ES2354193T3 - USE OF EPOTILONES FOR THE TREATMENT OF CANCER. - Google Patents

Abstract

Use of epothilone B for the preparation of a pharmaceutical preparation for use in the treatment of a proliferative disease where epothilone B is used in a human dose that is calculated according to formula (I) ** (See formula) ** where N is the number of weeks between treatments and and is 6.

Description

Use of epothilones for treatment of cancer

Summary of the Invention

The present invention refers to treatment of a proliferative disease, especially of according to certain treatment regimens that use epothilone B; preferably from a gastrointestinal tumor, more preferably (1) a tumor of the colon and / or rectum (colorectal tumor), especially if it is refractory to (i.e. at least one) a representative of the taxane class of anticancer agents, in particular TAXOL® (paclitaxel in form formulated for use clinical), and / or at least one standard treatment with another agent chemotherapeutic, especially 5-fluorouracil; (2) a genitourinary tract tumor, more preferably a prostate tumor, including primary and metastatic tumors, especially if it is refractory to hormonal treatment ("hormone-refractory prostate cancer") and / or treatment with other standard chemotherapeutic agents; (3) a tumor epidermoid, more preferably an epidermoid head tumor and neck, more preferably a mouth tumor; (4) a tumor of lung, more preferably a non-cell lung tumor small, especially any of these tumors that are refractory to treatment with one or more of the other agents chemotherapeutics (especially due to resistance to multiple drugs), especially when treating with a member of the taxane class of anticancer agents, in particular TAXOL®; or (5) a breast tumor, more preferably one that is resistant to multiple drugs, especially refractory to treatment with a member of the taxane class of anticancer agents, in particular TAXOL®; which is especially related to the treatment of a multi-drug resistant lung tumor (preferably a non-small cell lung tumor), a multi-drug resistant breast tumor, or a tumor multi-drug resistant epidermoid, or in one more sense of the invention, to a treatment program for the treatment of a tumor mentioned above or (in a more sense broad of the invention) any other tumor, especially if it is refractory to one or more chemotherapeutic agents, especially multi-drug resistant and / or refractory to TAXOL®, such like melanoma, ovarian cancer, pancreas cancer, neuroblastoma, head and neck cancer or bladder cancer, or in one more sense broad, renal, cerebral or gastric cancer; by means of the administration of epothilone B; the term "treatment" also covers (i) a method of treatment for (= to treat) said disease comprising the administration of said agent cytotoxic (epothilone B, in each case preferably together with a pharmaceutically acceptable excipient) to a blood animal hot, especially if you need such treatment, in a therapeutically effective amount, in at least one treatment; (ii) the use of said cytotoxic agent, for the treatment of a proliferative disease; (iii) the use of said cytotoxic agent for the manufacture of a preparation pharmaceutical for the treatment of said proliferative disease (comprising mixing said cytotoxic agent with a pharmaceutically acceptable excipient); (iv) a preparation pharmaceutical comprising a dose of said cytotoxic agent that It is appropriate for the treatment of said proliferative disease. The invention is directed, in a preferred embodiment, to a treatment of patients (humans) or groups of patients where they have other treatments failed, especially standard treatments with other chemotherapeutic agents, especially 5-fluorouracil; or therapy with class members taxane of anticancer agents, such as TAXOL®. Too refers to an epothilone, especially epothilone B, for its use in the treatment of a proliferative disease, especially where said disease is refractory to a treatment With a standard therapy.

Background of the invention

Cancer still represents a major medical problem to solve. The initial treatment of the disease is usually surgery, irradiation treatment or a combination of both, but recurrent (metastatic) disease is common. Chemotherapeutic treatments for most cancers are not generally curative, but only slow the progression of the disease. Normally, tumors and their metastases become refractory to chemotherapy, in what is known as development of resistance to multiple drugs. In many cases, the tumors are intrinsically resistant to some classes of chemotherapeutic agents [see DeVita VT, Principles of Cancer Management: Chemotherapy. In: Cancer. Principles and Practice of Oncology. DeVita VT et al (eds.), 5th edition, Lippincott-Raven, Philadelphia, New York (1977), pp. 333-347; o Cleton, FJ, Chemotherapy: general aspects. In: Oxford Textbook of Oncology; Peckham, M., et al , Oxford University Press, Oxford, New York, Tokyo (1995), Vol. 1, pp. 445-453]. This is, for example, the case for lung tumors, especially non-small cell lung carcinoma, or also for epidermoid tumors, such as head and neck epidermoid tumors, especially of the mouth, or also for tumors of the mom. Other mechanisms by which tumors are not treatable (they are refractory to treatment) may be, for example, the presence of tubulin mutations or glutathione-mediated mechanisms.

Intestinal cancer, especially colorectal, defines a special case of medical needs in the treatment of cancer not yet satisfied. The initial treatment of the disease is usually surgery, irradiation treatment or a combination of both, but recurrent (metastatic) disease is common. First-line chemotherapeutic treatments for recurrent colorectal cancer include 5-fluorouracil. But this treatment provides, at best, the delay in disease progression since the tumors usually become refractory to the treatment. Chemotherapy of this refractory stage of the disease involves the use of other classical cytotoxic agents, but all are considered inadequate [see Cohen et al ., Cancer of the colon. In: Cancer. Principles and Practice of Oncology; Delta et al . (eds.), 5th edition, Lippincott Raven. Philadelphia, New York 1997, pp. 1144-1197; or Rowinsky, Ann. Rev. Med. 48, 353-74 (1997)]. Also for genitourinary tract cancer, especially prostate cancer, another medical need not yet resolved, the initial treatment is as mentioned above for colorectal cancer, which shows similar problems. First-line chemotherapeutic treatment for recurrent prostate cancer includes antiandrogens, and recurrence is often androgen dependent. But this treatment only delays the progression of the disease since the tumors almost always become refractory to antiandrogens in a period within 6 months to 2 years (hormone refractory prostate tumors). Chemotherapy of this stage refractory to disease antiandrogens involves the use of mitoxantrone or other classic antineoplastic cytotoxic agents, but all are considered inappropriate [see Oesterling et al ., Cancer of the prostate. In: Cancer. Principles and Practice of Oncology. DeVita, VT, et al . (eds.), 5th edition. Lippincott-Raven, Philadelphia, New York 1997, pp 1322-86; Stemberg, Cancers of the genitourinary tract. In: Cavalli et al . (eds.), Textbook of Medical Oncology; or Roth, BJ, Semin. Oncol. 23 (6 Suppl. 14), 49-55 (1996)].

Among cytotoxic agents for tumor treatment, TAXOL® (paclitaxel), a microtubule stabilizing agent, has become a very important compound with extraordinary economic success [see Rowinsky EK, The development and clinical utility of the taxano class of antimicrotubule chemotherapy agents; Ann. Rev. Med. 48, 353-374 (1997)]. D. Su et al ., Angew. Chem. Int. Ed. Engl., 36 (19) 2093-2096 (1997) - exposes the structure-activity relationships of epothilones and the first in vivo comparison with Paclitaxel .

However, TAXOL® has a series of disadvantages Especially, its extremely low solubility in Water represents a serious problem. It has become necessary administer TAXOL® in a formulation with CremophorEL® ( polyoxyethylated beaver; BASF, Ludwigshafen, Germany) that has severe side effects, which causes, among other things, allergic reactions that in one case were even reported to have caused The death of a patient. More severely, it is known that certain Types of tumors are refractory to treatment with TAXOL® even when the drug is given as first-line therapy, or tumors develop resistance to TAXOL® after multiple exposure cycles

Although the taxane class of antimicrotubule anticancer agents has been hailed as "perhaps the most important contribution to the chemotherapeutic armamentarium against cancer in recent decades" [see Rowinsky EK, Ann. Rev. Med. 48, 353-374 (1997)] and despite the commercial success of TAXOL®, there are still limitations to the effectiveness of TAXOL ®. Treatment with TAXOL® is associated with a number of important side effects and some of the main categories of solid tumors, namely colon and prostate, are poorly receptive to this compound (see Rowinsky EK, loc. Cit.). Specifically, as a single agent, TAXOL® has been considered as poorly clinically active in colorectal, renal, prostate, pancreatic, gastric and brain cancers [see Rowinsky EK, loc. cit .; Bitton, RJ, et al ., Drug Saf. 12, 196-208 (1995); or Arbuck, SG, et al ., J. Natl. Cancer Inst. Monogr. 15, 11-24 (1993)]. For example, the effectiveness of TAXOL® can be seriously limited by the acquired drug resistance mechanisms that occur through different mechanisms, such as the overexpression of phosphoglycoproteins that function as drug flow pumps.

Therefore, there is an urgent need to find appropriate compounds and dosage regimens with these compounds to expand the armamentarium of the treatment of cancer, especially in most cases where the treatment with taxanes and other anticancer compounds is not associated with long-term survival.

Epothilones, especially epothilones A and B, represent a new class of microtubule stabilizing cytotoxic agents (see Gerth, K. et al ., J. Antibiot. 49, 560-3 (1996); or Hoefle et al ., DE 41 38 042), for example with the formulas:

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one

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where R is hydrogen (epothilone A) or methyl (epothilone B).

These compounds have the following advantages:

(i)

show better water solubility than TAXOL, and are therefore more appropriate for the formulation: Y

(ii)

It has been reported, in cell culture experiments, that they are also active against the proliferation of cells that, due to the activity of the P-glycoprotein effect pump that makes them resistant to multiple drugs, show resistance to treatment with other chemotherapeutic agents , for example TAXOL® [see Bollag, DM, et al ., "Epothilones, a new class of microtubule-stabilizing agents with a Taxol-like mechanism of action", Cancer Research 55, 2325-33 (1995): and Bollag DM , Exp. Opin. Invest. Drugs 6, 867-73 (1997)]; Y

(iii)

Despite apparently sharing the same site, or a sterically close binding site on the microtubule, the epothilones have been shown to be active against a TAXOL® resistant ovarian carcinoma cell line with an altered β-tubulin [see Kowalski , RJ, et al ., J. Biol. Chem. 272 (4), 2534-2541 (1997)].

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On the other hand, they are highly toxic and therefore their usefulness in the treatment of cancer in vivo was considered practically impossible [see, for example, PNAS 95, 9642-7 (1998)]. Therefore, the present invention shows in an unexpected way that it is indeed possible to find the dose regimens that allow, on the one hand, to treat tumors with epothilones, especially epothilone B; and on the other hand it allows the treatment of certain groups of patients that do not respond to other types of treatments, whether due to resistance to multiple drugs, such as taxane, for example TAXOL®, refractoriness due to resistance to multiple drugs, and / or any other mechanism

The present invention aims to present for the first time in vivo regimens for a useful treatment with epothilone B, which allows the treatment of a tumor disease, for example a melanoma, ovarian cancer, pancreas cancer, neuroblastoma, head cancer and neck, bladder cancer, renal, cerebral, gastric or preferably colorectal cancer, prostate cancer, breast cancer, lung cancer (especially non-small cell lung cancer) or epidermoid cancer, for example epidermoid head cancer and neck, especially epidermoid mouth cancer.

Although the general treatment program allows the treatment of different types of tumors already in the first line treatment, the present invention makes reference, preferably, to the treatment of tumors that can be expect them to be, or have proven to be, refractory to treatment with other chemotherapeutic agents, for example the standard treatment with one or more chemotherapeutic agents, especially with 5-fluorouracil and / or taxane, for example the treatment with TAXOL®.

Surprisingly, it has now been found that even the proliferation of tumor cells and tumors that are refractory to standard treatment with other chemotherapeutic agents, for example 5-fluorouracil; and / or to the treatment with a member of the compounds of the taxane class, especially TAXOL®, especially of a colorectal tumor, especially one that is also refractory to the standard treatment, for example with 5-fluorouracil; or of a lung tumor, especially a non-small cell lung cancer; an epidermoid tumor, more preferably epidermoid head and neck, such as the mouth tumor; or a breast tumor; and / or metastasis can be weakened or stopped and that even regression or disappearance of the
tumor.

Detailed description of the preferred aspects of the invention

The present invention preferably addresses the following theme as part of the invention:

Whenever it is mentioned within all this specification "the treatment of a proliferative disease" or of a tumor, cancer or the like, it is understood

to)

the use of epothilone B, for the treatment of a proliferative disease, or epothilone B, for use in the treatment of said disease (especially in a human); in a dose that allows the treatment of said disease (= one therapeutically effective amount), preferably in a dose (quantity) that was specified as preferred before and at continued in this document;

b)

the use of epothilone B, for the elaboration of a pharmaceutical preparation for the treatment of a proliferative disease; in a dose that allows the treatment of said disease (= a therapeutically effective amount), preferably in a dose (amount) that was specified as preferred before and then in this document; the which means that the respective tumor disease can be located under the above a) to b) instead of "disease proliferative ", according to the patentable subject: preferably, any treatment under a) to b) refers to a treatment of humans.

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This application describes the treatment of a proliferative disease that is refractory to treatment with one or more chemotherapeutic agents, where epothilone B, is administered to a hot-blooded animal, especially a human, preferably a human who needs such treatment, especially in a therapeutically effective amount.

The present invention refers to an in vivo regimen for the treatment of a proliferative disease, especially a cancer that is refractory to treatment with one or more chemotherapeutic agents, especially of the taxane class, such as TAXOL®, and / or 5-fluorouracil. , where epothilone B, is administered in a dose that is between about 1 to 100%, preferably between about 25 to 100%, of the maximum tolerated dose (MTD) (in a single administration) to a warm-blooded animal, especially a human: and one or more (preferably two to seven) additional doses, preferably each within the aforementioned dose range, are administered in one or, preferably, more than one cycle or cycles of additional treatment, especially with an interval between treatment cycles of one week or more than one week after the previous treatment, more preferably after about one to 6 s emanates, more preferably about one to three weeks after the previous treatment, respectively. Generally, this treatment regimen is preferred where a high dose is administered in two or more treatment cycles with periods of time between one to six, preferably one to three weeks between administrations of the most frequent treatments with lower doses. , especially since it should reduce the frequency and duration of hospitalization and since it shows superior antitumor effects and less toxicity than more frequent treatments, and greater antitumor efficacy than less frequent treatment.

Preferably, for epothilone B the dose which is used in humans is calculated according to the formula (I)

2

where N (an integer (such as 1, 2 or 3) or fraction (such as 1.5 or 2.3) whenever it is mentioned with before and then in this document)) is the number of weeks between treatments (preferably a number in the range from 1 to 6 (corresponding to a range of about 1 to about 6 weeks), especially 1 to 3 (corresponding to a preferred range of 1 to 3 weeks) and y is 6 or preferably 5, more preferably Four.

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More preferably, the treatment dose is Calculates according to formula II

3

even more preferably of according to the formula III,

4

or more preferably agree with the formula IV

5

where, in each formula from II to IV, N has the meaning given under formula I. For doses calculated according to any of the formulas from I to IV, the following condition must be met: The dose, even if It is calculated higher, it should not exceed about 18 mg / m2 for a single administration.

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Preferably, for the weekly treatment the dose is between 0.1 and 6, preferably between 0.1 and 5 mg / m2, more preferably between 0.1 and 3 mg / m2, even more preferably between 0.1 and 1.7 mg / m2, more preferably between 0.3 and 1 mg / m2; for the three week treatment (treatment every three weeks or every third week) the dose is between 0.3 and 18 mg / m2, preferably between 0.3 and 15 mg / m2, more preferably between 0.3 and 12 mg / m2, even more preferably between 0.3 and 7.5 mg / m2, even more preferably between 0.3 and 5 mg / m2, more preferably between 1.0 and 3.0 mg / m2. This dose is preferably administered at human by intravenous (iv) administration for 2 to 180 minutes, preferably 2 to 120 minutes, more preferably for 5 to 30 minutes, more preferably for 10 to 30 minutes, for example for about 30 minutes.

