PCT
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`WO 00/06152
`
`WORLD INTELLECTUAL PROPERTY ORGANIZATION
`International Bureau
`
`(51) International Patent Classification 7 :
`A61K 31/337, 47/48, 47/42
`
`Al
`
`(11) International Publication Number:
`
`(43) International Publication Date:
`
`10 Febrnary 2000 ( 10.02.00)
`
`(21) International Application Number:
`
`PCT/US99/17179
`
`(22) International Filing Date:
`
`29 July 1999 (29.07.99)
`
`(30) Priority Data:
`60/094,687
`
`30 July 1998 (30.07.98)
`
`us
`
`(71) Applicant
`except US):
`States
`designated
`all
`(for
`NOVOPHARM BIOTECH, INC. [CA/CA]; 30 Novopharm
`Court, Toronto, Ontario MIB 2K9 (CA).
`
`(72) Inventors; and
`(75) Inventors/Applicants (for US only): KADIMA, Tenshuk, A.
`[CA/CA]; 7 Woodington Bay, Winnipeg, Manitoba R3P
`1M6 (CA). KAPLAN, Howard, A. [CA/CA]; 18 Hillhouse
`Road, Winnipeg, Manitoba R2V 2V9 (CA). TUTTLE,
`Robert, C. [US/CA]; 782 Allegheny Drive, Winnipeg,
`Manitoba R3T 5L2 (CA).
`
`(74) Agents: WU, Frank et al.; Morrison & Foerster LLP, 755 Page
`Mill Road, Palo Alto, CA 94304-1018 (US).
`
`(81) Designated States: AE, AL, AM, AT, AU, AZ, BA, BB, BG,
`BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB,
`GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP,
`KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK,
`MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI,
`SK, SL, TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU, ZA,
`ZW, ARIPO patent (GH, GM, KE, LS, MW, SD, SL, SZ,
`UG, ZW), Eurasian patent (AM, AZ, BY, KG, KZ, MD,
`RU, TJ, TM), European patent (AT, BE, CH, CY, DE, DK,
`ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OAPI
`patent (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR,
`NE, SN, TD, TG).
`
`Published
`With international search report.
`Before the expiration of the time limit for amending the
`claims and to be republished in the event of the receipt of
`amendments.
`
`(54) Title: PHARMACEUTICALLY ACCEPTABLE COMPOSITION COMPRISING AN AQUEOUS SOLUTION OF PACLITAXEL
`AND ALBUMIN
`
`(57) Abstract
`
`An optically clear, pharmaceutically acceptable aqueous composition comprising paclitaxel or a derivative thereof, sernm albumin
`and a pharmaceutically acceptable vehicle, wherein the composition comprises no more than 10 % organic solvent and has a pH of about
`3.0 to about 4.8, is described. The sernm albumin can be fatted or defatted, and the composition can optionally be lyophilized or optionally
`lyophilized and reconstituted. At least 70 % of the paclitaxel is bound to sernm albumin, the ratio of paclitaxel to albumin is at least about
`I :5, and the concentration of paclitaxel is at least about 25 µg!ml. Methods of making and using this composition are also provided.
`
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`

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`FOR THE PURPOSES OF INFORMATION ONLY
`
`Codes used to identify States party to the PCT on the front pages of pamphlets publishing international applications under the PCT.
