WO2006094029A2

WO2006094029A2 - Pharmaceutical formulations containing 17-allylamino-17-demethoxygeldanamycin

Info

Publication number: WO2006094029A2
Application number: PCT/US2006/007210
Authority: WO
Inventors: Indu Isaacs; John G. Augustine; Usha Srinivasula
Original assignee: Kosan Biosciences Incoporated
Priority date: 2005-02-28
Filing date: 2006-02-28
Publication date: 2006-09-08
Also published as: CA2596867A1; EP1853238A2; WO2006094029A3; JP2008531708A; EP1853238A4

Abstract

A pharmaceutical formulation comprising 17-AAG in an amount from about 1 mg/mL to about 5 mg/mL dissolved in a vehicle comprising (i) a first component that is an aprotic, polar solvent in an amount between about 0.1 and about 10 volume %; and, (ii) a second component that is an aqueous mixture comprising between about 5.0 and about 55.0 volume % long chain triglycerides, in an amount between about 90.0 and 99.9 volume %.

Description

PHARMACEUTICAL FORMULATIONS CONTAINING 17-ALLYLAMINO- 17-DEMETHOXYGELDANAMYCIN

BACKGROUND OF THE INVENTION

1. FIELD OF THE INVENTION

[0001] This invention relates to pharmaceutical formulations containing 17-allylamino-17- demethoxygeldanamycin ("17-AAG") and methods for their preparation and use.

2. DESCRIPTION OF RELATED ART

[0002] Geldanamycin belongs to the ansamycin family of natural products, whose members are characterized by a benzenoid nucleus (typically a benzoquinone or hydroquinone nucleus) connected at two meta positions to form a macrolactam. Besides geldanamycin, the ansamycins include the macbecins, the herbimycins, the TAN-420s, and reblastatin. [0003] Geldanamycin and its derivatives are the most extensively studied of the ansamycins. Although geldanamycin was originally identified as a result of screening for antibiotic activity, current interest in it is based primarily on its cytotoxicity towards tumor cells and, therefore, its potential as an anticancer agent. It is an inhibitor of heat shock protein-90 ("Hsp90"), which is involved in the folding, activation and assembly of a wide range of proteins ("client proteins"), including key proteins involved in signal transduction, cell cycle control and transcriptional regulation. The binding of geldanamycin to Hsp90 disrupts Hsp90-client protein interactions, preventing the client proteins from folding correctly and rendering them susceptible to proteasome-mediated destruction. Among the Hsp90 client proteins are many mutated or overexpressed proteins implicated in cancer: p53, Bcr-Abl lcinase, Raf-1 kinase, Akt kinase, Npm-Alk kinase pl85^ErB2 transmembrane kinase, Cdk4, Cdk6, Weel (a cell cycle-dependent lcinase), Her2/Neu (ErbB2), and hypoxia inducible factor- lα (HIF-lα). However, the hepatotoxicity and poor bioavailability of geldanamycin have led to its discontinuation as a clinical candidate.

[0004] Nevertheless, interest persists in the development of geldanamycin derivatives or analogs having geldanamycin-like bioactivity, but with a more pharmaceutically acceptable spectrum of properties. Position 17 of geldanamycin has been an attractive focal point, chemically speaking, for the synthesis of geldanamycin derivatives because the methoxy group there is readily displaced by a nucleophile, providing a convenient entry into 17- substituted-17-demethoxygeldanamycins. Further, structure-activity relationship (SAR) studies have shown that chemically and sterically diverse 17-substituents can be introduced without destroying antitumor activity. See, e.g., Sasaki et al., US 4,261,989 (1981); Schnur et al, US 5,932,566 (1999); Schnur et al, J. Med. Chem., 38, 3806-3812 (1995); Schnur et al, 3 Med. Chem. 38, 3813-3820 (1995); and Santi et at., US 2003/0114450 Al (2003); the disclosures of which are incorporated by reference. The SAR inferences are supported by the X-ray crystal co-structure of the complex between Hsρ90 and a geldanamycin derivative (17- DMAG, v. infra), showing that the 17-substituent projects out from the binding pocket and into the solvent (Jez et ah, Chemistry & Biology, 10, 361-368 (2003)). Thus, position 17 is an attractive one for the introduction of property-modulating substituents, such as a solubilizing group. The best-known 17-substituted geldanamycin derivative is 17-AAG, first disclosed in Sasaki et at., cited supra, and currently undergoing clinical trials. Another noteworthy 17-substituted geldanamycin compound is 17-(2-dimethylaminoethyl)amino-17- demethoxygeldanamycin (("17-DMAG") (Snader et al., WO 02/079167 Al (2002), incorporated by reference), also undergoing clinical trials. [0005] A limitation in the preparation of pharmaceutical formulations containing geldanamycin compounds such as geldanamycin itself and 17-AAG, especially for parenteral administration, is their very poor water solubility, only about 0.1 mg/mL at neutral pH for 17- AAG. (17-DMAG, having an alkyl amino group, is more soluble.) Addressing this issue, Tabibi et al., WO 00/37050 (2000) disclosed a formulation for a water insoluble drug such as 17-AAG comprising (a) the drug, (b) a water-miscible organic solvent for the drug, (c) a surfactant, and (d) water. The water miscible solvent can be dimethylsulfoxide (DMSO), dimethylformamide, ethanol, glycerin, propylene glycol, or polyethylene glycol. The surfactant preferably is a phospholipid (especially egg phospholipid).

[0006] Another disclosure of interest is UIm et al., WO 03/086381 (2003), which discloses a method for preparing pharmaceutical formulations for ansamycins by (a) providing the ansamycin dissolved in ethanol; (b) mixing the product of step (a) with a medium chain triglyceride to form a first mixture; (c) substantially removing the ethanol from the first mixture; (d) combining the product of step (c) with an emulsifying agent and a stabilizer to form a second mixture; and (e) emulsifying the second mixture. The emulsified second mixture optionally can be lyophilized and then re-hydrated. In a specific combination, the medium chain triglyceride comprises caprylic and/or caproic acid, the emulsifying agent comprises phosphotidylcholine, and stabilizer comprises sucrose. The aforementioned documents are incorporated herein by reference.

BRIEF SUMMARY OF THE INVENTION [0007] The present invention provides a pharmaceutical formulation for 17-AAG, a method for administering 17-AAG to a patient in need thereof, and a method for preparing a pharmaceutical formulation comprising 17-AAG.

[0008] In one aspect, the present invention provides a formulation including 17-AAG that is suitable for intravenous administration. The formulation comprises 17-AAG in a concentration of between about 1.0 mg/mL and about 5.0 mg/mL dissolved in a vehicle comprising (i) a first component that is an aprotic, polar solvent in an amount between about 0.1 and about 10 volume %; and, (ii) a second component that is an aqueous mixture comprising between about 5.0 and about 55 volume % long chain triglycerides, in an amount between about 90.0 and 99.9 volume %.

