WO2009114462A2

WO2009114462A2 - Local delivery of chemotherapeutic agents to skeletal metastases

Info

Publication number: WO2009114462A2
Application number: PCT/US2009/036512
Authority: WO
Inventors: John J. Barry; Robert K. Heck
Original assignee: Inmotion Musculoskeletal Institute
Priority date: 2008-03-11
Filing date: 2009-03-09
Publication date: 2009-09-17
Also published as: WO2009114462A3

Abstract

The present invention relates to the field of cancer treatment in anatomical structures such as the human skeleton. More specifically, the present invention relates to the local delivery of chemotherapeutic agents useful for the treatment and prevention of bone pain associated with skeletal metastases, the prevention of growth or progression of skeletal metastases, and the prevention and treatment of pathologic fractures caused by skeletal metastases. A method of treating bone in a patient suffering from skeletal metastases is also provided.

Description

TITLE OF THE INVENTION

Local Delivery of Chemotherapeutic Agents to Skeletal Metastases

INVENTORS John J. BARRY, Vienna, AUSTRIA (IE) ; and

Robert K. HECK, Memphis, Tennessee, USA (US).

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This Non-Provisional Patent Application claims the benefit of U.S. Provisional Patent Application No. 61/035,709, filed on 11 March 2008, and which is hereby incorporated by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT [0002] Not applicable.

THE NAMES OF THE PARTIES TO A JOINT RESEARCH AGREEMENT [0003] Not applicable.

INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON COMPACT DISC

[0004] Not applicable.

BACKGROUND OF THE INVENTION [0005] 1. Field of the Invention

[0006] The present invention relates to the field of cancer treatment. More specifically, the present invention relates to the local delivery of a chemotherapeutic agent useful for the treatment and prevention of bone pain associated with skeletal metastases, the prevention of growth or progression of skeletal metastases, and the prevention and treatment of pathologic fractures caused by skeletal metastases.

[0007] 2. Description of Related Art

[0008] Over 1.3 million cases of carcinoma are diagnosed in the United States each year. Approximately 50 to 80 % of the patients diagnosed with carcinoma will exhibit skeletal metastases at some point in the disease course. Skeletal metastasis means that cancer cells have spread from one organ or cellular structure into the bone structure of a patient.

[0009] With improved treatments, cancer patients are living longer. However, symptoms and medical complications associated with skeletal metastases can be intractable. Such complications extend from simple bone pain to osteolysis and bone fractures.

[0010] Patients who exhibit skeletal metastases have limited options for arresting tumor development on distal limbs. Current treatments use radiation or harmful chemicals dispersed throughout the body. These treatments are time consuming and costly.

[0011] In the case of radiation, a two-week course of radiotherapy is the current standard of care for skeletal metastases. The affected areas are subjected to a beam from an ionizing source causing irreversible damage to the cancerous cells. Unfortunately, along with damage to the cancerous cells there are various possible side effects to the patient, depending on the duration and site of the treatment. These can include damage to epithelial surfaces, swelling, fatigue, fibrosis, diarrhea, nausea, and hair loss. Bone metastases can also be refractory to the radiotherapy treatment.

[0012] In the case of chemical treatment, a chemotherapeutic solution may be administered either orally or intravenously. However, the chemicals may attack not only the tumor, but may also attack healthy systems of the patient. Chemical treatments are generally believed to offer poor pharmacokinetics, or drug delivery. The difficulties associated with oral delivery of complex anti-cancer formulations present an even greater obstacle when treating localized or specific indications such as bone metastases, as compared with systemic indications. [0013] A clinical need exists for a chemotherapeutic agent that can be used to target and treat specific skeletal metastatic sites while having reduced systemic side-effects. The local delivery of chemotherapeutic agents provides new possibilities for the physician's treatment of patients. Such treatments may extend beyond the skeletal structure and be used to treat metastases in other organs as well.

BRIEF SUMMARY OF THE INVENTION

[0014] According to one aspect, the present invention relates to a self-curing fibrin biomatrix. The fibrin matrix defines a natural bio-polymer that is sterilizable and has low toxicity. The fibrin matrix is resorbable, meaning that it is biologically degradable

(biodegradable; a resorbable composition). The resorbable fibrin matrix includes a cytotoxic drug for the local treatment of metastases. Skeletal metastases are described as an example. However, metastases on other organs may also be treated including metastases of the lungs, brain or liver. Any cytotoxic drug or chemotherapeutic agent may be used so long as it is soluble in a contrast agent. The taxane family of drugs is descried as an example. Any known or generic taxane may be used provided again that it is soluble in a contrast agent.