`
`AL
`AM
`AT
`AU
`AZ
`BA
`BB
`BE
`BF
`BG
`BJ
`BR
`BY
`CA
`CF
`CG
`CH
`CI
`CM
`CN
`cu
`CZ
`DE
`DK
`EE
`
`Albania
`Armenia
`Austria
`Australia
`Azerbaijan
`Bosnia and Herzegovina
`Barbados
`Belgium
`Burkina Faso
`Bulgaria
`Benin
`Brazil
`Belarus
`Canada
`Central African Republic
`Congo
`Switzerland
`Ci\te d'Ivoire
`Cameroon
`China
`Cuba
`Czech Republic
`Germany
`Denmark
`Estonia
`
`ES
`FI
`FR
`GA
`GB
`GE
`GH
`GN
`GR
`HU
`IE
`IL
`IS
`IT
`JP
`KE
`KG
`KP
`
`KR
`KZ
`LC
`LI
`LK
`LR
`
`Spain
`Finland
`France
`Gabon
`United Kingdom
`Georgia
`Ghana
`Guinea
`Greece
`Hungary
`Ireland
`Israel
`Iceland
`Italy
`Japan
`Kenya
`Kyrgyzstan
`Democratic People's
`Republic of Korea
`Republic of Korea
`Kazakstan
`Saint Lucia
`Liechtenstein
`Sri Lanka
`Liberia
`
`LS
`LT
`LU
`LV
`MC
`MD
`MG
`MK
`
`ML
`MN
`MR
`MW
`MX
`NE
`NL
`NO
`NZ
`PL
`PT
`RO
`RU
`SD
`SE
`SG
`
`Lesotho
`Lithuania
`Luxembourg
`Latvia
`Monaco
`Republic of Moldova
`Madagascar
`The former Yugoslav
`Republic of Macedonia
`Mali
`Mongolia
`Mauritania
`Malawi
`Mexico
`Niger
`Netherlands
`Norway
`New Zealand
`Poland
`Portugal
`Romania
`Russian Federation
`Sudan
`Sweden
`Singapore
`
`SI
`SK
`SN
`sz
`TD
`TG
`TJ
`TM
`TR
`TT
`UA
`VG
`us
`uz
`VN
`YU
`zw
`
`Slovenia
`Slovakia
`Senegal
`Swaziland
`Chad
`Togo
`Tajikistan
`Turkmenistan
`Turkey
`Trinidad and Tobago
`Ukraine
`Uganda
`United States of America
`Uzbekistan
`Viet Nam
`Yugoslavia
`Zimbabwe
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`WO 00/06152
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`PCT/US99/17179
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`PHARMACEUTICALLY ACCEPTABLE COMPOSITION COMPRISING AN AQUEOUS SOLUTION OF PACLITAXEL
`AND ALBUMIN
`
`5
`
`10
`
`CROSS-REFERENCE TO RELATED APPLICATIONS
`
`(Not Applicable)
`
`STATEMENT OF RIGHTS TO INVENTIONS
`
`MADE UNDER FEDERALLY-SPONSORED RESEARCH
`
`(Not Applicable)
`
`TECHNICAL FIELD
`
`The present invention relates generally to aqueous formulations of paclitaxel and
`
`methods of use thereof. More specifically, it pertains to pharmaceutical compositions
`
`15
`
`comprising paclitaxel (Ptx) or a derivative thereof and serum albumin or a fragment
`
`thereof, particularly human serum albumin, and more particularly recombinant human
`
`serum albumin, and a physiologically acceptable vehicle; methods of preparation of such
`
`pharmaceutical compositions; and methods of use thereof. The vehicle can comprise an
`
`organic solvent, and the composition lacks a toxic emulsifier such as Cremophor EL®
`
`20
`
`(polyoxyethylated castor oil).
`
`BACKGROUND OF THE INVENTION
`
`Paclitaxel, a structurally complex natural plant product, has demonstrated efficacy
`
`25
`
`in the treatment of a wide variety of human malignancies. This drug shows strong
`
`cytotoxicity in KB cell structures and in several of the National Cancer Institute's in vivo
`
`screens, including the P-388, L-1210, and P-1534 mouse leukemias, the B-16
`
`melanocarcinoma, the CX-1 colon xenograft, the LX-1 lung xenograft, and the MX-1
`
`breast xenograft. Further, studies by McGuire et al. [(1989) Ann. Int. Med. 111 :273-279]
`
`30
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`found paclitaxel to be active against drug-refractory ovarian cancer. Positive results were
`
`also seen with paclitaxel treatment of patients with other cancers, including melanoma.
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`Einsig et al. (1988) Proc. Am. Soc. Clin. Oneal. 7:249; Holmes (1991) J Natl. Cancer Inst.
`
`83:1797-1805; and Kohn et al. (1994) J Natl. Cancer Inst. 86:18-24.
`
`In addition to various cancers, paclitaxel has been used in treating several other
`
`diseases, including malaria and babesiosis. U.S. Patent Nos. 5,356,927 and 5,631,278.
`
`5
`
`Paclitaxel can be used to treat indications characterized by chronic inflammation such as
`
`rheumatoid arthritis and auto-immune disease. U.S. Patent No. 5,583,153; and Song et al.
