PF Polyvinylpyrrolidone Cremophor ™ EL/ Polyoxyl 35-hydrogenated castor oil (USP/NF) Kolliphor ™ EL Tween ™ 80 Polysorbate 80 (Pharm. Eur.), Polyoxyethylen(20)-sorbitan-monooleat (IUPAC) Pluronic ™ F68 Poloxamer 188 (Pharm. Eur.), Ethylene Oxide/Propylene Oxide Block Copolymer, Poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) Lipoid ™ S100 (purified) phosphatidylcholine from fatfree soybean lecithin Vitamin E TPGS Vitamine E-Tocopheryl-polyethyleneglycole- succinate

[0181] Administration of the formulations used in examples 1 to 4 and evaluation:

[0182] Aliquots of 1 ml/kg of the formulations were injected subcutaneously to female Wistar rats. Blood samples are collected at different time intervals and the plasma samples analyzed with regard to the concentration of toltrazuril in the plasma. From those data the AUC and Cmax values are determined according to standard methods, known to clinicians.

Example 1

[0183] Formulations of toltrazuril in the solvent dimethylsulfoxide (DMSO) are prepared according to Table 1 with the compositions of the formulations given in % mass per Volume (% M/V). The content of DMSO is meant as q.s. ad 100. In Table 2 the pharmacokinetic AUC and Cmax data and the change in % compared to the reference formulation “ref” that does not comprise a surfactant are shown. [Note: “hr”=hours].

TABLE-US-00002 TABLE 1 Formulations of Toltrazuril in DMSO Function Ingredient HLB ref 1 2 3 4 5 API Toltrazuril n.a. 1 Dispersant Kollidone ™ 17PF n.a. — — — — — 7.5 Surfactant Cremophor ™ EL 12-14 — 10 — — — — Surfactant Tween ™ 80 15 — — 10 — — — Surfactant Vitamin E TPGS 13.2 — — — 10 — — Surfactant Pluronic ™ F68 29 — — — — 5 — Solvent Ethanol n.a. 10 Solvent DMSO n.a. 89 79 79 79 84 81.5

TABLE-US-00003 TABLE 2 Pharmacokinetic data of formulations according to Example 1 PK data Ref 1 2 3 4 5 AUC 778367 988485 1010307 922276 967051 810136 (hr*μg/L) % Change — +27 +30 +18 +24 +4 of AUC Cmax  11632 15920 14643 13887 13424 12638 (μg/L] % Change — +37 +26 +19 +15 +9 of Cmax

[0184] The above data clearly show the positive effect of the addition of surfactants with a HLB≥10, while the effect of the dispersant Kollidone™ 17PF which is not an amphiphilic compound and therefore intrinsically does not have a calculable HLB value, is considerably less.

Example 2

[0185] Formulations of toltrazuril in the solvent glycerol formal (4-Hydroxymethyl-1,3-dioxolan) are prepared according to Table 3 Table lwith the compositions of the formulations given in % mass per Volume (% MN). The content of glycerol formal is meant as q.s. ad 100. In Table 2 Table 4 the pharmacokinetic AUC and Cmax data and the change in % compared to the reference formulation “ref” that does not comprise a surfactant are shown. [Note: “hr”=hours]

TABLE-US-00004 TABLE 3 Formulations of Toltrazuril in glycerol formal Function Ingredient HLB ref 1 2 3 API Toltrazuril n.a. 1 Surfactant Cremophor ™ EL 12-14 — 10 — — Surfactant Tween ™ 80 15 — — — 10 Surfactant Vitamin E TPGS 13.2 — — 10 — Solvent Glycerol formal n.a. 99 89 89 89 (q.s. ad 100)

TABLE-US-00005 TABLE 4 Pharmacokinetic data of formulations according to Example 2 PK data ref 1 2 3 AUC 928370 1032468 969224 980867 (hr*μg/L) % Change — +11 +4 +6 of AUC Cmax  12785 14563 14825 16103 (μg/L] % Change — +14 +16 +26 of Cmax

[0186] The above data clearly show the positive effect of the addition of surfactants with a HLB≥10, which is less pronounced in the solvent glycerol formal, but still significant.

