US008551507 B2

(12) United States Patent (10) Patent No.: US 8,551,507 B2 Liu (45) Date of Patent: Oct. 8, 2013

(54) TERPENE GLYCOSIDES AND THEIR Paeoniae Radix, and Increase of Water Solubilty of COMBINATIONS AS SOLUBLIZINGAGENTS Proanthocyanidins in the Presence of Paeoniflorin. Chem. Pharm. Bull 48(2), 2000, pp. 201-207.* (75) Inventor: Zhijun Liu, Baton Rouge, LA (US) Anne Marie Fine, Oligomeric Proanthocyanidin Complexes: His tory, Structure, and Phytopharmaceutical Applications. Alternative (73) Assignee: Board of Supervisors of Louisiana Medicine Review, vol. 5 No. 2, 2000, pp. 144-151.* State University And Agricultural and AMA Drug Evaluations Annual, “Clofazimine Lamprene. pp. Mechanical College, Baton Rouge, LA 1619-1620 (1993). (US) Eicher, Tet al., “Azole.” in The Chemistry of Heterocycles. Structure, Reactions, Syntheses, and Applications, Wiley-VCH. (2003). (*) Notice: Subject to any disclaimer, the term of this Kimata, H. et al., “Interaction of of Bupleuri Radix with patent is extended or adjusted under 35 Ginseng Saponin: Solubilization of Saikosaponin-a with Chikusetsu U.S.C. 154(b) by 0 days. saponin V (=Ginsenoside-Ro).” Chem. Pharm. Bull. (Tokyo), vol.33, pp. 2849-2853 (1985). Lipinski, C.A., “Drug-like Properties and the Causes of Poor Solu (21) Appl. No.: 13/379,072 bility and Poor Permeability.” J Pharm Tox Meth, vol. 44, pp. 235-249 (2000). (22) PCT Filed: Jun. 24, 2010 Lipinski, C., et al., “Experimental and Computational Approaches to Estimate Solubility and Permeability in Drug Discovery and Devel (86). PCT No.: PCT/US2010/039799 opment Settings.” Adv. Drug Deliv. Rev., vol. 23, iss. 1-3, pp. 3-25 (1997). S371 (c)(1), Material Safety Data Sheet, “Artemisinin”. (2), (4) Date: Jan. 25, 2012 Material Safety Data Sheet, “'. Material Safety Data Sheet, “Silybin ()”. (87) PCT Pub. No.: WO2010/151653 Sasaki, Y. et al., “Solubilizing Properties of and Its PCT Pub. Date: Dec. 29, 2010 Derivatives: Solubilization of Saikosaponin-a, the Saponin of Bupleuri Radix.” vol. 36, No. 9, pp. 3491-3495 (1988). (65) Prior Publication Data Tanaka, T. et al., “New Monoterpene Glycoside Esters and Penolic Constsituents of Paeoniae Radix, andlincreaase of Water Solubility of US 2012/O121696A1 May 17, 2012 Proanthocyanidins in the presence of Paeoniflorin.” Chem. Pharm. Bull., vol. 48, No. 2, pp. 201-207 (2000). Tanaka, T. et al., “Relationship between Hydrophobicity and Struc Related U.S. Application Data ture of Hydrolyzable Tannins, and Association of Tannins with Crude (60) Provisional application No. 61/219,973, filed on Jun. Drug Constituents in Aqueous Solution.” Chem. Pharm. Bull., vol. 24, 2009. 45, No. 12, pp. 1891-1897 (1997). Tanaka, T. et al., “Rubusoside (b-D-glucosyl ester of 13-O-b-D- (51) Int. Cl. glucosyl-Steviol), a Sweet principle of Rubus chingii Hu A6 IK3I/7048 (2006.01) (Rosacease).” Agricultural and Biological Chemistry, vol. 45, No. 9. pp. 2165-2166 (1981). A6 IK3I/337 (2006.01) The Merck Index, 12" Edition, Ed. S. Budavariet al., pp. 282-283 A6 IK3I/7034 (2006.01) (Camptothecin); 287-288 (); 450 (Curcumin); 737 (Gallic C7H 5/24 (2006.01) Acid); 894-895 (Itraconazole); and 1428 (Rutin) (1996). (52) U.S. Cl. USPC ...... 424/400; 424/777: 514/25; 514/449; * cited by examiner 514/450 (58) Field of Classification Search Primary Examiner — Frederick Krass None Assistant Examiner — Michael P Cohen See application file for complete search history. (74) Attorney, Agent, or Firm — John H. Runnels: Bonnie J. Davis (56) References Cited (57) ABSTRACT U.S. PATENT DOCUMENTS Several terpene glycosides (e.g., mogroside V, paenoiflorin, 4,943,579 A 7, 1990 Vishnuvajala et al...... 514,283 5,635,611 A 6/1997 Ishiguro et al...... 536, 4.1 geniposide, rubusoside, rebaudioside A, Steviol monoside 6,673,843 B2 1/2004 Arbiser ...... 514f679 and stevioside) were discovered to enhance the solubility of a 2002/0076426 A1 6/2002 Zirnstein et al. ... 424/401 number of pharmaceutically and medicinally important com 2006,0003053 A1 1/2006 Ekanayake etal ... 426,51 pounds, including but not limited to, paclitaxel, camptoth 2007, OO32438 A1 2/2007 Hu et al...... 514/23 ecin, curcumin, tanshinone HA, capsaicin, cyclosporine, 2008/0242691 A1 10, 2008 Nakazawa et al. ... 514,283 erythromycin, nystatin, itraconazole, celecoxib, clofazimine, 2011/0195161 A1* 8/2011 Upreti et al...... 426,103 digoxin, oleandrin, nifedipine, and amiodarone. The use of FOREIGN PATENT DOCUMENTS the diterpene glycoside rubusoside and monoterpene glyco JP 58-101660 6, 1983 side paenoiflorin increased solubility in all tested com JP O1-299235 4f1989 pounds. The terpene glycosides are a naturally occurring JP 2001-048727 2, 2001 class of water Solubility-enhancing compounds that are non JP 2001.048727 A * 2, 2001 toxic and that will be useful as new complexing agents or WO WOf 2009 126950 10/2009 excipients in the pharmaceutical, agricultural (e.g., Solubiliz OTHER PUBLICATIONS ing pesticides), cosmetic and food industries. Takashi Tanaka, Maki Kataoka, Nagisa Tsuboi, and Isao Kouno. New Monoterpene Glycoside Esters and Phenolic Constituents of 40 Claims, 21 Drawing Sheets U.S. Patent Oct. 8, 2013 Sheet 1 of 21 US 8,551,507 B2

OCH 3 Curcumin 368 Tanshinone IIA294 Curcuminoid phenol OH Quinone O

CO-Q10863 Quinone

OH O ( O 0\ ^ O H OH O O OH OCH Y OCH, Silybin 482 OH Artemisinin282 o HO O 0\, Sesquiterpene lactone Podophyllotoxin 414 Fic, F Fic, E 9. Fig 1D 9.

O O Oleandrin 576 O Digoxin 780 C final Cardiac glycoside OH Cardiac glycoside HO H. H G HO / O N/\oyN-9s-4 YoY/Yo Ho Yo ho i0 Y H H Fig. 1G Fig, 1H U.S. Patent Oct. 8, 2013 Sheet 2 of 21 US 8,551,507 B2

OH it. t/ ch, ( OF HOH (H6H 0 t Y Y-ch. 3.11. o: HCOONa M. I./Y. M CH Camptothecin348 O Y., iv. HC O Quinoline alkaloid HC ^ | ych Paclitaxel 853 - Ampholericin B924 OCH, Fig Taxane Diterpene Microllide H.N-OH Fig, J Fig. K

CH, OH CH, C Clofazimine 473 Riminophenazine CH; CH, Y,\ it CeleCOxib38 e F Propofol 178 NYA. Fig. 4L Go Fia. M ^^ F. CoolFig Fig.1N

NO, : OH,C-C C-OCH, OCHCHNCHCH N/ 0 Nifedipine 346 \ AmiodarOne 645 ^ Dihydropyridine CHCHCHCH. Furan H.C N: CH, O Fig, 10 Fig, 1P U.S. Patent Oct. 8, 2013 Sheet 3 of 21 US 8,551,507 B2

OH Mogroside V OAC Astragaloside IV

OH U.S. Patent Oct. 8, 2013 Sheet 4 of 21 US 8,551,507 B2

HO CHOn 20 O E O

O

HO CHOH O O 0 H OH O OH GenipOside Paeoliflorin OH Fig. 2C Fig. 2D U.S. Patent Oct. 8, 2013 Sheet 5 of 21 US 8,551,507 B2

H

HO O OH

HO O O OH

O Stevioside

Fig. 3A Fig. 3B U.S. Patent Oct. 8, 2013 Sheet 6 of 21 US 8,551,507 B2

HO OH O HO OH OH

O

OH

Rebaudioside -0 SteviolmOnOside HO OH Fig. 3C Fig. 3D U.S. Patent Oct. 8, 2013 Sheet 7 of 21 US 8,551,507 B2

Curcumin O36

O27

O.O. wCUR6 CUR5 CUR4 CUR3 CUR CUR 20 280 50 420 490 B6 30 700 to 840 Minutes Fig. 4 U.S. Patent Oct. 8, 2013 Sheet 8 of 21 US 8,551,507 B2

3.

8 “renewauw“wswww.s.l...rew-ways errorway.' Ney'." . , '... w www.'i '"raw"westwytoves g; 23: 48 BCC 8 OCC 200 BC 5). it: 8 Fig.5 U.S. Patent Oct. 8, 2013 Sheet 9 of 21 US 8,551,507 B2

Paclitaxel 100 to

750 of

3. t

2 : 3. 100. 1200 - 0. 600 800 2008 Miyas Fig 6 U.S. Patent Oct. 8, 2013 Sheet 10 of 21 US 8,551,507 B2

O50 0.48 O46 O44 O42 0.40 O38 0.36 O34 O32 O3O O28 O26 ; : 024 Tanshinone IIA g O22 : O20 0.18 0.16 O.14 O. 12 0.10 0.08 0.06 004 0.02". OOOo -002 -004 OOO 2.OO 4OO 6OO 800 OOO 1200 14 OO 16 OO 18OO 20 OO 22 OO 20 260 28 OO 3OOO 32OO 3400 Minutes Fig. 7 U.S. Patent Oct. 8, 2013 Sheet 11 of 21 US 8,551,507 B2

Amphotericin B

Minutes Fig. 8 U.S. Patent Oct. 8, 2013 Sheet 12 of 21 US 8,551,507 B2

to Artemisinin

-OOO OOO 200 400 800 800 OOO 12 OO 1400 1600 1800 20 OO 2200 24 OO 26 OO 2800 3OOO 3200 3400 Minutes Fig, 9 U.S. Patent Oct. 8, 2013 Sheet 13 of 21 US 8,551,507 B2

Podophytotoxin

:

20: ...... E is 18 to 22, 20 30 3. Rigtes Fig. 10 U.S. Patent Oct. 8, 2013 Sheet 14 of 21 US 8,551,507 B2

O2OO O. 190 0.18O O 170 0.160 O150 0,140 0.130 O. 120 OO OOO o O080 OOTO OO60 0.050 0.040 0.030 OO20 OOO OOOO. -OOO -0,020 OOO 2.00 400 600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 3000 3200 34.00 Minutes Fig. 1 U.S. Patent Oct. 8, 2013 Sheet 15 of 21 US 8,551,507 B2

400 380 360 340 320 300 280 : 260 Propofol

220 s 2OO 180 Bo 140 120 1OO O80 O60 O40" 020." OOOT

OOO 2 OO 4OO 600 8 OO 10 OO 12 OO 1400 1600 18 OO 2000 22 OO 2400 26 OO 28 OO 3OOO 32OO 3400 Minutes Fig 12 U.S. Patent Oct. 8, 2013 Sheet 16 of 21 US 8,551,507 B2

2 . Soo 80 to go do Goo Go 2.202 Boo 2:03. it tes Fig. 13 U.S. Patent Oct. 8, 2013 Sheet 17 of 21 US 8,551,507 B2

28O Clofazimine 260

240

220 2OO 18O

16O 140 --

OOO o.O 20 406080 1 0 12040 160 18.0 200202402028.000320340 Minutes Fig. 14 U.S. Patent Oct. 8, 2013 Sheet 18 of 21 US 8,551,507 B2

