US006540654B2 (12) United States Patent (10) Patent N0.: US 6,540,654 B2 Levy et a1. (45) Date of Patent: Apr. 1, 2003 (54) CAROTENOID ESTERS FOREIGN PATENT DOCUMENTS (76) Inventors: Luis W. Levy, c/o IneXa, Industria DE 199 50 327 A1 4/2000 EXtractora C.A., PO. BOX 17-03-4581, W0 W0 98/ 45241 A2 10/1998 QlliiO (EC); Richard H. BIIlIlIIlgtOIl, OTHER PUBLICATIONS 17 Castle Road, Sandal Wake?eld West Yorkshire, WFZ 7LU (GB); Anthony 5- CheW, et al., “Effects of Lutein from Marigold Extract on Tabatlllik, 75 sheringham, Immunity and Growth of Mammary Tumors in Mice”, Queensmead, SI J0hI(1’S \;V00d Park, Anticancer Research, vol. 16, pp. 3689—3694, (1996). London, NW8 6RB GB (List continued on neXt page.) ( * ) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 Primary Examiner_])eb0rah D_ can U-S~C~ 154(k)) by0 days- (74) Attorney, Agent, or Firm—Akin, Gump, Strauss, Hauer & Feld, L.L.P. (21) Appl. No.. 10/081,085 (57) ABSTRACT (22) Filed: Feb. 22, 2002 _ _ _ Monoesters, diesters and polyesters are provided Wherem (65) Prior Publication Data both the acid-derived moiety and the alcohol-derived moiety Us 2002/0169334 A1 NOV 14 2002 of the esters are carotenoid compounds. The synthetic ester ' ’ linkage betWeen tWo or more carotenoids provides neW Related US Application Data compounds in vvhich the similar and complementary prop (60) Provisional application NO_ 60/270,817’ ?led on Feb 23’ ert1es of the'mdividual carotenoids are combined. The neW 2001’ and provisional application No_ 60/347,443’ ?led on all-carotenoid esters may be useful as antioxidants, thera Jan. 11, 2002. peutic agents, pigmenting ingredients in poultry feed or as (51) Int. c1.7 .............................................. .. c07c 57/00 Coloring algemls for PM Thilpolyestersl h“? tile pofien?al to (52) US. Cl. ..................... .. 584/224; 584/223; 584/229; a? as H1969“ 3%? “6151 W“ M19516 6 6cm“ ‘Lon uctancg 584/230 c aracter1st1cs.~ e'a -caroteno1 esters may e prepare _ from the esteri?cation of at least one hydroXy carotenoid Fleld of Search ............................... .. at least one carboxylic carotenoid, OI. Via the reaction of 584/229’ 230 the acid chloride of a carboXylic carotenoid With a hydroXy (56) References Cited carotenoid. Preferred hydroXy carotenoids include lutein, ZeaXanthin, cryptoXanthin, violaXanthin, carotene diol, U.S. PATENT DOCUMENTS hydroXy carotene, hydroXylycopene, alloXanthin and dehy drocryptoXanthin. Preferred carboXylic carotenoids include 2,768,198 A 10/1956 Marbet et 211. biXin, norbiXin, [3-apo-8-carotenoic acid, crocetin, diapoc 3,206,316 A 9/1965 Kliiui arotenoic acid, carboXylcarotene and aZafrin. 5,536,504 A 7/1996 Eugster et 211. 5,959,138 A 9/1999 Torres-Cardona et 211. 6,191,293 B1 2/2001 Levy 3 Claims, 2 Drawing Sheets BIXIN “4 CH, \ Hmoc \ \ \ \ \ \ \ \ coo H: LUTEI N Lutein Monobixinate US 6,540,654 B2 Page 2 OTHER PUBLICATIONS Humeau, et al., “Enzymatic Esteri?cation of BiXin by Landrum, et al., “Lutein, ZeaXanthin, and the Macular L—ascorbic Acid”, Bi0techn0l. Lett., 22:2, (2000) Abstract Pigment”, Archives of Biochemistry and Biophysics, 385:1, Only. pp. 28—40, (Jan. 1, 2001). Gann, et al., “LoWer Prostate Cancer Risk in Men With Krinsky, “The Biological Properties of Carotenoids”, Pure & Appl. Chem, 66:5, pp. 1003—1010, (1994). Elevated Plasma Lycopene Levels: Results of a Prospective Straub, “Key to Carotenoids”, Institute of Organic Chemis Analysis”, Cancer Research, vol. 59, pp. 1225—1230, (Mar. try University of Berne, 2”“ and Revised Edition, (1987). 