Novel hydroxypropyl-β-cyclodextrin and method for producing the same

Abstract

To provide a hydroxypropyl-β-cyclodextrin (HPβCD) having an improved quality, for use as an excipient or as an active pharmaceutical ingredient, as well as to provide the use of the HPβCD as an excipient and a composition comprising the HPβCD as a medicine.SOLUTION: A hydroxypropyl-β-cyclodextrin (HPβCD) has an average degree of molar substitution (MS) of less than or equal to 0.71 and a β-cyclodextrin (β-CD) content of less than or equal to 0.3% by dry weight.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to novel hydroxypropyl-β-cyclodextrin (HPβCD), and to novel methods useful for its preparation. The invention also relates to the use of this HPβCD as an excipient. The invention also relates to HPβCD for use as a medicament, more particularly for the treatment or prevention of diseases or conditions associated with cholesterol overload, and / or storage and / or accumulation within tissues, and thus as a result thereof, for example certain disorders of the central nervous system or the cardiovascular system.

Background Art

[0002] Cyclodextrins are cyclic oligosaccharides derived from the enzymatic degradation of starch. The three most common natural cyclodextrins are composed of 6, 7 or 8 α-D-glucopyranose units linked to each other by α-1,4 bonds in a chair conformation. They are each more commonly referred to as α, β, or γ-cyclodextrin. The appearance of their tertiary structure is in the form of a truncated cone, with its outer side being hydroxyl groups that represent the highly hydrophilic part of the cyclodextrin. The interior or cavity of the cyclodextrin cone is rendered nonpolar since it is composed of hydrogen atoms by C3 and C5 carbons and oxygen atoms further involved in the glycosidic bond.

[0003] Cyclodextrins having a hydrophilic outer part and a hydrophobic cavity are generally used as their ability to encapsulate lipophilic compounds or groups, and thus their role as protectors and solubilizers of these lipophilic compounds or compounds having lipophilic groups. Therefore, they are usually found not only in the field of food processing but also in galenicals, where they are used as excipients in pharmaceutical formulations.

[0004] Each anhydrous glucose unit of cyclodextrin contains three reactive hydroxyl groups derived from carbons C2, C3, and C6. Therefore, by grafting different groups onto these hydroxyl groups, a great many derivatives have already been synthesized, among which hydroxypropyl cyclodextrin, methyl cyclodextrin, and sulfoalkyl cyclodextrin are sometimes mentioned.

[0005] In particular, hydroxypropyl-β-cyclodextrin (HPβCD) is widely used in galenus for oral or parenteral delivery of the active ingredient.

[0006] Typically, HPβCD is obtained by reacting β-cyclodextrin with propylene oxide in a basic medium, after which the reaction product is neutralized by adding hydrochloric acid.

[0007] Next, the crude reaction product contains impurities, i.e., typically: - Reagents, e.g., residual (unsubstituted) β-cyclodextrin; - Reaction by-products such as salts, propylene glycol (propane-1,2-diol), and dipropylene glycol; - Decomposition products of β-cyclodextrin or their linear structures; - Microbiological products such as endotoxins It is purified to reduce its content.

[0008] This purification is carried out using one or more processes such as filtration, decolorization, demineralization, washing with ethanol, extraction with acetone, and dialysis.

[0009] From a regulatory standpoint, HPβCD requires a high degree of purity, especially if it is intended for the pharmaceutical market. These regulatory requirements are further heightened if the route of administration is invasive.

[0010] In particular, HPβCD is known to have favorable effects on certain central nervous system (CNS) disorders, such as Niemann-Pick disease type C and other specific intractable diseases. In these conditions, the subarachnoid pathway is preferred, and safety requirements are very high.

[0011] Therefore, preferably, HPβCD should have an average molar degree of substitution (MS) equal to a maximum of 0.71, preferably in the range of 0.50 to 0.71, and should contain as few impurities as possible. Among these impurities, residual β-cyclodextrin (β-CD) has been identified as potentially exhibiting toxic properties.

[0012] A notable technical challenge lies in matching this MS range with a low content of residual β-CD. Naturally, this MS range is particularly low. Preparing HPβCD with a low MS usually means that less hydroxypropylating reagent is used. Consequently, the reaction product will contain more unreacted β-CD, i.e., residual β-CD.

[0013] However, while this β-CD is difficult to remove, a sufficient reduction in its content leads to an increase in MS, because the latter is determined based on all cyclodextrin molecules, including unsubstituted β-CD.

[0014] Furthermore, after hydroxypropylation, it is necessary to reduce reaction by-products such as propylene glycol or dipropylene glycol. This also leads to an increase in the proportion of β-CD, as the total dry mass of HPβCD decreases due to the removal of these other by-products. After hydroxypropylation, the propylene glycol content is typically, for example, at least 5.0% by dry weight.

[0015] Therefore, producing HPβCD with a predetermined MS and predetermined β-CD content is difficult, especially considering that the MS must be equal to a maximum of 0.71 and the β-CD content must be as low as possible. This difficulty increases if it is also desired to minimize by-products such as propylene glycol and dipropylene glycol.

[0016] International Publication No. 2016 / 201137 (VTESSE) proposes various methods for purifying HPβCD, including complex formation with organic compounds, precipitation, and adsorption chromatography on alumina. In this patent application, HPβCD with a reduced β-CD content and an MS of less than 0.71 was obtained. However, all purification processes allow for the acquisition of the organic solvent used, such as acetone or methanol.

[0017] The use of these organic solvents is particularly restricted, and strict supervision is required insofar as they are used, especially to ensure the safety of handlers. Furthermore, these solvents can be found as impurities in the final product, posing a toxicity risk to consumers.

[0018] As a result, the demand for HPβCD with improved safety, particularly with a low MS of 0.71 or less, and especially when the latter is intended to be injected, for example, intravenously or subarachnoidally, has not been met. [Overview of the Initiative] [Problems that the invention aims to solve]

[0019] One object of the present invention is to provide HPβCD having improved quality, particularly for use as an excipient or active pharmaceutical ingredient.

[0020] Another object of the present invention is to provide HPβCD which is particularly effective for solubilizing or stabilizing active drugs, especially active pharmaceutical components.

[0021] Another object of the present invention is to provide HPβCD having a reduced content of residual β-CD, which may contain no undesirable organic solvents such as methanol, ethanol, acetone, acetonitrile and chloroform, as opposed to the prior art case, and having an MS equal to at most 0.71.

Means for Solving the Problems

[0022] After extensive testing, the applicant has achieved this by arriving at the development of HPβCD having an MS equal to at most 0.71 and a β-CD content of at most 0.3% (dry / dry), preferably 0.2% (dry / dry) or less.

[0023] Furthermore, the HPβCD according to the present invention may have remarkable application properties, particularly with respect to the stabilization of pharmaceutically active proteins.

[0024] This HPβCD, unlike the HPβCD used in International Publication No. WO 2016 / 201137 of the patent application, may advantageously not contain the organic solvents normally resulting from the HPβCD purification process.

[0025] In fact, the HPβCD of the present invention may advantageously be obtained by a novel process carried out in an aqueous medium. In the novel method, the solution is present more in the hydroxypropylation step than in the purification step. In particular, the applicant has shown that it is possible to obtain the HPβCD of the present invention by a judicious choice of hydroxypropylation parameters.

