INJECTABLE COMPOSITION COMPRISING GnRH ANALOGUE

Abstract

An injectable composition of the present invention has remarkably increased safety by forming, into a composition, a sorbitan unsaturated fatty acid ester comprising g a polar head group with two or more —OH (hydroxyl) groups, is in a liquid phase in a phase where an aqueous fluid is absent so as to be easily applied to pharmaceutical preparations in dosage forms, and forms a liquid crystal in an aqueous fluid phase so that sustained-release effects of GnRH analogue, which is used as a pharmaceutically active substance in vivo, are exhibited. In addition, in an injectable composition of the present invention, a sustained-release lipid initial preparation comprising GnRH analogue includes water, and thus a liquid injectable agent forms a liquid crystal gel immediately after administration, so that the effects of reducing initial release rate and remarkably increasing sustained-release properties of the drug are exhibited.

Claims

1. An injectable composition comprising: a) a sorbitan unsaturated fatty acid ester comprising two or more —OH (hydroxyl) groups in a polar head group; b) a phospholipid; c) a liquid crystal hardener having no ionized group, and a hydrophobic portion thereof having a triacyl group comprising 15 to 40 carbon atoms or a carbon ring structure; d) water; and e) a gonadotropin-releasing hormone (GnRH) analogue as pharmaceutically active sub stance, and wherein the injectable composition is liquid before injection and forms a liquid crystal after injection.

2. The injectable composition according to claim 1, wherein the sorbitan unsaturated fatty acid ester is selected from the group consisting of sorbitan monooleate, sorbitan monolinoleate, sorbitan monopalmitoleate, sorbitan monomyristoleate, sorbitan sesquioleate, sorbitan sesquilinoleate, sorbitan sesquipalmitoleate, sorbitan sesquimyristoleate, sorbitan dioleate, sorbitan dilinoleate, sorbitan dipalmitoleate, sorbitan dimyristoleate, and mixtures thereof.

3. The injectable composition according to claim 1, wherein the sorbitan unsaturated fatty acid ester is selected from the group consisting of sorbitan monooleate, sorbitan sesquioleate, sorbitan monolinoleate, sorbitan monopalmitoleate, sorbitan monomyristoleate, sorbitan sesquioleate, and mixtures thereof.

4. The injectable composition according to claim 1, wherein the phospholipid is selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerine, phosphatidylinositol, phosphatidic acid, sphingomyelin, and mixtures thereof, which have 4 to 30 saturated or unsaturated carbon atoms.

5. (canceled)

6. The injectable composition according to claim 1, wherein the liquid crystal hardener is selected from the group consisting of triglyceride, retinyl palmitate, tocopherol acetate, cholesterol, benzyl benzoate, ubiquinone, and mixtures thereof.

7. (canceled)

8. The injectable composition according to claim 1, wherein the water is added as at least one selected from the group consisting of water for injection, distilled water, and buffer.

9. The injectable composition according to claim 1, wherein the GnRH analogue is a GnRH agonist or a GnRH antagonist, wherein the GnRH agonist is selected from the group consisting of leuprolide, goserelin, triptorelin, nafarelin, buserelin, histrelin, deslorelin, meterelin, gonadrelin, pharmaceutical acceptable salts thereof, and mixtures thereof, and wherein the GnRH antagonist is selected from the group consisting of degarelix, abarelix, ganirelix, cetrorelix, pharmaceutical acceptable salts thereof, and mixtures thereof.

10-15. (canceled)

16. The injectable composition according to claim 1, wherein a weight ratio of component a) and component b) is 10:1 to 1:10.

17. The injectable composition according to claim 1, wherein a weight ratio of components a)+b); and component c) is 1000:1 to 1:1.

18. The injectable composition according to claim 1, wherein a weight ratio of components a)+b)+c); and component d) is 99:1 to 1:1.

19. The injectable composition according to claim 1, wherein a weight ratio of components a)+b)+c)+d); and component e) is 10000:1 to 1:1.

20. The injectable composition according to claim 1, comprising: a) 9 to 90 wt % of the sorbitan unsaturated fatty acid ester having two or more —OH (hydroxyl) groups in a polar head group; b) 9 to 90 wt % of the phospholipid; c) 0.1 to 50 wt % of the liquid crystal hardener which does not have an ionized group, and of which a hydrophobic portion has the triacyl group having 15 to 40 carbon atoms or a carbon ring structure; d) 0.5 to 50 wt % of the water; and e) 0.01 to 50 wt % of gonadotropin-releasing hormone (GnRH) analogue.

