US 20150209377A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0209377 A1 Lin et al. (43) Pub. Date: Jul. 30, 2015

(54) USE OF NOVEL MONOSACCHARIDE-LIKE filed on Sep. 13, 2014, provisional application No. GLYCYLATED SUGAR ALCOHOL 62/054,981, filed on Sep. 25, 2014. COMPOSITIONS FOR DESIGNING AND DEVELOPNG ANT-DABETC DRUGS Publication Classification (71) Applicants: Shi-Lung Lin, Arcadia, CA (US); (51) Int. Cl. Samantha Chang-Lin, Arcadia, CA A613 L/702 (2006.01) (US); Yi-Wen Lin, Arcadia, CA (US); A63L/22 (2006.01) Donald Chang, Arcadia, CA (US) (52) U.S. Cl. CPC ...... A6 IK3I/7012 (2013.01); A61 K3I/221 (72) Inventors: Shi-Lung Lin, Arcadia, CA (US); (2013.01) Samantha Chang-Lin, Arcadia, CA (US); Yi-Wen Lin, Santa Fe Spring, CA (57) ABSTRACT (US); Donald Chang, Cerritos, CA (US) This invention is related to a novel Sugar-like chemical com position and its use for diabetes therapy. Particularly, the (73) Assignees: Shi-Lung Lin, Arcadia, CA (US); present invention teaches the use of monosaccharide-like gly Samantha Chang-Lin, Arcadia, CA cylated Sugar alcohol compounds to block or reduce Sugar (US); Yi-Wen Lin, Arcadia, CA (US); absorption in diabetes patients, so as to prevent the risk of Donald Chang, Arcadia, CA (US) hyperglycemia symptoms. Glycylation of Sugar alcohols is a totally novel reaction that has never been reported before. (21) Appl. No.: 14/585,978 Therefore, the novelty of the present invention is that for the first time glycylated Sugar alcohols not only was found but (22) Filed: Dec. 30, 2014 also was found to be useful for treating Diabetes mellitus. In addition, the present invention teaches a method for produc Related U.S. Application Data ing these glycylated Sugar alcohols. In Sum, the present inven tion includes not only a kind of novel Sugar-like chemical (63) Continuation-in-part of application No. 14/457.829, compositions and its use for treating diabetes but also a state filed on Aug. 12, 2014. of-the-art protocol and methodology for producing Such a (60) Provisional application No. 61/931,650, filed on Jan. novel composition via glycylation of Sugars and Sugar alco 26, 2014, provisional application No. 62/050,107, hols. Patent Application Publication Jul. 30, 2015 Sheet 1 of 6 US 2015/0209377 A1

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USE OF NOVEL MONOSACCHARDE-LIKE may not be treated with insulin, depending on the pathologi GLYCYLATED SUGAR ALCOHOL cal causes of insulin resistance. COMPOSITIONS FOR DESIGNING AND 0004 Current medications for type-II DM can be catego DEVELOPING ANT-DABETC DRUGS rized into five groups: first, drugs increase insulin production in pancreas. Such as Sulfonylureas, Meglitinides and Stita PRIORITY gliptin; second, drugs decrease Sugar production in liver, Such 0001. The present invention claims priority to the U.S. as Metformin; third, drugs increase Sugar excretion via urine, Provisional Application Ser. No. 61/931,650 filed on Jan. 26, Such as Canagliflozin, forth, drugs reduce insulin resistance 2014, which was entitled “Composition and Formulation of in fat tissues and muscles, such as Thiazolidinediones; and Sugar Alcohol-Based Complexes for Delivering Nucleic last, drugs delay and/or reduce Sugar absorption in intestines, Acid-Based Drugs. In Vitro and InVivo”. The present inven Such as alpha-glucosidase inhibitors. However, due to the tion also claims priority to the U.S. Provisional Application kinetic properties of these drugs directed against certain natu Ser. No. 62/050,107 filed on Sep. 13, 2014, which was ral metabolic pathways of the body, they often present various entitled “Novel Monosaccharide-like Glycylated Sugar Alco degrees of side effects, including hypoglycemia (Sulfony hol Compositions useful for Diabetes Therapy'. The present lureas, Meglitinides, Stitagliptin and alpha-glucosidase invention further claims priority to the U.S. patent application inhibitors), kidney failure (Metformin and Canagliflozin), Ser. No. 14/457,829 filed on Aug. 12, 2014, entitled “Novel heart failure (Metformin and Thiazolidinediones), gas Sugar Alcohol-Based Compositions for Delivering Nucleic trointestinal problems (Metformin and alpha-glucosidase Acid-Based Drugs. In Vivo and In Vitro’. The present appli inhibitors), edema and anemia (Thiazolidinediones). In some cation is a continuation-in-part (CIP) application of the U.S. worst cases, death has been reported. patent application Ser. No. 14/457,829 filed on Aug. 12, 2014, 0005 To prevent the lethal problems of kidney and heart entitled “Novel Sugar Alcohol-Based Compositions for failure, the best way of treating type-II diabetes is to delay and Delivering Nucleic Acid-Based Drugs In Vivo and In Vitro'. reduce Sugar absorption