8

. I I : I ; ; : I O. : E SO 80 6 OG 8, 2 2GO 2: OG 26, 28 O 3G sites Fig. 15 U.S. Patent Oct. 8, 2013 Sheet 19 of 21 US 8,551,507 B2

ites Fig, 16 U.S. Patent Oct. 8, 2013 Sheet 20 of 21 US 8,551,507 B2

s

O 2. a 4G so 8 to 20 460 to 2002. 2. 80808 (3.08.0 Rites Fig. 17 U.S. Patent Oct. 8, 2013 Sheet 21 of 21 US 8,551,507 B2

a

s s 8. 0. l i also go on 22 on 2 is a 323i inites Fig 18 US 8,551,507 B2 1. 2 TERPENE GLYCOSDES AND THEIR Micelles can vary in sizes, some of which are nanosized, COMBINATIONS AS SOLUBLIZINGAGENTS referred as nanomicelles. Surfactants are classified based on the charge of its hydrophilic groups: ionic (e.g., anionic, This is the United States national stage of international cationic) and non-ionic Surfactants. Surfactants have many application PCT/US2010/039799, international filing date applications, for example, detergents, emulsifiers, cosmetics, Jun. 24, 2010, which claims the benefit of the Jun. 24, 2009 and solubilizers. Some of the surfactants that are used for filing date of U.S. provisional patent application 61/219,973 solubilizing water-insoluble drugs include propylene glycol, under 35 U.S.C. S119(e). glycerol, Sodium lauryl Sulfate, and phospholipids. Micelles have many advantages in pharmaceutical formulation but TECHNICAL FIELD 10 solubilizing efficiency has been one of the major obstacles. New effective surfactants that are non-toxic natural ingredi This invention pertains to new compositions and uses for ents and superior to the currently available surfactants hold certain terpene glycosides as non-toxic, natural Solubilizers promises of overcoming the limitations and advancing the for use in preparing aqueous solutions of various drugs, agri formulations of pharmaceutical, cosmetic, agricultural cultural chemicals, cosmetics, and foods. 15 chemicals, and foods products. Important Compounds Insoluble in Water BACKGROUND ART Diterpenes. Taxanes are diterpenes produced by the plants of the genus Drug Solubility and Complexing Agents Taxus (yews) such as the Pacific Yew (Taxus brevifolia) in the family of Taxaceae. Taxanes include paclitaxel and docetaxel. Poor aqueous solubility is a common obstacle to delivering Paclitaxel is the anti-cancer drug under the drug name of pharmaceuticals or other bioactive compounds and is a major TAXOL(R) and docetaxel is used under the name of TAXO challenge in formulating new drug products. In a study of TERER (Medicinal Natural Products A Biosynthetic kinetic aqueous solubility of commercial drugs, 87% were Approach, 1997, John Wiley & Sons, Chichester, England; pp found to have solubility in water of >65 lug/mL and 7%s20 25 186-188) Paclitaxel is a known anti-cancer diterpenoid alka ug/mL (Lipinski, C., et al., Adv. Drug Deliv. Rev. (1997) loid and is not soluble in water. The structure of paclitaxel is 23:3-25). The minimum acceptable aqueous solubility for a shown in FIG. 1.J. Therapeutic solutions of paclitaxel cur drug is about 52 g/mL Solubility based on 1 mg/kg clinical rently contain either oil or dehydrated alcohol or both; or dose and average permeability (C. A. Lipinski, J Pharm Tox paclitaxel is bound to albumin. None of these formulations Meth (2000) 44:235-249). The pharmaceutical industry has 30 are true water solutions. Other taxanes include baccatin III, been employing various approaches to increasing water-in 10-deacetylbaccatin III, cephalomannine, and 10-deacetyl soluble drugs for pharmaceutical drug formulations. Com cephalomannine. These taxanes are characterized with a four monly used approaches are the uses of one or more complex membered oxetane ring and a complex ester side-chain in ing agents (e.g., cyclodextrins), cosolvents (e.g., ethanol, their structures. All taxane compounds have poor water solu polyethylene glycol), Surfactants (e.g., Cremophor EL, 35 bility. (U.S. Patent Application Publication no. 2007/ Tween 80), emulsifiers (e.g., lecithin, glycerol), and lipo 0032438). Other medicinally important, but insoluble or Some, micelle and nanosuspension techniques, alone or in poorly soluble diterpenes include retinoids (vitamin A, ret combinations. Within this group, the use of complexing inol (vitamin A1), dehydroretinol (vitamin A2), retinoic acid, agents to improve solubility of water-insoluble drugs is 13-cis-retinoic acid and other retinol derivatives, ginkgolides increasing. Complexing agents improve water Solubility by 40 and forsakolin (a promising drug for the treatment of glau forming a non-covalent stoichiometric association with the coma, congestive heart failure, and bronchial asthma). pharmaceutical drug. Currently, the main complexing agents Quinoline Alkaloids. in the pharmaceutical industry are various forms of cyclodex Quinoline alkaloids are alkaloids that possess quinoline in trins (“CDs, molecular weight around 1135 Daltons), which their structures and are terpenoid indole alkaloid modifica form inclusion complexes with water-insoluble drug. The use 45 tions. Camptothecins isolated from the Camptotheca acumi of cyclodextrin inclusion complexation has successfully nata trees (Family Nyssaceae) are quinoline alkaloids. Camp solubilized many insoluble drugs, including an antifungal, tothecin (CPT) is a cytotoxic alkaloid and is reported to have Voriconazole, and an antipsychotic, Ziprasidone mesylate, anti-tumor properties, perhaps by inhibiting topoisomerase 1. which use sulfobutylether-f-cyclodextrin as the complexing (See, for example, U.S. Pat. No. 4,943,579). The structure of agent. The most important cyclodextrins are parent C.-, 3-, and 50 camptothecin is shown in FIG. 1I. It has poor solubility in Y-CD as well as two modified hydroxypropyl-3-CD and sul water (The Merck Index, 1996). Semi-synthetic analogues of fobutylether-B-CD. However, even the use of cyclodextrins camptothecins such as topotecan and irinotecan are approved has its disadvantages. Some of these limitations include lack chemotherapeutic drugs. Natural camptothecins include of compatibility of the drug molecules with the inclusion camptothecin, 10-hydroxycamptothecin, methoxycamptoth cavity of CDs, precipitation of the formed complexes of CD 55 ecin, and 9-nitrocamptothecin. None of the natural camptoth drug during dilution (e.g., in the stomach), potential toxicity ecins are water soluble (see, for example, U.S. Patent Appli and quality control of uniform CDs, and low complexation cation Publication no. 2008/0242691). Camptothecins have efficiency for achieving desirable solubility effect. Therefore, broad-spectrum anti-cancer activity, but poor water Solubility new complexing agents that are Superior to cyclodextrins in has limited direct uses as chemotherapeutic agents. Other overcoming or reducing these limitations are needed for the 60 quinoline alkaloids include the long recognized anti-malarial formulations of pharmaceutical, cosmetic, agricultural drugs quinine, quinidine, cinchonidine, and cinchonine. chemicals, and foods products. Surfactant compounds are Curcuminoids/Phenols. widely used in solubilizing pharmaceutical compounds Curcuminoids/phenols are a class of compounds found in because Surfactants possess amphiphilic property and can turmeric spice from the plant, Curcuna longa, of the ginger form micelles or liposomes in water. The water-soluble 65 family. Curcuminoids include, for example, curcumin, des micelles or liposomes have hydrophobic cores that can host methoxycurcumin, and bis-desmethoxycurcumin. Other phe water-insoluble molecules thus improve the solubility. nols include, for example, tocopherol (vitamin E), propofol. US 8,551,507 B2 3 4 and gingerols. Curcumin is an orange-yellow pigment that is ces ambofaciens, tylosin (16-membered) from Streptomyces found in the rhizome of Curcuma longa, the Source of the fradiae, and avermectins (16-membered with a long spice turmeric. The structure of curcumin is shown in FIG. polyketide chain). Some examples of polyene macrollides are 1A. Curcumin has been reported to have several beneficial amphotericin B from Streptomyces nodosus, nystatin from properties, including promotion of general health, anti-in 5 Streptomyces noursei, tacrolimus (23-membered) from flammatory and antimicrobial properties, and treatment for Streptomyces tsukubaensis, and rapamycin (sirolimus; digestive disorders. (See, for example, U.S. Pat. No. 6,673, 31-membered). 843) Curcumin is a lipophilic compound that is insoluble in Amphotericin B is a polyene antifungal, antibiotic from water (The Merck Index, 1996). Alpha-tocopherol, one of the Streptomyces and has antimicrobial spectrum covering yeast most potent forms of Vitamin E, is a lipid-soluble phenol 10 and other fungi. It is a yellowish powder that is insoluble in compound that is not soluble in water. Gingerols are lipid water. The structure of amphotericin B is shown in FIG. 1K. soluble phenol compounds primarily isolated from the root of Examples of applications of Amphotericin B: (1) antifungal: ginger (Zingiber officinale). Gingerols (e.g., 6-gingerol) may use of oral liposomal preparations of Amphotericin B to treat reduce nausea caused by motion sickness or pregnancy and fungal disease, e.g., thrush; (2) use in tissue culture to prevent may also relieve migraine. 15 fungi from contaminating cell cultures. It is usually sold in a Propofol is a drug for anesthetic and hypnotic uses. Cur concentrated lipid complex/liposomal solution, either on its rently, there are two drug forms using propofol. Its structure is own or in combination with the antibiotics penicillin and shown in FIG.1L. Propofol is formulated as an emulsion of a streptomycin; (3) use as an antiprotozoal drug in otherwise soybean oil/propofol mixture in water. Newer generic formu untreatable parasitic protozoan infections such as visceral lations contain sodium metabisulfite or benzyl alcohol. Pro leishmaniasis and primary amoebic meningoencephalitis; pofol emulsion (also known as “milk of amnesia') is a highly and (4) use as an antibiotic in febrile, immunocompromised opaque white fluid. The drug is sold as 200 mg propofol in 20 patients who do not respond to broad-spectrum antibiotics. mL emulsifier (1%). The other drug form of propofol is a An aqueous formulation of amphotericin would offer new water-soluble form of the drug, fospropofol. ways to administer this important drug, including intravenous Quinones. 25 SC. Quinones are a class of compounds having a fully conju Sesquiterpene Lactones. gated cyclic dione structure. This class includes, for example, Sesquiterpene lactones are a class of sesquiterpenes (15 ubiquinones (coenzyme Q. Such as coenzyme Q10), plasto carbon compounds) containing a lactone. Examples of quinones, (e.g., rhein, emodin, alizarin, and insoluble sesquiterpenes are artemisinin (a new, highly-effec lucidin), phenanthraquinones (e.g., cryptotanshinone, tanshi 30 tive anti-malarial compound), dihydroartemissinin, and bilo none I, tanshinone IIA, and dihydrotanshinone), and di-an balide (isolated from Ginkgo biloba). thraquinones (e.g., sennosides A and B). For example, tan Artemisinin is a sesquiterpene lactone drug used to treat shinone IIA is one of the natural analogues oftanshinone. The multi-drug resistant strains of falciparum malaria. Artemisi structure oftanshinone IIA is shown in FIG. 1B. Tanshinones nin is isolated from the plant Artemisia annua, but can also be have been reported to have various physiological activities 35 synthesized from artemisinic acid. Its structure is shown in from attenuating hypertrophy incardiac myocytes to aiding in FIG. 1D. Artemisinin is poorly soluble, which limits its bio treatment of obesity. (See, for example, U.S. Patent Applica availability. Semi-synthetic derivatives of artemisinin, tion Publication 2007/0248698). Tanshinone IIA (as well as including artemether and artesunate, have been developed. other tanshinones such as tanshinone I) is soluble in methanol However, their activity is not long-lasting, with significant but insoluble in water. 40 decreases in effectiveness after one to two hours. To counter Another quinone is coenzyme Q10 (often abbreviated as this drawback, artemisinin is given with lumefantrine (also CoQ10), a benzoquinone. The structure of CoQ10 is shown in known as benflumetol) to treat uncomplicated falciparum FIG. 1C. This oil-soluble vitamin-like substance is a compo malaria. Lumefantrine has a half-life of about 3 to 6 days. nent of an electron transport chain in aerobic cellular respi Such a treatment is called ACT (artemisinin-based combina ration. CoO10 acts as an antioxidant and is often used as a 45 tion therapy); other examples are artemether-lumefantrine, dietary supplement. The problems with CoQ10 are its insolu artesunate-mefloquine, artesunate-amodiaquine, and artesu bility in water and low bioavailability. Several formulations nate-Sulfadoxine/pyrimethamine. Recent trials have shown have been developed and tested on animals or humans includ that ACT is more than 90% effective, with recovery from ing attempts to reduce the particle size and increase Surface malaria after three days, even with chloroquine-resistant area of the compound, soft-gel capsules with CoQ10 in oil 50 Plasmodium falciparum. A water Solution of artemisinin Suspension, the use of aqueous dispersion of Solid CoQ10 would be highly desirable for direct parenteral applications. with tyloxapol polymer, formulations based on various Solu . bilizing agents, i.e. hydrogenated lecithin, and complexation Lignans are a class of compounds in which two phenylpro with cyclodextrins, carriers like liposomes, nanoparticles, pane monomer units are coupled at the and dendrimers. Solubilizing CoQ10 in a water solution 55 central carbon of the side-chain (lignans) or at another loca could have many uses as new medical treatments, including tion (neolignans). Examples of lignans are podophyllotoxin the administration by injection. (isolated from American Mayapple), 4'-demethylpodophyl Microllides. lotoxin, beta-peltatin, alpha-peltatin, desoxypodophyllo Microllides are a large family of compounds, many with toxin, podophyllotoxone, , yatein, and pinores antibiotic activity, characterized by a macrocyclic lactone 60 inol. Podophyllotoxin, also known as codylox or podofilox, is ring typically 