Preferably, especially in the case of weekly treatment, periods of more than one week, more preferably two to ten weeks, more preferably three to six weeks after the previous treatment may be necessary after for example 3, 4, 6, 8, or more treatment cycles, according to the patient's condition, to allow sufficient recovery from previous treatment.

This application describes the in vivo regimen for the treatment of a proliferative disease, especially one that is refractory to treatment with one or more chemotherapeutic agents, where epothilone B, is administered weekly to a warm-blooded animal, especially a human, in a dose that is below 80%, more preferably below 50% of the maximum tolerated dose (MTD).

This application describes the in vivo treatment of a proliferative disease that is refractory to treatment with one or more chemotherapeutic agents, especially 5-fluorouracil or a microtubule stabilizing agent of the taxane class, especially TAXOL®, for example a multi-resistant tumor. drugs, where epothilone B, is administered to a warm-blooded animal, especially a human.

This application describes the in vivo treatment of a proliferative disease, especially one that is refractory to treatment with one or more chemotherapeutic agents, by the combined administration (a) of epothilone B, in combination with (b) another antitumor chemotherapeutic, preferably being the intended treatment so that component (a) and (b) are administered to a warm-blooded animal, especially a human (especially in need of such treatment), in combination in an amount that is altogether therapeutically effective against a proliferative disease that it can be preferably treated by administration of epothilone B; such administration being performed to a human having a tumor that is refractory to other chemotherapeutic treatments, for example to treatment especially with 5-fluorouracil or especially with a member of the taxane class of anti-cancer agents, such as TAXOL®.

This application describes a preparation combined comprising components (a) and (b) as defined in the previous paragraph.

This application describes a product that comprises component (a) and component (b) as defined previously in the preceding paragraph before, in the presence or absence of one or more pharmaceutically acceptable excipients, as a combined preparation for administration simultaneously or chronologically progressive to a blood animal hot, especially a human, within a period of time which is small enough for active compounds both component (a) and component (b) increase mutually antiproliferative activity (especially against proliferating cells) in said warm-blooded animal, to The treatment of a proliferative disease.

The general terms used previously and then in this document they preferably have the following meanings, unless otherwise defined:

An illness Proliferative is primarily a tumor disease (or cancer) (and / or any metastasis), wherever the tumor or metastasis be located), more especially a tumor selected from the group comprising breast cancer, genitourinary cancer, cancer lung, gastrointestinal cancer, epidermoid cancer, melanoma, ovarian cancer, pancreatic cancer, neuroblastoma, cancer head and neck (this term, whenever used, means a head and / or neck cancer, that is, not only a head and neck cancer, but also head or neck) or cancer of bladder, or in a broader sense kidney, brain or gastric; more preferably (i) a tumor selected from a breast tumor; an epidermoid tumor, especially an epidermoid tumor of head and neck, preferably a mouth tumor; and a tumor of lung, especially a non-small cell lung tumor, or from among a gastrointestinal tumor, especially a tumor colorectal; and a genitourinary tumor, especially a tumor of prostate (especially a prostate tumor refractory to hormones); or (ii) (more preferably) a proliferative disease which is refractory to treatment with other agents chemotherapeutics, especially a corresponding tumor (and / or any metastasis), more especially a tumor selected from among the group that includes tumors that are refractory to standard treatment with other agent (s) chemotherapeutic (s), especially with 5-fluorouracil and / or (preferably) an agent microtubule stabilizer of the taxane class, plus especially TAXOL®, even more preferably a selected tumor from among a gastrointestinal cancer, for example colorectal cancer (especially refractory to standard treatment, for example with 5-fluorouracil, and / or TAXOL®); and cancer genitourinary, for example prostate tumors (and / or a metastasis of them, especially a metastasis thereof); plus preferably a gastrointestinal tumor, especially a cancer colorectal; or (iii) a tumor that is refractory to treatment with other chemotherapeutic agents due to multiple resistance drugs, especially refractory to a member of the class of taxane of microtubule stabilizing agents, preferably TAXOL®, more especially to multiple drugs, especially TAXOL®, resistant lung tumor (especially a non-small cell lung tumor), a resistant breast tumor to multiple drugs, or a multiple resistant epidermoid tumor drugs, preferably head and neck epidermoid, more preferably from the mouth.

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In a broader sense of the invention, a proliferative disease can also be selected from among hyperproliferative diseases such as hyperplasias, fibrosis (especially the lungs, but also other types of fibrosis, such as renal fibrosis), angiogenesis, psoriasis, atherosclerosis and smooth muscle proliferation in the vessels blood, such as stenosis or restenosis after angioplasty

Where mentioned earlier and later in this document a tumor, a tumor disease, a carcinoma or a cancer, it is assumed alternatively or in addition metastasis in the original organ or tissue and / or any other location, regardless of tumor location and / or metastasis.

The word "refractory" means that the respective proliferative disease (especially a tumor and / or any metastasis of it), before treatment with a (is say, at least one) chemotherapeutic agent that is not a epothilone, shows no or only a weak response antiproliferative (no or only a weak inhibition of tumor growth) after treatment with said agent, it is that is, a tumor that cannot be treated at all or only with Unsatisfactory results with other chemotherapeutic agents (preferably standard) (preferably as defined previously, especially 5-fluorouracil (especially in the case of colorectal cancer, such as cancer of colon), antiandrogens, such as letrozole (especially in the case of breast cancer); or especially a class member of the taxane of chemotherapeutic agents, for example TAXOTERE® or TAXOL®, in a warm-blooded animal, especially a human; for example, tumor growth does not stop, it is only delayed slightly or no regression is discovered. In the present invention, where the treatment of refractory tumors is mentioned and similar, it should be understood that it covers not only a tumor or tumors where one or more chemotherapeutic agents have already failed during the treatment of a patient, but also a tumor or tumors that proves that they are refractory by other means, for example Biopsy and culture in the presence of chemotherapeutic agents.

Where a term is used as "refractory before TAXOL® "earlier and later in this document, This term is intended to also mean paclitaxel, the active substance of TAXOL® in addition to the finished product. "Refractory to hormonal treatment" or "refractory to hormones ", in the case of a genitourinary tract tumor, especially a prostate tumor, means refractory to treatment with an antiandrogen.

TAXOL® preferably means the product final comprising paclitaxel, but, in a broader sense, It is also understood to cover the same paclitaxel of any other formulation with one or more excipients.

Preferably, the term refractory means that a tumor growth reduction of less than 50% (ie a T / C% value equal to or greater than 50%) is obtained with a standard dose when compared to a control without chemotherapeutic agent, for example by measurements in vivo or in vitro .

A multiple resistant tumor disease drugs is one where resistance to one or more is found chemotherapeutic agents, including those in the class of taxane, especially TAXOL®, or anthracycline class, especially ADRIAMYCIN®. The basis for this resistance is the export through an energy dependent pump (especially ATP) located on the surface of the cells of the respective tumor, especially from the family of the P-glycoprotein, especially the own P-glycoprotein (P-gp). In the present invention, alternatively or additionally other mechanisms they can make a tumor refractory to treatment with chemotherapeutic agents other than an epothilone. For example, alterations of the drug target (especially microtubules in the present case), changes in metabolism intracellular that could cause the compound to become inactive, or changes in cell physiology that could facilitate that the mechanism of the action of the drug is avoided or nullified can lead to such resistance.

The term "other agents chemotherapeutic agents "or" standard chemotherapeutic agent ", especially indicates any chemotherapeutic agent that is not an epothilone; preferably one as defined in the introduction, especially 5-fluorouracil (especially in the case of colorectal cancer, such as cancer of colon), an antiandrogen or a mitoxantrone (especially in the case of prostate cancer), or an antiestrogen, such as letrozole (especially in the case of breast cancer); especially the term refers to 5-fluorouracil or (more preferably) to members of the agent taxane class microtubule stabilizers, such as preferably Taxotere or more preferably TAXOL. "Standard treatment with others chemotherapeutic agents "," treatment with other agents chemotherapeutics "or" standard chemotherapy "make reference to treatment with at least one of said "other" or "standard therapy".

The term epothilone refers to the epothilone B.

The administration in all the cases mentioned before and later it can be done orally, in view of an increasingly better bioavailability, more preferably it is done parenterally, especially intravenous mode, for example by infusion or injection. Where subsequently "infusion" is used, preferably means intravenous infusion, which is the most preferred mode of administration.

Subsequently, the data for adults are The basis for the illustration. However, it is clear that the The present invention also refers to the treatment of proliferative diseases in pediatrics. The doses must be corrected according to standard methods and age, condition and others patient characteristics

The Maximum Tolerated Dose (BAT) is determined by according to standard procedures; preferably in warm-blooded animals BAT in the case of oral administration or intravenous is determined as the dose of a single administration, where death does not take place and a loss of body weight of less than 40, preferably less than 25, percent (%) in the treated warm-blooded animal individual  (This term here mainly refers to an animal; for humans see below).

BAT may vary according to patient population, which can be defined by the type of tumor, age range, sex, stage of the tumor and the like. While in animals, the most preferred way of determining BAT may be analogous to that shown in the Examples presented below, in humans BAT can generally be determined by starting with a single administration of a very low dose, for example 1 / 10 of the LD10 (i.e. the dose that is lethal for 10% of the animals) in the most sensitive animal species in which the toxicological studies have been conducted, for example for epothilones (especially epothilone B) in the range between 0.1 and 25 mg / m2, especially for epothilone B in the range between 0.1 and 2.5 mg / m2, more especially in the range of 0.1 and 0.33 mg / m2. The dose increase for the next dose level is 100%, unless a grade 2 toxicity is seen according to the Revised Common Toxicity Criteria of the United States National Cancer Institute, in which case the dose increase will be 67% The dose increase for subsequent dose levels is in the range between 25% and 67%. For example, three patients are usually treated with a dose level and the presence of acute toxicity is observed for one treatment cycle before any other patient is admitted. If none of the three patients experience DLT (dose-limiting toxicity), then the new cohort of three patients is treated with the next higher dose. If two or more of the three patients experience DLT, then three other patients are treated with the next lower dose unless six patients have already been treated with that dose. If one of three patients treated with one dose experiences DLT, then three other patients with the same level are treated. If the incidence of DLT among those patients is one in six, then the next cohort is treated with the next higher dose. In general, if two or more than six patients treated with a dose level experience DLT, then MTD is considered to have been exceeded, and three other patients are treated with the next lower dose as described above. BAT is defined as the highest dose studied for which the incidence of DLT was less than 33%. Usually the dose increase for subsequent cycles in the same patient, that is, the increase in intra-patient dose is not allowed. Alternatively, the dose steps can be defined by a modified Fibonacci series in which the dose increases for successive levels beyond the starting dose are 100%, 67%, 50% and 40%, followed by 33% for all subsequent levels. Finally, BAT can be found by methods described in Simon, R., et al ., J. Nat. Cancer Inst. 89 (15), 1997,
p. 1138-1147.

DLT generally includes (but is not limited to a) any death related to a drug and most of the Grade 3 and 4 toxicities related to a drug, including febrile neutropenia (see also Common Toxicity Criteria Revised from the National Cancer Institute of the United States). Watch Especially the examples. For a human, the doses of Preferred treatments are defined by formula I, plus preferably formula II, more preferably formula III mentioned above (with the proviso that no dose is greater than 18 mg / m2). Preferably, for the treatment Weekly the dose is between about 0.1 and 6, preferably between about 0.1 and 5 mg / m2, more preferably between about 0.1 and 3 mg / m2, even more preferably between about 0.1 and 1.7 mg / m2, more preferably between about 0.3 and 1 mg / m2; for the treatment of three weeks (treatment every third week) the dose is between about 0.3 and 18 mg / m2, preferably between about 0.3 and 15 mg / m2, more preferably between about 0.3 and 12 mg / m2, even more preferably between about 0.3 and 7.5 mg / m2, even more preferably between about 0.3 and 5 mg / m2, more preferably between about 1.0 and 3.0 mg / m2. This dose is administered at preferably human by intravenous administration (iv) for 2 to 180 minutes, preferably 2 to 120 minutes, preferably for about 5 to 30 minutes, more preferably for about 10 to 30 minutes, for example for about 30 minutes.

From the use of animal data, Applicable doses in humans (adults) can be calculated approximately as follows:

A dose of 1 mg / kg in a mouse corresponds to a dose of 3 mg / m2 in a human.

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It is understood by sufficient recovery of the previous treatment, in warm-blooded animals especially the recovery of the body weight of the individual treated at the level of start found before the first administration, preferably at least 95% of said weight. In a human, the recovery of each previous dose administration is defined preferably as recovery from any toxicity  grade of 3 or 4, including for example the achievement of a count of platelets of at least 100,000 / mm3 and a count of neutrophils of at least 1,500 cells / mm 3 in blood complete.

The treatment can be repeated if it is not achieved a response after a first treatment, until it find a tumor progression or until other reasons (for example the patient's condition) require the interruption of the treatment. In a human, treatment with about 25 to about 100% of the BAT is preferably repeated every 1 to ten, especially 2 to ten weeks; preferably every 1 to 10 weeks, or every 3 to 6 weeks, until the disease progression, unacceptable toxicity, 1 or preferably 2 cycles after termination of a response complete, or the revocation of the patient's consent by any reason

Preferably, in the case of treatment Weekly of a human with epothilone, the dose is in the range of between about 5 and 60%, preferably between approximately 10 to 60%, for example between approximately 5 and 35% of BAT, especially in the range between approximately 30 and 35% of BAT. Preferably, for the epothilone B the dose is in the range of about 5 and 60%, more preferably between about 10 and 60%, especially in the range of about 10 to 45%, more especially in the range of about 30 to 45% of the three weekly BAT.

More preferably, the treatment after the third to the eighth, especially after  from the third to the fifth weekly administration followed by a rest period from two to five, for example two weeks before that treatment be resumed, by single administration or either again weekly or two per week. Especially in the case of weekly epothilone B treatment stops after the third to the eighth administration followed by a period of break two to four, for example two weeks before it Resume treatment through weekly administration.

The administration of component (a), that is epothilones B, preferably takes place as described with previously, especially by using one of the special treatment regimens mentioned above.

The administration of component (b) preferably it takes place according to the programs of treatment that are known to the person with knowledge in the art.

In a preferred embodiment, component (b) it is administered before component (a), preferably in a treatment comprising one or more administrations of the component (b) before beginning treatment with component (a), preferably such treatment with component (b) ends by at least two, preferably 5 to 10, for example approximately 5 days before treatment with component (a) which is administered one or more times later, preferably from one to five, especially one to two times.

In a more preferred embodiment, the component (a) is administered every 3 weeks before component (b), preferably in a treatment comprising an administration of component (a) before the start of treatment with the component (b), more preferably such treatment with the component (a) ends immediately before treatment with the component (b) that is administered later.

In a second more preferred embodiment, the Component (a) is administered based on a weekly program. He component (b), on the other hand, is administered every 3 weeks, where each administration is done immediately after the completion of each third administration of component (a).

In a third more preferred embodiment, the component (a) is administered based on a weekly program before component (b), preferably in a treatment comprising a administration of component (a) before the start of treatment with component (b), more preferably such treatment with the component (a) ends immediately before treatment with the component (b) that is administered later.

The term "other chemotherapeutic agent" It is used to refer especially to any agent chemotherapeutic which is used or can be used in the treatment of tumor diseases, such as agents chemotherapeutics derived from the following categories:

(TO)

Alkylating agents, preferably agents cross-linking chemotherapeutics, preferably agents bis-alkylating agents,

(B)

antitumor antibiotics, preferably the doxorubicin (ADRIAMYCIN®, RUBEX®);

(C)

antimetabolites;

(D)

plant alkaloids;

(AND)

hormonal agents and antagonists,

(F)

biological response modifiers, preferably lymphokines or interferons

(G)

protein tyrosine kinase inhibitors and / or serine / threonine kinases;

(H)

antisense oligonucleotides or derivatives of oligonucleotides; or

(I)

various agents or agents with another mechanism of action or an unknown one.

