Compositions suitable for administration of creatine esters

Abstract

The present invention relates to a composition in the form of a particulate emulsion comprising: at least one creatine ester or creatine ester salt in particulate form; 50% (w / w) to 80% (w / w) of an aqueous solvent; from 10% (w / w) to 40% (w / w) of a lipid.

Description

Technical Field

[0001] This invention relates to compositions that can be used for the administration of creatine esters or creatine ester salts. Background Technology

[0002] Creatine is an endogenous nutrient obtained from food and is also naturally produced by the liver and kidneys in most vertebrates. Creatine is also known as 2-[formamidinyl(methyl)amino]acetic acid. It is represented by the nominal formula NH2-C(NH)-N(CH3)-CH2-COOH and has the following structure:

[0003]

[0004] Creatine is primarily used as a supplement to increase muscle mass and enhance muscle performance.

[0005] Creatine is primarily found in muscles, but it is also present in the brain, eyes, testes, and kidneys. In particular, creatine is essential for proper brain function, plays a crucial role in energy storage and transport, and also has additional functions as an anti-apoptotic agent, antioxidant, neuroprotective agent, and neuromodulator.

[0006] However, when creatine monohydrate is administered as an oral supplement, brain enrichment is poor, primarily due to creatine transport proteins that restrict transfer from the blood to the brain and exhibit high inter-individual variability. Therefore, neuronal supplementation with creatine remains a challenge.

[0007] Creatine deficiency involves defects in its biosynthesis via enzymes such as arginine:glycine amidotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT), as well as defects in its transcellular transport, particularly at the brain level. The latter, known as creatine transporter deficiency (CTD), is a neurological disorder caused by damage to the cellular creatine transporter (CrT) encoded by SLC6A8. This condition is inherited in 70% of cases. It significantly affects brain function; intellectual disability, severe speech delay, autism-like behavior, and seizures are the main phenotypic outcomes, but other symptoms can also be observed, such as developmental arrest, hypotonia, neurological and psychiatric complications, and cardiac and gastrointestinal manifestations. CTD is a rare disease with approximately 20,000 possible cases, but only a few hundred have been reported in the medical literature to date.

[0008] Various treatments have been experimented with worldwide to address the critical issue of creatine deficiency in the brain cells of patients with CTD. Currently, none of these methods have yielded significant benefits for CTD patients. In fact, oral administration of creatine monohydrate does not produce any improvement because the compound cannot penetrate the brain cells of CTD patients. Administration of creatine precursors has also failed, indicating that this treatment attempt, while effective for other creatine deficiencies (such as AGAT and GAMT), is unsuitable for treating CTD patients.

[0009] Creatine prodrugs such as creatine fatty acid esters have yielded promising results in animals and in vitro using patient cells. Among them, dodecyl creatine ester (DCE) has shown to be a good treatment option for patients with CTD (Trotier-Faurion et al. (2013) J.Med.Chem.56:5173-81; Ullio-Gamboa et al. (2019) Nanomedicine 14:1579-12593).

[0010] However, in biological fluids such as plasma or extracellular fluid, creatine esters are rapidly degraded by esterases and highly efficient non-enzymatic cyclization, as intramolecular nucleophilic reactions result in ineffective creatine anhydride. This cyclization prevents the use of creatine esters (e.g., creatine ethyl ester) as an effective supplement for CTD.

[0011] Therefore, an effective delivery system is needed to overcome the rapid degradation of creatine prodrugs in plasma and extracellular fluids in order to address CTD and other brain energy deficiencies.

[0012] Therefore, in international publication WO2020 / 221780, it has been proposed to formulate creatine esters or their salts in a composition comprising at least one ω-3 fatty acid or its salt and at least one glyceride for use in a pharmaceutical, said composition being free of nonionic surfactants.

[0013] However, the amount of creatine that can be loaded and its stability in the composition are poor, the latter being due to cyclization into ineffective creatine anhydride, which prevents the formulation from being used as a drug. Furthermore, the composition is not particularly suitable for administration by spray, which is the most practical form of intranasal administration of creatine.

[0014] Therefore, formulations containing creatine esters, which are stable and can be loaded in large quantities, are still needed. Summary of the Invention

[0015] This invention stems from an accidental discovery by the inventors that a lipid-based particulate emulsion in water containing dodecyl creatine (DCE) in particulate form exhibits high stability and can be loaded with large amounts of DCE. Furthermore, this particulate emulsion is suitable for spray administration and can efficiently deliver creatine or creatine esters to the brain via the nasal-to-brain route.

[0016] Therefore, the present invention relates to a composition in the form of a particulate emulsion, comprising:

[0017] - At least one creatine ester or creatine ester salt in particulate form;

[0018] -50% (w / w) to 80% (w / w) of aqueous solvents;

[0019] -10% (w / w) to 40% (w / w) of lipids.

