阅读说明：本技术 用于同时治疗球虫感染和铁缺乏症的制剂 (Formulation for the simultaneous treatment of coccidial infections and iron deficiency ) 是由 S·霍夫曼 K·伯恩 M·维辛 于 2019-05-29 设计创作，主要内容包括：本发明涉及一种用于同时治疗球虫感染和铁缺乏症的制剂,其含有三嗪酮诸如托曲珠利和多核铁(III)多糖络合物以及具有≥10的亲水亲脂平衡(HLB)值的特别选择的表面活性剂。(The present invention relates to a formulation for the simultaneous treatment of coccidial infections and iron deficiency, containing triazinones such as toltrazuril and polynuclear iron (III) polysaccharide complexes and a specifically selected surfactant having a hydrophilic-lipophilic balance (HLB) value of 10 or more.)

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

2. The formulation for use according to claim 1, excluding formulations selected from the list consisting of:

a. a formulation consisting of: 3.5% toltrazuril, 17.8% iron (as iron glucoheptonate), 0.3% sorbitan monooleate, 10% diethylene glycol monoethyl ether, 0.1% copolymer polyethylene glycol 35-hydrogenated castor oil and water to 100%;

b. a formulation consisting of: 3.5% toltrazuril, 19.4% iron (as iron glucoheptonate), 0.3% sorbitan monooleate, 0.1% copolymer polyethylene glycol 35-hydrogenated castor oil and water to 100%;

c. a formulation consisting of: 3.5% toltrazuril, 19.3% iron (as iron glucoheptonate), 0.3% propylene glycol monolaurate, 0.1% copolymer polyethylene glycol 35-hydrogenated castor oil, 1% sodium chloride and water to 100%; and

d. a formulation consisting of: 3.5% toltrazuril, 19.4% iron (as iron glucoheptonate), 0.3% sorbitan monooleate, 0.1% copolymer polyethylene glycol 35-hydrogenated castor oil and water to 100%.

3. The formulation for use according to claim 1, excluding formulations selected from the list consisting of:

a. a formulation consisting of: 4.2% toltrazuril, 13.5% iron (as iron glucoheptonate), 0.1% sodium docusate, 0.1% silicone emulsion, 1.5% sodium chloride and water to make up to 100%;

b. a formulation consisting of: 4.2% toltrazuril, 13.5% iron (as iron glucoheptonate), 0.1% docusate sodium, 5% diethylene glycol monoethyl ether, 0.1% silicone emulsion, 1% sodium chloride and water to make up to 100%;

c. a formulation consisting of: 4.2% toltrazuril, 13.5% iron (as iron glucoheptonate), 0.6% phenol, 0.2% docusate sodium, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicon dioxide, 1% povidone, 0.5% sodium chloride and water to make up to 100%;

d. a formulation consisting of: 4.2% toltrazuril, 13.5% iron (as iron glucoheptonate), 0.6% phenol, 0.2% docusate sodium, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicon dioxide, 1% povidone, 1% sodium chloride and water to make up to 100%;

e. a formulation consisting of: 4.2% toltrazuril, 13.5% iron (as iron glucoheptonate), 0.6% phenol, 0.1% docusate sodium, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicon dioxide, 1% povidone, 1% sodium chloride and water to make up to 100%; and

f. a formulation consisting of: 2.9% toltrazuril, 18.8% iron (as iron glucoheptonate), 0.6% phenol, 0.1% docusate sodium, 0.1% simethicone emulsion USP 30%, 0.5% colloidal silicon dioxide, 1% povidone, 1% sodium chloride and water to make up to 100%.

4. The formulation for use according to claim 1, excluding formulations selected from the list consisting of:

a. a formulation consisting of: 2.9% toltrazuril, 18.8% iron (as iron glucoheptonate), 0.3% sorbitan monooleate, 0.1% copolymer 15 polyethylene glycol 35-hydrogenated castor oil, 1.5% sodium chloride and water;

b. a formulation consisting of: 4.2% toltrazuril, 13.5% iron (as iron glucoheptonate), 0.3% sorbitan monooleate, 0.1% copolymer polyethylene glycol 35-hydrogenated castor oil, 1.5% sodium chloride and water; and

c. a formulation consisting of: 4.2% toltrazuril, 13.5% iron (as iron glucoheptonate), 1% sodium chloride, 1% polyvinylpyrrolidone, 0.6% phenol, 0.5% colloidal silicon dioxide, 0.1% docusate sodium, 0.1% simethicone emulsion and water.