`
`(1996) Arthritis Rheum. 39:S 178. Paclitaxel can impair chronic inflammation by inhibiting
`
`the activity of white blood cells involved in the inflammatory response; reducing the
`
`production of matrix metalloproteinases that permanently damage tissues; blocking the
`
`10
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`cancer-like growth of previously normal cells which respond to chronic inflammation by
`
`proliferating; and inhibiting the growth of blood vessels which lead to the formation of scar
`
`tissue. Paclitaxel is also a potent inhibitor of angiogenesis and other processes involved in
`
`the development of chronic inflammation. This activity is due, in part, to paclitaxel's
`
`ability to inhibit the transcription factor AP-1. AP-1 is a key regulator of genes involved in
`
`15
`
`the production of (i) matrix metalloproteinases, (ii) cytokines associated with chronic
`
`inflammation, and (iii) proteins necessary for cell proliferation. Therefore, paclitaxel
`
`inhibits a regulator which plays an important role in chronic inflammation and conditions
`
`that are dependent on angiogenesis (new blood vessel formation), including tumor growth.
`
`Paclitaxel has shown strong anti-angiogenic activity when tested in the chorioallantoic
`
`20
`
`membrane of the developing chick embryo. The drug is a more potent angiogenesis
`
`inhibitor than approved anti-arthritic agents such as methotrexate, penicillamine, and
`
`steroids.
`
`Atherosclerosis and restenosis have also been treated with low paclitaxel dosages.
`
`U.S. Patent No. 5,616,608. Paclitaxel can alter several aspects of the process leading to
`
`25
`
`restenosis, including inhibition of vascular smooth muscle cell ("VSMC") migration,
`
`inhibition of VSMC proliferation, and inhibition of the effects of certain growth factors on
`
`these cells. Paclitaxel also inhibits synoviocyte proliferation. Paclitaxel is capable of
`
`inhibiting proliferation of synoviocytes in vitro and inducing apoptosis (programmed cell
`death) at concentrations as low as 10-7 M, and is cytotoxic to the synoviocytes at slightly
`higher concentrations of 1 o-6 to 10-5 M. Paclitaxel inhibits collagenase production by
`chondrocytes in vitro, but is not toxic to normal chondrocytes. A concentration of 10-7 M
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`30
`
`paclitaxel, for example, reduced collagenase expression by over 50% in cultured
`
`2
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`chondrocytes stimulated by tumor necrosis factor and interleukin- I. This inhibition occurs
`
`downstream from the transcription factor activity of c-fos and c-jun, apparently by
`
`disrupting the normal functioning of the AP- I molecule, resulting in inhibition of
`
`transcription of the collagenase gene. As such, inhibition of collagenase secretion by
`
`5
`
`paclitaxel is not strictly due to interruption of the protein secretory pathway, which is
`
`dependent upon microtubule function for the movement of secretory granules. Paclitaxel
`
`also appears to act at the level of the genetic response to stimuli directing the cell to
`
`produce collagenase.
`
`The drug is also known to be effective in treating a number of other indications.
`
`I 0
`
`Paclitaxel is useful for treating surgical adhesions and post-surgical hyperplasias. In
`
`Alzheimer's disease treatment, paclitaxel has been used to stabilize microtubules
`
`destabilized by insufficient tau protein levels. U.S. Patent No. 5,580,898. Paclitaxel is also
`
`thought to be effective against polycystic kidney disease (PKD). Sommardahl et al. (1997)
`
`Pediatr. Nephrol. I I :728-33. Paclitaxel derivatives are also effective in treating psoriasis.
`
`I5
`
`EP 747385 and WO 96I3494.
`
`Other therapeutic agents have been successfully co-administered with paclitaxel.
`
`For example, Vitamin C can be used to increase the efficacy of paclitaxel. Kurbacher et al.
`
`(1996) Cancer Lett. 103: I83-I89. EP 78I552 and EP 7877I6 describe additional
`
`compounds that enhance paclitaxel activity. U.S. Patent No. 5,565,478 describes
`
`20
`
`combinational therapy of paclitaxel with signal transduction inhibitors for cancer treatment
`
`In treatment of autoimmune arthritis, paclitaxel has been administered with other
`
`antiarthritic drugs, such as an angiogenesis inhibitor. U.S. Patent No. 5,583,I53. Anilide
`
`derivatives have also been administered to sensitize multidrug-resistant cancer cells to
`
`paclitaxel. EP 6494IO. Paclitaxel can also be administered with antibodies specific to
`
`25
`
`cancerous cells. U.S. Patent No. 5,489,525. In breast cancer treatment, paclitaxel has been
`
`administered in combination with estramustine phosphate. Keren-Rosenberg et al. (1997)
`
`Sem. Oneal. 24 (Suppl. 3):S3-26-29. Paclitaxel and IGF-I (Insulin-like growth factor I)
`
`have been used together to treat peripheral neuropathy. U.S. Patent Nos. 5,648,335,
`
`5,569,648 and 5,633,228. Paclitaxel has also been successfully administered along with
`
`30
`
`doxorubicin, cyclophosphamide, and cisplatin. O'Shaughnessy et al. (1995) Breast Cancer
`
`Res. Treat. 33:27-37. P-glycoprotein blocker SDZ PSC 833, a cyclosporin derivative, has
`
`demonstrated a I 0-fold increase in oral bioavailability of paclitaxel in mice. Asperen et al.