[0015] The composition also includes a contrast agent. An iodine-containing contrast agent is described as an example. The purpose of the contrast agent is to provide radio- opacity for the biomatrix as would be used during fluoroscopy. Inclusion of an iodine- containing contrast media to the fibrin matrix allows for accurate delivery and post surgical follow up of the composition using fluoroscopy.

[0016] In one embodiment, the resorbable composition comprises fibrinogen, thrombin, a contrast agent, and a chemotherapeutic agent. Preferably, the contrast agent contains iodine, and more preferably the contrast agent is selected from the group consisting of iodecol, iodixanol, iofratol, iogulamide, iohexol, ioxaglate, iomeprol, iopamidol, iopromide, iotrol and ioversol. The contrast agent may be present in an amount from about 100 mg/mL to about 800 mg/mL, preferably in an amount from about 150 mg/mL to about 600mg/mL, and more preferably in an amount from about 200 mg/mL to about 400 mg/mL. The chemotherapeutic agent is preferably a taxane, and more preferably is selected from the group consisting of paclitaxel, docetaxel, milataxel, ortataxel, and protaxel. The fibrinogen may be present in an amount from about 5 mg/mL to about 100 mg/mL, preferably in an amount from about 15 mg/mL to about 75 mg/mL, and more preferably in an amount from about 35 mg/mL to about 63 mg/mL. The thrombin may be present in an amount from about 1 IU/mL to about 1000 IU/mL, preferably in an amount from about 30 IU/mL to about 300 IU/mL, and more preferably in an amount from about 100 IU/mL to about 250 IU/mL. When combined, the fibrinogen and thrombin form fibrin, the combination yielding fibrin in an amount from about 10 mg/mL to about 200 mg/mL, preferably in an amount from about 30 mg/mL to about 150 mg/mL, and more preferably in an amount from about 75 mg/mL to about 115 mg/mL.

[0017] This invention also relates to a method of treating cancer in a mammal in need of such treatment, the method comprising administering to the mammal an effective amount of a resorbable composition comprising fibrinogen, thrombin, a contrast agent, and a chemotherapeutic agent. In one embodiment, the cancer is a bone cancer, preferably a skeletal metastasis. In another aspect, the mammal is a human. The administering may comprise placing the resorbable composition into a bone affected by said skeletal metastasis. Preferably, the contrast agent contains iodine, and more preferably the contrast agent is selected from the group consisting of iodecol, iodixanol, iofratol, iogulamide, iohexol, ioxaglate, iomeprol, iopamidol, iopromide, iotrol and ioversol. The contrast agent may be present in an amount from about 100 mg/mL to about 800 mg/mL, preferably in an amount from about 150 mg/mL to about 600mg/mL, and more preferably in an amount from about 200 mg/mL to about 400 mg/mL. The chemotherapeutic agent is preferably a taxane, and more preferably is selected from the group consisting of paclitaxel, docetaxel, milataxel, ortataxel, and protaxel. The fibrinogen may be present in an amount from about 5 mg/mL to about 100 mg/mL, preferably in an amount from about 15 mg/mL to about 75 mg/mL, and more preferably in an amount from about 35 mg/mL to about 63 mg/mL. The thrombin may be present in an amount from about 1 IU/mL to about 1000 IU/mL, preferably in an amount from about 30 IU/mL to about 300 IU/mL, and more preferably in an amount from about 100 IU/mL to about 250 IU/mL. When combined, the fibrinogen and thrombin form fibrin, the combination yielding fibrin in an amount from about 10 mg/mL to about 200 mg/mL, preferably in an amount from about 30 mg/mL to about 150 mg/mL, and more preferably in an amount from about 75 mg/mL to about 115 mg/mL. [0018] The present invention also relates to a method of making a resorbable composition comprising fibrinogen, thrombin, a contrast agent, and a chemotherapeutic agent, the method comprising: reconstituting said fibrinogen, to make a fibrinogen solution; reconstituting said thrombin, to make a thrombin solution; mixing a therapeutically appropriate amount of the chemotherapeutic agent with the contrast agent, to make a chemotherapeutic agent solution; mixing the thrombin solution with the chemotherapeutic agent solution to make a working thrombin solution; and mixing the working thrombin solution with the fibrinogen solution. Preferably, the contrast agent contains iodine, and more preferably the contrast agent is selected from the group consisting of iodecol, iodixanol, iofratol, iogulamide, iohexol, ioxaglate, iomeprol, iopamidol, iopromide, iotrol and ioversol. The contrast agent may be present in an amount from about 100 mg/mL to about 800 mg/mL, preferably in an amount from about 150 mg/mL to about 600mg/mL, and more preferably in an amount from about 200 mg/mL to about 400 mg/mL. The chemotherapeutic agent is preferably a taxane, and more preferably is selected from the group consisting of paclitaxel, docetaxel, milataxel, ortataxel, and protaxel. The fibrinogen may be present in an amount from about 5 mg/mL to about 100 mg/mL, preferably in an amount from about 15 mg/mL to about 75 mg/mL, and more preferably in an amount from about 35 mg/mL to about 63 mg/mL. The thrombin may be present in an amount from about 1 IU/mL to about 1000 IU/mL, preferably in an amount from about 30 IU/mL to about 300 IU/mL, and more preferably in an amount from about 100 IU/mL to about 250 IU/mL. When combined, the fibrinogen and thrombin form fibrin, the combination yielding fibrin in an amount from about 10 mg/mL to about 200 mg/mL, preferably in an amount from about 30 mg/mL to about 150 mg/mL, and more preferably in an amount from about 75 mg/mL to about 115 mg/mL.

