Method for producing glycolipid carboxylic acid

Abstract

The present invention relates to a process for the preparation of glycolipid carboxylic acids of formula I by selective enzymatic hydrolysis of the corresponding esters, said process providing for the preparation of compounds of formula I in high yields by an environmentally friendly, robust, economical, simple and convenient method. JPEG2024522887000081.jpg2851

Description

[Technical field]

[0001] CROSS REFERENCE TO RELATED APPLICATIONS: This application claims priority to Indian Provisional Patent Application No. 202141028550, filed on June 25, 2021, the entire contents of which are specifically incorporated herein by reference.

[0002] The present invention relates to synthetic organic chemistry. The present invention relates to a method for producing glycolipid carboxylic acids and derivatives thereof. The method comprises the selective enzymatic hydrolysis of glycolipid carboxylic acid esters. In particular, the method comprises the production of compounds of formula I by the selective enzymatic hydrolysis of glycolipid carboxylic acid esters to the corresponding carboxylic acids. The method provides an economical and environmentally friendly approach for the production of complex organic compounds in high yields. [Background technology]

[0003] Glycolipids are essential components of cell membranes, consisting of sugar and lipid moieties. Glycolipids are widely known to be important for cell aggregation, dissociation, and also for specific cell contact and signal transduction. It also acts as a promising delivery system for RNA interference (RNAi) therapeutics. This application has been widely used and investigated as a possible therapeutic agent for reversibly silencing any gene. To achieve the clinical potential of RNAi, a delivery material is needed to transport short interfering RNA (siRNA) to the active site in the cells of the target tissue.

[0004] Glycolipids such as N-acetylgalactosamine (GalNAc) and its derivatives are prominent among the compounds for drug delivery. The conjugates have been developed by directly conjugating the delivery agent to the siRNA cargo. This approach leads to a well-defined single-component system that uses only equimolar amounts of delivery material and siRNA. Therapeutic agents require that the compound is in its native form to minimize undesirable side effects and dosage. However, obtaining the native compound is difficult, especially for the complex compounds.

[0005] N-acetylgalactosamine (GalNAc) or its derivatives are sensitive glycolipids due to their large structure and easily accessible functional groups.

[0006] Migawa, Michael T. et al., "A convenient synthesis of 5'-triantennary N-acetyl-galactosamine clusters based on nitromethanetrispropionic acid" Bioorganic & Medicinal Chemistry Letters 26.9 (2016): 2194-2197, reports the development of a method for the synthesis of several triantennary GalNAc clusters based on a nitromethanetrispropionic acid core. The synthetic approach was used in a one-pot, seven-reaction procedure to obtain a variety of triantennary GalNAc conjugates. ASO The present invention also includes pentafluorophenolic ester intermediates which can be used to prepare the

[0007] WO 2014 / 025805A1 reports the preparation of a three-branched GalNAc derivative.

[0008] However, the prior art methods for preparing said compounds involve harsh reaction conditions, multiple steps, and low yields of the desired compounds. The conventional preparation methods are difficult and uneconomical for large-scale production.

[0009] Considering the importance of glycolipid compounds, it is necessary to develop a stable, economical and environmentally friendly process for the preparation of said complex glycolipids and their potential intermediates. Surprisingly, the present inventors have developed an efficient process for the preparation of glycolipid carboxylic acids and their derivatives, which meets the above requirements and improves upon the shortcomings of the prior art.

[0010] Objective of the invention It is an object of the present invention to provide an efficient method for producing glycolipid carboxylic acids and derivatives thereof.

[0011] It is another object of the present invention to provide an efficient process which is robust, economical, simple, less step and environmentally friendly.

[0012] It is another object of the present invention to provide an efficient process which results in glycolipid carboxylic acids and their derivatives in high yield and purity.

[0013] It is yet another object of the present invention to provide methods for preparing compounds of formula I. Summary of the Invention

[0014] According to one embodiment of the present invention, there is provided a method for producing glycolipid carboxylic acids and derivatives thereof. The glycolipid carboxylic acids and derivatives produced are N-acetylgalactosamine (GalNAc) derivatives.

[0015] Another aspect of the present invention provides a process for the preparation of compounds of formula I and intermediates thereof, comprising selective enzymatic hydrolysis. Compounds of formula I may be used as key intermediates in the preparation of therapeutically important glycolipids.