`
`3
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`(1997) Brit. J Cancer 76:1181-1183. Essential oils have also been suggested to increase
`
`paclitaxel's bioavailability. U.S. Patent No. 5,716,928.
`
`The mechanism of paclitaxel action has been extensively studied and is summarized
`
`by Horwitz (1984) Pharm. Ther. 25:83-125. Paclitaxel can act by promoting tubulin
`
`5
`
`assembly into stable aggregated structures which resist depolymerization by dilution,
`
`calcium ion, cold, and several microtubule-disrupting drugs. Tubulin depolymerization is
`
`essential for cell division, and thus paclitaxel causes this process to cease. Schiff et al.
`
`(1979) Nature 277:665-667. Paclitaxel is unique in promoting tubulin polymer formation,
`
`whereas other anti-cancer drugs, such as vinblastine and colchicine, prevent this process.
`
`10
`
`As originally described in Wani et al. [(1971) J Amer. Chem. Soc. 93:2325-2327],
`
`paclitaxel can be purified via alcohol extraction from the Pacific yew tree, Taxus brevifolia.
`
`It is also present in other Taxus species, such as T baccata and T cuspidata. However,
`
`paclitaxel is found only in minute quantities in the bark of these slow-growing trees,
`
`causing concern that the limited paclitaxel supply will not meet the demand. Consequently,
`
`15
`
`chemists in recent years have attempted to find alternative or synthetic routes for producing
`
`paclitaxel. U.S. Patent No. 5,019,504 describes the purification of paclitaxel from tissues
`
`of T brevifolia grown in vitro. U.S. Patent No. 5,322,779 describes the production of
`
`paclitaxel from a fungus, Taxomyces andreanae, found in association with the yew tree.
`
`More recently, novel compounds have been suggested for use in enhancing plant
`
`20
`
`production of paclitaxel. U.S. Patent No. 5,710,099.
`
`Paclitaxel has also been synthesized from related compounds found in higher
`
`quantities in Taxus trees. These compounds include baccatin III, obtained from Taxus
`
`wood, and 10-deacetyl baccatin III, from Taxus leaves. Methods of preparing paclitaxel
`
`from these precursor compounds, which themselves lack anti tumor activity, have been
`
`25
`
`described. Greene et al. (1988) JACS 110:5917-5919; U.S. Patent Nos. 5,717,103,
`
`4,857,653, and 4,924,011 (Re. 34,277).
`
`Various synthetic routes and intermediates in paclitaxel synthesis have been
`
`described, including a route directed to the synthesis of the tricyclic taxane nucleus from
`
`commodity chemicals. Holton et al. (1994) J Am. Chem. Soc. 116:1597-1598, 1599-1600;
`
`30
`
`Nicolaou et al. (1994) Nature 367:630-634; and Danishefsky et al. (1996) J Am. Chem.
`
`Soc. 118:2843-59; and U.S. Patent Nos. 5,723,635 and 5,726,318. Additional compounds
`
`useful in paclitaxel synthesis have also been described. U.S. Patent No. 5,015,744
`
`4
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`describes the use of an oxazinone as a side-chain precursor for paclitaxel synthesis. U.S.
`
`Patent No. 4,876,399 describes an intermediate, 2,5-dihydroxy-2-patchoulene. U.S. Patent
`
`Nos. 5,523,219 and 5,705,671 describe additional intermediates.
`
`Paclitaxel itself has been chemically modified, sometimes producing compounds
`
`5
`
`with even greater antitumor activity than paclitaxel itself. U.S. Patent No. 4, 814,470.
`
`Cephalomannine, which differs from paclitaxel and baccatin III in the C-13 ester
`
`functionality, demonstrates activity against leukemia in animals. U.S. Patent No.
`
`4,206,221. Other paclitaxel derivatives include prodrug forms, in which paclitaxel is
`
`conjugated to cleavage spacer and sugar groups. EP 781778.