DETAILED DESCRIPTION OF THE INVENTION

[0019] Before the subject invention is further described, it is to be understood that the invention is not limited to the particular embodiments of the invention described below, as variations of the particular embodiments may be made and still fall within the scope of the appended claims. It is also to be understood that the terminology employed is for the purpose of describing particular embodiments, and is not intended to be limiting. Instead, the scope of the present invention will be established by the appended claims.

[0020] In this specification and the appended claims, the singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this invention belongs. [0021] As used herein, the term "contrast agent" includes any contrast agent usable in imaging applications known in the art such as fluoroscopy.

[0022] The term "taxane" means any type of mitotic inhibitor derived from diterpenes produced by the plants of the genus Taxus (yews). The term "taxane" also includes synthesized materials of natural diterpenes. Examples include Paclitaxel (Taxol), BMSl 84476 (7-methylthiomethylpaclitaxel); BMS 188797; BMS 275183; CYC-3204 (a penetratin-Paclitaxel conjugate); Taxoprexin; DJ-927; Docetaxel (Taxotere); XRP9881 (RPR-109881A); XRP6258 (RPRl 12658); Milataxel; MST 997; MBT-206; NBT-287; ortataxel; Protax-3; PG-TXL; PNU- 166945; 106258; BMS-188797; 109881; BAY 598862 (IDN 5109; semisynthetic taxane); Protaxel, and MAC-321 (Taxalog).

[0023] As used herein, the term "iodine-containing" includes all compounds which contain at least one iodine atom and/or iodine ion, bonded either physically or chemically, for example covalenfly or coordinatively. Examples of suitable iodine-containing organic compounds include iodecol, iodixanol, iofratol, iogulamide, iomeprol, iopamidol, iopromide, iotrol and ioversol.

[0024] The term "thrombin" includes not only thrombin per se, but also any gelation- inducing or clot-inducing agent. An example is a physiologically acceptable alkaline buffer system.

[0025] As used herein, the term "fibrinogen" includes not only fibrinogen per se, but also any clot-forming substance made of fibrinogen, such as clot-forming derivatives of fibrinogen, for example "fibrinl."

[0026] The term "fibrin" means a fully or partly coagulated fibrinogen. In addition, "fibrin" (also called "Factor Ia") means or refers to any mixture of fibrin and fibrinogen which may occur during formation of fibrin from fibrinogen using thrombin. Fibrin is a fibrous protein involved in the clotting of blood, and is non-globular. It is made from fibrinogen. Fibrinogen is converted to fibrin by the serine protease thrombin. Fibrin may then be cross-linked by factor XIII to form a clot.

[0027] The term "clotted" means, for example, a gel comprising fibrin and includes any kind of coagulation state known in the art. [0028] "Fibrin matrix" means any combination of proteins from the blood clotting cascade that, when combined, form a three dimensional structure or matrix. This matrix has a porous structure permitting the moving of cells and fluids through the structure. It may also contain ancillary protein such as fibronectin which facilitate cell attachment and spreading on the structure.

[0029] As used herein, the term "patient" means a subject suffering from a disorder such as metastasis, including bone metastasis, and includes mammals such as human beings.

[0030] The present invention provides a fibrin composition for the treatment of metastases. The composition is self-curing, meaning that it clots into a fibrin matrix. When cured, the fibrin matrix defines a natural bio-polymer that is sterilizable and has low toxicity. The fibrin matrix is resorbable, meaning that it is biologically degradable. In addition, the fibrin matrix has substantially no exothermicity upon curing. In some embodiments the fibrin matrix may exhibit mechanical properties typically seen in elastomers so that the cured composition is mechanically superior to fibrin alone.