[0026] Thus, it is possible to formulate using a purification step that necessarily finds undesirable organic solvents in the final product. The purification step carried out in an aqueous medium is sufficient. The HPβCD thus obtained has an MS within the range of 0.71 or less, particularly 0.50 to 0.71. Its β-CD content is reduced and is 0.3% or less, preferably 0.2% or less. Undesirable reaction by-products such as propylene glycol and dipropylene glycol can also be reduced.

[0027] Therefore, the first subject of the present invention is - having an average degree of molar substitution (MS) of 0.71 or less; and - having a content of β-cyclodextrin (β-CD) of 0.3% or less, preferably 0.2% or less, by dry weight, which is hydroxypropyl-β-cyclodextrin (HPβCD).

[0028] Another subject of the present invention is step (a) of preparing an aqueous solution containing β-cyclodextrin (β-CD) and sodium hydroxide, wherein the amount of sodium hydroxide used is less than 3.7% by dry weight of sodium hydroxide with respect to the dry weight of β-CD; step (b) of adding propylene oxide to the solution obtained in step (a), - the temperature of the solution obtained in step (a) before the introduction of propylene oxide is selected within the range of 80°C to 120°C; - the molar ratio of propylene oxide / glucose anhydrous used is selected within the range of 0.70 / 1.00 to 0.86 / 1.00; - the addition rate of propylene oxide is selected within the range of 0.15 to 0.30 kg / h / kg of β-CD; characterized step (b); and <� purification step (c) characterized by not using an organic solvent; and step (d) of recovering the HPβCD thus obtained, A method for preparing HPβCD, which is particularly useful for the preparation of HPβCD according to the present invention, characterized by comprising the above steps.

[0029] Another subject of the present invention is HPβCD obtained by the method for preparing HPβCD of the present invention.

[0030] The present invention also relates to the use of HPβCD according to the present invention as a drug and / or excipient, and / or for encapsulating a substance, and / or for solubilizing a substance in an aqueous medium, and / or for improving the chemical stability of a substance, and / or for improving the delivery of a substance to and through biological membranes, and / or for enhancing the physical stability of a substance, and / or for formulating a substance from liquid form to powder form, and / or for inhibiting the interaction of one substance with another, and / or for reducing the local irritant effect of a substance after topical or oral administration, and / or for inhibiting the absorption of a substance in specific tissues such as skin, and / or for obtaining sustained release of a substance, and / or for masking the taste of a substance, particularly its bitterness, and / or for masking the odor of a substance, and / or for improving the bioavailability of a substance. [Brief explanation of the drawing]

[0031] [Figure 1] We present a monograph of hydroxypropyl beta-dex (HPβCD) from USP41 NF36, which is an integral part of the description of the patent application. [Figure 2] We present a monograph of hydroxypropyl beta-dex (HPβCD) from USP41 NF36, which is an integral part of the description of the patent application. [Figure 3] We present a monograph of hydroxypropyl beta-dex (HPβCD) from USP41 NF36, which is an integral part of the description of the patent application. [Modes for carrying out the invention]

[0032] Therefore, the present invention relates to hydroxypropyl-β-cyclodextrin (HPβCD), characterized by having an average molar degree of substitution (MS) of 0.71 or less and a β-cyclodextrin (β-CD) content of 0.3% or less by dry weight.

[0033] It is generally understood that the expression "HPβCD" usually encompasses mixtures of HPβCD molecules, and even substances obtained from methods of preparing them. In fact, contrary to chemical substances with clearly defined structures, HPβCD generally refers to mixtures of HPβCD molecules that have different substitution profiles and patterns, and are therefore structurally different.

[0034] First, the HPβCD of the present invention is characterized by its mean molar degree of substitution (MS) of 0.71 or less. Preferably, this MS is selected within the range of 0.50 to 0.71. It is more preferably 0.70 or less, and preferably 0.69 or less. Preferably, this MS is at least equal to 0.58, i.e., selected within the range of 0.58 to 0.71. Furthermore, this MS is generally at least equal to 0.60, and moreover at least equal to 0.65. It is typically equal to 0.66, or equal to 0.67, or equal to 0.68, or equal to 0.69.

[0035] Here, it is worth recalling that the "average molar substitution (MS)" corresponds to the average number of hydroxypropyl groups per unit of anhydrous glucose. It should be noted that MS differs from the average molar substitution (DS), which corresponds to the average number of hydroxypropyl groups per molecule of cyclodextrin, and is therefore a function of the number of anhydrous glucose units constituting the starting cyclodextrin. Thus, in the case of HPβCD, since β-cyclodextrin consists of 7 anhydrous glucose units, DS is equal to 7 times MS.

[0036] MS can usually be determined by those skilled in the art by proton nuclear magnetic resonance (NMR), preferably following the "hydroxypropyl beta-dex molar substitution" method for USP41 NF36, as reproduced in the appendix.

[0037] As described above, this mixture typically contains residual natural β-cyclodextrin (β-CD) molecules, i.e., molecules that are not hydroxypropylated but are advantageously reduced in the HPβCD of the present invention.

[0038] Therefore, the HPβCD of the present invention has a β-CD content of 0.3% or less, preferably 0.2% or less, and preferably 0.1% or less, and this percentage is expressed as the dry weight of β-CD relative to the total dry weight of HPβCD.

[0039] This residual β-CD content can usually be determined by those skilled in the art by high-performance liquid chromatography (HPLC), preferably following the procedure of USP41 NF36 ("Hydroxypropyl beta-dex; limits of beta-dex, propylene glycol, and other related substances") as reproduced in the appendix.

[0040] Preferably, the substitution pattern of HPβCD according to the present invention is as follows: The proportion of the unsubstituted (no-OHP) portion is: - At least equal to 25.0%, preferably 30.0%, preferably 35.0%, preferably 40.0%; and / or - At most equal to 55.0%, preferably 52.0%, preferably 50.0%, preferably 49.0%, preferably 48.0%; and / or, The proportion of the C2 substitution (2OHP) portion is: At least 15.0%, preferably 20.0%, preferably 24.0%, preferably 25.0%, preferably 26.0%, preferably 27.0%, preferably 28.0%, preferably equal to 28.5%; and / or, - at most equal to 35.0%, preferably 31.0%, preferably 30.5%; and / or, The proportion of the C3 substitution (3OHP) portion is: - At least equal to 5.0%, preferably 6.0%, more preferably 7.0%; and / or - At most equal to 10.0%, preferably 9.0%, preferably 8.0%; and / or The proportion of the C6 substitution (OHP) portion is: - at least equal to 1.0%, and even more than 2.0%; and / or - At most equal to 10.0%, preferably 7.0%, preferably 6.0%, preferably 5.0%, preferably 4.0%; and / or, The proportions of the C2 and C3 substitution (2,3-di-OHP) portions are: - At least equal to 5.0%, preferably 8.0%, preferably 9.0%; and / or, - At most equal to 25.0%, preferably 20.0%, preferably 15.0%, preferably 12.0%, preferably 11.0%, preferably 10.0%; and / or, The proportions of the C2 and C6 substitution (2,6-di-OHP) portions are: - At least equal to 1.0%, preferably equal to 2.0%; and / or, -Maximum equal to 10.0%, preferably 6.0%, preferably 5.0%, preferably 4.0%; and / or The proportion of the two C3 substitution (3,3'-di-OHP) parts is: - At least equal to 0.2%, preferably 0.3%, more preferably 0.5%; and / or - at most equal to 2.0%, preferably 1.5%, preferably 1.0%, preferably 0.8%; and / or The proportions of the C2, C3, and C6 substitution (2,3,6-tri-OHP) portions are: - At least equal to 0.5%, preferably 0.6%, preferably 0.7%, preferably 0.8%; and / or, - At most equal to 4.0%, preferably 3.0%, more preferably 2.0%, preferably 1.5%, and preferably 0.8%.