5. The formulation for use according to claim 1, excluding those selected from the list according to claims 2-4.

6. Formulations for use according to claim 1, when toltrazuril is used, those comprising the copolymer polyethylene glycol 35-hydrogenated castor oil or docusate sodium as surfactant are excluded.

7. The formulation for use according to any one of claims 1-5, wherein the at least one surfactant having an HLB value of 10 or higher is selected from the list of docusate sodium, polysorbate, copolymer polyethylene glycol 35-hydrogenated castor oil, pluronic F-68, and vitamin E TPGS (HLB 13.2).

8. The formulation for use according to any one of the preceding claims, wherein the at least one surfactant having an HLB value of 10 or higher is selected from the list of polysorbate 20 and polysorbate 80.

9. The formulation for use according to claim 8, wherein at least two surfactants having an HLB value of ≥ 10 are present in the formulation.

10. The formulation for use according to claim 9, wherein the at least two surfactants are polysorbate 20 and polysorbate 80.

11. The formulation for use according to any of the preceding claims, wherein the formulation used is a suspension.

12. The formulation for use according to any of the preceding claims, wherein use is made in oral administration.

13. The formulation for use according to any one of claims 1 to 11, wherein the use is carried out in parenteral administration.

14. The formulation for use according to claim 13, wherein the use is carried out in an injection administration.

Examples

Preparation of the formulations used in examples 1-4.

The formulations of toltrazuril used in examples 1-4 below were produced by weighing the necessary amount of toltrazuril into a beaker and dissolving it in about half the amount of the lead solvent.

Next, if applicable, the co-solvent ethanol and surfactant were added in the amounts indicated. The formulation was stirred until a clear solution was obtained. The formulation was transferred into volumetric flasks, tempered to 20 ℃, and filled to calibration mark (to desired volume) with lead solvent. The formulation was sterile filtered into an injection vial and closed with a rubber stopper and crimped crimp cap.

List of surfactants/dispersants used:

name of (business)	Chemical classification or European pharmacopoeia designation
		Kollidone™17PF	Polyvinylpyrrolidone
Clevelopv ™ EL- Kolliphor™ EL	Polyethylene glycol 35-hydrogenated Castor oil (USP/NF)
		Tween 80	Polysorbate 80 (European pharmacopoeia), polyoxyethylene (20) -sorbitan-monoolein Acid esters (IUPAC)
Pluronic ™ F68	Poloxamer 188 (european pharmacopoeia) ¸ ethylene oxide/propylene oxide block copolymer, poly (ethylene glycol) -poly (propylene glycol) -poly (ethylene glycol) block copolymers
		Lipoid™S100	(purified) phosphatidylcholine from defatted soy lecithin
Vitamin E TPGS	Vitamin E-tocopherol-polyethylene glycol-succinate

Administration and evaluation of the formulations used in examples 1-4:

an aliquot of 1ml/kg of the formulation was injected subcutaneously into female Wistar rats. Blood samples were collected at different time intervals and plasma samples were analyzed for toltrazuril concentration in plasma. AUC and Cmax values were determined from those data according to standard methods known to clinicians.

Example 1:

formulations of toltrazuril in the solvent dimethyl sulfoxide (DMSO) were prepared according to table 1, the composition of which is given in mass percent per volume (% M/V). The DMSO content is determined to be 100. In table 2, the pharmacokinetic AUC and Cmax data are shown as well as the change (expressed as a percentage) compared to the reference formulation "ref" without surfactant. [ note: "hr" = hour ].

TABLE 1 formulation of toltrazuril in DMSO

。

TABLE 2 pharmacokinetic data for the formulation according to example 1

。

The above data clearly show that the positive effect of the addition of a surfactant with an HLB > 10 is much less than that of the dispersant Kollidone 17PF (which is not an amphiphilic compound and therefore has essentially no calculable HLB value).

Example 2:

a formulation of toltrazuril in the solvent glycerol formal (4-hydroxymethyl-1, 3-dioxolane) was prepared according to table 3, the composition of which is given in mass percent per volume (% M/V). The content of glycerol formal is determined to be 100. In table 4, the pharmacokinetic AUC and Cmax data are shown as well as the change (expressed as a percentage) compared to the reference formulation "ref" without surfactant. [ note: "hr" = hour ].

Table 3: formulations of toltrazuril in glycerol formal

。

TABLE 4 pharmacokinetic data for the formulation according to example 2

。

The above data clearly demonstrate the positive effect of the addition of a surfactant with an HLB of > 10, which is less significant, but still significant, in the solvent glycerol formal.

