CA2202531C - Lyophilizate of lipid complex of water insoluble camptothecins - Google Patents
Lyophilizate of lipid complex of water insoluble camptothecins Download PDFInfo
- Publication number
- CA2202531C CA2202531C CA002202531A CA2202531A CA2202531C CA 2202531 C CA2202531 C CA 2202531C CA 002202531 A CA002202531 A CA 002202531A CA 2202531 A CA2202531 A CA 2202531A CA 2202531 C CA2202531 C CA 2202531C
- Authority
- CA
- Canada
- Prior art keywords
- composition
- camptothecin
- phospholipid
- glycerol
- water insoluble
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
- A61K9/1277—Preparation processes; Proliposomes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/13—Amines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biophysics (AREA)
- Organic Chemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Dispersion Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dermatology (AREA)
- Hematology (AREA)
- Oncology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
The present invention is directed to a pharmaceutical composition comprising a lyophilizate of a non-liposomal complex of camptothecin and phosphilipid for the treatment of cancer.
Description
LYOPHILIZATE OF LIPID COMPLEX OF WATER
INSOLUBLE CAMPTOTHECINS
Background of the Invention The present invention relates to a pharmaceutically acceptable dosage form for water insoluble camptothecins, for example 9-aminocamptothecin, and, more particularly 9-amino 20(S) - camptothecin (hereafter 9-AC), 9-amino-20-(R,S)-camptothecin and 9-amino-20 (R) -camptothecin which can be reconstituted and administered to a patient intravenously or subcutaneously or formulated for oral use in the treatment of cancer and other diseases.
Camptothecin and analogues thereof, display antitumor activities . against colon cancer, leukemia, and experimentally transplanted carcinoma such as leukemia L-1210 in mice or Walker 256 tumors in rats (See Potsmiesel, M. et al "Preclinical Studies of DNA Topoisomerase I-targeted 9-amino and 10, 11 Methylenedioxy Camptothecins," in DNA Topoisomerases in Cancer, edited by Milan Potmesil and Kurt Kohn, Oxford Univ. Press, New York, NY, p. 299-311 (1991)). Camptothecin is a pentacyclic alkaloid including a characteristic fused 5-ring system of quinoline (rings A and B), pyroline (ring C), a-pyridone (ring D) and a six-membered lactone (ring E). The intact lactone ring, ring E, and hydroxyl group at , position 20 have been found to be essential to its antitumor activity.
Studies of camptothecin analogs have suggested a correlation between the compound's ability to induce DNA
breakage and its antitumor ac~ivity. It has a unique mechanism of action whit'.~. produces DNA damage.in the presence of topoisomerase I, a monomeric enzyme that is capable of altering DNA topology in euicaryotic cells.
Topoisome.rase T_ binds to the JNA to allow tine double helix to unwind and subsequently reseals the break before WO 96111669 . PGT/ITS95I14105 ' dissociating from the DNA strand.. Camptothecin is believed to.bind to and stabilize a rovalent DNA-topoisomerase I
l .. helix is broken and complex in which one strand of the DNA
thereby prevent the DNA from recombining.
.. The therapeutic use of camptothecin and its :
:~i . .~
.;.;; analogs has~been severely. limited by their poor water solubility and high toxicity. A number cf attempts have been made to reduce the. toxicity of camptothecin without reducing its antitumor activity through the development of w ; derivatives. Camptothecin derivatives substituted,at the 5-, 7-, 9-, 10-, and 11-positions have been '_nvestigated. At I least three camptothecin derivatives ar'e .n various stages w . , of clinical development, namely: 7-ethyl-[14-(1-piperidirio)-?-piperidine] carbonyloxycamptothecin (CPT-11);
20-(S)-camptothecin; 10-[(dimethylamino) methyl]-4-et:hyl--~ 4,9-dihydroxy-1H=pyrano[3',4':6,7]indolizino[1,2-.:~
b] quirioline-3 ,14 (4H,12H) dione monohydrochloric~e (topotecan .! hydrochloride); and 9-amino-20(s). - camptothecin.
9-AC is quite water insoluble..!0.002 mg/ml). This makes it difficult to formulate as a sterile, storage 1~: .
.:..y.:~_ stable, dosage form. In studies carried 'out by the ~" , National Cancer Institute (NCI) (Dahust et al., Journal of Clinical Oncology, 14, 1236-1244(1996)) . 9-AC was formulated in an organic solvent, dimethylacetaanide (DMA), to overcome the solubility Limitations of the drug. The NCI formulation consists of 5 mg of 9-AC in 1 ml of DMA. At the point of use, the product is diluted using a diluent consisting of polyethylene glycol 400, USP, and phosphoric acid. The . , ~ diluent is added to the NCI formulation in an amount of 49 ml diluent to 1 ml formulation.
The NCI Formulation has drawbacks which make the - ' formulation izconveaient for commercial Lse. Because the DMA attacks.._rubber stoppers, the produc~~~cannc~ be supplied ii-n stoppered vials .and must be supplied -:~ ainpuies . Ampuies are inconvenient to use because they must be scored and broken to open them and this presents some risk of injury and contamination to workers. Additionally, glass chips in the product from breaking open the ampuies need to be filtered out.
DMA is not a desirable vehicle for intravenous (IV) administration and a potential source of toxic side effects. DMA has an LDso value of 5.4 ml/kg, which is a factor which also must be considered in administering the drug.
Another approach to designing camptothecin dosage forms which has been investigated involves the use of liposomes. T. G. Burke et al. in Biochemistrv 1993, 32, 5352-64 (1993) suggest using liposomes as a drug delivery system for camptothecin. Burke et al. found that camptothecin binds with dimyristoylphosphatidylglycerol (DMPG) lipids and dimyristoylphosphatidylcholine (DMPC) lipids and is stable in both DMPC and DMPG liposome bilayers. They postulate that the lactone ring penetrates the liposom~ layer. The liposome-associated camptothecin showed stabilization of the lactone ring.
No satisfactory pharmaceutically acceptable formulation cf 9-AC is currently available for administration to humans. There is a need for a stable pharmaceutical dosage form which may be conveniently administered to a cancer patient while retainincr the structural elements that are essential for 9-AC's pharmacological activity.
Summary It is an object of the present invention to provide a pi:armace~~tically acceptable dosage form of 9-AC or anotY~er water insoluble camptothecin. It is another object of the present invention to provide lipid complexes of 9-AC ' and other water insoluble camptothecins. It is still another object of the invention to provide lyophilizates of three lipid complexes. The lyophilizate can be reconstituted with water, saline, or another electrolyte to give a colloidal dispersion for intravenous or subcutaneous administration or can be formulated into a paste or filled into a soft gelatin or hard gelatin capsule for oral administration. Previously, camptothecins have not been administered subcutaneously because they are necrotic.
However, it appears that the lipid complex may sufficiently slow the release of water insoluble camptothecins that subcutaneous administration is possible.
While the invention will hereafter be described with respect to the preparation of lipid complexes and lyophiiizates of lipid complexes of 9-AC, those skilled in the art will appreciate that the methods taught herein are also applicable to the preparation of lipid complexes and lyophilizates of other camptothecins which are considered water insoluble such as cam~tothecin itself.
In accordance with the present invention, a lyophilizate of a phospholipid complex of 9-AC is prepared which can be reconstituted with pharmaceutically acceptable aqueous diluent such as water for injection and which, in comparison to solutions of 9-AC in dimethylacetamide, is less toxic, more stable and particularly importantly, its formulation and administration are not limited by the , solubility of 9-AC.
In accordance with the invention, the lyophiiizate , is prepared by a process comprising the steps of preparing a solution of 9-AC in a first organic solvent, preparing a solution of a phospholipid ~__ a second organic solvent, mixing the phospholipid solution and the 9-AC solution, adding water to the mi~sced solutions to cause formation cf a lipid complex of 9-AC, removing the first and second orcranic solvents to provide a dispersion of the lipid complex ~n water as an aqueous phase, dissolving a pharmaceutically acceptable excipient in the aqueous phase of the dispersion, and lyophilizing the dispersion of the lipid complex to form a lyophilizate. This lyophilizate forms a colloidal dispersion when reconstituted with a physiologically acceptable aqueous diluent.
