Method for preparing l-serine hydrochloride and use thereof

Abstract

The present invention relates to a method for preparing an L-serine hydrochloride and a use of the L-serine hydrochloride prepared by the preparation method. Specifically, the L-serine hydrochloride according to the present invention has increased water solubility and bioavailability and improved storage stability, is convenient for preparing a composition, and has a superior pharmacological profile. The L-serine hydrochloride can be used for preventing, alleviating, or treating neurological, psychiatric, and cognitive disorders.

Description

Method for producing L-serine hydrochloride and its use

[0001] The present invention relates to a method for producing L-serine hydrochloride and a use of L-serine hydrochloride produced by the method.

[0002] In general, when manufacturing pharmaceutical compositions, physicochemical stability and solubility are crucial factors that determine processing costs and the pharmacokinetics of the product. In particular, solution stability and rapid solubility are essential for the manufacture of syrup-type or rapid-disintegrating tablets.

[0003] Active Pharmaceutical Ingredients (APIs) are manufactured and administered as salts or co-crystals to facilitate purification, facilitate the preparation process by obtaining crystalline powders, enhance solubility, and increase bioavailability. The selection of materials used in the manufacture of these salts or co-crystals must be carefully considered based on the amount of API used.

[0004] Publication patent KR101948066B1 relates to a composition for preventing, improving, or treating neurodevelopmental disorders, which contains L-serine as an active ingredient. In the above prior art, the amount of L-serine administered for effective treatment was 200-400 mg / kg, and a relatively large amount of serine was administered twice a day.

[0005] Meanwhile, patent publication KR10-2022-0131186 relates to a syrup, which is the optimal drug formulation that can be consumed by children with autism spectrum disorder. L-serine used in the manufacture of the syrup of the above-mentioned prior document is an amino acid in the form of a free base in a natural state, and according to the above-mentioned prior document, it appears to have a high water solubility of 425 mg / ml, but in reality, it has a low water solubility of about 100 mg / ml due to the use of a sweetener such as sorbitol, which is used to obtain the convenience of taking children's drugs together with the syrup. This means that, for example, the L-serine that a 30 kg pediatric patient needs to take for treatment is about 12,000 mg (30 x 400 mg) per day, and when converted to the amount of syrup, about 120 ml of syrup needs to be consumed. This is a fairly large dose to administer to children with autism spectrum disorder, who are typically sensitive and difficult to control.

[0006] Meanwhile, when manufacturing by reducing the amount of sweetener or raising the temperature to increase solubility, not only does it show low medication compliance due to change in taste, but storage stability is very poor due to precipitation of L-serine when the temperature is lowered, and there is a problem that browning occurs when stored in a solution state for a long period of time.

[0007] In addition, although L-serine hydrochloride is sold by some reagent companies such as Aldrich, there is no known industrial manufacturing process for economical hydrochloride, and only a small amount of L-serine hydrochloride is sold, which is not suitable for medical use.

[0008] Accordingly, in order to solve the above problem, the inventor of the present invention has developed a novel hydrochloride of L-serine and a method for producing the same, which enables the production of a high-content syrup by increasing the solubility, does not discolor even when stored in an aqueous solution for a long period of time, and has an increased half-life and maximum absorption concentration when administered orally, enabling effective treatment or improvement with only one dose per day.

[0009] The object of the present invention is to provide L-serine hydrochloride having increased water solubility and bioavailability, improved storage stability, ease of composition preparation, and superior pharmacological profile, and an economical method for preparing the same.

[0010] Another object of the present invention is to provide a use of L-serine hydrochloride produced by the above production method for preventing, improving or treating neurological, mental and cognitive disorders.

[0011] To achieve the above purpose, the present invention provides a method for producing L-serine hydrochloride comprising the following steps.

[0012] (a) A step of mixing L-serine of the following chemical formula 1 with an organic solvent;

[0013] (b) a step of adjusting the pH of the mixed solution of step (a) with hydrochloric acid diluted in an organic solvent;

[0014] (c) a step of forming a salt by stirring the solution whose pH has been adjusted in step (b); and

[0015] (d) A method for producing L-serine hydrochloride, comprising the step of filtering the salt produced in step (c) and then washing the excess hydrochloric acid with an organic solvent.

[0016] [Chemical Formula 1]

[0017]

[0018] The present invention also provides L-serine hydrochloride of the following chemical formula 2 manufactured by the above manufacturing method.

[0019] [Chemical Formula 2]

[0020]

[0021] The present invention also provides a pharmaceutical composition for preventing or treating neurological, mental and cognitive disorders, comprising the above L-serine hydrochloride as an active ingredient.

[0022] The present invention also provides a health functional food composition for preventing or improving neurological, mental and cognitive disorders, comprising the above L-serine hydrochloride as an active ingredient.