[0009] In another aspect, the present invention provides a method for administering 17-AAG to a patient in need thereof, comprising the steps of:

(a) providing a pharmaceutical formulation comprising 17-AAG in a concentration of between about 1.0 mg/mL and about 5.0 mg/mL dissolved in a vehicle comprising (i) a first component that is an aprotic, polar solvent in an amount between about 0.1 and about 10 volume %; and, (ii) a second component that is an aqueous mixture comprising between about 5.0 and about 55 volume % long chain triglycerides, in an amount between about 90.0 and 99.9 volume %.

(b) including the pharmaceutical formulation into an apparatus adapted to deliver it intravenously to a patient through an intravenous route; and,

(c) administering the formulation intravenously to a patient.

[0010] In yet another aspect, the present invention provides a method for preparing a pharmaceutical formulation comprising 17-AAG, comprising the steps of:

(a) providing an amount of 17-AAG;

(b) combining the 17-AAG with a first component, which is an aprotic, polar solvent, to provide a solution of the 17-AAG in the aprotic, polar solvent; and,

(c) combining the solution with a second component, wherein the second component comprises an aqueous mixture comprising between about 5.0 and about 55.0 volume % long chain triglycerides in an amount between about 90.0 and 99.9 volume % of the final formulation; thereby forming the pharmaceutical formulation.

DETAILED DESCRIPTION OF THE INVENTION

Definitions [0011] Aprotic, polar solvents are solvents that do not contain an O-H group but still possess a fairly large dipole. Examples of aprotic, polar solvents include, without limitation, dimethylsulfoxide (i.e., DMSO), N,N-dimethylacetamide (i.e., DMA), N,N- dimethylformamide (i.e., DMF), and N-methylpyrrolidone (i.e., NMP).

[0012] Intralipid, either 10% or 20%, includes soybean oil, phospholipids (lecithin), glycerin and water for injection. One liter of Intralipid 10% contains the following: 100 g of purified soybean oil; 12 g of purified egg phospholipids, 22 g anhydrous glycerin, and water to a volume of 1 liter. The pH is adjusted with sodium hydroxide to approximately 8. One liter of Intralipid 20% contains the following: 200 g of purified soybean oil; 12 g of purified egg phospholipids, 22 g anhydrous glycerol, and water to a volume of 1 liter. The pH is adjusted with sodium hydroxide to approximately 8. The Intralipids are typically stored at a controlled temperature below 25 ⁰C.

[0013] Liposyn II, either 10% or 20%, includes safflower oil, soybean oil, phospholipids

(lecithin), glycerin and water for injection. Liposyn II 10% contains 5 weight % safflower oil, 5 weight % soybean oil, up to 1.2 weight % egg phosphatides added as an emulsifier, and

2.5 weight % glycerin in water for injection. The pH may be adjusted with sodium hydroxide to between 6 and 9. Liposyn II 20% contains 10 weight % safflower oil, 10 weight % soybean oil, 1.2 weight % egg phosphatides, and 2.5 weight % glycerin in water for injection..

The pH may be adjusted with sodium hydroxide to between 6 and 9.

[0014] Lecithin is a mixture of the diglycerides of stearic, palmitic, and oleic acids, linked to the choline ester of phosphoric acid. Commercial lecithin is typically soybean lecithin, which contains 11.7% palmitic acid, 4.0% stearic acid, 8.6% palmitoleic acid, 9.8% oleic acid,

55.0% linoleic acid, 4.0% linolenic acid, and 5.5% C₂₀ to C₂₂ acids. Egg lecithins are one suitable type of lecithin.

[0015] Linoleic acid is a fatty acid of the formula C₁₈H₃₂O₂.

[0016] Linolenic acid is a fatty acid of the formula C₁₈Hs₀O₂.

[0017] Long chain triglycerides are triglyceride compositions that include fatty acids ranging from 12 to 22 carbons in length as the predominant constituent, preferably 16 to 20 carbons in length.

[0018] Medium chain triglycerides are triglyceride compositions that include fatty acids ranging from 7 to 11 carbons in length as the predominant constituent, preferably 8 to 10 carbons in length.

[0019] Oleic acid is a fatty acid of the formula C₁₈H₃₄O₂.

[0020] Palmitic acid is a fatty acid of the formula C₁₆H₃₂O₂. [0021 ] Safflower oil is a mixture of triglycerides of palmitic acid (6.4%), stearic acid (3.1%), arachidic (0.2%), oleic acid (13.4%), linoleic acid (76.6-79.0%), and linolenic acid (0.04-

0.13%).

[0022] Sesame oil comprises primarily triglycerides of oleic and linoleic acids. However, it appeal's to adversely affect the stability of 17-AAG and is not as desirable for this reason.

[0023] Stearic acid is a fatty acid of the formula C₁₈H₃₆O₂.

[0024] Soybean oil is a mixture of triglycerides of oleic acid (26%), linoleic acid (49%) linolenic acid (11%), and saturated acids (14%).

[0025] Soybean oil/safflower oil in a 50/50 mixture includes triglycerides composed of approximately 65.8% linoleic acid, 17.7% oleic acid, 8.8% palmitic acid, 3.4% stearic acid, and 4.2% linolenic acid.

Description

[0026] The pharmaceutical formulation of the present invention includes 17-AAG and is suitable for intravenous administration. The formulation comprises 17-AAG in a concentration of between about 1.0 mg/mL and about 5.0 mg/mL dissolved in a vehicle comprising (i) a first component that is an aprotic, polar solvent in an amount between about

0.1 and about 10 volume %; and, (ii) a second component that is an aqueous mixture comprising between about 5.0 and about 55.0 volume % long chain triglycerides, in an amount between about 90.0 and 99.9 volume %.

[0027] The concentration of 17-AAG in the formulation is typically between about 1.25 mg/mL and 4.0 mg/mL. Where the aprotic, polar solvent in the formulation is DMSO, the concentration of 17-AAG oftentimes ranges from about 2.0 mg/mL to about 3.0 mg/mL; where the aprotic, polar solvent is DMA, the concentration oftentimes ranges from about 1.50 mg/mL to about 3.0 mg/mL.

[0028] Aprotic, polar solvents are typically present in the formulation in an amount between about 0.5 and about 5.0 volume %. The concentration of such solvents is maintained within

FDA acceptable limits due to toxicity at higher concentrations. For example, the tolerated dose for DMA is approximately 14.8 g/m². Oftentimes, the concentration of aprotic, polar solvent ranges from about 1.0 to about 4.0 volume %.