12-, 14-, or 16-membered (reflecting the num a lignan compound, and a non-alkaloid toxin isolated from ber of units used), but can also be even larger polyene mac the rhizome of American Mayapple (Podophyllum peltatum). rolides with microllide ring size ranging from 26 to 38-mem Its structure is shown in FIG. 1E. Podophyllotoxin can also be bered. Some examples of typical macrollides are synthesized biologically from two molecules of coniferyl erythromycins (14-membered) from Streptomyces erythreus, 65 alcohol. Podophyllotoxin is the pharmacological precursor oleandomycin (14-membered) from Streptomyces antibioti for the important anti-cancer drug etoposide. It is also admin cus, spiramycin I, II, and III (16-membered) from Streptomy istered to treat genital warts. Podophyllotoxin is poorly US 8,551,507 B2 5 6 soluble in water, and a water solution containing a pharma Dihydropyridine. ceutically effective amount has not been available. Dihydropyridines is a class of drugs as calcium channel . blockers. Examples of dihydropyridine include nifedipine, amlodipine, aranidipine, azelnidipine, barnidipine, benid Flavonolignans are a class of compounds structurally com ipine, cilnidipine, clevidipine, efonidipine, felodipine, israd bined from and lignan. These include compounds ipine, lacidipine, manidipine, lercanidipine, nicardipine, nil Such as Silybin, isosilybin, and sillychristin (seen in the plant Vadipine, nimodipine, nisoldipine, nitrendipine, and of milk thistle ( marianum) from the family of Com pranidipine. The structure of nifedipine is shown in FIG. 10. positae. Silybin, also known as Silibinin, is the major active Nifedipine is a calcium channel blocker drug, currently for constituent of silymarin, the mixture of flavonolignans mulated in capsules. No injectable drugs are currently avail extracted from milk thistle (Silybum marianum). The struc 10 able. ture of silybin is shown in FIG. 1F. Studies suggest that Amiodarone. Silybin has hepatoprotective (antihepatotoxic) properties and Amiodarone is Class III agent (a potassium channel anti-cancer effects against human prostate adenocarcinoma blocker) and an antiarrhythmic agent (medication used for cells, estrogen-dependent and estrogen-independent human irregular heartbeat) used for various types of tachyarrhyth 15 mias (fast forms of irregular heartbeat), both ventricular and breast carcinoma cells, human ectocervical carcinoma cells, Supraventricular (atrial) arrhythmias. The structure of amio human colon cancer cells, and both Small and nonsmall darone is shown in FIG. 1 P. Amiodarone is an antianginal and human lung carcinoma cells. Poor water Solubility and bio antiarrhythmic drug, currently available in oral and injection availability of silymarin led to the development of enhanced formulations. formulations. Silipide (trade name SILIPHOSR), a complex Riminophenazine. of silymarin and (lecithin), is about ten Riminophenazine is a class of fat soluble dyes used for the times more bioavailable than silymarin. It has been also treatment of leprosy. It has been used investigationally in reported that silymarin inclusion complex with B-cyclodex combination with other anti-mycobacterial drugs to treat trin is much more soluble than silymarin itself. Glycosides of Mycobacterium avium infections in AIDS patients. One silybin show better water solubility and even stronger hepato 25 example is clofazimine that has a marked anti-inflammatory protective effects. However, an aqueous solution of silybin in effect and is given to control the leprosy reaction, erythema pharmaceutically acceptable amount, in its original and nodosum leprosum. (From AMA Drug Evaluations Annual, unmodified structure, has not been available for parenteral 1993, p 1619). The structure of clofaximine is shown in FIG. administrations. 1N. Clofazimine is a non-steriodal, anti-inflammatory drug Azole. 30 and is currently formulated using propylene glycol, oils, and An azole is a class of five-membered nitrogen heterocyclic beeswax. ring compounds containing at least one other noncarbon Terpene Glycosides atom, for example, a nitrogen, Sulfur or oxygen (Eicher, T.; Natural terpene glycosides are well known and exist in a Hauptmann, S. (2nd ed. 2003). The Chemistry of Hetero variety of plant Sources. They generally are terpeneaglycons cycles: Structure, Reactions, Syntheses, and Applications. 35 attached to at least one glucose, and the most common forms Wiley-VCH. ISBN 3527307206). Itraconazole is a triazole are diterpene glycosides and triterpene glycosides. Many of with antifungal activities. Other triazole antifungal drugs these compounds are known to be non-toxic and natural include fluconazole, isavuconazole, Voriconazole, prami sweeteners. (U.S. Published Patent Application No. 2006/ conazole, posaconazole, ravuconazole, fluconazole, fosflu 000305053). Rubusoside is a diterpene glycoside that is from conazole, epoxiconazole, triadimenol, propiconazole, met 40 Chinese Sweet leaf tea leaves (Rubus suavissimus; Rosaceae). conazole, cyproconazole, tebuconazole, flusilaZole and Rubusoside has a molecular formula C32H50013 and paclobutraZol. These compounds are practically insoluble in molecular weight of 642.73. (From T. Tanaka et al., Rubuso water (e.g., itraconazole, The Merck Index, 1996, p. 895). side (b-D-glucosyl ester of 13-O-b-D-glucosyl-steviol), a Itraconazole has relatively low bioavailability after oral Sweet principle of Rubus chingii Hu (Rosacease), Agricul administration. 45 tural and Biological Chemistry, Vol. 45(9), pp. 2165-6, 1981). Celecoxib is a pyrazole (a rare alkaloid), a compound that Rubusoside also has good solubility in water, alcohol and targets cyclooxygenase (COX) enzymes. The structure of acetone ethyl acetate. It is a diterpene aglycone with two celecoxib is shown in FIG. 1M. In medicine, pyrazoles are glucose molecules attached. (FIG. 3A) Diterpene glycosides used for their analgesic, anti-inflammatory, antipyretic, anti have been shown by me to be good natural solubilizers. (Inter arrhythmic, tranquilizing, muscle relaxing, psychoanaleptic, 50 national application no. PCT/US2008/040324; international anticonvulsant, monoamineoxidase inhibiting, antidiabetic publication no. WO 2009/126950). The entire contents of that and antibacterial activities. Celecoxib is a COX-2 inhibitor. published application is fully incorporated into this applica Celecoxib has poor solubility in water which reduces its tion. Another diterpene glycoside that is isolated from the bioavailability. Chinese Sweet leaf tea (Rubus suavissimus; Rosaceae) is Ste Cardiac Glycosides. 55 viol monoside (FIG. 3D). The structure of steviol monoside Cardiac glycosides are drugs used in the treatment of con has only one glucose molecule rather than two as in rubuso gestive heart failure and cardiac arrhythmia. The therapeutic side. Steviol monoside can be isolated from the sweet leaf tea functions, however, depend on the structure of the steroidal or be obtained through the acid hydrolysis of rubusoside to aglycone and the type and number of Sugars attached. Based cleave one glucose molecules. Unlike rubusoside, Steviol on the structure of aglycone, two types of cardiac glycosides 60 monoside is not a dominant diterpene glycoside in the Sweet are named: cardenolides (e.g., digoxin, oleandrin) and bufa leaf tea plant. dienolides (e.g., hellebrigenin). The structure of digoxin is Stevioside is a diterpene (steviol)glycoside that is isolated shown in FIG. 1G. Digoxin is used to treat congestive heart from the Stevia leaf (Stevia rebaudiana; Asteraceae). Stevio failure; the drug is formulated as an injection Solution con side has a molecular formula CHOs and a molecular taining 40% propylene glycol and 10% alcohol to achieve 250 65 weight of 804. The compound as shown in FIG. 3B is a ug/mL concentration. Oleandrin may be used to treat conges diterpene aglycone with three glucose molecules. In pure tive heart failure or cancer. form, it is a crystal or white powder. Another diterpene gly US 8,551,507 B2 7 8 coside that is isolated from the Stevia leaf is rebaudioside A. bin (FIG.1F), digoxin (FIG.1G), oleandrin (FIG. 1H), camp The compound as shown in FIG. 3C is a diterpene aglycone tothecin (FIG. 1I), paclitaxel (FIG. 1J), amphotencin (FIG. with four glucose molecules. In pure form, it is a white pow 1K), propofol (FIG. 1L), celecoxib (FIG. 1M), clofazimine der. (FIG. 1N), nifedipine (FIG. 10), and amiodarone (FIG. 1P). Mogroside V is a triterpene glycoside that is isolated from FIGS. 2A-2D illustrate the structure of some representative the luohanguo fruit (Siraitia grosvenorii, formerly known as triterpene glycosides (mogroside V. FIG. 2A; and astragalo Momordica grosvenori; Curcubitaceae). The structure of side, FIG. 2B) and monoterpene glycosides (paconiflorin, mogroside V is shown in FIG. 2A. The Sweet taste of luo FIG. 2C; and geniposide, FIG. 2D). hanguo comes mainly from the mogrosides, a group of trit FIGS. 3A-3D illustrate the structure of representative erpene glycosides that makeup approximately 1% of the flesh 10 diterpene glycosides, including rubusoside (FIG.3A), Stevio of the fresh fruit. Five different mogrosides are known, named side (FIG. 3D), rebaudioside A (FIG. 3C), and steviol mono with the numbers 1 to 5. The main mogroside in luohanguo is side (FIG. 3D). mogroside V. Mogroside V has a molecular formula FIG. 4 illustrates the results of high performance liquid CoHoO and a molecular weight of 1286. A second trit chromatography indicating the amount of dissolved cur erpene glycoside is astragaloside IV with a structure as shown 15 cumin in six solutions: CUR1, 10% mogroside V: CUR2, in FIG.2B. One triterpene glycoside was reported to increase 10% astragaloside: CUR5, 10% geniposide; CUR4, 10% the solubility of saikpsaponin A. See H. Kimata et al., Chem. paconiflorin: CUR5, 10% rubusoside; and CUR6, 10% ste Pharm. Bull. (Tokyo), vol. 33:2849-2853, 1985; andY. Sasaki viol monoside. et al., Chem. Pharm. Bull. (Tokyo), vol. 36:3491-3495 FIG. 5 illustrates the results of high performance liquid (1988). chromatography indicating the amount of dissolved camp Monoglycosides are also known. Two monoglycosides are tothecin in five solutions: CPT1, 10% mogroside V: CPT3, paenoiflorin and geniposide. The structures of these two com 10% geniposide; CPT4, 10% paconiflorin: CPT8, 10% rebau pounds are shown in FIGS. 2C and 2D, respectively. Paeoni dioside A; and CPT9, water (control). florin has been reported to increase the solubility of poly FIG. 6 illustrates the results of high performance liquid meric proanthocyanidins. Both were found in the extract from 25 chromatography indicating the amount of dissolved pacli Paeonine Radix (Shaoyao), an important drug in Japanese taxel in five solutions: TXL1, 10% mogroside V: TXL3, 10% and Chinese traditional medicine. See, for example, T. Tanaka geniposide; TXL4, 10% paeoniflorin; TXL8, 10% rebaudio et al., Chem. Pharm. Bull., vol. 48(2): 201-207, 2000; and T. side A; and TXL9, water (control). Tanaka et al., Chem. Pharm. Bull., vol. 45(12): 1891-1897 FIG. 7 illustrates the results of high performance liquid (1997). 30 chromatography indicating the amount of dissolved tanshi U.S. Published Patent Application No. 2002/0076426 dis none IIA in six solutions: Tan-1, 10% geniposide; Tan-2, 10% closes terpene alcohol ethoxylates as solubilizers in pharma paeoniflorin; Tan-3, 10% mogroside V: Tan-4, 10% astraga ceutical and food preparations. loside IV: Tan-5, water (control); and Ref, 30.4 ug/ml tanshi Chinese Patent No. 1723981 discloses that an extract con none IIA in methanol (reference standard). taining triterpene glycosides (mogrosides) isolated from 35 FIG. 8 illustrates the results of high performance liquid Momordica grosvenoiri fruit was used to replace Sucrose or chromatography indicating the amount of dissolved ampho other Sweeteners in manufacturing pills, granules, tablets, tericin B in six solutions: Amp-1, 10% geniposide; Amp-2, capsules or Solutions of traditional Chinese medicine. 10% paeoniflorin; Amp-3, 10% mogroside V: Amp-4, 10% astragaloside IV: Amp-5, water (control); and Ref, 55ug/ml DISCLOSURE OF INVENTION 40 amphotericin B in methanol (reference standard). FIG. 9 illustrates the results of high performance liquid I have discovered that mogroside V, paeoniflorin, and geni chromatography indicating the amount of dissolved artemisi poside enhanced the solubility of a number of pharmaceuti nin in six solutions: Art-1, 10% geniposide; Art-2, 10% pae cally and medicinally important compounds of several struc oniflorin; Art-3, 10% mogroside V: Art-4, 10% astragaloside tural classes, including but not limited to, the important 45 IV: Art-5, water (control); and Ref 730 g/ml artemisinin in water-insoluble drugs of paclitaxel, camptothecin, curcumin, methanol (reference standard). tanshinone IIA, amphotericin B, artemisinin, podophyllo FIG. 10 illustrates the results of high performance liquid toxin, Silybin, propofol, celecoxib, clofazinine, digoxin, ole chromatography indicating the amount of dissolved podo andrin, nifedipine, and amiodarone. The use of the above phyllotoxin in six solutions: Pod-1, 10% geniposide: Pod-2, glycosides increased solubility of all tested compounds from 50 10% paeoniflorin: Pod-3, 10% mogroside V: Pod-4, 10% about 2-fold to over 1000-fold, depending on the compound. astragaloside IV: Pod-5, water (control); and Ref, 370 ug/ml In addition, certain diterpene glycosides (rubusoside, Steviol podophyllotoxin in methanol (reference standard). monoside, Stevioside, and rebaudioside A) were shown to FIG. 11 illustrates the results of high performance liquid enhance the solubility of clofazinine, digoxin, oleandrin, chromatography indicating the amount of dissolved Silybin in nifedipine, and amiodarone. The use of terpene glycosides as 55 six solutions: Sil-1, 10% geniposide: Sil-2, 10% paeoniflorin; naturally occurring water solubility-enhancing compounds Sil-3, 10% mogroside V: Sil-4, 10% astragaloside IV: Sil-5, for specific drugs and compounds will be useful in the phar water (control); and Ref, 26 ug/ml Silybin in methanol (ref. maceutical, agricultural, cosmetic, and food industries. erence standard). FIG. 12 illustrates the results of high performance liquid BRIEF DESCRIPTION OF DRAWINGS 60 chromatography indicating the amount of dissolved propofol in six solutions: PRO-1, 10% geniposide; PRO-2, 10% pae FIGS. 1A to 1P illustrates the structures of representative oniflorin: PRO-3, 10% mogroside V: PRO-4, 10% astragalo compounds of several classes of compounds that are knownto side IV: PRO-5, water (control); and Ref, 702 g/ml propofol have low water solubility, and that have been shown to be in methanol (reference standard). solubilized using a terpene glycoside, including curcumin 65 FIG. 13 illustrates the results of high performance liquid (FIG. 1A), tanshinone IIA (FIG. 1B), Coenzyme-Q10 (FIG. chromatography indicating the amount of dissolved cele 1C), artemisinin (FIG. 1D), podophyllotoxin (FIG. 1E), sily coxib in six solutions: Cel-1, 10% geniposide; Cel-2, 10% US 8,551,507 B2 9 10 paeoniflorin: Cel-3, 10% mogroside V: Cel-4, 10% astraga has been increased by a factor of 5 or more, in others by a loside IV: Cel-5, water (control); and Ref, 84 ug/ml celecoxib factor of 10 or more, in others by a factor of 20 or more, in in methanol (reference standard). others by a factor of 50 or more, in others by a factor of 100 or FIG. 14 illustrates the results of high performance liquid more, and in others by a factor of 1000 or more. The solubi chromatography indicating the amount of dissolved clofaz 5 lizers can be used in concentrations from 1% to 100% w/v. imine in ten solutions: C1, 10% mogroside V: C2, 10% depending on the solubility of each solubilizer in water. The astragaloside IV: C3, 10% geniposide; C4, 10% paeoniflorin; solubilzer solutions were found to be particularly effective C5, 10% rubusoside; C6, 10% steviol monoside, C7, 10%, from about 5 to about 40% w/v solubilizer, preferable about Stevioside; C8, 10% rebaudioside A: C9, water (control); and 10% w/v solubilizer. The concentration of the solubilizer will Ref. 160 ug/ml clofazimine in methanol (reference standard). 