[0023] L-serine hydrochloride according to the present invention has increased water solubility and bioavailability, improved storage stability, is easy to prepare a composition, and exhibits a superior pharmacological profile. In addition, due to the increased half-life and maximum absorption concentration in a composition comprising L-serine hydrochloride according to the present invention, it is possible to effectively prevent, improve, or treat neurological, mental, and cognitive disorders with only one dose per day.

[0024] Figure 1 shows differential scanning calorimeter (DSC) data of L-serine hydrochloride prepared in Example 3.

[0025] Figure 2 is powder X-ray diffraction (PXRD) data of L-serine hydrochloride prepared in Example 3.

[0026] Figure 3 shows differential scanning calorimetry (DSC) data of L-serine.

[0027] Figure 4 is powder X-ray diffraction (PXRD) data of L-serine.

[0028] Figure 5 shows data obtained by titrating L-serine hydrochloride prepared in Example 3 with a silver nitrate solution (ERC: solid line, U: dotted line).

[0029] Figure 6 shows the results of pharmacokinetic evaluation of L-serine hydrochloride prepared in Example 3.

[0030] Figure 7 shows the behavioral experiment process of a VPA (valproic acid)-induced autism model mouse using L-serine hydrochloride prepared in Example 3.

[0031] Figures 8A and 8B show the results of a 3-chamber test that evaluated the social interaction ability of mice. Figure 8A confirms social novelty, and Figure 8B confirms sociability.

[0032] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In general, the nomenclature used herein is well known and commonly used in the art.

[0033] The present invention relates, in one aspect, to a method for producing L-serine hydrochloride.

[0034] Embodiments of the present invention include:

[0035] (a) A step of mixing L-serine of the following chemical formula 1 with an organic solvent;

[0036] (b) a step of adjusting the pH of the mixed solution of step (a) with hydrochloric acid diluted in an organic solvent;

[0037] (c) a step of forming a salt by stirring the solution whose pH has been adjusted in step (b); and

[0038] (d) A step of filtering the salt produced in step (c) and then washing the excess hydrochloric acid with an organic solvent.

[0039] A method for producing L-serine hydrochloride comprising:

[0040] [Chemical Formula 1]

[0041]

[0042] In the present invention, “L-serine” is a type of amino acid that constitutes proteins, is a monoamide of glutamic acid, and means an L-amino acid having the structure of the chemical formula 1.

[0043] In the present invention, step (a) is a step of mixing L-serine with an organic solvent.

[0044] The organic solvent used in the above step (a) may be selected from those known in the art, as long as it does not deviate from the purpose of the present invention. Specifically, the organic solvent may be at least one selected from the group consisting of acetone, methanol, ethanol, isopropanol, butanol, hexanol, ethyl acetate, toluene, methyl-t-butyl ether, and methylene chloride, preferably acetone, methanol, ethanol, or isopropanol, and most preferably isopropanol or ethanol, but is not limited thereto.

[0045] In addition, the organic solvent can be used by mixing about 10-40% of the total amount of solvent used for smooth stirring, but if more than that is used, the reaction yield decreases and it is not economical. The amount of the organic solvent can be about 6-30 times the weight of L-serine used, preferably about 5-20 times, and most preferably 3-6 times, but is not limited thereto. If too much solvent is used, handling during the process is difficult, and the amount of wastewater discarded after the process increases, so it is important to use a minimum amount to ensure economic feasibility. In addition, when the temperature is raised, some L-serine is converted to D-serine, so it is necessary not to raise the temperature above room temperature.

[0046] In the present invention, step (b) is a step of adjusting the pH of the mixed solution of step (a) with hydrochloric acid diluted in an organic solvent.

[0047] If the solution is adjusted to acidity, the solubility in water increases by more than 5 times compared to neutral L-serine, which can significantly reduce the process volume. The pH adjusted in step (b) can be about 1.0-5.0, preferably about 2.0-4.5, and most preferably about 2.5-4.0, but is not limited thereto. On the other hand, if the pH is too low, the amount of acid used increases, which reduces economic efficiency, and there is a risk of corrosion of equipment used in the process, such as reactors or filters made of metal and weak to acid, and there is a possibility of side reactions, such as cross-esterification of additional solvents used, which significantly reduces economic efficiency.

[0048] Step (b) above uses hydrochloric acid diluted in an organic solvent. If concentrated hydrochloric acid is used undiluted, the temperature may rise excessively, causing some L-serine to be converted to D-serine. Therefore, the hydrochloric acid should be diluted with an organic solvent by a factor of about 1-10, preferably about 2-8, and most preferably about 3-7, and slowly added dropwise while maintaining the temperature below room temperature.