[0031] The composition is made of fibrinogen, a soluble plasma glycoprotein synthesized by the liver. As the precursor to fibrin, fibrinogen is the principal protein of vertebrate blood clotting. The protein may be polymerized to form a mesh or matrix useful for a haemostatic clot. According to one example of the present composition, the concentration of fibrinogen in the composition ranges from about 5 to about 100 mg/mL, such as from about 15 to 75 mg/mL or from about 35 to 63 mg/mL in the final clotted composition. In one example, the fibrinogen component is a freeze-dried fibrinogen powder reconstituted with an aprotinin solution to a total clottable protein concentration of about 91 mg/mL.

[0032] The fibrinogen component may comprise one or more of extracellular matrix proteins. Examples include fibronectin, cellular associated proteins, and other plasma derived proteins, for example blood clotting factor XIII (FXIII), proteases, protease inhibitors, and mixtures thereof. The fibrinogen component may also include any additive which is comprised in the state of the art for scientific and/or commercially available fibrinogen compositions, for example commercially available fibrinogen solutions.

[0033] In one aspect, the composition for the treatment of metastases also comprises thrombin. Thrombin is a serine protease enzyme that converts soluble fibrinogen into insoluble strands of fibrin. Thrombin is also a coagulation protein that may be extracted from available blood supplies. The thrombin is mixed with the fibrinogen to form the fibrin matrix.

[0034] There is no specific limitation in respect to the amount of thrombin to be used in the composition. In one example, the amount of thrombin is about 1 IU/mL to about 50 IU/mL in the final clotted composition, such as about 30 IU/mL. In one example the thrombin is a freeze-dried thrombin powder, reconstituted with 5 mL of thrombin buffer to a concentration of 500 IU/mL. The thrombin buffer may be 40 mM CaCl₂ in H₂O. In another example, 1000 IU of freeze-dried thrombin powder is reconstituted in 2 mL of a solution containing 85.4 ng/mL paclitaxel (PTX), 0.005% Cremaphor EL, 0.005% Ethanol, 99.9% Optiprep, and 4OmM calcium chloride, yielding a thrombin solution (500 IU/mL thrombin) .

[0035] The fibrin matrix includes any ratio of fibrinogen / thrombin and any grade of gelation and/or clotting as long as it has no negative impact on the final composition injected into the non-mineralized or hollow portion of a patient's bone. In one aspect, a 1:1 ratio of fibrinogen and thrombin is used. The amount of fibrin formed from the combined fibrinogen and thrombin ranges, for example, from about 10 to about 200 mg/mL, such as from about 30 to about 150 mg/mL or from about 75 to about 115 mg/mL. These figures are about twice the figures for the fibrinogen concentrations provided above.

[0036] The fibrin matrix is loaded with a cytotoxic agent or chemotherapeutic drug.

Preferably, a member of the taxane family of drugs is described as an example. Taxanes are compounds that prevent cell replication by stabilizing cell microtubules. Microtubules are needed for cell division, and taxanes are known to inhibit cell division.

[0037] Any known or generic taxane may be used. Specific examples of taxanes are paclitaxel and docetaxel. Paclitaxel was originally derived as an extract from the Pacific yew tree, Taxus brevifolia. Paclitaxel may be purchased under the trademark Taxol™. (Bristol-Myers Squibb, New York, New York). Docetaxel is a semi-synthetic analogue of paclitaxel. Docetaxel may be purchased under the trademark Taxotere™. Both paclitaxel and docetaxel are understood to prevent cell replication.

[0038] Taxanes have been used in connection with oral and intravenous delivery for the treatment of various forms of cancer including breast, prostate, colorectal, lung, and ovarian cancer. Because taxanes tend to be hydrophobic, taxane-containing drugs are usually delivered in an excipient containing castor oil. For example, the excipient may be Cremaphor EL (a polyoxyethylated castor oil) coupled with dehydrated ethanol. Several reports have linked the excipient to alterations in the pharmacokinetic profiles of both paclitaxel and docetaxel. Therefore, it is desirable to use the cytostatic taxane agent without the polyoxyethylated castor oil excipient.

[0039] It is desirable to deliver the taxane-containing composition locally for the treatment of skeletal metastases. More specifically, the taxane-loaded, fibrin-based matrix may be used as a chemotherapeutic agent for local delivery. The composition may also be used for the local treatment of other metastasis forms such as lung or brain metastases.