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It is preferably understood by the term "therapeutically effective jointly against a disease  proliferative that can be treated by administering epothilone B, ", a proliferative disease as mentioned previously, especially a tumor disease, also expressing the response preferably in a reduced proliferation, for example the reduced growth of tumor or even (more preferably) tumor regression or (more preferably) the disappearance of the tumor ("response complete ").

Preferably, the term "amount that is therapeutically effective jointly against a disease proliferative that can be treated by administering epothilone B "means any amount of the components (a) and (b) of the combinations that, in the combination, are reducing the proliferation of the cells responsible for any of the mentioned proliferative diseases, especially cells tumor (including metastatic) (especially the reduced tumor growth) or, preferably, even causing regression, more preferably even disappearance partial or complete, of said cells (especially regression tumor, preferably the complete answer means the disappearance of the tumor (s). This term not only includes the combinations of any component (a) and (b) where (a) and (b) they are dosed in such a way that they are already antiproliferatively effective no combination, but also doses of any component that just wouldn't show any effect or just a marginal effect but which in combination leads to clearly antiproliferative effects, that is to say a reduced proliferation or preferably even at a regression of proliferating cells or even the cure of proliferative disease Also, here the term "combination" is not only used to describe combinations fixed components, but also any combination of components (a) and (b) for simultaneous use or chronologically progressive within a period of time that is small enough for the active compounds both of the component (a) and component (b) mutually increase the antiproliferative activity, for example in a patient.

It is understood by the term "preparation combined comprising component (a) and (b) "any combination, this being a set of parts or a combination unique, of component (a) and (b) in the form of a product pharmacist, which is preferably where a pharmaceutically acceptable excipient. For the excipients preferred, see below under "Preparations pharmaceuticals ".

It is preferably understood by the term "a product comprising component (a) and component (b) ", a product comprising (a) at least epothilone B and (b) so minus one of the other chemotherapeutic agents in the presence or absence of one or more pharmaceutically excipient materials acceptable, as a combined preparation, for use simultaneously or chronologically progressive, preferably within a period of time that is small enough for the active components of both component (a) and component (b) mutually intensify antiproliferative activity against proliferative cells, especially in a patient, for the treatment of a proliferative disease that responds to such active compounds '', especially a "set of parts" in the sense that the effective components (a) and (b) of the combination  they can be dosed independently or by using different fixed combinations with distinguished amounts of any of the components (a) and (b) at different points Temporary The "parts" of the set of parts can be then administer simultaneously or chronologically progressive, that is to say at different time points and with the same or different time intervals for any part of the set of parts, preferably on condition that the time intervals so that the effect on the disease proliferative in the combined use of the parts is greater that the effect that would be obtained by using only one of any of the components (a) and (b) separately or by the use of both in a way in which the compounds act independently (for example with sufficiently long periods to avoid the effects of each of the components on the others), that is, there is a stronger inhibition of proliferation or, preferably, a stronger regression or even  the cure of proliferative disease that when the same dose of only one of the components (a) and (b) is administered separately with the same dose or after sufficient time intervals long so that the reciprocal effects of the components (a) and (b). That is what is meant by the term "mutually intensify antiproliferative activity against proliferating cells, especially in a patient "; preferably a reciprocal intensification of the effect is understood of the components (a) and (b), especially a synergy and / or the obtaining a regression of proliferating cells, up to e including its complete destruction, and especially a strong synergy between components (a) and (b).

It is understood by the term "cells proliferating ", especially pathologically or abnormally cells proliferators, such as a tumor and / or tumor cells metastatic, especially tumors as defined with prior to this document as preferred.

The combinations shown are preferred intensified antiproliferative activity compared to single components alone, especially the combinations that show synergy (synergistic combinations) or combinations that lead to a regression of proliferating tissues and / or the cure of Proliferative diseases

The term "synergy" refers to an effect that is stronger than additive, that is, a stronger effect resulting from the combination of any component (a) with any component (b) of what could be achieved by the factor of Decrease in proliferation obtained from the mere multiplication of the proliferation decrease factor for any component (a) separately or any component (b) separately compared to an untreated control when each (a) and (b) as such, either alone or in combination, they are administered with the same dose as in the single treatment without combination (which does not mean that the dose of (a) must be identical to that of (b), although this could also be the case: As a theoretical example for illustrative purposes only, if a component (a) only gives a tumor cell growth that is decreased by a factor of 2 compared to a control without any treatment and a comp Onente (b) only gives a decrease in growth by a factor of 1.5, so an additive effect would be one where there would be a tripled decrease in growth (multiplication of 2 with 1.5), through the combined use of the component ( a) and component (b). A synergistic effect would be present for example if a decrease greater than triple the proliferation is discovered. The presence of synergy can be shown by this method of fractional products [Webb, in: "Enzymes and Metabolic Inhibitors", Vol. 1, 66-73 and 488-512, Academic Press, New York] or alternatively by the method of isobolograms [see references in: Berenbaum Pharmacol. Rev. 41, 99-141 (1984)], and / or the combination index calculation method (Cl) [Chou et al ., Trends Pharmacol. Sci. 4, 450-454 (1983); or Chou et al ., New Avenues in Developmental Cancer Chemotherapy; Bristol-Myers Symposium Series, KR Portuguese and IA Connors (eds.), 37-64, New York, Academic Press (1987)].

The following is explained in the definition of pharmaceutical preparations the term "excipient materials pharmaceutically acceptable. "

The component (b) (other (s) chemotherapeutic agent (s) may also be present in the form of salts whenever mentioned above or subsequently, as long as they are in the respective molecule present the groups that form salts.

The termination of the treatment takes place preferably when one of the following occurs: Progression of the disease, for example according to the response criteria of the Southwest Oncology Group (SWOG); unacceptable toxicity (for example in the patient, the researcher, or both); treatment of 2 cycles beyond the termination of a complete response, for example according to the Southwest response criteria Oncology Group (SWOG); or the revocation of the consent of the patient.

The salts of the components are especially acid addition salts, salts with bases or, when they are various salt forming groups are present, optionally also mixed salts or internal salts. The salts are especially the salts pharmaceutically acceptable, for example substantially non-salts Toxic

Such salts are formed, for example, from chemotherapeutic agents that have an acid group, for example a carboxyl, phosphodiester or phosphorothioate group, and are, by example, its salts with suitable bases, such as salts non-toxic metals derived from metals of groups Ia, Ib, Iia and Iib of the Periodic Table of the Elements, especially suitable alkali metal salts, for example salts of lithium, sodium, or potassium or ammonium salts, also those salts which are formed with organic aminos, such as mono-, di- or tri-alkylamines substituted by hydroxide or not substituted, especially mono-, di- or lower tri-alkylamines, or with compounds of quaternary ammonium, for example N-methyl-N-ethylamine, diethylamine, triethylamine, mono-, bis- or tris- (2-hydroxy-lower alkyl) amines, such as mono-, bis- or tris- (2-hydroxyethyl) amine, 2-hydroxytert-butylamine or tris (hydroxymethyl) methylamine, N, N-di-lower alkyl-N- (hydroxy-lower alkyl) -amines, such as N, N-dimethyl-N- (2-hydroxyethyl) -amine or tri- (2-hydroxyethyl) -amine, or N-methyl-D-glucamine, or quaternary ammonium salts, such as salts of tetrabutylammonium The chemotherapeutic agent that has a group basic, for example an amino or imino group, can form salts of addition of acids, for example with inorganic acids, for example a hydrohalic acid, sulfuric acid or phosphoric acid, or with organic, symphonic, sulfo or phospho carboxylic acids or N-substituted sulfamic acids such as, by example, acetic acid, propionic acid, glycolic acid, acid succinic, maleic acid, hydroximaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, gluconic acid, citric acid, or benzoic acid, also with amino acids, by example, α-amino acids, and also with acid methanesulfonic acid, ethanesulfonic acid, acid 2-hydroxyethanesulfonic acid ethane-1,2-disulfonic acid benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene-2-sulfonic acid, acid N-cyclohexylsulfamic (with the formation of cyclamates) or with other acidic organic compounds, such as ascorbic acid Compounds that have acidic groups and Basic can also form internal salts. If more than one group salt former is present, it is possible that mixed salts be present too.

When the numerical terms are used in the present, before and then, are intended include the numbers that represent the upper limits and lower. For example, "between 1 and 3" represents a range "from 1 inclusive and up to 3 inclusive 3", and "in the range from 1 to 3 "would represent" from 1 inclusive and up to 3 inclusive 3 ". The same is valid if instead of numbers (for example 3) words denoting numbers are used (for example "three").

Where "understanding" is used, this can preferably be replaced by "consisting essentially in ", more preferably by" consisting of ".

Where "approximately" is used with reference to a number, this preferably means the number ± 15%, more preferably the number plus 5%, more preferably the same number without "approximately". For example, "approximately 100" would mean "from 85 inclusive up to 115 inclusive ". Where" approximately "is used with reference to numerical ranges, for example "from about 1 to about 3 ", or" between about one and approximately three ", the definition of" approximately " with reference to a number in the last sentence applies preferably to each number that defines the beginning and end of a range separately. Preferably, where it is used "approximately" with reference to any numerical value, the "approximately" can be deleted.

"Weekly" refers to "about once a week" (which means that does more than one treatment with an interval of approximately one week between treatments), "approximately" in this case preferably means ± 1 day (that is, "every 6 to 8 days "); more preferably," weekly "refers to "once every 7 days".

"Every 3 weeks" or "every three weeks" refers to "approximately once every three weeks" (which means that more than one treatment is done with a interval of approximately three weeks between treatments), the approximately means in this case preferably ± 3 days (that is, every 18 to 24 days); more preferably, "weeks" refers to "once every 21 days" (= every third week).

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In the following preferred embodiments of the invention, the general definitions can be replaced by more specific definitions provided previously or with later in this document, as appropriate.

(1) The present invention relates especially to the treatment of a proliferative disease, especially a cancer, especially a cancer that is refractory to treatment with other chemotherapeutic agents and / or a member of the taxane class of anticancer agents, especially TAXOL®, more especially one of the preferred diseases as defined previously or subsequently, characterized in that epothilone B, is administered more than once with an interval of one to three weeks to a human in a dose that is calculated according to the formula (I)

6

where N (an integer or fraction) is the number of weeks between treatments (of approximately one to approximately three weeks), that is N is from about 1 to about 3; more preferably, the Treatment dose is calculated according to the formula II

7

even more preferably agree with the formula III,

8

or even more preferably of according to the formula IV

9

where, in each of the formulas from II to IV, N is from about 1 to about 3 (what which corresponds to the intervals of approximately 1 to approximately 3 weeks between treatments);

epothilone B, administration being made preferably

(to)

weekly in a human in a dose between about 0.1 and about 6, preferably between about 0.1 and about 5 mg / m2, more preferably between about 0.1 and about 3 mg / m2, even more preferably between about 0.1 and about 1.7 mg / m2, more preferably between about 0.3 and about 1 mg / m2; or

(b)

every three weeks in a human in a dose between about 0.3 and about 18 mg / m2, preferably between about 0.3 and about 15 mg / m2, plus preferably between about 0.3 and about 12 mg / m2, even more preferably between about 0.3 and approximately 7.5 mg / m2, even more preferably between about 0.3 and about 5 mg / m2, more preferably between about 1.0 and about 3.0 mg / m2; running the administration preferably by an intravenous infusion for 2 to 120 minutes, more preferably for about 5 to about 30 minutes, more preferably for about 10 to about 30 minutes, for example for about 30 minutes

(2) This application describes the treatment of a tumor disease that is refractory to treatment with others chemotherapeutic agents, especially selected from 5-fluorouracil and preferably an agent microtubule stabilizer of the taxane class, more especially TAXOL®, said tumor being selected from a tumor gastrointestinal, for example colorectal; a renal tumor; A tumor genitourinary, for example prostate; a pancreatic tumor; and a brain tumor (and / or any metastasis thereof), more preferably a gastrointestinal tumor, especially a cancer colorectal, more especially a gastrointestinal cancer, especially a colorectal cancer, that is, refractory to treatment with a member of the agent taxane class anticancer, especially TAXOL®, or very especially such a tumor which is refractory to a standard chemotherapy, such as a standard chemotherapeutic treatment, especially with 5-fluorouracil; or a tumor of the tract genitourinary, especially a prostate cancer, more especially a hormone-refractory prostate cancer; where the Epothilone B is given to a warm-blooded animal, especially a human.

(3) This application describes the treatment of a tumor disease, especially a lung tumor, especially a non-small cell lung carcinoma, especially such lung cancer that is refractory to treatment with a member of the taxane class of anticancer agents, especially TAXOL®; a breast tumor, especially one that is multi drug resistant; or an epidermoid tumor, preferably a head and neck epidermoid tumor, especially by mouth, especially if the latter is resistant to multiple drugs and / or resistant to treatment with a member of the taxane class of anticancer agents, in particular TAXOL®; where epothilone B is administered to a blood animal Hot, especially a human.

(4) This application describes an in vivo regimen for the treatment of a tumor disease, especially (i) of a gastrointestinal tract tumor, more especially a colon and / or rectum tumor (colorectal tumor); and / or (ii) a genitourinary tract tumor, especially a prostate tumor (preferably a hormone-refractory prostate tumor); especially where such a tumor is refractory to treatment with another chemotherapeutic agent, especially one of the taxane class, more especially TAXOL®; where epothilone B is administered once in a dose between about 20 and about 100% of the MTD, to a human; and, if required, one or more (preferably two to seven) additional doses each within the dose range mentioned above for the first dose is administered in additional treatment cycles, preferably each dose after a period of time that allows sufficient recovery of the individual under treatment of each previous dose administration, especially more than two weeks after the previous treatment, more especially two to 10 weeks, more especially three to six weeks after the previous treatment, especially three weeks after that treatment.

More preferably, under (1) to (4) the epothilone B is administered weekly to a human with a dose between about 0.1 and about 6, preferably between about 0.1 and about 5 mg / m2, plus preferably between about 0.1 and about 3 mg / m2, even more preferably between about 0.1 and about 1.7 mg / m2, more preferably between about 0.3 and about 1 mg / m2; or epothilone B It is given every three weeks (every 3 weeks) at a dose between about 0.3 and about 18 mg / m2, preferably between about 0.3 and about 15 mg / m2, plus preferably between about 0.3 and about 12 mg / m2, even more preferably between about 0.3 and approximately 7.5 mg / m2, even more preferably between about 0.3 and about 5 mg / m2, more preferably between about 1.0 and about 3.0 mg / m2. This dose is preferably administered to the human. by intravenous (iv) administration for 2 to 120 minutes, more preferably for about 5 to about 30 minutes, more preferably for about 10 to about 30 minutes, for example for about 30 minutes

More preferably, said treatment is repeated every about 1 to about 3 weeks, until the disease progression, unacceptable toxicity, 1 or preferably 2 cycles beyond the determination of a complete response, or until the revocation of the consent of the patient for any reason.

(5) This application describes an in vivo regimen for the treatment of a tumor disease, especially (i) of a gastrointestinal tract tumor, more especially a colon and / or rectum tumor (colorectal tumor); and / or (ii) a genitourinary tract tumor, especially a prostate tumor; especially where such a tumor is refractory to treatment with another chemotherapeutic agent, especially one of the taxane class, more especially TAXOL® (preferably a hormone refractory prostate tumor); where epothilone B is administered weekly to a warm-blooded animal in a dose that is below 80%, more preferably below 50% of the maximum tolerated dose (MTD).