in intestines; yet, without losing the which are hereby all incorporated by reference as if fully set Supply of energy. The current anti-diabetic drugs of this kind forth herein. Such as alpha-glucosidase inhibitors function through inhibi tion of intestinal enzymes that convert carbohydrates into FIELD OF INVENTION monosaccharides and hence can block Sugar uptake in intes 0002 This invention generally relates to a novel sugar-like tines. Nevertheless, the undigested carbohydrates may then chemical composition and its use for diabetes therapy. Par interact with colon bacteria to cause gastrointestinal prob ticularly, the present invention teaches the use of monosac lems, such as flatulence and diarrhea. From the beneficial charide-like modified Sugar alcohol compositions to replace view, this side effect may help to reduce obesity; yet, the Sugars for absorption in patients, so as to treat diabetes mel drawback is that blocking Sugar uptake also means loss of litus. In order to mimic monosaccharides, these novel Sugar energy Supply to the body. As a result, overdose of alpha alcohol compositions are modified by a reaction called gly glucosidase inhibitors can lead to acute hypoglycemia in cylation and turned into glycylated Sugar alcohols. As a result, diabetes patients. the novelty of the present invention is to use these glycylated 0006 Recently, it is further noted that the use of sugars and Sugar alcohols for reducing the Sugar uptake in human intes Sugar Substitutes such as artificial Sweeteners may worsen the tines, Subsequently preventing the hyperglycemia symptoms Sugar intolerance conditions of Diabetes mellitus in patients of diabetes patients after meals. In addition, the present inven (Suez et al., (2014) Nature in press, doi:10.1038/na tion teaches for the first time that chemical compounds like ture13793). Sugar alcohols and Sugars can be modified by glycylation to 0007. In sum, there is currently no safe drug agent or sugar reduce their pathological effects in diabetes. Therefore, the substitute that can treat type-II DM while still maintaining the present invention includes not only a kind of novel Sugar-like Supply of normal body energy. Therefore, it is highly desir chemical compositions and its use for treating diabetes but able to have a novel anti-diabetic composition that is able not also a state-of-the-art protocol and methodology for produc only to reduce Sugar absorption in intestines but also to pro ing Such a novel composition via glycylation. vide an alternative energy source to the body. BACKGROUND SUMMARY OF THE INVENTION 0003 Diabetes mellitus (DM), also known as diabetes, is a group of metabolic diseases that cause long-lasting hyperg 0008 Stem cells are treasure boxes containing various lycemia symptoms in humans as well as Some other mam effective ingredients useful for designing and developing cos mals, such as mice. These hyperglycemia symptoms include medical, pharmaceutical and therapeutic drugs and/or appli high blood Sugar levels (i.e. >140 mg/dL), frequenturination, cations, including, but not limited by, inducing and enhancing increased thirst and hunger, and more severely diabetic body cell/tissue/organ regeneration, repairing/healing dam ketoacidosis and hyperosmolar coma. Long-term hypergly aged tissues and organs, rejuvenating aged cells/tissues?or cemia often leads to heart diseases, stroke, kidney failure, gans, treating degenerative diseases (i.e. cancers, diabetes, ulcers in extremities, eye damages and obesity. There are two osteoporosis, Parkinson's and Alzheimer's diseases ... etc), types of diabetes: type-I DM results from lack of sufficient and preventing tumor/cancer formation, progression and/or insulin production in pancreas (or called insulin-dependent metastasis. In view of these potentials, the inventors had DM; IDDM), and type-II DM results from body’s failure to developed a novel technology to generate microRNA-in respond to insulin (or called insulin resistance; non-insulin duced pluripotent stem cells (iPSCs) in early 2008 (Lin et al., dependent DM; NIDDM). For treatment, type-I DM must be 2008, 2010 and 2011) and then successfully used these iPSCs managed with insulin injections while type-II DM may or as a tool for screening, identifying, isolating, and producing US 2015/0209377 A1 Jul. 30, 2015

