US0093.76369B2 (12) United States Patent (10) Patent No.: US 9,376,369 B2 Chatterjee (45) Date of Patent: Jun. 28, 2016 (54) ANTI-CHOLESTEROLEMIC COMPOUNDS OTHER PUBLICATIONS AND METHODS OF USE Muruganandam et al. Indian Journal of Experimental Biology, vol. 40, Oct. 2002, pp. 1151-1160.* (71) Applicant: Subroto Chatterjee, Columbia, MD Wang et al..Journal of Ethnopharmacology 96 (2005) 483-487, pub (US) lished online Nov. 11, 2004.* Conner et al. Nutrition 12:274-277, 1996.* (72) Inventor: Subroto Chatterjee, Columbia, MD Definition of Inflammation by Medical dictionary, http://medical (US) dictionary.thefreedictionary.com/Inflammation, internet article downloaded Sep. 22, 2015.* - r Chae, Hypertension, 2001; 38:399-403.* (*) Notice: Subject to any disclaimer the term of this Abe I et al. "Potent and selective inhibition of squalene epoxidase by past list: G adjusted under 35 synthetic galloyl esters.” Biochem Biophys Res Commun. Apr. 2, M YW- y U days. 2000:270(1): 137-40. Alzheimer's Disease—downloaded Jun. 2011 from PubMed Health. (21)21) AppAppl. No.: 13/946,5589 American Heart Association, Cholesterol Abnormalities & Dis ease—downloaded Jun. 2011. (22) Filed: Jul.19, 2013 Baker, State University of New York at Buffalo Reporter, vol. 28, No. 35: Jul 24, 1997. O O Crespy Vetal. "A review of the health effects of green tea catechins (65) Prior Publication Data in in vivo animal models.” JNutr. Dec. 2004;134(12 Suppl):3431S US 2014/0031304 A1 Jan. 30, 2014 344OS. s Definition of food—dlownloaded from biology-online.org <http:// biology-online.org>, Jun. 2011. O O Merck Manuals Online: Stroke—downloaded Jun. 2011. Related U.S. Application Data Murase Tetal. “Gallates inhibit cytokine-induced nuclear transloca (63) Continuation of application No. 12488,965, filed on EEEEEE Jun. 22, 2009, now Pat. No. 8,492.351, which is a 1999; 19(6): 1412-20 ve. continuation of application No. PCT/US2007/088780, Muruganandam AV et al. “Effect of poly herbal formulation, EuMil, filed on Dec. 24, 2007. on chronic stress-induced homeostatic perturbations in rats.” Indian J Exp Biol. Oct. 2002:40(10): 1151-60. Abstract Only. (60) Provisional appli cation No. 60/876,761, filed on Dec. Okabe Set al. “New TNF-alpha releasing inhibitors, geraniin and 22, 2006, provisional application No. 60/876,599, corilagin, in leaves of Acer nikoense, Megusurino-ki.” Biol Pharm filed on Dec. 22, 2006, provisional application No. Bull. Oct. 2001:24(10): 1145-8. 60/877,753, filed on Dec. 29, 2006, provisional Okuda et al. In Phenolic Compounds in Food and Their Effects on application No. 60/877,740, filed on Dec. 29, 2006. Health II; ACS Symposium Series; American Chemical Society: Washington, DC, 1992. (51) Int. Cl. Tongia et al. Indian Journal of Indigenous Medicines, Oct. 1995; A2.3L I/30 (2006.01) 17(2): 13-24, abstract only. A6 IK3I/235 (2006.01) Kim et al. Influence of amla (Emblica officinalis Gaertn.) on A6 IK3I/7024 (2006.01) hypercholesterolemia and lipid peroxidation in cholesterol-fed rats. J A6 IK 45/06 (2006.01) Nutr Sci Vitaminol (Tokyo). Dec. 2005:51(6):413-8. CD7C 69/84 2OO 6. O1 Mathur et al. Hypolipidaemic effect of fruit juice of Emblica C7H 3/08 3:08: officinalis in cholesterol-fed rabbits. J Ethnopharmacol. Feb. 1996:50(2):61-8. (52) U.S. Cl. Thakur et al. The Ayurvedic medicines Haritaki, Amala and Bahira CPC ............... C07C 69/84 (2013.01); A23L I/3002 reduce cholesterol-induced atherosclerosis in rabbits. Int J Cardiol. (2013.01); A61 K3I/235 (2013.01); A61 K Nov. 1988:21(2):167-75. 31/7024 (2013.01); A61K 45/06 (2013.01); Zhang et al. Antiproliferative activity of the main constituents from C07H 13/08 (2013.01); A23 V 2002/00 Phyllanthus emblica. Biol Pharm Bull. Feb. 2004:27(2):251-5. (2013.01) k . (58) Field of Classification Search cited by examiner Nes lication file f 1 hhi Primary Examiner — Layla Berry ee application file for complete search history. (74) Attorney, Agent, or Firm — Mintz Levin Cohn Ferris (56) References Cited y and Popeo, P.C.; Peter F. Corless, Esq.; Daniel W. U.S. PATENT DOCUMENTS (57) ABSTRACT 4. E.ww. R. E. E.O.E. Thecholesteremic present invention activity providesfrom crude novel Embilica compounds officinialis with hypot (EO) FOREIGN PATENT DOCUMENTS R R methods of use. The invention also provides UaC1CalS. WO WO-2004/058213 A1 T 2004 WO WO 20060225O2 A1 * 3, 2006 ........... A61K 31,216 13 Claims, 11 Drawing Sheets U.S. Patent US 9,376,369 B2 HOOH OOH.HOOHOHOOH OHOOHOOOH OOH HO HO HOHOHOHOOH OOOSLOÀ? OHOHOOH HOOHHO HOOoHoºsºººo U.S. Patent