[0016] According to one embodiment of the present invention, a compound of formula (I): [ka] A method for preparing the compound of formula (I) is provided.

[0017] According to another aspect of the present invention, a compound of formula (I'): [ka] A method for preparing the compound of formula (I) is provided.

[0018] According to yet another aspect of the present invention, a compound represented by the formula (1-e): [ka] A method for preparing the compound of formula (I) is provided.

[0019] According to yet another aspect of the present invention there are provided compounds of formula (I) and formula (I').

[0020] Detailed Description of the Invention The following description, which refers to the accompanying drawings, is provided to aid in a comprehensive understanding of exemplary embodiments of the present invention, and although it includes various specific details to aid in understanding, it should be considered as merely illustrative.

[0021] Therefore, those skilled in the art will recognize that changes and modifications of the embodiments described herein can be made without departing from the scope of the present invention. Moreover, descriptions of known functions and constructions are omitted for clarity and conciseness.

[0022] The terms and words used in the following description and claims are not limited to their bibliographical meanings, but are used by the inventors simply to enable a clear and consistent understanding of the present invention. Therefore, it should be clear to those skilled in the art that the following description of exemplary embodiments of the present invention is provided for illustrative purposes only, and is not intended to limit the scope of the present invention as defined by the appended claims and their equivalents.

[0023] The singular forms "a," "an," and "the" should be understood to include plural referents unless the context clearly indicates otherwise.

[0024] Features described and / or illustrated with respect to one embodiment may be used in the same or similar manner in one or more other embodiments and / or in combination with or instead of features of the other embodiments.

[0025] It should be emphasized that as used in this specification, "comprises" is understood to specify the presence of stated features, steps or components, but does not exclude the presence or addition of one or more other features, steps or components or combinations (groups) thereof.

[0026] The present invention relates to a method for the selective enzymatic hydrolysis of glycolipid carboxylic acid esters. In particular, the method involves the production of glycolipid carboxylic acids and their derivatives by selective enzymatic hydrolysis of the corresponding glycolipid carboxylic acid esters. The method provides an economical and environmentally friendly approach for the production of complex organic compounds in high yields.

[0027] The present invention provides an efficient method for preparing compounds of formula I by enzymatic hydrolysis of the ester to the acid and enzymatic treatment. [ka]

[0028] The present invention relates to a compound of formula (I): [ka] [During the ceremony, "n" ranges from 0 to 15; A1, A2 and A3 each independently represent H or [ka] Selected from; However, A1, A2 and A3 cannot all be H; G is the following group: [ka] Selected from; X is the following group [ka] Selected from; Y is [ka] Selected from; Z is the following group [ka] Selected from; where R2 is selected from alkyl, aryl, alkoxy, aryloxy, or arylalkyl groups; "l", "m" and "p" each independently range from 0 to 15. A method for producing a compound of the formula (a) in the presence of an enzyme and water, [ka] wherein R is selected from an alkyl, aryl or aralkyl group. hydrolyze the compound; (b) stirring the reaction mixture of step (a), concentrating, drying and diluting with a solvent; (c) removing the solid precipitate, which is the enzyme, from the reaction mixture of step (b) and purifying it to obtain a compound of formula (I); A method is provided, comprising the steps of:

[0029] In certain embodiments, A1, A2 and A3 are each independently [ka] and the substituent [ka] The following groups: [Table 1] [During the ceremony, R2 is selected from alkyl, aryl, alkoxy, aryloxy, or arylalkyl groups; "l", "m", "n" and "p" and "q" each independently range from 0 to 15. is selected from.

[0030] In certain embodiments, the enzyme in step (a) is selected from lipase, protease, amylase, trypsin, papain, or a combination thereof, hi certain embodiments, the enzyme is lipase.

[0031] In one embodiment, the enzyme in step (a) is present in an amount in the range of 1 to 100% by weight.

[0032] In one embodiment, the enzymatic hydrolysis in step (a) is carried out at 20-45° C. for 1-48 hours.

[0033] In certain embodiments, the solvent in step (b) is selected from water, methanol, ethanol, isopropyl alcohol, methyl tert-butyl ether, dichloromethane, ethyl acetate, acetone, dimethylformamide, tetrahydrofuran, and acetonitrile, or a combination thereof.