Preferably, in the case of treatment Weekly of a human with said epothilone (s), the dose is in the range of about 1 to about 60%, preferably about 10 to about 60%, for example about 5 to approximately 35% of BAT, for example in the range of about 30 to about 35% of BAT. Preferably for epothilone B the dose is in the range of about 5 to about 60%, preferably of about 10 to about 60%, especially in the range from about 10 to about 45%, more especially in the range of about 30 to about 45% of BAT every three weeks. In a special case, the dose may be between about 2 and about 18 mg / m2 for epothilone B.

More preferably, the treatment stops after the third to the eighth, especially after the third to the fifth weekly administration followed by a period of break two to five, for example two weeks before it Resume additional treatment. Preferably and if required, in the case of the weekly administration of epothilone B stops after the third to eighth administration followed by a rest period of two to four, for example two weeks before treatment is resumed by weekly administration

(6) The application describes the in vivo treatment of a tumor disease by combined administration (a) of epothilone B, in combination with (b) another chemotherapeutic agent selected from the group consisting of

(TO)

alkylating agents, preferably agents cross-linking chemotherapeutics, preferably bis-alkylating agents,

(B)

antitumor antibiotics, preferably the doxorubicin (ADRIAMYCIN®, RUBEX®);

(C)

antimetabolites;

(D)

plant alkaloids;

(AND)

hormonal agents and antagonists,

(F)

biological response modifiers, preferably lymphokines or interferons

(G)

protein tyrosine kinase inhibitors and / or serine / threonine kinases;

(H)

antisense oligonucleotides or derivatives of oligonucleotides; or

(I)

various agents or agents with another mechanism of action or unknown one;

quad

the combined treatment being planned so that component (a) and (b) are combined for simultaneous use or chronologically progressive within a period of time that is small enough for the active compounds both of the component (a) and component (b) mutually intensify the antiproliferative activity, for example in a patient.

(7) This application describes a product that it comprises a component (a) and a component (b) as defined with prior to (6), in the presence or absence of one or more pharmaceutically acceptable excipient materials, such as a combined preparation for simultaneous administration or chronologically progressive to a human within a period of time that is small enough for the compounds assets of both component (a) and component (b) intensify each other's activity against a tumor disease, especially (i) a tumor of the gastrointestinal tract, more especially a colon and / or rectum tumor (colorectal tumor); I (ii) a genitourinary tract tumor, especially a tumor of prostate; especially where such a tumor is refractory to treatment with another chemotherapeutic agent, especially one of the taxane class, more especially TAXOL®; for the treatment of said tumor disease.

From (1) to (7) or subsequent embodiments of the invention, the administration of epothilone B, is effected preferably by infusion, especially by infusion intravenous

The following are some realizations Especially preferred of the invention:

A1.

The use of epothilone B, for treatment of a proliferative disease that is refractory to treatment with another chemotherapeutic agent; or the use of epothilone B, for the preparation of a pharmaceutical preparation for the treatment of proliferative disease that is refractory to treatment with other agents Chemotherapeutic

A2.

The use according to A1 where the disease Proliferative is a tumor disease that is refractory to a microtubule stabilizing agent of the taxane class, especially TAXOL®.

A3.

The use according to any of A1 to A2 where the proliferative disease is a colorectal tumor, and / or a metastasis of this one.

A4.

The use according to any of A1 to A2 where the proliferative disease is a prostate tumor, and / or a metastasis thereof; especially a refractory prostate tumor to hormone.

B1.

The use of epothilone B for processing of a pharmaceutical preparation that is appropriate for the administration of said epothilone once in a dose that is between about 1 and about 100% of the maximum dose tolerated (BAT) in a warm-blooded animal, to said animal of warm blood for the treatment of a proliferative disease which is refractory to treatment with other agents Chemotherapeutic

B2.

The use according to B1 where epothilone it's epothilone B.

B3

The use according to any of B1 and B2 where the dose is between 25 and 100% of the maximum tolerated dose and the Warm-blooded animal is a human.

B4

The use according to any of B1 to B3 where the dose unit for an adult human is in the range between about 0.3 and about 18, preferably between about 0.3 and about 12, plus preferably between about 0.3 and about 7.5, more preferably between about 1.0 and about 3 mg / m2 of epothilone B with treatment every three weeks; or between about 0.1 and about 6, preferably about 0.1 to about 5, more preferably about 0.1 and about 3, more preferably about 0.3 and about 1 mg / m2 for the weekly treatment

B5

The use according to any of B1 to B4 where the dose is chosen such that after a period of time that allows sufficient recovery of the treated individual of each previous dose one dose can be administered additional.

B6

The use according to any of the B1 a B5 where proliferative disease is a tumor.

B7

The use according to any of B1 to B2 where the proliferative disease is a colorectal tumor, and / or a metastasis of this one.

B8

The use according to any of B1 to B6 where the proliferative disease is a prostate tumor, and / or a metastasis of this one.

B9

The use according to any of B1 to B8 where the tumor is one that is refractory to treatment with a microtubule stabilizing agent of the taxane class, especially TAXOL®.

C1.

The use of epothilone B, for preparation of a pharmaceutical preparation that is appropriate for the administration of said epothilone once a week, where the dose is 80% or less, preferably 50% or less, of the MTD

D1

The use of epothilone B, for preparation of a pharmaceutical preparation that is appropriate for the combined administration of an epothilone, preferably epothilone A and / or epothilone B, in combination with (b) another agent antitumor chemotherapeutic to a warm-blooded animal that suffer from a proliferative disease that is refractory to treatment with other chemotherapeutic agents, especially a colorectal or prostate tumor and / or a metastasis of East.

E1.

A pharmaceutical preparation for the treatment of a proliferative disease, especially a tumor disease, especially one of those characterized as preferred with before or after, in a human, said preparation that it comprises an epothilone, especially epothilone B, in a dose that It is in the range of 1 to 100%, preferably 25 to 100% of the maximum tolerated dose (MTD), and a pharmaceutically excipient  acceptable.

F1

A combined preparation comprising (a) epothilone A or preferably epothilone B and (b) one or more agents antitumor chemotherapeutics, and a pharmaceutically excipient acceptable.

G1.

A product that comprises as component (a) epothilone A and / or B, preferably epothilone B, and as a component (b) any other antitumor chemotherapeutic agent, in the presence or absence of one or more excipient materials pharmaceutically acceptable, as a combined preparation for administration simultaneously or chronologically progressive to a warm-blooded animal, especially a human, within a period of time that is small enough for the active compounds of both component (a) and component (b) mutually increase the antitumor activity in said animal of warm blood, for the treatment of a disease proliferative

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This application describes the treatment of following types of tumors / cancer with epothilone B:

(i)

a gastrointestinal tumor, especially a tumor colorectal that is refractory to a representative of the class of taxane of anticancer agents, in particular TAXOL®; or more especially to treatment with standard chemotherapy, especially with 5-fluorouracil, and / or TAXOL®.

(ii)

a genitourinary tract tumor, especially a prostate tumor, including primary tumors and especially metastatic; more especially if they are refractory to treatment hormonal;

(iii)

an epidermoid tumor, more especially a head and neck epidermoid, more especially an epidermoid of mouth, especially one of these that are refractory to treatment with another chemotherapeutic agent, especially due to the multi-drug resistance, especially treatment with a member of the taxane class of anticancer agents, especially TAXOL®;

(iv)

a lung tumor, especially a cancer of non-small cell lung, which is refractory to treatment with other chemotherapeutic agent, especially due to (mainly) resistance to multiple drugs, especially to treatment with a member of the agent taxane class anticancer, especially TAXOL®; I

(v)

a breast tumor, especially a breast tumor that it is resistant to multiple drugs, more especially one that is refractory to treatment with a member of the taxane class of anticancer agents, especially TAXOL®.

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Preferably, the invention refers to the treatment of any of the types (i) to (v) of tumors mentioned above, more preferably those of (I), (ii), (iv) and (v).

More preferably, the invention makes reference to the treatment of any of the types of tumors above mentioned under (i) to (v), especially to any of them, by treating with an intravenous infusion of epothilone B for 2 to 120 minutes, preferably during about 5 to about 30 minutes, more preferably for about 10 to about 30 minutes, more preferably for about 30 minutes; being told repeated administration every one to three weeks, preferably every week (weekly) or every three weeks; where the dose of epothilone B is preferably defined by formula I,

10

where N (an integer or fraction) is the number of weeks between treatments that is between about one to about three weeks, that is, N is from about 1 to about 3; e y is 6, preferably 5, more preferably Four.

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More preferably, the treatment dose is Calculates according to formula II

eleven

even more preferably of according to the formula III,

12

or more preferably agree with the formula IV

13

where, in each of the formulas from II to IV, N corresponds to between approximately 1 and approximately 3 (which preferably means from a weekly treatment until one treatment every three weeks)

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More preferably, for the weekly treatment the dose is between about 0.1 and about 6, preferably between about 0.1 and about 5 mg / m2, more preferably between about 0.1 and about 3 mg / m2, even more preferably between about 0.1 and about 1.7 mg / m2, plus preferably between about 0.3 and about 1 mg / m2; or for treatment every three weeks between about 0.3 and about 18 mg / m2, preferably between about 0.3 and about 15 mg / m2, plus preferably between about 0.3 and about 12 mg / m2, even more preferably between about 0.3 and approximately 7.5 mg / m2, even more preferably between about 0.3 and about 5 mg / m2, more preferably between about 1.0 and about 3.0 mg / m2.

More preferably, they may be necessary. rest periods of more than one week, more preferably of two to ten weeks, more preferably three to six weeks after the previous treatment (especially in the case of weekly treatment) after for example 3, 4, 6, 8, or more cycles, depending on the patient's condition, to allow sufficient recovery from previous treatment.

Also especially preferred are conditions and treatment formulations similar to those mentioned in the Examples.

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Pharmaceutical Formulations

This application describes the use of the epothilone B, for the preparation of a pharmaceutical formulation for use against a tumor disease as defined previously; or for a pharmaceutical formulation for the treatment of said tumor disease comprising epothilone B, and a pharmaceutically acceptable excipient.

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This application describes the compositions pharmaceuticals comprising epothilone B, for the treatment of a proliferative disease, especially a tumor disease defined as previously preferred, and to the preparation of Pharmaceutical preparations for said treatment.

Epothilone B can be used, for example, for the preparation of pharmaceutical compositions comprising an effective amount of active ingredients together with or in a mixture with a significant amount of inorganic excipients or organic, solid or liquid, pharmaceutically acceptable.

This application describes a composition pharmaceutical that is suitable for administration to an animal of warm blood, especially a human, for the treatment of a proliferative disease as defined earlier in this document, which comprises epothilone B, which is effective for the treatment of said proliferative disease, together with less a pharmaceutically acceptable excipient.

Pharmaceutical compositions according to the invention are those for enteral administration, such as nasal, rectal or oral, or preferably parenteral, such as intramuscular or intravenous to a warm-blooded animal (human or animal), which comprises an effective dose of the ingredient pharmacologically active, alone or together with an amount Significant pharmaceutically acceptable excipient. The dose of the active ingredient depends on the species of blood animal hot, body weight, age and individual condition, individual pharmacokinetic data, the disease that will be treated and the mode of administration; preferably, the dose is one of the preferred doses as defined above, which is appropriately modified if it is a treatment pediatric.

Pharmaceutical compositions comprise from about 0.00002 to about 95%, especially (for example in the case of infusion dilutions that are ready to be used) from 0.0001 to 0.02% or (for example in the case of infusion concentrates) from about 0.1% to about 95%, preferably from about 20% to approximately 90% active ingredient (weight / weight, in each case). The pharmaceutical compositions according to the invention they can be, for example, in unit dose form, such as in form of blisters, vials, suppositories, dragees, pills or capsules

Preferably, the dose is chosen so that allows the treatment regimen based on the BAT mentioned formerly for the sole or rare treatment of a disease tumor.

The pharmaceutical compositions of the present invention are prepared in a manner known per se , for example by means of conventional processes of dissolution, lyophilization, mixing, granulation or confection.

The solutions of the active ingredient, and also the suspensions, and especially the isotonic aqueous solutions or suspensions, are preferably used, this being possible, for example in the case of lyophilized compositions comprising the active ingredient alone or together with a pharmaceutically acceptable excipient, for example mannitol, for such solutions or suspensions to be produced before being used. The pharmaceutical compositions may be sterilized and / or may comprise excipients, for example preservatives, stabilizers, humectants and / or emulsifiers, solubilizers, salts for the regulation of osmotic pressure and / or buffers, and are prepared in a manner known per se , for example by means of conventional dissolution or lyophilization processes. Said solutions or suspensions may comprise substances that increase viscosity, such as sodium carboxymethyl cellulose, carboxymethyl cellulose, dextran, polyvinylpyrrolidone or gelatin.

Oil suspensions comprise as the oil component vegetable, synthetic or semi-synthetic habitual for the purposes of injection. It is possible to mention as such especially esters fatty acid liquids that contain as the acid component a long chain fatty acid that has 8 to 22, especially of 12 to 22, carbon atoms, for example lauric acid, acid tridecyl, myristic acid, pentadecyl acid, palmitic acid, margaric acid, stearic acid, arachidic acid, behenic acid or the corresponding unsaturated acids, for example oleic acid, elaidic acid, erucic acid, carbonic acid or linoleic acid, if desired with the addition of antioxidants, for example vitamin E, β-carotene or 3,5-di-tert-butyl-4-hydroxytoluene.  The alcohol compound of those fatty acid esters has a maximum of 6 carbon atoms and is a mono- or poly-hydroxy, for example a mono-, di- or tri-hydroxy, alcohol, for example methanol, ethanol, propanol, butanol or pentanol or its isomers, but especially glycol and glycerol. Therefore, the following should be mentioned Examples of fatty acid esters: ethyl oleate, isopropyl myristate, isopropyl palmitate, "Labrafil M 2375" (trioleate polyoxyethylene glycerol, Gattefossé, Paris), "Miglyol 812" (saturated fatty acid triglyceride with a chain length of C_ {8} to C_ {12}, Hüls AG, Germany), but especially oils Vegetables, such as cottonseed oil, oil almond, olive oil, castor oil, sesame oil, soybean oil and more especially peanut oil.

Injection or infusion compositions are prepare as usual under sterile conditions; the same It also applies to the introduction of compositions within blisters or ampoules and container sealing.

An infusion formulation is preferred which comprises epothilone B, and a pharmaceutically organic solvent acceptable.

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The formulation does not require the use of a surfactant Surfactants such as Cremophor can cause allergic reactions and can also filter plasticizers from Standard PVC containers, tubes and the like. In consecuense, when these are used, the use of devices of special infusion, for example tubes nitro-glycerin and non-plasticized containers, such as glass tubes and the like.

The pharmaceutically acceptable solvent used in a formulation according to the invention can be choose from any such organic solvent known in the art. Preferably the solvent is selected from alcohol, for example absolute ethanol or ethanol / water mixtures, more preferably 70% ethanol, polyethylene glycol 300, glycol polyethylene 400, polypropylene glycol or N-methylpyrrolidone, more preferably glycol of polypropylene or 70% ethanol or especially polyethylene glycol 300

Epothilone B may preferably be present in the formulation at a concentration of approximately 0.1 to about 100 mg / ml, more preferably of about 1 to about 100 mg / ml, even more preferably from about 1 to about 10 mg / ml (especially in infusion concentrates).

Epothilone B can be used as a substance pure, the use of an Epothilone A concentration of 5 is preferred at 100 mg / ml, preferably 10 a when Epothilone B is used in its pure form is preferably used in a concentration of 0.1 to 10, more preferably 1 to 10, even more preferably 1 to 2 mg / ml (this number refers especially to a concentrate of infusion that, before treatment, is diluted so corresponding, see below).