[0020] In one embodiment, the present invention relates to a composition in the form of a particulate emulsion, comprising:

[0021] - At least one creatine ester or creatine ester salt in particulate form;

[0022] -50% (w / w) to 80% (w / w) of aqueous solvents;

[0023] -10% (w / w) to 40% (w / w) lipids;

[0024] - At least one surfactant, wherein the concentration of the at least one surfactant is less than 15% (w / w), that is, the total concentration of one or more surfactants contained in the composition is less than 15% (w / w).

[0025] Advantageously, the compositions according to the invention can contain more creatine esters or creatine ester salts than prior art creatine-containing compositions for intranasal administration, i.e., the loading capacity of the compositions according to the invention is increased. Furthermore, the compositions according to the invention allow for increased stability of creatine esters or creatine ester salts. In addition, the compositions of the invention are suitable for spray administration and can deliver more creatine or creatine esters to the brain via the nasal route compared to prior art creatine-containing compositions for intranasal administration.

[0026] The present invention also relates to a method for preparing a composition in the form of a particulate emulsion, the composition comprising at least one particulate creatine ester or creatine ester salt, the method comprising:

[0027] - A solution of at least one surfactant is added to a first mixture of at least one creatine ester or creatine ester salt and lipids to form a second mixture.

[0028] - Stir the second mixture to form a particulate emulsion comprising at least one particulate form of creatine ester or creatine ester salt.

[0029] The concentration of the at least one surfactant in the composition in particulate emulsion form is less than 15% (w / w). Invention Details

[0031] As expected in this document, the word "contains" is synonymous with "including" or "containing". When a subject is stated to include one or more features, it means that features other than those mentioned can be included in the subject. Conversely, the expression "consisting of" is synonymous with "composed of". When a subject is stated to consist of one or more features, it means that features other than those mentioned are not included in the subject.

[0032] Composition

[0033] Emulsions, particularly nanoemulsions or microemulsions, are well known to those skilled in the art and are specifically described in Ullio-Gamboa et al. (2019) Nanomedicine 14:1579-1593, which is incorporated herein by reference.

[0034] Preferably, the composition according to the invention does not contain ω-3 fatty acids, such as DHA.

[0035] Preferably, the composition according to the invention further comprises at least one humectant. More preferably, the humectant is selected from talc, silica, vegetable stearin, magnesium stearate, glycerin, stearic acid, and hyaluronic acid salts (e.g., sodium hyaluronate), particularly glycerin and / or sodium hyaluronate. Most preferably, the composition comprises 1% (w / w) to 10% (w / w), 2% (w / w) to 7% (w / w), 2.5% (w / w) to 5.5% (w / w), or 3% (w / w) to 5% (w / w) of humectant.

[0036] Preferably, the composition according to the invention further comprises at least one stabilizer. As will be apparent to those skilled in the art, the term stabilizer relates to emulsion stabilizers. More preferably, the stabilizer is selected from β-lactoglobulin, gum arabic, and xanthan gum, and particularly, the stabilizer is xanthan gum. Most preferably, the composition comprises 0.05% (w / w) to 1% (w / w), 0.1% (w / w) to 0.5% (w / w), or 0.2% (w / w) to 0.4% (w / w) of the stabilizer.

[0037] Preferably, the composition according to the invention further comprises at least one surfactant, particularly at least one nonionic surfactant. More preferably, the surfactant is selected from polysorbates (e.g., polysorbate 20 or polysorbate 80), cocosides, glycol esters, polyol esters such as dehydrated sorbitol esters (e.g., Span 80), lecithin, cholesterol, polyethylene glycol and its derivatives, polyglycerol and creatine dodecyl ester, and particularly, the surfactant is polysorbate 80. More preferably, the composition according to the invention comprises at least one surfactant (i.e., the total amount of one or more surfactants contained in the composition) at a concentration of less than 15% (w / w), 10% (w / w), 9% (w / w), 8% (w / w), 7% (w / w), 6% (w / w), 5% (w / w), 4% (w / w), 3% (w / w), 2% (w / w), 1% (w / w), 0.5% (w / w), or 0.2% (w / w). Most preferably, the composition comprises 0.01% (w / w) to 1% (w / w), 0.05% (w / w) to 0.2% (w / w), or 0.09% (w / w) to 0.11% (w / w) of a surfactant.

[0038] Preferably, the composition according to the invention further comprises sodium chloride (NaCl). More preferably, the composition comprises 0.1% (w / w) to 2% (w / w), 0.5% (w / w) to 1.5% (w / w), or 0.7% (w / w) to 1.0% (w / w) of NaCl.

[0039] Preferably, the aqueous solvent as defined above is water. More preferably, the water is purified water, distilled water, or deionized water. Most preferably, the composition comprises 55% (w / w) to 75% (w / w), 60% (w / w) to 70% (w / w), or 64% (w / w) to 66% (w / w) of water.