potential drug ingredients for clinical testing and use (Chen DETAILED DESCRIPTION OF THE PREFERRED and Lin, (2013) Recent Patents on Regenerative Medicine 3, EMBODIMENT 5-16). 0013. As shown in FIGS. 2A-2C, a novel chemical reac 0009 Sugar alcohols are a generic kind of polyol alcohols tion glycylation of Sugar alcohols—has been developed to derived from Sugars and also frequently called polyhydric produce MGG, DGG and TGG. Previously, glycylation was alcohol, polyalcohol, or glycitol; Sugar alcohols have been only found to occur between amino acids but never in Sugar found to be abundantly glycylated in early stage iPSCs as alcohols or Sugars. Yet, as defined here, glycylation is a well. As defined in polymer chemistry, polyols are com chemical reaction that replaces the hydroxyl (HO—) groups pounds with multiple active hydroxyl groups available for of a Sugar alcohol or Sugar with glycine’s glycyl organic reactions and polymeric polyols are usually in a form (NHCHCOO or NH"CHCOO ) groups and thus of polyethers or polyesters. Most Sugar alcohols are white, results in the formation of an ether (R—O—R) linkage water-soluble natural occurring materials that are often used between each OH-removed carbon of the sugar/sugar alcohol in the cosmetic, pharmaceutical and food industries as and the glycyl group. This reaction involves a condensation humectants, thickeners and Sweeteners. They are represented process via dehydration. For example, when glycerol (or by a general chemical formula H(HCHO), H, which is dif called glycerin) is used as a model of Sugar alcohol, glycyla ferent from sugars H(HCHO), HCO. Also, unlike sugars tion of glycerol will generate three kinds of reactive products: which tend to form ring structures, Sugar alcohols do not. Yet, one is 1-, or 2-, or 3-monoglycylglycerol (MGG: FIG. 2A), they can be dehydrated into cyclic ethers, e.g. Sorbitol can be another is 1.2-, or 2.3-, or 1,3-diglycylglycerol (DGG; FIG. dehydrated to isosorbide. The sugar alcohols differ in chain 2B), and the last is 1,2,3-triglycylglyceride (TGG; FIG. 2C). length and have one hydroxyl (OH) group attached to each In this model, glycylation can be a partial or completed reac carbon (C) molecule in the chain. They are further differen tion; for instance, MGG and DGG are partially glycylated tiated by their relative orientation (stereochemistry) of these products, while TGG is completely glycylated. Since MGG, OH groups; for example, mannitol and Sorbitol are isomers DGG and TGG are all positively charged under acidic con that share the same chemical formula CH3(OH) but are ditions, such as in the gastrointestinal (GI) system, and hence different in the orientation of the OH group on carbon 2 (C). possess high affinity to glucose transporters (GLUT) that The common types of Sugar alcohols include, but not limited absorb sugars into cells. As a result, the binding of MGG/ by, alditol, arabitol, erythritol, fucitol, galactitol, glycerol (or DGG/TGG to GLUT prevents sugar uptake via GLUT in the called glycerin), iditol, inositol, isomalt, lactitol, maltitol, body, particularly in the GI system. By the same token, simi mannitol, polyglycitol, Sorbitol, threitol, Volemitol and Xyli lar to Sugar alcohols, simple Sugars including glucose, fruc tol. tose, galactose. Sucrose and lactose may also be modified by 0010 Sugar alcohols can be separated into three major glycylation and thus useful for reducing Sugar uptake in dia categories: monoglycylated glycerol (MGG), diglycylated betes patients as well. glycerin (DGG), and triglycylated glyceride (TGG). Using 0014 Like natural Sugars, these glycylated Sugar alcohols high performance liquid chromatography (HPLC), we can can be absorbed by and into human cells via glucose trans isolate and collect pure MGG and partially mixed DGG and porters (GLUT) and/or GLUT-mediated endocytosis. For TGG (FIG. 1). Further stereochemical analysis has shown example, FIG. 3 shows that in human liver HepG2 cells the that the structure of MGG is similar to monosaccharides such majority of MGG is absorbed by GLUT2 also known as as glucose, fructose and galactose, while those of DGG and Solute carrier family 2 (facilitated glucose transporter), mem TGG resemble disaccharides. In view of such similarity in ber 2 (SLC2A2) through a protein conformational change structures, MGG may be used to replace monosaccharides for and transporting mechanism. Yet, some MGG and the major interfering with and reducing Sugar absorption in diabetes ity of DGG and TGG bind GLUT2 with high affinity and thus patients. block the GLUT2 conformational change that is required for 0011. The present invention identified and disclosed for sugar influx. In this alternative pathway, the cells will then the first time that novel Sugar-like glycylated Sugar alcohol activate another endocytosis mechanism to internalize compounds can be useful for treating Diabetes mellitus. Spe GLUT2 along with its bound MGG/DGG/TGG, leading to a cifically, it is an object of the present invention to provide a decrease of GLUT2 on cell Surface and hence reducing Sugar method for treating the hyperglycemia