Such formulations are stored in a manner Convenient in ampoules or ampules. Usually the blisters or ampoules are made of glass, for example borosilicate glass or of soda and lime. Blisters or blisters can be of any conventional volume in the art, preferably they are of a size large enough to contain 0.5 to 5 ml of formulation. The formulation is stable for storage periods of up to 12 to 24 months at temperatures of at least 2 to 8ºC.

The formulations must be diluted in a aqueous medium suitable for intravenous administration before that epothilone can be administered to a patient.

The infusion solution should preferably have the same or essentially the same osmotic pressure as the body fluid. Therefore, the aqueous medium preferably contains an isotonic agent that has the effect of making the osmotic pressure of the infusion solution the same or essentially the same as that of the fluid.
bodily.

The isotonic agent can be selected from any of those known in the art, for example mannitol, dextrose, glucose and sodium chloride. Preferably the agent Isotonic is glucose or sodium chloride. Isotonic agents they can be used in amounts that grant the same or essentially the same osmotic pressure of the body fluid to the solution of infusion. The precise amounts needed can be determined through routine experimentation and will depend on the composition of the infusion solution and the nature of the agent isotonic The selection of a particular isotonic agent is performed based on the properties of the active agent.

The concentration of isotonic agent in the medium aqueous will depend on the nature of the specific isotonic agent used. When glucose is used, it is preferably used in a concentration of between 1 and 5% w / v, more particularly 5% w / v. When the isotonic agent is sodium chloride, it is used preferably in amounts of up to 1% w / v, in particular 0.9% p / v.

The infusion formulation can be diluted with the aqueous medium. The amount of aqueous medium used as a diluent is chosen according to the desired concentration of Epothilone in the infusion solution. Preferably the solution Infusion is done by mixing a blister or blister of the aforementioned infusion concentrate with an aqueous medium, increasing the volume to between 20 ml and 200 ml, preferably between about 50 and about 100 ml, with the medium aqueous.

Infusion solutions may contain other excipients commonly used in formulations that are administered intravenously. Excipients include antioxidants

Antioxidants can be used to protect Epothilone against oxidative degradation. Antioxidants can be chosen from any of those known antioxidants in the art and suitable for intravenous formulations. The amount of antioxidant can be determined by the routine experimentation As an alternative to adding a antioxidant, or in addition to that, the effect can be achieved antioxidant by displacing oxygen (air) from the contact with the infusion solution. This can be done by convenient way by purging the container it contains said solution with an inert gas, for example nitrogen.

Infusion solutions can be prepared by mixing a vial or vial of the formulation with an aqueous medium, for example a 5% w / v glucose solution in API or especially a 0.9% solution of sodium chloride in a suitable container, for example a bag or bottle of infusion.

The infusion solution, once formed, is preferably used immediately or shortly after being formed, for example within 6 hours.

The containers to house the solutions of infusion can be chosen from any conventional container that Do not be reactive with the infusion solution. The containers of glass made from those types of glass mentioned with previously they are suitable although the use may be preferred plastic containers, for example infusion bags plastic.

Plastic containers can be mainly those thermoplastic polymer compounds. The plastic materials may additionally comprise additives, by example plasticizers, fillers, antioxidants, antistatics and other conventional additives in the art.

Plastics suitable for this invention should be resistant to high temperatures required for thermal sterilization. Infusion bags Preferred plastics are those made from materials PVC plastics known in the art.

A wide range of sizes of containers When container size is selected, it must be consider the solubility of epothilone in the aqueous medium and the ease of handling and, if appropriate, storage of container.

It is preferred to use containers that they can hold between approximately 250 and 1000 ml of solution of infusion, but preferably between about 50 and approximately 120 ml

Infusion solutions act in a way similar to the infusion solutions of paclitaxel agents that interact with microtubules, and are beneficial in treatment of diseases for which paclitaxel could be used. For certain tumors, epothilones offer beneficial effects. boosted compared to paclitaxel.

The dosage forms can be conveniently administered intravenously at a dose of up to about 18 mg / m2 of Epothilone B. The exact dose required and the duration of administration will depend on the severity of the disease and the rate. of administration, and is preferably as defined above. Since the dose can be delivered intravenously, the dose received and blood concentration can be determined accurately based on known in vivo and in vitro techniques.

Pharmaceutical compositions for oral administration can be obtained by combining active ingredients with solid excipients, if desired by the granulation of a resulting mixture, and the processing of the mixing, if desired or necessary, after the addition of appropriate excipients, in the form of pills, dragee cores or capsules It is also possible that they are incorporated into plastic excipients that allow active ingredients to disseminate or be released in measured quantities.

The pharmaceutically acceptable excipients Suitable fillers are especially suitable, such as sugars, for example lactose, sucrose, mannitol or sorbitol, cellulose and / or calcium phosphate preparations, for example tricalcium phosphate or calcium hydrogen phosphate, and binders, such as the starch pastes they use for example corn, wheat, rice or potato starch, jelly, gum tragacanth, methylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose sodium and / or polyvinylpyrrolidone, and / or, if desired, disintegrators, such as the aforementioned starches, also carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate. The excipients are especially flow conditioners and lubricants, for example silicic acid, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and / or polyethylene glycol. The Dragee cores are provided with suitable coatings, optionally enteric, which are in use, among others, solutions of concentrated sugars that may comprise gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, or coating solutions in suitable organic solvents, or, for the preparation of enteric coatings, solutions of suitable cellulose preparations, such as phthalate ethyl cellulose or hydroxypropylmethyl cellulose phthalate. Capsules they are hard capsules made of gelatin and soft sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Hard capsules may comprise the active ingredient. in the form of granules, for example with fillers, such as lactose, binders, such as starch, and / or glidant, such as talc or magnesium stearate, and if desired with stabilizers. In soft capsules the solution is preferably dissolved or suspended. active ingredient in suitable oily excipients, such as fatty oils, paraffin oil or polyethylene glycols liquid, and stabilizers and / or agents can also be added antibacterials Dyes or pigments can be added to coatings of the tablets or dragees or the wrapping of the capsules, for example for identification purposes or to indicate Different doses of active ingredient.

In the case of combinations with others chemotherapeutic agents, a fixed combination of two or more components (a) and (b) as defined above or two or more independent formulations (for example in a set of parts) are prepared as described above, or the other (s) chemotherapeutic agent (s) is used in standard formulations that are marketed and are known by the art experts.

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Examples

The following examples are intended for illustrate the present invention but are not intended to limit its scope. Especially the cell lines mentioned there are not intended limit the scope of the invention since they are merely representative and can be replaced with different lines cell and tumor cells of which they are representative.

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Preparation of compound solutions

A stock solution of epothilone B a is prepared 10 mg / ml in DMSO and stored at -20 ° C. The aliquots are diluted in aqueous solutions in a final concentration of 5% v / v DMSO, of Tween 80 (polyoxyethylene sorbitan monooleate; ICI Americas, Inc.) 0.05% v / v, and 95% v / v physiological serum (0.9% w / v NaCl).

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Conditions of cells and cell culture

The HCT-15 (ATCC CCL 225) cell line of human colorectal adenocarcinoma belongs to the American Type Culture Collection (Rockville, MD, USA), and the cells are cultured in vitro as recommended by the provider. HCT-12 is an epithelial cell line (Cancer Res. 39 1020-25 [1979]) that is resistant to multiple drugs due to the overexpression of P-glycoprotein (P-gp, gp170, MDR-1; Anticancer Res. 11 1309-12; J. Biol. Chem. 264: 18031-40 [1989]; Int. J. Cancer 1991; 49: 696-703 [1991]) and glutathione-dependent resistance mechanisms (Int. J. Cancer 1991; 49: 688-95. [1991]).

The Colo 205 cell line is also a line cell carcinoma of the human colon (ATCC No. CCL 222; see also Cancer Res. 38, 1345-55 [1978] which was isolated from a patient's ascites fluid, sample epithelial type morphology and is considered generally sensitive to the drugs.

A human cancer cell line of androgen independent prostate to establish models subcutaneous and orthotopic in mice. Prostate carcinoma PC-3M human metastatic is obtained from Dr. I.J. Fidler (MD Anderson Cancer Center, Houston, TX, USA) and se cultivate in Ham F12K media supplemented with 7% v / v FBS. The PC-3M cell line is the result of isolation from liver metastases in nude mice after intrasplenic injection of PC-3 cells [ATCC CRL 1435; American Type Culture Collection (Rockville, MD, USA) UU.)], And can grow in Eagle MEM supplemented with 10% serum  fetal bovine, sodium pyruvate, non-essential amino acids, L-Glutamine, a double vitamin solution (Gibco Laboratories, Long Island, NY.) And penicillin-streptomycin (Flow Laboratories, Rockville, Md.). The PC-3M cell line is hormone insensitive (i.e. it grows in the absence of androgens). The PC-3 cell line is a receptor for negative androgens, just as the line is presumably PC-3M derived cell. PC-3 is a cell line available in ATCC (ATCC CRL 1435) and corresponds to a grade IV prostatic adenocarcinoma isolated from a 62-year-old Caucasian male; cells exhibit low acid phosphatase activity and testosterone-5-α-reductase. The cells are almost triploid with a modal number of 62 chromosomes Abnormal Y chromosomes can be detected by Q band analysis.

A549 human lung adenocarcinoma (ATCC CCL 185; isolated as explant culture from a tissue of lung carcinoma of a 58-year-old Caucasian male); shows epithelial morphology and can synthesize lecithin with a high percentage of unsaturated fatty acids that use the route citidine diphosphocholine; is in all metaphases a subtelocentric marker chromosome involving a chromosome 6 and the long arm of a chromosome 1. Human breast carcinoma ZR-75-1 (ATCC CRL 1500; isolated a from a malignant ascitic effusion of a caucasian woman of 63 years old with infiltrating ductal carcinoma); Is from origin mammary epithelial; the cells have estrogen receptors and other steroid hormones and have a number of chromosomes hypertriploids

The KB-8511 human epidermal (mouth) carcinoma cell line (an overexpression cell line derived from a KB-31 epidermoid (mouth) carcinoma cell line) is obtained from Dr. RM Baker, Roswell Park Memorial Institute (Buffalo , NY., USA) (for a description see Akiyama et al ., Somat. Cell. Mol. Genetics 11, 117-126 (1985) and Fojo A., et al ., Cancer Res. 45, 3002- 3007 (1985)) and grown as described above (Meyer, T., et al ., Int. J. Cancer 43, 851-856 (1989)). KB-8511 cells, such as KB-31, are derived from the KB cell line (ATCC) and are human epidermal carcinoma cells; KB-31 cells can be cultured in monolayer using a modified Dulbecco Eagle (D-MEM) medium with 10% fetal bovine serum (MA Bioproducts), L-glutamine (Flow), penicillin (50 units / ml) and streptomycin (50 µg / ml) (Flow); then they grow with a doubling time of 22 hours, and their relative efficiency of plate culture is approximately 60%. KB-8511 is a cell line derived from the KB-31 cell line through the use of colchicine treatment cycles; shows a relative resistance 40 times higher against colchicine when compared to KB-31 cells; It can be grown under the same conditions as KB-31.

For more details about the features of cell lines, see the ATCC catalog and references cited there, or the other references cited above.

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The following cell lines mentioned with previously they have been deposited under the Budapest Treaty on 20 February 1998, at the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Mascheroder Weg 1b, D-38124 Braunschweig, Germany) under the following accession numbers, respectively: PC-3M: DSM ACC2338; A-549: DSM ACC2337; KB-8511: DSM ACC2342.

In addition, the following cell lines mentioned above have been deposited under the Treaty of Budapest on December 1, 1998 at Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Mascheroder Weg 1b, D-38124 Braunschweig, Germany) under the following accession numbers, respectively: ZR-75-1: DSM ACC2376; HCT-15: ACC2377.

The methods are described below. General for the tests performed. Where they are mentioned special conditions, these prevail over the descriptions generals presented in the following paragraphs:

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Antiproliferative assays

Antiproliferative tests are performed as was described previously (Int. J. Cancer 43, 851-6 (1989)). Briefly, cells are seeded 1.5 x 10 3 cells / well in 96-well microtiter plates and incubate overnight. Compounds are added in serial dilutions on day 1. The plates are then incubated for 2 to 5 additional days, allowing at least one duplication cell (depending on the cell line), after which the cells are fixed with 3.3% v / v glutaraldehyde, washed with water and stained with 0.05% w / v methylene blue. After washing, it dilute the dye with 3% v / v HCl and the optical density is measured at 665nm with a SpectraMax 340 (Bucherer, Basel, Switzerland). The values IC50 are determined by a computerized system (SoftPro, Bucherer, Basel, Switzerland) using the formula (OD test - OD start) / (OD control - OD start) x 100. The IC50 value is defined as the concentration of drug that leads to 50% of cells per well compared to control crops (100%) at the end of the period of incubation.

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In vivo antitumor activity against sc transplanted tumors

BALB / c nu / nu (nude) female mice or males are kept under sterile conditions (from 10 to 12 Type III cage mice) with free access to food and water. The mice weigh between 20 and 25 grams at the time of implantation of the tumor. Tumors are established by subcutaneous injection of cells (minimum 2 x 10 6 cells in 100 µl PBS or medium) in carrier mice (4-8 mice per cell line). The resulting tumors are subjected to serial passage for a minimum of three consecutive transplants before starting treatment. Tumor fragments (approximately 25 mg) are implant s.c. within the left flank of the animals with a 13 gauge trocar needle while mice are exposed to Forene anesthesia (Abbott, Switzerland).

The growth of the tumor and the body weights once or twice a week. All treatments administered intravenously (i.v.) and started when reaches an average tumor volume of approximately 100 to 250 mm3, depending on the type of tumor. Tumor volumes are determined using the formula (L x D x \ pi / 6 (see Cancer Chemother Pharmacol 24: 148-154, [1989]). The Epothilone B treatments vary the dose and frequency of administration. The comparison agents are administered according with optional treatment regimens previously determined. In addition to presenting changes in tumor volumes throughout the treatment, the antitumor activity is expressed as T / C% (average increase in tumor volumes of animals treated divided by the average increase in volumes tumor of control animals multiplied by 100). The tumor regression (%) represents the average tumor volume plus small compared to the average tumor volume at the beginning of treatment, according to the Regression formula (%) = (1 - V_ {final} / V_ {beginning}) x 100 (V_ {end} = tumor volume final average, V_ {beginning} = average tumor volume at beginning of treatment Every animal is sacrificed in which the tumor has reached a size that exceeds approximately 1.5 to 2.5 cm3. Details can be found below.

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In vivo antitumor activity against orthotopically injected cells

PC-3M cells of human prostate carcinoma (1 x 10 6 cells per 20 µl phosphate buffered saline) are injected into the left ventricle of each animal's prostate (n = 6-9 / group ) according to the methods described above (see Stephenson et al ., J. Natl. Cancer Inst. 84, 951-7 (1992)). Treatment begins on day 14 after cell injection. At this time an average tumor weight of -20 mg is established. Epothilone B is given once or once a week. Mice are sacrificed 42 days after tumor inoculation, and prostates are carefully removed, adherent tissues are dissected and weighed. Mesangial lymph nodes are examined for the presence of metastases, adherent tissues are dissected and weighed. The mesenteric lymph nodes are examined for the presence of metastases, the adherent tissues are dissected and weighed.

Tumor weight and weights are monitored body once or twice a week. All treatments are administered intravenously (i.v.) and started 14 days later of cell injection. Treatment with epothilone B varies the dose and frequency of administration. The antitumor activity is expressed as T / V% (the average tumor weight of the animals treated divided by the average tumor weight of the animals of control multiplied by 100).