[0040] "Lipids" are well known to those skilled in the art and refer to hydrophobic or amphiphilic small molecules, including in particular fats, waxes, sterols, fat-soluble vitamins, fatty acids, their salts and esters, monoglycerides, diglycerides, triglycerides, and phospholipids.

[0041] Preferably, the lipid as defined above is a triglyceride, particularly a medium-chain triglyceride, and / or isopropyl myristate. More preferably, the lipid is a medium-chain triglyceride. Even more preferably, the lipid is also isopropyl myristate.

[0042] Medium-chain triglycerides are well known to those skilled in the art. Preferably, a medium-chain triglyceride is a triglyceride having two or three fatty acids, said fatty acids being straight-chain or branched, saturated or unsaturated aliphatic chains having 6 to 12 carbon atoms. Preferably, the fatty acids are selected from caprylic acid, octanoic acid, capric acid, and lauric acid.

[0043] Preferably, the composition comprises 15% (w / w) to 35% (w / w), 20% (w / w) to 30% (w / w), or 24% (w / w) to 26% (w / w) of medium-chain triglycerides and / or isopropyl myristate. More preferably, the composition comprises 15% (w / w) to 35% (w / w), 20% (w / w) to 30% (w / w), or 24% (w / w) to 26% (w / w) of medium-chain triglycerides. More preferably, the composition comprises 15% (w / w) to 35% (w / w), 20% (w / w) to 30% (w / w), or 24% (w / w) to 26% (w / w) of isopropyl myristate.

[0044] At least one preservative, such as benzalkonium chloride, may also be included in the pharmaceutical composition as defined above.

[0045] The pharmaceutical compositions according to the invention may further comprise flavoring agents (e.g., imparting a strawberry, banana, vanilla, or orange blossom flavor), flavor masking agents, or flavor enhancers, particularly at concentrations of 0.001% to 0.01%, and more particularly at concentrations of 0.002% to 0.003%.

[0046] Preferably, the composition according to the invention is in the form of a particulate emulsion and comprises:

[0047] Creatine esters or creatine ester salts in particulate form, ranging from -0.1% (w / w) to 10% (w / w);

[0048] -64% (w / w) to 66% (w / w) water;

[0049] Medium-chain triglycerides ranging from -24% (w / w) to 26% (w / w).

[0050] Creatine

[0051] Preferably, the creatine ester as defined above has the following formula (I):

[0052] (NH2)-C(NH)-N(CH3)-CH2-COOR (I)

[0053] Where R represents:

[0054] - Hexosyl, particularly glucosyl, which is optionally substituted by at least one group selected from the following: alkyl containing 1 to 5 carbon atoms, aryl containing 5 to 10 carbon atoms, alkoxy containing 1 to 5 carbon atoms, halogen atom, cyano, trifluoromethyl or nitro.

[0055] - An alkyl group comprising 1 to 30 carbon atoms, wherein the alkyl group is optionally substituted with at least one group selected from the following: an alkyl group comprising 1 to 5 carbon atoms, an aryl group comprising 5 to 10 carbon atoms, an alkoxy group comprising 1 to 5 carbon atoms, a halogen atom, a cyano group, a trifluoromethyl group, or a nitro group.

[0056] - An alkenyl group comprising 2 to 30 carbon atoms, optionally substituted with at least one group selected from: an alkyl group comprising 1 to 5 carbon atoms, an aryl group comprising 5 to 10 carbon atoms, an alkoxy group comprising 1 to 5 carbon atoms, a halogen atom, a cyano group, a trifluoromethyl group, or a nitro group; or

[0057] - An aryl group comprising 5 to 30 carbon atoms, wherein the aryl group is optionally substituted by at least one group selected from the following: an alkyl group comprising 1 to 5 carbon atoms, an aryl group comprising 5 to 10 carbon atoms, an alkoxy group comprising 1 to 5 carbon atoms, a halogen atom, a cyano group, a trifluoromethyl group, or a nitro group.

[0058] Preferably, R represents an alkyl group containing 6 to 20 carbon atoms, more preferably 8 to 15 carbon atoms.

[0059] Preferably, the creatine esters as defined above are selected from nonyl creatine ester (NCE), decyl creatine ester, undecyl creatine ester, dodecyl creatine ester (DCE), tridecyl creatine ester and tetradecyl creatine ester.

[0060] Preferably, the creatine ester as defined above is dodecyl creatine ester (DCE) and / or nonyl creatine ester (NCE). More preferably, the creatine ester as defined above is dodecyl creatine ester (DCE). More preferably, the creatine ester as defined above is nonyl creatine ester (NCE).

[0061] Preferably, the creatine ester salt is selected from salts of alkali metals (especially sodium or potassium), salts of alkaline earth metals (especially magnesium or calcium), salts of organic amides (more particularly amino acids such as arginine or lysine), salts of inorganic acids (such as hydrochloric acid, sulfuric acid or hydrobromic acid), and salts of organic acids (such as acetic acid, formic acid, fumaric acid, trifluoromethanesulfonic acid, tartaric acid, oxalic acid, citric acid, trifluoroacetic acid or methanesulfonic acid).

