US006991800B2 United States Patent (12) (10) Patent N0.: US 6,991,800 B2 St0gniew (45) Date 0f Patent: Jan. 31, 2006 (54) ANTIFUNGAL PARENTERAL PRODUCTS 6,001,336 A 12/1999 Gordon 6,043,341 A 3/2000 Udodong et a1. (75) Inventor: Martin Stogniew, Blue Bell, PA (US) 6,153,224 A 11/2000 Stanifofth 6,258,821 B1 7/2001 Stogniew et a1. (73) Assignee: Vicur0n Pharmaceuticals Inc., gndlya etfil-t 1 , , ou oumie e a . Fremont’ CA(US) 6,284,282 B1 9/2001 Maa et al. ( * ) Notice: Subject' to any disclaimer, the term of this 22655550321151‘ patent is extended or ad]usted under 35 6,384,013 B1 5/2002 Burkhardt et aL U-S-C- 154(b) by 51 days~ 6,433,040 B1 8/2002 Dellamary et a1. 6,451,349 B1 9/2002 Robinson et al. (21) APP1- NO-I 10/172,678 6,475,523 B1 11/2002 Staniforth 6,506,726 B1 1/2003 Dobbins et al. (22) Filed: Jun. 13, 2002 6,565,885 B1 5/2003 Tarara et al. 6,590,073 B2 7/2003 Dalder et al. (65) Prior Publication Data 6,638,495 B2 10/2003 Weers et a1. 6,653,281 B1 11/2003 Borromeo et al. US 2004/0223997 A1 NOV. 11, 2004 6,660,843 B1 12/2003 Feige et aL 6,670,324 B2 12/2003 Jamison et a1. (51) Int- Cl- 6,689,390 B2 2/2004 Bernstein et a1. A61K 9/08 (2006.01) 6,709,650 B1 3/2004 Sutton et a1. A01N 25/04 (2006.01) 6,743,777 B1 6/2004 Burkhardt et a1. (52) US. Cl. ...................... .. 424/405; 424/400; 514/23; 2002/0151474 A1 10/2002 schwelr et al 514/359 2002/0160942 A1 10/2002 LareW et al. (58) Field of Classi?cation Search .............. .. 424/400, 22165:; al' 424/405; 514/3359 2003/0054981 A1 3/2003 Milton et a1. See application ?le for complete search history. 2003/0104048 A1 6/2003 Patel et a1_ 2003/0220236 A1 11/2003 Burkhardt et al. (56) References Cited FOREIGN PATENT DOCUMENTS U.S. PATENT DOCUMENTS CA 2043762 12/1991 3,293,482 A 12/1966 Wolkstein DE 28 03 851 A1 8/1979 3,978,210 A 8/1976 MiZuno et al. EP 0 031 221 7/1981 4,293,482 A 10/1981 Abbott et al. EP 0 032 009 A1 7/1981 4,293,483 A 10/1981 Debono EP 0 359 529 3/1990 4,293,489 A 10/1981 Debono EP 0 365 324 A1 4/1990 4,299,763 A 11/1981 Abbott et al. . 4,304,716 A 12/1981 Abbott et a1. (Commued) 44,,332408,,035824 AA 39// 11998822 AHborbioktots heti aelt. a1. 4,876,241 A 10/1989 Feldman et al. Avis, K. E. (1990). “Parenteral Preparations” Chapter 84 In 4,927,831 A 5/1990 Malamas Remington Pharmaceutical Sciences. 18th edition, Mack 5,141,674 A 8/1992 Leigh Publishing Company p. 1545-1569. 5,166,135 A 11/1992 SchmatZ _ 5,198,421 A 3/1993 Chen et al. (Con?rmed) 2 1532x2122 ee: :11" Printary Examiner—Leon B. Lankford, Jr. 5,541,160 A 7/1996 Balkovec et aL Assistant Examiner—Ruth A. Davis 5,573,936 A 11/1996 Kreuzman et a1_ (74) Attorney, Agent, or Firm—Morrison & Foerster LLP 5,618,787 A 4/1997 Jamison et a1. 5,629,289 A 5/1997 Rodriguez (57) ABSTRACT 5,629,290 A 5/1997 LaGrandeur et a1. _ _ _ _ _ 5,646,111 A 7/1997 Borromeo et aL Parenteral pharmaceutical formulations containing an ech1 5,652,213 A 7/1997 Jamison et a1_ nocandin antifungal compound and an aqueous solvent are 5,693,611 A 12/1997 Henle et a1, provided, Wherein the formulation includes ethanol, for 5,696,084 A 12/1997 Lartey et al. example about 20% W/v ethanol. The parenteral pharma 5,741,775 A 4/1998 BalkOVeC et 81- ceutical formulation may further include one or more addi 5786325 A 7/1998 Borromeo et al- tives, such as a stabilizing agent, buffer or tonicity agent. 5,932,543 A 8/1999 Burkhardt et a1‘ The parenteral pharmaceutical formulations are useful in 5’952’008 A 9/1999 Backstmm et a1‘ extending the shelf life and improving the solubility of the 5,965,525 A 10/1999 Burkhardt et a1. . . . 5,972,996 A 10/1999 Nielsen-Kahn et al. echmocandm annfungal Compound 5,985,309 A 11/1999 Edwards et al. 5,993,805 A 11/1999 Sutton et al. 48 Claims, N0 Drawings US 6,991,800 B2 Page 2 FOREIGN PATENT DOCUMENTS Ibrahim, F. S. et al., (1995) “the Effect of pH, sugars and calcium ion concentration on the thermal stability of Whey EP 0 365 324 B1 4/1990 proteins” Egyptian J. Dairy Sci. 23:177-178. EP 0 447 186 9/1991 EP 0 448 343 A2 9/1991 Longer, M. A. and Robinson, J. R. (1990). “Trasnsdermal EP 0 448 343 A3 9/1991 Systems” in Chapter 91 In Remington Pharmaceutical Sci EP 0 448 353 9/1991 ences. 18th edition, Mack Publishing Company. p. 1690 EP 0 448 354 A2 9/1991 1693. EP 0 448 354 A3 9/1991 Nail, S. and Gattlin, L. A. (1993). “Freeze Drying Principles EP 0 448 355 A2 9/1991 and Practice” Chapter 3 In Pharmaceutical Dosage forms, EP 0 448 355 A3 9/1991 EP 0 448 356 A2 9/1991 2nd edition . K. E. Avis ed. et al., Marcel Dekker, Inc. NY EP 0 448 356 A3 9/1991 pp. 163-233. EP 0 460 882 12/1991 Nema, S. et al. (1997). “EXcipients and Their Use in EP 0 462 531 12/1991 Injectable Products,” PDA Journal of Pharm. Science and EP 0 486 011 A2 5/1992 Tech. 51(4):166-171. EP 0 486 011 A3 5/1992 Sclarra, J. J. and Cutie, A. J. (1990). “Aerosols” Chapter 92 EP 0 503 960 A1 9/1992 EP 0 525 889 A1 2/1993 In Remington Pharmaceutical Sciences. 18th edition, Mack EP 0 561 639 9/1993 Publishing Company. p. 1694-1712. EP 0 589 074 3/1994 Turco, S. J. (1990). “Intravenous AdmiXtures,” Chapter 85 EP 0 744 405 11/1996 In Remington Pharmaceutical Sciences. 18th edition, Mack EP 0 757 058 2/1997 Publishing Company. p. 1570-1580. EP 0 931 834 7/1999 Groll, AH. et al. (2001). “Pharmacokinetic and GB 2241956 A 9/1991 Pharmacodynamic Modeling of Anidulafungin (LY303366): GB 2242194 A 9/1991 JP 03 240727 10/1991 Reappraisal of Its Ef?cacy in Neutropenic Animal Models of JP 05 271097 10/1993 Opportunistic Mycoses Using Optimal Plasma Sampling,” JP 5-271097 A 10/1993 Antimicrobial Agents and Chemotherapy 45910):2845 JP 06 172205 6/1994 2855. JP 6-172205 A 6/1994 International Search Report mailed on Dec. 9, 2003, for PCT W0 WO 94/25084 11/1994 WO WO-95/27074 A1 10/1995 Patent Application No. PCT/US03/18754 ?led on Jun. 12, W0 WO 96/31228 10/1996 2003, 5 pages. W0 WO 96/37509 11/1996 International Search Report for PCT Application No. PCT/ W0 WO 96/37510 11/1996 US00/05494 ?led Mar. 2, 2000, mailed Jun. 7, 2000, three W0 WO 96/37511 11/1996 pages. W0 WO 96/37512 11/1996 International Search Report for PCT Application No. PCT/ W0 WO 97/05163 2/1997 US00/00508 ?led Mar. 2, 2000, mailed Aug. 21, 2000, tWo W0 WO 97/27864 8/1997 W0 WO 97/30695 8/1997 pages. W0 WO 99/06062 2/1999 International Search Report for PCT Application No. PCT/ W0 WO 99/43337 9/1999 US00/05546 ?led Mar. 2, 2000, mailed Aug. 11, 2000, three W0 WO 00/11023 3/2000 pages. W0 W0 00/ 12540 3/2000 International Search Report for PCT Application No. PCT/ W0 WO 00/34315 6/2000 US00/05547 ?led Mar. 2, 2000, mailed Jul. 19, 2000, tWo W0 WO 00/35944 6/2000 pages. W0 WO 00/35945 6/2000 Keller-Juslen, C. et al. (1976). “Structure of the W0 WO 00/51564 9/2000 Cyclopeptide Antibiotic SL 7810 (=Echinocandin B),” W0 WO 00/51567 9/2000 Tetrahedron Letters 46:4147-4150. W0 WO 00/52036 9/2000 W0 WO 00/52037 9/2000 Turner, W.W. et al. (1996). “Recent Advances in the WO WO-03/105767 A2 12/2003 Medicinal Chemistry of Antifungal Agents,” Current WO WO-03/105767 A3 12/2003 Pharmaceutical Design 2:209-224. Debono, M. et al. (1995). “Semisynthetic Chemical OTHER PUBLICATIONS Modi?cation of the Antifungal Lipopeptide Echinocandin B Etter, MC. and Baures, PW. (1988) “Triphenylphosphine (ECB): Structure-Activity Studies of the Lipophilic and Oxide as a Crystallization Aid,”J. Am. Chem. Soc. 1101639 geometric Parameters of Polyarylated Acyl Analogs of 640. ECB,” JMed Chem. 38(17):3271-3281. US 6,991,800 B2 1 2 ANTIFUNGAL PARENTERAL PRODUCTS more additives, such as propylene glycol, or polyethylene glycol, a buffer, stabiliZing agent, tonicity agent, antioxidant TECHNICAL FIELD or bulking agent. Methods of reconstituting solid composi tions containing an echinocandin also are provided. This invention relates to drug formulations, for example, In one embodiment, pharmaceutically acceptable liquid aqueous injectable drug formulations and methods for their formulations are provided that include, e.g., about 0.2 to manufacture and use. 1.0%, or about 0.1 to 2.0% or about 0.2 to 2.0% W/v anidulafungin. The pharmaceutically acceptable liquid for BACKGROUND OF THE INVENTION mulations also may include, e.g., about 1.0 to 4.0 mg/mL, or about 1.5 to 3.0 mg/mL anidulafungin. 