In accordance with a preferred embodiment ef the present invention, the lyophilizate is prepared by a method comprising the steps of forming a concentrated solution of 9-AC in dimethyl sulfoxide, forming a concentrated solution of the phospholipids DMPC and DMPG in chloroform, mixing the phospholipid solution and the 9-AC solution, adding an aqueous solution such as water for injection to form the lipid complex of 9-AC and provide a dispersion of the 'ipid complex in water as an aqueous phase, sparging and diafiltering the dispersion to remove the solvents, red~;cing the particle size of the dispersion of the lipid complex, adding an aqueous solution of mannitol as a pharmaceutically acceptabl' lyophilization excipient to the dispersion, and lyophilizing, wherein a iyophilizate is obtained which :aon reconstit:a ing with water provides a colloidal dispersicn of a 9-AC lipid complex.
In accordance with a further embodiment cf the present invention, a lyophilized composition containing :-aminecamptothecin or another water insoluble camptcthecin is provided hhich comprises 9-AC, a phospholipid and a pharmaceutically acceptable lyophilization excipient.
Detailed Description of the Invention The term "water insoluble camptothecin" as used herein means camptothecin or another pentacyclic alkaloid having the aforementioned characteristic fused 5-ring system of camptothecin having a solubility in water which is less than .O1 mg/ml at 23°C.
The term "lipid complex" is an art recognized term. Lipid complexes are characterized by a noncovalent bond between the lipid and the camptothecin which is observed as a phase change in differential scanning calorimetry.
The term "pharmaceutically acceptable aqueous diluent" as used herein refers to water for injection, saline, and other known aqueous vehicles.
The term "lyophilization excipient" refers to a substance which is added to a solution prior to lyophilization to enhance characteristics such as the color, texture, strength, and volume cf the cake. Sxamples of lyophilization excipients are provided below 9-AC has been prepared by reduction of 9-nitrocamptothecin with a reducing agent such as a combination of tin or iron with a mineral acid. (See U.S.
Patent 4,604,463 to Miyasaka et al.) The preparation of 9-amino-20(S)- camptothecin and ~-amino-20(R,S)-camptothecin is described in U.S. Patents 5,106,742 and ,225,404. Other water insoluble camptothecins are known it the art and their preparation is described in the literature. , 9-Amino-20(S)-camptothecin has a chemical formula of C2o H,., N3 OQ and the structure:
o' A B CAN /
11 ~13 ~ 2 3 t5 T_n accordance with the invention a concentrated solution ef 9-AC in an organic solvent .s prepared. The most typical example of the solvent used to prepare this solution is dimethyl sulfoxide (DMSG). However, other organic solvents such as dimethyl formamide can be used. Useful solvents must form stable solutions wit:: the camptothecin analogue, e.g., the solvent must not interact with, destabilize, and/or deactivate the drug. In addition, the solubility cf the camptothecin analogue in the solvent must also be high enough that the camptothecin analogue can be dissolved ~r. amounts sufficiently high to form commercially useful quantities cf the lipid complex and the solvent should be capable of being removed easily from an aqueous dispersion c~ the lipid comr~iex as described hereinafter.
Preferably, a solution having a concentration of about 5 to SO mg/ml, preferably about 20 to 40 mg/rl and most preferably ~0 mg/ml camptothecin is used. The concentratior.
may vary depending upon the nature of the solvent and temperature, but it is important to use a concentrated solution o~ the camptothecin ir. preparing the liquid/camptothecin complex. "'his mini;tizes the amount of solvent that must be removed ~aLer In t-:e prC a c ss, anti also assists in forcing the camptotheci: out oT solution and into liQUid/camDtothecin complex formation with the ~ipid with tine addition c~ water.
The organic solvent used to prepare tha solution of the phospholipids should meet similar requirements to those outlined for the camptothecin solvent. It must be compatible with the phospholipids and not destabilize them or the camptothecins. In addition, the lipids should be soluble enough in the solvent so as to be able to introduce enough of the lipid to form the complex yet minimize the amount of solvent that must be removed later. A solvent which can be readily removed from the dispersion of the lipid complex is most preferred. The solvent most typically used to prepare this solution is chloroform or methylene chloride. Typically the concentration of this phospholipid solution will range from about 10 to 250 mg/ml.
Phospholipids are amphipathic in nature, i.e., the molecules have a hydrophobic tail such as a long chain hydrocarbon, and a hydrophilic head. In an aqueous medium, such as water or saline, the tails align with each other, away From the aqueous molecules, while the heads face outward into the aqueous phase. It is this nature of the phOSphOlipids that makes them very useful for formulating highly insoluble drugs like 9-AC.
The phospholipids used in tine invention are selected such that their phase transition temperature is about equal to or below the body temperature or 37~C and the complex releases the drug in the body. Representative examples of useful phospholipids include synthetic phospholipids DMPC, DMPG, dipalmitoylphosphatidylcholine (DPPC;, dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylcholine (DSPC), or distearoylphos-phatidylglycerol (DSPG), or a combination thereof. Other examples of phospholipids can be found in the CRC Handbook of Lipid Bilavers by Marsh, M.A., CRC Press (1990). When _g_ W O 96!11669 PCTIUS95I14105 s DMPC and DMPG are used in a ratio of DMPC to DMPG of about ' 7:3 they mimic the cell membrane.
The lipid solution is added to the 9-AC solution such teat the weight ratio of the 9-AC to lipid is about 1:80 to 1:5, preferably about 1:80 to 1:10, more preferably, about 1:60 to 1:10 and still more preferably about 1:45 to 1:25.
In some applications, it has been found desirable to add cholesterol or its hemisuccinate derivatives to the lipid complex. The cholesterol is believed to cause the biiayers to pack more closely and thereby slow the release of the drug. This approach may be particularly desirable with subcutaneous formulations where severe necrosis can result if the drug is delivered too auickly. The cholesterol is added to the phospholipid solution. The cholesterol may be used in an amount of about 0.5 to 15 parts per 100 parts phospholipid.
Once the solvent solutions of the lipids and camptcthecin have been mixed, water or an aqueous solution is added rapidly and with stirring for several minutes tc the mixture. Addition cf the water is believed to cause the 9-AC and the lipid to come out cf their respective solvent sol~:tions and complex with each other. The Ovate- .s preferably added in an amount such that the 9-AC is prese.~.t in an amount of about .05mg to 0.5mg per 100m1 water. It is desirable to limit the amount of water to minimize the amount of water which must be removed during the lyophilization process. Higher amounts of water are undesirable because they increase the amount of water that must be removed d~,~ring the subseauent lyophiiization process. It is believed complexat~c_~_ may be complete in about 3C minutes. However, it is desirable to stir the _g_ WO 96/11669 PCT/US95/141~5 dispersion for about one hour to insure complexation is complete.
The lipid complex dispersion described above, is treated to remove the solvents. Any of a variety of techniques can be used for this purpose. For example, it has been found that the chloroform can be removed if the dispersion is sparged with a gas such as nitrogen. A
diafiltration process (also known as a tangential flow filtration process) is used to remove the DMSO. A cartridge of hollow fiber tubes having a pore size of 5 to 150 kiiodaltons can be used. A diafiltration cartridge is available from A/G Technology Corporation cf Needham, Massachusetts under the tradename XPRESS, which can be used.
Other techniques which can be used include centrifugation.
A pharmaceutically acceptable iyophilization excipient is dissolved in the aqueous phase of the dispersion. Mannitol is typically used as the excipient but other excipients which do not interact with the drug or the lipid complex may be used. Sodium or potassium phosphate, citric acid, tartaric acid, gelatin, and carbohydrates such as mannitol, lactose, dextrose, dextran, hetastarch, etc.
are common examples of excipients which are also believed ~..
be useful herein. The excipients can be used alone or in combination to provide a cake of good quality which readily disperses in water upon reconstitution.
The excipients are typically added to the dispersion as solutions in water. Again, it is desirable to use concentrated solutions to minimize the amount of water .
for removal by lyophilization. The amount of the excipient is adjusted in a manner that is well known in the art to provide a cake which does not crack or shrink and is porous so that it readily dissolves and has a good appearance.