`
`10
`
`Some paclitaxel derivatives have been produced in attempts to address a significant
`
`problem limiting the utility of paclitaxel: paclitaxel is largely insoluble in water. This has
`
`created significant problems in developing suitable pharmaceutical formulations for human
`
`therapy both in terms of formulation and side effects. The problem is also a serious
`
`impediment for experimental research on paclitaxel and its clinical effectiveness.
`
`15
`
`Derivatives of paclitaxel, designed to have increased water solubility, include 2' - and/or 7-
`
`position paclitaxel esters, as described in U.S. Patent No. 4,960,790. Additional
`
`substitutions at the C-2' and C-7 positions were described by Magri et al. (1988) J Natural
`
`Products 51:298-306. 2' -succinyl paclitaxels are described in U.S. Patent No. 4,942,184;
`
`and sulfonated 2' -acryloyltaxol and sulfonated 2 '-0-acyl acid paclitaxel derivatives, in
`
`20
`
`U.S. Patent No. 5,059,699.
`
`Unfortunately, many of these more soluble derivatives reduce paclitaxel antitumor
`
`activity. A 2' -succinyltaxol, prepared by the treatment of paclitaxel with succinic
`
`anhydride, had decreased in vivo activity compared with paclitaxel, and a 2'-(t(cid:173)
`
`butyldimethylsilyl)taxol was essentially inactive. Magri et al. (1988). Other derivatives,
`
`25
`
`such as 2' -(~-alanyl)taxol, are unstable. Magri et al. (1988). Attempts to derivatize
`
`paclitaxel generally increase the molecule's size, which decreases its ability to passively
`
`diffuse through the cellular and nuclear membranes of cancerous cells.
`
`The insolubility of paclitaxel itself has yielded a further complication: it has elicited
`
`the widespread use of a toxic carrier. Paclitaxel is generally supplied through CTEP
`
`30
`
`(Cancer Therapy Evaluation Program), DCT (Division of Cancer Treatment), and NCI
`
`(National Cancer Institute, IND#2280) as a concentrated solution in 50% polyoxyethylated
`
`castor oil [Cremophor EL® (BASF)] and 50% dehydrated alcohol. This is then mixed with
`
`5
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`either a dextrose or sodium chloride solution prior to administration. Although Cremophor
`
`EL® is the industry-standard administration vehicle for paclitaxel, Cremophor EL® is
`
`itself toxic, causing idiosyncratic histamine release and anaphylactoid-like response.
`
`Cremophor EL® is also likely to be the cause of several side effects associated with
`
`5
`
`paclitaxel treatment, including cutaneous flushing, urticaria, dyspnea, bronchospasm, and
`
`hypotension. Runowicz et al. (1993) Cancer 71: 1591-1596; and Weiss et al. (1990) J
`
`Clin. Oneal. 8:1263-126. In studies with dogs, Cremophor EL® and its fatty acid
`
`constituents induced histamine release and hypotension within 10 minutes of
`
`administration. Lorenz et al. (1977) Agents Actions 7:63-67. Some tested animals died as a
`
`10
`
`result of this hypotension.
`
`Other organic carriers have been proposed for paclitaxel administration or used in in
`
`vitro paclitaxel preparations. Polyethylene glycol (PEG) has been suggested as a substitute
`
`emulsifier for paclitaxel, but PEG decreases the anti tumor activity of paclitaxel in murine
`
`tumor studies. Weiss et al. (1990) J Clin. Oneal. 8:1263-1268. Paclitaxel has also been
`
`15
`
`prepared in solution with dimethylsulfoxide [Kumar et al. (1993) Res. Comm. Chem. Path.
`
`Pharm. 80:337-344], which is itself toxic [Kamiya et al. (1967) Nippon Ganka Kiyo
`
`18:387-9; Sperling et al. (1979) Acta Ophthalmol. 57:891-8]. Polysorbate-80 was used in
`
`in vitro mixtures containing very low concentrations of docetaxel [Urien et al. (1996)
`
`Invest. New Drugs 14:147-151], but polysorbates are toxic, reducing locomotor activity,
`
`20
`
`inducing ataxia and hypotension, and increasing the activity of various carcinogens. Pesce
`
`et al. (1989) Ann. Clin. Lab. Sci. 19:70-3; (1984) J Am. Coll. Toxicol. 3/5: 1-82; and Varma
`
`et al. (1985) Arzneimittelforschung 35:804-8. Therefore, the sole use of these carriers to
`
`solubilize paclitaxel is not a desirable solution to the problem of developing therapeutically
`
`effective paclitaxel formulations.