10 One aspect of the commercial viability of an injectable A pharmaceutically acceptable liquid parenteral formula drug product is long shelf life. A shelf life signi?cantly tion for example is provided comprising: anidulafungin and greater than one year is typically needed. This is because an aqueous solvent such as Water or saline, Wherein the drug products are often stored for long periods, for example, formulation includes from about 5% W/v ethanol to about six months to a year or more, until needed. The expiration 50% W/v ethanol; about 15—30% W/v ethanol; or about 20% date of a product begins When the drug is produced, but 15 W/v ethanol. The formulation may include about 10% to testing and packaging for shipping often take up some time, about 40% W/v ethanol and about 0.2 to about 2.0% W/v for example, months. A shelf life of one to three years or anidulafungin, or optionally about 15 to about 30% W/v more is very desirable for an injectable drug product. This is ethanol or optionally 20 percent W/v ethanol. The formula especially true for a drug Which may be stored for a long period of time, because it is not frequently used, but that is 20 tion also may include 10 to about 50 percent W/v propylene speci?cally required When indicated. glycol or polyethylene glycol, or mixtures thereof. Another aspect of injectable drug products is reconstitu The formulation further may include a stabiliZing agent, tion of the formulation by the medical practitioner. The drug such as mannitol, histidine, lysine, glycine, sucrose, fruc may be delivered in a solid form, often called “drug for tose, trehalose, lactose or a mixture thereof. The formulation injection,” Which may contain other ingredients, and is can optionally contain a bulking agent, such as mannitol, 25 reconstituted to a liquid form by the addition of solvent and glycine, lactose, sucrose, trehalose, dextran, hydroxyethyl other components. starch, ?coll or gelatin. The formulation can optionally A USP (United States Pharmacopeia) requirement for contain a solubiliZing agent or surfactant, such as cetrimide, parenteral drug products is that the product be visibly clear docusate sodium, glyceryl monooleate, sodium lauryl sul before use. A vial of crystal clear liquid is desired. To meet fate, or sorbitan esters. The solubiliZing agent or surfactant 30 this standard, the number of particulates in the reconstituted may optionally be a polyoxyethylenesorbitan fatty acid liquid product must be kept to a minimum. Particulates ester. Polyoxyethylenesorbitan fatty acid esters are also represent undissolved drug Which is ineffective, and may referred to as polysorbates, e.g., polysorbate 80 (polyoxy block capillaries causing serious adverse health effects. A ethylene sorbitan monooleate, TWeen 80), polysorbate 40 crystal clear drug product requires the solution to have a and polysorbate 20. Polysorbates, such as polysorbate 80, minimum of minute, undissolved, or non-visible drug par 35 are commercially available, for example, from Sigma, St. ticles. Louis, M0. Particulates in an injectable drug product may be caused The formulation can optionally comprise a buffer, such as in part by foaming during reconstitution. Foaming may be acetates, citrates, tartrates, lactates, succinates, or phos caused by the drug itself, or by surfactants other additives phates. The formulation can optionally contain a tonicity used to increase and hasten solubility of the drug. Foaming 40 agent, such as glycerin, lactose, mannitol, dextrose, sodium can prevent small particles from entering the solution to be dissolved, thereby increasing the number of particulates in chloride, sodium sulfate or sorbitol. The formulation can the reconstituted injectable drug product. optionally contain an antioxidant, such as acetone, sodium Another aspect of parenteral drug products is the time bisul?te, bisul?te sodium, butylated hydroxy anisole, buty required to reconstitute an injectable formulation. Reconsti 45 lated hydroxy toluene, cysteine, cysteinate HCl, dithionite tution requires the ability to rapidly redissolve a drug sodium, gentisic acid, gentisic acid ethanolamine, glutamate composition to provide a crystal clear solution. In addition, monosodium, formaldehyde sulfoxylate sodium, met the reconstitution should be rapid after a long storage period abisul?te potassium, metabisul?te sodium, monothioglyc of the delivered drug composition, a period Which can be, for erol, propyl gallate, sul?te sodium, thioglycolate sodium, or example, one, tWo, or three years or more. Some drug ascorbic acid. 50 compositions as delivered typically cannot achieve a shelf A pharmaceutically acceptable parenteral formulation is life greater than one year. This is because either the recon also provided for example comprising: anidulafungin and an stitution time is too long after storage, or the number of aqueous solvent, such as Water or saline, Wherein the for particulates in the reconstituted product is too high. mulation includes: Echinocandin antifungal compounds, and methods for about 5—30% W/v ethanol; 55 their manufacture and use are described, for example, in about 0.1—2.0% W/v anidulafungin; PCT WO 00/52037; PCT WO 00/51564; PCT WO about 0.1—1.0% W/v of a stabiliZing agent, such as fruc 00/34315; PCT WO 00/51567; and US. Pat. No. 5,965,525. tose; There is a need for echinocandin pharmaceutical drug about 0.1—10% W/v of a bulking agent, such as mannitol; formulations that are useful for parenteral pharmaceutical about 0.01—5% W/v of a buffer, such as tartaric acid; and administration With rapid reconstitution, little forming, and 60 about 0.1—5.0% W/v of a solubiliZing agent, such as a long shelf life. polysorbate 80. SUMMARY OF THE INVENTION The formulation may optionally include about 2—50% W/v polyethylene glycol and/or propylene glycol. In one aspect, a pharmaceutically acceptable parenteral 65 A pharmaceutically acceptable parenteral formulation is formulations containing an echinocandin and an aqueous also provided comprising: anidulafungin and an aqueous ethanolic solvent is provided, as Well as optionally one or solvent, such as Water or saline and ethanol, Wherein the US 6,991,800 B2 3 4 anidulafungin is stored in solid form for greater than 6, 9, 12, Also provided are pharmaceutically acceptable parenteral 15 or 18 months prior to forming the formulation, and formulations including anidulafungin and an aqueous sol Wherein the formulation is suitable for use as a parenteral vent, Wherein the anidulafungin is stored for greater than 6, formulation. 