[0049] The organic solvent used in the hydrochloric acid dilution may be selected from those known in the art, as long as it does not deviate from the purpose of the present invention. Specifically, the organic solvent may be at least one selected from the group consisting of acetone, methanol, ethanol, isopropanol, butanol, hexanol, ethyl acetate, toluene, methyl-t-butyl ether, and methylene chloride, preferably acetone, methanol, ethanol, or isopropanol, and most preferably isopropanol or ethanol, but is not limited thereto. In addition, it is most preferable to use the same organic solvent as the organic solvent used in step (a), but is not limited thereto.

[0050] In the present invention, step (c) is a step of forming a salt by stirring the solution whose pH has been adjusted in step (b).

[0051] In the above step (c), the stirring time can be set so that the hydrochloride can be sufficiently formed. Specifically, the stirring can be performed for about 2-48 hours, preferably about 3-36 hours, and most preferably about 3-24 hours, but is not limited thereto.

[0052] It is necessary to stir at a temperature below room temperature to prevent the temperature from rising during stirring and the conversion of L-serine to D-serine.

[0053] In the present invention, step (d) is a step of filtering the salt (solid) produced in step (c) and then washing away excess hydrochloric acid with an organic solvent.

[0054] In the above step (d), filtration can be performed using any filtration method possible in industrial production, such as pressure reduction filtration, centrifugation, or filter dryer.

[0055] In addition, the organic solvent used in the washing may be selected from those known in the art, as long as it does not deviate from the purpose of the present invention. Specifically, it may be at least one selected from the group consisting of acetone, methanol, ethanol, isopropanol, butanol, hexanol, ethyl acetate, toluene, methyl-t-butyl ether, methylene chloride, cyclohexane, and pentane, most preferably acetone, methanol, ethanol, or isopropanol, but is not limited thereto. The amount of the organic solvent may be about 1-8 times that of L-serine, preferably about 2-7 times, and most preferably about 3-6 times. At this time, the organic solvent may have a moisture content of 1% or less in order to prevent a decrease in yield.

[0056] In the present invention, after step (d), a step (e) of drying to remove the solvent may be additionally included.

[0057] In the above step (e), the drying temperature is about 20-80°C, preferably 30-70°C, and most preferably 40-60°C, using a vacuum or hot air drying method, but is not limited thereto. However, care must be taken as discoloration may occur if the drying temperature is too high.

[0058] From another aspect, the present invention relates to L-serine hydrochloride of the following chemical formula 2 manufactured by the above manufacturing method.

[0059] [Chemical Formula 2]

[0060]

[0061] According to one embodiment of the present invention, a crystalline L-serine hydrochloride can be formed.

[0062] The above crystal form may exhibit characteristic peaks at diffraction angles (2θ±0.2) of 16.4°, 16.9°, 18.8°, 19.9°, 20.7°, 22.5°, 25.3°, 26.6°, 27.2°, 27.8°, 29.0°, 30.8°, 32.4°, 33.1°, 37.5°, and 38.1° when measured in a powder X-ray diffraction (PXRD) analysis over a scanning range (2θ): 3 to 50°, and a scanning speed: 6 deg / min.

[0063] Additionally, the above crystal form may be characterized by one endothermic peak at about 150°C in a differential scanning calorimeter (DSC) thermogram.

[0064] In another aspect, the present invention relates to a pharmaceutical composition for preventing or treating neurological, mental and cognitive disorders, comprising L-serine hydrochloride produced by the above production method as an active ingredient.

[0065] The term "treatment" as used herein refers to any act of improving or beneficially changing the symptoms of a disease by administering the composition according to the present invention, the term "prevention" refers to any act of suppressing or delaying a disease by administering the composition according to the present invention, and the term "improvement" refers to any act of improving the bad state of a disease by administering or ingesting the composition of the present invention to a subject.

[0066] The disease or condition targeted for "treatment", "prevention" or "improvement" of the present invention may be a neurological, mental and cognitive disorder disease. The neurological, mental and cognitive disorder disease is a general term for diseases with abnormal brain function as a main lesion, and may include, but is not limited to, autism spectrum, bipolar disorder, Tourette's, tics, Prader-Willi, attention deficit hyperactivity disorder (ADHD), Fragile-X syndrome, Rett syndrome, dementia, Alzheimer's, Parkinson's, schizophrenia, depression and memory disorder. For example, the L-serine hydrochloride may exhibit an effect of improving, delaying, preventing or treating autism spectrum, and more specifically, may exhibit an effect of improving, delaying, preventing or treating childhood autism spectrum.

[0067] For the treatment of the diseases or conditions described above, the compositions or preparations described herein may be administered orally. The term "oral" encompasses swallowing, and "oral administration" is used synonymously with oral administration. Oral administration allows the compounds of the present invention to enter the gastrointestinal tract, or, for example, to be absorbed directly into the bloodstream through the mouth, as in buccal or sublingual administration.

[0068] Suitable compositions for oral administration may be in the form of solids, liquids, gels, or powders, and may have dosage forms such as tablets, lozenges, capsules, granules, powders, and liquids.