[0041] These percentages correspond to the percentage of anhydrous glucose units having the type of substitution under consideration. For example, a 2OHP value equal to 30.0% suggests that 30.0 mol% of the anhydrous glucose units in HPβCD are substituted with a hydroxypropyl group at the C2 carbon. As a further example, a 3,3'-di-OHP value equal to 0.4% suggests that 0.4 mol% of the anhydrous glucose units in HPβCD are substituted twice at the C3 carbon (i.e., the C3 carbon has two hydroxypropyl groups). As a final example, a 2,6-di-OHP value equal to 5.0% suggests that 5.0 mol% of the anhydrous glucose units in HPβCD are substituted with hydroxypropyl groups at both the C2 and C6 carbons. The first example refers to a single substitution, while the last two examples refer to a double substitution.

[0042] Preferably, the HPβCD according to the present invention contains less than 5.0%, preferably less than 4.0%, preferably less than 3.0%, preferably less than 2.0%, preferably less than 1.0%, and preferably less than 0.5% of substitutions other than those listed above, such as 3,6-OHP substitutions. More preferably, the HPβCD according to the present invention does not contain any other types of substitutions other than those listed above. The expression "no other types of substitutions" is understood to mean that anhydrous glucose units containing such substitutions cannot be detected in particular by the "Hakomori" method, where the method comprises subjecting the HPβCD to the following sequential steps: permethylation, hydrolysis, reduction, and peracetylation.

[0043] Preferably, the HPβCD substitution pattern according to the present invention is as follows: • The proportion of substitutions equivalent to one substitution is: ○ At least equal to 60%, preferably 70%, preferably 73%, and / or ○ Up to 80%, even 78%, even 77%, even 78%; and / or, The proportion of substitutions equivalent to two substitutions is: ○ at least 15%, moreover 20%, moreover 22%, moreover 23%; and / or, ○ Maximum equal to 40%, preferably 35%, preferably 30%, preferably 28%, preferably 26%, preferably 25%; and / or The proportion of substitutions equivalent to three substitutions is: ○ At least 1%, and even 2%; and / or, ○ Maximum equal to 5%, preferably 4%, preferably 3%, and even more than 2%; and / or The C2 / C6 substitution ratio is: ○ At least equal to 2.0, preferably 3.0, preferably 4.0, preferably 5.0; and / or, ○ Up to 10.0, and even 8.0, and even 7.5; and / or The C2 / C3 substitution ratio is: ○ At least equal to 1.5, preferably 2.0, preferably 2.1, preferably 2.2; and / or, ○ At its maximum, it's equal to 3.0, and even 2.5.

[0044] These substitution patterns can be determined by those skilled in the art, for example, by following a method similar to the "Hakomori" method, typically by subjecting HPβCD to the following sequential steps: permethylation, hydrolysis, reduction, and peracetylation.

[0045] For example, it is possible to follow a method as described in U.S. Patent No. 5,096,893, including Column 6, Examples 9 to Column 7, Example 10, and such method is incorporated by reference. Typically, a method as described in U.S. Patent No. 5,096,893 is as follows: 0.07 mol sodium hydride is added to 20 ml of anhydrous dimethyl sulfoxide under argon, and the mixture is heated at approximately 60°C for 1 hour. Next, 4 g of HPβCD, which has been dried (at 110°C for 3 hours) and dissolved in 15 ml of dimethyl sulfoxide, is added, and the mixture is left under argon with stirring at room temperature for 3 hours. The reaction medium is cooled in an ice bath, and 10 ml, 0.161 mol methyl iodide is added dropwise. After another hour in the ice bath, the mixture is left overnight with stirring. Next, 24 ml of water is added while cooling, while the product is extracted twice with chloroform (90 ml total). The extract is washed with water (20 ml) and evaporated. The residue is treated with water (25 ml) and extracted three times with ether (75 ml total). The extract is washed with water and then evaporated. The residue is dissolved in ether (100 ml), then stirred for 30 minutes in the presence of neutral alumina, filtered, and then evaporated until typically 3.7 g of permethylation product is obtained. 3 mg of the permethylation product is dissolved in aqueous trifluoroacetic acid (0.5 ml), then concentrated in a screw-cap tube, stored overnight at 100°C, and rinsed with air. The residue and sodium borohydride (100 mg) are dissolved in aqueous ammonia (0.5 ml), and the solution is left at room temperature for 1 hour. The solution is acidified with 50% acetic acid (2 drops) and then concentrated. Boric acid is evaporated by co-distillation, first with an acetic acid / methanol mixture (1:9, 5 ml) and then with methanol (25 ml). The residue is treated with anhydride acetic acid and pyridine (2:1, 0.5 ml) at 100°C for 30 minutes, concentrated, and separated between chloroform and water (2:1, 6 ml). The chloroform phase is concentrated, and the residue is analyzed by gas-liquid chromatography (GLC-MS) combined with gas chromatography and mass spectrometry. Gas-liquid chromatography is performed, for example, on a Hewlett Packard 5830A instrument equipped with a flame ionization detector using hydrogen as the carrier gas. Gas-liquid chromatography combined with mass spectrometry is performed, for example, on a Hewlett Packard 5790-5970 system using helium as the carrier gas.For example, a Hewlett Packard Ultra 2 type capillary column made of (crosslinked 5% methylphenyl silicone) fused glass (25 m in length, 0.20 mm in inner diameter) is used. The temperature is programmed as follows: 185°C for 8 minutes, increasing to 250°C at 5°C / min, and then to 250°C for 10 minutes.

[0046] The inventors have found that the performance of HPβCD is enhanced when it has a specific substitution pattern. More specifically, the inventors have shown improved stabilization of proteins, particularly therapeutic proteins (hormones, antibodies, etc.).

[0047] Preferably, the HPβCD according to the present invention has the following organic solvent content: - d-limonene in an amount less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; - Ethanol in an amount of less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; - Methanol in a concentration of less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; - Acetonitrile in an amount of less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; - Acetone in a concentration of less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; - Chloroform in an amount of less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; The content of these is expressed as the dry weight of the organic solvent relative to the total dry weight of HPβCD.

[0048] These organic solvent contents can usually be determined by those skilled in the art by gas chromatography (GC / MS) with detection by mass spectrometry, preferably by headspace GC / MS. For example, this can be done by following the method described in Section A of the Examples.

[0049] Most preferably, the HPβCD according to the present invention does not contain d-limonene and / or ethanol and / or methanol, and / or acetonitrile and / or acetone and / or chloroform in any detectable amount.

[0050] Preferably, the HPβCD according to the present invention also has the following organic solvent content: - p-xylene in amounts less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; and / or - Toluene in amounts less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; and / or - l-menthol in an amount less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm; and / or - Trichloroethylene in an amount of less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm, preferably less than 5 ppm, preferably less than 1 ppm; It holds.

[0051] Most preferably, the HPβCD of the present invention does not contain p-xylene and / or toluene and / or l-menthol and / or trichloroethylene in detectable amounts.