symptoms and obesity uptake into GI tract cells. problems of Diabetes mellitus, which comprises administer 0015 The detail metabolism of glycylated sugar alcohols ing a composition comprising a glycylated Sugar or glycy in cells, namely glycylglycerolysis’, is proposed in FIG. 3. lated Sugar alcohol, wherein the glycylated Sugar or glycy After absorbed by the cells, glycylated Sugar alcohols such as lated Sugar alcohol comes from at least one Sugar alcohol or at MGG will be first processed into dihydroxyacetone phos least one Sugar modified by the reaction of glycylation; phate (DHAP) and then turned into its isomer D-glyceralde wherein the at least one Sugar alcohol has a formula of hyde-3-phosphate (GADP). After that, glyceraldehyde H(HCHO), H, and the at least one sugar has a formula of phospahte dehydrogenase (GAPDH) converts GADP into H(HCHO), HCO. D-1,3-bisphosphoglycerade (1,3-BPG), which is subse 0012. The present invention to provide a composition for quently converted by phosphoglycerate kinase (PGK) into developing treatments for hyperglycemia and obesity of Dia 3-phosphoglycerate (3-PG), by phosphoglycerate mutase betes mellitus as well. The composition comprises a glycy (PGM) into 2-phosphoglycerate (2-PG), and then further by lated Sugar or glycylated Sugar alcohol, wherein the glycy enolase into phosphoenolpyruvate (PEP). Last, after PEP is lated Sugar or glycylated Sugar alcohol comes from at least converted into pyruvate by a kinase (PK), this end product— one Sugar alcohol or at least one Sugar modified by the reac pyruvate—can be finally utilized by the cells for generating tion of glycylation; wherein the Sugar alcohol has a formula of energy (via Krebs cycle or citric acid cycle), producing Sug H(HCHO), H, and the sugar has a formula of H(HCHO) ars (via gluconeogenesis), and being involved in other cellu HCO. lar metabolisms. As a result, these glycylated Sugar alcohols US 2015/0209377 A1 Jul. 30, 2015 not only can lower blood sugar levels in diabetes patients but further indicates another potential use of glycylated Sugar also can serve as an alternative energy source for preventing alcohols intreating obesity, in particular diabetes-related obe acute hypoglycemia that has often seen in overdose of other sity in vivo. anti-diabetic drugs, such as alpha-glucosidase inhibitors. 0019. In view of all above evidences, we have practically 0016 Since glycylated sugar alcohols function through enabled the use of a novel glycylated Sugar alcohol compo reduction of Sugar absorption in human GI systems, oral sition for not only treating the hyperglycemia symptoms and administration is the best way to deliver this kind of medicine. obesity problems in diabetes individuals but also preventing Also, because the raw materials to make glycylated Sugar the risk of hypoglycemia that is often seen in other anti alcohols are Sugar alcohols and glycine, both of which are diabetic drugs. As a result, this kind of new medicine by food Supplements, this drug has been tested and shown no competing against monosaccharides to block or reduce Sugar sign of toxicity or hypoglycemia up to 1 g in mouse ("/40 of the uptake in the GI system may lead to a safer and more effective body weight). Yet, at high dosage (>0.65g per mouse), mild treatment for diabetes patients. In particular, since it is noted to moderate diarrhea may be observed in 15%-20% of the that all materials used to produce these glycylated Sugar alco tested mice post-meal. To further test its drug effects in treat hol compositions are totally non-toxic and safe under Food ing diabetes and obesity, we had fed 1 mg (in 1 mL) of purified and Drug Administration (FDA) regulations, this new kind of MGG to each of C57BL/6J diet-induced diabetic mice (n-3) anti-diabetic drugs may also be used as a food Supplement for right before meal, while C57BL/6J mice fed with 1 mL of maintaining the daily care of diabetes patients. ddHO were served as controls (n=3). After drug administra tion, each mouse was fed with a high carbohydrate mixture of A. Definitions 4 mg glucose and 2 g Kaytee mouse food for a single meal. 0020. To facilitate understanding of the invention, a num Then, whole blood sugar levels were measured at 0, 0.5, 1. ber of terms are defined below: 1.5. 