[0029] The second component of the vehicle is typically an aqueous mixture comprising between about 5.0 and about 55.0 volume % long chain triglycerides. Examples of suitable second components include, without limitation, the following: Intralipid 10%; Intralipid 20%; Liposyn II 10%; and Liposyn II 20%. For these mixtures, the triglyceride content oftentimes ranges from about 7.5 to about 30 volume %.

[0030] Phospholipids, preferably egg phospholipids, are an optional third component of the vehicle. Where the second component is Intralipid or Liposyn II, the phospholipid concentration oftentimes ranges from about 0.8 to about 3.0 volume percent, with 1.0 to 2.0 volume percent being preferred.

[0031] Examples of suitable 17-AAG of the present invention include, without limitation, the following:

[0032] Formulation 1 — comprising a) concentration of 17-AAG of between about 1 mg/mL and about 3 mg/mL; b) first component is DMSO present in an amount between about 1.0 to about 4.0 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0033] Formulation 2 — comprising a) concentration of 17-AAG of between about 2.0 mg/mL and about 3.0 mg/mL; b) first component is DMSO present in an amount between about 1.0 to about 4.0 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0034] Formulation 3 — comprising a) concentration of 17-AAG of between about 2.25 mg/mL to about 2.75 mg/mL; b) first component is DMSO present in an amount between about 1.0 to about 4.0 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0035] Formulation 4 — comprising a) concentration of 17-AAG of between about 2.25 mg/mL to about 2.75 mg/mL; b) first component is DMSO present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0036] Formulation 5 — comprising a) concentration of 17-AAG of between about 2.25 mg/mL to about 2.75 mg/mL; b) first component is DMSO present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides.

[0037] Formulation 6 — comprising a) concentration of 17-AAG of between about 2.25 mg/mL to about 2.75 mg/mL; b) first component is DMSO present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid, oleic acid and palmitic acid.

[0038] Formulation 7 — comprising a) concentration of 17-AAG of between about 2.25 mg/mL to about 2.75 mg/mL; b) first component is DMSO present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid (50% to 75% of fatty acids) , oleic acid (15% to 25% fatty acids) and palmitic acid (7% to 10% fatty acids).

[0039] Formulation 8 — comprising a) concentration of 17-AAG of between about 2.25 mg/mL to about 2.75 mg/mL; b) first component is DMSO present in an amount between about 2.0 to about 3.5 volume %; c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid (50% to 75% of fatty acids) , oleic acid (15% to 25% fatty acids) and palmitic acid (7% to 10% fatty acids); and, d) third component — phospholipids — present in an amount between about 1.0 percent and 2.0 volume percent.

[0040] Formulation 9 — comprising a) concentration of 17-AAG of between about 2.25 mg/mL to about 2.75 mg/mL; b) first component is DMSO present in an amount between about 2.0 to about 3.5 volume %; c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid (50% to 75% of fatty acids) , oleic acid (15% to 25% fatty acids) and palmitic acid (7% to 10% fatty acids); d) third component — egg phospholipids — present in an amount between about 1.0 w and 2.0 volume percent; and, e) glycerin in an amount between about 2.0 and 3.0 volume %. [0041 ] Formulation 10 — comprising a) concentration of 17-AAG of between about 1.0 mg/mL and about 3.0 mg/mL; b) first component is DMA present in an amount between about 1.0 to about 4.0 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0042] Formulation 11 — comprising a) concentration of 17-AAG of between about 1.5 mg/mL and about 3 mg/mL; b) first component is DMA present in an amount between about 1.0 to about 4.0 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0043] Formulation 12 — comprising a) concentration of 17-AAG of between about 1.65 mg/mL to about 2.50 mg/mL; b) first component is DMA present in an amount between about 1.0 to about 4.0 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0044] Formulation 13 — comprising a) concentration of 17-AAG of between about 1.65 mg/mL to about 2.50 mg/mL; b) first component is DMA present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 7.5 and 30.0 volume % long chain triglycerides.

[0045] Formulation 14 — comprising a) concentration of 17-AAG of between about 1.65 mg/mL to about 2.50 mg/mL; b) first component is DMA present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides.

[0046] Formulation 15 — comprising a) concentration of 17-AAG of between about 1.65 mg/mL to about 2.50 mg/mL; b) first component is DMA present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid, oleic acid and palmitic acid.

[0047] Formulation 16 — comprising a) concentration of 17-AAG of between about 1.65 mg/mL to about 2.50 mg/mL; b) first component is DMA present in an amount between about 2.0 to about 3.5 volume %; and, c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid (50% to 75% of fatty acids) , oleic acid (15% to 25% fatty acids) and palmitic acid (7% to 10% fatty acids).

[0048] Formulation 17 — comprising a) concentration of 17-AAG of between about 1.65 mg/mL to about 2.50 mg/mL; b) first component is DMA present in an amount between about 2.0 to about 3.5 volume %; c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid (50% to 75% of fatty acids) , oleic acid (15% to 25% fatty acids) and palmitic acid (7% to 10% fatty acids); and, d) third component — phospholipids — present in an amount between about 1.0 and 2.0 volume percent.

[0049] Formulation 18 — comprising a) concentration of 17-AAG of between about 1.65 mg/mL to about 2.50 mg/mL; b) first component is DMA present in an amount between about 2.0 to about 3.5 volume %; c) second component is an aqueous mixture comprising between about 15.0 and 30.0 volume % long chain triglycerides, wherein the fatty acid component of the triglycerides comprises linoleic acid (50% to 75% of fatty acids) , oleic acid (15% to 25% fatty acids) and palmitic acid (7% to 10% fatty acids); d) third component — egg phospholipids — present in an amount between about 1.0 and 2.0 volume percent; and, e) glycerin in an amount between about 2.0 and 3.0 volume %.

[0050] The formulations of the present invention exhibit stability for a period of hours at a variety of temperatures. For instance, the 17-AAG of Formulations 1, 2, 3, 4, 5, 6, 7, 8, and 9 — at room temperature, 4 ⁰C, and -20 ⁰C — degrades less than 5% over a period of 7 hours, preferably less than 2.5%, and more preferably less than 1%; the 17-AAG of Formulations 10, 11, 12, 13, 14, 15, 16, 17, and 18 exhibits the same stability. Furthermore, the 17-AAG of Formulations 1 through 18 — at room temperature, 4 ⁰C, and -20 °C — degrades less than 5% over a period of 14 hours, preferably less than 2.5%, and more preferably less than 1%. [0051] Compared to prior art formulations, the present formulation offers a number of advantages. It is easily prepared and stored, and it avoids the use of excessive amounts of DMSO, which can have poor patient acceptance due to its odor. The present formulation furthermore allows the delivery of a requisite amount of 17-AAG within an acceptable infusion time (e.g., 90 min.).

