WO2007097652A2 - Synthetic mannosylglycerate derivatives for the stabilisation and preservation of biomaterials - Google Patents
Synthetic mannosylglycerate derivatives for the stabilisation and preservation of biomaterials Download PDFInfo
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- WO2007097652A2 WO2007097652A2 PCT/PT2007/000012 PT2007000012W WO2007097652A2 WO 2007097652 A2 WO2007097652 A2 WO 2007097652A2 PT 2007000012 W PT2007000012 W PT 2007000012W WO 2007097652 A2 WO2007097652 A2 WO 2007097652A2
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
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- the present invention concerns the use of synthetic Mannosylglycerate derivatives, like mannosyl-Lactate or manosyl-glycerol, alone or as constituents of a formulation to protect and/or stabilise enzymes or other cellular components, or other materials with biologically derived components, against deleterious agressions, namely caused by high temperatures, high osmolarity, free-radicals, desiccation, freeze-drying, storage and/or repetitive use, thereby improving the processes, sensitivity, and/or shelf-life.
- synthetic Mannosylglycerate derivatives like mannosyl-Lactate or manosyl-glycerol
- thermophilic and hyperthermophilic micro-organisms have been acknowledged as a source of new compounds usable for several biotechnological applications. Among them, is the use of novel, highly efficient and compatible solutes for the preservation of biomaterials.
- solutes like, di- myoinositol-phosphate, di-mannosyl-di-myo-inositol-phosphate, diglycerol phosphate, mannosylglucosylglycerate, mannosylglycerate, and mannosylglyceramide, have been identified in these organisms and the intracellular content of these solutes responds or reacts to stress conditions, such as overoptimal osmolarity or temperature, presumably to lessen the aggressive effects.
- mannosylglycerate has been the most extensively studied and shown to protect enzymes and proteins, in vitro, far better than the commonly used compatible solutes [1-3).
- mannosylglycerate is a well-known bio- stabiliser with thermophilic origin, whose industrial uses are protected under a European patent application [4],
- nucleic acid molecules like DNA, or RNA
- stability of nucleic acid molecules can be increased by the addition of compatible solutes from hyperthermophiles, such as described for ectoins [5], and their use in several applications in medicine, pharmaceutical industry, or scientific research can be envisioned.
- This improvement can have as a target: to gain proteins presenting a higher additional stability; the improved enzyme performance under more aggressive conditions of temperature, pressure, ionic strength, pH, detergent or organic solvents presence; the stabilization and/or protection against denaturation of immobilized proteins, nucleic acids, cellular membranes or liposomes protection of proteins, nucleic acids, cell membranes, liposomes or cells towards; the deleterious effects of desiccation ; or the protection of general biomaterials against loss of function or viability during its transport, storage or frequent use.
- mannosyl-lactate has a higher stabilising effect on protein structure when compared with other protein stabilizers.
- mannosyl-Lactate is capable of elevating the melting temperature of staphylococcal nuclease A (SNase) of the egg's white lysozyme of chicken more effectively than its natural homologue, mannosylglycerate, being therefore more effective in protecting these proteins against thermal denaturation and consequent deactivation.
- SNase staphylococcal nuclease A
- Recombinant SNase was produced in Escherichia coli cells (strain HBlOl) and purified by osmotic shock and cationic chromatography.
- the protein stock solution was prepared by successive dialysis at 4°C in sodium phosphate buffer (1OmM, pH 7.5). A protein final concentration of approximately 0.02 mM was used on the Differential Scanning Calorimetry (DSC) assays.
- the chicken's egg white lysozyme was commercially obtained (Fluka) as a lyo- philized powder, and used without further purification.
- a protein stock solution was prepared after successive dialysis at 4°C in sodium citrate buifer (2OmM, pH 6.0).
- a protein final concentration of approximately 0.025 mM was used on the Differential Scanning Calorimetry (DSC) assays.
- the compatible solutes used in this example were potassium mannosylglycerate, potassium di-myo-inositol phosphate, ectoin, hydroxyectoine, trehalose and glycerol.
- the solutes were all used at a final concentration of 500 mM.