2, 2.5, 3, 3.5, and 4 hours (hr) post-meal, using a Bayer 0021 Treated Cell: a single or a plurality of human or Contour Next USB system. No water uptake during this animal cells selected from the group consisting of a Somatic 4-hour measurement period of time. cell, a tissue cell, a stem cell, a germ-line cell, a tumor cell, a 0017. The results of blood sugar level changes were shown cancer cell, and a combination thereof. in FIG. 4. For MGG-treated mice, the blood glucose levels 0022 Glycylation: a chemical reaction that replaces the were measured as 85, 91, 77 mg/dL (average 84.3+7.0 hydroxyl (HO—) groups of a Sugar alcohol or Sugar with mg/dL) at 0 hr, 135, 147, 121 mg/dL (average 134.3+13.0 glycine’s or glycylated amino acid's glycyl (NH-CHCOO– mg/dL) at 0.5 hr, 157, 151, 147 mg/dL (average 151.7+5.5 or NH-CHCOO ) groups and thus results in the formation mg/dL) at 1 hr, 155, 131, 158 mg/dL (average 148-13.4 of an ether (R—O—R) linkage between each OH-removed mg/dL) at 1.5 hr, 129, 137, 133 mg/dL (average 133+4.0 carbon of the Sugar alcohol/Sugar and the glycyl group of the mg/dL) at 2 hr. 97, 121, 101 mg/dL (average 106.3+12.0 glycine and/or glycylated amino acid. mg/dL) at 2.5 hr, 91, 89, 88 mg/dL (average 89.3+1.5 mg/dL) 0023 Pharmaceutical and Therapeutic Application: a bio at 3 hr, 93, 85, 83 mg/dL (average 87+5.0 mg/dL) at 3.5 hr, medical utilization, treatment method, device and/or appara and 88, 93, 85 mg/dL (average 88.7-4.0 mg/dL) at 4 hr, tus useful for diagnosis, stem cell generation, stem cell respectively. As all detected numbers are comparably similar research and/or therapy development, tissue/organ repair to those from normal persons after meal, we concluded that and/or rejuvenation, wound healing treatment, tumor Sup the treatment of glycylated Sugar alcohols such as MGG pression, cancer therapy and/or prevention, disease treat obviously can maintain normal-like blood Sugar levels after ment, diabetes and obesity treatment, drug production, and a high carbohydrate meal in diabetes individuals. On the other combination thereof. hand, for untreated control mice, the blood glucose levels 0024 Prokaryotes: a one-cell organism that lacks a dis were measured as 73, 79.88 mg/dL (average 80+7.5 mg/dL) tinct membrane-bound nucleus and has its genetic materials at 0 hr, 255, 167,315 mg/dL (average 245.7+74.0 mg/dL) at in the form of a continuous strand of DNA, such as bacteria 0.5 hr,375,267,415 mg/dL (average 345.7+74.0 mg/dL) at 1 and archaea. hr, 315,281,379 mg/dL (average 325+49.0 mg/dL) at 1.5 hr, 295, 285, 311 mg/dL (average 297-13.0 mg/dL) at 2 hr. 264: 0025. Eukaryote or Eukaryotic Cell: an one-cell or mul 284,315 mg/dL (average 287.7+25.5 mg/dL) at 2.5 hr, 171, tiple-cell organism whose cells contain a nucleus and other 222, 299 mg/dL (average 230.7+64.0 mg/dL) at 3 hr, 157, structures (organelles) enclosed within membranes, such as 199, 285 mg/dL (average 213.7+64.0 mg/dL) at 3.5 hr, and yeast, plant and animal cells. 143, 166, 213 mg/dL (average 173.7+35.0 mg/dL) at 4 hr, B. Compositions and Applications respectively. After we further compared the detected numbers between MGG-treated and untreated mice, the results clearly 0026. The present invention discloses a novel glycylated showed that the treatment of glycylated Sugar alcohols sig Sugar alcohol composition and its use for treating the hyper nificantly lowers the absorption of glucose into blood stream glycemia symptoms and obesity problems of Diabetes mel and hence Successfully prevents the hyperglycemia Symp litus, comprising at least a Sugar alcohol or Sugar modified by toms in diabetes animals in vivo. the reaction of glycylation. 0018. In addition, loss of body weight has also been 0027. In addition, the present invention also teaches a observed in MGG-treated mice but not controls. As shown in novel method of glycylation and its use for producing posi FIG. 5, diabetic C57BL/6J mice (n=3) treated with 1 mg of tively charged Sugar alcohol and Sugar compounds capable of MGG (in 1 mL ddHO) before every meal (3 high carbohy interacting with negatively charged materials such as cell drate meals per day) for about one month have been found to Surface receptors and transmembrane transporters via elec lose about 16% to 30% (average 23%) of body weights, trostatic affinity and/or structural similarity. whereas those controls (n-3) treated with only 1 mL ddHO 0028. In the present invention, the disclosure of glycylated gained average over 49% of body weights. Clearly, this result Sugar alcohol/Sugar compositions and the reaction of glycy US 2015/0209377 A1 Jul. 30, 2015