Mannitol has been found to be useful. Mannitoi is added tc WO 96/11669 ~ PCT/US95/14105 j the dispersion as a solution having a concentration of aboit to 150 g/ml. Mannitol is added in ar_ amount of about 1 to j 100 parts by weight per 1 part 9-AC.
After removing the solvents and adding the excipient, the dispersion is passed through a homogenizes (e.g.,. a Tekmar rotor/stator homogenizes, Model T25, or a microfluidics submerged jet homogenizes, Model M110Y). As a general rule, the smaller the particle size of the dispersion, the faster the formulation can be dried during 'i the lyophilization cycle. A dispersion having a particle size distribution ranging from about 10 to 500nm and v averaging about 250nm has been found to be satisfactory for j lyophilization. The optimum particle size may vary depending-on the mode of administration.
A typical lyophilization cycle useful in accordance with the present invention is provided below.
i ~ The cycle may be varied depending upon the equipment and facilities available in a manner well known in the art.
The homogenized formulation can be poured into vials of a 5 to 50m1 nominal volume. ':he vials are placed .. into a lyophilization chamber at about = °C. The vial size will usually be selected such that eacvial contains a single dosage cf the 9-AC: The temperature of the chambe r is reduced to -30°C over a period of one hour after which the temperature is maintained at -30°C for about four hours.
The pressure in the iyophilization.chamber is then reduced to 200-250 microns of pressure for the remainder of the cycle.. After reducing the pressure in the chamber, the temperature is ramped up to +25°C over a period of fifteen hours and the product is held at 25°C _:,= five hours. The temperature then is ramped up to +40°C over a period of 20 minutes and held at 40°C for two hours. The lyophilized * trademark -11-product preferably has a final moisture content of less than about 5o and typically about 1 to 2%. ' For intravenous or subcutaneous administration, the lyophilizate can be reconstituted using aqueous vehicles such as water, saline or another electrolyte. The lyophilized product with the addition of water provides a colloidal dispersion of the lipid complex in an aqueous solution of the excipient. Neither the 9-AC nor the lipids are soluble in water. A colloidal dispersion consists of at least two discrete phases. The first is a dispersed or internal phase. The second is a continuous or external phase. Systems in the colloidal state contain one or more substances that have at least one dimension in the range of 10-100A to a few microns. See pp. 272-4 in Chapter 19, Disperse Systems, Reminaton's Pharmaceutical Sciences, 18th Edition, 1990, Mack Publishing Company, Easton, PA 18042.
In the colloidal dispersions of the present invention, the dispersed cr internal phase comprises particles of 9-AC
lipid complex having a particle size in the range of lOnm to 1000nm. In selecting the aqueous vehicle, it is recommended to use one having a specific gravity about equal to the lipid complex (est. 1.09gjcc; to minimize the tendency for the dispersion to separate. The lyophilizate of the lipid complex can be reconstituted with water, saline, or another pharmaceutically acceptable aqueous diluent for intravenous administration. Upon reconstitution a dispersion is obtained which is suitable for injection. The lyophilizate can also be administered orally as an aqueous dispersion or , as a paste. While camptothecins are not generally administered subcutaneousl,r because they cause necrosis, it has been observed that the lipid slows the release of the camptothecin into the tissue making it potentially feasible to administer the lipid complex subcutaneousiy.
WO 96f11669 PCTlLIS9S/I4I05 For oral administration, the lyophilizate can be reconstituted tJ form an oral dispersion or formu~.ated into a paste. Alternatively, the lyophilizate can be filled into a soft gelatin capsule for oral administration.
Suitable dosages for 9-AC and camptothecins range from about 35 to 250 mg/m~/hour. The drug is preferably administered as a continuous infusion over 3 to 21 days using programmable continuous infusion ambulatory pump. It is anticipated that the drug will be administered with granulocyte colony stimulating factor (GCSF).
While it is contemplated herein that the lipid/camptothecin complex will be lyophilized to enhance its stability, it will be appreciated that the lipid/camptothecin complex is pharmaceutically active and can be formulated into a dosage form for oral, intravenous or subcutaneous administration without lyophilization.
Formulation aids such as antibacterials and antioxidants can be used to enhance the stability of the complex.
The invention will now be described in more detail with reference to the following non-limiting examples.
Examples ~-8 Lyophilizates of 9-AC were prepared using solvents, phospholipids and excipients in the amounts shown in Table 1. In each case, solutions were prepared by dissolving 9-AC in the amounts shown in DMSO, and, dissolving DMPC and DMPG in chloroform. The solutions were mixed and water for injection was added. The resulting dispersions were stirred for about one hour and cloroform was removed by sparging with nitrogen for about 95 min.
DMSO was removed by centrifuging the dispersion, removing the supernatant, and re-suspendir~g the "plug" in water for .E:':.:.
injection. tIn some examples, Millipore or AG Techno~ogy tangential flow filters were used to remove DMSO.) An aqueous solution of the excipient was then.
added to the dispersion and the dispersion was homogenized *.
>,;~ using a ULTRA TURREX homogenizes operating at approximately.
10,000 rpm, and room temperature. The homogenates were then lyophilized using the protocol described above or a similar process. , .
i --i Facamnie 1 2 3 4 ) 6 7 8 Na.
9 10.4 0.4 0.4 0.4 0.4 0.4 0.2 0.2 AC ma me mg mo mg mg mg ma -DMSO 0.01 0.01 0.01 8.7 '8.7 to 5 ml 5 ml ml ml ml ml ml ml - DMPC 126 24.2 3.0 22.4 22.4 22.4 11.2 8 ma~
~ ma ma mg ma m ma ma~
:-.,.-. DMPG _ 10.4 1.25 . 9.6 9.6 4.9.mg8 ma 11:3, ma mg 9.6 mg ma.
mg ma - Chloroform0.25 0_25 0.31 0.2 0.2 0.2 O.l 0.1 ml ml ml :~1 ml ml mg ml Mannitol 15.1 14.55 14 ma 15 15 20 120 20 ma ma ma ma ma ma ma ~ , Sodium 8.9 V8.9 mg' mg Citrate .. Cil_ = ~ 14.7 1y.7 .- ACid ma ~a WFI -Lyochilizate~0.4 10.4 0.4 0.2 ~0.2 12 1 ma/
mg/ mg/ mg/ mgl mg/ mg/
vial vial vial vial , vial vial vial The. lydphilizate obtained in Example 1 was placed on - stability testing at -4°C, 27°C, and 37°C. The initial assay showed~0.299 mg 9-AC-per,vial. the.results of the study to date are shown in Table 2.
* trademark WO 96/11669 PCTliIS95/I4I05 r Percent Initial Assav 4'C 27'C 37'C
1 week ~ 99.3 97.7 99.7 2 weeks ~ 100.3 103.3 104.3 1 month ~ 103.7 97.7 102.3 2 months ~ 101 102.3 102.3 months ~ 106.4 105.7 104.7 7 months I 106.4 104.2 ---The results of the study show no evidence of deterioration in the lyophilizate of the lipid/camptcthecin complex.
Example 9 Studies in animals have shown that the lyophilizates of 9-AC of the present invention exhibit antitumor activity in vivo. Colloidal dispersions of the lyophilizate were prepared by dispersing 1 mg cf the lyophilizate cf Example 6 above in 10 mi of water for injection. The dispersions were evaluated against the HT29 human colon tumor xenograft in female athymic mice. Both intravenous and oral administrations were used. Tumor xenografts were allowed to grow to abou~ 200 mg before initiation of chemotherapy. The size c. the tumor was determined based on tumor volume. Treatment comparisons were based on time (days) to three tumor doublings (TTTD).
The results are summarized in the Table 3. A high level of citrate was present in the formulation curing the first course of treatment and resulted in a nigh incidence of vehicle-related mortality at the highes~ I.v. dose levels.
The formulation was orally active on a Q2Dxl4 schedule with no toxicity.