10 determine the amount of the drug that will be dissolved. Thus FIG. 15 illustrates the results of high performance liquid the concentration will depend on the desired dose of the drug chromatography indicating the amount of dissolved digoxin to be administered. in ten solutions: D1, 10% mogroside V: D2, 10% astragalo I have discovered several terpene glycosides as new solu side IV: D3, 10% geniposide; D4, 10% paeoniflorin: D5, 10% bilizing agents for creating new pharmaceutical, cosmetic, rubusoside; D6, 10% steviol monoside; D7, 10%, stevioside; 15 agricultural and food formulations instead of the commonly D8, 10% rebaudioside A: D9, water (control); and Ref, 388 used cyclodextrins. Without being bound by this theory, it is ug/ml digoxin in methanol (reference standard). believed that the improved solubility of water-insoluble drugs FIG. 16 illustrates the results of high performance liquid is a result of the formation of terpene glycoside (TGS)-drug chromatography indicating the amount of dissolved olean complex structures, such as nano- to micro-size micelles, drin in ten solutions: O1, 10% mogroside V: O2, 10% astraga which are water soluble. The driving forces for the formation loside IV: O3, 10% geniposide; O4, 10% paeoniflorin; O5, of the dTG-drug complexes may include London dispersion 10% rubusoside; O6, 10% steviol monoside; O7, 10%, Stevio forces (an induced dipole-induced dipole attraction), dipolar side; O8, 10% rebaudioside A, O9, water (control); and Ref. forces (including hydrogen-bonding), ionic (electrostatic) 260 g/ml oleandrin in methanol (reference standard). forces, and/or hydrophobic effects as described in R. Liu, FIG. 17 illustrates the results of high performance liquid 25 Water-insoluble drug formulation, Second Edition, pp 133 chromatography indicating the amount of dissolved nifide 160, 2008, CRC Press, Boca Raton, Fla. Depending on the pine inten solutions: N1, 10% mogroside V: N2, 10% astraga drug molecule, solubilization power of the TGs will vary loside IV: N3, 10% geniposide: N4, 10% paeoniflorin; N5, depending on the driving force in forming each intermolecu 10% rubusoside; N6, 10% steviol monoside; N7, 10%, Stevio lar complexation. side; N8, 10% rebaudioside A: N9, water (control); and Ref. 30 Without being bound by this theory, it is believed that the 240 g/ml nifidepine in methanol (reference standard). formation of the TG-drug complexes in aqueous solutions FIG. 18 illustrates the results of high performance liquid may be driven by similar forces proposed for cyclodextrins chromatography indicating the amount of dissolved amio (CDs) in the formation of inclusion complexes, by the sur darone in ten solutions: A1, 10% mogroside V: A2, 10% factant property, or by both. In addition to the driving forces astragaloside IV: A3, 10% geniposide; A4, 10% paeoniflorin; 35 above, van der Waals forces (the attractive or repulsive force A5, 10% rubusoside; A6, 10% steviol monoside; A7, 10%, between molecules or between parts of the same molecule) Stevioside; A8, 10% rebaudioside A: A9, water (control); and may be involved. The difference between the CD-drug inclu Ref. 104 ug/ml amiodarone in methanol (reference standard). sion complexes and the TG-drug complexes may be attribut able to their geometrical structures. Rather than forming a MODES FOR CARRYING OUT THE INVENTION 40 circle with a hydrophobic cavity similar to the CDs, the TGs may form uniform and dynamic structures, with the hydro Several important organic compounds are insoluble in philic glucose molecules exposing to water and with the water or have very low solubility. I have tested many of these hydrophobic terpene aglycones in the center as the spacer therapeutic compounds from several classes of chemical sites that host water-insoluble drug molecules. structures and found that natural solubilizers based onterpene 45 The new complexing or micellating agents, terpene glyco glycosides have increased the aqueous solubility of many sides (TGS), have several advantages over CDS as complexing compounds tested. I have found a method for enhancing the agents. First, TGS may be less rigid on the requirement of the solubility of an organic compound which is insoluble or spar cavity size, which has been a limiting factor for the formation ingly soluble in water, said method comprising mixing said off3-CDs-drug complexes, especially large molecular drugs. compound with water and with a terpene glycoside in a con 50 Second, the possible uniformity of hydrophilic-hydrophobic centration sufficient to increase the solubility of the com spacing alignment of TGS may be more efficient than the pound in water by a factor of 2 or more. The solubility for the circular hydrophilic-hydrophobic spacing alignment, and organic compounds in some cases has been increased by a thus capable of solubilizing more drug molecules. Third, the factor of 5 or more, in others by a factor of 10 or more, in TGs have excellent water solubility and stability in water others by a factor of 20 or more, in others by a factor of 50 or 55 solution. The solubility of some TGs can be as high as about more, in others by a factor of 100 or more, and in others by a 60 g/100 mL in water at 25° C. and about 80 g/100 mL in factor of 1000 or more. water at 37°C. This is much higher than B-CD of 1.85 g/100 In addition, a new composition has been discovered com mL water, C-CD of 15 g/100 mL water, or Y-CD of 23 g/100 prising an aqueous solution of an organic compound having mL water. In addition, many of the terpene glycosides may low solubility in water, and a terpene glycoside, selected from 60 actually be safer for internal injections. Some diterpene gly the group of mogroside V, paeoniflorin, geniposide, rubuso cosides have been approved by the FDA as Sweeteners (e.g., side, Stevioside, and rebaudioside A, wherein the concentra rebaudioside A). Based on the aglycone Steviol, estimates are tion of said terpene glycoside is sufficient to increase the that daily consumption of Steviol glycosides of 8 mg rubuso solubility of said compound in water by a factor of 2 or more side/kg body weight is safe and has no adverse effect, and up above what the solubility of said compound would be in an 65 to 766 mg rubusoside/kg body weight (based on 383 mg/kg otherwise identical composition lacking said terpene glyco body weight daily expressed as Steviol) is the no-observed side. The solubility for the organic compounds in Some cases effect level. The intraperitoneal injection of stevioside water US 8,551,507 B2 11 12 solution in hypertensive rats at doses of 50 mg/kg and 100 complexes) may also be administered transdermally, for mg/kg body weight showed no adverse effects (Y.-H. Hsu et example in the form of a slow-release Subcutaneous implant, al., Antihypertensive Effect of Stevioside in Different Strains or orally in the form of capsules, powders, or granules. of Hypertensive Rats. Chinese Medical Journal (Taipei) Pharmaceutically acceptable carrier preparations for 2002; Vol. 65:1-6). In pharmaceutical dosing paradigm, 50 5 parenteral administration include sterile, aqueous or non mg/kg or less of rubusoside or other terpene glycosides may aqueous solutions, Suspensions, and emulsions. Aqueous car be sufficient to solubilize drugs to therapeutic levels for riers include water, alcoholic/aqueous solutions, emulsions parental applications. Additionally, the geometry of terpene or Suspensions, including saline and buffered media. glycosides as complexing or micellating agents to increase Parenteral vehicles include sodium chloride solution, Ring solubility of water-insoluble drugs may increase bioavailabil- 10 er's dextrose, dextrose and sodium chloride, lactated Ring ity by readily exposing the drug molecules to the bi-layer er's, or fixed oils. The solubilizer and drug may be mixed with membranes of the target cells for rapid absorption. Moreover, other excipients that are pharmaceutically acceptable and are the formed rubusoside-curcumin complexes in water Solu- compatible with the active ingredient in the drug. Suitable tions were shown resistant to heat up to 121° C. and pH excipients include water, Saline, dextrose, glycerol and etha changes from acid to alkaline conditions. Last, the heat sta- 15 nol, or combinations thereof. Intravenous vehicles include bility of certain terpene glycosides up to 250° C. allows fluid and nutrient replenishers, electrolyte replenishers, such effective use of melting and other heating methods in the as those based on Ringer's dextrose, and the like. Preserva preparation of solid complexes. Based on the above compari- tives and other additives may also be present Such as, for Sons, features, and experimental data shown in this invention, example, antimicrobials, anti-oxidants, chelating agents, it is believed that the TGs are superior to CDs as complexing 20 inert gases, and the like. agents in the solubilization of water-insoluble drugs. In addi- The form may vary depending upon the route of adminis tion, various combinations of TGs could be used as solubi- tration. For example, compositions for injection may be pro lizers to further increase the solubility of a given drug. vided in the form of an ampule, each containing a unit dose Using the terpene glycosides as solubilizers provides away amount, or in the form of a container containing multiple to alleviate problems with low solubility drugs, e.g., low 25 doses, or in the form of powder that is reconstituted with a absorption and low bio-availability of the drug. In addition, Suitable solvent (e.g., Saline solution) prior to injection. using the solubilizer and drug in a powder form (containing For purposes of this application, a compound that is solubilizer-drug complexes) will allow solid formulations insoluble in water is a compound in which less than 100 g that are readily dissolvable in water, e.g., tablet or even effer- dissolves in 1 mL water. A compound that is sparingly soluble vescent tablets. The solubilizers can be used to prepare non- 30 in water is one in which less than 20 mg, but more than 100 ug, alcoholic syrups of low solubility drugs that are stable, or to dissolves in 1 mL water. Finally, in general, a compound that prepare gelatin capsules with the solubilizer and drug inside. has low solubility in water is one in which less than 20 mg The solubilizer (terpene glycoside) and solubilized drug dissolves in 1 mL water. may be administered to a patient by any Suitable means, The structures of representative compounds of the various including oral, parenteral, Subcutaneous, intrapulmonary, 35 classes of organic compounds with low solubility are shown topical (e.g., ocular or dermal), rectal and intranasal admin- in FIGS. 1A to 1 P. A summary of some of the experimental istration. Parenteral infusions include intramuscular, intrave- data using these compounds and several natural terpene gly nous, intraarterial, or intraperitoneal administration. The cosides is given in Table 1. The details of these experiments, Solution or its dry ingredients (containing solubilizer-drug including results from control experiments, are given below. TABLE 1 Active Pharmaceutical Ingredients (APIs) in Water Solutions Containing 10% w/v Natural Solubilizing Compounds Solubility in water With With With without Mogroside With With With With Stevio With Rebaudioside Organic solubilizer V Astragaloside Geniposide Paeoniflorin Rubusoside monoside Stevioside A. Compound M.W. (g/mL) Ig/mL |g/mL |g/mL Ig/mL Ig/mL Ig/mL |g/mL Ig/mL Curcumin 348 0.6 44 O 4 5 262 4 NE NE Tanshinone IIA 294 O2 O.S O O1 23 127 NE NE NE Coenzyme Q10 863 0.1 O O O O 111 NE3 NE3 NE3 Camptothecin 348 0.4 18 NE 25 53 61 NE NE 48 Paclitaxel 853 O.35 7 NE 1 2O 65 NE NE 26 Amphotericin B 924 O2 6 O O2 19 2OO NE3 NE3 NE3 Artemisinin 282 532 148 O 146 218 28O NE NE NE Podophyllotoxin 414 92 747 O 644 S86 919 NE3 NE3 NE3 Silybin 482 O2 18 O 16 37 150 NE NE NE Propofol 178 O2 56 O 7 349 117OO NE3 NE3 NE3 Celecoxib 381 O2 47 O 3 253 488 NE3 NE3 NE3 Clofazimine 473 7.62 58 0.4 48 565 132 O 64 109 Digoxin 780 192 35 18 26 38 48 10 30 32 Oleandrin 576 32 47 5 28 42 123 7 74 67 Nifedipine 346 O2 55 O 39 49 286 O 468 2O3 Amiodarone 645 1072 238 164 235 250 249 O 2O1 271

