阅读说明：本技术 用于治疗乙酰胆碱依赖性疾病的acefapc (ACEFAPC for the treatment of acetylcholine dependent diseases ) 是由 M·拉加德 E·维瑞塞尔 M·皮克 M·吉康丹 N·伯纳德-赫巴克 B·福莫 于 2018-03-08 设计创作，主要内容包括：本发明涉及用于在预防和治疗与乙酰胆碱缺乏相关联的疾病中使用的AceFaPC(1-乙酰基-2-脂肪酰基-甘油磷酰胆碱)。本发明还涉及该AceFaPC分子和包含该AceFaPC分子的药物组合物,在该AceFaPC分子中Fa表示包含至少14个碳原子的不饱和酰基。(The present invention relates to AceFaPC (1-acetyl-2-fatty acyl-glycerophosphorylcholine) for use in the prevention and treatment of diseases associated with acetylcholine deficiency. The invention also relates to the AceFaPC molecule, wherein Fa represents an unsaturated acyl group comprising at least 14 carbon atoms, and to a pharmaceutical composition comprising the AceFaPC molecule.)

1. AceFaPC of general formula (I), a hydrate, a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate, for use in the prevention and treatment of diseases associated with acetylcholine deficiency,

wherein R represents an acyl group of an unsaturated fatty acid having at least 14 carbon atoms.

2. The AceFaPC according to claim 1, characterised in that the disease associated with acetylcholine deficiency is selected from alzheimer's disease and neuromuscular disease.

3. The AceFaPC according to claim 1 or claim 2, characterized in that it is administered by a suitable route to avoid the gastrointestinal tract.

4. The AceFaPC according to claim 3, characterized in that it is administered by intravenous, intramuscular, subcutaneous, transdermal or inhalation route.

5. Mixture of acefapcs of formula (I), characterized in that it comprises a first AceFaPC of formula (I), wherein R is a first acyl group of an unsaturated fatty acid containing at least 14 carbon atoms, and at least a second AceFaPC of formula (I), wherein R represents an acyl group of an unsaturated fatty acid containing at least 14 atoms, which acyl group is different from the acyl group of the first AceFaPC.

6. AceFaPC mixture according to claim 5, characterized in that it comprises a first AceFaPC of formula (I), wherein R is the acyl group of DHA (AceDoPC), and at least one AceFaPC of formula (I), wherein R represents the acyl group of an unsaturated fatty acid containing at least 14 carbon atoms and is not the acyl group of DHA.

7. Mixture according to claim 5, characterized in that the unsaturated fatty acid of the second AceFaPC of formula (I) is selected from the group consisting of Oleic Acid (OA), Linoleic Acid (LA), alpha-or gamma-linolenic acid (ALA or GLA), eicosapentaenoic acid (EPA) and arachidonic acid (ARA).

8. The AceFaPC mixture of claim 7, wherein the unsaturated fatty acid of the second AceFaPC is arachidonic acid (ARA).

9. AceFaPC mixture according to claim 5, characterized in that the relative proportions of fatty acid acyl groups in the different AceFaPCs of formula (I) are as follows

10. A combination product for simultaneous or time-varying use, characterized in that it comprises, on the one hand, AceDoPC and, on the other hand, at least one AceFaPC of formula (I) wherein R represents the acyl group of an unsaturated fatty acid containing at least 14 carbon atoms and R is not the acyl group of DHA.

11. Pharmaceutical composition, characterized in that it comprises an AceFaPC mixture according to one of claims 5 to 9 and at least one pharmaceutically acceptable excipient.

12. Composition according to claim 11, characterized in that it is in a form suitable for administration by intravenous, intramuscular, subcutaneous, transdermal or inhalation route.

Technical Field

The present invention relates to AceFaPC (1-acetyl-2-fatty acyl-glycerophosphorylcholine) for use in the prevention and treatment of diseases associated with acetylcholine deficiency. The invention also relates to the AceFaPC molecule, wherein Fa represents an unsaturated acyl group containing at least 14 carbon atoms, and to a pharmaceutical composition comprising the AceFaPC molecule.