[0052] The present invention also provides a method for administering 17-AAG to a patient in need thereof, comprising the steps of:

(a) providing a pharmaceutical formulation comprising 17-AAG in a concentration of between about 1 mg/mL and about 5 mg/mL dissolved in a vehicle comprising (i) a first component that is an aprotic, polar solvent in an amount between about 0.1 and about 10 volume %; and, (ii) a second component comprising between about 5.0 and about 55.0 volume % long chain triglycerides in an amount between 90.0 and 99.9 volume %;

(b) including the pharmaceutical formulation into an apparatus that can deliver it to a patient through an intravenous route; and,

(c) administering the formulation intravenously to a patient.

[0053] Examples of 17-AAG formulations administered to patients include, without limitation, Formulations 1 through 18 discussed above. Any suitable apparatus may be used to delivery the formulation intravenously, including an IV bag with attached medical tubing. Finally, the rate of intravenous delivery can be readily determined by one of ordinary skill in the art using well-known methods.

[0054] The present invention further provides a method for preparing a pharmaceutical formulation comprising 17-AAG, comprising the steps of:

(a) providing an amount of 17-AAG;

(b) combining the 17-AAG with a first component, which is an aprotic, polar solvent to provide a solution;

[0055] The amount of 17-AAG provided is typically such that a concentration between about

1.0 mg/mL and 5 mg/mL results in the formulation. Where the first component is DMSO, an amount is oftentimes provided that results in a formulation concentration from about 2.0 mg/mL to about 3.0 mg/mL; where the first component is DMA, an amount is oftentimes provided that results in a formulation concentration from about 1.50 mg/mL to about 3.0 mg/mL.

[0056] For the preparation, the 17-AAG is dissolved in the first component — an aprotic, polar solvent such as DMSO or DMA — to provide a solution. The solution is combined with components that typically include a substantial amount of water, along with long chain triglycerides and phospholipids. Examples of suitable components include, without limitation, Mralipid 10%; Intralipid 20%; Liposyn II 10%; and, Liposyn II 20%. Should some degree of precipitation occur upon combining the solution with the vehicle, the resulting formulation is usually filtered. Examples of formulations prepared through this method include, without limitation, Formulations 1 through 18 discussed above.

[0057] Geldanamycin is a well-known natural product, obtainable by culturing the producing organism, Streptomyces hygroscopicus var. geldanus NRRL 3602. 17-AAG is made semi- synthetically from geldanamycin, by reaction of geldanamycin with allylamine, as described in Sasaki et al, US 4,261,989 (1981), the disclosure of which is incorporated herein by reference.

[0058] 17-AAG administered via a pharmaceutical solution formulation of this invention can be used for treating diseases such as, but not limited to, hyperproliferative diseases, including: cancers of the head and neck which include tumors of the head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, hypopharynx, salivary glands, and paragangliomas; cancers of the liver and biliary tree, particularly hepatocellular carcinoma; intestinal cancers, particularly colorectal cancer; treat ovarian cancer; small cell and non-small cell lung cancer; breast cancer sarcomas, such as fibrosarcoma, malignant fibrous histiocytoma, embryonal rhabdomysocarcoma, leiomysosarcoma, neuioflbrosarcoma, osteosarcoma, synovial sarcoma, liposarcoma, and alveolar soft part sarcoma, neoplasms of the central nervous systems, particularly brain cancer; lymphomas such as Hodgkin's lymphoma, lymphoplasmacytoid lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue lymphoma, mantle cell lymphoma, B-lineage large cell lymphoma, Burkitt's lymphoma, and T-cell anaplastic large cell lymphoma. Clinically, practice of the methods and use of compositions described herein will result in a reduction in the size or number of the cancerous growth and/or a reduction in associated symptoms (where applicable). Pathologically, practice of the method and use of compositions described herein will produce a pathologically relevant response, such as: inhibition of cancer cell proliferation, reduction in the size of the cancer or tumor, prevention of further metastasis, and inhibition of tumor angiogenesis. The method of treating such diseases comprises administering a therapeutically effective amount of an inventive combination to a subject. The method may be repeated as necessary.

[0059] Non-cancer disorders that are characterized by cellular hyperproliferation can also be treated by 17-AAG administered in accordance with this invention. Illustrative examples of such disorders include but are not limited to: atrophic gastritis, inflammatory hemolytic anemia, graft rejection, inflammatory neutropenia, bullous pemphigoid, coeliac disease, demyelinating neuropathies, dermatomyositis, inflammatory bowel disease (ulcerative colitis and Crohn's disease), multiple sclerosis, myocarditis, myositis, nasal polyps, chronic sinusitis, pemphigus vulgaris, primary glomerulonephritis, psoriasis, surgical adhesions, stenosis or restenosis, scleritis, scleroderma, eczema (including atopic dermatitis, irritant dermatitis, allergic dermatitis), periodontal disease ( periodontitis), polycystic kidney disease, and type I diabetes.

[0060] Other examples include vasculitis (e.g., Giant cell arteritis (temporal arteritis, Takayasu's arteritis), polyarteritis nodosa, allergic angiitis and granulomatosis (Churg- Strauss disease), polyangitis overlap syndrome, hypersensitivity vasculitis (Henoch- Schonlein purpura), serum sickness, drug-induced vasculitis, infectious vasculitis, neoplastic vasculitis, vasculitis associated with connective tissue disorders, vasculitis associated with congenital deficiencies of the complement system, Wegener's granulomatosis, Kawasaki's disease, vasculitis of the central nervous system, Buerger's disease and systemic sclerosis); gastrointestinal tract diseases (e.g., pancreatitis, Crohn's disease, ulcerative colitis, ulcerative proctitis, primary sclerosing cholangitis, benign strictures of any cause including ideopathic (e.g., strictures of bile ducts, esophagus, duodenum, small bowel or colon); respiratory tract diseases (e.g., asthma, hypersensitivity pneumonitis, asbestosis, silicosis and other forms of pneumoconiosis, chronic bronchitis and chronic obstructive airway disease); nasolacrimal duct diseases (e.g., strictures of all causes including ideopathic); and eustachean tube diseases (e.g., strictures of all causes including ideopathic).

[0061] 17-AAG can be administered in combination with other anti-cancer or cytotoxic agents, including alkylating agents, angiogenesis inhibitors, antimetabolites, DNA cleavers, DNA crosslinkers, DNA intercalators, DNA minor groove binders, heat shock protein 90 inhibitors, histone deacetylase inhibitors, microtubule stabilizers, nucleoside (purine or pyrimidine) analogs, proteasome inhibitors, topoisomerase (I or II) inhibitors, tyrosine kinase inhibitors. Specific anti-cancer or cytotoxic agents include β-lapachone, 17- DMAG, bicalutamide, bleomycin, bortezomib, bisulfan, calicheamycin, camptothecin, capecitabine, callistatin A, CC-1065, cisplatin, cryptophycins, daunorubicin, discodermolide, docetaxel, doxorubicin, duocarmycin, dynemycin A, epothilones, etoposide, floxuridine, fludarabine, fluoruracil, gefitinib, geldanamycin, gemcitabine, hydroxyurea, imatinib, interferons, interleukins, irinotecan, leptomycin B, methotrexate, mitomycin C, oxaliplatin, paclitaxel, spongistatins, suberoylanilide hydroxamic acid (SAHA), thiotepa, topotecan, trichostatin A, vinblastin, vincristine, and vindesine.