[0040] The therapeutic composition also includes a contrast agent. The contrast agent provides radio-opacity for the fibrin matrix as is desirable for fluoroscopy. Preferably, the contrast media includes iodine. The use of an iodine-containing contrast media in the composition allows for accurate delivery and post-surgical follow up of the composition using fluoroscopy. In one embodiment of the present invention, the suitable contrast agent has a low osmolality and allows fibrin assembly to occur at an appropriate extent.

[0041] The contrast agent is made of at least one iodine-containing organic compound. As used herein, the term "iodine-containing organic compound" includes all compounds which contain at least one iodine atom and/or iodine ion, bonded either physically or chemically, for example covalently or coordinatively. Examples of suitable iodine-containing organic compounds include iodecol, iodixanol, iofratol, iogulamide, iohexol, iomeprol, iopamidol, iopromide, iotrol, ioversol, and ioxaglate.

[0042] It is believed that iodine-containing contrast media function well as vehicles for taxanes. In this respect, taxane has been found to be soluble in the iodine-containing contrast agent. Thus, the iodine-containing contrast media functions as a new excipient for taxanes. As noted above, taxanes are hydrophobic cytostatic agents. Delivering the drug in the composition having an iodine-containing contrast agent may circumvent the need for premedication to avoid the side-effects associated with currently-used excipients. Further, the use of the new fibrin matrix may allow for higher doses of paclitaxel or other taxane- containing cytostatic agents to be administered locally at a metastatic site without the need for the polyoxyethylated castor oil excipient [0043] According to one example of the present invention, the amount of contrast agent such as iodixanol (5-[N-[3-(N-[3,5-fe>[(2,3-dihydroxypropyl)carbamoyl]-2,4,6- triiodophenyl]acetamido)-2-hydroxypropyl]acetamido]-l-JV,3-JV-fc(2,3-dihydroxypropyl)- 2,4,6-triiodobenzene-l,3-dicarboxamide) or iohexol (l-JV,3-JV-fc(2,3-dihydroxypropyl)-5-[JV- (2,3-dihydroxypropyl)acetamido]-2,4,6-trϋodobenzene-l,3-dicarboxamide) in the composition ranges from about 100 to about 800 mg/mL. More preferably, the contrast agent is added in a range from about 150 to about 600 mg/mL, or more preferably still from about 200 to about 400 mg/mL, in the final composition. In one aspect, the composition is a clotted composition.

[0044] The fibrin matrix having an iodine-containing contrast media creates a porous network of fibers. This network of fibers is believed to be denser than fibrinogen alone. The density slows the release of small molecules from the matrix and can potentially be used to slow the release of small molecules from the fibrin matrix.

[0045] It is preferred that a slow and continuous release of the drug be provided. A slower release allows for local wound healing to occur at the delivery site prior to the elution of the taxane. A release time of two weeks for the taxane would be compatible with the current radiotherapeutic standard of care. However a release profile may be tailored so that faster and slower rates can be obtained.

[0046] In one aspect, the release time of the drug is affected by the concentration of the contrast agent. As the concentration of the contrast agent increases, the density of the matrix also increases. This, in turn, reduces the diffusion rate of the taxanes.

[0047] Another factor that affects the release time of the drug is the solubility of the taxanes in different iodine-containing contrast media. A high degree of solubility is desired. Therefore, a combination of a taxane and the iodine-containing contrast media is selected to increase the solubility of the taxanes. A small amount 0.1% - 1% of Cremaphor EL can be used to initially suspend the paclitaxel.

[0048] In-vitro release kinetics may be measured using analytical methods. One example is high-performance liquid chromatography, or HPLC. High-performance liquid chromatography is a form of column chromatography. HPLC is sometimes used in biochemistry to separate components of a mixture by using a variety of chemical interactions between the substance being analyzed (the analyte) and a chromatography column. [0049] In addition to the contrast agent, the fibrin matrix may optionally include other compounds. It is desirable that such additional compounds exhibit anti-neoplastic activity. It is also desirable that they be hydrophobic in nature and soluble in the iodine-containing contrast agent. The fibrin composition can also include drugs which are known to prevent bone remodeling thereby preventing further damage to the bone. An example of such a drug can be taken from the class of drugs called bisphosphonates. Drugs in this class are: Etidronate, Clodronate, Tiludronate, Pamidronate, Neridronate, Opladronate, Alendronate, Ibardronate, Risedronate, and Zolendronate). Osteonecrosis of the jaw remains a concern with the systemic delivery of bisphosphonates. However, delivery by this method ensures targeted delivery to the affected site.

[0050] The chemotherapeutic-loaded fibrin matrix can be injected locally into bones affected by metastases to treat bone pain and to prevent a progression or growth of metastases. The cured matrix with the chemotherapeutic agent provides a first line of treatment for metastatic bone whereby it will act to reduce pain and halt further bone destruction. However, the cured fibrin matrix is not intended to act as a weight-bearing substitute.