[0034] In certain embodiments, the groups are straight chain, branched chain, cyclic groups with or without functional groups selected from, but not limited to, halogen, nitro, amine, aldehyde, carbonyl, hydroxyl, and the like.

[0035] In certain embodiments, alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, and the like.

[0036] In certain embodiments, the aryl group is a phenyl, naphthyl group, and the like.

[0037] In certain embodiments, the aralkyl or substituted aralkyl is benzyl, p-methoxybenzyl, and the like.

[0038] Preferably, the present invention provides a compound of formula (I') by enzymatic hydrolysis: [ka] (wherein G, X, Y, Z and R2 are as defined above). The present invention provides a method for preparing the compound of formula (I). [ka]

[0039] A detailed process is shown in Scheme C below: [ka]

[0040] In another embodiment, instead of the glycolipid carboxylic acid, a compound of the formula: [ka] The glycolipid amine is used as a starting reactant and undergoes the same treatment steps as in step 2 and step 3 to give the glycolipid carboxylic acid (compound of formula I').

[0041] The method includes the steps of: (i) Formula (zb): [ka] or glycolipid carboxylic acid of the formula (z-bb): [ka] A reactant selected from glycolipid amine compounds of formula (zc): [ka] to an ester of formula (Ia)': [ka] forming a compound of; (ii) The ester compound of formula (Ia)' is hydrolyzed in the presence of an enzyme and water to give a compound of formula (I)'.

[0042] In certain embodiments, reaction parameters such as coupling agent, solvent, temperature, pH, reaction time, etc. vary depending on the enzyme and the compound of Formula I or a derivative thereof of interest.

[0043] In another embodiment, the pH of the reaction is maintained between 2 and 9; the reaction temperature can vary between 0° C. and 150° C. The reaction time varies between 2 and 72 hours or until the reaction is complete.

[0044] In another embodiment, the solvent for the reaction is selected from protic or aprotic solvents or a combination thereof in an appropriate ratio; wherein said protic are water, methanol, ethanol, isopropyl alcohol and said aprotic solvents are methyl tert-butyl ether, dichloromethane, ethyl acetate, acetone, dimethylformamide, tetrahydrofuran and acetonitrile.

[0045] In certain embodiments, the coupling in step (i) is carried out in the presence of a coupling agent selected from the group comprising hydroxybenzotriazole (HOBt), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), ethyl-(N',N'-dimethylamino)propylcarbodiimide hydrochloride (EDC·HCL), or a combination thereof.

[0046] In certain embodiments, the coupling reaction of step (i) is carried out in the presence of a base selected from N,N-diisopropylethylamine (DIPEA).

[0047] In one embodiment, the glycolipid carboxylic acid of formula (zb) in step (i) is converted to a glycolipid carboxylic acid of formula (za) in the presence of an enzyme and water: [ka] to obtain a glycolipid carboxylic acid of formula (zb). EXAMPLES

[0048] The following examples are meant to illustrate the present invention, and are presented to illustrate the invention and should not be considered as limiting the scope of the invention.

[0049] Example A: experiment: The detailed process is shown in Scheme C. General method A (step 1): The enzyme (20 wt%) was added to a solution of glycolipid carboxylate (1 equiv.) in water (5 V) at 20-25°C. The reaction mixture was stirred at 20-25°C for 24-72 h. Completion of the reaction was monitored by TLC. The reaction mixture was concentrated to dryness under reduced pressure at 35°C. The resulting residue was diluted with DCM (15 V) and stirred for 20 min. The solid precipitate (enzyme) was removed by filtration and the filtrate was concentrated under reduced pressure at 35°C to give the glycolipid carboxylate.

[0050] General method B (step 2): To a solution of glycolipid carboxylic acid (4.5 equiv.) in DCM (10V) at 0-5°C, HOBt (0.1 equiv.), DIPEA (10 equiv.) and EDC·HCl (4.5 equiv.) were added. The reaction mixture was stirred at the same temperature for 10-15 min, after which the amine (1.0 equiv.) dissolved in DMF:DCM (1:5) was added dropwise. The resulting reaction mixture was stirred at 20-35°C for 16 h. The reaction was concentrated to dryness, slurried with MTBE (10V×3) and decanted. The residue was dissolved in DCM (10V) and washed with water (5V×2), and the organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain the compound of formula (Ia)'.