- Mannosyl-lactate and mannosyl-glycerol were chemically synthesized while mannosylglycerate was extracted and purified from Rhodothermus obamensis cells.
- Di- myo-inositol phosphate, ectoin and hydroxyectoine were commercially obtained (Bitop). The purity and quantification of these compounds was assessed by NMR.
- mannosyl-lactate is capable of elevating the melting temperature of staphylococcal nuclease A to a greater extent than its natural homologue mannosylglycerate on a larger temperature range, and can therefore be used to protect proteins in general against thermal denaturation and consequent deactivation effectively to a large concentration range. Furthermore, it is capable of the same stabilizing effect as its natural homologue mannosylglycerate to lower concentrations.
- the pig's heart malate dehydrogenase was commercially obtained (Roche) in an aqueous solution containing 50% of glycerol and extensively dialysed, in a sodium phosphate buffer 50 mM (pH 7.6).
- the enzyme (0.2mg/mL) was incubated at 20 0 C in a thermostatisized cell, in the presence or absence of 500 mM of mannosylglycerol or mannosyl-lactate, in a spectrofluorimeter (SPEX-Fluorologl680, Edison , NJ , USA) and measured the light dispersion at 320 nm.
- Figure 1 Represents the generic chemical structure of mannosylglycerate synthetic derivatives mentioned in the present invention, in all its possible stereoisomeric forms.
- the figure is intended to represent all hexoses either in the ⁇ or in the b configuration.
- the letters 1 Rl' and 'R2' are intended to represent a carboxylate, a methyl, an amide, or a primary alcohol group.
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Abstract
The present invention consists in the utilization of synthetic mannosylglycerate derivatives, such as mannosyl-lactate or mannosylglicerate, or mixtures thereof alone or as constituents in a suitable formulation viewing the protection and/or stabilization of enzymes, proteins, antibodies, DNA or RNA molecules, biological membranes, liposomes, or other cellular components and biomaterials against aggressive conditions to the same.
Description
Description
SYNTHETIC MANNOSYLGLYCERATE DERIVATIVES FOR THE STABILISATION AND PRESERVATION OF BIO- MATERIALS
Field of the Invention
[1] The present invention concerns the use of synthetic Mannosylglycerate derivatives, like mannosyl-Lactate or manosyl-glycerol, alone or as constituents of a formulation to protect and/or stabilise enzymes or other cellular components, or other materials with biologically derived components, against deleterious agressions, namely caused by high temperatures, high osmolarity, free-radicals, desiccation, freeze-drying, storage and/or repetitive use, thereby improving the processes, sensitivity, and/or shelf-life.
[2] The referred compounds can be described as hexoses involved in a glycosidic bond to a linear three-carbon compound, and obey to the general formula depicted in Figure 1. Prior State of the Art
[3] In recent years, thermophilic and hyperthermophilic micro-organisms have been acknowledged as a source of new compounds usable for several biotechnological applications. Among them, is the use of novel, highly efficient and compatible solutes for the preservation of biomaterials. Several new and unusual solutes like, di- myoinositol-phosphate, di-mannosyl-di-myo-inositol-phosphate, diglycerol phosphate, mannosylglucosylglycerate, mannosylglycerate, and mannosylglyceramide, have been identified in these organisms and the intracellular content of these solutes responds or reacts to stress conditions, such as overoptimal osmolarity or temperature, presumably to lessen the aggressive effects.
[4] Among these compounds, mannosylglycerate has been the most extensively studied and shown to protect enzymes and proteins, in vitro, far better than the commonly used compatible solutes [1-3). The use of mannosylglycerate as a stabilising agent of biomaterials has already been disclosed, in fact, mannosylglycerate is a well-known bio- stabiliser with thermophilic origin, whose industrial uses are protected under a European patent application [4],
[5] The enhanced protein stability rendered by certain low-molecular wheight organic solutes allows enzymes to function under more severe conditions of temperature, pressure, ionic strength, pH, presence of detergents or organic solvents. One of the priorities of modern biotechnology is to obtain stable enzymes or agents that stabilise those enzymes against thermal or chemical denaturation. The ability of some compatible solutes to stabilise enzymes is, therefore, of great importance to modern bi-
otechnology. This point is obviously extended to all proteins, that are used or can be used in processes where their stability is necessary, since all proteins either with or without enzymatic activity share the same overall basic elements of structure and may be protected against denaturation or inactivation through the same general mechanisms or processes.