I.V. and P.O. Activity of 9-AC vs. HT29 Xeno r CDF
Schedule mg/kg Dose TTTD Deaths 0 26.8 0 I.V. Bolus 1.8 34.2 0 Q4Dx6 2.7 38.9 2a (6 doses) ~ 4.0 ~ 25.3 ~ 6a I.V. Bolus 0 26.3 0 Qdx4 0.9 35.4 0 Days 12,19,26 1.33 46.4 2b 12 doses 2.0 50.9 7b 0 27.0 0 P.~. 0.67 35.6 0 1.0 33.7 0 Q2Dx14 1.5 42.8 0 CDF = Collidal dispersion formulation (lipid formulation); TTTD = Time (days) to 2 X doubling of tumor size Start Treatment Day 12 for I.V., Day 13 for P.O.; N =
10/group First dose or CDF contained citrate buffer. Most deaths attributed to vehicle. Second and remaining doses were citrate-free First course of DMSO/intralipid containing citrate buffer. Most deaths attributed to vehicle. Second and third courses were citrate-free 1 partial regression SUBSTITUTE SHEET (RULE 2G) i Having described the invention in detail and by reference to preferred embodiments thereof, it will be apparent that modifications and variations are possible ~;- ~ without departing from the scope of the invention defined in the appended claims.
. -16/i-SUBSTITUTE SHEET (RULE 26)
INSOLUBLE CAMPTOTHECINS
Background of the Invention The present invention relates to a pharmaceutically acceptable dosage form for water insoluble camptothecins, for example 9-aminocamptothecin, and, more particularly 9-amino 20(S) - camptothecin (hereafter 9-AC), 9-amino-20-(R,S)-camptothecin and 9-amino-20 (R) -camptothecin which can be reconstituted and administered to a patient intravenously or subcutaneously or formulated for oral use in the treatment of cancer and other diseases.
Camptothecin and analogues thereof, display antitumor activities . against colon cancer, leukemia, and experimentally transplanted carcinoma such as leukemia L-1210 in mice or Walker 256 tumors in rats (See Potsmiesel, M. et al "Preclinical Studies of DNA Topoisomerase I-targeted 9-amino and 10, 11 Methylenedioxy Camptothecins," in DNA Topoisomerases in Cancer, edited by Milan Potmesil and Kurt Kohn, Oxford Univ. Press, New York, NY, p. 299-311 (1991)). Camptothecin is a pentacyclic alkaloid including a characteristic fused 5-ring system of quinoline (rings A and B), pyroline (ring C), a-pyridone (ring D) and a six-membered lactone (ring E). The intact lactone ring, ring E, and hydroxyl group at , position 20 have been found to be essential to its antitumor activity.
Studies of camptothecin analogs have suggested a correlation between the compound's ability to induce DNA
breakage and its antitumor ac~ivity. It has a unique mechanism of action whit'.~. produces DNA damage.in the presence of topoisomerase I, a monomeric enzyme that is capable of altering DNA topology in euicaryotic cells.
Topoisome.rase T_ binds to the JNA to allow tine double helix to unwind and subsequently reseals the break before WO 96111669 . PGT/ITS95I14105 ' dissociating from the DNA strand.. Camptothecin is believed to.bind to and stabilize a rovalent DNA-topoisomerase I
l .. helix is broken and complex in which one strand of the DNA
thereby prevent the DNA from recombining.
.. The therapeutic use of camptothecin and its :
:~i . .~
.;.;; analogs has~been severely. limited by their poor water solubility and high toxicity. A number cf attempts have been made to reduce the. toxicity of camptothecin without reducing its antitumor activity through the development of w ; derivatives. Camptothecin derivatives substituted,at the 5-, 7-, 9-, 10-, and 11-positions have been '_nvestigated. At I least three camptothecin derivatives ar'e .n various stages w . , of clinical development, namely: 7-ethyl-[14-(1-piperidirio)-?-piperidine] carbonyloxycamptothecin (CPT-11);
20-(S)-camptothecin; 10-[(dimethylamino) methyl]-4-et:hyl--~ 4,9-dihydroxy-1H=pyrano[3',4':6,7]indolizino[1,2-.:~
b] quirioline-3 ,14 (4H,12H) dione monohydrochloric~e (topotecan .! hydrochloride); and 9-amino-20(s). - camptothecin.
9-AC is quite water insoluble..!0.002 mg/ml). This makes it difficult to formulate as a sterile, storage 1~: .
.:..y.:~_ stable, dosage form. In studies carried 'out by the ~" , National Cancer Institute (NCI) (Dahust et al., Journal of Clinical Oncology, 14, 1236-1244(1996)) . 9-AC was formulated in an organic solvent, dimethylacetaanide (DMA), to overcome the solubility Limitations of the drug. The NCI formulation consists of 5 mg of 9-AC in 1 ml of DMA. At the point of use, the product is diluted using a diluent consisting of polyethylene glycol 400, USP, and phosphoric acid. The . , ~ diluent is added to the NCI formulation in an amount of 49 ml diluent to 1 ml formulation.
The NCI Formulation has drawbacks which make the - ' formulation izconveaient for commercial Lse. Because the DMA attacks.._rubber stoppers, the produc~~~cannc~ be supplied ii-n stoppered vials .and must be supplied -:~ ainpuies . Ampuies are inconvenient to use because they must be scored and broken to open them and this presents some risk of injury and contamination to workers. Additionally, glass chips in the product from breaking open the ampuies need to be filtered out.
DMA is not a desirable vehicle for intravenous (IV) administration and a potential source of toxic side effects. DMA has an LDso value of 5.4 ml/kg, which is a factor which also must be considered in administering the drug.
Another approach to designing camptothecin dosage forms which has been investigated involves the use of liposomes. T. G. Burke et al. in Biochemistrv 1993, 32, 5352-64 (1993) suggest using liposomes as a drug delivery system for camptothecin. Burke et al. found that camptothecin binds with dimyristoylphosphatidylglycerol (DMPG) lipids and dimyristoylphosphatidylcholine (DMPC) lipids and is stable in both DMPC and DMPG liposome bilayers. They postulate that the lactone ring penetrates the liposom~ layer. The liposome-associated camptothecin showed stabilization of the lactone ring.
No satisfactory pharmaceutically acceptable formulation cf 9-AC is currently available for administration to humans. There is a need for a stable pharmaceutical dosage form which may be conveniently administered to a cancer patient while retainincr the structural elements that are essential for 9-AC's pharmacological activity.
Summary It is an object of the present invention to provide a pi:armace~~tically acceptable dosage form of 9-AC or anotY~er water insoluble camptothecin. It is another object of the present invention to provide lipid complexes of 9-AC ' and other water insoluble camptothecins. It is still another object of the invention to provide lyophilizates of three lipid complexes. The lyophilizate can be reconstituted with water, saline, or another electrolyte to give a colloidal dispersion for intravenous or subcutaneous administration or can be formulated into a paste or filled into a soft gelatin or hard gelatin capsule for oral administration. Previously, camptothecins have not been administered subcutaneously because they are necrotic.
However, it appears that the lipid complex may sufficiently slow the release of water insoluble camptothecins that subcutaneous administration is possible.
While the invention will hereafter be described with respect to the preparation of lipid complexes and lyophiiizates of lipid complexes of 9-AC, those skilled in the art will appreciate that the methods taught herein are also applicable to the preparation of lipid complexes and lyophilizates of other camptothecins which are considered water insoluble such as cam~tothecin itself.
In accordance with the present invention, a lyophilizate of a phospholipid complex of 9-AC is prepared which can be reconstituted with pharmaceutically acceptable aqueous diluent such as water for injection and which, in comparison to solutions of 9-AC in dimethylacetamide, is less toxic, more stable and particularly importantly, its formulation and administration are not limited by the , solubility of 9-AC.
In accordance with the invention, the lyophiiizate , is prepared by a process comprising the steps of preparing a solution of 9-AC in a first organic solvent, preparing a solution of a phospholipid ~__ a second organic solvent, mixing the phospholipid solution and the 9-AC solution, adding water to the mi~sced solutions to cause formation cf a lipid complex of 9-AC, removing the first and second orcranic solvents to provide a dispersion of the lipid complex ~n water as an aqueous phase, dissolving a pharmaceutically acceptable excipient in the aqueous phase of the dispersion, and lyophilizing the dispersion of the lipid complex to form a lyophilizate. This lyophilizate forms a colloidal dispersion when reconstituted with a physiologically acceptable aqueous diluent.