[0062] As expected in this article, creatine esters or creatine ester salts are in particulate form, that is, creatine esters or creatine ester salts are suspended as particles in the emulsion.

[0063] Preferably, the creatine ester or creatine ester salt as defined above is micronized.

[0064] Preferably, the volume average diameter (Dv) of the creatine or creatine salt particles is about 0.1 μm to about 100 μm.

[0065] Preferably, at least 90% of the creatine ester or creatine salt particles have a volume average diameter (Dv) equal to or greater than 1 μm, 2 μm, 3 μm, 4 μm or 5 μm.

[0066] Preferably, less than 10% of the creatine or creatine salt particles have a volume average diameter (Dv) equal to or less than 1 μm, 2 μm, 3 μm, 4 μm or 5 μm.

[0067] Preferably, at least 90% of the creatine ester or creatine ester salt particles have a volume average diameter (Dv) equal to or less than 50 μm, 30 μm or 25 μm.

[0068] Preferably, less than 10% of the creatine ester or creatine ester salt particles have a volume average diameter (Dv) equal to or greater than 50 μm, 30 μm or 25 μm.

[0069] Preferably, at least 50% of the creatine ester or creatine salt particles have a volume average diameter (Dv) equal to or greater than 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 11 μm, 12 μm, 13 μm, 14 μm or 15 μm.

[0070] Preferably, less than 50% of the creatine ester or creatine salt particles have a volume average diameter (Dv) equal to or less than 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 11 μm, 12 μm, 13 μm, 14 μm or 15 μm.

[0071] Preferably, the volume average diameter (Dv) distribution of the creatine ester or creatine ester salt particles is such that:

[0072] Dv (10) is 2 to 6 μm, preferably 2 to 4 μm, and more preferably 2 to 3 μm;

[0073] Dv (50) is 7 to 13 μm, preferably 7 to 10 μm, more preferably 7 to 9 μm; and

[0074] Dv (90) is 20 to 30 μm.

[0075] Preferably, the average width (W) of the creatine or creatine salt particles is from about 0.1 μm to about 100 μm.

[0076] Preferably, at least 90% of the creatine or creatine salt particles have an average width (W) equal to or greater than 0.5 μm, 2.5 μm or 8 μm.

[0077] Preferably, less than 10% of the creatine or creatine salt particles have an average width (W) equal to or less than 0.5 μm, 2.5 μm or 8 μm.

[0078] Preferably, at least 90% of the creatine ester or creatine ester salt particles have an average width (W) equal to or less than 6 μm, 8 μm or 25 μm.

[0079] Preferably, less than 10% of the creatine or creatine salt particles have an average width (W) equal to or greater than 6 μm, 8 μm or 25 μm.

[0080] Preferably, at least 50% of the creatine or creatine salt particles have an average width (W) equal to or greater than 2.75 μm, 5 μm or 15 μm.

[0081] Preferably, less than 50% of the creatine or creatine salt particles have an average width (W) equal to or less than 2.75 μm, 5 μm or 15 μm.

[0082] Preferably, the average width (W) distribution of creatine ester or creatine ester salt particles is such that:

[0083] W (10) is 0.5 to 8 μm, preferably 2 to 7 μm, and more preferably 0.5 to 2.5 μm;

[0084] W (50) is 2.75 to 15 μm, preferably 6 to 12 μm, more preferably 2.75 to 5 μm; and

[0085] W (90) is 6 to 25 μm, preferably 10 to 22 μm, and more preferably 6 to 8 μm.

[0086] Preferably, the average length (L) of the creatine or creatine salt particles is from about 0.1 μm to about 100 μm.

[0087] Preferably, at least 90% of the creatine or creatine salt particles have an average length (L) equal to or greater than 0.5 μm, 2.5 μm or 7 μm.

[0088] Preferably, less than 10% of the creatine or creatine salt particles have an average length (L) equal to or less than 0.5 μm, 2.5 μm or 7 μm.

[0089] Preferably, at least 90% of the creatine ester or creatine ester salt particles have an average length (L) equal to or less than 18 μm, 22 μm or 40 μm.

[0090] Preferably, less than 10% of the creatine or creatine salt particles have an average length (L) equal to or greater than 18 μm, 22 μm or 40 μm.

[0091] Preferably, at least 50% of the creatine or creatine salt particles have an average length (L) equal to or greater than 5 μm, 9 μm or 20 μm.

[0092] Preferably, less than 50% of the creatine or creatine salt particles have an average length (L) equal to or less than 5 μm, 9 μm or 20 μm.

[0093] Preferably, the average length (L) distribution of creatine ester or creatine ester salt particles is such that:

[0094] L (10) is 0.5 to 7 μm, preferably 0.5 to 2.5 μm;

[0095] L (50) is 5 to 20 μm, preferably 5 to 9 μm; and

[0096] L (90) is 18 to 40 µm, preferably 18 to 22 µm.