Solubility values are obtained from MSDS, Merck Index, or publications; 'Solubility values were determined by my own analysis; NE–Not examined, US 8,551,507 B2 13 14 Example 1 Steviol monoside has only one glucose moiety (FIG. 3D) rather than two as in rubusoside (FIG. 3A). Steviol monoside Materials and Methods can be isolated from the sweet leaf tea or be obtained through the acid hydrolysis of rubusoside to cleave one glucose unit. Sources of Solubilizers Geniposide: Rubusoside: Geniposide is a monoterpene glycoside and was purchased Rubusoside was extracted from Chinese Sweet leaf tea from Shanghai University of Traditional Chinese Medicine leaves (Rubus suavissimus; Rosaceae) purchased from Natu (Shanghai, China). Its structure is shown in FIG. 2D. ral Plants Products Factory, Guilin S&T New Tech Company, Paeoniflorin: Sanlidian Campus of Guangxi Normal University, Guilin, 10 Paeoniflorin is a monoterpene glycoside and was pur Guangxi, China. Rubusoside has a molecular formula chased from Shanghai University of Traditional Chinese CHO and molecular weight of 642.73. First, the air Medicine (Shanghai, China). Its structure is shown in FIG. dried leaves were boiled with water with a weight to volume 1C. ratio ranging from about 1:10 to about 1:20. From this extrac Mogroside V. tion, a crude dried extract (20 to 30% dry weight yield from 15 Mogroside V is a triterpene glycoside that is isolated from the raw leaves) was obtained that contained from about 5% to the luohanguo fruit (Momordica grosvenori; Curcubitaceae). about 15% rubusoside by weight. The dried extract was then The structure of mogroside V is shown in FIG. 2A. A mogro reconstituted with water to a weight to Volume ratio ranging side V-rich extract (>25% by weight) was purchased from from about 1:4 to about 1:5. In this concentrated extract, the Guangxi Normal University, Guilin, China, and was used in ellagitannins would partially precipitate out and were the solubility experiments. Initial purification resulted in a removed by filtration. The rubusoside was retained in the powder containing 80% mogrosides. With further isolation Solution. The Solution containing rubusoside was then Sub and purification, a purified mogroside V (>95% purity based jected to column chromatography using a macroporous resin HPLC-UV) was obtained. This purified mogroside V was (Dowex Optipore L493 Polymeric Adsorbent, Styrene-Divi used in the solubility experiments. The purification procedure nylbenzene polymers with 46 Angstrom average pore size; 25 is described below. First, 45 g of the extract was extracted The Dow Chemical Company, Midland, Mich.). The column with 900 ml methanol with ultrasonication for 30 min twice. was eluted with ethanol to obtain a purified extract containing Then it was filtered with Whatman #4 paper filter. The filtrate approximately 60% rubusoside and about 1% steviol mono was concentrated and mixed with 25 g silica gel. The Solvent side. Some of this extract was used in Example 16 below. was removed to obtain the dried mixed sample. The dried Subsequently, the purified extract was loaded on a second 30 mixed sample was loaded onto the top of a prepared chroma column to further purify the extract using silica gel as the tography column with 600 g silica gel and a mobile phase of stationary absorbent (Silica Gel, 200-300 mesh, Natland chloroform-methanol-water (8:2:0.2). Then it was subjected International Corporation, Research Triangle, N.C.). The col to chromatography with gradient mobile phase (from water to umn was eluted with a mixed solvent (chloroform:methanol chloroform-methanol-water at 5:5:0.5 ratio). The fractions at a ratio of 8:2 V/v). The extract from this second column was 35 every 500 ml were collected. Fractions that indicated similar at least 80% pure rubusoside, and was dried to a powder. chromatograms using TLC monitoring were combined. Finally, this rubusoside-rich extract (>80% w/w) was dis Mogroside V was found in high concentration in Fraction Solved in absolute methanol by heating to temperatures rang 53-66 with light yellow color. (Data not shown) These frac ing from about 60° C. to about 80°C. The solution was then tions were combined, and 5 g active carbon was added. The cooled to allow re-crystallization of rubusoside. This re-crys 40 mixture was ultrasonicated for 30 min, and then filtered with tallization process may need to be repeated to obtain pure Whatman #4 paper filter twice. The filtrate was concentrated, rubusoside (>99% purity as measured on HPLC). The struc and Subjected to Sephadex chromatography with methanol. ture of rubusoside was confirmed by mass spectrometry and Sub-fractions were collected based on peaks monitored at NMR. Rubusoside, a diterpene glycoside, has a molecular 205 nm. The collected fractions with similar peaks were weight of 642 Daltons, and is a white crystal or powder. The 45 combined and concentrated. The concentrate was dissolved structure is shown in FIG. 3A. The crystalline powder is in 20 ml water, and the aqueous solution collected to a cen stable at temperatures ranging from about -80° C. to over trifuge tube and freeze-dried to obtain a white powder (7.59 100°C. In water, rubusoside itselfhas a solubility of approxi g). The final powder was analyzed with HPLC to confirm its mately 400 mg/ml at 25°C. and 800 mg/ml at 37°C., which purity as >95% mogroside V. MS data shows its molecularion is greater than that of many common, water-soluble com 50 (MH") is 1287.6616 (calculated 1287.6579, 2.83 ppm in pounds (e.g., Sodium chloride has a solubility of 360 mg/ml error) and molecular formula is CoHoOo. NMR data was water). collected and compared to a published reference (Data not Stevioside: shown). See Y. Xiuwei and Z. Jianye, NMR structural eluci Stevioside is a diterpene glycoside that is isolated from the dation of mogrol and its glycosides, Chinese Journal of Mag Stevia leaf (Stevia rebaudiana; Asteraceae). Stevioside has a 55 netic Resonance, 2007, 24(3): 250-260. molecular formula CHOs and a molecular weight of 804. Astragaloside IV: The structure is shown in FIG.3B. Stevioside was purchased Astragaloside is a triterpene glycoside and was purchased from Chromadex Inc. (Irvine, Calif.). from Shanghai University of Traditional Chinese Medicine Rebaudioside A: (Shanghai, China). Its structure is shown in FIG. 1B. It is not Rebaudioside A is a diterpene glycoside that is isolated 60 very soluble in water (estimated to be about 30 mg/ml in water from the Stevia leaf (Stevia rebaudiana; Asteraceae). Its or 3% w/v), and was shown below not to be an effective structure is shown in FIG.3C. Rebaudioside A was purchased solubilizer for drugs. from Chromadex Inc. (Irvine, Calif.). Compounds Tested for Solubility: Steviol Monoside: Sixteen bioactive and pharmaceutical compounds with a Steviol monoside is a diterpene glycoside that is isolated 65 water Solubility ranging from poorly soluble (200 ug/mL) to from the Chinese sweet leaf tea (Rubus suavissimus: insoluble (0.01 g/ml) were used. All compounds were found Rosaceae), the same source as rubusoside. The structure of to have purity greater than 98% based on HPLC (unless US 8,551,507 B2 15 16 otherwise indicated). Their chemical structures are shown in side (6) water Solution was each prepared. Astragaloside IV at FIGS. 1A-1P. Tanshinone IIA was purchased from Shanghai 10% w/v showed precipitation; therefore, the water solution University of Traditional Chinese Medicine (Shanghai, contained less than 10% astragaloside IV. One set of two China) and is nearly insoluble. Curcumin, camptothecin, and milligrams of curcumin was weighed into separate centrifuge paclitaxel were all purchased from Sigma-Aldrich Chemicals tubes. The solutions were sonicated at 50° C. for 60 min (St. Louis, Mo.). Artemisinin, podophyllotoxin, and Silybin (Silibinin) were purchased from LKT Laboratories (St. Paul, followed by incubation at 25°C. for 72 hr. These compounds Minn.). According to The Merck Index (1996), the solubili in the solubilized water solutions were filtered by 0.45 LM ties of these compounds areas follows: curcumin (insoluble), filters and analyzed on HPLC. Quantification was done by camptothecin (insoluble), and paclitaxel (insoluble). Accord comparing to a standard solution of a known amount in ing to the Material Safety Data Sheets, artemisinin, podo 10 methanol. The chromatograms of six curcumin water solu phyllotoxin, and Silybin (Silibinin) have a water solubility of tions (CUR1-CUR6) containing 10% w/v natural solubilizers insoluble, nearly insoluble, poor, and insoluble, respectively. (1-6: mogroside V, astragaloside, geniposide, paeoniflorin, An antifungal compound (Amphotericin B) was tested. It rubusoside, and Steviol monoside) as complexing agents, was purchased from Sigma-Aldrich Chemicals (St. Louis, respectively, are shown in FIG. 4. The HPLC system included Mo.) and is nearly insoluble. Moreover, two lipid soluble 15 600 pump, 717 autosampler, and 2996 PDA. The chromato compounds were tested: Coenzyme Q10 was purchased from grams were generated using a Prevail C18 column (4.6x250 MP Biomedicals Inc. (Solon, Ohio), and propofol were pur mm, 5um) and a mobile phase of 0.02% HCOOH-ACN (A): chased from Sigma-Aldrich Chemicals (St. Louis, Mo.). The 0.02% HCOOH-H20 (B), the gradient was A from 20% to water insoluble celecoxib was purchased from LC Laborato 80% in 45 min; at a flow rate of 1.0 mL/min, injection volume ries (Woburn, Mass.). of 5 uL., UV detection wavelength of 425 nm, and column Digoxin and clofazimine were purchased from Sigma-Al temperature of 30° C. Curcumin concentration was deter drich Chemicals (St. Louis, Mo.) and are practically insoluble mined using a standard curcumin calibration curve with cur in water (The Merck Index, 14" Edition, 2006). Oleandrin cumin standard Solutions of 2.12, 21.2, and 212 ug/ml. The was purchased from Shanghai University of Traditional Chi chromatograms of CUR1-CUR6 above were generated at 425 nese Medicine (Shanghai, China) and is practically insoluble 25 nm UV showing elution of curcumin at 39.7 min, the peak in water (The Merck Index, 14' Edition, 2006). Nifedipine areas of which were used for quantification of curcumin. The and amiodarone were purchased from LKT Laboratories (St. contents of astragaloside and Steviol monoside in Solutions Paul, Minn.) and are nearly insoluble in water. were less than 10% as judged by their precipitations in the Solubility Test Methods: water solutions. In the presence of 10% w/v of mogroside V A compound with low solubility was selected and weighed 30 (CUR1), astragaloside IV (CUR2), geniposide (CUR3), pae into multiple centrifuge tubes. Each experimental tube oniflorin (CUR4), rubusoside (CUR5), or steviol monoside received a known amount of the solubilizing agent being (CUR6), the water solutions contained 44 g/mL, 0g/mL, 4 tested. The control tubes remained only with the compound. g/mL, 5ug/mL, 262 ug/mL, and 4 Lig/mL curcumin, respec The same Volume, 1 mL, unless otherwise indicated, of tively (FIG. 4). Rubusoside was the best solubilizer for cur deionized, distilled water was added to each tube. Alterna 35 cumin. tively, a set percentage of water Solutions containing the solubilizer to be tested (e.g., 10% w/v) were prepared sepa Example 3 rately. In these cases, the solubilizer-water solutions were added directly to the tubes containing the low-solubility com Effect of Mogroside V. Geniposide, Paeoniflorin or pound. The tubes were then vortexed briefly and then soni 40 Rebaudioside A on the Water Solubility of cated for 60 minattemperature of 50° C. After sonication, the