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`
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`Patent Office
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`KR20020095553A Pharmaceutical composition for prevention or treatment of obesity hyperlipidemia or fatty liver containing
`inhibitors of isocitrate dehydrogenase activity
`
`Applicants: TG BIOTECH INC [KR]
`
`Inventors: HUH TAE RIN [KR],KO HO JIN [KR],LEE SU MIN [KR],PARK JIN U [KR]
`
`Classifications:
`IPC
`
`A61K31/19;
`
`(IPC1-7): A61K31/19;
`
`CPC
`
`A61 K31 /194 (KR); A61 P3/04 (KR); A61 P3/06 (KR);
`
`Priorities: KR20010033599A 2001-06-14
`
`Application: KR20010033599A-2001-06-14
`
`Publication: KR20020095553A-2002-12-27
`
`Published as: KR10044232281 ; KR20020095553A
`
`Pharmaceutical composition for prevention or treatment of obesity hyperlipidemia or fatty liver containing inhibitors of
`isocitrate dehydrogenase activity
`
`Abstract
`
`PURPOSE: A preventing or therapeutic agent for obesity, hyperlipidemia or fatty liver containing an isocitrate dehydrogenase
`activity inhibitor as an effective ingredient is provided which has excellent effects for reducing body weight and suppressing
`obesity and biosynthesis of neutral lipid and cholesterol. CONSTITUTION: This prophylactic or treating agent for obesity,
`hyperlipidemia or fatty liver contains oxalomalic acid, methylisocitric acid, oxaloacetate and glyoxylate as an isocitrate
`dehydrogenase activity inhibitor. This agent is administered to a patient at dosages in a preferred range of about 7mg to 70g
`per day as adult patient(70kg).
`
`1 of 1
`
`8/ 13/2022, 12:10 PM
`
`Rigel Exhibit 1031
`Page 1 of 28
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`

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`Rigel Exhibit 1031
`Page 2 of 28
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`Rigel Exhibit 1031
`Page 2 of 28
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`Rigel Exhibit 1031
`Page 3 of 28
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`Rigel Exhibit 1031
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`Rigel Exhibit 1031
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`Rigel Exhibit 1031
`Page 5 of 28
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`Rigel Exhibit 1031
`Page 5 of 28
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`Rigel Exhibit 1031
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`Rigel Exhibit 1031
`Page 8 of 28
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`Page 14 of 28
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`Rigel Exhibit 1031
`Page 14 of 28
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`

`

`~ Europiiisches
`
`Patent~mt
`Eu.ropcan
`Patent Office
`Office europeen
`des brevets
`
`Patent Translate
`
`Powered by EPO and Google
`
`Notice
`This translation is machine - generated. It cannot be guaranteed that it is intelligible,
`accurate, complete, reliable or fit for specific purposes. Critical decisions, such as
`commercially relevant or financial decisions, should not be based on machine -
`translation output.
`
`DESCRIPTION KR100442322B1
`11 Obesity, hyperlipidemia or fatty liver prevention or treatment comprising an isocitrate
`dehydrogenase activity inhibitor as an active ingredient {Pharmaceutical composition
`for prevention or treatment of obesity, hyperlipidemia or fatty liver containing
`inhibitors of isocitrate dehydrogenase activity}
`
`[ 1]
`18 1 is a graph showing the degree of inhibition of enzyme activity in adipocytes treated
`with oxalo malic acid and methyl isocitric acid, which are isocitric acid dehydrogenase
`activity inhibitors;
`
`[2]
`24 ♦ : 0.5mM oxalomalic acid, ■ : 0.5mM methylisocitric acid,
`
`[3]
`28 2 is a graph showing the inhibitory effect of fat accumulation in adipocytes treated
`with oxalo malic acid and methyl isocitric acid, which are isocitric acid dehydrogenase
`activity inhibitors;
`
`[4]
`34 3A is a photograph of the result of comparing the amount of fat accumulation in the
`back neck region in mice (right) administered with oxalomalic acid, an isocitric acid
`dehydrogenase activity inhibitor, with that of normal mice (left),
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`Page 15 of 28
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`

`[5]
`40 3B is a photograph of the result of comparing the amount of abdominal fat
`accumulation in mice (right) administered with oxalomalic acid, an isocitric acid
`dehydrogenase activity inhibitor, with that of normal mice (left),
`
`[6]
`46 3C is a photograph of the result of comparing the size of the abdominal adipose tissue
`from a mouse {right) administered with oxalomalic acid, an isocitric acid
`dehydrogenase activity inhibitor, with a normal mouse (left),
`
`[7]
`52 3D is a photograph of the result of comparing the degree of fat accumulation in the
`subcutaneous tissue in a mouse (right) administered with oxalomalic acid, an isocitric
`acid dehydrogenase activity inhibitor, with that of a normal mouse (left),
`
`[8]
`58 4 is a bar graph showing the effect of inhibiting the increase in body weight in mice
`administered with oxalo malic acid, an isocitric acid dehydrogenase activity inhibitor;
`
`[9]
`63 5 is a bar graph showing the effect of lowering blood triglycerides (triglycerides) in
`mice administered with oxalo malic acid, an isocitric acid dehydrogenase activity
`inhibitor;
`
`[ 10]
`69 6 is a bar graph showing the effect of lowering blood cholesterol in mice administered
`with an isocitrate dehydrogenase activity inhibitor.