[0097] Methods for determining the volume-average diameter (Dv), average width (W), average length (L), and the distribution of Dv, W, and L of particles are well known to those skilled in the art, and notably involve microscopy techniques, particularly by a Malvern Morphologi 4 or Morphology G3S device, or laser diffraction techniques, particularly by a Malvern Mastersizer 3000 Hydro MV device.

[0098] Preferably, the composition according to the invention comprises 0.1% (w / w) to 10% (w / w), 1% (w / w) to 8% (w / w) or 2% (w / w) to 6% (w / w) of creatine or creatine salt.

[0099] Preferably, the composition according to the invention comprises:

[0100] -3 to 5% (w / w) micronized DCE, HCl salt;

[0101] -60 to 70% (w / w) pure water;

[0102] -0.5 to 1.0% (w / w) sodium chloride;

[0103] -1.0 to 6.0% (w / w) glycerol (4811);

[0104] -0.1 to 0.3% (w / w) sodium hyaluronate;

[0105] -20 to 30% (w / w) triglyceride-based lipids (Kollisolv® MCT 70, BASF);

[0106] -0.1 to 0.5% (w / w) xanthan gum;

[0107] -0.05% to 0.20% (w / w) polysorbate 80,

[0108] -Optional 0.01 to 0.05% (w / w) neroli oil.

[0109] Uses in treatment

[0110] Preferably, the composition according to the invention is a pharmaceutical composition or a medical device.

[0111] Preferably, the pharmaceutical composition or medical device as defined above further comprises at least one pharmaceutically acceptable medium, carrier or excipient, particularly a medium, carrier or excipient suitable for application via a local or nasal route.

[0112] Preferably, the composition according to the invention is suitable for spray application. The sprayer may be a single-dose, dual-dose, or multi-dose device.

[0113] Preferably, the composition according to the invention is a dermatological composition.

[0114] Preferably, the compositions according to the invention, particularly dermatological compositions, are suitable for topical application, especially by means of sprays, creams, ointments, gels or balms.

[0115] Preferably, the composition according to the invention is administered in unit doses of 50 μL to 2 mL.

[0116] Preferably, the composition according to the invention is used as a medicine.

[0117] Preferably, the dermatological composition according to the invention is used as a dermatological medicine.

[0118] Preferably, the compositions according to the invention are used for the prevention or treatment of creatine deficiency, such as creatine transporter deficiency (CTD) syndrome, GAMT deficiency or AGAT deficiency, mitochondrial diseases such as Leich syndrome, Leber hereditary optic neuropathy or MELAS syndrome, neurodegenerative diseases, especially Parkinson's disease and related diseases, or multiple system atrophy (MSA), neuromuscular diseases, hypoxia, ischemic encephalopathy such as stroke, cardiovascular diseases, muscular dystrophy, skin diseases such as atopic dermatitis or skin lesions, amyotrophic lateral sclerosis, adrenoleukodystrophy, leukodystrophy, Rett syndrome, Kabuki syndrome or inflammation.

[0119] More preferably, the composition according to the invention is used for the prevention or treatment of creatine transporter deficiency (CTD).

[0120] Uses in cosmetics

[0121] Preferably, the composition according to the invention is a cosmetic composition.

[0122] The present invention also relates to the use of the compositions according to the invention as cosmetics or for cosmetic applications.

[0123] When used as a cosmetic composition, the composition according to the invention is preferably applied topically to the skin of an individual, more preferably a human individual, particularly by means of a spray, cream, ointment, gel, or balm. The cosmetic use of the composition according to the invention aims to improve the appearance of the skin to which it is applied. In this case, the individual preferably does not suffer from any disease, particularly a skin condition, that can be treated with the composition according to the invention.

[0124] individual

[0125] Preferably, the composition according to the invention is applied to an individual, said individual being a mammal, particularly a human.

[0126] method

[0127] Preferably, the solution containing at least one surfactant also contains NaCl and / or at least one wetting agent. The solvent of the solution is an aqueous solvent, preferably water.

[0128] Preferably, a solution of at least one stabilizer is mixed with the particulate emulsion.

[0129] The invention will be further described with reference to the following non-limiting drawings and embodiments. Attached Figure Description

[0130] Figure 1 The measured values ​​of the mist cone (spray) angle (vertical axis) in degrees (RT: room temperature) of four test pumps using the composition in particulate emulsion form according to the invention are shown.

[0131] Figure 2 The ellipticity (vertical axis) of the spray (RT: room temperature) is represented by four test pumps using the composition in the form of a particulate emulsion according to the invention.

[0132] Figure 3 The brain concentration (vertical axis, in ng / g) of DCE in SLC6A8 KO mice in which the intranasal carrier (KO-carrier), the non-particulate DCE formulation (KO-4.4s), or the DCE composition in particulate emulsion form according to the invention (KO-510) is indicated.