[6] Also, it should be emphatized that compatible solutes are capable of protecting proteins, cell membranes, lipossomes, and cells from the deleterious effects of desiccation, and possess strong moistening properties. The preservation of desiccated or Iyophilized cells or cell components and biomaterials has many applications in medicine, pharmaceutical industry, cosmetic industry, food industry, and scientific research. In spite of the great importance of desiccation and freezing in the conservation of biological samples, acertain degree of denaturation of proteins or a decrease of the viable count of cell cultures inevitably takes place during these processes of preservation, and could be prevented or diminished by the use of low molecular wheight stabilisers.
[7] Also, the stability of nucleic acid molecules, like DNA, or RNA, can be increased by the addition of compatible solutes from hyperthermophiles, such as described for ectoins [5], and their use in several applications in medicine, pharmaceutical industry, or scientific research can be envisioned. Description
[8] What originated the present invention was verifying that, although mannosylglycerate is an outstanding stabilizer, it is possible to improve such abilities through the chemical synthesis of analogs designed for that purpose. This is the case for mannosyl-lactate, a synthetic derivative of mannosylglycerate, not found in nature and which contains a single hydroxyl group substitution by a hydrogen atom on the glycerate's position 3. This simple substitution confers the new compound an improved performance as an enzyme or nucleic acids stabilizer. Therefore, it is logical to assume that similar replacements will produce equal or bigger improvements.
[9] Thus, and according to the disclosed state of the art, with increased effectiveness of the biomaterial stabilizers progress, through mannosylglycerate derivatives chemical synthesis will be (according to the examples), a considerable breakthrough in the biomaterial stabilizing area, as well as in all industrial and/or commercial procedures where the biomaterial stability is taken into account.
[10] This way, the usage of synthetic mannosylglycerate derivatives, such as mannosyl- lactate, mannosylglycerol, glucosyl-lactate or glucosylglyceramide, among others, like additives in adequate formulations, will lead to an improvement of processes using materials of biologic origin. This improvement can have as a target: to gain proteins presenting a higher additional stability; the improved enzyme performance under more
aggressive conditions of temperature, pressure, ionic strength, pH, detergent or organic solvents presence; the stabilization and/or protection against denaturation of immobilized proteins, nucleic acids, cellular membranes or liposomes protection of proteins, nucleic acids, cell membranes, liposomes or cells towards; the deleterious effects of desiccation ; or the protection of general biomaterials against loss of function or viability during its transport, storage or frequent use.
[11] The synthetic derivatives of mannosylglycerate can be added in orders of concentration of mM, to preparations of biological compounds, improving its performance or life span in a more effective way than other natural compounds. Example 1
[12] . This example shows that mannosyl-lactate has a higher stabilising effect on protein structure when compared with other protein stabilizers. In particular, mannosyl-Lactate is capable of elevating the melting temperature of staphylococcal nuclease A (SNase) of the egg's white lysozyme of chicken more effectively than its natural homologue, mannosylglycerate, being therefore more effective in protecting these proteins against thermal denaturation and consequent deactivation.
[13] Recombinant SNase was produced in Escherichia coli cells (strain HBlOl) and purified by osmotic shock and cationic chromatography. The protein stock solution was prepared by successive dialysis at 4°C in sodium phosphate buffer (1OmM, pH 7.5). A protein final concentration of approximately 0.02 mM was used on the Differential Scanning Calorimetry (DSC) assays.
[14] The chicken's egg white lysozyme was commercially obtained (Fluka) as a lyo- philized powder, and used without further purification. A protein stock solution was prepared after successive dialysis at 4°C in sodium citrate buifer (2OmM, pH 6.0). A protein final concentration of approximately 0.025 mM was used on the Differential Scanning Calorimetry (DSC) assays.