[0069] Compositions for oral administration may optionally be enteric coated, and may exhibit delayed or sustained release through the enteric coating. That is, the compositions for oral administration according to the present invention may be formulations having an immediate or modified release pattern.

[0070] Liquid formulations may include solutions, syrups, and suspensions, and the liquid compositions may be contained within soft or hard capsules. The formulations may include pharmaceutically acceptable carriers, such as water, ethanol, polyethylene glycol, cellulose, or oil. The formulations may also include one or more emulsifiers and / or suspending agents.

[0071] More specifically, in the present invention, the syrup formulation refers to a concentrated liquid containing sugar or a sugar substitute, which is a medicine having an unpleasant taste, for example, a bitter taste, and is made into a liquid form for easy consumption. In particular, it is a formulation suitable for consumption by infants and children. The syrup may contain, in addition to purified water and an extract, an antibacterial preservative, a flavoring agent, a coloring agent, or a sweetener used to provide sweetness and viscosity, but is not limited thereto. Examples of sweeteners that may be included in the syrup include, but are not limited to, sucrose, mannitol, sorbitol, xylitol, aspartame, stevioside, fructose, lactose, sucralose, saccharin, or menthol.

[0072] In tablet formulations, the active ingredient drug may be present in an amount of from about 0.05% to about 95% by weight of the total weight of the tablet, more typically from about 2% to about 50% by weight of the formulation. The tablet may also contain a disintegrant, which comprises from about 0.5% to about 35% by weight, more typically from about 2% to about 25% by weight of the formulation. Examples of disintegrants include, but are not limited to, lactose, starch, sodium starch glycolate, crospovidone, croscarmellose sodium, maltodextrin, or mixtures thereof.

[0073] Suitable lubricants included for manufacturing the tablets may be present in an amount of from about 0.1% to about 5% by weight, and examples of lubricants include, but are not limited to, talc, silicon dioxide, stearic acid, calcium, zinc or magnesium stearate, sodium stearyl fumarate, and the like.

[0074] Gelatin, polyethylene glycol, sugar, gum, starch, polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, etc. can be used as binders for manufacturing into tablets, and mannitol, xylitol, lactose, dextrose, sucrose, sorbitol, starch, microcrystalline cellulose, etc. can be used as suitable diluents for manufacturing into tablets, but are not limited thereto.

[0075] Solubilizing agents that may optionally be included in the tablet may be used in an amount of about 0.1 wt% to about 3 wt% based on the total weight of the tablet, and examples thereof include polysorbates, sodium lauryl sulfate, sodium dodecyl sulfate, propylene carbonate, diethylene glycol monoethyl ether, dimethyl isosorbide, polyoxyethylene glycolated natural or hydrogenated castor oil, HCOR. TM(Nikkol), oleic acid ester, gelucire TM ), caprylic / caprylic acid mono / diglycerides, sorbitan fatty acid esters, solutol HS TM The following may be used, but are not limited to:

[0076] From another aspect, the present invention relates to a health functional food composition for preventing or improving neurological, mental and cognitive disorders, comprising the above L-serine hydrochloride as an active ingredient.

[0077] In the present invention, "health functional food" means a food group that has added value by using physical, biochemical, bioengineering techniques to make the function of the food work and express it for a specific purpose, or a food that has been designed and processed so that the body's internal regulatory function regarding biological defense rhythm regulation, disease prevention, and recovery, etc. of the food composition, is sufficiently expressed in the body. For the purpose of the present invention, the health functional food may mean a health functional food for improving neurological, mental, and cognitive disorders, for example, a health functional food for preventing and improving memory loss.

[0078] The above health functional food composition can be manufactured using methods commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during manufacturing. Furthermore, the formulation of the above health functional food composition can be manufactured without limitation, as long as it is a formulation recognized as a food composition in the art.

[0079] Foods according to the present invention include, for example, various foods, beverages, gums, tea, vitamin complexes, functional foods, etc. Foods include, but are not limited to, special nutritional foods (e.g., formulated milk, infant and baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g., ramen, noodles, etc.), bread, health supplements, seasoned foods (e.g., soy sauce, soybean paste, red pepper paste, mixed paste, etc.), sauces, confectionery (e.g., snacks), candies, chocolates, gums, ice cream, dairy products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (e.g., various kimchi, pickled vegetables, etc.), beverages (e.g., fruit drinks, vegetable drinks, soy milk, fermented drinks, etc.), natural seasonings (e.g., ramen soup, etc.), food additives, etc. The foods, beverages, or food additives may be manufactured by a conventional manufacturing method.