`
`25
`
`In the absence of workable alternatives, and despite its toxicity, Cremophor EL®
`
`remains the standard vehicle used for paclitaxel administration to human patients.
`
`Documents demonstrating the universal use of Cremophor EL® in paclitaxel preparations
`
`and paclitaxel administration include: Einzig et al. (1991) Cancer Invest. 9:133-136;
`
`O'Shaughnessy et al. (1994) Breast Cancer Res. Treat. 33:27-37; Kawano et al. (1994) J
`
`30
`
`Toxicol. Sci. 19(suppl.1):113-122; Asperen et al. (1997) Brit. J Cancer 76:1181-1183;
`
`Sparreboom et al. ( 1998) Anti-Cancer Drugs 9: 1-1 7; Runowicz et al. ( 1993) Cancer
`
`71: 1591-1596; Sparreboom et al. (1998) Anal. Biochemistry 255: 171-175; Plasswilm et al.
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`6
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`(1998) Strahlentherapie und Onkologie 174:37-42; Xu et al. (1997) Hospital Pharmacy
`
`32: 1635-1638; Khan et al. (1997) Ann. Pharmacotherapy 31: 1471-1474; Michaud et al.
`
`(1997) Ann. Pharmacotherapy 31: 1402-1404; Zhang et al. (1997) Anti-Cancer Drugs
`
`8:696-701; Wilson et al. (1997)Ann. Pharmacotherapy 31:873-875; Reinecke et al. (1997)
`
`5
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`Eur. J Cancer Part A :United Kingdom 33:1122-1129; Kuangjing Shao et al. (1997) Anal.
`
`Chemistry 69:2008-2016; Bonfrer et al. (1997) Tumor Biology; Switzerland 18:232-240;
`
`Decorti et al. (1997) Cancer Chemother. Pharmacology 40:363-366; Ho et al. (1997)
`
`Neurosurgery 4016 :1260-1268; Terzis et al. (1997) British J Cancer 75:1744-1752;
`
`Kilbourn et al. (1997) Disease-a-Month 43 :282-348; Frasci et al. (1997) J Clinical
`
`10
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`Oncology 15:1409-1417; Sharma et al. (1997) International J Cancer 71:103-107;
`
`Georgiadis et al. (1997) Clinical Cancer Research 3 :449-454; Zhang et al. (1997) Cancer
`
`Chemother. Pharmacology40:81-86; EP 694303; WO 94/12031; and U.S. Patent Nos.
`
`5,733,888, 5,731,334, 5,719,265, 5,714,512, 5,703,117, 5,698,582, 5,696,153, 5,686,488,
`
`5,683,715, 5,681,846, 5,670,537, 5,665,761, 5,648,335, 5,648,090, 5,641,803, 5,633,228,
`
`15
`
`5,621,001, 5,616,608, 5,616,330, 5,614,549, 5,608,087, 5,604,202, 5,569,648, 5,583,153,
`
`5,580,899, 5,569,720, 5,565,478, 5,504,102, 5,496,846, 5,496,804, 5,478,860, 5,403,858.
`
`Numerous attempts have been made to produce aqueous solutions of hydrophobic
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`drugs. For instance, formulations of cisplatin combined with dextran, polyglutamic acid,
`
`DNA, proteins, hyaluronic acid, etc. were compared. It was found that many of these
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`20
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`excipients were unacceptable as they bound the drug too tightly and did not release it on
`
`administration or did not bind enough drug to produce a pharmaceutically acceptable
`
`formulation. DNA was in the category of excipients which bound too tightly. Proteins,
`
`including serum albumin, were found to bind limited amounts of drug, only a portion of
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`which was reversibly bound.
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`25
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`Albumins have been used as excipients as bulk stabilizers for a number of drug
`
`formulations, particularly biologicals such as interleukins and cytokines. Human serum
`
`albumin is a large component of interleukin-4 preparations. Meyer et al. (1994) Pharm.
`
`Res. 11: 1492-1495. Albumin has also been conjugated to drugs to increase uptake of the
`
`drug and derivatized albumins have been used to couple drugs and enhance uptake through
`
`30
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`the blood-brain barrier. Sinn et al. (1990) Nucl. Med. Biol. 17:819-827; Pardridge et al.
`
`(1990) J Pharmacol. Exp. Ther. 255:893-899; Flume et al. (1989) Pharm. Acta Helv.