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Statistical analysis

The basic strategy for analysis statistical is to use tests for multiple comparisons to judge the statistical relevance of the differences between treatment groups, and differences within a group (it is say, compared to the beginning of treatment) to determine if the treatment induces stable regressions of the disease or tumor Since subcutaneous tumor volumes the differences in volumes are not normally distributed Subcutaneous tumors between treatment groups are determined using the ANOVA test of a non-parametric address of Kruskal-Wallis on distributed data (Proof of Sum of Ranks), and the statistical relevance of the differences between the treatment groups compared with the groups of control determined using the Dunnett test. The comparisons in pairs between all groups is done using the method Student-Newman-Keuls (SNK). The organ weights are not always distributed normally and are analyzed using non-parametric tests (as shown above) or they are transformed to normal (Log [weight of organ]) and analyzed by unidirectional test ANOVA Dunnett (comparison to controls) or using the test Tukey (comparisons between groups). The statistical analysis on tumor volumes \ Delta uses the ANOVA test unidirectional Kruskal-Wallis on data distributed, comparing treatment groups with controls vehicles using the Dunnett test. The differences in body weights between treatment groups with controls vehicles are analyzed by paired t tests. The proof Fisher's exact is used to determine the relevance of differences in mortality between groups. For all tests the level of relevance is set at p <0.05, but note that For these small sample sizes, you never get the level of desired power of 0.8. For the orthotopic model, the Fisher's exact test to determine if the proportions of Mice that undergo metastases are different between groups of Treatment and control. All statistical calculation is performed using SigmaStat 2.0 (Jandel Scientific).

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Material

Epothilone B is purified from cultures of the Sorangium cellulosum myxobacterium by the Department of Biopharmaceutical Development and Productions, Novartis Pharma, Basel, Switzerland. TAXOL® (paclitaxel formulated for clinical use) purchased from Bristol Myers Squibb (United States), paclitaxel a Calbiochem (United States), and 5-fluorouracil (Fluorouracil®) is from Roche (Switzerland). Crop materials Cell phones are from Integra BioSciences (Wallisellen, Switzerland). The media liquids, fetal bovine serum (FBS) and media additives are from Gibco / BRL (Basel, Switzerland). BALB / c nu / nu (nude) mice are from Bomholtgaard (Copenhagen, Denmark) or are obtained from Novartis Animal Farm (Sisseln, Switzerland). Normal BALB / c mice are from Iffa Credo (France) or are obtained from Novartis Animal Farm (Sisseln, Switzerland).

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Determination of the maximum tolerated dose (BAT)

For the determination of BAT, the female nude BALB / c mice or BALB / c mice (obtained from Novartis Animal Farm, Sisseln, Switzerland) once intravenously with epothilone B (n = 3 per dose group). The dose is increased (2, 4, 6, 8, 10 and 12 mg / kg) and the mice are observed in search of the presentation of obvious toxic effects for 10 days after of drug treatment.

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Example 1 Determination of the maximum tolerated dose (BAT)

The results from the dose study Maximum tolerated (BAT) are presented in Table 1 and Table 2.

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TABLE 1 BAT determination of the single intravenous dose for epothilone B in female BALB / c mice normal

Body weight (average g \ pm SD)

14

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Epothilone B is given as a bolus dose single intravenous 2, 4, 6, 8, 10 or 12 mg / kg. The Survival and body weight of mice daily. The Changes (Δ) in body weights are determined by comparing the last body weight measured with body weight before treatment.

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TABLE 2 BAT determination of the single intravenous dose for epothilone B in female BALB / c mice nudes

Body weight (average g \ pm SD)

fifteen

Epothilone B is given as a bolus dose single intravenous 2, 4, 6, 8, 10 or 12 mg / kg. The Survival and body weights of mice daily. The Changes (Δ) in body weights are determined by comparing the last body weight measured with body weight before treatment.

From these experiments it follows that in normal mice the MTD is approximately 4 mg / kg, while in The naked mice the MTD is approximately 6 mg / kg.

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Example 2 Toxicity of Epothilone B (Two week comparative study of intravenous toxicity in mice)

In order to assess intravenous toxicity subchronic epothilone B, a study of Two-week intravenous toxicity that does not comply with GLP in normal female BALB / c mice that have no tumor, which implies mortality analysis, clinical signs, body weight, consumption of food, hematology, clinical biochemistry, urinalysis, and weight of the organs as well as macro and microscopic exams. The study It is based on two different dose regimens of epothilone B of 3 mg / kg and 10 mg / kg, respectively, administered on days 1 and 8 (8 animals per group). Half of the animals are killed on day 15 (main group) and a necropsy is performed. For the other half (recovery group) a recovery period of 5 is allowed weeks after administration of the second dose before sacrifice and subsequent necropsy on day 43. However, for the 10 mg / kg dose had to be sacrificed to all animals in the group of recovery prematurely on day 19, due to commitment of the general state.

There was no mortality throughout the period of treatment at either dose level (days 1-14) and for the 3 mg / kg dose group all recovery group animals survived until the end of study. Body weight loss is observed for all animals in the dose group of 10 m / kg during the first and second weeks, while the loss of body weight in the dose of 3 mg / kg It is only visible in the second week. Weight development body during the recovery period is similar for treated and control animals.

Both dose levels of epothilone B induce a reduction in the number of leukocytes, especially of neutrophils and lymphocytes, in all treated animals (day 15), but the effect is more pronounced in the dose of 10 mg / kg. Further, slight increases in basophil count are observed as well also decreased levels of monocytes in some animals in particular at both dose levels. Values were recorded slightly lower in red blood cell parameters with increases in reticulocytes and platelets only for the dose of 10 mg / kg At the end of the recovery period (day 43) the hematological parameters are normal for two of the four animals of the dose of 3 mg / kg of the recovery group, while that the other two still suffer from reduced blood cell counts white.

Only minor changes in the profile were observed clinical chemist of treated animals, which cannot be associated clearly with epothilone B treatment.

Treatment with epothilone B at both levels of dose leads to pronounced changes in the weight of the thymus, spleen, and uterus (day 15). In addition, slight reductions in weight are observed of the liver The organ weight for the glands is determined adrenal, liver, thymus, spleen, brain, ovaries, the kidneys, the uterus, and the heart). For the dose of 3 mg / kg, the organ weights at the end of the recovery period (day 43) they are comparable with those of control animals, which indicates A total recovery. (Organ weights are not taken to animals slaughtered at the dose of 10 mg / kg of the group of Recovery).

The microscopic investigation of tissues Histologically processed selected from animals sacrificed on day 15 reveals moderate to marked thymus atrophy for the dose of 3 mg / kg and that of 10 mg / kg, respectively. Further, minimal lymphoid atrophy is observed in the spleen, myeloid hypoplasia minimum to light in the bone marrow of the sternum, and minimal increase in hemopoiesis in the spleen at the dose of 10 mg / kg. In the dose of 3 mg / kg the bone marrow of the sternum shows minimal to slight atrophy Erythroid and myeloid. Minimum single cell necrosis is detected in the intestinal mucosa (small and large intestine) in both levels of dose, but with a higher incidence in the dose of 10 mg / kg.

The animals of the recovery groups of both dose levels show mild myeloid hyperplasia and / or bone marrow atrophy. In the 10 mg / kg atrophy is also seen light lymphoid in the spleen and there is also presence of hemosiderosis Light to moderate There is no thymic tissue available for the test 10 mg / kg microscopic, indicating that thymus atrophy could be also present in these mice. They can not be found histological abnormalities in the thymus on day 43 for animals of the 3 mg / kg dose of the recovery group.

In conclusion, at the dose level of 3 mg / kg (dose on days 1 to 8) epothilone B is tolerated without mortality for the total observation period of 43 days, while the animals with doses of 10 mg / kg have to be slaughtered in the day 19, due to the commitment of the state of health. Weight loss Body occurs at both dose levels during treatment. The hematology reveals lower values for leukocytes, neutrophils and lymphocytes with higher basophil counts and lower monocytes for some mice in particular in both dose groups. In addition, evidence of anemia is seen at both levels of dose. No effects on the clinical chemical profile are observed. Be observe moderate to marked atrophy of the thymus at 3 and 10 mg / kg after of the treatment period only (day 15), along with atrophy minimum lymphoid in the spleen at the dose of 10 mg / kg. Also I know detects myeloid hypoplasia in the bone marrow and hemopoiesis increased in the spleen by 10 mg / kg. Erythroid and myeloid atrophy is seen Light bone marrow at 3 mg / kg. Cellular necrosis is detected unique in the intestinal mucosa at 3 and 10 mg / kg at the end of treatment only.

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Conclusions

The most important conclusions that emerge to from the data summarized in Examples 1 and 2 for Toxicological findings with epothilone B can be summarized as as follows:

BAT for the administration of a single dose intravenous epothilone B to normal and nude mice BALB / c corresponds to 4 mg / kg and 6 mg / kg, respectively. Therefore, the nude mice are less sensitive than normal mice to toxic effects of the compound.

In normal mice, they are tolerated reasonably well two doses of 3 mg / kg given with a range of one week and do not cause mortality until day 43 after initial dose The same dose regimen at the dose level of 10 mg / kg results in the death (or sacrifice) of all treated animals

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Example 3 In vitro activity of epothilones against cell lines

The powerful activity is confirmed antiproliferative epothilone B for some cell lines of human cancer; The results of these experiments are summarized in the Table 3. Epothilone B generally exhibits greater potency than the paclitaxel, particularly against cancer cells with a multi-drug resistance (MDR) phenotype (for example KB-8511, HCT-15).

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TABLE 3 In vitro activity of epothilones against human carcinoma cell lines

16

Example 4 Antitumor activity of epothilones against tumors of human colorectal HCT-15 adenocarcinoma

Tumor volumes are used as the primary indicators of the activity of antitumor agents used alone or in combination, and changes in the body weights as an indicator of tolerability at treatment.

As can be deduced from Table 4, a single dose of 4 mg / kg of epothilone B is capable of producing tumor regressions (p <0.05 vs. control vehicles; test of Dunnett) in drug-resistant HCT-15 tumors with overexpression of P-gp (Figure 1 and Table 1). This activity is clearly superior to five administrations of 20 mg / kg TAXOL® or two administrations of 75 mg / kg of 5-fluorouracil (p <0.05 vs, epothilone B; test SNK). HCT-15 tumors are resistant to both TAXOL® as with 5-fluorouracil, in which they are obtained the final T / C values of 50% and 88%, respectively (both p> 0.05 vs. controls; Dunnett's test). Epothilone treatment B is well tolerated in that the body weight is stable under treatment; treated vehicle mice gain weight. I dont know observes mortality due to treatment with epothilone B. Instead, some mortality is observed with TAXOL® treatment (1/8 [12.5%] deaths) and a higher degree of lethality occurs with the 5-fluorouracil (4/8 [50%] deaths); but nevertheless, Due to the small size of the treatment groups, this does not reaches statistical relevance (p> 0.05 vs. controls; Test Fisher's exact). Mice that survive any of the Two treatments demonstrate stable body weights.

This result indicates that epothilone B produces a pronounced antitumor effect against tumors HCT-15 resistant to TAXOL® and 5-fluorouracil and is well tolerated in this dose of 4 mg / kg

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TABLE 4 Antitumor effect of epothilone B in comparison with TAXOL® or 5-fluorouracil against carcinoma of human HCT-15 colon transplanted so subcutaneous in nude mice BALB / c female

18

Tumor fragments of approximately 25 mg are implanted inside the left flank of each naked mouse female (n = 8 per group). The treatments begin on day 14 after tumor transplantation. Epothilone B is administered once in a dose of 4 mg / kg, intravenously, on day 14. The 5-Fluorouracil is administered in doses of 75 mg / kg, intravenously on days 14 and 21. TAXOL® is administered in dose of 20 mg / kg / day, intravenously, every second day for 5 treatments (days 14, 16, 18, 20 and 22). The antitumor activity is expressed as T / C% (average increase in tumor volumes of treated animals divided by the average increase in tumor volumes of control animals multiplied by 100). Tumor regression (%) represents tumor volume final average compared to the average tumor volume at the beginning of treatment Δ Tumor volumes represent the tumor volume on the last day of treatment minus volume tumor on the first day of treatment. For the analyzes statistics of the Δ tumor volumes the Dunnett's test to compare treatment groups with those of control. For the statistical analysis of weight changes body paired t tests that compare weights are used body before treatment with body weights at the end of the treatment; mice weight -20-25 g per start of treatment Lack of relevance is indicated by the abbreviation "ns". The data presented are averages ± SEM from animals that survive at the end of the experiment.

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Example 5 Antitumor effect of epothilone B compared to the TAXOL® against squamous cell carcinoma KB-8511 human transplanted subcutaneously in nude mice BALB / c female

As can be deduced from Table 5, different epothilone B regimens are able to inhibit the KB-8511 tumor growth resistant to TAXOL® in nude mice. A single administration of 4 mg / kg of epothilone B produces a transient regression (- 51% on day 25 post transplantation; p <0.05 vs. control vehicles, Dunnett), but tumors grow back to the 40th day after to the treatment that results in a final T / C of 24% (p <0.05 vs. vehicle controls, Dunnett). This unique administration of epothilone B is well tolerated producing stable body weights, and without mortality.

Intravenous administration once per epothilone B week results in inhibition dependent on Tumor growth dose: 4 mg / kg produces regressions of 98% (p <0.05 vs. control vehicles; Dunnett); 2 mg / kg, produces Transient regressions of 44% and a final T / C of 14% (both p <0.05 vs. control vehicles; Dunnett); 1 mg / kg produces a T / C 81% final (p> 0.05 vs. control vehicles; Dunnett). He TAXOL® is inactive against KB-851 tumors (T / C 132%, p> 0.05 vs. control vehicles; Dunnett). In the end of the experiment, 5/8 mice treated with 4 mg / kg / week, and 1/8 Mice treated with 2 mg / kg / week of epothilone B have tumors undetectable. Although 4 mg / kg once a week tend to reduce body weights (-5 ± 7%), this is not relevant statistics. Control vehicles, 2 and 1 mg / kg / week of epothilone B, and TAXOL® groups all show body weights increasing, and there is no mortality, which suggests that Treatments are well tolerated.

These results indicate that epothilone B is effective against experimental epidermoid tumors that are TAXOL® resistant.

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TABLE 5 Antitumor effect of epothilone B in comparison with TAXOL® against squamous cell carcinoma KB-8511 human transplanted subcutaneously in nude mice BALB / c female

19

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Tumor fragments of approximately 25 mg are implanted inside the left flank of each naked mouse female (n = 8 per group). The treatments begin on day 13 after tumor transplantation. Epothilone B is given once in doses of 4 mg / kg, intravenously, on day 13, or a once a week in doses of 4, 2 or 1 mg / kg, intravenously, ( days 13, 21 and 27). TAXOL® is administered intravenously in dose of 20 mg / kg / day every second day for 5 treatments (days 13, 15, 17, 19 and 21). The antitumor activity is expressed as T / C% (average increase in animal tumor volumes treated divided by the average increase in tumor volumes of control animals multiplied by 100). Regression Tumor (%) represents the final average tumor volume compared with the average tumor volume at the beginning of treatment. The changes (Δ in tumor volumes represent volume tumor on the last day of treatment minus tumor volume in The first day of treatment. For statistical analyzes of Δ tumor volumes Dunnett's test is used to compare the treatment groups with the control groups. For the Statistical analyzes of body weight changes are used paired t tests comparing body weights before treatment with body weights at the end of treatment; weight of mice -20-25 g at the start of treatment. Lack of relevance is indicated by the abbreviation "ns". The Data presented are averages ± SEM of animals that survive at the end of the treatment

From this experiment it follows that, while the TAXOL® is not effective, treatment with epothilone B shows effective antitumor activity; you can find even regression with the dose of 4 mg / kg.