[15] The compatible solutes used in this example were potassium mannosylglycerate, potassium di-myo-inositol phosphate, ectoin, hydroxyectoine, trehalose and glycerol. In addition to these solutes, the effect of mannosylglycerol and of the potassium salts of mannosyl-lactate, inorganic phosphate and chloride was also examined. The solutes were all used at a final concentration of 500 mM.
[16] Mannosyl-lactate and mannosyl-glycerol were chemically synthesized while mannosylglycerate was extracted and purified from Rhodothermus obamensis cells. Di- myo-inositol phosphate, ectoin and hydroxyectoine were commercially obtained (Bitop). The purity and quantification of these compounds was assessed by NMR.
[17] DSC assays were performed on a MicroCal VP-DSC Microcalorimeter. After degassing under vacuum for 8 minutes, samples were heated from 25°C to 950C with a heating rate of l°C/min.
[18] Mannosyl-lactate ( 500 mM ) proved to be an outstanding stabilizer elevating the melting temperature of SNase and Iysozyme by 10.00C and 7.4°C respectively (figures 2 and 3), while, under the same conditions, the natural homologue mannosyl glycerate is only able to increase the respective melting temperatures by 7.1°C and 5.7°C. None of the tested solutes was able to surpass the stabilizing effect of mannosyl-lactate. Example 2
[19] This example shows that mannosyl-lactate is capable of elevating the melting temperature of staphylococcal nuclease A to a greater extent than its natural homologue mannosylglycerate on a larger temperature range, and can therefore be used to protect proteins in general against thermal denaturation and consequent deactivation effectively to a large concentration range. Furthermore, it is capable of the same stabilizing effect as its natural homologue mannosylglycerate to lower concentrations.
[20] The Staphylococcal nuclease A was produced and purified as described in example
1, just like the protein stock solution and the DSC assays were performed as described in example 1.
[21] The effect of the concentration of mannosyllactate on the melting temperature of
SNase was studied up to 1 M. The result is shown in Figure 4 and compared to the result obtained in the presence of mannosylglycerate it is shown that mannosyl-lactate can elevate the melting temperature of Snase better than mannosylglycerate up to concentrations at least as high as 1 M. Example 3
[22] This example shows that mannosyl-lactate and mannosylglycerol are capable of strongly decrease the performance of the protein assembly degree in the pig's heart malate dehydrogenase.
[23] The pig's heart malate dehydrogenase was commercially obtained (Roche) in an aqueous solution containing 50% of glycerol and extensively dialysed, in a sodium phosphate buffer 50 mM (pH 7.6). The enzyme (0.2mg/mL) was incubated at 200C in a thermostatisized cell, in the presence or absence of 500 mM of mannosylglycerol or mannosyl-lactate, in a spectrofluorimeter (SPEX-Fluorologl680, Edison , NJ , USA) and measured the light dispersion at 320 nm. At the beginning of the experiment, the temperature was quickly raised up to 400C and the light dispersion due to the protein assembly was monitored for a long period of time. The results presented in figure 5, clearly show the ability of these two solutes to decrease the extent of the protein assembly of the enzyme. Brief Description of the Figures
[24] Figure 1 . Represents the generic chemical structure of mannosylglycerate synthetic derivatives mentioned in the present invention, in all its possible stereoisomeric forms.
The figure is intended to represent all hexoses either in the α or in the b configuration. The letters 1Rl' and 'R2' are intended to represent a carboxylate, a methyl, an amide, or a primary alcohol group.
[25] Figure 2. SNase melting temperature in the presence or absence of different solutes added at 500 mM concentration.
[26] Figure 3. Lysozyme melting temperature in the presence or absence of different solutes added at 500 mM concentration.
[27] Figure 4. SNase melting temperature as a function of mannosyl-glycerate (dotted line) and mannosyl-lactate (full line) concentration.
[28] Figure 5. Malate dehydrogenase assembly monitored by light dispersion at 320nm in the presence or absence of mannosyl-glycerol and mannosyl-lactate, added at a concentration of 50OmM. References
[29] [1] Ramos, A., N. D. H. Raven, R. J. Sharp, S. Bartolucci, M. Rossi, A. Cannio, J.