[0080] When the health functional food composition of the present invention is used as a health functional food additive, the composition can be added as is or used together with other health functional food ingredients, and can be used appropriately according to a conventional method. The mixing amount of the active ingredient can be appropriately determined depending on the intended use. Generally, when manufacturing a food or beverage, the composition of the present invention can be added in an amount of preferably about 50 parts by weight or less, more preferably 25 parts by weight or less, relative to the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range. In addition, it can be used in an amount greater than the above range as long as there is no problem in terms of safety.

[0081] The health functional food composition of the present invention may contain, in addition to containing the hydrochloride of L-serine as an effective ingredient, conventional food composition additives such as natural carbohydrates or flavoring agents as additional ingredients. Examples of the natural carbohydrates may include conventional sugars such as monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. Examples of the flavoring agents may include natural flavoring agents (e.g., thaumatin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (e.g., saccharin, aspartame, etc.).

[0082] In addition, the health functional food composition may contain, in addition to the hydrochloride of L-serine, which is an effective ingredient, nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the food composition of the present invention may contain fruit pulp for producing natural fruit juice, fruit juice drinks, and vegetable drinks.

[0083] Any description that overlaps with the pharmaceutical composition of the present invention described above is omitted.

[0084] In another aspect, the present invention relates to a method for preventing or treating a neurological, mental and cognitive disorder disease in a subject, comprising administering to the subject the L-serine hydrochloride.

[0085] In the present invention, the term "subject" is used synonymously with an individual, and the subject may be a mammal, for example, a human, a rat, a cow, a horse, a pig, a dog, a sheep, a goat, or a cat.

[0086] Any description that overlaps with the pharmaceutical composition of the present invention described above is omitted.

[0087] In another aspect, the present invention relates to the use of L-serine hydrochloride for preventing or treating neurological, mental and cognitive disorders.

[0088] Any description that overlaps with the pharmaceutical composition of the present invention described above is omitted.

[0089] In another aspect, the present invention relates to the use of the L-serine hydrochloride for preparing a medicament for preventing or treating neurological, mental and cognitive disorders.

[0090] Any description that overlaps with the pharmaceutical composition of the present invention described above is omitted.

[0091] All matters mentioned in the compositions, treatment methods and uses of the present invention apply equally unless they are contradictory.

[0092] Hereinafter, the present invention will be described in detail, using examples and the like, to aid understanding. However, the examples according to the present invention may be modified in various different forms, and the scope of the present invention should not be construed as being limited to the following examples. The examples of the present invention are provided to more fully explain the present invention to those of average skill in the art.

[0093] Example

[0094] Example 1. Preparation of L-serine hydrochloride

[0095] 4.0 kg of L-Serine and 20 L of IPA (Isopropyl alcohol) were placed in a reactor at room temperature and stirred for 10 minutes. Then, 12.6 kg of 3M HCl in IPA was slowly added dropwise while maintaining room temperature. The mixture was stirred at room temperature for 12 hours, filtered, and washed with 8 L of IPA. Subsequently, vacuum drying was performed at 50°C to obtain a white solid with a yield of 93.5%.

[0096] The obtained solid was dissolved in D2O and 1H-NMR was observed, and it was confirmed that there were three hydrogens in the characteristic peaks at 3.8 to 4.2.

[0097]

[0098] Example 2. Preparation of L-serine hydrochloride

[0099] 4.0 kg of L-Serine and 25 L of IPA were placed in a reactor at room temperature and stirred for 10 minutes. 9.1 kg of 4 M HCl in IPA was slowly added dropwise while maintaining room temperature. The mixture was stirred at room temperature for 12 hours, filtered, and washed with 8 L of IPA. Subsequently, vacuum drying was performed at 50°C to obtain a white solid with a yield of 92.5%.

[0100] The obtained solid was dissolved in D2O and H-NMR was observed, and it was confirmed that there were three hydrogens in the characteristic peaks at 3.8 to 4.2.

[0101]

[0102] Example 3. Preparation of L-serine hydrochloride

[0103] 4.0 kg of L-Serine and 25 L of IPA were placed in a reactor at room temperature and stirred for 10 minutes. 9.1 kg of 4 M HCl in IPA was slowly added dropwise while maintaining room temperature. The mixture was stirred for 12 hours at 10°C or lower, filtered, and washed with 8 L of IPA. Subsequently, vacuum-dried at 50°C, a white solid was obtained with a yield of 93.7%.

[0104] The obtained solid was dissolved in D2O and H-NMR was observed, and it was confirmed that there were three hydrogens in the characteristic peaks at 3.8 to 4.2.

[0105]

[0106] Example 4. Preparation of L-serine hydrochloride

[0107] 4.0 kg of L-Serine and 25 L of ethanol were placed in a reactor at room temperature and stirred for 10 minutes. 9.0 kg of 4 M HCl in ethanol was slowly added dropwise while maintaining room temperature. The mixture was stirred for 12 hours at 10°C or lower, filtered, and washed with 8 L of IPA. Subsequently, vacuum drying was performed at 50°C to obtain a white solid with a yield of 91.5%.