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Example 6 Antitumor activity of epothilones against the cells of PC-3M prostate injected orthotically

The results that determine the activity of the epothilone B against growing PC-3M tumors initially in the prostate and then form metastases in the mesangial lymph nodes are presented in Table 6.

In this experimental cancer model of prostate, PC-3M cells initially grow in the prostate and then form lymph node metastases mesenteric The weights of the organs are used to assess the antitumor activity of treatments.

Table 6 represents the progress of cancer experimental after 14 days after the injection of cells (start of treatment), where no involvement of lymph nodes, 42 days after the injection of cells, where the prostate and mesenteric lymph nodes They gain weight dramatically. All regimes of administration of epothilone B are highly effective (all p <0.05 versus controls; Dunnett's test on data logarithmically transformed) in reducing the weights of the prostate and prevent tumor metastasis to lymph nodes mesenteric All active treatments are equivalent in antitumor activity (p> 0.05; from Dunn). In each of the Epothilone B treatment groups, only one animal has detectable metastasis, compared to all animals in the treated vehicle controls (p <0.05; Fisher's Exact test), indicating that treatment with epothilone B reduces significantly the formation of detectable metastasis.

Epothilone B treatments are not good tolerated in higher doses. While an administration single 6 mg / kg, or two administrations of 4 mg / kg, only tend to reduce body weights, administration of 8 mg / kg once, or 5 mg / kg once a week, produce significant losses in body weight (Table 6). The treatment seems to promote survival of mice that have the tumor; but nevertheless, probably due to small numbers per group of treatment, this only reaches statistical relevance with the regimen 6 mg / kg (once) (p = 0.029, Fisher's Exact test on final survival numbers).

Epothilone B demonstrates excellent activity antitumor in this model, both in terms of reducing Primary tumors as prevention of metastasis. However, the Epothilone B, in some regimens, is not well tolerated.

The results of this study indicate that epothilone B is active against human prostate carcinomas both in vitro and in vivo (see Example 3). Epothilone B is able to reduce the growth of the primary tumor and potently inhibit the formation of detectable metastases in an orthotopic model of prostate cancer. In addition, it can also promote the survival of these mice that have the tumor, although this needs to be examined in additional experiments. Parallel to its potent antitumor activity, treatment with epothilone B causes significant body weight losses with the proven dose regimens. However, the reasons for this poor tolerability are unknown.

Epothilone B activity in the model Orthotopic PC-3M is especially notable. The Orthotopic models are designed to implant the tumor within the tissues where the primary tumor is located in humans, and to difference from most subcutaneous implantation models of tumors, metastases appear frequently. Therefore, the repression of the growth of the primary tumor in the prostate by epothilone B, and the inhibition of the formation and / or growth of Mesenteric lymph node metastasis represents a significant activity of epothilone B.

In short, due to the powerful activity antitumor in experimental prostate cancer models, which they are considered relatively resistant to chemotherapy (Br. J. Cancer 75, 1593-600 (1997)), epothilone B seems be a promising agent for the treatment of cancer prostate.

TABLE 6 Antitumor effect of epothilone B against cells of human PC-3M prostate carcinoma injected from orthotopic form in male nude BALB / c mice

twenty

1 x 10 6 PC-3M is injected cells in 20 µL of PBS inside the left ventricle of the Prostate of each male nude mouse (n = 9 per group). The treatments begin on day 14 after cell injection Tumor Epothilone B is given intravenously once in doses of 6 or 8 mg / kg., or once a week in doses of 4 or 5 mg / kg Tumor activity is expressed as T / C% (tumor weight average of treated animals divided by average weight tumor of control animals multiplied by 100). The differences in body weights consider only animals that survive until the end of the experiment (day 42). For the Statistical analyzes of Δ body weight tests are used t matched comparing body weights before treatment with body weights at the end of treatment; weight of mice? 20-25 g at the beginning of treatment. Lack of relevance is indicated by the abbreviation "ns."

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Example 7 Effect of epothilone B against lung carcinoma of non-small human cells A549

3-10 million cells subcutaneously. within the axillary (lateral) region right of atomic exogamic (nu / nu) mice, and allowed grow until a tumor volume of approximately 100 mm3. Epothilone B is formulated in 1% DMSO in 5% glucose in distilled water (D5W), and administered intravenously once only, once a week, 3 times a week, or 5 times per week. Positive controls are performed with the formulations TAXOL® clinics diluted 6 times more with D5W and administered by intravenous route 3x / week.

The antitumor activity is expressed as% T / C (comparing Δ tumor volumes for the group of treatment with the vehicle control group) at the end of experiment. Regressions are calculated using the formula: - (T / T_ {0} -1) x 100%, where T is the tumor volume for the group of treatment at the end of the experiment, and T 0 is the volume tumor at the beginning of the experiment. The relevance is evaluated statistics using a Student's t-test of a tail.

Results : Table 7 summarizes the results for A549 tumors. Epothilone B induces significant tumor inhibition (T / C = 41%), with undetectable toxicity, when administered once in doses of 6 mg / kg. A dose of 4 mg / kg administered 1x / week (4 mg / week) induces tumor stasis (T / C = 7%), but also produces a 13% body weight loss. In comparison, at a dose of 1.5 mg / kg administered 3x / week (4.5 mg / week) all animals have to be slaughtered in the first week of the experiment due to toxicity. A dose of 0.5 mg / kg, 5x / week (2.5 mg / week) induces a tumor inhibition identical to that of the once-only regimen (T / C = 41%), but the obvious cumulative toxicity gives as resulted in a weight loss of 23%. TAXOL®, administered 3x / week with a dose of 20 mg / kg does not inhibit tumor growth and induces a 16% body weight loss, with lethality in 1 of 8 mice.

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TABLE 7 Antitumor activity of Epothilone B, compared with TAXOL®, in non-small cell lung tumors A549 in nude mice

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The treatments begin on the 16th after implantation (10 million cells / animal). Epothilone B is given intravenously once at a dose of 6 mg / kg (day 16), once a week at a dose of 4 mg / kg (days 16, 23, and 30), three times a week at doses of 1.5 mg / kg (days 16, 18, 21, 23, 25, 28, 30, 32, and 35), or five times a week at 0.5 mg / kg (days 16-18, 21-25, 28-32, and 35-36). TAXOL® is administered intravenously three times per week in doses of 20 mg / kg (days 16, 18, 21, 23, 25, 28, 30, 32, and 35) in separate doses of 10 mg / kg given with An interval of one hour. The control vehicle (1% DMSO / D5W) is administered intravenously three times per week (days 16, 18, 21, 23, 25, 28, 30, 32, and 35). All final data is recorded on day 37. An asterisk only (*) indicates p <0.05, and a double asterisk (**) indicates p <0.01, using a single-tailed Student's t-test. "NE": not evaluable - animals slaughtered due to the toxicity of the compound.

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Example 8 Antitumor activity of epothilone B compared to TAXOL® in non-small cell lung tumors A549 in mice nudes

The materials and methods concerning the model Xenographic of the human tumor are as described previously. 10 or 1 million cells (A549) are implanted subcutaneously within the right axillary (lateral) region of mice (a / a) atomic exogamic, and allowed to grow until it is established a mass of approximately 100 mm 3. Epothilone B is formulated in 1% DMSO in 5% glucose in distilled water (D5W), and administered intravenously once a week for 3 weeks. Are made positive controls with clinical formulations of TAXOL® diluted 4 times with D5W and administered intravenously 3x / week, for 3 weeks, in separate doses (2 x 10 mg / kg) given with an interval of 1 hour.

The antitumor activity is expressed as% T / C (comparing Δ tumor volumes for the group of treatment with the vehicle control group) at the end of experiment. Regressions are calculated using the formula: - (T / T_ {0} -1) x 100%, where T is the tumor volume for the group of treatment at the end of the experiment, and T 0 is the volume tumor at the beginning of the experiment. Measures are taken for a period of 2 additional weeks after the fulfillment of the 3-week regular experiment, to assess the reversibility of the drug-induced body weight loss, and the sustainability of antitumor effects. The relevance is evaluated statistics using a Student's t-test of a tail, and Dunnett's evidence, or Dunn's.

Results : Table 8 summarizes the results for A549 tumors, for the standard 3-week experiment. Once-weekly administration of epothilone B produces significant relevance, the dose-dependent inhibition of tumor growth, approaching tumor stasis at the highest concentrations of the drug. Epothilone B produces marked inhibition of tumor growth at 3.5, and 3 mg / kg (T / C = 15%, and 23%, respectively). Both doses cause comparable, but reversible, body weight losses (see Table 8) of approximately 15%. Doses of 2, and 1 mg / kg produced 43%, and 74% T / C, which are statistically significant, with no increase in body weight by 2 mg / kg, and normal increase in body weight by 1 mg / kg. TAXOL®, administered 3x / week in separate doses of 2 x 10 mg / kg does not inhibit tumor growth but produces a 19% body weight loss.

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TABLE 8 Antitumor activity of epothilone B, compared with TAXOL®, in non-small cell lung tumors A549 in nude mice

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The treatments begin on the 13th after implantation (10 million cells / animal). Epothilone B is administered intravenously once a week for 3 weeks (days 13, 20, and 27) in doses of 1, 2, 3, and 3.5 mg / kg. TAXOL® is administered intravenously three times a week for 3 weeks in doses of 20 mg / kg (days 14, 17, 19, 21, 24, 26, 28, 31, and 33) as separate doses of 10 mg / kg given with an interval of one hour. The final data is recorded on day 34. A single asterisk (*) indicates p <0.05 using a Student's one-tailed test, and a double asterisk (**) indicates p <0.05 using a test of Dunnett or Dunn.

Measures are taken for an additional period of 2 weeks after the completion of the 3-week experiments, and the final data for week 5 are summarized in Table 9. The antitumor effect remains unchanged while they have recovered the body weights of the animals. T / C values for dose levels 3,5, 3, 2, and 1 mg / kg of epothilone B are 12%, 16%, 49% and 72%, respectively, and all groups have normal weight gain TAXOL® remains ineffective and the Animals show a weight gain of only 2%.

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TABLE 9 Antitumor activity of epothilone B, compared with TAXOL®, in non-small cell lung tumors A549 in naked mice (extended observation)

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The measures for the experiment are extended described in Table 1 for a period of two additional weeks. The final data is recorded on day 48. An asterisk only (*) indicates p <0.05 using a Student's one-tailed test, and a double asterisk (**) indicates p <0.05 using a test of Dunnett or Dunn.

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Example 9 Antitumor effect of epothilone B in breast tumors ZR-75-1

Table 10 shows the results of a experiment where the effect of TAXOL® and epothilone B is compared in the breast cancer cell line ZR-75-1. The methods used to This tumor model has been described previously.

From this data it is derived that (judged on the antitumor efficiency) the best dose schedule is one where Administer 4 mg / kg weekly. However, mortality is observed with all doses, suggesting that the tumor ZR-75-1 may affect health overall mice compared to other types of tumors

TABLE 10 Antitumor effect of epothilone B against human ZR-75-1 breast tumors estrogen-dependent transplanted subcutaneously in nude mice BALB / c female

26

Tumor fragments of approximately 25 mg are implanted inside the left flank of each naked mouse female (n = 6 per group); a subcutaneous estrogen pellet is placed on the opposite flank. The treatments begin on the 19th after of tumor transplantation. Epothilone B is administered in doses of 1, 2 or 4 mg / kg, intravenously once a week (days 19, 26 and 33 or every second week (days 19 and 33). Data presented are of the animals that survive on day 47, the last controls day. The antitumor activity is expressed as T / C% (average increase in animal tumor volumes treated divided by the average increase in tumor volumes of control animals multiplied by 100). Regression Tumor (%) represents the final average tumor volume compared with the average tumor volume at the beginning of treatment. The changes (Δ) in the tumor volume represent the volume tumor on the last day of treatment minus tumor volume on First day of treatment.

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Example 10 Antitumor effect of epothilone B compared to the 5-fluorouracil against colon tumors Colo 205 transplanted subcutaneously

Table 11 shows the effect of epothilone B against Colo 205 tumors transplanted subcutaneously, as well as the effect of 5-fluorouracil. Unlike treatment of HCT-15 cell line tumors, where standard treatment with 5-fluorouracil or TAXOL® treatment is not effective, here treatment with 5-fluorouracil is still effective, although much less than with epothilone B.

Together with the data from example 4 where HCT-15 cells do not respond (they are refractory) both TAXOL® and standard colon cancer treatment with 5-fluorouracil, this shows that epothilone B is certainly appropriate to treat tumors that are refractory to the known standard treatments. On the other hand, it is also more effective where standard treatments work. May deduced a preferred treatment program that is 4 mg / kg each 2 weeks (tumor regression, without dead animals). This treatment is even better than that with 5-fluorouracil where no regression is found, but only 4 of 7 animals survive.

TABLE 11 Antitumor effect of epothilone B in comparison with 5-fluorouracil against colon tumors COLO 205 human subcutaneously transplanted in nude mice BALB / c female (day 32, four days after the last treatment)

27

Tumor fragments of approximately 25 mg are implanted inside the left flank of each naked mouse female (n = 7 per group); a subcutaneous estrogen pellet is placed on the opposite flank. The treatments begin on day 14 after tumor transplantation. Epothilone B is administered at a dose of 4 mg / kg, intravenously once or once per week (days 14, 21, 28) or every second week (days 14 and 28). Be administer 5-fluorouracil intravenously in 75 mg / kg dose on days 14, 21, 28. The data presented are from the animals that survive on day 32, four days after Last treatments The antitumor activity is expressed as T / C% (average increase in animal tumor volumes treated divided by the average increase in tumor volumes of control animals multiplied by 100). Regression Tumor (%) represents the final average tumor volume compared with the average tumor volume at the beginning of treatment. The changes (Δ in tumor volumes represent volume tumor on the last day of treatment minus tumor volume on First day of treatment.

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Example 11 A phase 1 study of dose determination of an agent only epothilone B administered once weekly to patients adults with advanced solid tumors

Number of centers 2

goals

Primary: Characterize the security profile, including both acute and cumulative toxicities, and determine the maximum tolerated dose of the single agent epothilone B administered by intravenous infusion once a week to adult patients with advanced solid tumors that have failed with standard systemic therapy or for which therapy Systemic standard does not exist.

Secondary:

one.

Characterize the pharmacokinetics of the single agent epothilone B administered by intravenous infusion once every week to this patient population; the data obtained is used in conjunction with pharmacodynamic data (for example, hematological parameters), to make correlations pharmacokinetics / pharmacodynamics (PK / PD) that help predict Safety and effectiveness.

2.

Obtain preliminary activity evidence antitumor of the single agent epothilone B administered by intravenous infusion once a week to this population of patients

3.

Correlate intratumoral drug levels among adult patients with advanced solid tumors that receive single agent epothilone B by intravenous infusion a every week, with those associated with the effectiveness in preclinical models.

Four.

Gather pharmacogenetic information about tumors in tumor biopsy samples where available and accessible pre and post-therapy in order to identify genes that correlate with efficacy and response; this It is done by genetic analysis of genetic expression individual (for example, p53, Map4, and mdr1 state of expression) or well through the technology of the genetic chip.

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Design : This is a study of gradually increasing the open design dose to assess the safety, pharmacokinetics, and pharmacodynamics of epothilone B administered by intravenous infusion once a week to adult patients with advanced solid tumors that have failed therapy. systemic standard or for which there is no standard systemic therapy.