[0052] Preferably, the HPβCD according to the present invention is less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm, as per the General Chapter of USP41 NF36 <467> It has a total content of Class 1 organic solvents according to the "Residual Solvent" section.

[0053] Preferably, the HPβCD according to the present invention is less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm, as per the General Chapter of USP41 NF36 <467> It has the total content of a Class 2 organic solvent according to the "Residual Solvent" section.

[0054] Preferably, the HPβCD according to the present invention is less than 2000 ppm, preferably less than 1000 ppm, preferably less than 500 ppm, preferably less than 200 ppm, preferably less than 100 ppm, preferably less than 50 ppm, preferably less than 10 ppm, as per the General Chapter of USP41 NF36 <467> It has the total content of a Class 3 organic solvent according to the "Residual Solvent" section.

[0055] Alternatively, or in addition to characterization by organic solvent content, HPβCD may be defined by the fact that it is available or obtainable by a hydroxypropylation and purification process that excludes the use of organic solvents, i.e., is carried out entirely in an aqueous medium.

[0056] Preferably, the HPβCD according to the present invention has the following substitution profile, as determined by electrospray ionization-mass spectrometry (ESI-MS): - Signal corresponding to unsubstituted β-CD (HP0): equal to 0.0%; and / or - Signal (HP1) corresponding to an HPβCD molecule with a degree of substitution equal to 1: 3% or less, preferably 1% or less, preferably equal to 0%; and / or - Signal (HP2) corresponding to an HPβCD molecule with a degree of substitution equal to 2: 5% or less, preferably in the range of 0-4%, preferably in the range of 1-3%, and / or - Signal (HP3) corresponding to an HPβCD molecule with a degree of substitution equal to 3: within the range of 1-10%, preferably 2-8%, preferably 3-7%; and / or - Signal (HP4) corresponding to an HPβCD molecule having a degree of substitution equal to 4: within the range of 5-20%, preferably 7-17%, preferably 9-15%; and / or - Signal (HP5) corresponding to an HPβCD molecule with a degree of substitution equal to 5: within the range of 10-30%, preferably 15-25%, preferably 17-25%; and / or - Signal (HP6) corresponding to an HPβCD molecule with a degree of substitution equal to 6: within the range of 15-35%, preferably 20-30%, preferably 24.0-28.0%; and / or - Signal (HP7) corresponding to an HPβCD molecule with a degree of substitution equal to 7: within the range of 10-30%, preferably 15-25%, preferably 17-25%; and / or - Signal (HP8) corresponding to an HPβCD molecule with a degree of substitution equal to 8: within the range of 5-20%, preferably 5-15%, preferably 6-13%; and / or - Signal (HP9) corresponding to an HPβCD molecule with a degree of substitution equal to 9: within the range of 1 to 10%, preferably 2 to 8%, preferably 2 to 6%; and / or - Signal (HP10) corresponding to HPβCD molecules with a degree of substitution equal to 10: less than 5%, preferably less than 2%, preferably 1% or less, preferably equal to 1%; and / or - Signals corresponding to HPβCD molecules with a degree of substitution of 11 or more (HP≧11): 2% or less, preferably 1% or less, preferably equal to 0%; These percentages are expressed as the sum of the signals obtained at each degree of substitution, where the signal was larger than that of background noise.

[0057] Naturally, it is accepted that the term "signal" refers to the area under the curve of the ion corresponding to the desired degree of substitution.

[0058] In this specification, the substitution profile is determined by ESI-MS, preferably by taking the average of measurements taken in three different ways. In determining this substitution profile, it is possible to follow the method described below, particularly in Section A of the Examples.

[0059] Preferably, the HPβCD according to the present invention has a propylene glycol content of 5.00% or less, where this percentage is expressed as the dry weight of propylene glycol relative to the total dry weight of HPβCD. This propylene glycol content is preferably 2.50% or less, preferably 1.00% or less, preferably 0.50% or less, preferably 0.10% or less, and preferably 0.05% or less.

[0060] This propylene glycol content can usually be determined by those skilled in the art by high-performance liquid chromatography (HPLC), following the procedure of USP41 NF36 ("Monograph of Hydroxypropyl Betadex; Limits of Betadex, Propylene Glycol, and Other Related Substances"), which is reproduced in the appendix.

[0061] Preferably, the HPβCD according to the present invention has a dipropylene glycol content of 0.10% or less, where this percentage is expressed as the dry weight of dipropylene glycol relative to the total dry weight of HPβCD. This dipropylene glycol content is preferably 0.05% or less, and more preferably 0.03% or less. It is, for example, in the range of 0.01 to 0.05%.

[0062] Preferably, the HPβCD according to the present invention has a reducing sugar content of 1.0% or less, and this percentage is expressed as the dry weight of the reducing sugar relative to the total dry weight of the HPβCD. Preferably, this reducing sugar content is 0.5% or less, and more preferably 0.1% or less.

[0063] This reducing sugar content can usually be determined by those skilled in the art by the Bertrand method, for example, according to the method described in Section A of the Examples below.

[0064] Generally, advantageously, the HPβCD according to the present invention has a chloride content of 1000 ppm or less, which is expressed as the dry weight of chloride ions relative to the total dry weight of HPβCD. Preferably, this chloride content is 500 ppm or less, preferably 100 ppm or less, preferably 50 ppm or less, and preferably less than 50 ppm.

[0065] This chloride content can usually be determined by those skilled in the art by potentiometric titration of the HPβCD solution with a known concentration of silver nitrate.

[0066] Preferably, the HPβCD according to the present invention has a maximum absorbance of 1.00 or less at 230 to 400 nm; the maximum absorbance is measured using a cell with an optical path length of 10 mm based on a distilled aqueous solution containing 2.50 g (dry) of HPβCD per 100 ml of solution. Preferably, this maximum absorbance is less than 0.50, preferably less than 0.10, preferably less than 0.05, and for example, in the range of 0.01 to 0.50.

[0067] In an advantageous embodiment, the HPβCD of the present invention is in powder form. In this case, it advantageously exhibits a dry mass defect (or "moisture content") of 10.0% or less, preferably 5.0% or less, selected from a range such as 2.0 to 5.0%.

[0068] This water content can usually be determined by those skilled in the art by measuring the loss of mass at dryness, following the procedure of USP41 NF36 ("Monograph of Hydroxypropyl Beta Dex; Loss at Dryness"), which is preferably reproduced in the appendix.

[0069] This powder form is particularly advantageous in the storage and transport of HPβCD.

[0070] Advantageously, this powdered HPβCD is in the form of a micronized product, i.e., a powder obtained by spray-drying a solution of HPβCD.

[0071] Preferably, the HPβCD according to the present invention has a pH in the range of 5.0 to 7.5; the pH is measured based on a solution of HPβCD consisting of 2 g of dried HPβCD, 98 g of distilled water, and 0.3 ml of a 225 g / L potassium chloride solution.

[0072] Preferably, the HPβCD according to the present invention has a conductivity of 200 μS / cm or less, and the conductivity is measured based on a distilled aqueous solution containing 10% (dry) HPβCD. Preferably, this conductivity is 100 μS / cm or less, preferably 50 μS / cm or less, preferably 25 μS / cm or less, and preferably 10 μS / cm or less. This conductivity is, for example, in the range of 1 to 10 μS / cm, and more specifically, 2 to 5 μS / cm.

[0073] This conductivity can usually be determined by those skilled in the art by following the procedure described in USP41 NF36 ("Monograph of Hydroxypropyl Beta Dex; Conductivity"), which is reproduced in the appendix. For example, it is possible to follow the method described in Section A of the Examples below.