Lebbink, J. van der Oost, W. M. de Vos and H. Santos. 1997. Stabilisation of enzymes against thermal stress and freeze-drying by mannosylgiycerate. Appl. Environ. Microbiol. 63:4020-4025.
[30] [2] Borges, N., A. Ramos, N. D. H. Raven, R. J. Sharp, and H. Santos. 2002. Comparative study of the thermostabilising properties of mannosylgiycerate and other compatible solutes on model enzymes. Extremophiles 6:209-216.
[31] [3] Lamosa, P., A. Burke, R. Peist, R. Huber, M. Liu, G. Silva, C. Rodrigues-
Pousada, J. LeGail, C. Maycock, and H. Santos. 2000. Thermostabilisation of proteins by diglycerol phosphate, a new compatible solute from the hyperthermophile Ar- chaeo globus fulgidus. Appl. Environ. Microbiol. 66:1974-1979.
[32] [4] Santos , H., A. Ramos, M. S. da Costa. Thermostabilisation, osmoprotection, and protection against desiccation of enzymes, cell components and cells by mannosylgiycerate. European Patent no. 97670002.1.
[33] [5] Malin G, Iakobashvili R, and Lapidot A (1999) Effect of tetrahydropyrimdine derivatives on protein-nucleic acids interaction. Type II restriction endonucleases as a model system. J. Biol. Chem. 274:6920-6929.
Claims
Claims
[I] The utilization of synthetic mannosylglycerate derivatives, namely mannosyl-lactate or mannosylglycerol, comprising all its possible ste- reoisomeric forms, alone or as a constituent in a suitable formulation, characterized in that it confers increased protection or stability to enzymes, other cellular components or other biomaterials in stress conditions, wherein thereby improving its performance, sensitivity or shelf-life of the mentioned materials.
[2] The utilization of synthetic mannosylglycerate derivatives according to claim 1, wherein a three-carbon polar group replaces the glycerate residue. [3] The utilization of synthetic mannosylglycerate derivatives according to claims 1 and 2, wherein the glycerate residue is replaced by a lacryl group. [4] The utilization of synthetic mannosylglycerate derivatives according to claims 1 and 2, wherein the glycerate residue is replaced by a glyceryl group. [5] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 4, wherein the three-carbon polar residue may be in the D or L configuration. [6] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 5, wherein the glycosidic bond is established between position
1 of the hexose and position 3 of the three-carbon polar residue. [7] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 5, wherein the glycosidic bond is established between position
1 of the hexose and position 2 of the three-carbon polar residue. [8] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 5, wherein the glycosidic bond is established between position
1 of the hexose and position 1 of the three-carbon polar residue. [9] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 8 wherein the mannose residue is replaced by any other hexose, namely glucose, galactose, gulose, talose, fucose, rhamnose, idose, or altrose. [10] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 9, wherein the hexose is in the α or β configuration.
II 1] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 10, wherein conferring protection to enzymes or other proteins against thermal denaturation and/or induced aggregation by purification, transport repeated use and/or storage.
[12] The utilization of synthetic mannosylgly cerate derivatives according to claims 1 to 11, wherein it confers protection to the activity of polymerase chain reaction (PCR) enzymes for clinical, biological and industrial purposes or research and development during the high-temperature recycling of the enzymes as well as its transport or storage.
[13] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 11, wherein it confers stability to enzymes or other proteins during lyophilization, desiccation and storage at low temperatures.
[14] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 11, wherein it confers additional stability during the manufacture, storage and usage of test kit enzymes, for diagnostic, industrial or research and development purposes.
[15] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 11, wherein it confers additional stability to enzymes or other proteins during their routine utilization for diagnostic, industrial or research and development purposes.
[16] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 11, wherein it suppresses or reduces the extension of the protein assembly for therapeutic, diagnostic, industrial or research and development purposes.
[17] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 10, wherein it confers structural protection and/or additional stabilisation towards a thermal attack or enzymatic degradation of DNA or RNA molecules.
[18] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 10, wherein it confers structural protection and/or additional stability to cell membranes and lipossomes, subjected to stress conditions such as desiccation and/or lyophilization.