9, 12, 15, or 18 months before formation of the liquid A formulation is “suitable for use as a parenteral formu formulation, and Wherein the formulation is suitable for lation” if it is in a pharmaceutically acceptable form for administration as a parenteral formulation. parenteral administration. Thus, for example, for a liquid Further provided are methods of preparing a parenteral parenteral formulation, the particle content is sufficiently formulation, comprising combining a solid formulation loW, and the material is sufficiently sterile such that it is comprising anidulafungin With a solvent in an effective 10 useful for parenteral administration. To be suitable for amount to dissolve the anidulafungin rapidly, for example in parenteral administration, the formulation is visibly clear, 400 seconds or less, in 200 seconds or less, in 100 seconds and the number of particles in the reconstituted liquid or less, or in 60 seconds or less, for example by shaking or product is kept to a minimum. For example, less than 6,000 sWirling, to produce a pharmaceutically acceptable 10 pm particles should be present in a volume of 10 mL parenteral formulation. The anidulafungin may be a formu solvent that includes 35 mg of anidulafungin. For example, lation produced for pharmaceutical use and stored, for When the drug is freeZe dried and stored, for example, for 9 example for more than one year, or tWo years, prior to months, 12 months, 18 months or 24 months, and then combining With the solvent. The concentration of the anidu reconstituted by dissolving 35 mg of drug and optionally lafungin is, for example, about 1.5 to 3 mg/mL, or about 1.5 other additives in 10 ml of aqueous ethanolic solvent, there 20 to 5 mg/mL. are preferably less than 10,000, less than 6,000, less than 3,000, less than 1,000, or less than 400 10 pm particles. DETAILED DESCRIPTION OF THE There are, for example, less than 1000, less than 600, or less INVENTION than 200 25 pm particles in the 10 mL volume. Liquid parenteral formulations formed from drug stored 25 Provided are pharmaceutically acceptable formulations over long time periods can become no longer suitable for comprising an echinocandin, such as anidulafungin. parenteral administration, because of particles in the recon stituted formulations. Using the methods disclosed herein, Echinocandins liquid formulations are formed Which are suitable for Echinocandin-type compounds have been shoWn to parenteral administration, even When formed from drug 30 exhibit antifungal and antiparasitic activity, such as inhibi stored over long time periods. This can extend the shelf life tion of groWth of various infectious fungi including Candida of the drug. Moreover, using the ethanolic solutions for spp. (i.e., C Albicans, C Parapsilosis, C Krusei, C Glabrata, reconstitution results in more rapid dissolution of the drug. C Tropicalis, or C Lusitaniaw); Torulopus spp. (i.e., T Also provided is a method of preparing a parenteral Glabrata); Aspergillus spp. (i.e., A. Fumigalus), Histo 35 pharmaceutical formulation, comprising combining a solid plasma spp. (i.e., H. Capsulatum); Cryptococcus spp. (i.e., composition comprising anidulafungin and an ethanolic C. Neoformans); Blastomyces spp. (i.e., B. Dermatilidis); aqueous solvent, such as Water, to substantially dissolve the Fusarium spp.; Tr ichophyton spp., Pseudallescheria boydii, anidulafungin and produce an aqueous formulation includ Coccidioides immits, and Sporothrix schenckii, etc. PCT ing about 5 to about 50% W/v ethanol, optionally 10 to about 40 WO 00/51564. 30 percent W/v ethanol, Wherein the formulation is option ally shaken until the mixture is substantially clear. The Compounds of this type also have been shoWn to inhibit method optionally comprises forming the solid composition the groWth of certain organisms primarily responsible for containing anidulafungin by lyophiliZing an aqueous solu opportunistic infections in immunosuppressed individuals, tion of anidulafungin and optionally an additive such as such as groWth inhibition of Pneumocystis carinii. Other 45 polyethylene glycol or propylene glycol. The solid compo protoZoans that are inhibited by echinocandin-type com sition can optionally contain a stabiliZing agent, a buffer, a pounds include Plasmodium spp., Leishmania spp., Trypa tonicity agent or an antioxidant. nosoma spp., Cryptosporidium spp., Isospora spp., The method optionally comprises preparing a solid com Cyclospora spp., Trichomnas spp., and Microsporidiosis position comprising anidulafungin produced for pharmaceu 50 spp., etc. PCT WO 00/51564. tical use and stored for more than one year before combining Consequently, the formulations of the present invention With an ethanol Water solvent, and Wherein the combining are useful in the treatment of, e.g., systemic fungal infec step produces a formulation suitable for use as a pharma tions or fungal skin infections. Accordingly, the processes ceutically acceptable parenteral formulation. and formulations of the present invention may be used in the Also provided here is a kit for use in delivery of a 55 manufacture of a medicament for the therapeutic applica pharmaceutically acceptable parenteral pharmaceutical for tions described herein. For example, fungal activity (pref mulation, comprising: a vial containing a pharmaceutically erably, Candida albicans or Aspergillus fumigatis activity) acceptable solid formulation comprising anidulafungin; and or parasitic activity may be inhibited by contacting a phar a vial comprising a pharmaceutically acceptable aqueous maceutical formulation prepared by the present invention solution of about 10 to 30% W/v ethanol. The kit can 60 With a fungus or parasite, respectively. The term “contact optionally contain an additive, such as polyethylene glycol, ing” includes a union or junction, or apparent touching or propylene glycol, a stabiliZing agent, a buffer, a tonicity mutual tangency of a compound of the invention With a agent and/or an antioxidant. The kit can optionally contain parasite or fungus. The term does not imply any further a vial containing a solid formulation comprising 25 to 200 limitations to the process, such as by mechanism of inhibi mg of anidulafungin and a second vial containing 5 to 60 65 tion. The methods are de?ned to encompass the inhibition of milliliters of aqueous solution comprising about 10 to 30% parasitic and fungal activity by the action of the compounds W/v ethanol. and their inherent antiparasitic and antifungal properties. US 6,991,800 B2 5 6 Amethod for treating a fungal infection Which comprises Solid Compositions administering an effective amount of a pharmaceutical for Solid pharmaceutical compositions