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`64:351-352; and JP 61001622. WO 94/01090 describes broad formulations of hydrophilic
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`peptides and "sparingly water soluble" active compounds. Albumin is a cost-limiting
`
`component for use in drug stabilization. Thus, unless an unstable drug can be stabilized in
`
`some other fashion, albumin is not ideal as a bulk stabilizing agent. Further, native
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`albumin is being phased out of use as it may contain infectious agents such as prions.
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`5
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`Replacement with recombinant albumin may result in an even more costly product.
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`Therefore, in order to produce a commercially available, pharmaceutically acceptable
`
`albumin-bound drug, the drug must be bound reversibly to the albumin in a high molar
`
`ratio.
`
`The need remains for aqueous pharmaceutically acceptable formulations of
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`paclitaxel which are easy and inexpensive to prepare, produce fewer side effects, and in
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`which the drug retains high water solubility and activity.
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`SUMMARY AND OBJECTS OF THE INVENTION
`
`In one embodiment, the invention provides an optically clear, pharmaceutically
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`15
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`acceptable aqueous composition comprising paclitaxel or a derivative thereof, serum
`
`albumin or a fragment thereof, and a pharmaceutically acceptable vehicle. In various
`
`embodiments, the composition comprises no more than 10% organic solvent, and has a pH
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`of about 3.0 to about 4.8 (the pl of albumin). In various embodiments, the composition
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`comprises about 1 to about 10%, about 2 to about 8%, or about 4 to about 6% v/v
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`20
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`(volume/volume) organic solvent. In a preferred embodiment, the composition is essential
`
`free of organic solvent. The organic solvent is preferably an alcohol, most preferably
`
`ethanol. In various embodiments, the pH is about 3.0 to about 4.8, about 4.0 or less, about
`
`3.0 to about 4.0, or about 3.4 to about 3.8. In various embodiments, the ratio of paclitaxel
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`or derivative thereof to albumin is at least about 1 :5, at least about 1 :4, at least about 1 :2, at
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`25
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`least about 1: 1, or at least about 2: 1. In various embodiments, the serum albumin is
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`defatted, undefatted or a mixture of defatted and undefatted forms. In various
`
`embodiments, the serum albumin is mammalian, preferably human. In various
`
`embodiments, the serum albumin is at least about 50%, at least about 60%, at least about
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`70%, at least about 80%, or at least about 90% monomeric. In various embodiments, at
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`30
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`least about 70%, at least about 80%, at least about 85%, or at least about 90% of the
`
`paclitaxel or derivative thereof is bound to albumin. In another embodiment, the
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`composition is lyophilized. In another embodiment, the composition is reconstituted from a
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`lyophilized formulation. In various embodiments, the concentration of paclitaxel is greater
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`than about 25 µg/ml, greater than about 50 µg/ml, greater than about 100 µg/ml, greater
`
`than about 200 µg/ml, greater than about 300 µg/ml, greater than about 400 µg/ml, or
`
`greater than about 500 µg/ml. In another embodiment, the composition is coated onto an
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`5
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`implantable device such as a stent or wrap. In some embodiments, the device is catheter(cid:173)
`
`based and/or used in conjunction with surgery. In some embodiments, the coating prevents
`
`restenosis, local tumor growth or tissue over-growth and/or chronic inflammation.
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`The composition is characterized by having optical clarity for a length of time
`
`sufficient to administer to a patient or to process further (e.g., subject to drying). In another
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`embodiment, the composition is optionally dried and stored as a dried "storage-stable"
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`composition. The dried preparation of the composition is resolubilized prior to
`
`administration. In a preferred embodiment, the drying process is lyophilization. In one
`
`embodiment, the composition prior to drying comprises Mcllvaine buffer. In another
`
`embodiment, the lyophilized preparation of the composition is optionally reconstituted with
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`15
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`a physiologically acceptable vehicle, such as Mcllvaine buffer, water, a sugar solution such
`
`as dextrose or glucose, or certain saline solutions, including dilutions of saline. The
`
`reconstituted compositions can be essentially free of solvent, which can be removed in the
`
`lyophilization step. The resolubilized composition can be 2-10 times more concentrated
`
`than the original pharmaceutically acceptable composition, depending on the concentration
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`20
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`of paclitaxel in the pre-lyophilized composition. Thus, the invention encompasses a
`
`resolubilized composition which is optically clear for at least 8 hours after reconstitution.
`
`The composition comprises less than 10% organic solvent and has a pH of about 3 .0 to
`
`about 4.8 upon reconstitution, at least about 70% of the paclitaxel introduced into the
`
`composition is bound to the serum albumin, and the paclitaxel concentration in the
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`25
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`composition is at least 50 µg/ml. The invention further encompasses methods of
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`administration of the reconstituted composition wherein a therapeutically effective amount
`
`of paclitaxel can be administered as a 1 to 3 hour (or greater) injection or as a bolus.