In accordance with a preferred embodiment ef the present invention, the lyophilizate is prepared by a method comprising the steps of forming a concentrated solution of 9-AC in dimethyl sulfoxide, forming a concentrated solution of the phospholipids DMPC and DMPG in chloroform, mixing the phospholipid solution and the 9-AC solution, adding an aqueous solution such as water for injection to form the lipid complex of 9-AC and provide a dispersion of the 'ipid complex in water as an aqueous phase, sparging and diafiltering the dispersion to remove the solvents, red~;cing the particle size of the dispersion of the lipid complex, adding an aqueous solution of mannitol as a pharmaceutically acceptabl' lyophilization excipient to the dispersion, and lyophilizing, wherein a iyophilizate is obtained which :aon reconstit:a ing with water provides a colloidal dispersicn of a 9-AC lipid complex.
In accordance with a further embodiment cf the present invention, a lyophilized composition containing :-aminecamptothecin or another water insoluble camptcthecin is provided hhich comprises 9-AC, a phospholipid and a pharmaceutically acceptable lyophilization excipient.
Detailed Description of the Invention The term "water insoluble camptothecin" as used herein means camptothecin or another pentacyclic alkaloid having the aforementioned characteristic fused 5-ring system of camptothecin having a solubility in water which is less than .O1 mg/ml at 23°C.
The term "lipid complex" is an art recognized term. Lipid complexes are characterized by a noncovalent bond between the lipid and the camptothecin which is observed as a phase change in differential scanning calorimetry.
The term "pharmaceutically acceptable aqueous diluent" as used herein refers to water for injection, saline, and other known aqueous vehicles.
The term "lyophilization excipient" refers to a substance which is added to a solution prior to lyophilization to enhance characteristics such as the color, texture, strength, and volume cf the cake. Sxamples of lyophilization excipients are provided below 9-AC has been prepared by reduction of 9-nitrocamptothecin with a reducing agent such as a combination of tin or iron with a mineral acid. (See U.S.
Patent 4,604,463 to Miyasaka et al.) The preparation of 9-amino-20(S)- camptothecin and ~-amino-20(R,S)-camptothecin is described in U.S. Patents 5,106,742 and ,225,404. Other water insoluble camptothecins are known it the art and their preparation is described in the literature. , 9-Amino-20(S)-camptothecin has a chemical formula of C2o H,., N3 OQ and the structure:
o' A B CAN /
11 ~13 ~ 2 3 t5 T_n accordance with the invention a concentrated solution ef 9-AC in an organic solvent .s prepared. The most typical example of the solvent used to prepare this solution is dimethyl sulfoxide (DMSG). However, other organic solvents such as dimethyl formamide can be used. Useful solvents must form stable solutions wit:: the camptothecin analogue, e.g., the solvent must not interact with, destabilize, and/or deactivate the drug. In addition, the solubility cf the camptothecin analogue in the solvent must also be high enough that the camptothecin analogue can be dissolved ~r. amounts sufficiently high to form commercially useful quantities cf the lipid complex and the solvent should be capable of being removed easily from an aqueous dispersion c~ the lipid comr~iex as described hereinafter.
Preferably, a solution having a concentration of about 5 to SO mg/ml, preferably about 20 to 40 mg/rl and most preferably ~0 mg/ml camptothecin is used. The concentratior.
may vary depending upon the nature of the solvent and temperature, but it is important to use a concentrated solution o~ the camptothecin ir. preparing the liquid/camptothecin complex. "'his mini;tizes the amount of solvent that must be removed ~aLer In t-:e prC a c ss, anti also assists in forcing the camptotheci: out oT solution and into liQUid/camDtothecin complex formation with the ~ipid with tine addition c~ water.
The organic solvent used to prepare tha solution of the phospholipids should meet similar requirements to those outlined for the camptothecin solvent. It must be compatible with the phospholipids and not destabilize them or the camptothecins. In addition, the lipids should be soluble enough in the solvent so as to be able to introduce enough of the lipid to form the complex yet minimize the amount of solvent that must be removed later. A solvent which can be readily removed from the dispersion of the lipid complex is most preferred. The solvent most typically used to prepare this solution is chloroform or methylene chloride. Typically the concentration of this phospholipid solution will range from about 10 to 250 mg/ml.
Phospholipids are amphipathic in nature, i.e., the molecules have a hydrophobic tail such as a long chain hydrocarbon, and a hydrophilic head. In an aqueous medium, such as water or saline, the tails align with each other, away From the aqueous molecules, while the heads face outward into the aqueous phase. It is this nature of the phOSphOlipids that makes them very useful for formulating highly insoluble drugs like 9-AC.
The phospholipids used in tine invention are selected such that their phase transition temperature is about equal to or below the body temperature or 37~C and the complex releases the drug in the body. Representative examples of useful phospholipids include synthetic phospholipids DMPC, DMPG, dipalmitoylphosphatidylcholine (DPPC;, dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylcholine (DSPC), or distearoylphos-phatidylglycerol (DSPG), or a combination thereof. Other examples of phospholipids can be found in the CRC Handbook of Lipid Bilavers by Marsh, M.A., CRC Press (1990). When _g_ W O 96!11669 PCTIUS95I14105 s DMPC and DMPG are used in a ratio of DMPC to DMPG of about ' 7:3 they mimic the cell membrane.
The lipid solution is added to the 9-AC solution such teat the weight ratio of the 9-AC to lipid is about 1:80 to 1:5, preferably about 1:80 to 1:10, more preferably, about 1:60 to 1:10 and still more preferably about 1:45 to 1:25.
In some applications, it has been found desirable to add cholesterol or its hemisuccinate derivatives to the lipid complex. The cholesterol is believed to cause the biiayers to pack more closely and thereby slow the release of the drug. This approach may be particularly desirable with subcutaneous formulations where severe necrosis can result if the drug is delivered too auickly. The cholesterol is added to the phospholipid solution. The cholesterol may be used in an amount of about 0.5 to 15 parts per 100 parts phospholipid.
Once the solvent solutions of the lipids and camptcthecin have been mixed, water or an aqueous solution is added rapidly and with stirring for several minutes tc the mixture. Addition cf the water is believed to cause the 9-AC and the lipid to come out cf their respective solvent sol~:tions and complex with each other. The Ovate- .s preferably added in an amount such that the 9-AC is prese.~.t in an amount of about .05mg to 0.5mg per 100m1 water. It is desirable to limit the amount of water to minimize the amount of water which must be removed during the lyophilization process. Higher amounts of water are undesirable because they increase the amount of water that must be removed d~,~ring the subseauent lyophiiization process. It is believed complexat~c_~_ may be complete in about 3C minutes. However, it is desirable to stir the _g_ WO 96/11669 PCT/US95/141~5 dispersion for about one hour to insure complexation is complete.
The lipid complex dispersion described above, is treated to remove the solvents. Any of a variety of techniques can be used for this purpose. For example, it has been found that the chloroform can be removed if the dispersion is sparged with a gas such as nitrogen. A
diafiltration process (also known as a tangential flow filtration process) is used to remove the DMSO. A cartridge of hollow fiber tubes having a pore size of 5 to 150 kiiodaltons can be used. A diafiltration cartridge is available from A/G Technology Corporation cf Needham, Massachusetts under the tradename XPRESS, which can be used.
Other techniques which can be used include centrifugation.
A pharmaceutically acceptable iyophilization excipient is dissolved in the aqueous phase of the dispersion. Mannitol is typically used as the excipient but other excipients which do not interact with the drug or the lipid complex may be used. Sodium or potassium phosphate, citric acid, tartaric acid, gelatin, and carbohydrates such as mannitol, lactose, dextrose, dextran, hetastarch, etc.
are common examples of excipients which are also believed ~..
be useful herein. The excipients can be used alone or in combination to provide a cake of good quality which readily disperses in water upon reconstitution.
The excipients are typically added to the dispersion as solutions in water. Again, it is desirable to use concentrated solutions to minimize the amount of water .
for removal by lyophilization. The amount of the excipient is adjusted in a manner that is well known in the art to provide a cake which does not crack or shrink and is porous so that it readily dissolves and has a good appearance.