containing an echi mulation of the present invention to a host in need of such nocandin are provided that can be formulated for adminis treatment is also provided. The method includes treating a tration to a patient in need thereof. Such solid compositions Candida albicans or Aspergillus fumigatis infection. The may be reconstituted in an aqueous ethanolic solvent to provide a liquid product, Which may be administered to a term “effective amount” refers to an amount of active patient by parenteral means, including subcutaneous, intra compound Which is capable of inhibiting fungal activity. The venous, bolus injection, intramuscular, or intraarterial injec dose administered Will vary depending on such factors as the tion, or alternatively by oral, topical, transdermal, or nature and severity of the infection, the age and general 10 mucosal administration. health of the host and the tolerance of the host to the Solid compositions may have crystalline and amorphous antifungal agent. The particular dose regimen likeWise may components. Asolid composition of an echinocandin may be vary according to these factors. The drug may be given in a prepared by lyophiliZing (freeZe drying) a volume of a single daily dose or in multiple doses during the day. The solution Which contains a knoWn concentration of the echi regimen may last for example, for 2—3 days, for 14—30 days, 15 nocandin. or longer. An exemplary daily dose (administered in single A solid composition comprising anidulafungin can be or divided doses) contains a dosage level betWeen about 0.01 formed by lyophiliZing a solution of anidulafungin, such as mg/kg to 100 mg/kg of body Weight of an active compound. an aqueous solution of anidulafungin and optionally one or more additives, such as propylene glycol, or polyethylene Further exemplary daily doses are about 0.1 mg/kg to 60 20 glycol, a buffer, stabiliZing agent, tonicity agent, antioxidant mg/kg or about 0.1 mg/kg to 40 mg/kg, or about 0.7 to 3 or bulking agent. mg/kg per day. Further exemplary daily doses are about 5 to The polyethylene glycol may have, for example, a 500 mg/day or about 50 to 200 mg per day. molecular Weight of 400 to about 1500, optionally, 600 to Anidulafungin (1-[(4R,5R)-4,5-Dihydroxy-N(2)-[[4“ about 1000, often 1000. The addition of polyethylene glycol (pentyloxy)[1,1‘:4‘,1“-terphenyl]-4-yl]carbonyl]-L-orni 25 or propylene glycol optimiZes the reconstitution of the thine]echinocandin B) is an echinocandin that can be semi lyophiliZation formulation in an aqueous solvent containing synthetically derived from a natural product. The synthesis ethanol by providing enhanced solubility. The amount of of anidulafungin is described in US. Pat. No. 5,965,525. polyethylene glycol or propylene glycol is, for example, an Like other echinocandins, anidulafungin has a loW Water amount effective to produce the desired concentrations after solubility of less than 0.1 mg/ml. Because of the loW 30 addition of an aqueous solvent to form a parenteral formu solubility, formulations have been described that add a lation as discussed herein. In the solid composition, for surfactant to an aqueous solution, hoWever this can hinder example, a 50 mg dose of anidulafungin may contain 1.5 freeZe drying (WO 00/51564). grams of PEG. It has been found in the past to be difficult to prepare a Solid formulations may optionally contain a stabiliZing formulation of anidulafungin in Water Which meets the strict 35 agent. The solid formulations may contain a stabiliZing USP requirements for purity and clarity of injectable for reagent at a concentration of 5 to 80% W/W, generally 7.6 to mulations, even When such concentrations are Well beloW 11.5% W/W, or 9.5% W/W. The term “stabilizing agent” refers the solubility limit in the presence of surfactant species. to a pharmaceutically acceptable excipient that may enhance The structure of anidulafungin is provided beloW the chemical or physical stability of the active ingredient in ZE null/OH Anidulafungin US 6,991,800 B2 7 8 the formulation. Suitable stabilizing agents include carbo solvents, solvent mixtures, or solvent systems are revieWed hydrates, such as sucrose, trehalose, fructose, and lactose, in, S. Nema et al, PDA Journal ofPharm. Science and Tech. and amino acids. Other examples include polyoxyethylene 51(4): 166—171 (1997). The solution to be lyophiliZed may sorbitan fatty esters (polysorbates), e.g., polysorbate 80 be steriliZed prior to lyophiliZation. Alternatively, the solid (polyoxyethylene sorbitan monooleate, TWeen 80), polysor product from lyophiliZation may be steriliZed. Methods for bate 40 and polysorbate 20. steriliZation are revieWed in Remington’s Pharmaceutical Solid formulations may optionally contain a solubiliZing Sciences, 18th ed. (1990). agent or surfactant. The solid formulations may contain a Asuitable method for freeZe-drying is described in Nail et solubiliZing agent at a concentration of 10 to 50% W/W, 20 al. Freeze Drying Principles and Practice, in Pharmaceutical to 30% W/W, or e.g., 24% W/W. Suitable solubiliZing agents 10 Dosage Forms, 2” Ed., Marcel Dekker, Inc. NY, pp. include cetrimide, docusate sodium, glyceryl monooleate, 163—233 (1993). The formulation is preferably freeZe-dried sodium lauryl sulfate, and sorbitan esters. Exemplary solu in a vial Which can then be stored until needed. Anon-toxic, biliZing agents include polysorbates (e.g. polysorbate 20, aqueous solvent is added to the vial to dissolve the freeZe polysorbate 40, polysorbate 80). dried material thus forming a solution that can be used in a As used herein, “W/W” refers to percent Weight in Weight, 15 parenteral therapeutic application. Those skilled in the art and expresses the number of g of a constituent in 100 g of Will appreciate that the aqueous solvent includes other solution or mixture. common solutions used in such applications (e.g., saline As used herein, “W/v” refers to percent Weight in volume solutions, dextrose solutions, etc.). In application, the for and expresses the number of g of a constituent in 100 mL of mulations are typically diluted or reconstituted (if freeZe solution. 