`
`In another embodiment, the invention encompasses a method of treatment,
`
`comprising administering to a patient a therapeutically effective amount of an optically
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`30
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`clear, pharmaceutically acceptable aqueous composition comprising paclitaxel or a
`
`derivative thereof, serum albumin and a pharmaceutically acceptable vehicle, as described
`
`above. The indication to be treated with the composition can include any indication known
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`in the art to be treatable with paclitaxel, including, but not limited to, cancer. Preferably,
`
`the cancer affects cells of the bladder, blood, bone, brain, breast, cervix, colon, epithelium,
`
`digestive tract, head/neck, kidneys, liver, lung, mouth, ovaries, pancreas, prostate gland,
`
`skin, stomach, testicles, or tongue. The indication can also include, but is not limited to,
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`paclitaxel-treatable indications such as Alzheimer's disease, kidney disease, peripheral
`
`neuropathy, psoriasis, restenosis, rheumatoid arthritis, systemic lupus erythematosus,
`
`surgical adhesions, or tissue overgrowth after surgery. Preferably, the patient is a mammal.
`
`More preferably, the mammal is a human.
`
`The composition and methods of use thereof can optionally further comprise an
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`10
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`additional biologically active ingredient, including but not limited to those known to
`
`function synergistically with paclitaxel. In various embodiments, the additional agent
`
`includes, but is not limited to, G-CSF (granulocyte colony-stimulating factor), GM-CSF
`
`(granulocyte macrophage colony-stimulating factor), IL-4 (interleukin 4), IGF-I, analide
`
`derivatives, antiarthritics (e.g., an angiogenesis inhibitor), antibodies specific to cancer
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`15
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`cells, antineoplastics (e.g., carboplatin, cyclophosphamide, estramustine phosphate, and
`
`etoposide ), doxorubicin, immunosuppressants (e.g., cisplatin and cyclophosphamide ),
`
`steroidal and non-steroidal hormone (e.g., cortisone), transduction inhibitors, and vitamins
`
`(e.g., vitamin C). The composition can further comprise low concentrations of excipients
`
`such as polyethylene glycol, detergents, organic solvents, or organic or inorganic acids.
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`20
`
`In another embodiment, the invention encompasses a method of making an optically
`
`clear, pharmaceutically acceptable aqueous composition comprising paclitaxel or a
`
`derivative thereof, serum albumin and a pharmaceutically acceptable vehicle, as described
`
`above, comprising the steps of preparing a solution of the paclitaxel or a derivative thereof,
`
`preparing a solution of serum albumin, and slowly combining the solutions. Due to stable
`
`25
`
`binding of Ptx to serum albumin, the rate of addition of the Ptx solution to the albumin
`
`solution can be decreased to assist in more optimal loading of Ptx onto albumin. The
`
`paclitaxel solution can, for example, be added dropwise at a controlled rate; this rate can
`
`be, for example, at about 0.1 to 10 ml/min, e.g., 1 ml/min or slower, and the drop size can
`
`be 8 to 20 µ1. In various embodiments, the ratio of paclitaxel or derivative thereof to
`
`30
`
`albumin is at least about 1: 1 or at least about 2: 1, and the solutions are combined at a
`
`temperature below room temperature, about 2°C to 8°C, or about 4°C. In various
`
`embodiments, the ratio of paclitaxel or derivative thereof to albumin is at least about 1: 5, at
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`least about 1 :4, at least about 1 :2, at least about 1: 1, or at least about 2: 1. It is anticipated
`
`that ratios of 3: 1 and possibly even 4: 1 can be achieved according to the invention
`
`described herein, by controlling the rate of addition of the paclitaxel to the albumin solution
`
`to a degree that does not interfere with continued stability during processing.
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`5
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`Preferably the paclitaxel is "optimally concentrated." This term means that the
`
`paclitaxel concentration in the composition allows a solvent concentration of 1 - 10% v/v.
`
`The molar ratio of paclitaxel:albumin and the final concentration of paclitaxel in the
`
`albumin solution are optimized, such that the paclitaxel remains in solution for a length of
`
`time practical for administration or lyophilization/reconstitution. We have found that the
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`10
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`highest concentrations of paclitaxel and optimal molar ratios are achieved with final
`
`ethanol concentrations in the 1-10% range, more preferably in the 2-8% rang