Jun. 28, 2016 Sheet 3 of 11 US 9,376,369 B2 MONTHS FG. 2 MONTHS FIG. 3 U.S. Patent Jun. 28, 2016 Sheet 4 of 11 US 9,376,369 B2 200 10 O MONTHS FG. 4 U.S. Patent Jun. 28, 2016 Sheet 5 of 11 US 9,376,369 B2 22 S 22222222 S 2 S 22 SS 22. S D 222222222222222 S 2 C C C C c O C N r- y pifu 2 SS 2 S 22 S Z2 S ZZZZZZZZZZZZZZZ S 222222222222 N C C 3. w v pfu U.S. Patent Jun. 28, 2016 Sheet 6 of 11 US 9,376,369 B2 Cholest, 100 NChol, oleate N Ø FIG 6 U.S. Patent US 9,376,369 B2 SOALL Cd eSeld eqo 9, U.S. Patent Jun. 28, 2016 Sheet 8 of 11 US 9,376,369 B2 y c c CN co O cN r g 3 O O C O C O O C N. O w r N w s s (%) uoquoieselolo U.S. Patent Jun. 28, 2016 Sheet 9 of 11 US 9,376,369 B2 140 120 100 80 60 40 20 O Control LOW 2 3 4 4-1 5 6 7 8 7+8 Compounds FG. 9 21030 O O low 1 2 3 4 4+1 5 6 7 8 7-8 Compounds F.G. 10 U.S. Patent Jun. 28, 2016 Sheet 10 of 11 US 9,376,369 B2 2OO 150 1OO 5 O O Control Low 1 2 3 4 4-1 5 6 7 8 7-8 Compounds F.G. 11 U.S. Patent Jun. 28, 2016 Sheet 11 of 11 US 9,376,369 B2 SOAuu e Seleco. 9 US 9,376,369 B2 1. 2 ANT-CHOLESTEROLEMC COMPOUNDS opment of competitive inhibitors for 3-hydroxy-3-methylglu AND METHODS OF USE taryl coenzyme A reductase, a major regulatory enzyme of cholesterol biosynthesis. Many attempts to use for this pur RELATED APPLICATIONS pose oxygenated Sterols, which via binding oxysterol recep tors were expected to decrease activity of HMG-CoA reduc This application is a continuation of U.S. application Ser. tase, did not bring practical results. A series of fungal No. 12/488.965, filed Jun. 22, 2009 (now U.S. Pat. No. 8,492, metabolites with very high affinities for HMG-CoA reductase 351), which is a continuation of International Application No. were found to be highly efficient inhibitors of cholesterol PCT/US2007/088780 (WO 2008/080162) filed Dec. 24, biosynthesis. These compounds and Some synthetic analogs 2007, which claims the benefit of U.S. Provisional Applica 10 are commonly known as statins, are available commercially, tion 60/876,761 filed Dec. 22, 2006, U.S. Provisional Appli and are widely used. cation 60/877,753 filed Dec. 29, 2006, U.S. Provisional Although being relatively safe and efficient in treatment Application 60/876,599 filed Dec. 22, 2006 and U.S. Provi and prevention of coronary heart disease, statins have certain sional Application 60/877,740 filed Dec. 29, 2006. The entire 15 limitations in their use because of possible deleterious side contents of the aforementioned applications are hereby incor effects, such as muscle weakness, and renal failure. Statin porated herein by reference. therapy is contra-indicated in pregnant women and patients GOVERNMENT SUPPORT with liver disorders. Additionally, there is a significant patient population in whom statins are not effective, and so the need This invention was made with support from the United for an agent that will lower cholesterol extends beyond the States Government under grant DK-31771 and DK-31722. number of patients currently taking statins to lower their The US Government has certain rights in this invention. cholesterol levels. Taking additionally into account the rela tively high costs of statin therapy, which varies from $20,000 FIELD OF THE INVENTION to $40,000 per quality-adjusted life-year saved John A. 25 Farmer, Economic Implications of Lipid-Lowering Trials: The present invention provides novel compounds with Current Considerations in Selecting a Statin, Am J Cardiol hypocholesteremic activity from Embilica officinalis (EO) 1998; 82:26 M-31M, and the desirability of long-term per extracts and methods of use. The invention provides nutra manent treatment Terry A. Jacobson, Clinical Context: Cur ceuticals. The invention relates to the determination of the rent Concepts of Coronary Heart Disease Management, Am J. biological activity and mechanism of action by which the 30 Med. 2001; 110 (6A):3S-11S, it is evident that new agents purified compounds lower cholesterol levels. are needed with similar targeting as the statins but with higher potency, safety, and availability and that will provide signifi BACKGROUND OF THE INVENTION cant cost savings. Accordingly, the instant invention provides novel com According to the American Heart Association, an esti 35 pounds with anticholesterolemic activity, and methods of use. mated 100,870,000 American adults have total cholesterol levels in the borderline-high risk range of 200 mg/dl to 239 SUMMARY mg/dl., and there are 40,600,000 American adults living with high-risk cholesterol levels of 240 mg/dl or more.