[0133] Figure 4 The brain concentration of creatine in SLC6A8 KO mice (vertical axis, in ng / g) is represented by the intranasal administration of the carrier (KO-carrier), the non-particulate DCE formulation (KO-4.4s), or the DCE composition in particulate emulsion form according to the invention (KO-510).

[0134] Figure 5 Microscopic view (upward, polarized) of the composition in particulate emulsion form according to the invention, wherein the DCE concentration is 40 mg / g. Example

[0135] 1. Preparation of particulate emulsion of dodecyl creatine (DCE)

[0136] Prepare a composition having the following components:

[0137] Components % (w / w) Function Micronized DCE, HCl salt 3 to 5 API purified water 60 to 70 Aqueous phase Sodium chloride 0.5 to 1.0 Penetrant Glycerin (4811) Versions 1.0 to 6.0 wetting agent Optional sodium hyaluronate 0.1 to 0.3 wetting agent Triglyceride-based lipids (Kollisolv® MCT70, BASF) 20 to 30 oil phase Xanthan Gum 0.1 to 0.5 stabilizer Polysorbate 80 0.05 to 0.20 surfactants Choose any orange blossom oil 0.01 to 0.05 Flavor masking agent

[0138] Table 1: Compositions according to the invention (for 100g)

[0139] In short, the first mixture (mixture 1) is prepared as follows:

[0140] In a beaker, dissolve NaCl in water. Add glycerol and polysorbate 80, and stir with a magnetic stir bar until completely dissolved to obtain a homogeneous mixture.

[0141] The second mixture (mixture 2) is prepared as follows:

[0142] Dissolve DCE hydrochloride in medium-chain triglycerides in a beaker. Use a magnetic stir bar to stir until completely dissolved to obtain a homogeneous mixture.

[0143] Mixture 1 is mixed with mixture 2 under stirring (magnetic stirring rod) to obtain a third mixture (mixture 3) as follows:

[0144] Place an Ultraturax in a beaker containing the aqueous phase (mixture 1), and gently pour in the oil phase (mixture 2) while stirring with the Ultraturax. Mix until a white emulsion is obtained. Then gradually reduce the speed of the Ultraturax until it stops.

[0145] The fourth mixture (mixture 4) is prepared as follows:

[0146] In a beaker, use a magnetic stir bar to dissolve xanthan gum in water until it is completely dissolved.

[0147] Finally, mix mixture 3 with mixture 4 to obtain a homogeneous emulsion as follows:

[0148] Gently pour mixture 4 into mixture 3 while stirring with an Ultraturax mixer. Add the flavor masking agent. Mix until a complete and homogeneous emulsion is formed. Allow it to stand and perform microscopic monitoring to check the emulsion homogeneity. Figure 5 ).

[0149] Alternatively, the first mixture (mixture 1) can be prepared as follows:

[0150] In a beaker, dissolve NaCl in water. Then, using a magnetic stir bar, dissolve xanthan gum in the aqueous phase until completely dissolved. Add glycerol and polysorbate 80, and use a magnetic stir bar until completely dissolved to obtain a homogeneous mixture.

[0151] The second mixture (mixture 2) is prepared as follows:

[0152] Dissolve DCE hydrochloride in medium-chain triglycerides in a beaker. Use a magnetic stir bar to stir until completely dissolved to obtain a homogeneous mixture.

[0153] Mixture 1 is mixed with mixture 2 under stirring (magnetic stirring rod) to obtain a third mixture (mixture 3) as follows:

[0154] Place an Ultraturax in a beaker containing the aqueous phase (mixture 1), and gently incorporate the oil phase (mixture 2) while stirring with the Ultraturax. Mix until a homogeneous white particulate emulsion is obtained. Then gradually reduce the speed of the Ultraturax until it stops. Add the flavor masking agent. Mix until a complete and homogeneous emulsion is formed. Allow it to stand and examine the uniformity of the particulate emulsion under a microscope. Figure 5 ).

[0155] 2. Spray application of the composition

[0156] The composition is sprayed using several pumps as follows:

[0157] pump Manufacturer Delivery volume CPS APTAR 100 μL 3K URSATEC 100 μL PFP N SILGAN 140 μL VP7 CB18 APTAR 100 μL

[0158] Sprayability is evaluated by calculating the fog cone (spray) angle, fog cone (spray) ellipticity, and delivery uniformity.

[0159] 2.1. Spray Angle

[0160] The spray angle is determined by the tangent of the spray cone at a distance of 60 mm from the nozzle tip.

[0161] The results of mist cone (spray) angle measurements (in degrees) using four pumps of granular emulsion are shown below. Figure 1 And in Table 1 below:

[0162]

[0163] RT = room temperature

[0164] Seeking large spray angles. In this embodiment, the particulate emulsion used for spraying allows for sprays with large angles for all pumps, with the SILGAN PFP N pump exhibiting a large spray angle and the APTAR VP7CB18 pump also effective for spraying particulate emulsions.