[0108] The obtained solid was dissolved in D2O and H-NMR was observed, and it was confirmed that there were three hydrogens in the characteristic peaks at 3.8 to 4.2.

[0109]

[0110] Example 5. Preparation of L-serine hydrochloride

[0111] 4.0 kg of L-Serine and 25 L of ethanol were placed in a reactor at room temperature and stirred for 10 minutes. 9.0 kg of 4 M HCl in ethanol was slowly added dropwise while maintaining room temperature. The mixture was stirred for 12 hours at 10°C or lower, filtered, and washed with 8 L of acetone. Subsequently, vacuum drying was performed at 50°C to obtain a white solid with a yield of 90.9%.

[0112] The obtained solid was dissolved in D2O and H-NMR was observed, and it was confirmed that there were three hydrogens in the characteristic peaks at 3.8 to 4.2.

[0113]

[0114] Example 6-1. Differential Scanning Calorimeter (DSC) Analysis

[0115] The L-serine hydrochloride prepared in Example 3 was analyzed by differential scanning calorimetry (DSC), confirming the formation of crystalline L-serine hydrochloride (Fig. 1). The formed crystalline L-serine hydrochloride is characterized by the peaks appearing in the DSC thermogram. The crystalline L-serine hydrochloride according to the present invention exhibited one endothermic peak at approximately 150°C (Fig. 1).

[0116]

[0117] Example 6-2. Powder X-ray diffraction (PXRD) analysis

[0118] Powder X-ray powder rotation patterns are widely used to distinguish between crystalline and hydrated forms of drugs due to their unique properties. Therefore, the powder X-ray pattern of L-serine hydrochloride was measured.

[0119] The L-serine hydrochloride prepared in Example 3 was analyzed by powder X-ray diffraction (PXRD), and it was confirmed that crystalline L-serine hydrochloride was formed (Fig. 2). The formed crystalline L-serine hydrochloride is characterized by a characteristic 2θ (2theta) diffraction angle peak appearing in PXRD. Specifically, the crystalline L-serine hydrochloride according to the present invention exhibits characteristic peaks at diffraction angles (2θ±0.2) of 16.4°, 16.9°, 18.8°, 19.9°, 20.7°, 22.5°, 25.3°, 26.6°, 27.2°, 27.8°, 29.0°, 30.8°, 32.4°, 33.1°, 37.5°, and 38.1°, respectively, when measured in a PXRD analysis at a scanning range (2θ): 3 to 50° and a scan speed: 6 deg / min (Fig. 2).

[0120]

[0121] Example 6-3. Comparison of Differential Scanning Calorimetry (DSC) of L-serine hydrochloride and L-serine

[0122] As a result of analyzing L-serine hydrochloride and L-serine manufactured in Example 3 using Differential Scanning Calorimeter (DSC), L-serine hydrochloride showed an endothermic peak at about 150°C as confirmed in Example 6-1, whereas L-serine showed an endothermic peak at about 226°C (Fig. 3).

[0123]

[0124] Example 6-4. Comparison of powder X-ray diffraction (PXRD) analysis of L-serine hydrochloride and L-serine

[0125] The powder X-ray diffraction (PXRD) angle values ​​of L-serine have characteristic peaks at diffraction angles (2θ±0.2) of 18.4°, 19.0°, 18.8°, 19.9°, 20.7°, 22.8°, 24.8°, 26.9°, 28.2°, 30.5°, 31.8°, 33.3°, 34.9°, 37.4°, and 39.5°, respectively (Fig. 4).

[0126] The numerical values ​​of the powder X-ray diffraction (PXRD) angles of L-serine hydrochloride were different from those confirmed in Example 6-2, confirming that it was a new crystalline form.

[0127]

[0128] Example 6-5. Chloride titration of L-serine hydrochloride

[0129] Approximately 0.2 g of L-serine hydrochloride prepared in Example 3 was dissolved in water and titrated with a 0.1 mol / L silver nitrate solution, and the result showed that it contained approximately 22.5% chloride (Fig. 5).

[0130]

[0131] Example 6-6. Comparison of aqueous solubility of L-serine hydrochloride and L-serine.

[0132] The water solubility of L-serine hydrochloride and L-serine prepared in Example 3 was compared. While the water solubility of L-serine was 300 mg / ml, the water solubility of L-serine hydrochloride was 1560 mg / ml, confirming that the water solubility of L-serine hydrochloride was more than 5 times that of L-serine. In addition, no discoloration or precipitation occurred even when stored at room temperature for one month in an aqueous solution state.

[0133]

[0134] Below, a mouse PK experiment was conducted to confirm the improvement in the pharmacokinetic profile of L-serine hydrochloride prepared in Example 3.