[0062] The co-administered anti-cancer or cytotoxic agent can be a protein kinase inhibitor, including: quinazolines, particularly 4-anilinoquinazolines such as Iressa (AstraZeneca; N- (3- chloro-4-fluorophenyl)-7-methoxy-6-[3(4- quinazolinamine) and Tarceva (Roche/Genentech; N-(3-ethynylphenyl)-6,7- bis(2-methoxyethoxy)-4-quinazolinamine monohydrochloride); phenylamino-pyrimidines such as Gleevec (Novartis; 4-[(4-methyl-l- piperazinyl)methyl]-N-[4-methyl-3-[[4-(3-pyridinyl)-2- pyrimidinyl]amino]phenyl]benzamide); pyrrolo- and pyrazolopyrimidines such as BEBX 1382 (Boehringer Ingelheim; N8-(3-chloro-4-fluorophenyl)-N-2-(l-rnethyl-4-piperidinyl)- pyrimido[5,4-d] pyrimidine-2,8-diamine); indoles and oxindoles such as Semaxinib (Pharmacia; 3-[(3,5-dimethyl-l H-pyrrol-2-yl)methylene]-l,3-dihydro-2H- Indol-2-one); benzylidene malononitriles; flavones such as flavopiridol (Aventis; 2-(2-chlorophenyl)-5 ,7- dihydroxy-8-[(3S,4R)-3-hydroxy-l-methyl- 4-piperidinyl]-4H-l -benzopyran-4-one); staurosporines such as CEP-701 (Cephalon); antibodies such as Herceptin (Genentech); and, ribozyrnes such as Angiozyme (Ribozyme Pharmaceuticals).

[0063] Using a pharmaceutical solution formulation of this invention, 17-AAG may be administered in a dose ranging from about 4 mg/m² to about 4000 mg/m² depending on the frequency of administration. A preferred dosage regimen for 17-AAG is about 450 mg/m² weekly (Banerji et al, Proc. Am, Soc. Clin. Oncol. , 22, 199 (2003, abstract 797), "A Pharmacokinetically (PK)-pharmacodynamically (PD) Guided Phase I Trial of the Heat Shock Protein 90 (HSP90) Inhibitor 17-Allyl-17-demethoxygeldanamycin (17AAG)"). Alternatively, a dose of about 308 mg/m² weekly can be administered. See Goetz et al, Eur. J. Cancer, 38 (Supp. 7), S54-S55 (2002), "A phase I trial of 17-Allyl-Amino-Geldanamycin (17-AAG) in patients with advanced cancer."

[0064] The practice of this invention can be further understood by reference to the following examples, which are provided by way of illustration and not of limitation.

General Experimental Section

[0065] Instruments and reagents used for the experiments described below were obtained from the indicated vendor, where applicable: water bath (VWR Polyscience Model 1167); balance (Mettler-Toledo AT261 Delta Range); centrifuge (Sorval Super T21); HPLC (Agilent 1100 Series); reverse-phase column (Agilent ZORBAX Eclipse® C8 Column); guard column (Agilent ZORBAX Eclipse® XDB-C8 Column); osmometer (Wescor Vapro™ Vapor Pressure Osmometer); dimethylacetamide (Sigma); dimethyl sulfoxide (J.T. Baker); Safflower Oil (commercial); soybean oil (commercial); sesame oil (commercial); and Miglyol 810 (Sasol North America).

Example 1

Solubility of 17-AAG in Various Oils

[0066] 1 mg of 17-AAG reference standard was initially dissolved in 1 mL of 100% methanol. The sample was diluted to 0.4 mg/mL concentration with methanol and used as a reference standard. 17-AAG was dissolved in various organic solvents or oils. The formulations were lightly vortexed to ensure homogeneity and subsequently filtered through 0.2 μm PVDF syringe filters. The 17-AAG stock formulations were diluted in their respective solvents to determine the initial concentration and purity as well as the recovery of 17-AAG by RP-HPLC analysis. [0067] 20 mg of 17-AAG was taken in different vials and 0.2 niL of various oils was added as listed in Table 1. The samples were vortexed to dissolve 17-AAG. Upon preparation, the samples showed varying degrees of precipitation and were subsequently filtered through 0.2 μm PVDF filters and analyzed by RP-HPLC.

Table 1: Visual appearance, solubility and purity of 17-AAG/oil formulations.

Example 2

Evaluation of DMA in Combination with Oils on 17-AAG Solubility [0068] 17-AAG stocks were prepared in 100% DMA (50 mg/mL or 120 mg/mL). The formulations listed in Tables 2 to 5 were prepared by two different methods. Either the 17- AAG/solvent stock was slowly added to excess oil (Method I), or the oil was added to an aliquot of the 17-AAG/solvent stock (Method II). For example, solvent to oil (Method I) was prepared as follows: A 20 μL or 50 mg/mL 17-AAG/DMA stock was added to vial containing 500 μL of the Safflower oil under stiπing conditions. Oil to solvent (Method II) was prepared by adding 500 μL of oil dropwise to a vial containing 20 μL of 50 mg/mL 17- AAG/DMA stock over a period of 5 minutes. The sample was stirred on a magnetic stir plate for a period of 30 minutes. Upon preparation, the DMA samples showed varying degrees of precipitation. The formulations were subsequently filtered using 0.2 μm PVDF filters and analyzed by RP-HPLC. There was no change in appearance observed in any of the formulations tested after 24 hours of incubation at room temperature. [0069] Table 2 provides data regarding an evaluation of 17-AAG solubility using DMA in combination with various oils. The combinations were prepared in accordance with Method I where a 50 mg/mL stock solution of 17-AAG/DMA was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (2 mg/mL). Table 2: Evaluation of DMA in combination with oils on 17- AAG solubility. RP-HPLC analysis (17-AAG/DMA stock: 50 mg/mL), Method I.

[0070] Table 3 provides data regarding an evaluation of 17-AAG solubility using DMA in combination with various oils. The combinations were prepared in accordance with Method II, where a 50 mg/mL stock solution of 17-AAG/DMA was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (2 mg/mL).

Table 3: Evaluation of DMA in combination with oils on 17-AAG solubility. RP-HPLC analysis (17-AAG/DMA stock: 50 mg/mL), Method II.