lation in Sugar alcohols and Sugars have been first reported in Rockville, Md.); hESC (human embryonic stem cells); and our claimed priority of U.S. Ser. No. 14/457,829 filed on Aug. iPSC (induced pluripotent stem cells). 12, 2014, entitled “Novel Sugar Alcohol-Based Composi tions for Delivering Nucleic Acid-Based Drugs InVivo and In 1. Production of Glycylated Sugar Alcohol Compositions Vitro”. Furthermore, the use of these glycylated sugar alco 0037 Although the natural mechanism of sugar/sugar hols and/or Sugars for treating diabetes and obesity is totally alcohol glycylation is unclear, we have developed a chemical novel and never been reported before. As the metabolism of protocol and method to artificially make glycylated Sugar these glycylated Sugar alcohols is revealed in the present alcohols and glycylated Sugars. First, a solution was prepared, invention for the first time, the knowledge disclosed will be containing 1.0 M glycerol or fructose and 0.9% (w/v) NaCl in very useful for developing medicines and/or food Supple ddHO. Then, glycine was added into the solution to reach a ments for treating related metabolic diseases, such as diabetes final concentration of 0.05-3.0 M, depending on the desired and obesity. concentration of final glycylated Sugar alcohols/sugars, and mixed well. After overnight incubation at 4°C. with constant BRIEF DESCRIPTION OF THE DRAWINGS agitation, the glycine-added solution was heated at a tempera 0029 Referring particularly to the drawings and examples ture over 45° C., preferably >94° C., till the water (HO) for the purpose of illustration only and not limitation, there is content to be almost completely vaporized. This dry-style illustrated: dehydration protocol usually generated about 35% of MGG and about 65% of DGG/TGG. Before use, the final concen 0030 FIG. 1 shows the separation of DGG/TGG from tration of these glycylated Sugar alcohols could be adjusted MGG using HPLC purification. simply by adding a desired amount of autoclaved ddHO to 0031 FIG. 2A shows the chemical reaction of glycylation dissolve the dry reaction products. Alternatively, in order to at moderate dehydration and the resulting monoglycylated increase the MGG production rate, we further designed a Sugar alcohols (MGG) thereof, including 1-monoglycylglyc wet-style dehydration protocol, using autoclave machine. erol, 3-monoglycylglycerol, and 2-monoglycylglycerin. The Different from the aforementioned, after mixing glycine with chemical formula for monoglycylglycerol is NHCHCO the glycerol and NaCl solution, the mixture was then placed (HCHO). Hor NH"CHCO(HCHO)H, depending on its pH into an autoclave machine and run in a dry sterilization cycle condition. at about 121° C. and 100 kPa. After autoclave cycle was 0032 FIGS. 2B and 2C show glycylation after high levels completed, the reaction would contain about 78% of MGG of dehydration and the resulting diglycylated (DGG) and and about 22% of DGG/TGG. Before use, the final concen triglycylated sugar alcohols (TGG), respectively. For tration of the glycylated Sugar alcohols could be adjusted by example, when glycerol (glycerin) is used as a model of Sugar adding a desired amount of autoclaved ddHO into the reac alcohol, two major kinds of highly glycylated products can be tion. The chemical structures of MGG, DGG and TGG are identified: one is diglycylglycerol (2B: DGG) and the other is shown in FIGS. 2A-2C. 1,2,3-triglycylglyceride (2C; TGG). 0033 FIG. 3 shows the proposed metabolism of glycy 2. Human Cell Culture and Treatment lated sugar alcohols such as MGG in cells. For example, due to structural similarity, MGG is absorbed via either GLUT2 0038 Human liver cell line HepG2 was obtained from mediated carrier transportation or endocytosis. After uptake ATCC and cultivated at 37° C. under 5% CO, according to by the cells, MGG is processed through a mechanism similar manufacturer's Suggestions. to glycolysis into pyruvate, which is then useful for other 3. High Performance Liquid Chromatography (HPLC) important biological metabolisms, such as Krebs cycle and Analysis gluconeogenesis. 0034 FIG. 4 shows the therapeutic effects of glycylated 0039. A reverse-phase HPLC method has been developed Sugar alcohol-containing anti-diabetic drugs on the hyperg for separating MGG from DGG and TGG, based on their lycemia symptoms of diabetic mice in vivo. (male, 8-week different charges. HPLC programs were run by an Ultimate 3000 HPLC machine (Thermo Scientific) with a DNA Pac old C57BL/6J, total n=6). PA-100 column (BioLC Semi-Prep 9x250 mm) at a flow rate 0035 FIG. 5 shows the effects of glycylated sugar alcohol of 3.6 mL/min. Starting buffer was 50 mM Tris-HCl (pH7.6) treatments on the changes of body weight of diet-induced and mobile buffer was 50 mM Tris-HCl (pH7.6) with 500 mM obesity mice in vivo (male, 8-week-old C57BL/6.J., total n=6). Sodium perchlorate. Signals of differently charged glycylated sugar alcohols were detected with an UV detector at 260 nm. EXAMPLES The results were shown in FIG. 1. 