Camptothecin and Paclitaxel tubes were placed in an incubator set at 25°C. for at least 72 hr. The tubes were then centrifuged at 4000 rpm for 10 min. Ten percent w/v of mogroside V (1), geniposide (3), pae The Supernatant solution was passed through a 0.2 Lum filter oniflorin (4), rebaudioside A (8), or a water control containing and analyzed for the concentration of the low-solubility com 45 no solubilizing agents (9) water Solutions was each prepared. pound by HPLC or LC/MS. One set of two milligrams of camptothecin or paclitaxel were HPLC-UV and HPLC-MS Analysis: weighed into separate centrifuge tubes, and received one of The solutions containing various compounds in the the solubilizing agent 1, 3, 4, 8, or 9 (control). These water absence or presence of solubilizers were analyzed on HPLC solutions were sonicated at 50° C. for 60 min followed by UV or HPLC-MS which consisted of a solvent delivery pump 50 incubation at 25°C. for 72 hr. These compounds in the solu unit, an autosampler (Waters 717 plus), a UV-Vis diode array bilized water solutions were filtered by 0.45uM filters and detector (Waters 2996 Photodiode Array Detector, 190 to 800 analyzed on HPLC. Quantification was done by comparing to nm) coupled with an EMD 1000 Mass Detector (Waters), and a standard solution of a known amount in methanol. an evaporative light-scattering detector (Waters 2420 ELSD). Camptothecin. The system was computer controlled, and the results were 55 Chromatograms of five camptothecin water Solutions. analyzed using Empower software. Calibrations curves were CPT1, CPT3, CPT4, and CPT8 containing 10% w/v natural constructed using known concentrations of the compounds solubilizers (1-mogroside V, 3-geniposide, 4-paeoniflorin, and were used to quantify the concentrations of the com and 8-rebaudioside A) as complexing agents, respectively, are pounds dissolved in Solution. shown in FIG. 5. CPT9 was camptothecin dissolved in water 60 without any solubilizers (control). The HPLC system Example 2 included 600 pump, 717 autosampler, and 2996 PDA. Chro matograms were generated using a Prevail C18 column (2.1X Effect of Mogroside V. Astragaloside IV. Geniposide, 150 mm, 3 um) and a mobile phase of ACN (32): 0.02% or Paeoniflorin on the Water Solubility of Curcumin HCOOH H20 (68) at a flow rate of 0.4 mL/min, injection 65 volume of 1 uL, UV detection wavelength of 368 nm, and A 10% w/v of mogroside V (1), astragaloside IV (2), geni column temperature of 30° C. Camptothecin concentration poside (3), paeoniflorin (4), rubusoside (5), or Steviol mono was determined using a standard calibration curve with camp US 8,551,507 B2 17 18 tothecin standard solutions of 14, 70, and 140 ug/mL. The 10% geniposide; Tan-2 is the water solution containing 10% chromatograms above were generated at 368 nm UV showing paeoniflorin; Tan-3 is the water solution containing 10% elution of camptothecin at 7.3 min, the peak areas of which mogroside V: Tan-4 is the water solution containing 10% were used for quantification of camptothecin. The contents of astragaloside IV: Tan-5 is the water only solution (control): astragaloside and Steviol monoside in solutions were less than 5 and Ref is the reference standard compound of 30.4 ug/mL in 10%, as judged by their precipitations in the water solutions. methanol. In the presence of 10% w/v of mogroside V (1), geniposide In the presence of 10% w/v of mogroside V (1), astragalo (3), paeoniflorin (4), rebaudioside A or (8), the water solu side IV (2), geniposide (3), or paeoniflorin (4), the water tions contained 18 g/mL, 25ug/mL, 54 ug/mL, or 48 ug/mL Solutions contained 0.5 g/mL, O Lug/mL, 0.1 ug/mL, or 23 camptothecin, respectively (FIG. 5). The control water solu 10 ug/mL tanshinone IIA, respectively. The control water solu tion had no detectable level of camptothecin. tion had no detectable level of tanshinone IIA (FIG. 7). Paclitaxel: Coenzyme Q10: Chromatograms of five paclitaxel (Taxol) water Solutions. In the presence of 10% w/v of mogroside V (1), astragalo TXL1, TXL3, TXL4, and TXL8 containing 10% w/v natural side IV (2), geniposide (3), or paeoniflorin (4), none of the solubilizers (1-mogroside V, 3-geniposide, 4-paeoniflorin, 15 water solutions contained any detectable coenzyme Q10. The and 8-rebaudioside A) as complexing agents, respectively, are control water solution had no detectable level of coenzyme shown in FIG. 6. TXL9 was taxol dissolved in water without Q10 (Data not shown). any solubilizers (control). The HPLC-MS (Waters HPLC-MS Amphotericin B: system with 600 pump, 717 autosampler, 2996 PDA, and an Quantitative determination of amphotericin B is shown in EMD1000 MS detector) chromatograms were generated FIG. 8. All the chromatograms were detected at the wave using a Prevail C18 column (2.1 x 150 mm, 3 um) and a mobile length of 410 nm. A Luna C18 column (4.6 mmx250 mm, 5 phase of 0.25% HCOOH:ACN:MeOH (4:4:2 v/v/v); at a flow um) was used for the analyses. The mobile phase consisted of rate of 0.40 mL/min, injection volume of 1 uL., UV detection methanol (A) and water (B). Amphotericin B was eluted at wavelength of 230 nm, and column temperature of 30°C. MS 15.000 min. Amp-1 is the water solution containing 10% detection was performed with MS-ESI in positive mode and a 25 geniposide; Amp-2 is the water solution containing 10% pae SIR scan at m/z 854.4. Taxol concentration was determined oniflorin; Amp-3 is the water solution containing 10% using a standard calibration curve with taxol Standard Solu mogroside V: Amp-4 is the water solution containing 10% tions of 0.02, 0.06, and 0.2 mg/mL. The chromatograms astragaloside IV: Amp-5 is the water only solution (control); above were generated at SIR scan at m/z 854.4 showing and Ref is the reference standard compound of 55 ug/mL in elution of taxol at 8.1 min, the peak areas of which were used 30 methanol. for quantification of taxol. The contents of astragaloside and In the presence of 10% w/v of mogroside V (1), astragalo Steviol monoside in solutions were less than 10%, as judged side IV (2), geniposide (3), or paeoniflorin (4), the water by their precipitations in the water solutions Solutions contained 6 ug/mL, 0 ug/mL, 0.2 Lug/mL, or 19 In the presence of 10% w/v of mogroside V (1), geniposide ug/mL amphotericin B, respectively. The control water solu (3), paeoniflorin (4), or rebaudioside A (8), the water solu 35 tion had no detectable level of amphotericin B (FIG. 8). tions contained 7 ug/mL, 1 g/mL, 20 g/mL, or 26 ug/mL Artemisinin: paclitaxel, respectively (FIG. 6). The control water solution Quantitative determination of artemisinin is shown in FIG. had no detectable level of paclitaxel (FIG. 6). 9. All the chromatograms were detected at the wavelength of 205 nm. A Luna C18 column (4.6 mmx250 mm, 5um) was Example 4 40 used for the analyses. The mobile phase consisted of metha nol (A) and water (B). Artemisinin was eluted at 8.077 min. Effect of Mogroside V. Astragaloside IV. Geniposide, Art-1 is the water Solution containing 10% geniposide; Art-2 or Paeoniflorin on the Water Solubility of is the water solution containing 10% paeoniflorin; Art-3 is the Tanshinone IIa, Coenzyme Q10, Amphotericin b, water solution containing 10% mogroside V: Art-4 is the Artemisinin, Podophyllotoxin, Silybin, Propofol, and 45 water solution containing 10% astragaloside IV: Art-5 is the Celecoxib water only solution (control); and Ref is the reference stan dard compound of 730 ug/mL in methanol. A ten percent water solution w/v of mogroside V (1), In the presence of 10% w/v of mogroside V (1), astragalo astragaloside IV (2), geniposide (3), paeoniflorin (4), or a side IV (2), geniposide (3), or paeoniflorin (4), the water water control containing no solubilizing agents (5) was each 50 Solutions contained 148 g/mL, Oug/mL, 146 g/mL, or 218 prepared. One set of two milligrams oftanshinone IIA, coen ug/mL artemisinin, respectively. The control water Solution Zyme Q10, amphotericin B, artemisinin, podophyllotoxin, had 53 ug/mL of artemisinin (FIG. 9). Silybin, propofol, or celecoxib were each weighed into sepa Podophyllotoxin: rate centrifuge tubes, and received 1 mL of one of the solu Quantitative determination of podophyllotoxin is shown in bilizing agent 1, 2, 3, 4, or 5 (control). These water Solutions 55 FIG. 10. All the chromatograms were detected at the wave were sonicated at 50° C. for 60 min followed by incubation at length of 290 nm. A Luna C18 column (4.6 mmx250 mm, 5 25°C. for 72 hr. These compounds in the solubilized water um) was used for the analyses. The mobile phase consisted of solutions were filtered by 0.45 uM filters and analyzed on methanol (A) and water (B). Podophyllotoxin was eluted at HPLC. Quantification was done by comparing to a standard 13.220 min. Pod-1 is the water solution containing 10% geni Solution of a known amount in methanol. 60 poside: Pod-2 is the water solution containing 10% paeoni Tanshinone IIA: florin; Pod-3 is the water solution containing 10% mogroside Quantitative determination of tanshinone IIA is shown in V: Pod-4 is the water solution containing 10% astragaloside FIG. 7. All the chromatograms were detected using HPLC at IV: Pod-5 is the water only solution (control); and Ref is the the wavelength of 280 nm. A Luna C18 column (4.6 mmx250 reference standard compound of 370 ug/mL in methanol. mm, 5 um) was used for the analyses. The mobile phase 65 In the presence of 10% w/v of mogroside V (1), astragalo consisted of methanol (A) and water (B). Tanshinone IIA was side IV (2), geniposide (3), or paeoniflorin (4), the water eluted at 20.836 min. Tan-1 is the water solution containing Solutions contained 747 g/mL, Oug/mL, 644 g/mL, or 586 US 8,551,507 B2 19 20 ug/mL podophyllotoxin, respectively. The control water solu digoxin, oleandrin, nifedipine, or amiodarone were each tion had 53 ug/mL of podophyllotoxin (FIG. 10). weighed into separate centrifuge tubes, and received 1 mL of Silybin: one of the solubilizing agent 1,2,3,4,5,6,7,8, or 9 (control). Quantitative determination of silybin is shown FIG. 11. All These water solutions were sonicated at 50° C. for 60 min the chromatograms were detected at the wavelength of 288 followed by incubation at 25°C. for 72 hr. These compounds nm. A Luna C18 column (4.6 mmx250 mm, 5um) was used in the solubilized water solutions were filtered by 0.45 LM for the analyses. The mobile phase consisted of methanol (A) filters and analyzed on HPLC. Quantification was done by and water (B). Podophyllotoxin was eluted at 9.975 min. Sil-1 comparing to a standard solution of a known amount in is the water solution containing 10% geniposide; Sil-2 is the methanol. water solution containing 10% paeoniflorin; Sil-3 is the water 10 Clofazimine: solution containing 10% mogroside V: Sil-4 is the water Quantitative determination of clofazimine is shown in FIG. solution containing 10% astragaloside IV: Sil-5 is the water 14. All the chromatograms were detected at the wavelength of only solution (control); and Ref is the reference standard 289 mm. A Luna C18 column (4.6 mmx250 mm, 5um) was compound of 26 g/mL in methanol. used for the analyses. The mobile phase consisted of metha In the presence of 10% w/v of mogroside V (1), astragalo 15 nol (A) and water (B). Clofazimine was eluted at 15.000 min. side IV (2), geniposide (3), or paeoniflorin (4), the water C1 is the water solution containing 10% mogroside V: C2 is Solutions contained 18 g/mL, O Lug/mL, 16 ug/mL, or 37 the water solution containing 10% astragaloside IV: C3 is the ug/mL Silybin, respectively. The control water Solution had water Solution containing 10% geniposide; C4 