[0071] Table 4 provides data regarding an evaluation of 17-AAG solubility using DMA in combination with various oils. The combinations were prepared in accordance with Method I, where a 120 mg/mL stock solution of 17-AAG/DMA was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (4.8 mg/mL).

Table 4: Evaluation of combination of DMA with oils on 17-AAG solubility (17- AAG/DMA stock: 120 mg/mL), Method I.

[0072] Table 5 provides data regarding an evaluation of 17- AAG solubility using DMA in combination with various oils. The combinations were prepared in accordance with Method II where a 120 mg/mL stock solution of 17-AAG/DMA was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (4.8 mg/mL).

Table 5: Evaluation of DMA in combination with oils on 17-AAG solubility (17- AAG/DMA stock: 120 mg/mL), Method II.

Example 3

Evaluation of DMSO in Combination with Oils on 17-AAG Solubility [0073] 17-AAG stocks were prepared in 100% DMSO (50 mg/mL or 200 mg/mL). The formulations listed below in Tables 6 to 9 were prepared by two different methods. Either the 17-AAG solvent stock was slowly added to excess oil (Method I), or the oil was added to an aliquot of the 17-AAG/solvent stock (Method II). For example, solvent to oil (Method I) was prepared as follows: A 20 μL of 50 mg/mL 17-AAG/DMSO stock was added to a vial containing 500 μL of the Safflower oil under stirring conditions. Oil to solvent (Method II) was prepared by adding 500 μL of oils dropwise to a vial containing 20 μL of 50 mg/mL 17- AAG/DMSO stock over a period of 5 minutes. The sample was stirred on a magnetic stir plate for a period of 30 minutes. Upon preparation, the formulations formed tiny oil droplets initially and precipitated after 10 minutes. The formulations were subsequently filtered using 0.2 μm PVDF filters and analyzed by RP-HPLC. The DMSO containing formulations showed oil droplet formation even after filtering; thus supernatant without the oil droplets was loaded on the RP-HPLC. There was no change in the appearance observed in any of the formulations tested after 24 hours of incubation at room temperature. [0074] Table 6 provides data regarding an evaluation of 17-AAG solubility using DMSO in combination with various oils. The combinations were prepared in accordance with Method I, where a 50 mg/mL stock solution of 17-AAG/DMSO was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (2 mg/mL).

Table 6: Evaluation of DMSO in combination with oils on 17-AAG solubility (17- AAG/DMSO stock: 50 mg/mL), Method I.

[0075] Table 7 provides data regarding an evaluation of 17-AAG solubility using DMSO in combination with various oils. The combinations were prepared in accordance with Method π, where a 50 mg/mL stock solution of 17-AAG/DMSO was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (2 mg/mL).

Table 7: Evaluation of DMSO in combination with oils on 17-AAG solubility (17- AAG/DMSO stock: 50 mg/mL), Method II.

[0076] Table 8 provides data regarding an evaluation of 17-AAG solubility using DMSO in combination with various oils. The combinations were prepared in accordance with Method I, where a 200 mg/mL stock solution of 17-AAG/DMSO was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (8 mg/mL).

Table 8: Evaluation of DMSO in combination with oils on 17-AAG solubility (17- AAG/DMSO stock: 200 mg/mL), Method I.

[0077] Table 9 provides data regarding an evaluation of 17-AAG solubility using DMSO in combination with various oils. The combinations were prepared in accordance with Method II, where a 200 mg/mL stock solution of 17-AAG/DMSO was used. Percent recovery was calculated by dividing the solubility of 17-AAG by the targeted concentration (8 mg/mL).

Table 9: Evaluation of DMSO in combination with oils on 17-AAG solubility (17- AAG/DMSO stock: 200 mg/mL), Method II.

Example 4

Evaluation of Combination of Solvents and Oils on 17-AAG Solubility [0078] 17-AAG stocks were prepared in either 100% DMA (50 mg/mL or 120 mg/mL) or DMSO (50 mg/mL or 200 mg/mL). The formulations were prepared as follows: A 10 or 20 μL aliquot of 17-AAG/solvent stock was added to a vial containing 500 μL of the oil and gently swirled for 20 seconds to ensure complete homogeneity. The samples were subsequently incubated at room temperature and monitored over the course of 2 hours. [0079] Table 10 presents the visual appearance and solubility profiles of 17-AAG formulations containing a combination of DMA and oils prepared with no mechanical stress. Upon preparation, the DMA samples showed either little or no evidence of precipitation.

Table 10: Evaluation of Combination of Solvents and Oils on 17-AAG Solubility

[0080] Table 11 presents the visual appearance and solubility profiles of 17-AAG formulations containing combinations of DMSO and oils prepared with no mechanical shearing. Upon preparation, the DMSO samples showed tiny oil droplets. After 2 hours of incubation at room temperature, there was a large amount of particles settled on the bottom of the vial containing DMSO/oil combinations in comparison to the DMA/oil formulations, irrespective of the stock concentrations used.

Table 11: Evaluation of Combination of Solvents and Oils on 17-AAG Solubility

Example 5

17-AAG Solubility and Stability in 10% Intralipid

[0081] Different stock concentrations of 17-AAG were prepared either in 100% DMA or DMSO as follows: 17-AAG/DMA— 50, 55, 60, and 65 mg/mL; 17-AAG/DMSO— 50, 55, 60, and 65 mg/mL.

[0082] The formulations listed below in Tables 12 to 15 were prepared by mixing different aliquots of each 17-AAG/solvent stock in 1 mL of 10% Intralipid. For example, a 50/25 of 17-AAG/DMA/10% Intralipid combination was prepared as follows: 25 μL of 50 mg/ML 17-AAG/ DMA stock was slowly added to vial containing 1 mL of the 10% Intralipid. The sample was mixed by inverting the vials continuously for 20 seconds. All of the formulations were incubated at room temperature for a period of 24 hours to monitor the stability of 17- AAG in the lipid emulsions.

[0083] Table 12 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 50 mg/mL 17-AAG stock and formulated with 10% Intralipid.

[0084] Table 13 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 55 mg/rnL 17-AAG stock and formulated with 10% Intralipid.

Table 13: Stability of 17-AAG in 10% Intralipid by visual appearance.

[0085] Table 14 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 60 mg/mL 17-AAG stock and formulated with 10% Intralipid.

Table 14: Stability of 17-AAG in 10% Intralipid by visual appearance.

[0086] Table 15 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 65 mg/mL 17-AAG stock and formulated with 10% Intralipid.