Mannitol has been found to be useful. Mannitoi is added tc WO 96/11669 ~ PCT/US95/14105 j the dispersion as a solution having a concentration of aboit to 150 g/ml. Mannitol is added in ar_ amount of about 1 to j 100 parts by weight per 1 part 9-AC.
After removing the solvents and adding the excipient, the dispersion is passed through a homogenizes (e.g.,. a Tekmar rotor/stator homogenizes, Model T25, or a microfluidics submerged jet homogenizes, Model M110Y). As a general rule, the smaller the particle size of the dispersion, the faster the formulation can be dried during 'i the lyophilization cycle. A dispersion having a particle size distribution ranging from about 10 to 500nm and v averaging about 250nm has been found to be satisfactory for j lyophilization. The optimum particle size may vary depending-on the mode of administration.
A typical lyophilization cycle useful in accordance with the present invention is provided below.
i ~ The cycle may be varied depending upon the equipment and facilities available in a manner well known in the art.
The homogenized formulation can be poured into vials of a 5 to 50m1 nominal volume. ':he vials are placed .. into a lyophilization chamber at about = °C. The vial size will usually be selected such that eacvial contains a single dosage cf the 9-AC: The temperature of the chambe r is reduced to -30°C over a period of one hour after which the temperature is maintained at -30°C for about four hours.
The pressure in the iyophilization.chamber is then reduced to 200-250 microns of pressure for the remainder of the cycle.. After reducing the pressure in the chamber, the temperature is ramped up to +25°C over a period of fifteen hours and the product is held at 25°C _:,= five hours. The temperature then is ramped up to +40°C over a period of 20 minutes and held at 40°C for two hours. The lyophilized * trademark -11-product preferably has a final moisture content of less than about 5o and typically about 1 to 2%. ' For intravenous or subcutaneous administration, the lyophilizate can be reconstituted using aqueous vehicles such as water, saline or another electrolyte. The lyophilized product with the addition of water provides a colloidal dispersion of the lipid complex in an aqueous solution of the excipient. Neither the 9-AC nor the lipids are soluble in water. A colloidal dispersion consists of at least two discrete phases. The first is a dispersed or internal phase. The second is a continuous or external phase. Systems in the colloidal state contain one or more substances that have at least one dimension in the range of 10-100A to a few microns. See pp. 272-4 in Chapter 19, Disperse Systems, Reminaton's Pharmaceutical Sciences, 18th Edition, 1990, Mack Publishing Company, Easton, PA 18042.
In the colloidal dispersions of the present invention, the dispersed cr internal phase comprises particles of 9-AC
lipid complex having a particle size in the range of lOnm to 1000nm. In selecting the aqueous vehicle, it is recommended to use one having a specific gravity about equal to the lipid complex (est. 1.09gjcc; to minimize the tendency for the dispersion to separate. The lyophilizate of the lipid complex can be reconstituted with water, saline, or another pharmaceutically acceptable aqueous diluent for intravenous administration. Upon reconstitution a dispersion is obtained which is suitable for injection. The lyophilizate can also be administered orally as an aqueous dispersion or , as a paste. While camptothecins are not generally administered subcutaneousl,r because they cause necrosis, it has been observed that the lipid slows the release of the camptothecin into the tissue making it potentially feasible to administer the lipid complex subcutaneousiy.
WO 96f11669 PCTlLIS9S/I4I05 For oral administration, the lyophilizate can be reconstituted tJ form an oral dispersion or formu~.ated into a paste. Alternatively, the lyophilizate can be filled into a soft gelatin capsule for oral administration.
Suitable dosages for 9-AC and camptothecins range from about 35 to 250 mg/m~/hour. The drug is preferably administered as a continuous infusion over 3 to 21 days using programmable continuous infusion ambulatory pump. It is anticipated that the drug will be administered with granulocyte colony stimulating factor (GCSF).
While it is contemplated herein that the lipid/camptothecin complex will be lyophilized to enhance its stability, it will be appreciated that the lipid/camptothecin complex is pharmaceutically active and can be formulated into a dosage form for oral, intravenous or subcutaneous administration without lyophilization.
Formulation aids such as antibacterials and antioxidants can be used to enhance the stability of the complex.
The invention will now be described in more detail with reference to the following non-limiting examples.
Examples ~-8 Lyophilizates of 9-AC were prepared using solvents, phospholipids and excipients in the amounts shown in Table 1. In each case, solutions were prepared by dissolving 9-AC in the amounts shown in DMSO, and, dissolving DMPC and DMPG in chloroform. The solutions were mixed and water for injection was added. The resulting dispersions were stirred for about one hour and cloroform was removed by sparging with nitrogen for about 95 min.
DMSO was removed by centrifuging the dispersion, removing the supernatant, and re-suspendir~g the "plug" in water for .E:':.:.
injection. tIn some examples, Millipore or AG Techno~ogy tangential flow filters were used to remove DMSO.) An aqueous solution of the excipient was then.
added to the dispersion and the dispersion was homogenized *.
>,;~ using a ULTRA TURREX homogenizes operating at approximately.
10,000 rpm, and room temperature. The homogenates were then lyophilized using the protocol described above or a similar process. , .
i --i Facamnie 1 2 3 4 ) 6 7 8 Na.
9 10.4 0.4 0.4 0.4 0.4 0.4 0.2 0.2 AC ma me mg mo mg mg mg ma -DMSO 0.01 0.01 0.01 8.7 '8.7 to 5 ml 5 ml ml ml ml ml ml ml - DMPC 126 24.2 3.0 22.4 22.4 22.4 11.2 8 ma~
~ ma ma mg ma m ma ma~
:-.,.-. DMPG _ 10.4 1.25 . 9.6 9.6 4.9.mg8 ma 11:3, ma mg 9.6 mg ma.
mg ma - Chloroform0.25 0_25 0.31 0.2 0.2 0.2 O.l 0.1 ml ml ml :~1 ml ml mg ml Mannitol 15.1 14.55 14 ma 15 15 20 120 20 ma ma ma ma ma ma ma ~ , Sodium 8.9 V8.9 mg' mg Citrate .. Cil_ = ~ 14.7 1y.7 .- ACid ma ~a WFI -Lyochilizate~0.4 10.4 0.4 0.2 ~0.2 12 1 ma/
mg/ mg/ mg/ mgl mg/ mg/
vial vial vial vial , vial vial vial The. lydphilizate obtained in Example 1 was placed on - stability testing at -4°C, 27°C, and 37°C. The initial assay showed~0.299 mg 9-AC-per,vial. the.results of the study to date are shown in Table 2.
* trademark WO 96/11669 PCTliIS95/I4I05 r Percent Initial Assav 4'C 27'C 37'C
1 week ~ 99.3 97.7 99.7 2 weeks ~ 100.3 103.3 104.3 1 month ~ 103.7 97.7 102.3 2 months ~ 101 102.3 102.3 months ~ 106.4 105.7 104.7 7 months I 106.4 104.2 ---The results of the study show no evidence of deterioration in the lyophilizate of the lipid/camptcthecin complex.
Example 9 Studies in animals have shown that the lyophilizates of 9-AC of the present invention exhibit antitumor activity in vivo. Colloidal dispersions of the lyophilizate were prepared by dispersing 1 mg cf the lyophilizate cf Example 6 above in 10 mi of water for injection. The dispersions were evaluated against the HT29 human colon tumor xenograft in female athymic mice. Both intravenous and oral administrations were used. Tumor xenografts were allowed to grow to abou~ 200 mg before initiation of chemotherapy. The size c. the tumor was determined based on tumor volume. Treatment comparisons were based on time (days) to three tumor doublings (TTTD).
The results are summarized in the Table 3. A high level of citrate was present in the formulation curing the first course of treatment and resulted in a nigh incidence of vehicle-related mortality at the highes~ I.v. dose levels.
The formulation was orally active on a Q2Dxl4 schedule with no toxicity.