[0074] Preferably, the HPβCD according to the present invention has a content of impurities other than propylene glycol and β-CD that are less than 0.5%, preferably less than 0.1%.

[0075] The content of impurities in HPβCD other than propylene glycol and β-CD can usually be determined by those skilled in the art by high-performance liquid chromatography (HPLC) following the procedure of USP41 NF36 ("Limits of Betadex, Propylene Glycol, and Other Related Substances"), which is reproduced in the Appendix, and it is understood that impurities in HPβCD other than propylene glycol and β-CD correspond to "other related substances" in the said method.

[0076] Preferably, the HPβCD according to the present invention also conforms to the U.S. monograph effective June 1, 2018. Preferably, the HPβCD according to the present invention also conforms to the Chinese monograph effective June 1, 2018. Preferably, the HPβCD according to the present invention also conforms to the European monograph effective June 1, 2018.

[0077] Another subject of the present invention is, Step (a) of preparing an aqueous solution containing β-cyclodextrin (β-CD) and sodium hydroxide, wherein the amount of sodium hydroxide used is less than 3.7% of the dry weight of sodium hydroxide relative to the dry weight of β-CD; In step (b), in which propylene oxide is added to the solution obtained in step (a), - The temperature of the solution obtained in step (a) before the introduction of propylene oxide is selected to be within the range of 80°C to 120°C; - The molar ratio of propylene oxide / anhydrous glucose used is selected within the range of 0.70 / 1.00 to 0.86 / 1.00; - The rate of propylene oxide addition is selected within the range of 0.15 to 0.30 kg / h / kg for β-CD; Step (b) characterized by: A purification step (c) characterized by not using an organic solvent; Step (d) involves recovering the HPβCD obtained in this way, This is a method for preparing HPβCD, which is particularly useful for preparing HPβCD as described above, and is characterized by containing [a specific ingredient / method].

[0078] Preferably, the dry mass of β-CD in the solution of step (a) is selected within the range of 30 to 70% by weight, where this percentage is expressed as the dry weight of β-CD relative to the total weight of the solution. Preferably, this dry mass of β-CD is selected within the range of 40 to 60%, preferably 45 to 55%, and preferably 50 to 55%.

[0079] Preferably, the amount of sodium hydroxide in the solution of step (a) is selected within the range of 0.5 to 3.6% by dry weight of sodium hydroxide relative to the dry weight of β-CD. More preferably, this amount of sodium hydroxide is 1.0% or more, preferably 1.2% or more, preferably 1.3% or more, preferably 1.4% or more, preferably 1.5% or more, further 1.6% or more, further 1.7% or more, further 1.8% or more, further 1.9% or more, further 2.0% or more, further 2.1% or more, further 2.2% or more, further 2.3% or more, further 2.4% or more, further 2.5% or more, further 2.6% or more, further 2.7% or more, further 2.8% or more, and further 2.9% or more. Preferably, this amount of sodium hydroxide is 3.5% or less, preferably 3.4% or less, preferably 3.3% or less, preferably 3.2% or less, preferably 3.1% or less, preferably 3.0% or less, and preferably 2.9% or less.

[0080] Preferably, in step (b), the temperature of the aqueous solution of β-CD before the introduction of propylene oxide is 85°C or higher, preferably 90°C or higher. This temperature is also preferably 110°C or lower, preferably 100°C or lower. It is selected, for example, within the range of 90-100°C, preferably 94-96°C. It is, for example, equal to about 95°C.

[0081] Preferably, in step (b), the molar ratio of propylene oxide / anhydrous glucose used is equal to 0.75 / 1.00 or higher, preferably 0.80 / 1.00 or higher, preferably 0.82 / 1.00 or higher, preferably 0.84 / 1.00 or higher, and preferably 0.85 / 1.00.

[0082] Preferably, in step (b), the rate of addition of propylene oxide is selected within the range of 0.20 to 0.30 kg / h / kg of β-CD, preferably 0.20 to 0.25 kg / h / kg of β-CD, and preferably 0.21 to 0.23 kg / h / kg of β-CD, for example, equal to 0.22 kg / h / kg of β-CD.

[0083] Next, the reaction can be neutralized, for example, by adding hydrochloric acid.

[0084] To carry out step (c), the HPβCD obtained in step (b) can be subjected to one or more treatments typically selected from filtration, nanofiltration, treatment with activated carbon, and demineralization.

[0085] Preferably, this purification is carried out in this order: (b.1) Decolorization; (b.2) filtration; (b.3) Membrane purification This includes the processing performed by [the specified method / system].

[0086] Preferably, the decolorization step (b.1) is carried out using activated carbon treatment, which is typically performed in batch mode. Preferably, this treatment is carried out at 70°C ± 5°C for at least 1 hour.

[0087] Preferably, step (b.2) includes at least one filtration step through a bag filter. Preferably, step (b.2) includes at least one filtration step through a 0.22 μm cartridge filter. Preferably, step (b.2) includes at least one filtration step through a 0.1 μm cartridge filter before filtration, preferably optionally, performed with a 0.22 μm cartridge filter.

[0088] Preferably, the membrane purification step (b.3) is carried out by nanofiltration, preferably using a nanofiltration module equipped with a membrane having a cutoff threshold of less than 800 Da or a nominal retention rate of more than 65% of CaCl2, at a pressure of less than 35 bar and a temperature of more than 45°C.

[0089] Preferably, the purification further comprises a demineralization step (b.4), which advantageously includes passing through a cation exchange column and then an anion exchange column. Preferably, the demineralization further comprises a passage through a mixing bed. Preferably, step (b.4) is carried out such that the resistivity of the product at the outlet is greater than 500,000 Ω·cm.

[0090] Preferably, processes (b.1) and (b.2) are repeated after membrane purification (b.3) or after an optional demineralization process (b.4).

[0091] Advantageously, particularly when it is desired to obtain powdered HPβCD, the method of the present invention includes the step of hydroxypropylating the purified product after the purification and drying the optionally purified product. This drying step can be carried out by any method known to those skilled in the art, typically by evaporation or spray drying, preferably by spray drying.

[0092] This spray drying may be a single-stage or multi-stage spray drying. In the case of multi-stage spray drying, the spray dryer is coupled to a fluidized bed and optionally integrated with a spray drying tower, making it possible to agglomerate the particles formed by spray drying. The latter process is particularly advantageous when it is desired to obtain powder with a larger average diameter or depending on the desired flow for the resulting powder.

[0093] The HPβCD of the present invention can be used in a variety of applications, which are distinguished here from therapeutic and non-therapeutic applications.

[0094] Therefore, the present invention relates, firstly, to HPβCD according to the present invention for use as a drug.

[0095] Preferably, this use is intended for the treatment or prevention of conditions or diseases associated with cholesterol overload in tissues, and / or storage and / or accumulation, and their consequences. This includes, for example, cardiovascular diseases, vascular diseases, obstructive peripheral artery diseases such as atherosclerosis or atherosclerosis-related complications, central nervous system diseases such as Alzheimer's disease, Parkinson's disease, focal segmental glomerulosclerosis, and lysosomal diseases affecting the central nervous system, such as Niemann-Pick disease, e.g., Niemann-Pick disease type A, Niemann-Pick disease type B, or Niemann-Pick disease type C. Atherosclerosis-related complications treated and / or prevented by the use of HPβCD according to the present invention are, non-limitingly, ischemia, e.g., myocardial ischemia, coronary diseases, angina pectoris, acute coronary syndrome, myocardial infarction, mesenteric infarction, stroke, aneurysm, or arteriovenous disease of the lower extremities.