21. The injectable composition according to claim 1, comprising: a) 9 to 50 wt % of the sorbitan unsaturated fatty acid ester comprising two or more —OH (hydroxyl) groups in a polar head group; b) 18 to 60 wt % of the phospholipid; c) 1 to 36 wt % of the liquid crystal hardener which does not have an ionized group, and of which a hydrophobic portion has the triacyl group comprising 15 to 40 carbon atoms or a carbon ring structure; d) 0.5 to 10.5 wt % or 25 to 37.5 wt % of the water; and e) 0.1 to 45 wt % of leuprolide or pharmaceutically acceptable salts thereof.

22. The injectable composition according to claim 1, comprising: a) 9 to 50 wt % of the sorbitan unsaturated fatty acid ester comprising two or more —OH (hydroxyl) groups in a polar head group; b) 18 to 60 wt % of the phospholipid; c) 1 to 36 wt % of the liquid crystal hardener which does not have an ionized group, and of which a hydrophobic portion has the triacyl group comprising 15 to 40 carbon atoms or a carbon ring structure; d) 0.5 to 10.5 wt % or 25 to 37.5 wt % of the water; and e) 0.1 to 45 wt % of goserelin or pharmaceutically acceptable salts thereof.

23. The injectable composition according to claim 1, comprising: a) 9 to 50 wt % of the sorbitan unsaturated fatty acid ester comprising two or more —OH (hydroxyl) groups in a polar head group; b) 18 to 60 wt % of the phospholipid; c) 1 to 36 wt % of the liquid crystal hardener which does not have an ionized group, and of which a hydrophobic portion has the triacyl group comprising 15 to 40 carbon atoms or a carbon ring structure; d) 0.5 to 10.5 wt % or 25 to 37.5 wt % of the water; and e) 0.1 to 45 wt % of degarelix or pharmaceutically acceptable salts thereof.

24. A method for preventing or treating sex hormone-dependent diseases, or for contraception, the method, comprising administering a therapeutically effective amount of the injectable composition according to claim 1 into a subject.

25. The method according to claim 24, wherein the sex hormone-dependent disease is prostate cancer, breast cancer, ovarian cancer, endometriosis, uterine fibroid, polycystic ovarian disease, precocious puberty, hypertrichosis, gonadotroph pituitary adenomas, sleep apnea syndrome, irritable bowel syndrome, premenstrual syndrome, benign prostatic hyperplasia, or infertility.

26. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0081] FIG. 1 is a view of pictures showing test results of identifying appearances of compositions according to Examples 1 to 18.

[0082] FIG. 2 is a view of pictures confirming a sol-gel conversion over time in the subcutaneous fat (ex-vivo) of pigs injected with the compositions of Comparative Examples 1 and Example 4, respectively.

[0083] FIG. 3 is a view of pictures identifying appearances of a gel in the mini pigs subcutaneously (in vivo) injected with the compositions of Comparative Example 2 and Example 10, respectively.

[0084] FIG. 4 is a graph showing a PK change up to 7 days in the guinea pigs each injected with the compositions of Comparative Example 3 and Example 13.

[0085] FIG. 5 is a graph showing a PK change up to 28 days in the guinea pigs each injected with the compositions of Comparative Example 3 and Example 13.

[0086] FIG. 6 is a graph showing a PK change of an initial drug release rate in the guinea pigs injected with the composition of Example 19.

[0087] FIG. 7 is a graph showing a PK change up to 28 days in the guinea pigs injected with the composition of Example 19.

[0088] FIG. 8 is a graph showing a PK change up to 7 days in humans each injected with the compositions of Comparative Example 3 and Example 13.

[0089] FIG. 9 is a graph showing a PK change up to 28 days in humans each injected with the compositions of Comparative Example 3 and Example 13.

MODE FOR INVENTION

[0090] Hereinafter, the present invention will be described in more detail through the following examples and experimental examples. However, the following examples and experimental examples are provided only for the purpose of illustrating the present invention, and thus the scope of the present invention is not limited thereto.