I.V. and P.O. Activity of 9-AC vs. HT29 Xeno r CDF
Schedule mg/kg Dose TTTD Deaths 0 26.8 0 I.V. Bolus 1.8 34.2 0 Q4Dx6 2.7 38.9 2a (6 doses) ~ 4.0 ~ 25.3 ~ 6a I.V. Bolus 0 26.3 0 Qdx4 0.9 35.4 0 Days 12,19,26 1.33 46.4 2b 12 doses 2.0 50.9 7b 0 27.0 0 P.~. 0.67 35.6 0 1.0 33.7 0 Q2Dx14 1.5 42.8 0 CDF = Collidal dispersion formulation (lipid formulation); TTTD = Time (days) to 2 X doubling of tumor size Start Treatment Day 12 for I.V., Day 13 for P.O.; N =
10/group First dose or CDF contained citrate buffer. Most deaths attributed to vehicle. Second and remaining doses were citrate-free First course of DMSO/intralipid containing citrate buffer. Most deaths attributed to vehicle. Second and third courses were citrate-free 1 partial regression SUBSTITUTE SHEET (RULE 2G) i Having described the invention in detail and by reference to preferred embodiments thereof, it will be apparent that modifications and variations are possible ~;- ~ without departing from the scope of the invention defined in the appended claims.
. -16/i-SUBSTITUTE SHEET (RULE 26)
Claims (18)
1. A pharmaceutical composition comprising a lyophilizate of a non-liposomal complex of a water insoluble camptothecin and a phospholipid and a pharmaceutically acceptable lyophilization excipient, wherein the phase transition temperature of the phospholipid is about equal to or below body temperature of 37°C; and wherein the weight ratio of the water insoluble camptothecin to the phospholipid ranges from 1:80 to 1:5.
2. The composition of claim 1 wherein the camptothecin and the phospholid are present as a lipid complex.
3. The composition of claim 2 wherein said composition forms a colloidal dispersion when reconstituted with a physiologically acceptable aqueous diluent, wherein the particles in the colloidal dispersion have a particle size ranging from 10 nm to 1000 nm.
4. The composition of claim 2 wherein said phospholipid is selected from the group consisting of dimyristoylphosphatidyl choline, dimyristoylphosphatidyl glycerol, dipalmitoyliphosphatidyl choline, dipalmitoylphophatidyl glycerol, distearoylphosphatidyl choline, distearoylphosphatidyl glycerol, and any combination thereof.
5. The composition of claim 2 wherein said phospholipid is a mixture of dimyristoylphosphatidyl choline and dimyriscoylphosphatidyl glycerol.
6. The composition of claim 5 wherein said dimyristoylphosphatidyl choline is present in a weight ratio to said dimyristoylphosphatidyl glycerol of 7:3.
7. The composition of claim 5 wherein the weight ratio of said camptothecin to said phospholipid in said composition is from 1:60 to 1:10.
8. The composition of claim 5 wherein said excipient is mannitol.
9. The composition of claim 5 wherein said water insoluble camptothecin is 9-aminocamptothecin.
10. The composition of claim 9 wherein the 9-aminocamptothecin is 9-amino-20-(S)-camptothecin.
11. The composition of claim 9 wherein the 9-aminocamptothecin is 9-amino-20-(R,S)-camptothecin.
12. The composition of claim 1 wherein the composition is suitable for intravenous administration.
13. The composition of claim 1 wherein the composition is suitable for oral administration.
14. The composition of claim 1 wherein the composition is suitable for subcutaneous administration.
15. The composition of claim 5 wherein the composition additionally contains cholesterol.
16. The composition of claim 3 wherein the diluent has a specific gravity approximately equal to the lipid complex.
17. The composition of claim 1, wherein the camptothecin and its analogs is selected from the group consisting of 7-ethyl-[14-(1-piperidino)-1-piperidine]
carbonyloxycamptothecin (CPT-11); 20-(S)-camptothecin; 10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)dione monohydrochloride(topotecan hydrochloride); and 9-amino-20(S)-camptothecin.
carbonyloxycamptothecin (CPT-11); 20-(S)-camptothecin; 10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)dione monohydrochloride(topotecan hydrochloride); and 9-amino-20(S)-camptothecin.
18. The composition of claim 1, wherein the camptothecin analog is 7-ethyl-[14-(1-piperidino)-1-piperidine] carbonyloxycamptothecin (CPT-11).
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US32308194A | 1994-10-14 | 1994-10-14 | |
| US08/323,081 | 1994-10-14 | ||
| PCT/US1995/014105 WO1996011669A1 (en) | 1994-10-14 | 1995-10-11 | Lyophilizate of lipid complex of water insoluble camptothecins |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2202531A1 CA2202531A1 (en) | 1996-04-25 |
| CA2202531C true CA2202531C (en) | 2005-05-03 |
Family
ID=23257667
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002202531A Expired - Fee Related CA2202531C (en) | 1994-10-14 | 1995-10-11 | Lyophilizate of lipid complex of water insoluble camptothecins |
Country Status (18)
| Country | Link |
|---|---|
| US (1) | US6548071B1 (en) |
| EP (1) | EP0785772A4 (en) |
| JP (2) | JPH10507454A (en) |
| KR (1) | KR100387744B1 (en) |
| CN (1) | CN1138543C (en) |
| AU (1) | AU689983B2 (en) |
| CA (1) | CA2202531C (en) |
| HU (1) | HUT77975A (en) |
| IL (1) | IL115099A (en) |
| MY (1) | MY113203A (en) |
| NO (1) | NO323582B1 (en) |
| NZ (1) | NZ296389A (en) |
| PL (1) | PL183040B1 (en) |
| RU (1) | RU2169555C2 (en) |
| TW (1) | TW450811B (en) |
| UA (1) | UA64692C2 (en) |
| WO (1) | WO1996011669A1 (en) |
| ZA (1) | ZA957860B (en) |
Families Citing this family (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19758157A1 (en) | 1997-03-27 | 1998-10-01 | Sueddeutsche Kalkstickstoff | Homogeneous anhydrous glycero-phospholipid formulation |
| GB9925127D0 (en) * | 1999-10-22 | 1999-12-22 | Pharmacia & Upjohn Spa | Oral formulations for anti-tumor compounds |
| US20040009229A1 (en) * | 2000-01-05 | 2004-01-15 | Unger Evan Charles | Stabilized nanoparticle formulations of camptotheca derivatives |
| BR0115260A (en) | 2000-11-09 | 2005-08-16 | Neopharm Inc | Sn-38 lipid complexes and their uses in treating diseases |
| WO2003030864A1 (en) | 2001-05-29 | 2003-04-17 | Neopharm, Inc. | Liposomal formulation of irinotecan |
| DE10234784A1 (en) * | 2002-07-30 | 2004-02-19 | Günenthal GmbH | Pharmaceutical dosage form that can be administered intravenously |
| US7838034B2 (en) | 2002-07-30 | 2010-11-23 | Grunenthal Gmbh | Intravenous pharmaceutical form of administration |
| WO2004035032A2 (en) * | 2002-08-20 | 2004-04-29 | Neopharm, Inc. | Pharmaceutical formulations of camptothecine derivatives |
| US20060030578A1 (en) * | 2002-08-20 | 2006-02-09 | Neopharm, Inc. | Pharmaceutically active lipid based formulation of irinotecan |
| US20040247624A1 (en) * | 2003-06-05 | 2004-12-09 | Unger Evan Charles | Methods of making pharmaceutical formulations for the delivery of drugs having low aqueous solubility |
| JP2007522085A (en) * | 2003-06-27 | 2007-08-09 | スミスクライン・ビーチャム・コーポレイション | Stabilized topotecan liposome compositions and methods |
| JP4433918B2 (en) * | 2004-07-15 | 2010-03-17 | コニカミノルタエムジー株式会社 | Image forming method |
| KR100651728B1 (en) * | 2004-11-10 | 2006-12-06 | 한국전자통신연구원 | Compound for an electronic device having a fixing group, an electronic device comprising the same and a method of manufacturing the same |