`
`[ 11]
`74 The present invention relates to a therapeutic agent for obesity, hyperlipidemia or
`fatty liver comprising an isocitrate dehydrogenase activity inhibitor as an active
`ingredient, and more particularly, oxalomalic acid, methyl isocitric acid It relates to a
`preventive or therapeutic agent for obesity, hyperlipidemia and fatty liver comprising
`an isocitric acid dehydrogenase activity inhibitor, including (methylisocitric acid), as
`an active ingredient.
`
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`

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`[ 12]
`83 Isocitric acid dehydrogenase is a major enzyme in the tricarboxilic acid (TCA) cycle
`for energy metabolism. It oxidatively decarboxylates citric acid to produce a -
`ketoglutaric acid and NADH or NADPH. is known to do
`
`[ 13]
`89 The isocitrate dehydrogenase of higher animals is a mitochondrial NAD+ - specific
`isocitrate dehydrogenase (hereinafter abbreviated as 'IDH') , cytoplasmic NADP +,
`depending on the type of cofactor and its location in the cell. - specific isocitrate
`dehydrogenase (hereinafter abbreviated as 'IDPc') and mitochondrial NADP+ -
`specific isocitrate dehydrogenase (hereinafter abbreviated as 'IDPm') of three
`isoenzymes ( isozymes) .
`
`[ 14]
`98 Among them, IDH produces NADH and a - chitoglutarate by decarboxylating isocitric
`acid within the TCA cycle, which not only metabolizes energy through the electron
`transport system, but also biosynthesis of amino acids such as glutamic acid,
`glutamine, arginine and proline, or As it is an intermediate metabolite for synthesizing
`other biological metabolites, it acts as an important regulator of the TCA cycle for
`energy metabolism, protein biosynthesis, and nitrogen metabolism.
`
`[ 15]
`101 IDPm and IDPc show strong tissue specificity in terms of enzymatic activity. In
`cardiac tissue, more than 90% of the total enzymatic activity of NADP + - specific
`isocitrate dehydrogenase is present in mitochondria, and about 10% is present in the
`cytoplasm. Conversely, in liver tissue, only 3% exists in mitochondria and the
`remaining 97% is known to exist in the cytoplasm {Plaut, G.W.E, Current Topics in
`Cell Regulation, 2, 1 - 27, 1983).
`
`[ 16]
`116 For isoenzymes of isocitrate dehydrogenase, only the structural characteristics of
`the isoenzyme have been partially elucidated, but the functions have not been
`elucidated.
`
`[ 17]
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`122 On the other hand, fat accumulation in higher animals is made according to the
`following process.
`124 That is, when there is excessive energy in the body, differentiation of adipocytes,
`which increases the number and size of white adipose tissue, is promoted, and the
`accumulation of lipids is increased. Accordingly, expression of the ob gene in white
`adipose tissue is increased. An increase in the concentration of leptin in the body
`appears, thereby changing the action of hormones in the brain to decrease appetite.
`will go through
`130 In white adipose tissue, the expression of genes such as Peroxisome Proliferator(cid:173)
`Activated Receptor r (PPAR r ) , C/EBPa, and ADD 1 /SREBP 1, known as master
`transcription factors for adipocyte differentiation, is promoted to promote adipocyte
`differentiation and fat accumulation. is increased, and excess energy in the body is
`stored in the form of fat, thereby regulating the energy balance in the body (Hu, E.
`et al., Proc.
`13a Natl.
`137 Acad.
`13s Sci.
`139 USA, 1995, 92, 9856- 9860; Keller, H. et al., Proc. Natl. Acad. Sci. USA, 1993, 20,
`9856- 9860; Freytag. S.O. et al., Genes Dev., 1994, 8, 1654 - 1663; Tontonoz, P. et
`al., Mol. Cell. Biol., 1993, 13, 4753- 4759; Spiegelman, B. M., Cell, 1996, 87, 377 -
`389). Substances that act as ligands in vivo for the activation of PPAR r, the main
`transcription factor for adipocyte differentiation, are fatty acids such as linoleic acid,
`docosahexaenoic acid (DHA) , and arachidonic acid ( polyunsaturated fatty acids
`such as arachidonic acid (Krey, G. et al., Mol. Endocrinol., 1997, 11, 779- 791; Yu et
`al., J. Biol. Chem., 1995, 270, 23975 - 23983) together with prostaglandin 12
`(Forman B. M. et al., Cell, 1995, 83, 803- 812; Kliewer. S. A. et al., Cell, 1995, 83,
`813 - 819) are known.