Example 3 (Comparative Example)

[0187] Formulations of toltrazuril in the solvent glycerol formal (4-Hydroxymethyl-L3-dioxolan) and 10% ethanol are prepared according to Table 5 Table lwith the compositions of the formulations given in % mass per Volume (% M/V). The content of glycerol formal and 10% ethanol is meant as q.s. ad 100. In Table 6 Table 2 the pharmacokinetic AUC and Cmax data and the change in % compared to the reference formulation “ref” that does not comprise a surfactant are shown. [Note: “hr”=hours]

TABLE-US-00006 TABLE 5 Formulations of Toltrazuril in glycerol formal and 10% ethanol Function Ingredient HLB ref 1 API Toltrazuril n.a. 1 Surfactant Lipoid ™ S100 <10 — 10 Solvent Ethanol n.a. 10 Solvent Glycerol formal n.a. 89 79 (q.s. ad 100)

TABLE-US-00007 TABLE 6 Pharmacokinetic data of formulations according to Example 3 PK data ref 1 AUC 1012525 732434 (hr*μg/L) % Change — −28 of AUC Cmax  15452 12220 (μg/L] % Change — −21 of Cmax

[0188] The above data show that the addition of the surfactant Lipoid S100 which has a HLB of <10 results in a negative effect on the pharmacokinetic parameters.

Example 4

[0189] Formulations of toltrazuril in the solvent 2-pyrrolidone and 20% ethanol are prepared according to Table 7 Table 1 with the compositions of the formulations given in % mass per Volume (% MN). The content of 2-pyrrolidone and 20% ethanol is meant as q.s. ad 100. In Table 8 the pharmacokinetic AUC and Cmax data and the change in % compared to the reference formulation “ref” that does not comprise a surfactant are shown. [Note: “hr”=hours]

TABLE-US-00008 TABLE 7 Formulations of Toltrazuril in 2-pyrrolidon and 20% ethanol Function Ingredient HLB ref 1 2 3 4 API Toltrazuril n.a. 1 Surfactant Tween ™ 80 15 — 10 — — — Surfactant Cremophor ™ EL 12-14 — — 10 — — Surfactant Vitamin E TPGS 13.2 — — — 10 — Surfactant Pluronic ™ F68 29 — — — — 5 Solvent Ethanol n.a. 20 Solvent 2-Pyrrolidone n.a. 79 69 69 69 74

TABLE-US-00009 TABLE 8 Pharmacokinetic data of formulations according to Example 4 PK data ref 1 2 3 4 AUC 498930 676594 659221 515567 555691 (hr*μg/L) % Change — +36 +32 +3 +11 of AUC Cmax  12396 16147 15545 13320 14473 (μg/L) % Change — +30 +25 +7 +17 of Cmax

[0190] The above data clearly show the positive effect of the addition of surfactants with a HLB≥10 also in 2-pyrrolidone and 20% ethanol as a solvent.

Example 5

[0191] Examples 1 to 4 have shown the individual, positive effect of the addition of a surfactant of a HLB≥10. To show the combined effect of said surfactants with the addition of a polynuclear iron(III) polysaccharide complex, formulations as outlined in Table 9 were prepared in analogy to those disclosed in EP 2 164 496 A1.

TABLE-US-00010 TABLE 9 Formulations of Toltrazuril in water with and w/o Iron Function Ingredient HLB Ref. 1 API Toltrazuril n.a. 5 5  API Fe(III)Dextran n.a. — 22.8 Preservative Sodium n.a. 0.2 Proprionate Preservative Sodium Benzoate n.a. 0.2 Surfactant Sodium Docusate 10 0.25 Defoamer Simethicone n.a. 0.05 Emulsion USP 30% Antifreezing Propylene n.a. 10.5 Glycole pH-Regulator Citric Acid n.a. 0.6-1.03 (ad pH ~4.5)   Thickener Bentonit/ n.a. 0.15-0.65 (ad ~150 mPa .Math. s) Xanthan gum Solvent Water n.a. ad 100

[0192] A total of 21 piglets born to 2 sows were randomly allocated to 2 study groups of 10 and 11 piglets each. Both formulations were administered in a single oral bolus to 3 day old piglets of the respective groups.