[0051] The fibrin composition may contain the various components listed above in the form of: (a) a solution; (b) a dispersion; or (c) a solid (e.g., as a lyophilisate). Any combination thereof may also be used. The components of the composition may be pre- mixed at a facility, or may be shipped as separate components where they are then mixed at a hospital or other medical facility. In one aspect, the components are pre-mixed at a laboratory or sterile facility, and then shipped in hermetically sealed vials or syringes. In another aspect, the components may be shipped or otherwise provided in separate containers as, for example, solid components. In this instance, a solvent is also provided which is separated from the components by a separation means in a container. In this instance, a solution of the combined components can be prepared by breaking or removing the separation means.

[0052] According to one example, the fibrin composition is prepared by mixing the components of the composition and homogenizing the components. The preparation of the composition can be carried out at any suitable temperature, such as in the range from about 18 to about 37 ⁰C, for example at 25 ⁰C. [0053] In another example, a solubilized contrast agent is placed in a first syringe. A thrombin/ taxane contrast agent is added to the first syringe. Fibrinogen is added in a second syringe. The two syringes are maintained separately until the time for patient treatment. At the time of treatment, the medical provider or lab technician releases the contents of the first and second syringes, where the contents are then mixed. The contents will begin to cure in the presence of oxygen and each other. The curing process typically takes place between 1 and 10 minutes depending on the composition.

[0054] According to one embodiment of the present invention, fibrinogen is used in concentration of about 75 mg/mL. The amount of thrombin in the composition is also about 75 mg/mL, and such that it is at least about 25 IU/mL to about 50 IU/mL in the final clotted composition. The amount of iodine-containing contrast agent is about 300 mg/mL in the final clotted composition.

[0055] EXAMPLE 1

[0056] Exemplary Formulation

[0057] As shown in TABLE 1 above, 600 to 800 mg/mL (in double distilled water) iodine-containing contrast agent such as iodixanol or iohexol is used. Thrombin and calcium chloride are then added to the contrast agent resulting in a final concentration of 75 IU/mL thrombin and 40 niM CaCl₂. Paclitaxel is then added to this solution at a concentration ranging from 40 to 160 ng/mL. The solution is then homogenized, centrifuged to remove bubbles, and sterilized by filtration through a 0.22 μm filter. Fibrinogen solution (91 mg/mL, reconstituted with aprotinin solution) is then mixed with the thrombin/contrast agent solution in a 1:1 ratio. In this way, the contrast agent concentration in the gelled clot is halved to either 400 or 300 mg/mL. Alternatively, a 2 mL portion of the thrombin/contrast agent solution is transferred to a 5 mL syringe. Then, a 2 mL portion of the fibrinogen (91 mg/mL) is transferred to a separate 5 mL syringe. The two syringes containing the thrombin / contrast agent / and the fibrinogen are then connected via a Luer adapter, and their contents are then homogenized by passage through the Luer adapter.

[0058] The material remains liquid for approximately 1 minute. During this time it can be injected into a patient at the point of metastasis. Alternatively, after a few minutes the clotted composition can be delivered locally as a pre-formed gel. In one aspect, a plurality of fibrin compositions is cast in wells of a multiwell-plate where they are then allowed to gel. After gelling, the clotted fibrin compositions are removed from the wells.

[0059] After gelling, the mechanical properties of the clotted compositions may be tested. For example, the clots may be placed in compression using a universal mechanical testing system. The elasticity of the clots may also be examined using non confined uni- axial compressive testing. [0060] EXAMPLE 2

[0061] Exemplary Formulation

[0062] In this formulation, a commercially available Hemaseel APR Kit (Haemacure Corp.) is utilized. Briefly, the freeze-dried Sealer Protein concentrate (150 - 230 mg fibrinogen, from Hemaseel APR Kit) was reconstituted in 2mL fibrinolysis inhibitor solution (Hemaseel APR Kit) at 37⁰C to make a fibrin solution. Paclitaxel (LC Labs) is suspended in a 1:1 mixture of Cremophor® EL:Ethanol (Sigma-Aldrich cat. no. C5135). This solution was diluted 1:10000 in a solution of Optiprep (60 mg/mL iodixanol, Sigma-Aldrich cat. no. D1556) and calcium chloride (Sigma-Aldrich), such that the final concentration was 85.4 ng/mL paclitaxel (PTX), 0.005% Cremaphor EL, 0.005% Ethanol, 99.9% Optiprep, and 4OmM calcium chloride. Thrombin (1000 IU, from Hemaseel APR Kit) was reconstituted in 2mL of the PTX / cremaphor / ethanol / optiprep / calcium chloride solution at 37⁰C, yielding a thrombin solution. The thrombin and fibrin solutions were then combined 1:1 to constitute the final formulation. The final PTX concentration in this formulation is 42.7 ng/mL, but could range between 25 and 50 nM (21.35 ng/mL and 42.7 ng/mL). Nevertheless, persons having ordinary skill in the art will recognize that the concentration of chemotherapeutic agent may be adjusted appropriately to yield a therapeutically effective dose.