[0091] The additives used in the present invention were the excipients according to the standards of the pharmacopoeia and a reagent purchased from Aldrich, Lipoid, Croda, and Seppic (company names).

[Examples 1 to 18] Preparation of Injectable Composition of the Present Invention

[0092] Sorbitan unsaturated fatty acid ester, phospholipid, liquid crystal hardener, water, and pharmacologically active substances were added at the same weight as in Table 1 below.

[0093] Examples 1 to 18 were prepared as follows. First, leuprolide acetate, sorbitan monooleate, phosphatidylcholine, tocopherol acetate, DMSO and ethanol were mixed with a homogenizer (PowerGenmodel125. Fisher) at room temperature for 0.5 hours under the conditions of 1,000 to 3,000 rpm and homogenized to prepare a lipid solution. Then, a corresponding amount of water was added to the prepared lipid solution, and homogenized for about 5 to 30 minutes with a homogenizer under the conditions of about 1,000 to 3,000 rpm, so as to prepare an injectable composition in solution.

TABLE-US-00001 TABLE 1 Example Unit (mg) 1 2 3 4 5 6 7 8 9 Leuprolide acetate 3.75 3.75 3.75 3.75 3.75 3.75 3.75 3.75 3.75 Sorbitan monooleate 22.9 22.2 21.5 20.8 20.1 19.4 18.7 18.1 17.4 Phosphatidylcholine 45.7 44.3 43.0 41.6 40.2 38.8 37.5 36.1 34.8 Tocopherol acetate 15.2 14.8 14.3 13.9 13.4 13.0 12.5 12.0 11.6 DMSO 5 5 5 5 5 5 5 5 5 Ethanol 5 5 5 5 5 5 5 5 5 water 2.5 5 7.5 10 12.5 15 17.5 20.0 22.5 Example Unit (mg) 10 11 12 13 14 15 16 17 18 Leuprolide acetate 3.75 3.75 3.75 3.75 3.75 3.75 3.75 3.75 3.75 Sorbitan monooleate 16.7 16.0 15.3 14.7 13.9 13.3 12.6 11.9 11.2 Phosphatidylcholine 33.4 32.0 30.7 29.3 28.0 26.6 25.2 23.6 22.5 Tocopherol acetate 11.1 10.7 10.2 9.8 9.3 8.9 8.4 7.9 7.5 DMSO 5 5 5 5 5 5 5 5 5 Ethanol 5 5 5 5 5 5 5 5 5 water 25.0 27.5 30.0 32.5 35.0 37.5 40.0 42.5 45

[Example 19] Preparation of Injectable Composition of the Present Invention

[0094] In Example 19, 292 mg of leuprolide acetate, 430 mg of DMSO, 1,034 mg of sorbitan monooleate, 2,195 mg of phosphatidylcholine, 697 mg of tocopherol acetate, and 452 mg of anhydrous alcohol were added to a glass vial and stirred at room temperature to prepare a transparent lipid solution. The prepared lipid solution was filtered through a PVDF Capsule Filter 0.2 μm, and then the weight was measured. Water was injected into the measured lipid solution to obtain a transparent solution, in which a water content might account for 10.5% (w/w) based on 100% by weight of the transparent solution obtained by injecting water, then additionally stirred at room temperature to confirm the transparent solution, and then finally filtered through a PVDF Capsule Filter 0.2 mm so as to prepare an injectable liquid injectable formulation.

Comparative Examples 1 to 3

[0095] Sorbitan unsaturated fatty acid ester, phospholipid, liquid crystal hardener, and pharmacologically active substance were added at the same weight as in Table 2 below.

[0096] In Comparative Examples 1 to 3, leuprolide acetate, sorbitan monooleate, phosphatidylcholine, tocopherol acetate, DMSO and ethanol were mixed with a homogenizer (PowerGenmodel125. Fisher) at room temperature for 0.5 hours under the conditions of 1,000 to 3,000 rpm and homogenized.