Table 15: Stability of 17-AAG in 10% Intralipid by visual appearance.

particulates particulates particulates particulates

Example 6

17 -AAG Solubility and Stability in 10% Liposyn 2

[0087] Different stock concentrations of 17-AAG were prepared either in 100% DMA or DMSO as follows: 17-AAG/DMA— 55, 60, and 65 mg/mL; 17-AAG/DMSO— 55, 60 and 65 mg/mL. The formulations listed below in Tables 16 to 18 were prepared by mixing different aliquots of each 17-AAG/solvent stock in 1 mL of 10% Liposyn II. For example, a 55/25 of 17-AAG/DMA/10% Liposyn II combination was prepared as follows: 25 μL of 55 mg/mL 17-AAG/DMA stock was slowly added to a vial containing 1 mL of the 10% Liposyn II. The sample was mixed by inverting the vials continuously for 20 seconds. AU of the formulations were incubated at room temperature for a period of 24 hours to monitor the stability of 17-AAG in the lipid emulsions.

[0088] Table 16 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 55 mg/mL 17-AAG stock and formulated with 10% Liposyn II.

Table 16: Stability of 17-AAG in 10% Liposyn II by visual appearance.

[0089] Table 17 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 60 mg/mL 17-AAG stock and formulated with 10% Liposyn II.

Table 17: Stability of 17-AAG in 10% Liposyn II by visual appearance.

[0090] Table 18 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 65 mg/mL 17-AAG stock and formulated with 10% Liposyn II.

Table 18: Stability of 17-AAG in 10% Liposyn II by visual appearance.

Example 7

17-AAG Solubility and Stability in 10% or 20% Intralipid

[0091] Different stock concentrations of 17-AAG were prepared either in 100% DMA or DMSO as follows: 17-AAG/DMA— 70, 80, 90, 95 and 100 mg/mL; 17-AAG/DMSO— 70, 80 90, 95 and 100 mg/mL. The formulations listed below in Tables 19 to 20 were prepared by mixing different aliquots of each 17-AAG/solvent stock in 1 mL of 10 or 20% Intralipid. For example, a 70/25 of 17-AAG/DMA/Intralipid combination was prepared as follows: 25 μL of 70 mg/mL 17-AAG/DMA stock was slowly added to a vial containing 1 mL of Intralipid. The sample was mixed by inverting the vials continuously for 40 seconds. AU of the formulations were incubated at room temperature for a period of 24 hours to monitor the stability of 17-AAG in the lipid emulsions.

[0092] Table 19 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 70 mg/mL 17-AAG stock and formulated with 10% Intralipid.

Table 19: Stability of 17-AAG in 10% Intralipid by visual appearance following dilution in 10% Intralipid.

[0093] Table 20 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 70-80 mg/mL 17-AAG stock and formulated with 20% Intralipid.

Table 20: Stability of 17-AAG in 20% Intralipid by visual appearance.

Example 8

17-AAG Solubility and Stability in 10% or 20% Liposyn II

[0094] Different stock concentrations of 17-AAG were prepared either in 100% DMA or DMSO as follows: 17-AAG/DMA— 70, 80, 90, 95 and 100 mg/mL; 17-AAG/DMSO— 70, 80, 90, 95 and 100 mg/mL. The formulations listed below in Tables 21 to 23 were prepared by mixing different aliquots of each 17-AAG/solvent stock in 1 mL of 10 or 20% Liposyn II. For example, a 70/25 of 17-AAG/DMA/Liposyn II combination was prepared as follows: 25 μL of 70 mg/mL 17-AAG/DMA stock was slowly added to vial containing 1 mL of Liposyn π. The sample was mixed by inverting the vials continuously for 40 seconds. All of the formulations were incubated at room temperature for a period of 24 hours to monitor the stability of 17-AAG in the lipid emulsions.

[0095] Table 21 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 70 or 80 mg/mL 17-AAG stocks and 10% Liposyn II.

Table 21: Stability of 17-AAG in 10% Liposyn II by visual appearance.

[0096] Table 22 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 70-80 mg/rnL 17-AAG stocks and 20% Liposyn II.

Table 22: Stability of 17-AAG in 20% Liposyn II by visual appearance following dilution in 20% Liposyn II.

[0097] Table 23 below presents the visual observation for 17-AAG/DMA or DMSO emulsions prepared from a 90-100 mg/mL 17-AAG stock and 20% Liposyn II.

Table 23: Stability of 17-AAG in 20% Liposyn II by visual appearance.

Example 9

Stability of 17 -AAG Lipid Emulsions

[0098] Emulsion particle size was measured after adding 17-AAG to the fat emulsions using a light microscope. 10 μL aliquots of 17-AAG emulsions were loaded onto a RP-HPLC. Osmolality was determined by removing 10 μL aliquots of the 17-AAG emulsions and measuring the quantity using a Wescor Vapro™ Vapor Pressure Osmometer. [0099] Table 24 below presents the microscopic observations of lipid emulsions before and after the addition of 17-AAG. Using a light microscope, the oil droplet of the emulsion was categorized into 3 different sizes: small (< 200 nm); medium (200-400 nm) or large droplets (> 400 nm). Table 24: Stability of 17-AAG in lipid emulsions by microscopic observation.

[00100] Table 25 below presents the stability of 17-AAG/DMA or DMSO emulsions prepared from a 55, 70 or 100 mg/mL 17-AAG stocks and 20% Liposyn II.

Table 25: Stability of 17-AAG in 20% Liposyn II by visual appearance following dilution in 20% Liposyn II.

[00101 ] Table 26 below presents the osmolality of the commercially available lipid emulsions with no drug.

Table 26: Osmolality of commercially available lipid emulsions with no drug.

Table 27: Osmolality of 17-AAG/DMSO emulsions.

Table 28: Osmolality of 17-AAG/DMA emulsions.

Example 10

17 -AAG Emulsions

[00102] Tables 29 and 30 below present various 17-AAG formulations/emulsions with the final dosage/ volume of active and inactive ingredients to be infused.

Table 29: Final 17-AAG and DMSO concentrations in various 17-AAG formulations/ emulsions in fat emulsions.

Table 30: Final 17-AAG and DMA concentrations in various 17-AAG formulations/ emulsions in fat emulsions

Example 11

Short-Term Stability of 17 -AAG Formulations

[00103] A 100 mg/mL stock of 17-AAG was prepared either in 100% DMA or

DMSO. Aliquots of 17-AAG/solvent formulations were placed in glass vials, covered with aluminum foil and incubated at -20 ⁰C, 4 ⁰C or room temperature {i.e., RT) for a period up to 28 days. Samples were withdrawn at each time point (day 7, 14, 21 or 28), and an aliquot from each sample was subsequently diluted in and analyzed by RP-HPLC to determine the percent recovery and purity of the stock solutions. The stability of 17-AAG/DMA or DMSO test samples was also examined by diluting the stock samples in 20% Liposyn II. The 17- AAG emulsions were incubated at room temperature over a period of 24 hours to determine percent recovery via RP-HPLC.