[0051] General method C ((Step 3): The enzyme (5 wt%) was added to a solution of glycolipid carboxylate (1 equiv.) in water (5 V) at 20-25°C. The reaction mixture was stirred at 20-25°C for 12-24 h. Completion of the reaction was monitored by TLC. The reaction mixture was concentrated to dryness under reduced pressure at 35°C. The resulting residue was diluted with DCM (15 V) and stirred for 20 min. The solid precipitate (enzyme) was removed by filtration and the filtrate was concentrated under reduced pressure at 35°C to give glycolipid carboxylate (compound of formula I').

[0052] In another embodiment, the formula [ka] The glycolipid amine of formula I' is used as a starting reactant instead of the glycolipid carboxylic acid, and the glycolipid carboxylic acid (compound of formula I') is obtained through the same process steps as in General Method B and General Method C.

[0053] A variety of commercially available enzymes are utilized for the hydrolysis as shown in Table 1. In yet another embodiment, the amount of enzyme for the reaction can vary between 1-100% wt / wt compared to the reactants. [Table 2-1] [Table 2-2]

[0054] Example 1: [ka] Step 1: Preparation of 5-{[4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentanoic acid (1-b) [ka] Starting from 1-a, following general procedure A, compound 1-b was obtained as a white solid (50 g, 89%).

[0055] Step 2: Preparation of methyl 11-{[1,3-bis(2-{[3-(5-{[4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)-2-[(2-{[3-(5-{[4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)methyl]propan-2-yl]carbamoyl}undecanoate (1-d) [ka] Starting from 1-b, following general method B, compound 1-d was obtained as a white solid (1.5 g, 80%).

[0056] Step 3: Preparation of 11-{[1,3-bis(2-{[3-(5-{[4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)-2-[(2-{[3-(5-{[4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)methyl]propan-2-yl]carbamoyl}undecanoic acid (1-e). [ka] Starting from 1-d, following general method B, compound 1-e was obtained as an off-white foamy solid (25 g, 98%).

[0057] Example 2: [ka] Step 1: Preparation of 3-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethoxy]propanoic acid (2-b) [ka] Starting from 2-a, following general procedure A, compound 2-b was obtained as a white solid (1.0 g, 86%).

[0058] Step 2: Preparation of methyl 11-{[1,3-bis({2-[(3-{3-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethoxy]propanamido}propyl)carbamoyl]ethoxy})-2-({2-[(3-{3-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethoxy]propanamido}propyl)carbamoyl]ethoxy}methyl)propan-2-yl]carbamoyl}undecanoate (2-d) [ka] Starting from 2-a, following general method B, compound 2-d was obtained as a white solid (1.7 g, 85%). Under reduced pressure, 1.7 g (85%) of compound (2-d) was obtained as an off-white foam.

[0059] Step 3: Preparation of 11-{[1,3-bis({2-[(3-{3-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethoxy]propanamido}propyl)carbamoyl]ethoxy})-2-({2-[(3-{3-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethoxy]propanamido}propyl)carbamoyl]ethoxy}methyl)propan-2-yl]carbamoyl}undecanoic acid (2-e). [ka] Starting from 2-d, following general procedure C, compound 2-e was obtained as an off-white foamy solid (500 mg, 97%).

[0060] Example 3: [ka] Step 1: Preparation of 3-{2-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethoxy]ethoxy}propanoic acid (3-b) [ka] Starting from 3-a, following general procedure A, compound 3-b was obtained as a white solid (2.0 g, 88%).

[0061] Step 2: Methyl 4-{[1-(2-{[3-(3-{2-[2-(2-{[(2R,5R)-5-(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamido-4-(formyloxy)oxan-2-yl]oxy}ethoxy)ethoxy]ethoxy}propanamido)propyl]carbamoyl}ethoxy)-3-(2-{[3-(3-{2-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamido Preparation of (3-d) [ka] Starting from 3-b and following general method B, compound 3-d was obtained as an off-white foam (4.0 g, 87%).