[0096] The HPβCD of the present invention is intended for use in the treatment of Niemann-Pick disease type C, or in the treatment of focal segmental glomerulosclerosis.

[0097] Preferably, the HPβCD of the present invention is intended for administration to humans or animals, preferably humans.

[0098] HPβCD of the present invention can be administered orally, parenterally, cutaneously, or mucous membrane-wise. Parenteral routes include, for example, subcutaneous, intravenous, intramuscular, or intraperitoneal administration, the latter of which are rather prepared for animals. Mucosal routes include, for example, nasal, pulmonary, or rectal mucosa administration. Cutaneous routes include, for example, transdermal routes, particularly via transdermal devices, typically patches. In the case of treatment and / or prevention of central nervous system diseases, subarachnoid or spinal routes may also be used.

[0099] The present invention also relates to the use of HPβCD according to the present invention in the manufacture of agents intended to treat and / or prevent the above-mentioned conditions and diseases. It also relates to a method for treating and / or preventing the above-mentioned conditions and diseases in a subject, comprising administering HPβCD according to the present invention in a therapeutically effective dose.

[0100] HPβCD according to the present invention also has other possible uses, which are selected in particular from the uses typically found in this type of product.

[0101] Accordingly, the present invention also relates to the use of HPβCD according to the present invention as an excipient and / or for encapsulating a substance and / or for solubilizing a substance in an aqueous medium and / or for improving the chemical stability of a substance and / or for improving the delivery of a substance to and through biological membranes and / or for enhancing the physical stability of a substance and / or for formulating a substance from liquid form to powder form and / or for inhibiting the interaction of one substance with another and / or for reducing the local irritant effect of a substance after topical or oral administration and / or for inhibiting the absorption of a substance in specific tissues such as skin and / or for obtaining sustained release of a substance and / or for masking the taste of a substance, particularly its bitterness and / or for masking the odor of a substance and / or for improving the bioavailability of a substance.

[0102] Preferably, these substances are lipophilic compounds or compounds having at least one lipophilic group.

[0103] These lipophilic compounds, or compounds having at least one lipophilic group, may be selected, for example, from compounds that are slightly soluble, very slightly soluble, or substantially insoluble in water at room temperature (15–25°C). “Slightly soluble water-soluble compounds” is usually intended to mean that 100–1000 ml of water is required to dissolve 1 gram of the compound. For “very slightly soluble water-soluble compounds,” this volume of water exceeds 1000 ml, reaching up to 10,000 ml. For “substantially insoluble compounds,” this volume of water exceeds 10,000 ml. In this regard, see [Figures 1, 2, 3] and, in particular, the definition shown in the European Pharmacopeia reference “1.4 Monographs, 07 / 2014:10,000”.

[0104] The substances specified herein may typically be active agents or undesirable substances depending on the selected application. For example, HPβCD of the present invention may be used, for example, in the form of a deodorizing aerosol to mask malodors. They can also be used to extend the effect of flavorings in food compositions or to dissolve and / or stabilize active agents.

[0105] "Active agent" is generally understood to mean any substance of interest, such as a pharmaceutical, animal drug, food, nutritional supplement, cosmetic, or pesticide. Examples of such active agents include active pharmaceutical ingredients, colorants, and fragrances. Preferably, the active agent of the present invention is an active pharmaceutical ingredient intended for human use.

[0106] The active agents useful for the present invention, particularly active pharmaceutical ingredients, can be not only chemical molecules but also "biological" active agents, such as proteins; nucleic acids, e.g., those derived from DNA or RNA; cells; or active ingredients based on or derived from viruses. Examples of preferred active agents in the present invention include therapeutic proteins, e.g., antibodies or hormones.

[0107] The subject matter of the present invention is also a composition comprising HPβCD and at least one other substance according to the present invention.

[0108] Preferably, the other substances are as defined above in the section relating to the use of HPβCD according to the present invention. These are, for example, active agents, preferably active pharmaceutical ingredients, and / or lipophilic compounds or compounds having at least one lipophilic group and / or compounds that are slightly soluble, very slightly soluble, and even substantially insoluble in water at room temperature (15-25°C).

[0109] These other substances may also be selected from commonly used compounds, depending on the use and / or desired form of galenus, provided that the latter does not contradict the properties desired in the present invention. These other substances may be selected from, for example, binders, (super)disintegrants, and lubricants.

[0110] The HPβCDs and compositions containing them according to the present invention may be any Galenic form that is preferred by those skilled in the art, depending on the intended use. They may be, for example, liquid, solid, or semi-fluid forms. They may be, for example, solutions, especially injectable solutions, suspensions, dispersions, emulsions, pellets, granules, films, powders, gels, creams, ointments, pastes, sticks, tablets, hard capsules, soft capsules, osmotic devices, or patches.

[0111] In this invention, when the concentration of a substance in a solution is referred to as a percentage, unless otherwise indicated, it should be noted that the latter usually corresponds to the amount of dry substance in grams per 100 ml of solution.

[0112] When a reference is made to the dry mass ("dry weight") of a substance, it is naturally assumed that this refers to the mass of the anhydrous substance. In other words, this mass excludes any moisture present in the starting material in powder form.

[0113] The present invention will be better understood by the following embodiments, which are intended to be illustrative and non-limiting. [Examples]