is the water no detectable level of silybin (FIG. 11). solution containing 10% paeoniflorin: C5 is the water solu Propofol: tion containing 10% rubusoside; C6 is the water solution Quantitative determination of propofol is shown in FIG. containing 10% steviol monoside; C7 is the water solution 12. All the chromatograms were detected at the wavelength of containing 10% stevioside; C8 is the water solution contain 270 nm. A Luna C18 column (4.6 mmx250 mm, 5um) was ing 10% rebaudioside A: C9 is the water only solution (con used for the analyses. The mobile phase consisted of metha trol); and Ref is the reference standard compound of 160 nol (A) and water (B). Propofol was eluted at 13.084 min. 25 ug/mL in methanol. PRO-1 is the water solution containing 10% geniposide: In the presence of 10% w/v of mogroside V (1), astragalo PRO-2 is the water solution containing 10% paeoniflorin; side IV (2), geniposide (3), or paeoniflorin (4), rubusoside (5), PRO-3 is the water solution containing 10% mogroside V: Steviol monoside (6), Stevioside (7), or rebaudioside A (8), the PRO-4 is the water solution containing 10% astragaloside IV: water solutions contained 58 g/mL, 0.4 ug/mL, 48 Lug/mL, PRO-5 is the water only solution (control); and Ref is the 30 565 ug/mL, 132 ug/mL, Oug/mL, 64 Lug/mL, or 109 ug/mL reference standard compound of 702 ug/mL in methanol. clofazimine, respectively. The control water solution had 7.6 In the presence of 10% w/v of mogroside V (1), astragalo ug/mL of clofazimine (FIG. 14). side IV (2), geniposide (3), or paeoniflorin (4), the water Digoxin: Solutions contained 56 g/mL, O Lug/mL, 7 ug/mL, or 349 Quantitative determination of digoxin is shown in FIG. 15. ug/mL propofol, respectively. The control water solution had 35 All the chromatograms were detected at the wavelength of no detectable level of propofol (FIG. 12). 230 nm. A Luna C18 column (4.6 mmx250 mm, 5um) was Celecoxib: used for the analyses. The mobile phase consisted of metha Quantitative determination of celecoxib in FIG. 13. All the nol (A) and water (B). Digoxin was eluted at 16.927 min. D1 chromatograms were detected at the wavelength of 254 nm. A is the water solution containing 10% mogroside V: D2 is the Luna C18 column (4.6 mmx250 mm, 5um) was used for the 40 water solution containing 10% astragaloside IV: D3 is the analyses. The mobile phase consisted of methanol (A) and water Solution containing 10% geniposide; D4 is the water water (B). Podophyllotoxin was eluted at 12.051 min. Cel-1 is solution containing 10% paeoniflorin; D5 is the water solu the water solution containing 10% geniposide: Cel-2 is the tion containing 10% rubusoside; D6 is the water solution water solution containing 10% paeoniflorin: Cel-3 is the containing 10% steviol monoside; D7 is the water solution water solution containing 10% mogroside V: Cel-4 is the 45 containing 10% stevioside; D8 is the water solution contain water solution containing 10% astragaloside IV: Cel-5 is the ing 10% rebaudioside A: D9 is the water only solution (con water only solution (control); and Ref is the reference stan trol); and Ref is the reference standard compound of 388 dard compound of 84 g/mL in methanol. ug/mL in methanol In the presence of 10% w/v of mogroside V (1), astragalo In the presence of 10% w/v of mogroside V (1), astragalo side IV (2), geniposide (3), or paeoniflorin (4), the water 50 side IV (2), geniposide (3), or paeoniflorin (4), rubusoside (5), Solutions contained 47 g/mL, O Lug/mL, 3 ug/mL, or 253 Steviol monoside (6), Stevioside (7), or rebaudioside A (8), the ug/mL celecoxib, respectively. The control water Solution water Solutions contained 35ug/mL, 18 g/mL, 26 ug/mL, 38 had no detectable level of celecoxib (FIG. 13). ug/mL, 48 ug/mL, 10ug/mL, 30 ug/mL, or 32 ug/mL digoxin, respectively. The control water solution had 19 ug/mL of Example 5 55 digoxin (FIG. 15). Oleandrin: Effect of Geniposide, Paeoniflorin, Mogroside V. Quantitative determination of oleandrin is shown in FIG. Astragaloside IV. Rubusoside, Steviol Monoside, 16. All the chromatograms were detected at the wavelength of Stevioside, and Rebaudioside A on the Water 230 nm. A Luna C18 column (4.6 mmx250 mm, 5um) was Solubility of Clofazimine, Digoxin, Oleandrin, 60 used for the analyses. The mobile phase consisted of metha Nifedipine, and Amiodarone nol (A) and water (B). Oleandrin was eluted at 18.624 min. O1 is the water solution containing 10% mogroside V: O2 is Ten percent w/v of mogroside V (1), astragaloside IV (2), the water solution containing 10% astragaloside IV: O3 is the geniposide (3), paeoniflorin (4), rubusoside (5), Steviol water Solution containing 10% geniposide; O4 is the water monoside (6), Stevioside (7), rebaudioside A (8) or a water 65 solution containing 10% paeoniflorin; O5 is the water solu control containing no solubilizing agents (9) water Solutions tion containing 10% rubusoside; O6 is the water solution was each prepared. One set of two milligrams of clofazimine, containing 10% steviol monoside; O7 is the water solution US 8,551,507 B2 21 22 containing 10% stevioside; O8 is the water solution contain glycoside (rubusoside), and a tri-terpene glycoside (mogro ing 10% rebaudioside A, O9 is the water only solution (con side V) at 1:1:1 w/w/w ratio or a water control containing no trol); and Ref is the reference standard compound of 260 solubilizing agents water Solutions will be each prepared. ug/mL in methanol One set of two milligrams of curcumin, paclitaxel, camptoth In the presence of 10% w/v of mogroside V (1), astragalo 5 side IV(2), geniposide (3), or paeoniflorin (4), rubusoside (5), ecin, tanshinone IIA, digoxin, itraconazole, and celecoxib is Steviol monoside (6), Stevioside (7), or rebaudioside A (8), the each weighed into separate centrifuge tubes, and received 1 water Solutions contained 47 ug/mL, 5 g/mL, 28 Jug/mL, 42 mL of either the 10% combinational solubilizing agents or a ug/mL, 123 ug/mL, 7 ug/mL, 74 ug/mL, or 67 ug/mL olean water control. These water solutions are sonicated at 50° C. drin, respectively. The control water solution had 3 ug/mL of for 60 min followed by incubation at 25°C. for 72 hr. These oleandrin (FIG. 16). 10 compounds in the solubilized water solutions are filtered by Nifedipine: 0.45uM filters and analyzed on HPLC. Quantification is done Quantitative determination of nifedipine is shown in FIG. by comparing to a standard Solution of a known amount in 17. All the chromatograms were detected at the wavelength of methanol. It is expected that the combination of a monoter 235 nm. A Luna C18 column (4.6 mmx250 mm, 5um) was pene glycoside (paeoniflorin), a di-terpene glycoside (rubu used for the analyses. The mobile phase consisted of metha 15 nol (A) and water (B). Nifedipine was eluted at 20.871 min. Soside), and a tri-terpene glycoside (mogroside V) at 1:1:1 N1 is the water solution containing 10% mogroside V: N2 is w/w/w ratio will increase the solubility of curcumin, pacli the water solution containing 10% astragaloside IV: N3 is the taxel, camptothecin, tanshinone IIA, digoxin, itraconazole, water solution containing 10% geniposide: N4 is the water and celecoxib. solution containing 10% paeoniflorin; N5 is the water solu The complete disclosures of all references cited in this tion containing 10% rubusoside: N6 is the water solution specification are hereby incorporated by reference, including containing 10% steviol monoside; N7 is the water solution U.S. provisional patent application Ser. No. 61/219,973 and containing 10% stevioside; N8 is the water solution contain International Application No. PCT/US2009/040324, pub ing 10% rebaudioside A: N9 is the water only solution (con lished as WO2009/126950. In the event of an otherwise irrec trol); and Ref is the reference standard compound of 240 25 oncilable conflict, however, the present specification shall ug/mL in methanol. control. In the presence of 10% w/v of mogroside V (1), astragalo side IV(2), geniposide (3), or paeoniflorin (4), rubusoside (5), What is claimed: Steviol monoside (6), Stevioside (7), or rebaudioside A (8), the 1. A method for enhancing the solubility of an organic water Solutions contained 55ug/mL, 0 g/mL, 39 ug/mL, 49 30 compound which is insoluble or sparingly soluble in water, ug/mL, 286 g/mL, O Lug/mL, 468 ug/mL, or 203 Lug/mL said method comprising mixing said organic compound with nifedipine, respectively. The control water solution had no water and with a concentration of mogroside V sufficient to detectable level of nifedipine (FIG. 17). increase the solubility of the organic compound in water by a Amiodarone: factor of 2 or more above what the solubility would be in Quantitative determination of amiodarone is shown in FIG. 35 water without the mogroside V: wherein the organic com 18. All the chromatograms were detected at the wavelength of pound is selected from the group consisting of diterpenes, 242 nm. A Luna C18 column (4.6 mmx250 mm, 5um) was quinoline alkaloids, phenylalanine-derived alkaloids, used for the analyses. The mobile phase consisted of metha hydrolysable tannins, , curcuminoids, phenols, nol (A) and water (B). Amiodarone was eluted at 18.311 min. polymeric macrollides, cyclic peptides, sesquiterpene lac A1 is the water solution containing 10% mogroside V: A2 is 40 tones, lignans, flavonolignans, lipids, azoles, dihydropy the water solution containing 10% astragaloside IV: A3 is the ridines, amiodarones, and riminophenazines, wherein the water Solution containing 10% geniposide; A4 is the water concentration of mogrosideV is from about 1% to about 40% solution containing 10% paeoniflorin; A5 is the water solu weight/volume. tion containing 10% rubusoside; A6 is the water solution 2. The method of claim 1, wherein the organic compound is containing 10% steviol monoside; A7 is the water solution 45 selected from the group consisting of curcumin, camptoth containing 10% stevioside; A8 is the water solution contain ecin, paclitaxel, amphotericin B, artemisinin, podophyllo ing 10% rebaudioside A: A9 is the water only solution (con toxin, Silybin, propofol, celecoxib, clofazimine, oleandrin, trol); and Ref is the reference standard compound of 104 nifedipine, and amiodarone. ug/mL in methanol. 3. The method of claim 1, wherein the concentration of In the presence of 10% w/v of mogroside V (1), astragalo 50 mogroside V is about 10% w/v. side IV(2), geniposide (3), or paeoniflorin (4), rubusoside (5), 4. The method of claim 1, additionally comprising one or Steviol monoside (6), Stevioside (7), or rebaudioside A (8), the more solubilizing compounds to assist in Solubilizing the water Solutions contained 238 Lig/mL, 164 g/mL, 236 organic compound, wherein said solubilizing compound is ug/mL, 251 ug/mL, 249 ug/mL, Oug/mL, 201 ug/mL, or 271 selected from the group consisting of paeoniflorin, cyclodex ug/mL amiodarone, respectively. The control water Solution 55 trin, geniposide, rubusoside, Steviol monoside, Stevioside, had 107 ug/mL of amiodarone (FIG. 18). and rebaudioside A. 5. A method for enhancing the solubility of an organic Example 6 compound which is insoluble or sparingly soluble in water, said method comprising mixing said compound with water Effect of the Combination of a Monoterpene 60 and with geniposide in a concentration Sufficient to increase Glycoside, a Diterpene Glycoside, and a Triterpene the solubility of the organic compound in water by a factor of Glycoside on the Water Solubility of Curcumin, 2 or more above what the solubility would be in water without Paclitaxel, Camptothecin, Tanshinone IIA, Digoxin, the geniposide; wherein the organic compound is selected Itraconazole, and Celecoxib from the group consisting of quinoline alkaloids, phenylala 65 nine-derived alkaloids, hydrolysable tannins, flavonoids, cur Ten percent w/v of the combination of solubilizers consist cuminoids, phenols, cyclic peptides, sesquiterpene lactones, ing of a monoterpene glycoside (paeoniflorin), a di-terpene lignans, flavonolignans, lipids, azoles, dihydropyridines, US 8,551,507 B2 23 24 amiodarones, and riminophenazines, wherein the concentra amiodarones, and riminophenazines, wherein the concentra tion of geniposide is from about 1% to about 40% weight/ tion of rebaudioside A is from about 1% to about 40% weight/ Volume. Volume. 