15, 1999). ALAM, et al., “Fatty Acid Composition of the Xanthophyll Esters of Tagetes ercta Petals”, Lipids, 3:2, pp. 183—184, Ziegler, “Carotenoids, Cancer, and Clinical Trials”, Annals (Mar. 1968). of the New York Academy of Sciences, vol. 691: “Caro Partali, et al., “Stable, Highly Unsaturated G1ycerides— tenoids in Human Health”, pp. 111—119, (1993). Enzymatic Synthesis With a Carotenoic Acid”, Angew. Chem. Int. Ed. EngL, 35:3, pp. 329—331, (1996). Roth, “Current Chemotherapy and Infectious Diseases”, Larsen, et al., “Combination of Vitamin E With a Carotenoid: (Nelson, et al., Eds. American Society of Microbiology: ot—Tocopherol and Trolox Linked to [3—Apo—8‘—carotenoic Washington, DC, pp. 1503—1505, (1980). [Out of Print and Acid”, Chem. Eur J., 4:1, pp. 113—117, (1998). No Longer Available]. U.S. Patent Apr. 1, 2003 Sheet 1 0f 2 US 6,540,654 B2 ZEFDJ PEwEQEat:viz-Eggs; U.S. Patent Apr. 1, 2003 Sheet 2 0f 2 US 6,540,654 B2 £000}/ // <UOQ0m 0mmx/£0m /4 75mmE5575mm a0.5%263aaging of#6000)!‘ m6 uooum: US 6,540,654 B2 1 2 CAROTENOID ESTERS For example, the valerate, laurate, oleate, linoleate and CROSS-REFERENCE TO RELATED caproate esters of the carotenoids Zeaxanthin and isoZeax APPLICATIONS anthin have been described in US. Pat. No. 5,536,504 of Eugster, et al. Short chain organic acid diesters of lutein and This application claims the bene?t of US. Provisional Zeaxanthin have also been described in US. Pat. No. 5,959, Patent Applications Nos. 60/270,817, ?led Feb. 23, 2001 138 of Torres-Cardona, et al. and No. 60/347,443 ?led Jan. 11, 2002. Esters containing a carotenoid as the acid-derived moiety BACKGROUND OF THE INVENTION are also Well knoWn. Whereas many occur in nature (“Key The carotenoids are important natural products Which are to Carotenoids”), other esters of this type have been pre involved in the photosynthetic process of plants. Caro 10 pared by synthesis. For example, US. Pat. No. 2,768,198 of tenoids are used as nutritional supplements for animals and Marbet, et al. describes the preparation of esters of bixin and humans, as Well as in food colorants and cosmetics. norbixin With amyl alcohol, octyl alcohol and Vitamin A. Although carotenoids are important for health, animals and Finally, glycerides With a carotenoid acid have been humans cannot produce them, and these compounds must prepared by enZymatic catalysis (Partali et al,Angew. Chem thus be obtained through diet from fruits and vegetables. 15 Int. Ed. Engl. 35:329—330 (1996)). The reaction of [3-apo Several properties of the carotenoids make them impor 8‘-carotenoic acid With Vitamin E yields ot-tocopheryl-[3-8‘ tant for the health of both animals and humans. For example, carotenoate as described by Larsen et al (Chem. Eur. J. these compounds are antioxidants With important quenching 4:113—117 (1998)). Additionally, the enZymatic esteri?ca effects on free radicals. They protect living tissues against a tion of bixin using L-ascorbic acid, Which contains tWo variety of diseases, either directly or as immunopotentiators. alcoholic OH groups, as the alcohol moiety has been Additionally, carotenoids are involved in gap-junction com