TABLE-US-00002 TABLE 2 Comparative Example Unit (mg) 1 2 3 Leuprolide 3.75 3.75 3.75 acetate Sorbitan 22.2 20.8 15.3 monooleate Phosphatidylcholine 44.3 41.6 30.7 Tocopherol 14.8 13.9 10.2 acetate DMSO 5 5 5 Ethanol 5 5 5 water — — —

[Experimental Example 1] Appearance Identification Test

[0097] The appearances of the formulations prepared through Examples 1 to 18 of the present invention were identification. It can be confirmed that the Examples 1 to 4 have transparent appearances when a water content is in the range of 0.5 to 10 wt % based on 100 wt % of the total composition. In addition, it can be confirmed that the appearances of Examples 10 to 15 having a water content of 25 to 37.5 wt % are transparent. This is a section in which water having different physical properties from lipids is stably dissolved without being separated or suspended in the lipid solution.

[Experimental Example 2] Sol-Gel Conversion Test in Pig Subcutaneous Fat (Ex-Vivo)

[0098] A sol-gel conversion test was performed in porcine subcutaneous fatty tissues with the compositions according to Comparative Example 1 and Example 4 of the present invention as follows.

[0099] After slowly injecting 100 uL of the compositions according to Comparative Example 1 and Example 4 into porcine subcutaneous fat, the injection site was cut at 1, 6, 24, 72 and 168 hours after injection to observe a cross-section thereof. The results thereof are shown in [FIG. 2].

[0100] Referring to FIG. 2, in the case of Comparative Example 1 in which water was not added in the porcine subcutaneous fatty tissues, it could be confirmed that the liquid formulation sucks moisture from the tissues and is very slowly converted into a gel. In the case of Example 4 containing water, it could be confirmed that the liquid formulation already contains water, and thus is rapidly converted into a gel immediately after administration.

[0101] As in Comparative Example 1, a drug is rapidly released in a sol state in the body. In the examples according to the present invention, the drug passes through numerous lipid lattice structures and thus is released very slowly after being converted into a gel having a liquid crystal structure.

[Experimental Example 3] Appearance Identification Test of Gel in Mini Pig Under the Skin (In Vivo)

[0102] For Comparative Example 2 and Example 10 of the present invention, the gel appearances in the subcutaneous tissues of a mini pig were identification. The experiment was performed by administering 0.3 mL each of the compositions of Comparative Example 2 and Example 10 by using a disposable syringe under the back skin of a mini pig (male) weighing about 15 kg. In seven days after the administration, an administration site was excised, and the results of comparative observation of the gel appearances are shown in FIG. 3.

[0103] Referring to FIG. 3, the composition according to Comparative Example 2 is a lipid liquid formulation without addition of water, and when converted from sol to gel, the composition is converted to gel at a very slow rate while slowly drawing in body fluid in the tissue. Because the composition is converted to gel very slowly, the sol that has not become a gel permeates between the subcutaneous fats and forms a gel in a fragmented state. It can be confirmed that a gel surface area is widened when the gel is not formed through agglomeration, but is formed in a fragmented state. This may be the cause of a high initial release rate of the drug.

[0104] On the other hand, since the composition of Example 10 according to the present invention is a form already containing water, it can be confirmed that the composition is rapidly converted into a gel in the tissue and agglomerated so that a gel is not formed. It can be seen that the composition of Example 10 according to the present invention may provide a low initial release rate of the drug.

[Experimental Example 4] Confirmation of PK of Injectable Composition in Guinea Pig

[Experimental Example 4-1] Confirmation of PK of Injectable Composition of Example 13, and Comparative Example 3 in Guinea Pig

[0105] A pharmacokinetics of the compositions of the present invention was confirmed in guinea pigs through the following experiment. Using a disposable syringe, the compositions of Comparative Examples 3 and Example 13 were subcutaneously injected into the back of six-week-old male guinea pigs (n=4) which weighed 500 g on average so that an administration weight of leuprolide may reach 3.75 mg/head (equivalent to one month as a human dose). A guinea pig is an animal with developed subcutaneous fat like humans, and an attempt was made to confirm a PK profile (pharmacokinetic profile) in adipose tissues.

[0106] The concentration of leuprolide in a plasma sample of guinea pigs was analyzed with regard to a PK profile for 28 days by using the liquid chromatography/mass spectrometer (LC-MS). The results of [FIG. 4] and [FIG. 5] show an average value for four guinea pigs used in the experiment. [FIG. 4] is to confirm a difference in the initial release rate of the drug, while [FIG. 5] shows a log transformation result in order to confirm a difference in drug blood concentration in guinea pigs in a later half.