Example 3 (comparative example):

a formulation of toltrazuril in the solvents glycerol formal (4-hydroxymethyl-1, 3-dioxolane) and 10% ethanol was prepared according to table 5, the composition of which is given in mass percent per volume (% M/V). The content of glycerol formal and 10% ethanol is defined as the volume to 100. In table 6, the pharmacokinetic AUC and Cmax data are shown as well as the change (expressed as a percentage) compared to the reference formulation "ref" without surfactant. [ note: "hr" = hour ].

TABLE 5 formulation of toltrazuril in glycerol formal and 10% ethanol

。

TABLE 6 pharmacokinetic data for the formulation according to example 3

。

The above data indicate that the addition of surfactant Lipoid S100 with HLB < 10 negatively affects pharmacokinetic parameters.

Example 4:

formulations of toltrazuril in the solvent 2-pyrrolidone and 20% ethanol were prepared according to table 7, the composition of which is given in mass percent per volume (% M/V). The contents of 2-pyrrolidone and 20% ethanol are specified to 100. Pharmacokinetic AUC and Cmax data and changes (expressed as percentages) compared to a reference formulation "ref" containing no surfactant are shown in table 8. [ note: "hr" = hour ].

TABLE 7 formulation of toltrazuril in 2-pyrrolidone and 20% ethanol

。

TABLE 8 pharmacokinetic data for the formulation according to example 4

。

The above data clearly demonstrate the positive effect of the addition of a surfactant with an HLB > 10 also in 2-pyrrolidone and 20% ethanol as solvents.

Example 5:

examples 1-4 have shown the sole positive effect of the addition of a surfactant with an HLB > 10. To show the combined effect of the surfactant with the addition of polynuclear iron (III) polysaccharide complex, the formulations outlined in table 9 were prepared similar to those disclosed in EP 2164496 a 1.

TABLE 9 formulation of toltrazuril in Water (with and without iron)

。

A total of 21 piglets born from 2 sows were randomly assigned to 2 study groups, 10 and 11 piglets per group. Both formulations were administered as a single oral bolus to each group of 3 day-old piglets.

Frequent blood sampling (n =22 samples/piglet) was performed on a predetermined schedule over a total period of 84 days after treatment, including samples before treatment.

The concentration of the active substance toltrazuril in the plasma was analyzed by means of turbulence chromatography/tandem mass spectrometry. The limit of quantitation was 10. mu.g/L.

PK data for table 10 were calculated from the collected samples.

TABLE 10 pharmacokinetic data of the formulation according to example 5

。

The above data clearly demonstrate the positive combination effect of the addition of a surfactant with an HLB > 10 with a polynuclear iron (III) polysaccharide complex.

Although examples 1-4 show the positive effect of surfactants with an HLB > 10, the data generated according to Table 10 show an even further improved improvement in the relevant pharmacokinetic parameters.

Example 6:

examples 1-4 have shown the positive effect alone of the addition of surfactants with HLB > 10. Example 5 has shown the positive combined effect of the addition of the surfactant with the polynuclear iron (III) polysaccharide complex in oral applications.

To demonstrate that the overall combined effect is not affected by a particular route of administration, the formulations outlined in table 11 were prepared for administration by injection. The "ref 1" item is the same as the "ref" item outlined in example 5 above, meaning an oral formulation comprising toltrazuril only. The item "ref 2" was the same as test substance 1 of this example, but no active iron was added.

The formulation was prepared similarly to example 5.

TABLE 11 formulation of toltrazuril in Water (with and without iron)

。

In a parallel design, 3 study groups were established, with 6 piglets per group. On the third day of life of the animals, they were treated with each item. The piglets of the group receiving the ref1 formulation received a single oral bolus (exactly as in example 5 above), while piglets of the other two groups each received a single injection of both formulations on day 3 of their life.

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

PK data for table 12 were calculated from the samples obtained.

TABLE 12 pharmacokinetic data for the formulation according to example 6

。

The above data clearly show that the addition of a surfactant with an HLB of ≥ 10 in positive combination with the polynuclear iron (III) polysaccharide complex, irrespective of the route of administration, whereas even higher HLB surfactants appear to further increase the positive effect on the pharmacokinetic profile achieved compared to example 5.

Summarizing the above examples 1-6, it can be determined that surfactants with HLB values of ≥ 10 significantly increase the modified pharmacokinetic profile of the toltrazuril polynuclear iron (III) polysaccharide complex combination, and that such effects are not limited to formulation as a solution or dispersion of the active substance, and that such effects are also influenced by its particular route of administration.