[0114] A. Methods used to characterize HPβCD 1. The moisture content (mass loss at dryness) was determined according to the method of USP41 NF36 ("Monograph of Hydroxypropyl Beta Dex; Mass Loss at Dryness") reproduced in the appendix. 2. The reducing sugar content was determined by the Bertrand method, which involves precipitation of cuprous oxide in a reducing medium, filtration on sintered glass, and weighing of the residue. 3. The pH of the solution was determined by measuring the potential difference between two immersed electrodes at 20-25°C. The HPβCD solution consisted of 2 g (dry) of HPβCD, 98 g of distilled water with a resistivity greater than 500,000 Ω·cm, and 0.3 ml of 225 g / L potassium chloride solution. 4. Chloride (Cl - The content was determined by potentiometric titration of HPβCD solution with silver nitrate solution of known concentration. The maximum absorbance in the 5.230–400 nm range was determined using a cell with a light path length of 10 mm, based on a distilled aqueous solution containing 2.50 g (dry) of HPβCD per 100 ml of solution. 6. The content of related substances in HPβCD (β-CD, propylene glycol, and other related impurities (including dipropylene glycol)) was determined according to the method in accordance with USP41 NF36 ("Monograph of Hydroxypropyl Beta-Dex; Limits of Beta-Dex, Propylene Glycol, and Other Related Substances"). 7. Conductivity was measured at 25°C based on a 100 ml solution containing 10% HPβCD prepared in distilled water with a resistivity greater than 500,000 Ω·cm, following the procedure described in USP41 NF36 ("Monograph of Hydroxypropyl Beta Dex; Conductivity") reproduced in the Appendix. The resistivity R of the thus obtained solution was determined by an electronic conductivity meter, and the conductivity was calculated from the latter (1 / R). 8. The mean molar degree of substitution (MS) was determined by NMR according to the method of USP41 NF36 “Monograph of hydroxypropyl betadex; molar substitution” reproduced in the appendix. 9. The substitution profile was determined by electrospray ionization-mass spectrometry (ESI-MS). A 1 g (dry) / L HPβCD solution was prepared in a methanol / water mixture (50 / 50, v / v) with 1 mM sodium acetate. Each sample was injected at 10 μl / min for 1 minute, and the MS data was recorded as follows. Between two consecutive injections, 500 μl of methanol / water mixture (50 / 50, v / v) was injected to wash the ion source. The electrospray ionization (ESI) parameters were as follows: spray voltage: 5kV; nebulizer gas: 9, auxiliary gas: 2; sweep gas: 0; capillary voltage: 23V; capillary temperature: 275℃; tube lens: 80V. The mass spectrometry parameters were as follows: full scan; scan range: 50~200m / z; mass range: normal, scan speed: improved; capture time: 1 minute. For each substituted HPβCD molecule (referred to as HPX, where X is the number of substitutions per β-CD molecule), the extracted ion current (XIC) for each ion was integrated and compared to the sum of all HPX ion currents. The intensity of the corresponding peak was greater than the intensity of the background noise. Since the sodium adduct was the strongest HPβCD ion, the area under the curve of the peak corresponding to each HPX was integrated and related to the sum of the areas of HPX ions considered for characterization (intensity greater than the background noise), and this was expressed as a percentage. 10. The organic solvent content was determined by gas chromatography (GC / MS) with detection by mass spectrometry. More precisely, the analysis was performed by headspace GC / MS. The operating conditions were as follows: Bruker GC / MS with a 30m*0.25mm Vf-wax column with a df of 0.25μm; temperature program: 5 mins at 40°C, increasing to 230°C at 5°C / min; injector at 250°C with a split ratio of 1:10; EI + MS detector. The sample was prepared as follows: 0.2 g of dried HPβCD was dissolved in 1 ml of ultrapure water. 11. The substitution pattern was determined according to the "Hakomori" method discussed above.

[0115] B. Prior Art: MS and β-CD content values ​​of commercially available HPβCD In this section, MS and β-CD content values ​​were measured for various commercially available HPβCDs (Meas.). Supplier details were also provided as information where available (Spec.).

[0116] [Table 1]

[0117] These results confirm that commercially available HPβCD does not correlate with low MS and low residual β-CD content. For example, KLEPTOSE® HP, CAVASOL® W7 HP7, and COMPLEXOL-HP products, although having a β-CD content of 0.3% or less, systematically have an MS exceeding 0.71. Conversely, KLEPTOSE® HPB, CAVASOL® W7 HP, CAVASOL® W7 HP5, and C*CAVITRON® products have an MS of less than 0.71, but their residual β-CD content systematically exceeds 0.3%.

[0118] C. Preparation and characterization of HPβCD (MS and βCD) of the present invention or not (of the present invention) 1. Method 1 - Purification by decolorization, filtration, and membrane purification. The purpose of this section is to present the effect of hydroxypropylation parameters on the properties of the resulting HPβCD.

[0119] HPβCD(IN-1) of the present invention was prepared as follows: 1313 g of commercially available β-cyclodextrin (β-CD) (corresponding to 1188 g of anhydrous β-CD) was dissolved in an alkaline medium while stirring in an autoclave under an inert atmosphere. In this way, a solution containing 52% β-CD by dry weight relative to the total weight of the solution and 2.9% sodium hydroxide ([NaOH] in the table below) relative to the dry weight of β-CD was obtained. The reaction medium was maintained at a temperature of 95°C [T] for 30 minutes, and then 361.6 g of propylene oxide was added at a rate of 0.22 kg / h / kg [D] of dry β-CD, i.e., a propylene oxide / anhydrous glucose [PO / G] molar ratio of 0.85 / 1.00. After the introduction of propylene oxide was completed, the reaction medium was stirred for 4 hours and then neutralized with hydrochloric acid.

[0120] Next, HPβCD was purified and dried in an aqueous medium, i.e., without using an organic solvent, using the following technique: - bleaching; - Filtration; - Membrane purification; - Evaporation drying

[0121] The activated carbon decolorization step was performed in batch mode with stirring at 70°C ± 5°C for a minimum of 1 hour. The medium was then filtered through a cricket filter and then through a 0.22 μm cartridge filter. The membrane purification step was performed using a nanofiltration module with a membrane having a cutoff threshold of 800 Da, particularly at a pressure of less than 35 bar and a temperature of more than 45°C. The propylene glycol content of the retaining solution was observed by HPLC assay. The operation was stopped immediately after the propylene glycol content relative to the dried product reached less than 0.5% by weight. The HPβCD thus obtained was then dried under reduced pressure in a rotary evaporator to have a solid content of more than 95% by weight.

[0122] A comparative sample, HPβCD(CP-1), was prepared as follows: 656 g of commercially available β-cyclodextrin (β-CD) (equivalent to 594.4 g of anhydrous β-CD) was dissolved in an alkaline medium in an autoclave under an inert atmosphere with stirring. In this way, a solution containing 52% β-CD by dry weight relative to the total weight of the solution, and 2.9% sodium hydroxide relative to the dry weight of β-CD was obtained. The reaction medium was maintained at a temperature of 75°C [T] for 30 minutes, and then 182.6 g of propylene oxide was added at a rate of 0.14 kg / h / kg [D] of dry β-CD, i.e., a propylene oxide / anhydrous glucose [PO / G] molar ratio of 0.86 / 1.00. After the introduction of propylene oxide was complete, the reaction medium was stirred for 4 hours and then neutralized with hydrochloric acid. The HPβCD thus obtained was purified and dried in the same manner as HPβCD IN-1.

[0123] The obtained HPβCDs were characterized in relation to their MS and β-CD content. The hydroxypropylation conditions and the values ​​of the MS and β-CD content are shown in the table below.

[0124] [Table 2]

[0125] Next, we conducted the following tests: In each test, approximately 250 g of anhydrous β-CD was used, and the amount of water and propylene oxide introduced was proportional to the amount used in the preparation of HPβCD IN-1. In particular, this meant that the [PO / G] molar ratio was equal to 0.85 / 1.00. The temperature [T] of the reaction medium before the introduction of propylene oxide was 110°C. The various amounts of sodium hydroxide [NaOH] tested are shown in the table below as the dry percentage of sodium hydroxide relative to the dry weight of β-CD. Propylene oxide was added at rate [D]. After the reaction, the reaction medium was neutralized with hydrochloric acid. For each sodium hydroxide [NaOH] content tested, two crude reaction products were produced and then combined to have sufficient material for the nanofiltration step of purification. The thus obtained HPβCD crude reaction products were purified and dried in the same manner as HPβCD IN-1.

[0126] The obtained HPβCDs were characterized in relation to their MS and β-CD content. The hydroxypropylation conditions and the values ​​of the MS and β-CD content are shown in the table below.

[0127] [Table 3]

[0128] All of these tests demonstrate that, when the hydroxypropylation conditions are appropriately designed, it is possible to obtain HPβCD having the MS and β-CD content as the subject of the present invention.

[0129] A significant advantage in purification is that the presence of organic solvents is not required. Therefore, the HPβCD according to the present invention may not contain any organic substances other than those typically obtained from the hydroxypropylation step and the raw materials used in this hydroxypropylation.

[0130] 2. Method 2 - With decolorization and additional filtration The purpose of this section is to present an optimized process for the preparation of HPβCD according to the present invention.