0036. In the experimental disclosure which follows, the following abbreviations apply: M (molar); mM (millimolar); 4. In Vivo Evaluation of Glycylated Sugar Alcohol-Treated um (micromolar); mol (moles); pmol (picomoles); gm Diabetic Mice (grams); mg (milligrams); Lug (micrograms), ng (nanograms); 0040. To test the drug effects of MGG in treating diabetes, L (liters); mL (milliliters); uI (microliters); C. (degrees we had fed 1 mg (in 1 mL) of purified MGG to each of Centigrade); RNA (ribonucleic acid); DNA (deoxyribo C57BL/6J diet-induced diabetic mice (male, 8-week-old, nucleic acid); dNTP (deoxyribonucleotide triphosphate); n=3) right before meal. C57BL/6J mice fed with 1 mL of PBS (phosphate buffered saline); NaCl (sodium chloride); ddHO were served as control (male, 8-week-old, n=3). After HEPES (N-2-hydroxyethylpiperazine-N-2-ethanesulfonic drug administration, each mouse was fed with a high carbo acid); HBS (HEPES buffered saline); SDS (sodium dodecyl hydrate mixture of 4 mg glucose and 2 g Kaytee mouse food sulfate); Tris-HCl (tris-hydroxymethylaminomethane-hy for a single meal. Then, whole blood Sugar levels were mea drochloride); ATCC (American Type Culture Collection, sured at 0,0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4 hours (hr) post-meal, US 2015/0209377 A1 Jul. 30, 2015

using a Bayer Contour Next USB detector system. Neither 2. The method as defined in claim 1, wherein said glycy food nor water was given to those mice 4-hour before treat lation is a chemical reaction in that at least one hydroxyl ment and 4-hour post-meal. Yet, during each meal, another (—OH) group of the Sugar or the Sugar alcohol is replaced by additional 2 mL of ddHO was given to each mouse. The at least one glycyl (NH-CHCOO– or NH"CHCOO-) results were shown in FIG. 4. group from a glycine or a glycylated amino acid. 3. The method as defined in claim 1, wherein said sugar 5. In Vivo Evaluation of Glycylated Sugar Alcohol-Treated alcohol or said Sugar is selected from the group consisting of Obesity Mice glucose, fructose, galactose, Sucrose, lactose, alditol, arabi tol, erythritol, fucitol, galactitol, glycerol (glycerin), iditol, 0041. To test the drug effects of MGG in treating obesity, inositol, isomalt, lactitol, maltitol, mannitol, polyglycitol, we had fed 1 mg (in 1 mL) of purified MGG to each of sorbitol, threitol, volemitol, xylitol, and a combination C57BL/6J diet-induced obesity mice (male, 8-week-old, thereof. n=3) right before meal, while C57BL/6J mice fed with 1 mL 4. The method as defined in claim 1, wherein said glycy of ddHO were served as controls (male, 8-week-old, n=3). lated Sugar or glycylated Sugar alcohol contains monogly After drug administration, each mouse was fed with a high cylglycerol (MGG). carbohydrate mixture of 4 mg glucose and 2 g Kaytee mouse 5. The method as defined in claim 4, wherein said food for a single meal. Each mouse was treated and fed three monoglycylglycerol has a chemical formula of NH-CHCO times per day for about one month. Water (ddHO) was pro (HCHO),H or NH, CHCO(HCHO).H. vided all time without limits. The results were shown in FIG. 6. The method as defined in claim 1, wherein said glycy 5. lated Sugar or glycylated Sugar alcohol is positively charged. 7. The method as defined in claim 1, wherein said glycy 6. Statistic Analysis lated Sugar or glycylated Sugar alcohol is further in Solution 0042 Any change over 10% of signal intensity was con and has a concentration ranged from 0.1 uM to 10 M. sidered as a positive result, which in turn was analyzed and 8. The method as defined in claim 1, wherein said glycy presented as mean+SE. Statistical analysis of data was per lated Sugar or glycylated Sugar alcohol competes against with formed by one-way ANOVA. When main effects were sig monosaccharides for intestinal absorption in vivo. nificant, the Dunnett's post-hoc test was used to identify the 9. The method as defined in claim 1, wherein said glycy groups that differed significantly from the controls. For pair lated Sugar or glycylated Sugar alcohol interferes with Sugar wise comparison between two treatment groups, the two absorption in intestines. tailed student t test was used. For experiments involving more 10. The method as defined in claim 1, wherein said glycy than two treatment groups, ANOVA was performed followed lated Sugar or glycylated Sugar alcohol is absorbed into cells by a post-hoc multiple range test. Probability values of p-0. via glucose transporters. 05 was considered significant. All p values were determined 11. The method as defined in claim 1, wherein said glycy from two-tailed tests. lated Sugar or glycylated Sugar alcohol is a food Supplement. 12. The method as defined in claim 1, wherein said glycy REFERENCES lated Sugar or glycylated Sugar alcohol reduces glucose absorption in vivo and prevents the hyperglycemia symptoms 0043 1. Chen SKJ and Lin SL. (2013) Recent patents on and obesity problems of Diabetes mellitus. microRNA-induced pluripotent stem cell generation. 