[19] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 10, wherein it confers additional stability improves moisturising properties of cosmetic products stabilises compounds within lipossomes or suppresses of oxygen free radicals.
[20J The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 10, wherein it confers additional stability to antibodies during its use for clinical, academic or industrial purposes.
[21] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 10, wherein it confers additional stability to vaccines during its use for clinical, academic or industrial purposes.
[22] The utilization of synthetic mannosylglycerate derivatives according to claims 1 to 10, wherein it confers additional protection to microbial cells against the damage caused by such processes as lyophilization, desiccation, high temperatures, or freezing.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PT103442A PT103442A (en) | 2006-02-24 | 2006-02-24 | MANOSYL-GLYCERATE SYNTHETIC DERIVATIVES FOR THE STABILIZATION AND / OR PRESERVATION OF BIOMATERIALS |
| PT103442 | 2006-02-24 |
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| WO2007097652A2 true WO2007097652A2 (en) | 2007-08-30 |
| WO2007097652A3 WO2007097652A3 (en) | 2008-03-06 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104561107A (en) * | 2013-10-16 | 2015-04-29 | 镇江拜因诺生物科技有限公司 | Method for improving biogas yield through mannosyglycerate |
| CN104560946A (en) * | 2013-10-10 | 2015-04-29 | 镇江拜因诺生物科技有限公司 | Application of glycerolglycerate in PCR as synergist |
| CN104543378A (en) * | 2013-10-16 | 2015-04-29 | 镇江拜因诺生物科技有限公司 | Application of phytase stabilizer in feed processing |
| WO2015137838A1 (en) * | 2014-03-14 | 2015-09-17 | Extremochem, Lda. | Hexose derivatives, preparation and uses thereof |
| WO2019092504A1 (en) * | 2017-11-13 | 2019-05-16 | Extremochem, Lda. | Neutral glycosylated amides and dianionic glucuronidated acids as stabilizers for biological molecules |
| CN111793664A (en) * | 2020-06-19 | 2020-10-20 | 中国科学院天津工业生物技术研究所 | Method for biologically preparing mannose glyceric acid |
| CN116251110A (en) * | 2022-11-18 | 2023-06-13 | 广东药科大学 | Application of beta-D-glucopyranosyl-lactic acid and derivatives thereof in preparation of medicines for treating metabolic diseases and inflammation |
| US12098162B2 (en) | 2018-11-13 | 2024-09-24 | Extremochem, Lda | Neutral glycosylated amides and dianionic glucuronidated acids as stabilizers for biological molecules |
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2006
- 2006-02-24 PT PT103442A patent/PT103442A/en not_active Application Discontinuation
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| KR20160145584A (en) * | 2014-03-14 | 2016-12-20 | 파쿠우다지 지 씨엔씨아스 다 우니베르시다지 지 리스보아 | Hexose derivatives, preparation and uses thereof |
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| CN111683957A (en) * | 2017-11-13 | 2020-09-18 | 埃克斯特雷莫凯姆有限公司 | Neutral glycosylated amides and dianionic glucuronidated acids as stabilizers for biomolecules |
| WO2019092504A1 (en) * | 2017-11-13 | 2019-05-16 | Extremochem, Lda. | Neutral glycosylated amides and dianionic glucuronidated acids as stabilizers for biological molecules |
| US12098162B2 (en) | 2018-11-13 | 2024-09-24 | Extremochem, Lda | Neutral glycosylated amides and dianionic glucuronidated acids as stabilizers for biological molecules |
| CN111793664A (en) * | 2020-06-19 | 2020-10-20 | 中国科学院天津工业生物技术研究所 | Method for biologically preparing mannose glyceric acid |
| CN116251110A (en) * | 2022-11-18 | 2023-06-13 | 广东药科大学 | Application of beta-D-glucopyranosyl-lactic acid and derivatives thereof in preparation of medicines for treating metabolic diseases and inflammation |
Also Published As
| Publication number | Publication date |
|---|---|
| PT103442A (en) | 2007-08-31 |
| WO2007097652A3 (en) | 2008-03-06 |
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