[0193] Frequent blood samplings (n=22 samples per piglet) were performed after treatment following a pre-determined schedule over a total period of 84 days, including a pre-treatment sample.

[0194] Concentrations of the active substance toltrazuril in plasma were analyzed by turbulent flow chromatography/tandem mass spectrometry. The limit of quantitation was 10 μg/L.

[0195] From the samples taken, the PK data of Table 10 were calculated.

TABLE-US-00011 TABLE 10 Pharmacokinetic data of formulations according to Example 5 PK data ref 1 AUC 36.07 51.57 (day*mg/L) % Change — +42.9 of AUC Cmax  8.44 10.07 (mg/L) % Change — +19.3 of Cmax

[0196] The above data clearly show the positive combined effect of the addition of a surfactant with a HLB≥10 with a polynuclear iron(III) polysaccharide complex compound.

[0197] While examples 1 to 4 showed the positive effect of the surfactant with a HLB≥10, the data generated as to Table 10 show an even further increased improvement of the relevant pharmacokinetic parameters.

Example 6

[0198] Examples 1 to 4 have shown the individual, positive effect of the addition of a surfactant of a HLB>10. Example 5 has shown the positive combined effect of said surfactants with the addition of a polynuclear iron(III) polysaccharide complex in an oral application.

[0199] To prove that the overall combined effect is not subject to a specific route of administration, formulations as outlined in Table 11 were prepared to be administered via injection. The “ref1” item is the same as outlined above in Example 5 to be the “ref” item, meaning an oral formulation only comprising toltrazuril. The “ref2” item is identical to the test item 1 of this example, but misses the addition of active iron.

[0200] Formulations were produced in analogy to Example 5.

TABLE-US-00012 TABLE 11 Formulations of Toltrazuril in water with and w/o Iron Function Ingredient HLB Ref1. Ref2. 1 API Toltrazuril n.a. See 3 API Fe(III)Dextrangluco- n.a. Example — 18.2 heptonate 5 under Preservative Phenole n.a. “ref” 0.5 Surfactant Tween ™ 20 16.7 0.14 Surfactant Tween ™ 80 15 0.09 Salt Sodium Chloride n.a. 0.9 Solvent Water n.a. ad 100

[0201] In a parallel design 3 study groups of 6 piglets each were set-up. The animals were treated on their 3.sup.rd day of life with the respective items. The piglets of the group receiving the formulation as to refl. received (just as above in Example 5) a single oral bolus, while the piglets of the respective two other groups received a single injection of the two formulations on their 3.sup.rd day of life.

[0202] Frequent blood sampling was performed after each treatment at 0 h, 4 h, 8 h, 24 h, 32 h, 48 h, 72 h, 7 d, 14 d, 21 d and 28 d (n=11/animal). Concentrations of the active substance toltrazuril in plasma were analyzed by turbulent flow chromatography/tandem mass spectrometry. The limit of quantitation was 25 μg/L for each analyte.

[0203] From the samples taken, the PK data of Table 12 were calculated.

TABLE-US-00013 TABLE 12 Pharmacokinetic data of formulations according to Example 6 PK data Ref1 Ref2 1 AUC 897 1920 2224 (hr*mg/L) % Change +214 +248 of AUC

[0204] The above data clearly show the positive combined effect of the addition of a surfactant with a HLB≥10 with a polynuclear iron(III) polysaccharide complex compound regardless of route of administration, while the even higher HLB surfactants compared to that of Example 5 seems to further increase the positive effect on the pharmacokinetic properties achieved.

[0205] In summary of the above Examples 1 to 6 it can be established that surfactants of a HLB value of ≥10 substantially increase the already amended pharmacokinetic properties of a toltrazuril polynuclear iron(III) polysaccharide complex compound combination, that such effect is not bound to the formulation being a solution or dispersion of the actives and that such effect is neither subject of a specific route of application thereof.