[0165] 2.2. Spray ellipticity

[0166] The spray ellipticity is obtained from the spray pattern calculated at a distance of 6 cm from the pump tip. The spray ellipticity is calculated using the following formula: Dmax / Dmin.

[0167] exist Figure 2 The spray ellipticity measurements for four pumps used with the particulate emulsion at different temperatures are shown in Table 2 below:

[0168]

[0169] The goal was to achieve an ellipticity close to 1. In this embodiment, good quality spray ellipticity was obtained with all tested pumps, with the APTAR VP7-CB18 pump showing a slight advantage.

[0170] 2.3. Uniformity of delivery

[0171] - Uniformity of delivery dose with a fully emptied pump

[0172] The delivered dose was weighed throughout the pump's lifespan. Each experiment was conducted by two different investigators to account for potential inter-individual variability. Some experiments were performed at room temperature or after the device had been stored at 4°C for at least 24 hours.

[0173] CPS (APTAR), VP7 CB18 (APTAR), and 3K (URSATEC) deliver a consistent dose during the first 70 actuations.

[0174] - Uniformity of delivery dosage according to the European Pharmacopoeia

[0175] For granular emulsions combined with three pumps (APTAR CPS, APTAR VP7 CB 18, and APTAR CPS), the uniformity of DCE delivery dose was measured. This test was performed according to the European Pharmacopoeia for nasal formulations.

[0176] The DCE content was quantified in the delivered formulation during 10 pump actuations (3 at the beginning of pump life, 4 in the middle, and 3 at the end of pump life). The results are listed in Tables 3-5 below:

[0177]

[0178] Table 3: Uniformity of DCE dose delivered by APTAR CPS pump according to the European Pharmacopoeia (bold values ​​are within acceptable ranges, italic values ​​are outside acceptable ranges)

[0179]

[0180] Table 4: Uniformity of DCE dose delivered by the APTAR VP7 CB18 pump according to the European Pharmacopoeia (bold values ​​are within acceptable ranges, italic values ​​are outside acceptable ranges)

[0181]

[0182] Table 5: Uniformity of DCE dose delivered by APTAR CPS pump according to the European Pharmacopoeia (bold values ​​are within acceptable ranges, italic values ​​are outside acceptable ranges)

[0183] The acceptance criteria for nasal preparations in the European Pharmacopoeia are:

[0184] - The average delivered dose was between 85% and 115% of the target dose;

[0185] - Actuation occurs at most once outside the 75%-125% range of the target, and no actuation occurs outside the 65%-135% range.

[0186] All three combinations of the tested pump / granule emulsion met all the criteria.

[0187] 4. Stability of DCE in particulate emulsions

[0188] The chemical stability of DCE in particulate emulsions was monitored using HPLC / UV methods at 4 °C and room temperature. The results are shown in Tables 6-7 below.

[0189]

[0190] Table 6: Stability of DCE (initial content %) in two batches of particulate emulsion stored at 4°C and estimated values ​​after 18 months of storage.

[0191] DCE is stable when stored in a refrigerator (4°C) for more than 3 months, and extrapolation using the decay slope indicates that more than 95% should be retained after 18 months.

[0192]

[0193] Table 7: Stability of DCE (initial content %) in two batches of particulate emulsion stored at room temperature (20-25℃) and estimated values ​​after 1 month of storage.

[0194] DCE is stable when stored at room temperature (20-25°C) for more than 20 days, and extrapolation using the decay slope shows that 99% remains after 1 month.

[0195] In view of the above, the particulate emulsion according to the present invention is suitable for long-term storage in a refrigerator. Its good short-term stability at room temperature allows the particulate emulsion to be stored outside the refrigerator for at least one month.

[0196] 5. The effect of delivering creatine to the brain

[0197] Three groups of SLC6A8 knockout (KO) mice were administered intranasally a medium (the same emulsion without DCE), a DCE emulsion (KO-4.4s), and a composition of a particulate emulsion of DCE according to the invention (KO-510), in which the formation of DCE particles was limited by a high concentration of surfactant (polysorbate 80) that also promoted their dissolution.

[0198]

[0199] In short, the formulations KO-4.4s and KO-510 contain 40 and 38 mg / ml of DCE, respectively, and 10 μL of the formulation is delivered to each nostril once daily for one month.

[0200] DCE and its active metabolite creatine were then detected in one hemisphere of each mouse by LC / MS / MS. Results are reported in Figure 3-4 middle.

[0201] In mice treated with the granular emulsion (i.e., KO-510) according to the invention for spraying, high concentrations of DCE and creatine were observed in the brain. This establishes that the composition in granular emulsion form according to the invention delivers DCE and its active metabolite creatine more effectively to the brain, particularly via nasal spray.