[0063] EXAMPLE 3 [0064] Degradation Study

[0065] The formulation was made as described in EXAMPLE 2. 50OuL of the 1:1 fibrin:thrombin solution was added to 11mm diameter circular plastic molds. These were allowed to dry overnight at 37⁰C with 95% humidity. The resulting dried discs were then removed from the molds and placed in 2mL PBS and incubated at 37⁰C with constant agitation for 14 days. The discs were examined visually, daily, to assess degradation. No degradation was apparent over the first 7 days. During days 7 through 14 the discs underwent a gradual reduction in size, with an approximately 50% reduction in size noted by day 14.

[0066] All references cited in this specification are herein incorporated by reference as though each reference was specifically and individually indicated to be incorporated by reference. The citation of any reference is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such reference by virtue of prior invention.

[0067] It will be understood that each of the elements described above, or two or more together may also find a useful application in other types of methods differing from the type described above. Without further analysis, the foregoing will so fully reveal the gist of the present invention that others can, by applying current knowledge, readily adapt it for various applications without omitting features that, from the standpoint of prior art, fairly constitute essential characteristics of the generic or specific aspects of this invention set forth in the appended claims. The foregoing embodiments are presented by way of example only; the scope of the present invention is to be limited only by the following claims.

Claims

CLAIMSThe invention claimed is:

1. A resorbable composition comprising:

a) fibrinogen;

b) thrombin;

c) a contrast agent; and

d) a chemotherapeutic agent.

2. The resorbable composition of claim 1, wherein said contrast agent contains iodine.

3. The resorbable composition of claim 2, wherein said contrast agent is selected from the group consisting of iodecol, iodixanol, iofratol, iogulamide, iohexol, ioxaglate, iomeprol, iopamidol, iopromide, iotrol and ioversol.

4. The resorbable composition of claim 2, wherein said contrast agent is present in an amount from about 100 mg/mL to about 800 mg/mL.

5. The resorbable composition of claim 2, wherein said contrast agent is present in an amount from about 150 mg/mL to about 600mg/mL.

6. The resorbable composition of claim 2, wherein said contrast agent is present in an amount from about 200 mg/mL to about 400 mg/mL.

7. The resorbable composition of claim 2, wherein said chemotherapeutic agent is a taxane.

8. The resorbable composition of claim 7, wherein said taxane is selected from the group consisting of paclitaxel, docetaxel, milataxel, ortataxel, and protaxel.

9. The resorbable composition of claim 7, wherein said fibrinogen is present in an amount from about 5 mg/mL to about 100 mg/mL.

10. The resorbable composition of claim 7, wherein said fibrinogen is present in an amount from about 15 mg/mL to about 75 mg/mL.

11. The resorbable composition of claim 7, wherein said fibrinogen is present in an amount from about 35 mg/mL to about 63 mg/mL.

12. The resorbable composition of claim 7, wherein said thrombin is present in an amount from about 1 IU/mL to about 1000 IU/mL.

13. The resorbable composition of claim 7, wherein said thrombin is present in an amount from about 30 IU/mL to about 300 IU/mL.

14. The resorbable composition of claim 7, wherein said thrombin is present in an amount from about 100 IU/mL to about 250 IU/mL.

15. The resorbable composition of claim 7, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in an amount from about 10 mg/mL to about 200 mg/mL.

16. The resorbable composition of claim 7, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in an amount from about 30 mg/mL to about 150 mg/mL.

17. The resorbable composition of claim 7, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in an amount from about 75 mg/mL to about 115 mg/mL.

18. A method of treating cancer in a mammal in need of such treatment, comprising administering to said mammal an effective amount of a resorbable composition comprising:

a) fibrinogen;

b) thrombin;

c) a contrast agent; and

d) a chemotherapeutic agent.

19. The method of claim 18, wherein said cancer is a bone cancer.

20. The method of claim 18, wherein said cancer is a skeletal metastasis.

21. The method of claim 20, wherein said administering comprises placing said resorbable composition into a bone affected by said skeletal metastasis.

22. The method of claim 21, wherein said contrast agent contains iodine.

23. The method of claim 21, wherein said contrast agent is selected from the group consisting of iodecol, iodixanol, iofratol, iogulamide, iohexol, ioxaglate, iomeprol, iopamidol, iopromide, iotrol and ioversol.

24. The method of claim 22, wherein said contrast agent is present in an amount from about 100 mg/mL to about 800 mg/mL.

25. The method of claim 22, wherein said contrast agent is present in an amount from about 150 mg/mL to about 600mg/mL.

26. The method of claim 22, wherein said contrast agent is present in an amount from about 200 mg/mL to about 400 mg/mL.

27. The method of claim 22, wherein said chemotherapeutic agent is a taxane.

28. The method of claim 27, wherein said taxane is selected from the group consisting of paclitaxel, docetaxel, milataxel, ortataxel, and protaxel.

29. The method of claim 27, wherein said fibrinogen is present in an amount from about 5 mg/mL to about 100 mg/mL.

30. The method of claim 27, wherein said fibrinogen is present in an amount from about 15 mg/mL to about 75 mg/mL.

31. The method of claim 27, wherein said fibrinogen is present in an amount from about 35 mg/mL to about 63 mg/mL.

32. The method of claim 27, wherein said thrombin is present in an amount from about 1 IU/mL to about 1000 IU/mL.

33. The method of claim 27, wherein said thrombin is present in an amount from about 30 IU/mL to about 300 IU/mL.

34. The method of claim 27, wherein said thrombin is present in an amount from about 100 IU/mL to about 250 IU/mL.

35. The method of claim 27, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in an amount from about 10 mg/mL to about 200 mg/mL.

36. The method of claim 27, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in an amount from about 30 mg/mL to about 150 mg/mL.

37. The method of claim 27, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in an amount from about 75 mg/mL to about 115 mg/mL.

38. A method of making a resorbable composition comprising fibrinogen, thrombin, a contrast agent, and a chemotherapeutic agent, the method comprising:

a) reconstituting said fibrinogen, to make a fibrinogen solution;

b) reconstituting said thrombin, to make a thrombin solution;

c) mixing a therapeutically appropriate amount of the chemotherapeutic agent with the contrast agent, to make a chemotherapeutic agent solution;

d) mixing the thrombin solution with the chemotherapeutic agent solution to make a working thrombin solution; and

e) mixing the working thrombin solution with the fibrinogen solution.

39. The method of claim 38, wherein said contrast agent contains iodine.

40. The method of claim 39, wherein said contrast agent is selected from the group consisting of iodecol, iodixanol, iofratol, iogulamide, iohexol, ioxaglate, iomeprol, iopamidol, iopromide, iotrol and ioversol.

41. The method of claim 39, wherein said contrast agent is present in the resorbable composition in an amount from about 100 mg/mL to about 800 mg/mL.

42. The method of claim 39, wherein said contrast agent is present in the resorbable composition in an amount from about 150 mg/mL to about 600mg/mL.

43. The method of claim 39, wherein said contrast agent is present in the resorbable composition in an amount from about 200 mg/mL to about 400 mg/mL.

44. The method of claim 39, wherein said chemotherapeutic agent is a taxane.

45. The method of claim 44, wherein said taxane is selected from the group consisting of paclitaxel, docetaxel, milataxel, ortataxel, and protaxel.

46. The method of claim 43, wherein said fibrinogen is present in the resorbable composition in an amount from about 5 mg/mL to about 100 mg/mL.

47. The method of claim 43, wherein said fibrinogen is present in the resorbable composition in an amount from about 15 mg/mL to about 75 mg/mL.

48. The method of claim 43, wherein said fibrinogen is present in the resorbable composition in an amount from about 35 mg/mL to about 63 mg/mL.

49. The method of claim 43, wherein said thrombin is present in the resorbable composition in an amount from about 1 IU/mL to about 1000 IU/mL.

50. The method of claim 43, wherein said thrombin is present in the resorbable composition in an amount from about 30 IU/mL to about 300 IU/mL.

51. The method of claim 43, wherein said thrombin is present in the resorbable composition in an amount from about 100 IU/mL to about 250 IU/mL.

52. The method of claim 43, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in the resorbable composition in an amount from about 10 mg/mL to about 200 mg/mL.

53. The method of claim 43, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in the resorbable composition in an amount from about 30 mg/mL to about 150 mg/mL.

54. The method of claim 43, wherein said fibrinogen and said thrombin combine, said combination yielding fibrin in the resorbable composition in an amount from about 75 mg/mL to about 115 mg/mL.