[0131] HPβCD(IN-2) according to the present invention was prepared as follows: 1305 g of commercially available β-cyclodextrin (β-CD) (corresponding to 1204 g of anhydrous β-CD) was dissolved in an alkaline medium while stirring in an autoclave under an inert atmosphere. In this way, a solution was obtained containing 52% β-CD by dry weight relative to the total weight of the solution, and 2.9% sodium hydroxide relative to the dry weight of β-CD. The reaction medium was maintained at a temperature of 95°C [T] for 30 minutes, and then 367 g of propylene oxide was added at a rate of 0.22 kg / h / kg [D] of dry β-CD, i.e., a propylene oxide / anhydrous glucose [PO / G] molar ratio of 0.85 / 1.00. After the introduction of propylene oxide was completed, the reaction medium was stirred for 4 hours and then neutralized with hydrochloric acid.

[0132] Next, the obtained HPβCD was purified and dried in an aqueous medium, i.e., without using an organic solvent, by following the steps below: - bleaching; - Filtration; - Membrane purification; - Demineralization; - bleaching; - Filtration; - spray drying

[0133] The decolorization step was performed by activated carbon treatment at 70°C for 1 hour. The medium was then filtered through a bag filter, then a 1 μm filter, and then a 0.22 μm cartridge filter. The membrane purification step was performed using a nanofiltration module with a membrane having a cutoff threshold of less than 800 Da, at a pressure of less than 35 bar and a temperature of more than 45°C. The propanediol glycol content of the retained solution was observed by HPLC assay. The nanofiltration operation was stopped immediately after the propanediol content relative to the dried product reached less than 0.1% by weight. The demineralization step was performed by passing the solution through a cationic, then anionic ion exchange column, and finally a mixed bed, in particular, to obtain an outlet resistivity of more than 500,000 Ω·cm. The second decolorization step was performed in batch mode by activated carbon treatment with stirring at 70°C for at least 1 hour. The HPβCD solution was then filtered through a bag filter, then a 1 μm filter, and then a 0.22 μm cartridge filter. Next, the HPβCD solution was spray-dried to obtain HPβCD in powder form.

[0134] Next, a thorough characterization of HPβCD IN-2 of the present invention was performed. The results are shown in the table below:

[0135] [Table 4]

[0136] D. Substitution patterns of HPβCD, either present or not present in this invention In this section, the inventors determine the substitution patterns of the HPβCD of the present invention, the substitution patterns of comparative HPβCDs, and the substitution patterns of commercially available HPβCDs. In the case of HPβCD IN-2, several batches were tested.

[0137] The results, expressed as percentages, are shown in Tables 5 and 6. MS values ​​and residual β-CD values ​​are also reported. "Mono": The proportion of substitutions corresponding to monosubstitutions (2OHP, 3OHP, 6OHP); "Di": The proportion of substitutions corresponding to disubstitutions (2,3-di-OHP, 2,6-di-OHP, 3,3'-di-OHP); "Tri": The proportion of substitutions corresponding to the trisubstitution (2,3,6-tri-OHP); "C2 / C6": The ratio of C2 / C6 substitution ((2OHP + 2,3-di-OHP + 2,6-di-OHP + 2,3,6-tri-OHP) / (6OHP + 2,6-di-OHP + 2,3,6-tri-OHP)); "C2 / C3": The ratio of C2 / C3 substitution ((2OHP + 2,3-di-OHP + 2,6-di-OHP + 2,3,6-tri-OHP) / (3OHP + 2,3-di-OHP + 3,3'-di-OHP + 2,3,6-tri-OHP)

[0138] [Table 5]

[0139] [Table 6]

[0140] Note: For all HPβCDs tested, no signals corresponding to other types of substitution were obtained (e.g., for the 3,6-di-OHP type). Therefore, no detectable amounts were obtained for anhydrous glucose units with substitution types other than those shown in Tables 5 and 6.

[0141] These results indicate that HPβCD according to the present invention has a specific substitution pattern, in particular, compared to the CAVASOL® W7 HP Pharma, CAVITRON® W7 HP5 Pharma, and CAVITRON® W7 HP7 Pharma products that were also analyzed.

[0142] Experiments conducted by the inventors (data not shown herein) suggest that substitution patterns, such as those in the case of HPβCD of the present invention, appear to result in improved efficiency in stabilizing certain properties, particularly substances, especially active pharmaceutical components, and more specifically, biologically active agents such as pharmaceutically active proteins. In particular, for the stabilization of therapeutic proteins, HPβCDs having substitution patterns such as those in the case of HPβCD of the present invention showed a significant reduction in protein aggregation compared to those obtained with CAVASOL® W7 HP Pharma, CAVITRON® W7 HP5 Pharma, and CAVITRON® W7 HP7 Pharma products.

Claims

1. - Having an average molar substitution degree (MS) of 0.71 or less, - Contains β-cyclodextrin (β-CD) at a content of 0.3% or less by dry weight, - Hydroxypropyl-β-cyclodextrin (HPβCD) characterized by having the following HPβCD substitution pattern. - The proportion of substitutions equivalent to one substitution is 60% or more and 80% or less. - The proportion of substitutions equivalent to disubstituted compounds is between 15% and 40%. - The proportion of substitutions equivalent to triploat is between 1% and 5%. - The C2 / C6 substitution ratio is 2.0 or more and 10.0 or less. - The C2 / C3 substitution ratio is between 1.5 and 3.

0.

2. The following organic solvent content: - d-limonene less than 2000 ppm; - Ethanol less than 2000 ppm; - Methanol less than 2000 ppm; - Acetonitrile less than 2000 ppm; - Acetone less than 2000 ppm; - Chloroform less than 2000 ppm; The hydroxypropyl-β-cyclodextrin according to claim 1, characterized in that it contains the following, wherein the content of these is expressed as the dry weight of the organic solvent relative to the total dry weight of the hydroxypropyl-β-cyclodextrin.

3. The hydroxypropyl-β-cyclodextrin according to any one of claim 1 or 2, characterized in that the MS is selected within the range of 0.50 to 0.

71.

4. The hydroxypropyl-β-cyclodextrin according to claim 3, characterized in that the MS is selected within the range of 0.58 to 0.

71.

5. A hydroxypropyl-β-cyclodextrin according to any one of claims 1 to 4, characterized in that the β-CD content is 0.2% or less by dry weight.

6. A hydroxypropyl-β-cyclodextrin according to any one of claims 1 to 4, for use as a pharmaceutical agent.

7. Use of hydroxypropyl-β-cyclodextrin according to any one of claims 1 to 4, for use as an excipient and / or for encapsulating a substance and / or for solubilizing a substance in an aqueous medium and / or for improving the chemical stability of a substance and / or for improving the delivery of a substance to and through biological membranes and / or for enhancing the physical stability of a substance and / or for formulating a substance from liquid form to powder form and / or for preventing interaction between one substance and another substance and / or for reducing local irritation after topical or oral administration of a substance and / or for preventing absorption of a substance in the skin and / or for obtaining sustained release of a substance and / or for masking the taste of a substance and / or for masking the odor of a substance and / or for improving the bioavailability of a substance.

8. A composition comprising hydroxypropyl-β-cyclodextrin as described in any one of claims 1 to 4, and at least one other substance.

9. The composition according to claim 8, characterized in that the other substance is an active agent.

10. The composition according to claim 9, characterized in that the active agent is an active pharmaceutical ingredient.

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