[0135]

[0136] Experimental Example 1

[0137] Before drug administration -6, -4, -2, 0 hours and after drug administration 0.25, 0.5, 1, 3, 6, 9, 12 hours, the experimental mice were placed in a rigid frame, a 2 mm incision was made at the tip of the tail, and approximately 40-50 μl of blood was collected into a heparin capillary tube while massaging the tail. After blood collection, the mice were euthanized in a CO2 chamber and the carcasses were disposed of by cervical dislocation. The mice were transferred to 2 ml tubes and centrifuged at 2000 × g for 10 minutes to separate the serum, which was then transferred to new tubes at approximately 20-25 μl per sample. For LC-MS / MS analysis, 10 μl of serum was transferred to new tubes, and the remaining serum was stored in a deep freezer at -80 °C. The L-serine hydrochloride prepared in Example 3 was diluted in water to a concentration of 10 mg / mL and the internal standard D, L-Serine-2,3,3-d3, was diluted in water to a concentration of 100 μg / mL. The L-serine hydrochloride solution was diluted in a solvent with a ratio of acetonitrile / water / trifluoroacetic acid (v / v / v, 96 / 5 / 0.5) to a concentration of 0.01 to 41.38 μg / mL to prepare a standard reagent necessary for preparing a calibration curve. The internal standard D, L-serine-2,3,3-d3 stock solution was prepared at a concentration of 60 μg / mL for preparing a calibration curve and at 2 μg / mL for spiking into samples. To prepare the reagents required for the calibration curve, 10 μL of the diluted stock solution and 10 μL of the internal standard solution (DL-serine-2,3,3-d3, 60 μg / mL) were spiked into 980 μL of a solvent with a ratio of acetonitrile / water / trifluoroacetic acid (v / v / v, 96 / 5 / 0.5). 10 μL of the rat plasma to be analyzed and the internal standard solution (D, L-serine-2,3,3-d3, 3.6 μg / mL) was spiked into 10 μL and 980 μL of the initial condition solvent acetonitrile / water / trifluoroacetic acid (v / v / v, 96 / 5 / 0.5) was added, and vortexed for 1 minute. The mixed solution was centrifuged at 16,000 g for 10 minutes, and the supernatant was separated and injected into LC-MS / MS with 5 μL. The ultra-high-performance liquid chromatography system consisted of a pump (LC-40D X3), autosampler (SIL-40CX3), column oven (CTO-40C), and system controller (SCL-40) from Shimadzu (Duisburg, Germany), and the mass spectrometer used was QTRQP LC-MS / MS 6500+ from SCIEX (Framingham, MA, USA). The column used was a CROWNPAK CR-I (+) [3.0 x 150 mm, 5 micron] from DAICEL (Osaka, Tokyo), and the column oven was set to 30°C. The mobile phase was acetonitrile / water / trifluoroacetic acid (v / v / v, 96 / 5 / 0.5), and the elution was performed under isocratic conditions. Mass spectrometry detection was performed under MRM (multiple reaction monitoring) conditions in ESI+ mode. MS / MS parameters were as follows: ion spray voltage, 5500 V; temperature 500°C; curtain gas, 30 psi; ion source gas (GS1), 50 psi, ion source gas (GS2), 50 psi.Mass spectrometry was performed by optimizing the declustering potential (DP), entrance potential (EP), collision energy (CE), and collision cell exit potential (CXP) values ​​under MRM conditions in manual tuning mode using a syringe infusion pump. Data acquisition and processing were performed using AB SCIEX Analyst software.

[0138] As a result, as shown in Table 1, it was confirmed that L-serine hydrochloride had a significantly increased half-life and blood drug concentration compared to L-serine (Figure 6).

[0139] Parameter L-serine L-serine hydrochloride (Example 3)t 1 / 2 (h)1.21±0.481.80±0.27T max (h)0.250.62C max (μg / ml)67.50±25.9277.69±8.49AUC 0-6 (μg / ml*h)81.46±22.54141.34±37.40Cl / F((mg / kg) / (μg / ml) / h)4.92±1.203.77±0.94

[0140]

[0141] Experimental Example 2

[0142] In order to conduct a behavioral experiment on VPA (valproic acid)-induced autism model mice using L-serine hydrochloride prepared in Example 3, 600 mg / kg of VPA dissolved in 0.9% NaCl saline was subcutaneously administered to pregnant dams to obtain 14 VPA-induced autism model mice. Meanwhile, 8 pups from dams that were not administered VPA were used as a control group. For 14 days after birth, (i) the vehicle was orally administered to the control mice (n=8) (Ctrl + Vehicle treatment group), (ii) the vehicle was orally administered to some of the VPA-induced autism model mice (n=8) (VPA + Vehicle treatment group), and (iii) the L-serine hydrochloride prepared in Example 3 was orally administered to another some of the VPA-induced autism model mice (n=6) (VPA + L-serine hydrochloride treatment group). The oral administration method administered the drug at a dose of 10 ml / kg per day based on the individual's body weight, which corresponds to the same molar amount as administering 400 mg / kg of L-serine hydrochloride to humans.