| CN1326525C (en) * | 2004-11-26 | 2007-07-18 | 复旦大学 | 10-hydroxy camptothecin long cyclic liposome and its freeze aried preparation |
| TW200727900A (en) * | 2005-07-27 | 2007-08-01 | Yakult Honsha Kk | Aqueous solution preparation containing camptothecins |
| CA2631243A1 (en) * | 2005-12-08 | 2007-06-14 | Wyeth | Liposomal compositions |
| CN101028251B (en) * | 2007-02-14 | 2010-05-19 | 四川大学 | Camptothecin derivative phospholipid complex lipid nanoparticle preparation and preparation method thereof |
| CN101721364B (en) * | 2009-11-10 | 2014-05-21 | 沈阳药科大学 | A kind of aqueous dispersion of camptothecin compound phospholipid complex and preparation method thereof |
| AU2013347990B2 (en) | 2012-11-20 | 2018-01-18 | Arbutus Biopharma Corp. | Improved method for the preparation of liposome encapsulated vincristine for therapeutic use |
| TWI678213B (en) | 2015-07-22 | 2019-12-01 | 美商史倍壯製藥公司 | A ready-to-use formulation for vincristine sulfate liposome injection |
| DE102018200520A1 (en) * | 2018-01-15 | 2019-07-18 | Robert Bosch Gmbh | A method for providing a solution of the substance in a microfluidic device |
Family Cites Families (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AT347054B (en) * | 1973-09-29 | 1978-12-11 | Takeda Chemical Industries Ltd | PROCESS FOR THE PRODUCTION OF NEW NONAPEPTIDAMIDE DERIVATIVES |
| GB1575343A (en) * | 1977-05-10 | 1980-09-17 | Ici Ltd | Method for preparing liposome compositions containing biologically active compounds |
| CH621479A5 (en) * | 1977-08-05 | 1981-02-13 | Battelle Memorial Institute | |
| US4610868A (en) | 1984-03-20 | 1986-09-09 | The Liposome Company, Inc. | Lipid matrix carriers for use in drug delivery systems |
| US5279811A (en) * | 1987-02-18 | 1994-01-18 | The Du Pont Merck Pharmaceutical Company | Ester-substituted diaminedithiols and radiolabeled complexes thereof |
| US5616334A (en) * | 1987-03-05 | 1997-04-01 | The Liposome Company, Inc. | Low toxicity drug-lipid systems |
| US5227380A (en) * | 1987-03-31 | 1993-07-13 | Research Triangle Institute | Pharmaceutical compositions and methods employing camptothecins |
| MX9203504A (en) * | 1988-04-20 | 1992-07-01 | Liposome Co Inc | AGENT COMPLEX: HIGH PROPORTION ACTIVE LIPID. |
| US5077057A (en) | 1989-04-05 | 1991-12-31 | The Regents Of The University Of California | Preparation of liposome and lipid complex compositions |
| US5389377A (en) | 1989-12-22 | 1995-02-14 | Molecular Bioquest, Inc. | Solid care therapeutic compositions and methods for making same |
| US5552156A (en) * | 1992-10-23 | 1996-09-03 | Ohio State University | Liposomal and micellular stabilization of camptothecin drugs |
| WO1994026253A1 (en) * | 1993-05-19 | 1994-11-24 | The Liposome Company, Inc. | Liposome having a multicomponent bilayer which contains a bioactive agent as an integral component of the bilayer |
| WO1995008986A1 (en) * | 1993-09-27 | 1995-04-06 | Smithkline Beecham Corporation | Camptothecin formulations |
| US5834012A (en) * | 1995-05-03 | 1998-11-10 | Roman Perez-Soler | Lipid complexed topoisomerase I inhibitors |
| US6090407A (en) * | 1997-09-23 | 2000-07-18 | Research Development Foundation | Small particle liposome aerosols for delivery of anti-cancer drugs |
-
1995
- 1995-08-29 IL IL11509995A patent/IL115099A/en not_active IP Right Cessation
- 1995-09-18 TW TW084109751A patent/TW450811B/en not_active IP Right Cessation
- 1995-09-18 ZA ZA957860A patent/ZA957860B/en unknown
- 1995-10-11 HU HU9800995A patent/HUT77975A/en unknown
- 1995-10-11 WO PCT/US1995/014105 patent/WO1996011669A1/en active IP Right Grant
- 1995-10-11 PL PL95319588A patent/PL183040B1/en not_active IP Right Cessation
- 1995-10-11 JP JP8513489A patent/JPH10507454A/en active Pending
- 1995-10-11 RU RU97107856/14A patent/RU2169555C2/en not_active IP Right Cessation
- 1995-10-11 EP EP95939009A patent/EP0785772A4/en not_active Withdrawn
- 1995-10-11 KR KR1019970702418A patent/KR100387744B1/en not_active Expired - Fee Related
- 1995-10-11 NZ NZ296389A patent/NZ296389A/en unknown
- 1995-10-11 CN CNB951956590A patent/CN1138543C/en not_active Expired - Fee Related
- 1995-10-11 CA CA002202531A patent/CA2202531C/en not_active Expired - Fee Related
- 1995-10-11 AU AU40185/95A patent/AU689983B2/en not_active Ceased
- 1995-10-12 MY MYPI95003048A patent/MY113203A/en unknown
- 1995-11-10 UA UA97041743A patent/UA64692C2/en unknown
-
1997
- 1997-04-11 NO NO19971668A patent/NO323582B1/en unknown
- 1997-06-10 US US08/871,558 patent/US6548071B1/en not_active Expired - Fee Related
-
2007
- 2007-09-19 JP JP2007242506A patent/JP2008024717A/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| UA64692C2 (en) | 2004-03-15 |
| PL319588A1 (en) | 1997-08-18 |
| TW450811B (en) | 2001-08-21 |
| AU689983B2 (en) | 1998-04-09 |
| KR970706792A (en) | 1997-12-01 |
| RU2169555C2 (en) | 2001-06-27 |
| KR100387744B1 (en) | 2003-11-28 |
| NO971668L (en) | 1997-04-11 |
| JPH10507454A (en) | 1998-07-21 |
| US6548071B1 (en) | 2003-04-15 |
| NO971668D0 (en) | 1997-04-11 |
| MY113203A (en) | 2001-12-31 |
| IL115099A0 (en) | 1995-12-08 |
| JP2008024717A (en) | 2008-02-07 |
| CN1138543C (en) | 2004-02-18 |
| CA2202531A1 (en) | 1996-04-25 |
| NZ296389A (en) | 1999-06-29 |
| WO1996011669A1 (en) | 1996-04-25 |
| EP0785772A1 (en) | 1997-07-30 |
| CN1160344A (en) | 1997-09-24 |
| HUT77975A (en) | 1999-01-28 |
| EP0785772A4 (en) | 2005-12-21 |
| AU4018595A (en) | 1996-05-06 |
| PL183040B1 (en) | 2002-05-31 |
| NO323582B1 (en) | 2007-06-11 |
| ZA957860B (en) | 1996-05-21 |
| IL115099A (en) | 1999-04-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2202531C (en) | Lyophilizate of lipid complex of water insoluble camptothecins | |
| US5707608A (en) | Methods of making liposomes containing hydro-monobenzoporphyrin photosensitizer | |
| US20050238706A1 (en) | Pharmaceutically active lipid based formulation of SN-38 | |
| US20060222694A1 (en) | Stabilized topotecan liposomal composition and methods | |
| KR100711315B1 (en) | Liposomal Camptothecin Formulation | |
| EP1030686A2 (en) | Lyophilizate of lipid complex of water insoluble porphyrins | |
| US8067432B2 (en) | Liposomal, ring-opened camptothecins with prolonged, site-specific delivery of active drug to solid tumors | |
| US20010010822A1 (en) | Lipid complexes and liposomes of highly insoluble platinum complexes | |
| US6740335B1 (en) | Liposomal camptothecin formulations | |
| US20020142048A1 (en) | Injectable pharmaceutical composition comprising coated particles of camptothecin | |
| US20020058643A1 (en) | Lyophilizate of lipid complex of water insoluble porphyrins | |
| US6497896B2 (en) | Method for administering camptothecins via injection of a pharmaceutical composition comprising microdroplets containing a camptothecin | |
| US20020150615A1 (en) | Injectable pharmaceutical composition comprising microdroplets of a camptothecin | |
| EP1365705A1 (en) | Injectable pharmaceutical composition comprising microparticles or microdroplets of camptothecin | |
| EP1227795B1 (en) | Lipid complex of alkycyclines | |
| WO1993023016A1 (en) | Liposomal formulations containing rifamycins |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |