Acid containing lipid formulations

Abstract

The present invention relates to compositions forming a low viscosity mixture of: i) a non-polymeric slow-release matrix ii) at least one biocompatible, (preferably oxygen containing) organic solvent; iii) at least one peptide active agent; and iv) at least one lipid soluble acid. The invention further relates to methods of treatment comprising administration of such compositions, especially in treating diabetes, and to pre-filled administration devices and kits containing the formulations.

Claims

1. A non-aqueous pre-formulation comprising a low viscosity mixture of: a) 30-70 wt. % of at least one neutral diacyl lipid and/or tocopherol; b) 30-60 wt. % of at least one phospholipid; c) at least one biocompatible, organic solvent comprising ethanol; d) at least one peptide active agent other than a GLP-1 receptor agonist; and e) 0.1-5% wt. % of at least one lipid soluble acid selected from methane sulfonic acid or hydrochloric acid; wherein the molar ratio of said peptide active agent to said lipid soluble acid is 1:1 to 1:30; wherein the pre-formulation forms, or is capable of forming, at least one liquid crystalline phase structure upon contact with an aqueous fluid.

2. A non-aqueous pre-formulation as claimed in claim 1 wherein component a) comprises at least one diacyl glycerol; and component b) comprises at least one phosphatidyl choline.

3. A non-aqueous pre-formulation as claimed in claim 1 wherein the organic solvent comprising ethanol is present at a level of 0.1 to 20% by weight.

4. A non-aqueous pre-formulation as claimed in claim 1 wherein the organic solvent comprises ethanol and optionally propylene glycol.

5. A non-aqueous pre-formulation as claimed in claim 1 wherein component c) comprises a mixture of ethanol and an amide, or a mixture of ethanol and a sulfoxide.

6. A non-aqueous pre-formulation as claimed in claim 1 wherein component c) comprises a mixture of ethanol and N-methyl pyrrolidone, or a mixture of ethanol and dimethylsulfoxide.

7. A non-aqueous pre-formulation as claimed in claim 1 which is capable of being dispensed through a needle of 19 awg by manual pressure.

8. A non-aqueous pre-formulation as claimed in claim 1 having a viscosity of 1 to 1000 mPas at 20° C.

9. A method of delivery of a peptide active agent to a human or non-human animal body, said method comprising parenterally administering a non-aqueous pre-formulation of claim 1.

10. A method for the preparation of a depot composition comprising exposing a non-aqueous pre-formulation of claim 1 to an aqueous fluid in vivo.

11. A process for the formation of a non-aqueous pre-formulation according to claim 1 suitable for the administration of a peptide bioactive agent to a subject, said process comprising forming a low viscosity mixture of: i) at least one neutral diacyl lipid and/or tocopherol; ii) at least one phospholipid; iii) at least one biocompatible, organic solvent comprising ethanol; and dissolving or dispersing at least one peptide active agent; and at least one lipid soluble acid in the low viscosity mixture, or in at least one of components i) to iii) prior to forming the low viscosity mixture.

12. A pre-filled administration device containing a pre-formulation as claimed in claim 1.

13. A kit comprising an administration device as claimed in claim 12.

14. A non-aqueous pre-formulation comprising a low viscosity mixture of: a) 30-70 wt. % of at least one neutral diacyl lipid and/or tocopherol; b) 30-60 wt. % of at least one phospholipid c) at least one biocompatible, organic solvent comprising ethanol; d) at least one GLP-1 receptor agonist; and e) 0.1-5 wt. % of methane sulfonic acid; wherein the molar ratio of said peptide active agent to methane sulfonic acid is 1:1 to 1:30; wherein the pre-formulation forms, or is capable of forming, at least one liquid crystalline phase structure upon contact with an aqueous fluid.

15. A non-aqueous pre-formulation as claimed in claim 14 wherein component a) comprises at least one diacyl glycerol; and component b) comprises at least one phosphatidyl choline.

16. A non-aqueous pre-formulation as claimed in claim 14 wherein the organic solvent comprising ethanol is present at a level of 0.1 to 20% by weight.

17. A non-aqueous pre-formulation as claimed in claim 14 wherein the organic solvent comprises ethanol and optionally propylene glycol.

18. A non-aqueous pre-formulation as claimed in claim 14 wherein component c) comprises a mixture of ethanol and an amide, or a mixture of ethanol and a sulfoxide.

19. A non-aqueous pre-formulation as claimed in claim 14 wherein component c) comprises a mixture of ethanol and N-methyl pyrrolidone, or a mixture of ethanol and dimethylsulfoxide.

20. A non-aqueous pre-formulation as claimed in claim 14 which is capable of being dispensed through a needle of 19 awg by manual pressure.

21. A non-aqueous pre-formulation as claimed in claim 14 having a viscosity of 1 to 1000 mPas at 20° C.

22. A method of delivery of a peptide active agent to a human or non-human animal body, said method comprising parenterally administering a non-aqueous pre-formulation of claim 14.

23. A method for the preparation of a depot composition comprising exposing a non-aqueous pre-formulation of claim 14 to an aqueous fluid in vivo.

24. A pre-filled administration device containing a pre-formulation as claimed in claim 14.

25. A kit comprising an administration device as claimed in claim 24.

Description

(1) The Invention will now be further illustrated by reference to the following non-limiting Examples and the attached Figures, in which;

(2) FIG. 1 shows the viscosity reducing effect on addition of solvents.

(3) FIG. 2 displays stability data supporting the highly favourable storage stability obtained with formulation compositions of the invention.

(4) FIG. 3 shows the level of breakdown products in a composition of the invention stored for four weeks at 5° C.

EXAMPLES

Example 1: Availability of Various Liquid Crystalline Phases in the Depot by Choice of Composition

(5) Injectable formulations containing different proportions of phosphatidyl choline (PC) (“SPC”—Lipoid S100) and glycerol dioleate (GDO) and with ethanol (EtOH) as solvent were prepared to illustrate that various liquid crystalline phases can be accessed after equilibrating the depot precursor formulation with excess water.

(6) Appropriate amounts of PC, GDO and EtOH were weighed in glass vials and the mixture was placed on a shaker until the PC completely dissolved to form a clear liquid solution. GDO was then added to form an injectable homogenous solution.

(7) Each formulation was injected in a vial and equilibrated with excess water. The phase behaviour was evaluated visually and between crossed polarizes at 25° C. Results are presented in Table 1.

(8) TABLE-US-00002 TABLE 1 Formulation PC (wt %) GDO (wt %) EtOH (wt %) Phase in H.sub.2O A 22.5 67.5 10.0 L.sub.2 B 28.8 61.2 10.0 I.sub.2 C 45.0 45.0 10.0 I.sub.2/H.sub.II D 63.0 27.0 10.0 H.sub.II/L.sub.α L.sub.2 = reversed micellar phase I.sub.2 = reversed cubic liquid crystalline phase H.sub.II = reversed hexagonal liquid crystalline phase L.sub.α = lamellar phase

Example 2

Viscosity in PC/GDO (5:5) or PC/GDO (4:6) on Addition of Solvent (EtOH, PG and NMP)

(9) A mixture of PC/GDO/EtOH with approximately 25% EtOH was manufactured according to the method in Example 1. All, or nearly all, of the EtOH was removed from the mixture with a rotary evaporator (vacuum, 40° C. for 1 h followed by 50° C. for 2 h) and the resulting mixture was weighed in a glass vial after which 1, 3, 5, 10 or 20% of a solvent (EtOH, propylene glycol (PG) or n-methylpyrrolidone (NMP)) was added. The samples were allowed to equilibrate several days before the viscosity was measured with a CarriMed CSL 100 rheometer equipped with automatic gap setting.

(10) This example clearly illustrates the need for solvent with certain depot precursors in order to obtain an injectable formulation (see FIG. 1). The viscosity of solvent-free PC/GDO mixtures increases with increasing ratio of PC. Systems with low PC/GDO ratio (more GDO) are injectable with a lower concentration of solvent.

Example 3: Degradation of Depot Formulation in the Rat

(11) Various volumes (1, 2, 6 ml/kg) of the depot precursor (36% wt PC, 54% wt GDO, and 10% wt EtOH) were injected in the rat and were removed again after a period of 14 days. It was found that substantial amounts of the formulations were still present subcutaneously in the rat after this time, see Table 3.

(12) TABLE-US-00003 TABLE 3 Dose (ml/kg) Mean diameter day 3 (mm) Mean diameter day 14 (mm) 1 (n = 3) 15.8 12.5 2 (n = 3) 18.5 15.3 6 (n = 3) 23.3 19.3

Example 4: Preparation of a GLP-1 Formulation without pH-Adjusting Agent

(13) The GLP-1 substance and the excipients used in example 4 to 8 are presented in the Table below.

(14) GLP-1 Substance and Excipients Used in the Examples 4 to 8:

(15) TABLE-US-00004 Name Abbreviation Supplier GLP-1(7-36)amide, acetate GLP-1(Ac) PolyPeptide Laboratories, salt Inc., CA, USA Phosphatidylcholine, soy SPC Lipoid, Germany Glycerol dioleate GDO Danisco, Denmark Ethanol (99.5%) EtOH Kemetyl, Sweden Propylene glycol PG Apoteket, Sweden

(16) A lipid formulation comprising 1.08 g SPC, 1.08 g GDO, 0.08 g EtOH and 0.26 g PG was mixed in a 5 mL glass vial (composition: SPC/GDO/EtOH/PG=43.2/43.2/3.2/10.4 wt %). The vial was placed on a mixing table (end-over-end mixing) for approximately 2 hours at RT. A transparent and homogenous formulation was obtained.

(17) 0.02 g of GLP-1(Ac) was weighed into a 2 mL glass vial and 1.98 g of the lipid formulation, prepared as above, was added (giving a total GLP-1(Ac) load of 1 wt %). The formulation was mixed on a vortex mixer (to disperse the GLP-1(Ac) powder in the formulation) and then placed on a mixing table at room temperature for constant end-over-end mixing. After 5 days the sample still contained a lot of undissolved GLP-1(Ac) as assessed visually and the sample was therefore centrifuged at 5000 rpm for 15 minutes to obtain a clear supernatant.

(18) The GLP-1 concentration in the supernatant was assayed by a normal phase (NP) HPLC method using UV detection.

(19) The assayed GLP-1 (equivalents base—GLP-1(0)) concentration in the sample was 6.25 mg/g (0.625 wt %). Because of the long equilibration time used (5 days) this value is taken as the maximum GLP-1 concentration achievable in the lipid formulation without the addition of pH-adjusting agent.

Example 5: Preparation of GLP-1 Formulations with Methane Sulfonic Acid (MeSulf) as pH-Adjusting Agent

(20) A lipid formulation comprising SPC, GDO, EtOH, PG and MeSulf (Sigma-Aldrich, Sweden) was prepared as described in Example 4. The lipid composition was the following: SPC/GDO/EtOH/PG/MeSulf=43.2/43.2/3.0/10.0/0.5 wt %.

(21) The required amount of GLP-1(Ac) powder was weighed into 2 mL glass vials followed by addition of the lipid formulation in an amount appropriate for achieving nominal drug loads of approximately 3 to 6 wt % GLP-1(0). The samples were briefly vortexed followed by continuous end-over-end rotation at room temperature until completely homogenous and transparent samples were obtained (1-3 days).

(22) The concentration of GLP-1 (expressed as equivalents GLP-1 base=GLP-1(0)) in the respective formulations as determined by HPLC is given in the Table below.