[0062] Step 3: 4-{[1-(2-{[3-(3-{2-[2-(2-{[(2R,5R)-5-(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamido-4-(formyloxy)oxan-2-yl]oxy}ethoxy)ethoxy]ethoxy}propanamido)propyl]carbamoyl}ethoxy)-3-(2-{[3-(3-{2-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamido Preparation of (3-(2-[(3-{2-[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethoxy]ethoxy}propanoyl)carbamoyl]ethoxy}methyl)propan-2-yl]carbamoyl}butanoic acid (3-e). [ka] Starting from 3-d, following general procedure C, compound 3-e was obtained as an off-white foamy solid (1.5 g, 96%).

[0063] Example 4: [ka] Step 1: Preparation of 12-{[1,3-bis(2-{[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl}ethoxy)-2-[(2-{[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl}ethoxy)methyl]propan-2-yl]carbamoyl}dodecanoate (4-c) [ka] Starting from 4-a, following general method B, compound 4-c was obtained as an off-white foamy solid (3.5 g, 94%).

[0064] Step 2: Preparation of 12-{[1,3-bis(2-{[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl}ethoxy)-2-[(2-{[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl}ethoxy)methyl]propan-2-yl]carbamoyl}dodecanoic acid (4-d) [ka] Starting from 4-c, following general procedure C, compound 4-d was obtained as an off-white foamy solid (1.8 g, 99%).

[0065] Example 5: [ka] Step 1: Preparation of methyl 12-({1,5-bis[(5-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentyl)carbamoyl]-3-{2-[(5-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentyl)carbamoyl]ethyl}pentan-3-yl}carbamoyl)dodecanoate (5-c) [ka] Starting from 5-a, following general procedure B, compound 5-c was obtained as an off-white foamy solid (4.0 g, 98%).

[0066] Step 2: Preparation of 12-({1,5-bis[(5-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentyl)carbamoyl]-3-{2-[(5-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentyl)carbamoyl]ethyl}pentan-3-yl}carbamoyl)dodecanoic acid (5-d). [ka] Starting from 5-c, following general procedure C, compound 5-d was obtained as an off-white foamy solid (1.5 g, 95%).

[0067] Example 6: [ka] Step 1: Preparation of 3-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}propanoic acid (6-b) [ka] Starting from 6-a, following general procedure A, compound 6-b was obtained as a white solid (10 g, 90%).

[0068] Step 2: Preparation of methyl 12-{[1,5-bis({[3-(3-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}propanamido)propyl]carbamoyl})-3-(2-{[3-(3-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}propanamido)propyl]carbamoyl}ethyl)pentan-3-yl]carbamoyl}dodecanoate (6-d) [ka] Starting from 6-b, following general method B, compound 6-d was obtained as a white solid (2.5 g, 82%).

[0069] Step 3: Preparation of 12-{[1,5-bis({[3-(3-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}propanamido)propyl]carbamoyl})-3-(2-{[3-(3-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}propanamido)propyl]carbamoyl}ethyl)pentan-3-yl]carbamoyl}dodecanoic acid (6-e) [ka] Starting from 6-d, following general procedure C, compound 6-e was obtained as an off-white foamy solid (0.8 g, 94%).

[0070] Example 7: [ka] Step 1: Preparation of methyl 12-{[1,5-bis({[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl})-3-(2-{[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl}ethyl)pentan-3-yl]carbamoyl}dodecanoate (7-c) [ka] Starting from 7-a, following general method B, compound 7-c was obtained as an off-white foamy solid (20.0 g, 93%).

[0071] Step 2: Preparation of 12-{[1,5-bis({[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl})-3-(2-{[2-(2-{[(2R,5R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}ethoxy)ethyl]carbamoyl}ethyl)pentan-3-yl]carbamoyl}dodecanoic acid (7-d) [ka] Starting from 7-c, following general procedure C, compound 7-d was obtained as an off-white foamy solid (15 g, 94%).

[0072] Thus, the present invention provides a convenient method for the preparation of glycolipid carboxylic acids in high yields by selective hydrolysis using common reagents and an environmentally friendly approach.

[0073] It should be understood that the present invention is susceptible to modifications, variations, and adaptations by those skilled in the art, and such modifications, variations, and adaptations are intended to be within the scope of the present invention.