20 dried) and may be further diluted if necessary, prior to Solid formulations may also optionally contain a buffer. administration. The buffer is optionally present at a concentration in the The active ingredient is typically formulated into phar range from about 0.3 to 5%, or about 0.9% to 1.3%, or about maceutical dosage forms to provide an easily controllable 1.1% W/W. The term “buffer” refers to a pharmaceutically dosage of the drug and to give the patient an elegant and acceptable excipient that helps to maintain the pH of the 25 easily handleable product. Solid formulations may comprise solution Within a particular range speci?c to the buffering for example, about 0.1% to 60% W/W of active ingredient for system. Suitable buffers include acetates, citrates, phos example, anidulafungin, or about 1% to 30% W/W, or about phates, tartrates, lactates, succinates, amino acids and the 8.0 to 12% W/W, or optionally about 9—10% W/W. like. As used herein, the term “unit dose” or “unit dosage” When freeZe dried, the formulations may optionally con 30 refers to physically discrete units that contain a predeter tain a bulking agent. The term “bulking agent” refers to a mined quantity of active ingredient calculated to produce a pharmaceutically acceptable excipient that adds bulk to a desired therapeutic effect. When a unit dose is administered formulation Which results in a Well-formed cake upon freeZe parenterally, it is typically provided in the form of measured drying. Suitable bulking agents include mannitol, glycine, units in ampoules (or vials). The dosage to be administered lactose, sucrose, trehalose, dextran, hydroxyethyl starch, 35 may vary depending upon the physical characteristics of the ?coll and gelatin. The bulking agent is for example present patient, the severity of the patient’s symptoms, and the in the formulation at the concentration in the range from means used to administer the drug. The speci?c dose for a about 30 to 68% W/W of the solid formulation. For example, given patient is usually set by the judgment of the attending the formulation comprises about 43 to 52% and optionally physician. Dosages can be for example, about 5—500 mg or about 48% W/W bulking agent 40 about 35—200 mg daily administered from an intravenous The solid formulations may be prepared using conven injection (IV). tional dissolution and mixing procedures. For example, the Suitable carriers, diluents and excipients are knoWn in the anidulafungin is dissolved in a non-toxic aqueous solvent art and include materials such as carbohydrates, Waxes, optionally in the presence of a pharmaceutically acceptable Water soluble and/or sWellable polymers, hydrophilic or polyethylene glycol or propylene glycol and optionally one 45 hydrophobic materials, gelatin, oils, solvents, Water, and the or more bulking agents, buffers, and/or stabiliZing agents. like. The particular carrier, diluent or excipient used Will The resulting solution is then steriliZed, e.g., sterile ?ltered depend upon the means and purpose for Which the active and preferably lyophiliZed to provide the desired formula ingredient is being applied. The formulations may also tion. include Wetting agents, lubricating agents, emulsi?ers, sus The solution to be lyophiliZed may further include one or 50 pending agents, preservatives, sWeeteners, perfuming more antioxidants, such as acetone sodium bisul?te, bisul?te agents, ?avoring agents and combinations thereof. sodium, butylated hydroxy anisole, butylated hydroxy tolu Shelf life of the solid compositions is the length of time ene, cystein, cysteinate HCl, dithionite sodium, gentisic that the solid composition may be stored in a form intended acid, gentisic acid ethanolamine, glutamate monosodium, for a parenteral preparation and still be reconstituted in a formaldehyde sulfoxylate sodium, metabisul?te potassium, 55 reasonable time. The shelf life can begin When the active metabisul?te sodium, monothioglycerol, propyl gallate, drug ingredient of the solid composition is made, and can sul?te sodium, thioglycolate sodium, and ascorbic acid. end When the solid composition cannot be reconstituted in a The solid composition also may include a tonicity agent reasonable time by a particular method or When the drug such as glycerin, lactose, mannitol, dextrose, sodium chlo degrades or otherWise cannot be used. Solid compositions ride, sodium sulfate and sorbitol. 60 With long shelf life, for example, greater than 12 months; or The solution to be lyophiliZed may include excipients, greater than about 15, 20, 25, 30, 35, 36 months or more are stabiliZing agents, buffers, tonicity agents, and antioxidants, preferred. As disclosed herein, the solvent added to the solid as described above, and may further contain agents Which composition to make the parenteral formulation can increase modify the physical appearance and shape of the lyophiliZed the shelf life, to, for example, greater than 12 months; or solid such as mannitol, fructose, and tartaric acid. Solid 65 greater than 15, 20, 25, 30, 35, 36 months or more. compositions obtained from lyophiliZation of the solution An example of solid formulations for a 35 and 50 mg dose may include amounts of all species described herein. Further of anidulafungin is shoWn in Table A. This Table shoWs an US 6,991,800 B2 9 10 exemplary amount as Well as an optional range shown in aqueous ethanol solution. These formulations may be further parentheses of components of a solid formulation. These diluted With 5% Dextrose in Water (D5W) prior to intrave formulations are made in one embodiment by dissolving the nous (IV) injection. ingredients in sterile Water (e. g., 3—15 ml), optionally adjust The pharmaceutical parenteral formulations may be pro ing the pH With and freeZe drying. vided in dosage vessels Which contain dosage units, for example, about 5 mg to 500 mg of anidulafungin, about 20 TABLE A to 200 mg, or about 35 mg, 50 mg, or 100 mg. The dosage vessels are often loaded With a slight excess, for example Solid Anidulafungin Formulations 2.5% excess of drug, because it is generally not possible to 10 remove all the drug from the vial after reconstitution of the Quantity in Quantity drug. For example, a 35 mg vessel may be loaded With about mg in mg per per vial 50 vial 35 mg 36 mg of drug. A dosage unit may be provided in a sealed mg dose dose % W/W container, often made of Type I glass, Which maintains a Ingredient (mg range) (range) Function (range) sterile environment for the solid product from lyophiliZa Anidulafungin 50 35 Active 9.5 15 tion. For example, a vial hermetically sealed With a sterile (40-60) (28-42) Ingredient (7.6—11.5) rubber or plastic closure or stopper. The closure or stopper Fructose 50 35 Stabiliser 9.5 alloWs charging the vial With solvent or diluent, such as (45-55) (31.5-38.5) (86-105) sterile Water for Injection, USP, ethanol, Normal Saline, Mannitol 250 175 Bulking Agent 48 (225-275) (157-192) (43-52) USP, or 5% Dextrose in Water, USP. Polysorbate 80 125 87.5 mg Solubilizing 24 20 A pharmaceutically acceptable parenteral liquid product (113-137) (79-96) Agent/ (21-26) formulation may be prepared by reconstitution of a solid surfactant composition such as a freeZe dried solid composition, for Tartaric Acid 5.6 3.94 Buffer 1.1 example, as described above. These product liquids may be (5.0-6.1) (3.5-4.3) (0.9-1.3) Sodium As needed To adjust pH negligible steriliZed during, or after reconstitution. The reconstitution Hydroxide 25 product may be used to parenterally administer an echi Hydrochloric As needed To adjust pH negligible nocandin active compound, such as anidulafungin. Recon Acid stitution may be accomplished by charging a vessel con taining the solid composition With a physiologically The solid or liquid formulations in a freeZe dried dosage acceptable sterile solvent or diluent and mixing the contents vial may include an excess of about 1—5%, e.g., 2.5% of any 30 of the vessel for an acceptable reconstitution time by hand one or more of the above components to alloW for With shaking or sWirling. Other methods of shaking or mixing the draWal of the required amount of the anidulafungin from the vessel may be used, for example, any mechanical shaking or reciprocating mixing device. The intensity of shaking or vial after addition of solvent and extraction for parenteral mixing is sufficient to reconstitute the solid composition into administration. 35 a liquid product in a reasonable time, for example, Within a In application, the formulations are reconstituted (if lyo feW minutes, for example, Without causing severe foaming. philiZed) and are further diluted prior to administration. An The reconstitution time of the solid composition, When example of reconstitution instructions for a freeZe-dried reconstituted With one of the aqueous formulations compris product is to add aqueous ethanolic solvent to the vial and ing ethanol described herein, may be much less than the agitate to dissolve. Typical reconstitution times are less than 40 reconstitution time of the solid composition When reconsti ten minutes, or less than one minute The resulting solution tuted With Water. The reconstitution may be carried out With may be then further diluted in an infusion solution such as an aqueous solution and ethanol as Well as one or more dextrose 5% in Water (D5W), prior to intravenous admin additives, to provide a pharmaceutically acceptable istration. parenteral formulations. Apharmaceutical composition may be administered using 45 Reconstitution time is, for example, less than 10 minutes, a variety of methods. Suitable methods include injection. less than 5, minutes, less than 2 minutes, less than one The particular treatment method used Will depend upon the minute, or less than 45 seconds, for example after storage of type of infection being addressed. a freeZe dried drug formulation for more than one year, more Liquid Pharmaceutical Formulations than tWo years or more than three years. Preferably, the Provided are parenteral pharmaceutical formulations 50 liquid drug formulation after reconstitution includes no more than a pharmaceutically acceptable amount of particulate comprising anidulafungin and an aqueous solvent, Wherein matter. For example, the content of particulate matter after the formulation includes from about 5% ethanol W/W to dissolving 35 mg of drug in 10 ml of aqueous ethanolic about 50 percent ethanol W/W. The aqueous solvent, is for solvent, after freeZe drying and storage, for example, after example, Water, or e.g. saline. 55 one, tWo or three years, is less than 6000, preferably less As used herein, “W/W” refers to percent Weight in Weight, than 400 10 pm particles, and/or less than 600, preferably and expresses the number of g of a constituent in 100 g of less than 200 25 pm particles. solution or mixture. The addition of surfactant or solubiliZing agent, for Also provided are pharmaceutically acceptable parenteral example polyoxyethylene sorbitan monooleate (TWeen 80 or formulations comprising anidulafungin and an aqueous sol 60 polysorbate 80), improves solubility but can cause foaming, vent, Wherein the formulation includes about 5% to about Which reduces exposure of the drug to the solution and can 50% W/W ethanol, e.g., about 10 to 40%; about 15 to 30%; cause less drug to be dissolved. Insolubility problems or about 20% W/W ethanol. increase With increasing storage time of a freeZe dried Also provided are pharmaceutically acceptable parenteral material. Particles can be dispersed on top of the solution in formulations comprising anidulafungin and an aqueous sol 65 the bubbles of the foam Which makes it difficult to see in the vent, Wherein the formulation includes about 5 to 50% (W/W) vial if the drug is dissolved. For example, When TWeen 80 propylene glycol and/or polyethylene glycol (PEG) in an is added to increase the solubility of anidulafungin, and the US 6,991,800 B2 11 12 drug is freeze dried to enhance stability, the ability to redissolve the freeze dried formulation in Water becomes TABLE C-continued increasingly reduced over time. The formulations of the present invention overcome this disadvantage. The ethanol Optional Ranges of Liquid Formulations of Anidulafungin in the reconstitution solution can act by reducing the surface % tension of the reconstituted drug formulation resulting in Wgt./Wgt. reduced foaming. This procedure also reduces reconstitution Ingredient range time, due to the solubility properties of ethanol. Sodium Hydroxide — Another advantage is that reconstitution is greatly Hydrochloric Acid — improved in solid formulations stored longer than one year. 10 Ethanol 5-30 The surfactants become less effective over time and PEG and/or propylene glycol 5-30 increased foaming is observed in reconstituted drug formu Sterile Water for Injection to volume lations stored 18 months or longer using Water for recon stitution Without ethanol. This may restore surfactant func In a pharmaceutically acceptable aqueous parenteral for tion of stored solid drug formulations. 15 mulation of an echinocandin, such as anidulafungin, the In one embodiment, the pharmaceutically acceptable Weight percent of polyethylene glycol or propylene glycol, parenteral formulation of echinocandin, such as anidulafun if present, may be, e.g., 5 to 50%; or about 20% W/v in an gin, may have about 5 to 50% W/v ethanol, e. g., ethanol USP, aqueous solvent such as Water or an Water-ethanol mixture. in Water, or 20% W/v ethanol in Water or other aqueous The molecular Weight of the polyethylene glycol may be solution. In these pharmaceutically acceptable parenteral 20 less than about 1500, often less than about 1000, sometimes aqueous formulations, the % W/v ethanol, such as ethanol less than about 600, and sometimes less than about 400. USP may be, for example, about 5 to about 50 percent W/v; Polyethylene glycol having molecular Weight in the range eg about 10 to about 40%; about 15 to about 30%; about 18 400-600 may be optimal. In a pharmaceutically acceptable to 22 percent; or about 20 percent W/v ethanol. parenteral formulation of echinocandin, such as anidulafun Tables B and C shoW examples of liquid formulations of 25 gin, the Weight percent of polyethylene glycol may be for anidulafungin, Where the formulations include, Water, etha example about 5 to 50% W/v or about 10 to 20% W/v in an nol, anidulafungin and optionally one or more of the other aqueous solvent comprising ethanol. components listed in the Tables; as Well as exemplary ranges A pharmaceutically acceptable parenteral formulation of of the components. These liquid formulations may be further echinocandin such as anidulafungin can include about 10% diluted in D5W or other aqueous diluent for intravenous 30 W/v ethanol and 10% W/v propylene glycol or polyethylene injection. glycol in an aqueous solution such as Water. In the pharma ceutically acceptable parenteral formulations, in one TABLE B embodiment the sum of the percent W/v is less than or equal to 50. Liquid Formulations of Anidulafungin 35 The formulation can for example include an echinocandin such as anidulafungin at a concentration of about 0.1 to 2.0% Quantity % per vial W/v W/v or about 0.25 to 0.45% W/v or optionally 0.34% W/v. Ingredient 35 mg dose in 10 ml (range) (range) In reconstitution of the solid composition With an aqueous ethanol mixture after a long storage period of eg greater Anidulafungin 35 mg 0.34 (0.1-2.0) 40 than one year, tWo years, three years or more, the mixing or Fructose 35 mg 0.34 shaking time for reconstitution may be less than about 400 (0.1-1.0) seconds, often less than about 300 seconds, sometimes less Mannitol 175 mg 1.75 than about 200 seconds, sometimes less than about 100 (0.1-10) seconds, and sometimes less than about 75 seconds, Wherein Polysorbate 80 87.5 mg 0.85 (Tween 80) (0.1-5.0) 45 results may be improved in comparison to results using Tartaric Acid 3.94 mg 0.04 aqueous solvents Without ethanol. (0.01-50) Formulations may optionally contain a stabiliZing agent. Sodium Hydroxide As needed negligible Hydrochloric Acid As needed negligible A stabiliZing agent is present optionally at a concentration Ethanol 2 g 20 for example, in the range of about 0.3% to about 40% W/v, (5-50) 50 or about 1% to about 10% W/v. The term “stabiliZing agent” PEG and/or 1 g 10 refers to a pharmaceutically acceptable excipient that propylene glycol (0-50) enhances the chemical or physical stability of the active Sterile Water for q.s to 10 ml to volume Injection ingredient in the formulation. Suitable stabiliZing agents include carbohydrates (e.g., as sucrose, trehalose, fructose, 55 lactose and mannitol), and amino acids. Formulations may also optionally contain a solubiliZing TABLE C agent. The solubiliZing agent is present, for example, at a Optional Ranges of Liquid Formulations of Anidulafungin concentration of about 0.2% to about 2.0% W/v, or about 0.75% to about 1.0% W/v. Suitable solubiliZing agents include polysorbates (eg polysorbate 20, polysorbate 40, and 60 Wgt./Wgt. polysorbate 80). Ingredient range Formulations may also optionally contain a buffer. The Anidulafungin 0.27-0.42 buffer is present for example at a concentration in the range Fructose 0.3—0.39 from about 0.03% to about 5.0% W/v, or about 0.1% to about Mannitol 1. 6-1.9 Polysorbate 80 0.75-0.96 65 2.0% W/v. The term “buffer” refers to a pharmaceutically Tartaric Acid 0033-0043 acceptable excipient that maintains the pH of the solution Within a particular range speci?c to the buffering system. US 6,991,800 B2 13 14 Formulations may have, for example, a pH of about 3.0—7.0, The folloWing examples are provided to illustrate but not optionally about 4.0—6.0, or about 4.4—4.6. Suitable buffers limit the invention. include acetates, citrates, phosphates, tartrates, lactates, suc All documents, including publications, treatises, articles, cinates, amino acids and the like. Sodium hydroxide and/or patents and patent applications referenced herein are incor hydrochloric acid can be used to adjust the pH. porated herein by reference in their entirety. Formulations may optionally contain a bulking agent. The bulking agent is for example present in a formulation at a EXAMPLES concentration in the range from about 1% to about 60% W/v, Example 1 or about 3% to about 50% W/v. The term “bulking agent” refers to a pharmaceutically acceptable excipient that adds 10 Method of Manufacture of Solid Anidulafungin bulk to a formulation Which results in a Well-formed cake upon freeZe drying. Suitable bulking agents include manni Formulation tol, glycine, lactose, sucrose, trehalose, dextran, hydroxy ethyl starch, ?coll and gelatin. Preferred bulking agents Manufacture include mannitol, sucrose, trehalose, lactose and combina 15 In a suitable manufacturing vessel, 25 grams of polysor tions thereof. bate 80, are added to a sufficient amount of deoxygenated Formulations may further include one or more antioxi Sterile Water for Injection and mixed sloWly until dissolved. dants. The antioxidant is for example present in a formula The solution is cooled and 1.1 gram of tartaric acid is added. tion at a concentration range of 0.01—10% W/v or about The solution is adjusted to pH 4.5 using a sodium hydroxide 1—5% W/v. Examples of antioxidants include acetone 20 solution and/or a hydrochloric acid solution. sodium bisul?te, bisul?te sodium, butylated hydroxy ani 10 grams of anidulafungin are added to a suitable vessel sole, butylated hydroxy toluene, cystein, cysteinate HCl, With Sterile Water for Injection and sWirled to generate a dithionite sodium, gentisic acid, gentisic acid ethanolamine, slurry. The resulting slurry is added to the bulk polysorbate glutamate monosodium, formaldehyde sulfoxylate sodium, 80 buffer solution and the liquid is mixed until all the metabisul?te potassium, metabisul?te sodium, mono 25 slurried drug is dissolved. The fructose (10 grams) and thioglycerol, propyl gallate, sul?te sodium, thioglycolate mannitol (50 grams) are added and mixed until dissolved. sodium, and ascorbic acid. Additional Sterile Water for Injection is added to bring the In application, the formulation is typically reconstituted solution to ?nal volume. The pH of the ?nal solution is and further diluted if necessary, prior to administration. An determined and adjusted, if necessary, to pH 4.5. The example of reconstitution instructions for the lyophiliZed 30 solution is steriliZed by membrane ?ltration using tWo product are to add solvent to the vial and agitated to dissolve. redundant in-line 0.22M Millipak 20 ?lters into a Class 100 Typical reconstitution times are less than 400 seconds. aseptic area. These membrane ?lters are non-asbestos, non Suitable solvents include ethanol in Water. In one example, ?ber releasing, and meet cGMP requirements for use in the manufacture and processing of components of drug products the solvent is about 20 Weight percent ethanol in an aqueous solution such as Water. The resulting solution is optionally 35 for parenteral injection in humans. SteriliZing ?lters are further diluted in an infusion solution such as dextrose 5% integrity tested after use to assure that integrity Was main in Water (D5W), prior to administration. tained during ?ltration. In-process testing occurs on the bulk material after the ?ltration for Appearance, Anidulafungin A solid composition may be reconstituted to provide a concentration, and pH. liquid product for parenteral administration to a patient. 40 Solid compositions to be reconstituted may be provided in LyophiliZation dosage vessels Which contain dosage units, for example, Aliquots (3.5 ml) of the sterile ?ltered bulk solution are from about 5 mg to about 500 mg of echinocandin, such as aseptically ?lled into the sterile (depyrogenated) glass vials anidulafungin, for example about 15 mg, about 25 mg, about and sterile stoppers are partially inserted. The ?lled vials are 35 mg, about 50 mg, about 100 mg, or about 200 mg. The 45 transferred to the freeZe dryer and lyophiliZed. dosage vessels are often loaded With a slight excess, for The freeZe drying process is monitored by thermocouple example 2.5% excess of drug, because it is generally not probes of representative vials. Vials are loaded into the possible to extract all the drug upon use of the reconstituted freeZe dryer and the temperature is gradually reduced until drug. For example, a 35 mg vessel may be loaded With about all thermocouples reach —40° C. After the desired time has 36 mg of drug. A dosage unit may be provided in a sealed 50 elapsed, the air in the chamber is evacuated and the tem container, often made of Type I glass, Which maintains a perature gradually increased until the shelf temperature is sterile environment for the solid product from lyophiliZa approximately +35° C. The samples are held at about +35° tion, for example, a 50 ml vial hermetically sealed With a C. for 6—8 hours. The chamber is then restored to atmo sterile rubber or plastic closure or stopper. spheric pressure With ?ltered Nitrogen UHP and the stoppers The solid product, for example, 35 mg of anidulafungin, 55 are seated. Vials are removed and capped With aluminum and optionally other components of the formulation, in a 10 seals. mL vial, or 50 mg anidulafungin in a 15 mL vial, can be During aseptic operations such as set-up, ?ltration, ?lling reconstituted in a pharmaceutically acceptable diluent, for and stoppering, the air is monitored for microbial content example 5—50% W/v ethanol or 20% W/v ethanol in Sterile With settling plates and volumetric air sampler. In addition, Water for Injection (SWFI) to a concentration of drug that is 60 operators and surfaces are monitored by contact plates. The for example about 0.5—5 mg/mL, about 3—4 mg/ml or about air is monitored for particles using a particle counter. 3.3 mg/ml. The reconstituted drug is then further diluted Records of the results of these surveys are evaluated against about 5—10 fold or about 7 fold With a pharmaceutically pre-established action limits for the area involved and, if acceptable diluent such as 5% Dextrose in Water (D5W), to necessary, an investigation is conducted. Appropriate action a concentration of, e.g., 0.1 to 3 mg/mL or about 0.5 mg/mL 65 is taken When indicated by the results of the investigation. for administration. This solution can be administered intra All components and equipment are steriliZed by appro venously. priate processes. SteriliZation processes uses the “overkill” US 6,991,800 B2 15 16 approach for both steam and dry heat sterilization cycles. By shaking the diluent, reconstitution times are shortened, The cycle for steam autoclaving is for 30 minutes at 121° C. but foaming can often occur With the use of Water alone. and the cycle for dry heat sterilization is for at least three Using aqueous ethanol as a diluent, it is possible to use hours at >250° C. The resulting solid is stored at room shaking to dissolve the drug quickly, and foaming is temperature. 5 reduced. Reconstitution As illustrated in Table 1, reconstitution of solid compo Reconstitution occurs in a tWo step process. In the ?rst sitions containing anidulafungin Was signi?cantly reduced step, the solid formulation containing 35 mg anidulafungin, from greater than 5400 seconds to 74 seconds using 20% and optionally other components of the formulation, is W/W ethanol in Water in comparison With pure Water. 10 reconstituted in a 10 ml solution of 20% W/v ethanol in Sterile Water for Injection. The mixture is sWirled by hand TABLE 1 for 100 seconds and observed to con?rm the complete dissolution of the solid product in a clear bubble free Reconstitution Times of Solid Compositions Containing solution. This mixture is then diluted 7 fold in a solution of Anidulafungin Shaken in Indicated Diluent 15 5% Dextrose in Sterile Water (D5W). The resulting solution Storage Storage Timei is noW available for IV injection. Condition Lot No. Time (Seconds) diluent" Example 2 Initial CT 1 2758 Initial 24 Water 25° C./60% RH CT12758 3 month 164 Water 25° C./60% RH CT12758 18 month 338 Water Reconstitution Time of Solid Anidulafungin 20 25° C./60% RH CT12758 25 month >5400 Water Compositions 25° C./60% RH CT12758 36 month >5400** Water Initial PPD04365 Initial 27 Water 25° C./60% RH PPD04365 3 month 101 Water A: Reconstitution Time of Anidulafungin Stored for up to 36 25° C./60% RH PPD04365 18 month 271 Water 25° C./60% RH PPD04365 25 month >5400 Water Months 25 Initial CT12759 Initial 32 Water Solid compositions containing 1) 35 mg anidulafungin 25° C./60% RH CT12759 3 month 64 Water (lot CT12759, PPD04365), 175 mg mannitol USP, 87.5 mg 25° C./60% RH CT12759 18 month 247 Water polysorbate 80 NF, 35 mg fructose USP, and 3.95 mg tartaric 25° C./60% RH CT12759 25 month >5400** Water 25° C./60% RH CT12759 36 month >5400** Water acid NF as a buffer; or 2) 25 mg (lot CT12758) anidulafun 25° C./60% RH CT12759 32 months 74 Aqueous Ethanol gin, 125 mg mannitol USP, 62.5 mg polysorbate 80 NF, 25 30 mg fructose USP, and 2.5 mg tartaric acid NF as a buffer, ivalues represent the average of 5 repeats Were tested to determine reconstitution time after storage. *Water is sterile Water ?ltered for injection (SWFI); aqueous ethanol is 20% W/W ethanol in SWFI. The samples of lyophiliZed formulations, obtained from Eli **Method modi?cation: mechanical platform shaker used, room tempera Lilly (Indianapolis, Ind.), Were stored in solid form for up to ture 36 months or longer. Storage conditions Were 250 C., or 50 35 RH = relative humidity C. 60% relative humidity The reconstitution time assay consisted of initiating reconstitution of the solid Tables 2 and 3 shoW a comparison of reconstitution times composition by injecting 10 ml of solvent into the vial for different lots of solid anidulafungin compositions in containing 35 mg of anidulafungin and 7 ml of solvent into Water and aqueous ethanol, Where, advantageously, drug the vial containing 25 mg of anidulafungin. Once the solvent 40 could be dissolved in 120 seconds or less With shaking using Was added, it Was immediately mixed in the vial by gently aqueous ethanol. Generally using Water, shaking produces shaking or sWirling by hand. Every 10 seconds shaking Was unWanted foaming. SWirling can be used to reduce foaming, stopped and the vial Was visually inspected until complete but the length of reconstitution time can increase consider ness of solution Was observed. The results in Table 1 shoW ably. Reconstitution time using Water and shaking varied each reconstitution time reported represented as an average 45 betWeen 150 and 360 seconds. Reconstitution times using of 5 replicates. It can be seen from the Tables that the Water and sWirling, varied betWeen 240 and 1200 seconds. addition of ethanol to the reconstitution mixture signi? The reconstitution time using Water and ethanol and shaking cantly reduces the reconstitution time of anidulafungin. is betWeen 60 and 120 seconds and is signi?cantly reduced Acomparison Was made betWeen shaken and sWirled (by relative to reconstitution in Water. Thus, using a Water/ hand) reconstitution methods. SWirling generally results in ethanol mixture results less foaming and a reduced recon longer reconstitution times, but is used to reduce foaming. stitution time. TABLE 2 Reconstitution Times for Lot 12758 Stored for 3 Years at 5° C. Time Lot No. Vial Diluent" Agitation (seconds) Final Reconstitution Evaluation SWirled CT 12758 1 Water SWirled 1200 Hazy, With a signi?cant number of small particles CT 12758 2 Water SWirled 600 Completely in solution CT 12758 3 Water SWirled 1200 Clear solution With a small number of large particles