reported by Humeau et al (Bi0techn0l. Lett. 22:155—168 munication among living cells. Recent epidemiological evi (2000)). dence has suggested an inverse relationship betWeen the In summary, although many esters containing carotenoid consumption of fruits and vegetables With high carotenoid components are knoWn, all of the knoWn carotenoid esters content and the incidence of several types of cancers. contain a non-carotenoid component as the alcohol-derived Speci?cally, [3-carotene, lutein and lycopene have been moiety or the acid-derived moiety. shoWn to exhibit a cancer-prevention effect (M. M. MatheWs-Roth, Current Chemotherapy and Infectious Dis BRIEF SUMMARY OF THE INVENTION eases (J. D. Nelson and C. Grassi, Eds,Am. Soc. Microbiol., 30 This invention is directed to esters comprising at least one Washington D.C.:1503—1505(1980)); B. P. CheW et al,Anti alcohol-derived moiety and at least one acid-derived moiety, cancer Research 16:3689—3694 (1996); P. H. Gann et al, Wherein the at least one alcohol-derived moiety comprises a Cancer Res. 59:1225—1230 (1999)). hydroxy carotenoid selected from the group consisting of Additionally, several carotenoids, such as lutein and monohydroxy carotenoids, dihydroxy carotenoids, and poly Zeaxanthin, have speci?c functions in the retina of the eye to 35 hydroxy carotenoids, and Wherein the acid-derived moiety assure healthy vision in several animal species, including comprises a carboxylic carotenoid selected from the group humans (J. D. Landrum et al., Archives Biochem. Biophys. consisting of monocarboxylic carotenoids, dicarboxylic 385(1):28—40(2001)). Finally, some carotenoids have pro carotenoids, and polycarboxylic carotenoids. Preferred vitamin A activity, Whereas others control reproduction and hydroxy carotenoids include lutein, Zeaxanthin, fertility, upregulate the Connexin43 gene, decrease the risk 40 cryptoxanthin, violaxanthin, carotene diol, hydroxy of degenerative disease and prevent coronary heart disease carotene, hydroxylycopene, alloxanthin and dehydrocryp (N. Krinsky, Pure and Appl. Chem. 66(5):1003-1010 toxanthin. Preferred carboxylic carotenoids include bixin, (1994)). norbixin, [3-apo-8-carotenoic acid, crocetin, diapocarotenoic From a chemical standpoint, carotenoids may be classi acid, carboxylcarotene and aZafrin. ?ed by their functional groups into several categories (see 45 Key to Carotenoids, 2nd enlarged and revised edition, H. BRIEF DESCRIPTION OF THE SEVERAL Pfander et al, Birkhauser Verlag, Basel, 1987). These include VIEWS OF THE DRAWINGS hydrocarbon carotenoids such as [3-carotene and lycopene, The foregoing summary, as Well as the folloWing detailed monohydroxy carotenoids such as [3-cryptoxanthin, dihy description of preferred embodiments of the invention, Will droxy carotenoids such as lutein and Zeaxanthin, polyhy be better understood When read in conjunction With the droxy carotenoids such as [3,[3-carotene triol, epoxycaro appended draWings. For the purpose of illustrating the tenoids such as violaxanthin and antheraxanthin, carbonyl invention, there are shoWn in the draWings embodiments carotenoids such as echinenone, capsanthin, canthaxanthin Which are presently preferred. It should be understood, and astaxanthin, and carotenoid acids such as bixin and hoWever, that the invention is not limited to the precise crocetin.