[00104] Table 31 below presents the 28 day stability data of 17-AAG stock in either DMA or DMSO at 100 mg/mL 17-AAG concentration incubated at 4 ⁰C, room temperature, or -20 ⁰C. Percent recovery and purity were analyzed after time = 0, 7 hours, 14 hours, 21 hours, and 28 hours. The percent recovery was calculated by dividing the AUP of the tη through t₂₈ formulations by the AUP of to formulations.

Table 31: Short-term stability on 17-AAG formulations by RP-HPLC analysis.

[00105] Table 32 below presents 24 hour stability data of 17-AAG stock in either DMA or DMSO at 100 mg/mL 17-AAG concentration incubated at 4 ⁰C, room temperature, or -20 ⁰C.

Table 32: 17-AAG Lipid Emulsion Stability after 24 hours of incubation at RT in 20% Liposyn II.

[00106] Table 33 below presents the percent recovery of t₂₁ or t₂₈ 17-

AAG/DMA or DMSO lipid emulsions in 20% Liposyn after 24 hours of incubation at room temperature.

Table 33: 17-AAG Lipid Emulsion Stability after 24 hours of incubation at RT in 20% Liposyn II.

[00107] The foregoing detailed description of the invention includes passages that are chiefly or exclusively concerned with particular parts or aspects of the invention. It is to be understood that this is for clarity and convenience, that a particular feature may be relevant in more than just the passage in which it is disclosed, and that the disclosure herein includes all the appropriate combinations of information found in the different passages. Similarly, although the various descriptions herein relate to specific embodiments of the invention, it is to be understood that where a specific feature is disclosed in the context of a particular embodiment, such feature can also be used, to the extent appropriate, in the context of another embodiment, in combination with another feature, or in the invention in general.

Claims

CLAIMS OF THE INVENTIONWhat is claimed is:

1. A pharmaceutical formulation comprising 17-AAG in an amount from about 1.0 mg/mL to about 5.0 mg/mL dissolved in a vehicle comprising (i) a first component that is an aprotic, polar solvent in an amount between about 0.1 and about 10 volume %; and, (ii) a second component that is an aqueous mixture comprising between about 5.0 and about 55.0 volume % long chain triglycerides, in an amount between about 90.0 and 99.9 volume %.

2. The pharmaceutical formulation according to claim 1, wherein the aprotic, polar solvent is dimethylsulfoxide, N,N-dimethylacetamide or combinations thereof.

3. The pharmaceutical formulation according to claim 1, wherein the fatty acids of the long chain triglycerides of the second component comprise linoleic acid, oleic acid and palmitic acid.

4. The pharmaceutical formulation according to claim 1, wherein the formulation further comprises phospholipids, and wherein the phospholipids are egg phospholipids present in an amount between about 0.8 and 3.0 volume percent.

5. The pharmaceutical formulation according to claim 2, wherein 17-AAG is present in the formulation in an amount between about 1.50 mg/mL and 3.0 mg/mL.

6. The pharmaceutical formulation according to claim 5, wherein the fatty acids of the long chain triglycerides comprise about 50 weight % to about 70 weight % linoleic acid, about 15 weight % to about 25 weight % oleic acid and about 7 weight % to about 10 weight % palmitic acid.

7. The pharmaceutical formulation according to claim 6, wherein the concentration of the aprotic, polar solvent ranges from about 1.0 % to about 4.0 % by volume.

8. The pharmaceutical formulation according to claim 7, wherein the formulation further comprises glycerin, and wherein the glycerin is present in an amount between about 2.0 and about 3.0 volume %. 9. A method for administering 17-AAG to a patient in need thereof, comprising the steps of:

(a) providing a pharmaceutical formulation comprising 17-AAG in a concentration of between about 1.0 mg/mL and about 5.0 mg/mL dissolved in a vehicle comprising (i) a first component that is an aprotic, polar solvent in an amount between about 0.1 and about 10 volume %; and, (ii) a second component that is an aqueous mixture comprising between about 5.0 and about 55.0 volume % long chain triglycerides, in an amount between about 90.0 and 99.

9 volume %;

(c) administering the formulation intravenously to a patient.

10. The method according to claim 9, wherein the aprotic, polar solvent is dimethylsulfoxide, N,N-dimethylacetamide or combinations thereof.

11. The method according to claim 9, wherein fatty acids of the long chain triglycerides of the second component comprise linoleic acid, oleic acid and palmitic acid.

12. The method according to claim 9, wherein the vehicle further comprises phospholipids, and wherein the phospholipids are egg phospholipids present in an amount between about 0.8 and 3.0 volume percent.

13. The method according to claim 10, wherein the 17-AAG is present in the formulation in an amount between about 1.50 mg/mL and 3.0 mg/mL.

14. The method according to claim 13, wherein the fatty acids of the long chain triglycerides comprise about 50% to about 70% linoleic acid, about 15% to about 25% oleic acid and about 7% to about 10% palmitic acid.

15. The method according to claim 14, wherein the concentration of the aprotic, polar solvent ranges from about 1.0 % to about 4.0 % by volume.

16. The method according to claim 15, wherein the formulation further comprises glycerin in an amount between about 2.0 and about 3.0 volume %.

17. The method according to claim 16, wherein the 17-AAG is administered in an amount from about 4 mg/m² to about 4000 mg/m².

18. The method according to claim 16, wherein the 17-AAG is administered in an amount of about 450 mg/m² weeldy.

19. The method according to claim 16, wherein the 17-AAG is administered in an amount of about 308 mg/m² weekly.

20. A method for preparing a pharmaceutical formulation comprising 17-AAG, comprising the steps of:

(a) providing an amount of 17-AAG;

21. The method according to claim 20, wherein the aprotic, polar solvent is dimethylsulfoxide, N,N-dimethylacetamide or combinations thereof.

22. The method according to claim 20, wherein the fatty acids of the long chain triglycerides of the second component comprise linoleic acid, oleic acid and palmitic acid.

23. The method according to claim 20, wherein the vehicle further comprises phospholipids, and wherein the phospholipids are egg phospholipids present in an amount between about 0.8 and 3.0 volume percent.

24. The method according to claim 21, wherein 17-AAG is provided such that its concentration in the formulation is between about 1.50 mg/mL and 3.0 mg/mL.

25. The method according to claim 24, wherein the fatty acids of the long chain triglycerides comprise about 50% to about 70% linoleic acid, about 15% to about 25% oleic acid, and about 7% to about 10% palmitic acid.

26. The method according to claim 25, wherein aprotic, polar solvent is provided such that its concentration in the formulation ranges from about 1.0 % to about 4.0 % by volume.

27. The method according to claim 26, wherein the vehicle further comprises glycerin in an amount between about 2.0 and about 3.0 volume %.