6. The method of claim 5, wherein the organic compound is 18. The method of claim 17, wherein the organic com selected from the group consisting of curcumin, camptoth pound is selected from the group consisting of clofazimine, ecin, artemisinin, podophyllotoxin, Silybin, propofol, cele oleandrin, nifedipine, and amiodarone. coxib, clofazimine, oleandrin, nifedipine, and amiodarone. 19. The method of claim 17, wherein the concentration of 7. The method of claim 5, wherein the concentration of rebaudioside A is about 10% w/v. geniposide is about 10% w/v. 20. The method of claim 17, additionally comprising one or 8. The method of claim 5, additionally comprising one or 10 more solubilizing compounds to assist in Solubilizing the more solubilizing compounds to assist in solubilizing the organic compound, wherein said solubilizing compound is organic compound, wherein said solubilizing compound is selected from the group consisting of mogroside V, cyclodex selected from the group consisting of mogroside V, cyclodex trin, paeoniflorin, geniposide, Steviol monoside, Stevioside, trin, paeoniflorin, rubusoside, Steviol monoside, Stevioside, 15 and rubusoside. and rebaudioside A. 21. A composition comprising an organic compound hav 9. A method for enhancing the solubility of an organic ing low solubility in water, and mogroside V: wherein the compound which is insoluble or sparingly soluble in water, organic compound is selected from the group consisting of said method comprising mixing said organic compound with diterpenes, quinoline alkaloids, phenylalanine-derived alka water and with a concentration of rubusoside sufficient to loids, hydrolysable tannins, flavonoids, curcuminoids, phe increase the solubility of the organic compound in water by a nols, macrollides, cyclic peptides, sesquiterpene lactones, lig factor of 2 or more above what the solubility would be in nans, flavonolignans, lipids, azoles, dihydropyridines, water without the rubusoside; wherein the organic compound amiodarones, and riminophenazines; and wherein the con is selected from the group consisting of cardiac glycosides, centration of said mogroside V in the composition is sufficient dihydropyridines, amiodarones, and riminophenazines, 25 to increase the solubility of said organic compound in water wherein the concentration of rubusoside is from about 1% to by a factor of 2 or more above what the solubility would be in about 40% weight/volume. an otherwise identical composition lacking said mogroside V. 10. The method of claim 9, wherein the organic compound and wherein the concentration of mogroside V is from about is selected from the group consisting of clofazimine, digoxin, 1% to about 40% weight/volume. oleandrin, nifedipine, and amiodarone. 30 22. The composition of claim 21, wherein the organic 11. The method of claim 9, wherein the concentration of compound is selected from the group consisting of curcumin, rubusoside is about 10% w/v. camptothecin, paclitaxel, amphotericin B, artemisinin, podo 12. The method of claim 9, additionally comprising one or phyllotoxin, Silybin, propofol, celecoxib, clofazimine, olean more solubilizing compounds to assist in solubilizing the drin, nifedipine, and amiodarone. organic compound, wherein said solubilizing compound is 35 23. The composition of claim 21, additionally comprising selected from the group consisting of mogroside V, cyclodex one or more solubilizing compound to assist in solubilizing trin, paeoniflorin, geniposide, Steviol monoside, Stevioside, the organic compound, wherein said solubilizing compound and rebaudioside A. is selected from the group consisting of paeoniflorin, cyclo 13. A method for enhancing the solubility of an organic dextrin, geniposide, rubusoside, Steviol monoside, Stevioside, compound which is insoluble or sparingly soluble in water, 40 and rebaudioside A. said method comprising mixing said compound with water 24. The composition of claim 21, additionally comprising and with a concentration of Stevioside Sufficient to increase one or more pharmaceutical agents selected from the group the solubility of the compound in water by a factor of 2 or consisting of complexing agents, cosolvents, Surfactants, more above what the solubility would be in water without the emulsifiers, liposomes and nanoparticles. Stevioside; wherein the organic compound is selected from 45 25. A composition comprising an organic compound hav the group consisting of phenazines and riminophenazines, ing low solubility in water, and geniposide; wherein the wherein the concentration of Stevioside is from about 1% to organic compound is selected from the group consisting of about 40% weight/volume. quinoline alkaloids, phenylalanine-derived alkaloids, 14. The method of claim 13, wherein the organic com hydrolysable tannins, flavonoids, curcuminoids, phenols, pound is selected from the group consisting of clofazimine, 50 cyclic peptides, sesquiterpene lactones, lignans, flavonolign oleandrin, and nifedipine. ans, lipids, azoles, dihydropyridines, amiodarones, and rimi 15. The method of claim 13, wherein the concentration of nophenazines; and wherein the concentration of said genipo Stevioside is about 10% w/v. side in the composition is sufficient to increase the solubility 16. The method of claim 13, additionally comprising one or of said organic compound in water by a factor of 2 or more more solubilizing compounds to assist in solubilizing the 55 above what the solubility would be in an otherwise identical organic compound, wherein said solubilizing compound is composition lacking said geniposide, and wherein the con selected from the group consisting of mogroside V, cyclodex centration of geniposide is from about 1% to about 40% trin, paeoniflorin, geniposide, Steviol monoside, rubusoside, weight/volume. and rebaudioside A. 26. The composition of claim 25, wherein the organic 17. A method for enhancing the solubility of an organic 60 compound is selected from the group consisting of curcumin, compound which is insoluble or sparingly soluble in water, camptothecin, artemisinin, podophyllotoxin, Silybin, propo said method comprising mixing said compound with water fol, celecoxib, clofazimine, oleandrin, nifedipine, and amio and with a concentration of rebaudioside A sufficient to darone. increase the solubility of the compound in water by a factor of 27. The composition of claim 25, additionally comprising 2 or more above what the solubility would be in water without 65 additionally comprising one or more solubilizing compound the rebaudioside A, wherein the organic compound is selected to assist in solubilizing the organic compound, wherein said from the group consisting of phenazines, dihydropyridines, solubilizing compound is selected from the group consisting US 8,551,507 B2 25 26 of mogroside V, cyclodextrin, paeoniflorin, rubusoside, Ste 34. The composition of claim 33, wherein the organic viol monoside, Stevioside, and rebaudioside A. compound is selected from the group consisting of clofaz 28. The composition of claim 25, additionally comprising imine, oleandrin, and nifedipine. one or more pharmaceutical agents selected from the group 35. The composition of claim 33, additionally comprising consisting of complexing agents, cosolvents, Surfactants, emulsifiers, liposomes and nanoparticles. one or more solubilizing compound to assist in solubilizing 29. A composition comprising an organic compound hav the organic compound, wherein said solubilizing compound ing low solubility in water, and rubusoside; wherein the is selected from the group consisting of mogroside V, cyclo organic compound is selected from the group consisting of dextrin, paeoniflorin, geniposide, Steviol monoside, rubusos cardiac glycosides, dihydropyridines, amiodarones, and rimi ide, and rebaudioside A. nophenazines; and wherein the concentration of said rubuso 10 36. The composition of claim 33, additionally comprising side in the composition is sufficient to increase the solubility one or more pharmaceutical agents selected from the group of said organic compound in water by a factor of 2 or more consisting of complexing agents, cosolvents, Surfactants, above what the solubility would be in an otherwise identical emulsifiers, liposomes and nanoparticles. composition lacking said rubusoside, and wherein the con 37. A composition comprising an organic compound hav centration of rubusoside is from about 1% to about 40% 15 weight/volume. ing low solubility in water, and rebaudioside A: wherein the 30. The composition of claim 29, wherein the organic organic compound is selected from the group consisting of compound is selected from the group consisting of clofaz phenazines, dihydropyridines, amiodarones, and rimi imine, digoxin, oleandrin, nifedipine, and amiodarone. nophenazines; and wherein the concentration of said rebau 31. The composition of claim 29, additionally comprising dioside A in the composition is sufficient to increase the one or more solubilizing compound to assist in Solubilizing solubility of said organic compound in water by a factor of 2 the organic compound, wherein said solubilizing compound or more above what the solubility would be in an otherwise is selected from the group consisting of mogroside V, cyclo identical composition lacking said rebaudioside A, and dextrin, paeoniflorin, geniposide, Steviol monoside, Stevio wherein the concentration of rebaudioside A is from about side, and rebaudioside A. 25 1% to about 40% weight/volume. 32. The composition of claim 29, additionally comprising 38. The composition of claim 37, wherein the organic one or more pharmaceutical agents selected from the group compound is selected from the group consisting of clofaz consisting of complexing agents, cosolvents, Surfactants, imine, oleandrin, nifedipine, and amiodarone. emulsifiers, liposomes and nanoparticles. 39. The composition of claim 37, additionally comprising 33. A composition comprising an organic compound hav 30 one or more solubilizing compound to assist in solubilizing ing low solubility in water, and stevioside; wherein the the organic compound, wherein said solubilizing compound organic compound is selected from the group consisting of is selected from the group consisting of mogroside V, cyclo phenazines and riminophenazines; and wherein the concen dextrin, paeoniflorin, geniposide, Steviol monoside, Stevio tration of said Stevioside in the composition is sufficient to side, and rubusoside. increase the Solubility of said organic compound in water by 35 40. The composition of claim 37, additionally comprising a factor of 2 or more above what the solubility would be in an one or more pharmaceutical agents selected from the group otherwise identical composition lacking said Stevioside, and consisting of complexing agents, cosolvents, Surfactants, wherein the concentration of Stevioside is from about 1% to emulsifiers, liposomes and nanoparticles. about 40% weight/volume. k k k k k