(23) GLP-1 Drug Load in Lipid Formulations Containing MeSulf

(24) TABLE-US-00005 Assayed Excess GLP-1 Nominal GLP-1(0) (HPLC) GLP- load compared Formulation conc./wt % 1(0) conc./wt % with Example 4* A 3.00 3.07 4.91 B 3.73 3.77 6.03 C 4.61 4.58 7.33 D 5.43 5.36 8.54 *Calculated as the ratio between the assayed GLP-1(0) concentration with MeSulf as pH-adjusting agent and the concentration found in the formulation without MeSulf (Example 4)

Example 6: Preparation of GLP-1 Formulation with Anhydrous Hydrogen Chloride (HCl) as pH-Adjusting Agent

(25) A lipid formulation comprising SPC, GDO, PG and EtOH.HCl (1.25M HCl in EtOH from Fluka, Sweden) was prepared as described in Example 4. The lipid composition was the following: SPC/GDO/PG/EtOH.HCl=43.0/43.0/10.0/4.0 wt %.

(26) The required amount of GLP-1(Ac) powder was weighed into a 2 mL glass vial followed by addition of the lipid formulation. The sample was briefly vortexed followed by continuous end-over-end rotation at room temperature until a completely homogenous and transparent sample was obtained (1 day).

(27) The concentration of GLP-1 (expressed as GLP-1 base=GLP-1(0)) in the formulation as determined by HPLC is given in the Table below.

(28) GLP-1 Drug Load in Lipid Formulations Containing HCl

(29) TABLE-US-00006 Assayed Excess GLP-1 Nominal GLP-1(0) (HPLC) GLP- load compared Formulation conc./wt % 1(0) conc./wt % with Example 4* E 3.68 3.66 5.86 *Calculated as the ratio between the assayed GLP-1(0) concentration with HCl as pH-adjusting agent and the concentration found in the formulation without HCl (Example 4)

Example 7: Preparation of GLP-1 Formulations Containing Polysorbate 80 (P80) and with Methane Sulfonic Acid (MeSulf) as pH-Adjusting Agent

(30) A lipid formulation comprising SPC, GDO, P80 (Croda, USA), EtOH, PG and MeSulf was prepared as described in Example 4. The lipid composition was the following: SPC/GDO/P80/EtOH/PG/MeSulf=41.0/41.0/5.0/3.0/10.0/0.5 wt %.

(31) The required amount of GLP-1(Ac) powder was weighed into 2 mL glass vials followed by addition of the lipid formulation in an amount appropriate for achieving nominal drug loads of 3, 4, 5 and 6 wt % GLP-1(0). The samples were briefly vortexed followed by continuous end-over-end rotation at room temperature until completely homogenous and transparent samples were obtained (1-3 days) indicating complete dissolution of GLP-1 in the lipid formulation.

Example 8: Stability of GLP-1 in Formulations Containing MeSulf as pH-Adjusting Agent

(32) Lipid formulations containing MeSulf were prepared as described in Example 4.

(33) The required amount of GLP-1(Ac) powder was weighed into 6 mL glass vials followed by addition of the lipid formulation in an amount appropriate for achieving nominal drug loads of 3 wt % GLP-1(0). The samples were briefly vortexed followed by continuous end-over-end rotation at RT until completely homogenous and transparent samples were obtained (1-3 days). The nominal composition of the samples is given in the Table below.

(34) Nominal Composition (Wt %) of Formulations for Stability Study

(35) TABLE-US-00007 Formu- lation GLP-1(0)* SPC DOPC** GDO PG EtOH MeSulf F 3.0 41.7 — 41.7 10.0 3.0 0.5 G 3.0 — 41.7 41.7 10.0 3.0 0.5 *equivalents GLP-1 free base (GLP-1(0)). **Synthetic Dioleoyl Phosphatidylcholine (DOPC) from Lipoid, Germany.

(36) The samples were filled in 1 mL glass vials, capped with Teflon-coated rubber stoppers and stored at 5° C. in a Termak climate chamber. After 4 weeks, the samples were taken out for analysis of GLP-1 content, ID and degradation products using a normal phase HPLC assay and UV detection (214 nm). The results displayed in FIGS. 2 and 3 reveal highly favourable storage stability of the formulations with essentially no degradation (within the error limits of the assay) of GLP-1 during the investigated time period.