[0107] Referring to FIG. 4, in the case of the composition of Example 13 containing water, it was confirmed that a liquid formulation is rapidly converted into a gel in vivo, thereby significantly lowering an initial release rate. For the composition of Comparative Example 3, it can be confirmed that an area under the curve (AUC) up to three days is 7864 μg hr/mL. However, for the composition of Example 13, it can be also confirmed that the AUC up to three days is 5528 μg hr/mL, which is about 30% reduced compared to Comparative Example 3.

[0108] Referring to FIG. 5, it can be confirmed that the composition of Example 13 shows a decline in an initial drug dumping phenomenon and maintains a drug blood concentration higher than the composition of Comparative Example 3 for one month while the drug remains in the gel formulation.

[Experimental Example 4-2] Confirmation of PK of Injectable Composition of Example 19 in Guinea Pig

[0109] Using the liquid injectable formulation prepared in Example 19 according to the present invention, a pharmacokinetics of Example 19 on guinea pigs was confirmed. Using a disposable syringe, the composition of Example 19 was subcutaneously injected into the back of six-week-old male guinea pigs (n=4) which weighed 500 g on average so that an administration weight of leuprolide may reach 3.75 mg/head (equivalent to one month as a human dose). Here, a guinea pig is an animal with developed subcutaneous fat like humans, and an attempt was made to confirm a PK profile (pharmacokinetic profile) in adipose tissues.

[0110] The concentration of leuprolide in a plasma sample of guinea pigs was analyzed with regard to a PK profile for 28 days by using the liquid chromatography/mass spectrometer (LC-MS). The results of [FIG. 6] and [FIG. 7] show an average value for four guinea pigs used in the experiment. [FIG. 6] is to confirm a difference in the initial release rate of the drug, while [FIG. 7] shows a log transformation result in order to confirm a difference in drug blood concentration in guinea pigs in a later half.

[0111] The results of [FIG. 6] and [FIG. 7] show the guinea pig PK results of Example 19 a liquid formulation containing 10.5% of water, and it was confirmed that an initial dumping of the drug is low compared with the PK result value of Comparative Example 3 of Experimental Example 4-1, and it was confirmed that a drug concentration is maintained higher than that of Comparative Example 3 even in 28 days later.

[0112] Through above Experimental Examples 4-1 and 4-2, in the case of the liquid injectable formulations containing water of Examples 19 and 13, it was considered that an initial concentration of the drug is lower than that of the formulation of Comparative Example 3 which does not contain water, and thus safety is more advantageous. And, it was confirmed that a sustained-release property of the drug is improved because the concentration of the drug is maintained higher than that of the composition of Comparative Example 3 even in 28 days later.

[Experimental Example 5] Confirmation of PK of Injectable Composition in Humans

[0113] For each of the compositions of Comparative Example 3 and Example 13, a pharmacokinetics of Example 13 in human was confirmed. For healthy postmenopausal women, 3.75 mg of Comparative Example 3 and Example 13 as leuprolide was subcutaneously administered to the abdomen of six patients per test group. The concentration of leuprolide in a plasma sample of humans was analyzed with regard to a PK profile for 28 days by using the liquid chromatography/mass spectrometer (LC-MS). The PK profiles of Comparative Example 3 and Example 13 in humans were as shown in FIG. 8 and [FIG. 9]. [FIG. 8] is to confirm a difference in the initial drug release rate, and [FIG. 9] shows a log-transformation to confirm a difference in the drug blood concentration of the leuprolide drug in the second half.

[0114] Referring to FIG. 8, in the PK profile in humans similar to that of the guinea pig, it was confirmed that a drug dumping phenomenon immediately after administration is remarkably lowered in the composition of Example 13 compared to the composition of Comparative Example 3.

[0115] For the composition of Comparative Example 3, it was found that an area under the curve (AUC) up to three days after administration is 276 μg hr/mL. However, for that of Example 13, it was found that the AUC up to three days is 209 μg hr/mL, which is about 25% lower than the composition of Comparative Example 3.

[0116] Referring to FIG. 9, it was found that the composition of Example 13 shows a decline in an initial drug dumping phenomenon and maintains a drug blood concentration higher than the composition of Comparative Example 3 for one month while the drug remains in the gel formulation.