`
`[ 18]
`152 The present inventors found that the increase in the gene expression and enzyme of
`isocitrate dehydrogenase, and the resulting increase in NADPH, which is an enzyme
`reaction product, increases the accumulation of fat, causing obesity and fatty liver,
`and at the same time, the triglyceride ( triglyceride) and cholesterol (cholesterol)
`were also confirmed to increase (Korea Patent Application 2000 - 0061962) .
`157 Accordingly, the present inventors have found that oxalomalic acid and
`methylisocitric acid, which are isocitric acid dehydrogenase activity inhibitors, are
`administered to adipocytes and mice, and have excellent weight loss and obesity
`inhibitory effects, as well as biological neutral lipid and cholesterol lowering effects.
`The present invention was completed by confirming that the isocitrate
`dehydrogenase activity inhibitor can be used as a preventive or therapeutic agent for
`obesity, hyperlipidemia or fatty liver.
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`[ 19]
`167 It is an object of the present invention to provide a preventive or therapeutic agent
`for obesity, hyperlipidemia, or fatty liver, which contains an isocitric acid
`dehydrogenase activity inhibitor as an active ingredient, which is excellent in a
`weight loss effect, an obesity suppression effect, and a bioneutral lipid and
`cholesterol lowering effect.
`
`[20]
`175 In order to achieve the above object, the present invention provides a preventive or
`therapeutic agent for obesity, hyperlipidemia, or fatty liver comprising an isocitric
`acid dehydrogenase activity inhibitor as an active ingredient.
`
`[21]
`181 In addition, the present invention provides a preventive or therapeutic agent for
`obesity, hyperlipidemia, or fatty liver, characterized in that it contains oxalo malic
`acid or methyl isocitric acid as an active ingredient.
`
`[22]
`187 Hereinafter, the present invention will be described in detail.
`
`[23]
`191 The present invention provides a preventive or therapeutic agent for obesity,
`hyperlipidemia, or fatty liver, comprising an isocitric acid dehydrogenase activity
`inhibitor as an active ingredient.
`
`[24]
`197 The present inventors found that NADPH production in the cytoplasm including IDPc
`gene, which is an isozyme of isocitrate dehydrogenase, increase in expression of
`isocitrate dehydrogenase gene and increase in enzymatic activity, and increase in
`NADPH is involved in lipid and cholesterol biosynthesis It was confirmed that it
`promotes various enzymatic reactions such as 0061962) .
`
`[25]
`205 In addition, the present invention provides a preventive or therapeutic agent for
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`obesity, hyperlipidemia, or fatty liver, characterized in that it contains oxalo malic
`acid or methyl isocitric acid as an active ingredient.
`
`[26]
`211 Oxalomalic acid is produced by the aldol condensation reaction of oxaloacetate, an
`intermediate of the TCA cycle, and glyoxylate, an intermediate of the glyoxylate
`cycle, in vivo, and specifically inhibits the activity of isocitric acid dehydrogenase,
`and Isocitric acid strongly inhibits isocitric acid dehydrogenase in competition with
`isocitric acid as a substrate.
`
`[27]
`219 When oxalomalic acid and methylisocitric acid are added to adipocytes, the activity of
`isocitric acid dehydrogenase is reduced by about 50% in 30 minutes and 1 hour,
`respectively (FIG. 1). In addition, when treated with preadipocytes, differentiation
`into adipocytes is inhibited, and the accumulation of fat in the adipocytes is also
`suppressed (FIG. 2). In this case, the concentration of oxalomalic acid and methyl
`isocitric acid is preferably in the range of 0.1 µ M to 100 mM, and preferably in the
`range of 10 µM to 5 mM.
`
`[28]
`229 It was confirmed through a mouse experiment that the isocitric acid dehydrogenase
`activity inhibitor showed a high effect on obesity suppression and triglyceride and
`cholesterol lowering.
`232 This is confirmed by measuring the following several indicators, such as measuring
`the increase in abdominal fat and the amount of lipid accumulation in the body.
`
`[29]
`237 The present inventors administered oxalomalic acid and methylisocitric acid to mice
`by intraperitoneal injection 3 times a week at 25 mg/kg of body weight for 14
`weeks.
`240 As a result, the body weight was reduced by more than 10% compared to the control
`mice administered intraperitoneally with physiological saline (FIG. 4). When the
`above mice were dissected, it was confirmed that the size of the epididymal fat pad
`was very small, and the weight of the adipose tissue was reduced by about 5 times.