13. The method as defined in claim 1, wherein said glycy Recent Patents on Regenerative Medicine 3, 5-16. lated Sugar or glycylated Sugar alcohol is useful for develop 0044 2. Lin SL, Chang D, Chang-Lin S. Lin C. H. Wu D ing anti-diabetic drugs and therapy. TS, Chen DT, and Ying SY. (2008) Mir-302 reprograms 14. The method as defined in claim 1, wherein said glycy human skin cancer cells into a pluripotent ES-cell-like lated Sugar or glycylated Sugar alcohol is useful for develop state. RNA 14, 2115-2124. ing anti-obesity drugs and therapy. 0045 3. Linet al. (2011). Regulation of somatic cell repro 15. The method as defined in claim 1, wherein said glycy gramming through inducible mir-302 expression. Nucleic lated Sugar or glycylated Sugar alcohol is useful for cosmetic, Acids Res. 39, 1054-1065. pharmaceutical and therapeutic applications. 0046 4. Lin SL, Chang D, Ying SY. Leu D, and Wu DT 16. A composition for use in developing treatments for S. (2010) MicroRNA miR-302 inhibits the tumorigenecity hyperglycemia and obesity of Diabetes mellitus, comprising of human pluripotent stem cells by coordinate Suppression a glycylated Sugar or glycylated Sugar alcohol, wherein the of CDK2 and CDK4/6 cell cycle pathways. Cancer Res. glycylated Sugar or glycylated Sugar alcohol comes from at 70,9473-9482. least one Sugar alcohol or at least one Sugar modified by the 0047 5. Suez et al., (2014) Artificial sweeteners induce reaction of glycylation; glucose intolerance by altering the gut microbiota. Nature wherein the sugar alcohol has a formula of H(HCHO), H. in press, doi:10.1038/nature13793. and the sugar has a formula of H(HCHO), HCO. 1. A method for treating the hyperglycemia symptoms and 17. The composition as defined in claim 16, wherein said obesity problems of Diabetes mellitus, comprising adminis glycylation is a chemical reaction in that at least one hydroxyl tering a composition comprising a glycylated Sugar or glycy (—OH) group of the Sugar or the Sugar alcohol is replaced by lated Sugar alcohol, wherein the glycylated Sugar or glycy at least one glycyl (NHCHCOO– or NH"CHCOO-) lated Sugar alcohol comes from at least one Sugar alcohol or at group from a glycine or a glycylated amino acid. least one Sugar modified by the reaction of glycylation; 18. The composition as defined in claim 16, wherein said wherein the at least one Sugar alcohol has a formula of Sugar alcohol or said Sugar is selected from the group con H(HCHO), H, and the at least one sugar has a formula of sisting of glucose, fructose, galactose, Sucrose, lactose, aldi H(HCHO), HCO. tol, arabitol, erythritol, fucitol, galactitol, glycerol (glycerin), US 2015/0209377 A1 Jul. 30, 2015

iditol, inositol, isomalt, lactitol, maltitol, mannitol, polygly 25. The composition as defined in claim 16, wherein said citol, sorbitol, threitol, volemitol, xylitol, and a combination glycylated Sugar or glycylated Sugar alcohol is absorbed into thereof. cells via glucose transporters. 19. The composition as defined in claim 16, wherein said 26. The composition as defined in claim 16, wherein said glycylated Sugar or glycylated Sugar alcohol contains glycylated Sugar or glycylated Sugar alcohol is a food Supple monoglycylglycerol (MGG). ment. 20. The composition as defined in claim 19, wherein said monoglycylglycerol has a chemical formula of NH-CHCO 27. The composition as defined in claim 16, wherein said (HCHO), Hor NH, CH,CO(HCHO).H. glycylated Sugar or glycylated Sugar alcohol reduces glucose 21. The composition as defined in claim 16, wherein said absorption in vivo and prevents the hyperglycemia symptoms glycylated Sugar or glycylated Sugar alcohol is positively and obesity problems of Diabetes mellitus. charged. 28. The composition as defined in claim 16, wherein said 22. The composition as defined in claim 16, wherein said glycylated Sugar or glycylated Sugar alcohol is useful for glycylated Sugar or glycylated Sugar alcohol is further in developing anti-diabetic drugs and therapy. solution and has a concentration ranged from 0.1 uM to 10 M. 29. The composition as defined in claim 16, wherein said 23. The composition as defined in claim 16, wherein said glycylated Sugar or glycylated Sugar alcohol is useful for glycylated Sugar or glycylated Sugar alcohol competes developing anti-obesity drugs and therapy. against with monosaccharides for intestinal absorption in V1VO. 30. The composition as defined in claim 16, wherein said 24. The composition as defined in claim 16, wherein said glycylated Sugar or glycylated Sugar alcohol is useful for glycylated Sugar or glycylated Sugar alcohol interferes with cosmetic, pharmaceutical and therapeutic applications. Sugar absorption in intestines. k k k k k