[0202] Furthermore, safety studies in animals have shown that while the granular emulsion according to the invention administered nasally is safe, i.e. well tolerated by the nasal mucosa (6 mg / kg / day in rats and 1 mg / kg / day in dogs, both administered nasally for 30 days), the formulation KO-4.4s induced local toxicity in the nasal cavity (0.9 to 2.3 mg / kg / day in cynomolgus monkeys for 30 days) due to histopathological studies showing the presence of cellular inflammation, necrosis, and cellular debris and / or exudates.

Claims

1. A composition in the form of a particulate emulsion, said composition comprising: - At least one creatine ester or creatine ester salt in particulate form; -50% (w / w) to 80% (w / w) of aqueous solvents; -10% (w / w) to 40% (w / w) of lipids.

2. The composition according to claim 1, wherein the composition does not contain ω-3 fatty acids.

3. The composition according to claim 1 or 2, further comprising at least one wetting agent.

4. The composition according to any one of claims 1 to 3, further comprising at least one stabilizer.

5. The composition according to any one of claims 1 to 4, further comprising at least one surfactant, particularly at least one nonionic surfactant, and more particularly, wherein the concentration of said at least one surfactant is less than 15% (w / w).

6. The composition according to any one of claims 1 to 5, further comprising sodium chloride (NaCl).

7. The composition according to any one of claims 1 to 6, wherein, The aqueous solvent is water.

8. The composition according to any one of claims 1 to 7, wherein the lipid comprises triglycerides, particularly medium-chain triglycerides, or is composed of triglycerides, particularly medium-chain triglycerides.

9. The composition according to any one of claims 1 to 8, wherein the creatine has the following formula (I): NH2-C(NH)-N(CH3)-CH2-COOR (I) Where R represents: - Hexosyl, which is optionally substituted by at least one group selected from the list of the following: alkyl containing 1 to 5 carbon atoms, aryl containing 5 to 10 carbon atoms, alkoxy containing 1 to 5 carbon atoms, halogen atom, cyano, trifluoromethyl or nitro. - An alkyl group comprising 1 to 30 carbon atoms, wherein the alkyl group is optionally substituted with at least one group selected from the following: an alkyl group comprising 1 to 5 carbon atoms, an aryl group comprising 5 to 10 carbon atoms, an alkoxy group comprising 1 to 5 carbon atoms, a halogen atom, a cyano group, a trifluoromethyl group, or a nitro group. - An alkenyl group comprising 2 to 30 carbon atoms, optionally substituted with at least one group selected from: an alkyl group comprising 1 to 5 carbon atoms, an aryl group comprising 5 to 10 carbon atoms, an alkoxy group comprising 1 to 5 carbon atoms, a halogen atom, a cyano group, a trifluoromethyl group, or a nitro group; or - An aryl group comprising 5 to 30 carbon atoms, wherein the aryl group is optionally substituted by at least one group selected from the following: an alkyl group comprising 1 to 5 carbon atoms, an aryl group comprising 5 to 10 carbon atoms, an alkoxy group comprising 1 to 5 carbon atoms, a halogen atom, a cyano group, a trifluoromethyl group, or a nitro group.

10. The composition according to any one of claims 1 to 9, wherein the creatine ester is dodecyl creatine ester (DCE) and / or nonyl creatine ester (NCE).

11. The composition according to any one of claims 1 to 10, wherein the creatine ester or creatine ester salt is micronized.

12. The composition according to any one of claims 1 to 11, comprising 0.1% (w / w) to 10% (w / w) of the creatine ester or creatine ester salt.

13. The composition according to any one of claims 1 to 12, which is suitable for spray application.

14. The composition according to any one of claims 1 to 13, wherein it is a pharmaceutical composition or a medical device.

15. The composition according to any one of claims 1 to 14, used as a medicine.

16. The composition according to any one of claims 1 to 14, for the prevention or treatment of creatine deficiency; mitochondrial diseases, such as Leich syndrome, Leber hereditary optic neuropathy, or MELAS syndrome; neurodegenerative diseases, particularly Parkinson's disease and related diseases, or multiple system atrophy (MSA); neuromuscular disorders; hypoxia; ischemic encephalopathy, such as stroke; cardiovascular diseases; muscular dystrophy; skin diseases, such as atopic dermatitis or skin lesions; amyotrophic lateral sclerosis; adrenoleukodystrophy; leukodystrophy; Rett syndrome; Kabuki syndrome; or inflammation.

17. The composition according to claim 16, for the prevention or treatment of creatine transporter deficiency (CTD).

18. A method for preparing a composition in the form of a particulate emulsion, said composition comprising at least one particulate creatine ester or creatine ester salt, said method comprising: - A solution of at least one surfactant is added to a first mixture of at least one creatine ester or creatine ester salt and lipid to form a second mixture; - Stir the second mixture to form a particulate emulsion comprising at least one particulate form of creatine ester or creatine ester salt. The concentration of the at least one surfactant in the composition in particulate emulsion form is less than 15% (w / w).

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