`In addition, the dorsal view after removing the outer skin also showed that the
`amount of obese tissue in the mice treated with the isocitric acid dehydrogenase
`activity inhibitor was significantly reduced (FIG. 3). In addition, in order to explore
`the degree of fat accumulation in the subcutaneous tissue, the abdominal
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`

`subcutaneous tissue is cut in cross - section, and the tissue is embedded in paraffin
`after the usual fixation procedure and dehydration process. It can be confirmed that
`the trixane dehydrogenase inhibitor has an important effect on weight loss and
`reducing the amount of intracellular fat accumulation (Fig. 3).
`
`[30]
`255 The present inventors confirmed how the above inhibitors of isocitric acid
`dehydrogenase activity, such as oxalomalic acid and methylisocitric acid, affect lipids
`and cholesterol in the body.
`258 The amount of triglyceride and total cholesterol was measured by separating the
`serum of mice administered with oxalomalic acid and methylisocitric acid, which are
`the isocitric acid dehydrogenase activity inhibitors. As a result, compared to the
`control mice, 50 % and decreased by about 30% ( FIGS. 5 and 6 ) .
`
`[31]
`265 As described above, it can be confirmed that the isocitric acid dehydrogenase activity
`inhibitor has the effect of reducing body weight, effectively inhibits fat accumulation
`in the body, and inhibits the biosynthesis of cholesterol, thereby lowering the
`production amount of the living body cholesterol complex.
`
`[32]
`272 The effective dose of the prophylactic or therapeutic agent for obesity,
`hyperlipidemia, or fatty liver containing the isocitric acid dehydrogenase activity
`inhibitor of the present invention as an active ingredient is usually about 7 mg to 70
`g per day for an average adult patient (70 kg) . is the range of
`
`[33]
`279 The specific dosage of the preventive or therapeutic agent for obesity,
`hyperlipidemia or fatty liver containing the isocitric acid dehydrogenase activity
`inhibitor of the present invention as an active ingredient depends on age, weight,
`symptoms, disease to be treated, administration route, treatment period, etc. it is
`decided
`284 That is, the doctor will determine the most appropriate actual dosage for each
`patient, and the actual dosage may vary depending on the patient's age, weight,
`symptoms, and disease to be treated.
`287 Accordingly, the dosage may be lower or greater than the above - mentioned ranges,
`and all such dosages are within the scope of the present invention.
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`[34]
`292 The prophylactic or therapeutic agent for obesity, hyperlipidemia or fatty liver
`containing the isocitric acid dehydrogenase activity inhibitor of the present invention
`as an active ingredient may be administered alone, but in general, a pharmaceutical
`selected from the intended route of administration and standard pharmaceutical
`practice. It is administered in admixture with an acceptable carrier, or diluent.
`297 The administration of the preventive or therapeutic agent for obesity, hyperlipidemia,
`or fatty liver of the present invention may be made in a form for oral administration
`such as a solid dosage form, a liquid dosage form or other dosage form, or a form for
`parenteral administration such as an injection solution, a coating agent or a
`suppository.
`302 In addition, the preventive or therapeutic agent for obesity, hyperlipidemia, or fatty
`liver of the present invention may be administered once to several times a day.
`
`[35]
`307 The prophylactic or therapeutic agent for obesity, hyperlipidemia, or fatty liver using
`the isocitric acid dehydrogenase activity inhibitor of the present invention as an
`active ingredient may be administered systemically or locally through oral or
`parenteral administration.
`311 For example, it is administered orally in the form of tablets, pills, granules, capsules,
`solutions, emulsions, suspensions, syrups, and the like. When administered by a
`parenteral method, for example, in the case of an injection, it may be injected
`intravenously, intramuscularly or subcutaneously. For example, it may be applied to
`the skin in the form of a patch, ointment, or topical preparation for transdermal
`administration, and may be administered intrarectally in the form of a suppository.
`Typical solid dosage forms for oral administration include powders, tablets, pills,
`granules, capsules, etc., wherein the active ingredients are mixed with one or more
`inert diluents (eg, starch, cellulose, lactose, kaolin, etc.) commonly used in the art. It
`is mixed, and may be mixed with any lubricant, binder, disintegrant, stabilizer, and
`the like. Typical liquid formulations for oral administration include pharmaceutically
`acceptable solutions, emulsions, suspensions, syrups, elixirs, and the like, wherein
`the active ingredients are contained in one or more inert diluents (eg, purified water,
`ethanol, etc.) commonly used in the art. and may be mixed with any sweetening
`agent, flavoring agent, preservative, and the like. Typical injections for parenteral
`administration include sterile solutions, emulsions, suspensions, and the like,
`wherein the active ingredient