Prior Art

AceDoPC (1-acetyl, 2-docosahexaenoic acid (docosahexaenoyl) -phosphatidylcholine) is a docosahexaenoic acid (DHA) transporter, well known to the person skilled in the art, the enzymatic synthesis of which is described in application WO 2008/068413. It is known in particular as a modulator of platelet activation of PAF (WO2013037862A 1). AceDoPC has also been shown to promote a reestablished pathway to the blood brain barrier compared to unesterified DHA or PC-DHA (Hashem M. et al, mol. Neurobiol.2016; Bernoud-Hubac N. et al, OCL, 2017). Another study showed that AceDoPC, which acts as a DHA transporter in the brain, can prevent the spread of brain lesions when injected into rats after ischemic stroke (Chauveau et al Curr Neurovasc Res.2011; 8:95-102 and Lagarde M. et al OCL 2016,23(1) D102). Although in vivo studies were conducted on the treatment of ischemia only in rat models, the use of AceDoPC as a supplier of brain DHA was thought to be associated with neurological diseases associated with DHA deficiency (Hachem m. et al, Mol neurobiol.,2016,53(5), 3205-15). The link between DHA and prevention of Alzheimer's disease has been mentioned, in particular the fact that Alzheimer's patients have a deficiency in DHA. Thus, DHA transporters such as AceDoPC are thought to be helpful in preventing Alzheimer's disease but not in treating Alzheimer's disease. When the fatty acid is not DHA, this assumption related to DHA transport cannot generally be extended to AceFaPc molecules.

Although the research has focused on DHA transport and supply, the inventors have now demonstrated that AceDoPC and in general AceFaPC (where Fa is the acyl group of an unsaturated fatty acid of at least 14 carbon atoms) can rapidly transfer its acetyl group to a substrate comprising an alcohol, in particular a primary alcohol.

Based on this observation, the inventors believe that AceDoPC no longer acts as a carrier for DHA, but is the supplier of acetyl groups. Thus, since AceDoPC also contains choline groups, the inventors believe that there is a possibility of producing acetylcholine from AceDoPC in environments with poor choline and/or acetyl sources. AceDoPC can also acetylate the sulfhydryl group of coenzyme a (HSCoA) to produce acetyl-CoA, which is the precursor of acetylcholine in the physiological reaction acetyl-CoA + choline → acetylcholine + HSCoA.

This has been demonstrated by a number of experiments that indicate that AceDoPC and, more generally, AceFaPC can be used to treat diseases associated with acetylcholine deficiency, which treatment is not associated with any DHA supply.

Disclosure of the invention

Accordingly, the present invention relates to AceFaPC of formula (I) for use in the prevention and treatment of diseases associated with acetylcholine deficiency

Wherein R represents an acyl group of an unsaturated fatty acid having at least 14 carbon atoms.

The invention also relates to AceFaPC of formula (I')

Wherein R' represents an acyl group of an unsaturated fatty acid having at least 14 carbon atoms (excluding the acyl group of DHA), a hydrate, a pharmaceutically acceptable salt, or a pharmaceutically acceptable solvate.

The invention also relates to mixtures of AceFaPC and AceDoPC of formula (I '), and pharmaceutical compositions comprising AceFaPC of formula (I ') alone or a mixture of AceFaPC and AceDoPC of formula (I ') and suitable excipients for the administration thereof.

Detailed Description

Accordingly, the present invention relates to AceFaPC of the general formula (I)

Wherein R represents an acyl group of an unsaturated fatty acid having at least 14 carbon atoms, a hydrate, a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate.

It relates to these acefapcs of formula (I) for use in the prevention and treatment, more particularly in the treatment, of diseases associated with acetylcholine deficiency.

The invention also relates to a method for the prevention and treatment of a disease associated with acetylcholine deficiency in a patient, comprising administering to the patient an appropriate dose of AceFaPC of formula (I) or a mixture of acefapcs of formula (I).

The invention is particularly suitable for the prevention and treatment (more particularly treatment) of these diseases in humans. In particular, when a patient is determined to have a deficiency in acetylcholine or may develop such a deficiency, treatment will be used.