[0143] Thirty-five days after birth, the mice's social interaction abilities were tested using a three-chamber test. Social novelty was assessed by assessing the interaction time between the first encounter with a novel mouse and the re-encounter with a familiar mouse. Sociability assessed the degree of social interaction between mice and other mice, and was measured by assessing the interaction time of the mice toward other mice on one side of an empty box.

[0144] As a result, vehicle-treated VPA mice (VPA) showed significantly lower social novelty and sociability compared to vehicle-treated control mice (Control). In contrast, L-serine hydrochloride-treated VPA mice (VPA + L-serine hydrochloride) showed statistically significant improvements in both social novelty and sociability assessments, completely restoring social novelty and sociability to levels above the control group (Figures 8A and 8B).

Claims

1. (a) A step of mixing L-serine of the following chemical formula 1 with an organic solvent; (b) a step of adjusting the pH of the mixed solution of step (a) with hydrochloric acid diluted in an organic solvent; (c) a step of forming a salt by stirring the solution whose pH has been adjusted in step (b); and (d) A step of filtering the salt produced in step (c) and then washing the excess hydrochloric acid with an organic solvent. Method for producing L-serine hydrochloride comprising [Chemical Formula 1] .

2. A method for producing L-serine hydrochloride in the first paragraph, wherein the organic solvent in step (a) is selected from the group consisting of acetone, methanol, ethanol, isopropanol, butanol, hexanol, ethyl acetate, toluene, methyl-t-butyl ether, and methylene chloride.

3. A method for producing L-serine hydrochloride, wherein in step (b) of paragraph 1, the pH is adjusted to 1 to 5.

4. A method for producing L-serine hydrochloride in the first paragraph, wherein the organic solvent in step (b) is selected from the group consisting of acetone, methanol, ethanol, isopropanol, butanol, hexanol, ethyl acetate, toluene, methyl-t-butyl ether, and methylene chloride.

5. A method for producing L-serine hydrochloride in the first paragraph, wherein the stirring time in step (c) is 2 to 48 hours.

6. A method for producing L-serine hydrochloride, wherein in step (c), stirring is performed at a temperature below room temperature.

7. A method for producing L-serine hydrochloride in the first paragraph, wherein the organic solvent of step (d) is selected from the group consisting of acetone, methanol, ethanol, isopropanol, butanol, hexanol, ethyl acetate, toluene, methyl-t-butyl ether, methylene chloride, cyclohexane, and pentane.

8. A method for producing L-serine hydrochloride in the first paragraph, wherein the organic solvent of step (d) has a moisture content of 1% or less.

9. A method for producing L-serine hydrochloride, wherein the method further comprises a step of (e) drying to remove the solvent after step (d) in paragraph 1.

10. A method for producing L-serine hydrochloride, wherein in step (e) of claim 9, the drying temperature is 20 to 80°C.

11. L-serine hydrochloride of the following chemical formula 2 manufactured by the manufacturing method of Article 1 [Chemical formula 2] .

12. A pharmaceutical composition for the prevention or treatment of neurological, mental and cognitive disorders, comprising L-serine hydrochloride of Article 11 as an active ingredient.

13. A pharmaceutical composition according to claim 12, wherein the neurological, mental and cognitive disorder is selected from the group consisting of autism spectrum, bipolar disorder, Tourette's, tics, Prader-Willi, attention deficit hyperactivity disorder (ADHD), Fragile-X syndrome, dementia, Alzheimer's, Parkinson's, schizophrenia, depression and memory impairment.

14. A pharmaceutical composition in any one dosage form selected from the group consisting of tablets, lozenges, capsules, granules, powders, and liquids in the 12th paragraph.

15. A pharmaceutical composition in claim 14, wherein L-serine hydrochloride in the purified formulation is in an amount of 0.05 wt% to 95 wt% based on the total weight of the tablet.

16. A health functional food composition for preventing or improving neurological, mental and cognitive disorders, containing L-serine hydrochloride of Article 11 as an effective ingredient.

17. In claim 16, the health functional food composition is selected from the group consisting of autism spectrum, bipolar disorder, Tourette's, tics, Prader-Willi, attention deficit hyperactivity disorder (ADHD), Fragile-X syndrome, dementia, Alzheimer's, Parkinson's, schizophrenia, depression, and memory impairment.

18. A method for preventing or treating a neurological, mental and cognitive disorder disease in a subject, comprising the step of administering L-serine hydrochloride of clause 11 to the subject.

19. Use of L-serine hydrochloride of Article 11 for preventing or treating neurological, mental and cognitive disorders.

20. Use of L-serine hydrochloride of claim 11 for the manufacture of a medicament for preventing or treating neurological, mental and cognitive disorders.

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