Claims

1. Formula (I): 【Chemical 1】 〔wherein “n” ranges from 0 to 15; A １ , A ２ and A ３ are each, independently, H or 【Chemical 2】 selected from; However, A １ , A ２ and A ３ are not both H; G is the following group: 【Chemical Formula 3】 selected from; X is the following group 【Chemical Formula 4】 selected from; Y is 【Chemical Formula 5】 selected from; Z is the following group 【Chemical Formula 6】 selected from; R ２ is selected from an alkyl, aryl, alkoxy, aryloxy or arylalkyl group; “l”, “m” and “p” each independently range from 0 to 15〕 A method for producing a compound of, (a) In the presence of an enzyme selected from lipase, protease, amylase, trypsin, papain or a combination thereof and water, formula (Ia) [Chemical Formula 7] 〔wherein, R is selected from an alkyl, aryl or aralkyl group〕 of the compound is hydrolyzed; (b) The reaction mixture of step (a) is stirred, concentrated, dried and diluted with a solvent; (c) Removing the solid precipitate of the enzyme from the reaction mixture of step (b), purifying to obtain a compound of formula (I) A method comprising the steps

2. A １ 、 A ２ and A ３ each independently 【Chemical Formula 8】 wherein the substituent 【Chemical Formula 9】 is the following group: 【Table 1】 〔wherein R ２ is selected from an alkyl, aryl, alkoxy, aryloxy or arylalkyl group; “l”, “m”, “n” and “p” and “q” each independently range from 0 to 15〕 selected from, the method according to claim 1.

3. The method according to claim 1, wherein the enzyme in step (a) is present in an amount ranging from 1 to 100% by weight.

4. The method according to claim 1, wherein the enzymatic hydrolysis in step (a) is carried out at 20 to 45 ° C for 1 to 48 hours.

5. The method according to claim 1, wherein the solvent in step (b) is selected from water, methanol, ethanol, isopropyl alcohol, methyl tert-butyl ether, dichloromethane, ethyl acetate, acetone, dimethylformamide, tetrahydrofuran and acetonitrile or a combination thereof.

6. Formula (I’): 【Chemical 10】 〔wherein “n” ranges from 0 to 15; G is the following group: 【Chemical 11】 selected from; X is the following group 【Chemical 12】 selected from; Y is 【Chemical 13】 selected from; Z is the following group 【Chemical 14】 selected from; Here, R ２ is selected from an alkyl, aryl, alkoxy, aryloxy or arylalkyl group; “l”, “m” and “p” each independently range from 0 to 15〕 A method for producing a compound of, (i) A reactant selected from a glycolipid carboxylic acid of formula (z-b): 【Chemical Formula 15】 or a glycolipid amine compound of formula (z-bb): 【Chemical 16】 is coupled with an ester of formula (z-c): 【Chemical 17】 to form a compound of formula (Ia)’: 【Chemical Formula 18】 ; (ii) In the presence of an enzyme selected from lipase, protease, amylase, trypsin, papain or a combination thereof and water, hydrolyzing the ester compound of formula (Ia)’ to obtain a compound of formula (I)’, A method comprising the steps.

7. The method according to claim 6, wherein the coupling in step (i) is carried out in the presence of a coupling agent selected from hydroxybenzotriazole (HOBt), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (HBTU), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) or a combination thereof.

8. The method according to claim 6, wherein the coupling reaction in step (i) is carried out in the presence of a base selected from N,N-diisopropylethylamine (DIPEA).

9. The glycolipid carboxylic acid of formula (z-b) in step (i) is: In the presence of an enzyme and water, formula (z-a): 【Chemical 19】 The glycolipid carboxylic acid ester is hydrolyzed to obtain the glycolipid carboxylic acid of formula (z-b). The method according to claim 6, obtained thereby.

10. The compound of formula (I)' is 【Chemical 20】 【Chemical 21】 【Chemical 22】 Selected from, the method according to claim 6.

11. Formula (1-e) 【Chemical 23】 A method for producing a compound of, comprising: (i) In the presence of an enzyme and water, formula (1-a): 【Chemical 24】 The glycolipid carboxylic acid ester is hydrolyzed to Form a glycolipid carboxylic acid of formula (1-b): 【Chemical Formula 25】 ; (ii) The compound of formula (1-b) is coupled with an ester of formula (1-c): 【Chemical 26】 To form a compound of formula (1-d): 【Chemical 27】 ; (iii) In the presence of an enzyme and water, the ester compound of formula (1-d) is hydrolyzed to obtain the compound of formula (1-e). A method comprising the steps.

Images