Diseases associated with acetylcholine deficiency include

Alzheimer's disease associated with a deficiency of acetylcholine in the brain,

neuromuscular transmitted diseases in which acetylcholine deficiency is recognized, in particular neuromuscular diseases, in particular myopathies deficient in acetylcholine.

By "acetylcholine deficiency" is meant that the amount of acetylcholine measured in an organ of an individual is much lower than would be expected from a normal amount in an individual (or healthy individual) in the absence of such a deficiency. This large reduction compared to healthy individuals results in a metabolic imbalance or dysfunction of the organ.

Accordingly, the present invention relates to acefapcs of formula (I) or acefapcs of formula (I'), or mixtures of acefapcs of formula (I), for use in therapy, more particularly for the treatment of diseases associated with acetylcholine deficiency in patients for which it has previously been determined that there is such a deficiency.

The invention also relates to a method for treating a disease associated with acetylcholine deficiency in a patient, the method comprising

a) Selecting patients for whom a deficiency in acetylcholine has been determined, and

b) administering to the patient an appropriate dose of AceFaPC of formula (I) or AceFaPC of formula (I') or a mixture of AceFaPCs of formula (I).

According to a preferred embodiment of the invention, the AceFaPC must be administered in a manner that is substantially "intact" (intact) when it reaches a target organ that must produce acetylcholine to prevent or compensate for acetylcholine deficiency. By substantially "intact" is meant that a sufficient amount of AceFaPC reaches the organ and is unmodified (particularly modification by acetyl hydrolysis).

It has been observed that acetyl group loss is prone to occur in the gastrointestinal tract of mammals (WO 2017/006047). Thus, a preferred mode of administration would be one adapted to avoid the gastrointestinal tract. These include administration by intravenous, intramuscular, subcutaneous, transdermal and inhalation routes.

For acefapcs of formula (I) wherein R represents an acyl group of an unsaturated fatty acid containing at least 14 carbon atoms, the unsaturated fatty acid containing at least 14 carbon atoms is advantageously a fatty acid of more than 18 carbon atoms and up to more than 22 carbon atoms, in particular fatty acids of 16, 18, 20, 22 and 24 carbon atoms. These unsaturated fatty acids are preferably polyunsaturated. These unsaturated fatty acids are well known to those skilled in the art.

They are in particular selected from palmitoleic acid, oleic acid, Linoleic Acid (LA), alpha-or gamma-linolenic acid (ALA or GLA), arachidonic acid (ARA), eicosapentaenoic acid (EPA), dihomo-gamma-linolenic acid, docosahexaenoic acid (DPA), erucic acid and nervonic acid.

Preferably, the group R of AceFaPC of formula (I) is the acyl group of a polyunsaturated fatty acid selected from oleic acid (OL), Linoleic Acid (LA), alpha-or gamma-linolenic acid (ALA or GLA), arachidonic acid (ARA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Preferably, R of AceFaPc of formula (I) is the acyl group of a polyunsaturated fatty acid selected from arachidonic acid (ARA) and docosahexaenoic acid (DHA). These preferred products are known as aceArPC and AceDoPC, respectively.

For AceFaPC of formula I ', R' is advantageously selected from the acyl groups of the fatty acids defined above, except DHA.

"hydrate" refers to a compound in hydrated form. Examples include hemi-hydrates, mono-hydrates and polyhydrates.

Salts of compounds of formula (I) or formula (Γ) according to the invention include salts with acids or bases, depending on the substituents present. These salts include pharmaceutically acceptable salts such as sodium, potassium and calcium salts.

"solvate" means a form of a compound that is associated with one or more solvent molecules, particularly solvent molecules used during synthesis or purification of the compound, but which are not soluble during purification. The solvent in question will be pharmacologically acceptable.

According to the present invention, "appropriate dose" or "appropriate amount" means any amount that increases the amount of acetylcholine and preferably restores the amount of acetylcholine close to the normal amount expected in a healthy individual. This appropriate dose may be taken once or several times with repeated doses over time. To the extent that the disease being treated is a chronic condition, treatment may be carried out throughout the life of the patient, with appropriate dosages being adjusted according to the progression of the disease.

For the prevention and treatment of diseases associated with acetylcholine deficiency, the skilled person may choose to use AceFaPC alone or a mixture of acefapcs. They may also combine AceFaPC or a mixture of acefapcs with standard treatments for diseases associated with acetylcholine deficiency.

The invention also relates to a mixture of acefapcs of formula (I) as defined above, comprising at least two acefapcs of different all ratios of the groups R. According to a preferred embodiment of the invention, at least one AceFaPC in the mixture is an AceDoPC, which is a molecule of formula I wherein R is an acyl group of DHA. Thus, a preferred mixture according to the invention is a mixture comprising AceDoPC and at least one AceFaPC of formula (I') wherein R is an acyl group of a polyunsaturated fatty acid selected from oleic acid (OL), Linoleic Acid (LA), alpha-or gamma-linolenic acid (ALA or GLA), arachidonic acid (ARA) and eicosapentaenoic acid (EPA), preferably ARA, in all proportions.

The preparation of AceFaPC is known to the person skilled in the art, in particular according to the methods described in applications WO 2008/068413 or WO 2017/006047. The mixture of acefapcs according to the present invention may be prepared by mixing two purified acefapcs or by preparing acefapcs from a source of unsaturated phosphatidylcholine comprising a mixture of unsaturated fatty acids, e.g. a mixture of DHA and ARA in predetermined proportions.

The AceFaPC mixture according to the invention may comprise more than two different acefapcs, especially when the source of unsaturated phosphatidylcholine comprises a mixture of more than two unsaturated fatty acids.

Sources of unsaturated fatty acids useful for preparing acefapcs (particularly mixtures of acefapcs) according to the present invention are well known to those skilled in the art. Examples are egg yolk phosphatidylcholine (2) containing 60% oleoyl (18:1), 30% linoleoyl (18:2), 8% arachidonoyl (20:4) and 2% docosahexenoyl (22:6) at the sn-2 positionhttps://kewpie.co.jp)。

Advantageously, the weight ratio of the first AceFaPC to the second AcFaPC ranges from 1/99-99/1. Especially when the first AceFaPC is AceDoPC, the AceDoPC content in the mixture ranges from 1 to 10 wt% or more, which may depend on the source of the unsaturated phosphatidylcholine and its DHA content, as well as any steps of purification and concentration of the mixture.

The following relative proportions of fatty acid acyl groups will advantageously be present in the AceFaPC mixture according to the invention

According to particular embodiments of the invention, the relative docosahexaenoic acyl content in this mixture may range from 1% to 10%.

The invention also relates to a combination product, or "kit of parts", for simultaneous or time-delayed use, comprising on the one hand an AceDoPC and on the other hand at least one AceFaPC of formula (I) as defined above (which is not an AceDoPC).

The invention also relates to a pharmaceutical composition comprising at least one mixture of acedopcs and at least one AceFaPC of formula (I') (AceFaPC of formula (I), which is not AceDoPC) and at least one pharmaceutically acceptable excipient.

The person skilled in the art is familiar with pharmaceutically acceptable excipients which can be used for the preparation of pharmaceutical compositions, in particular those described in pharmacopoeial standards. They will preferably select excipients that protect the structure of AceFaPC for its storage, in particular to prevent hydrolysis of the sn-1 position leading to loss of acetyl groups.

The pharmaceutical composition is preferably in a form suitable for administration by intravenous, intramuscular, subcutaneous, transdermal or inhalation routes.

Description of the drawings

FIG. 1 depicts the synthesis of acetylcholine from AceFaPC by phospholipase D

Figure 2 shows the detection of acetylcholine and its major fragment products by mass spectrometry.

Figure 3 shows the percentage of passage from blood to brain (rat) based on radioactive DHA as a function of time. (Hashme et al mol. Neurobiol.2016)

Examples

Several experiments were performed using AceFaPC (DHA and oleic acid) as possible precursor of acetylcholine, and the final measurement of AceFaPC was performed by radio-chromatography or mass spectrometry.