The treatment period consists of up to 24 weekly doses Patients who are prematurely discontinued They experience unacceptable toxicity or disease progression. Patients who reach a complete or partial response, or those patients with stable disease at the end of the 24 doses continue with additional treatment according to a protocol of extension at the discretion of the investigator and after approval from the sponsor. Eligible patients receive additional cycles until disease progression or unacceptable toxicity.

Phase 1 protocol design is used standard of entering 3-6 patients per cohort for set the MTD. The gradual increase in the dose comes from according to a modified Fibonacci type scheme and is based on the toxicities from the first 4 weekly doses for each patient cohort. The initial dose is 0.1 mg / m2, with doses later as follows: 0.2, 0.3, 0.5, 0.7, and 0.9 mg / m2.

Provisional BAT is defined as the level of dose immediately below that in which the toxicity Dose limit (DLT) is observed in at least two of the 3-6 patients The cohort defined as BAT provisional then admits additional patients up to a total of 12 to confirm BAT by further evaluation of the safety, pharmacokinetic, and pharmacodynamic profiles of the epothilone B.

All toxicities are defined according to the Revised Common Toxicity Criteria of the National Institute of the United States Cancer. DLTs are defined in the protocol; In general, however, the nature of a DLT is such that considered unacceptable even in the presence of a solid tumor incurable.

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Patients Inclusion criteria

The following criteria must be met for inclusion in the study:

one.

Male or female patients \ geq18 years old.

2.

Histologically documented advanced solid tumor, which has failed with standard systemic therapy and up to 1 therapy additional systemic, or for which there is no systemic therapy standard.

3.

At least one measurable site, evaluable, or not disease evaluable as defined by the Response Criteria of the Solid Tumor of the Southwestern Oncology Group (SWOG) including the marker value of the tumor that is above the limit Superior normal institutional.

Four.

Women with procreation potential should have a negative serum β-HCG pregnancy test before the start of the study drug. Sex patients male and female with procreation potential must accept the use of an effective method of birth control throughout the study and for up to 3 months after discontinuation of study drug.

5.

A rating of ≤2 in the State of Performance of the World Health Organization (WHO).

6.

Life expectancy of at least 3 months

7.

Informed written consent obtained before of any selection procedure.

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Exclusion criteria

Study exclusion is required if Any of the following points apply:

one.

Female patients who are pregnant or breastfeeding Postmenopausal women should have amenorrhea for at least 12 months to be considered without potential procreation.

2.

The patient has a medical illness not controlled and / or severe (i.e. diabetes, heart failure congestive, acute uncontrolled myocardial infarction within 6 months of study, chronic kidney disease, or no active infection controlled).

3.

The patient has a brain metastasis known.

Four.

The patient has a chronic liver disease or known acute (i.e. chronic active hepatitis, cirrhosis).

5.

The patient has a known diagnosis of infection with the human immunodeficiency virus (HIV).

6.

The patient has received any agent from investigation within 30 days prior to admission to the study.

7.

The patient received chemotherapy within 4 weeks (6 weeks for nitrosoureas or mitomycin C) prior to Admission to the study.

8.

The patient received irradiation therapy with prior to 4 weeks prior to admission to study.

9.

The patient previously received radiotherapy at ≥25% of the bone marrow.

10.

The patient had major surgery within 2 weeks before entering the study.

eleven.

The patient has a history of noncompliance with Medical regimes

12.

The patient has liver, kidney or kidney failure. hematological as defined by the following parameters of laboratory:

quad

Platelet count <100 x 10 9 / L

quad

Absolute neutrophil count (ANC) <1.5 x 109 / L

quad

ALT Serum (SGPT)> 2.5 x normal upper limit institutional (IULN)

quad

Total serum bilirubin> 1.5 x IULN

quad

Serum creatinine> 1.5 x IULN

14.

The patient has not had another primary neoplasm for the past 5 years or, in the case of non-melanomatous skin cancer and cervical carcinoma in situ , has active disease.

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Sample size : This study requires approximately 40 patients.

Treatments : Epothilone B is supplied in individual 2 ml glass ampoules formulated as 1 mg / 1 ml of clear colorless intravenous concentrate. The substance is formulated in polyethylene glycol 300 (PEG300) and diluted with 50 to 100 ml of 0.9% Sodium Chloride Injection, USP, to reach the desired final concentration of the infusion drug. It is given as a single intravenous infusion of 30 minutes every 7 days.

The initial dose level is 0.1 mg / m2. This dose is calculated as one third of the minimum toxic dose (TDL) in the most sensitive species studied which, for epothilone B, is the dog. As described above, the gradual increase in the dose proceeds according to the Fibonacci type scheme modified. The study defines delays in treatment, reductions of dose, or withdrawal of treatment for individuals who they experience hematological or other toxicity that is known to they are produced by epothilone B. Treatment continues until a maximum of 24 weekly doses unless the patient experiences disease progression or unacceptable toxicity. At the end of 24 doses, patients who have reached a complete response or partial and patients who have had a stable disease can continue with additional treatment according to a protocol extension at the discretion of the investigator and after the sponsor approval

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Security

Variables : The safety of epothilone B is assessed by physical examination and evaluation of vital signs, clinical laboratory results, adverse events, and use of concomitant medications. Adverse events are both obtained and voluntarily offered and are classified using the Revised Expanded Common Toxicity Criteria of the National Cancer Institute of the United States.

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Effectiveness

Variables : Although this phase 1 study is not designed to detect efficacy, activity is demonstrated as a function of the objective tumor response rate and the extent of progression-free and general survival. Reference tumor evaluations include the optimal evaluation of all measurable, evaluable, and non-evaluable diseases. Evaluations include physical examination and chest roentgenogram and, if appropriate, CT scan of the chest, abdomen and pelvis; sonogram of the abdomen and pelvis; bone scintigraphy, with bone roentgenogram for all known bone lesions; and determination of tumor marker values. Follow-up studies are obtained every 6 weeks and after cessation of treatment.

The objective state is evaluated clinically using the Novartis guidelines, which are based on the criteria of SWOG response. All complete and partial responses must be confirmed by a second evaluation at least four weeks later The best tumor response is calculated for each patient using the SWOG response criteria.

Pharmacokinetics : The following pharmacokinetic parameters are calculated and analyzed for cycles 1 and 2: t max, C max, λ Z, t 1/2, AUC, and R A. R_ {A} = the proportion of AUC \ tau_ {cycle} 2 / AUO \ tau_ {cycle} 1 is evaluated as an accumulation index. The preliminary assessment of dose proportionality is based on the AUC from the last dose between different dose groups. PK / PD correlations with observed toxicities (for example, hematopoietic) are performed as a safety indicator.

Pharmacodynamics : Tumor biopsy samples are obtained where it is feasible and accessible, before therapy and after the first cycle of therapy. These biopsy samples are prepared for the analysis of their genetic expressions using genetic chip technology, then analyzed separately for the p53 state, the MAP4 RNA expression, and mdr1 RNA expression.

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Statistics

Methods : Patients with adverse clinical events that emerge during treatment (especially those with dose limit toxicity) or with abnormalities in laboratory results, in vital signs, on physical examination (newly appeared or that have worsened after reference values) the values are identified and highlighted. The abnormality index is tabulated by cohort. Objective response rates (including both complete and partial responses) are presented by cohort. Descriptive statistics are used to summarize the basic pharmacokinetic parameters by cohort.

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Example 12 A specific phase 1 study on the dose of an agent single EPO906 (epothilone B) administered once every three weeks to adult patients with advanced solid tumors

Number of centers: 2

Location Glasgow, RU, & Newcastle, RU

goals

Primary: Characterize the security profile, including both acute and cumulative toxicities, and determine the maximum tolerated dose of the single agent epothilone B administered by intravenous infusion once every three weeks to adult patients with advanced solid tumors that have failed with standard systemic therapy or for which therapy Systemic standard does not exist.

Secondary:

one.

Characterize the pharmacokinetics of the single agent epothilone B administered by intravenous infusion once every three weeks to this patient population; the data obtained is used in conjunction with pharmacodynamic data to perform the pharmacokinetic / pharmacodynamic (PK / PD) correlations that They help predict safety and efficacy.

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2.

Obtain preliminary activity evidence antitumor of the single agent epothilone B administered by intravenous infusion once every three weeks to this population of patients

3.

Correlate intratumoral drug levels among adult patients with advanced solid tumors that receive single agent epothilone B by intravenous infusion a once every three weeks, with those associated with efficacy in preclinical models.

Four.

Gather information about tumors from tumor biopsy samples where available and accessible pre and post therapy in order to identify the factors biologicals that correlate with efficacy and reply.

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Design : This is a study of gradually increasing the open design dose to assess the safety, pharmacokinetics, and pharmacodynamics of epothilone B administered by intravenous infusion once every three weeks to adult patients with advanced solid tumors that have failed with standard systemic therapy or for which there is no standard systemic therapy.

The treatment period consists of up to six 21 day cycles Patients who are prematurely discontinued They experience unacceptable toxicity or disease progression. Patients who reach a complete or partial response, or those patients with stable disease at the end of the six cycles continue with additional treatment according to a protocol extension at the discretion of the investigator and after the sponsor approval Eligible patients receive cycles additional until a disease progression is observed or unacceptable toxicity.

In the absence of dose limit toxicity (DLT), the gradual increase in dose comes from the following shape:

one.

First gradual dose increase: increase in 100% dose (unless a grade 2 toxicity is identified in the first cohort, in which case the gradual increase in dose is 25% -67%).

2.

Gradual dose increases following the gradual dose increase of 100% from the first to the second cohort: gradual dose increases of 67% until Identifies grade 2 toxicity.

3.

Gradual increases in final doses following the identification of a grade 2 toxicity: increases dose increments of 25% -67%, based on the consensus reached between The researchers and the sponsor.

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The gradual increase in dose is based on the toxicities from the first cycle for each cohort of patients The provisional maximum tolerated dose (BAT) is defined as the dose level immediately below that in which the DLT is observed in at least two of the 3-6 patients The cohort defined as provisional BAT after admit additional patients up to a total of 12 to confirm BAT by further evaluation of the profiles of safety, pharmacokinetics, and pharmacodynamics of epothilone B.

Gradual dose increase will not be allowed. intrapatient

All toxicities are defined according to the Revised Common Toxicity Criteria of the National Institute of the United States Cancer. DLTs are defined in the protocol; In general, however, the nature of a DLT is such that considered unacceptable even in the presence of a solid tumor incurable.

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Patients Inclusion criteria

The following criteria must be met for inclusion in the study:

one.

Male or female patients \ geq18 years old.

2.

Histologically documented advanced solid tumor, that failed with standard systemic therapy and up to 1 additional systemic therapy, or for which there is no therapy systemic standard.

3.

At least one measurable site, evaluable, or not disease evaluable as defined by the Response Criteria of the Solid Tumor of the Southwestern Oncology Group (SWOG) including the marker value of the tumor that is above the limit Superior normal institutional.

Four.

Women with procreation potential should have a negative serum β-HCG pregnancy test before the start of the study drug. Sex patients male and female with procreation potential must accept the use of an effective method of birth control throughout the study and for up to 3 months after discontinuation of study drug.

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5.

A rating of ≤2 in the State of Performance of the World Health Organization (WHO).

6.

Life expectancy of at least 3 months

7.

Informed written consent is obtained before any selection procedure.

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Exclusion criteria

Study exclusion is required if Any of the following points apply:

one.

Female patients who are pregnant or breastfeeding Postmenopausal women should have amenorrhea due to at least 12 months to be considered without potential procreation.

2.

The patient has a medical illness not controlled and / or severe (i.e. diabetes, heart failure congestive, acute uncontrolled myocardial infarction within 6 months of study, chronic kidney disease, or no active infection controlled).

3.

The patient has a brain metastasis known.

Four.

The patient has a chronic liver disease. known or acute (i.e. chronic active hepatitis, cirrhosis).

5.

The patient has a known diagnosis of infection with the human immunodeficiency virus (HIV).

6.

The patient has received any agent from investigation within 30 days prior to admission to the study.

7.

The patient received chemotherapy within 4 weeks (6 weeks for nitrosoureas or mitomycin C) prior to Admission to the study.

8.

The patient received irradiation therapy previously within 4 weeks prior to admission to study.

9.

The patient previously received radiotherapy at ≥ 25% of the bone marrow.

10.

The patient had major surgery within 2 weeks before entering the study.

eleven.

The patient has a history of noncompliance with Medical regimes

12.

The patient has insufficient function Hepatic, renal or hepatic as defined by the following laboratory parameters:

quad

Platelet count <100 x 10 9 / L

quad

Absolute neutrophil count (ANC) <1.5 x 10 9 / L

quad

ALT (SGPT) or AST (SGOT) serum> 2.5 x limit Superior institutional normal (IULN) (> 5 x IULN for patients with liver metastasis)

quad

Total serum bilirubin> 1.5 x IULN

quad

Serum creatinine> 1.5 x IULN

13.

The patient has not had another primary malignancy during the past 5 years; however, non-melanomatous skin cancer and cervical carcinoma in situ are excluded only if the patient has active disease.

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Sample size : This study requires approximately 40 patients.

Treatments : Epothilone B is supplied in individual 2 ml glass ampoules formulated as 1 mg / 1 ml of clear colorless intravenous concentrate. The substance is formulated in polyethylene glycol 300 (PEG300) and diluted with 50 or 100 ml of 0.9% Sodium Chloride Injection, USP, to reach the desired final concentration of the infusion drug. It is given as a single intravenous infusion of 30 minutes every 21 days for six cycles.

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The initial dose level is 0.3 mg / m2. This dose is calculated as one third of the minimum toxic dose (TDL) in the most sensitive species studied which, for epothilone B, is the dog. Since there is no mortality in the minimum of the 2 doses administered to dogs in the LPG toxicology study, 0.1 mg / kg, repeated once 3 weeks later, it is estimated that the TDL is found in the range of 0.05 mg / kg. By using a factor of 20 to convert mg / kg in the dog to mg / m2 in human, this initial dose is calculated as:

1 / 3x0.05 mg / kg x 20 kg / m2 = 0.3 mg / m2

The gradual increase in dose is made of according to the scheme indicated above.

The study defines delays in treatment, dose reductions, or withdrawal from treatment for individuals experiencing hematological or other toxicity It is known that they are produced by epothilone B. The treatment continue up to a maximum of 6 cycles unless the patient experience disease progression or unacceptable toxicity. To the end of 6 cycles, patients who have reached a response complete or partial and patients who have had a disease stable can continue with additional treatment according to an extension protocol at the discretion of the investigator and after Sponsor approval.

Safety Variables : The safety of epothilone B is assessed by physical examination and evaluation of vital signs, clinical laboratory results, adverse events, and use of concomitant medications. Adverse events are both obtained and voluntarily offered and are classified using the Revised Common Toxicity Criteria of the National Cancer Institute of the United States.

Efficacy Variables : Although this phase 1 study is not designed to detect efficacy, activity is demonstrated as a function of the objective tumor response rate and the duration of progression-free and general survival. Reference tumor evaluations include the optimal evaluation of all measurable, evaluable, and non-evaluable diseases. Evaluations include physical examination and chest roentgenogram and, if applicable, CT scan of the chest, abdomen and pelvis; sonogram of the abdomen and pelvis; bone scintigraphy, with bone roentgenogram for all known bone lesions; and determination of tumor indicator values. Follow-up studies are obtained every 6 weeks and after cessation of treatment.

The objective state is evaluated clinically using the Novartis guidelines, which are based on the criteria of SWOG response. All complete and partial responses must be confirmed by a second evaluation at least four weeks later The best tumor response is calculated for each patient using the SWOG response criteria.

Pharmacokinetics : The following pharmacokinetic parameters are calculated and analyzed for cycles 1 and 2: t max, C max, λ Z, t 1/2, AUC, and R A. R_ {A} = the proportion of AUC \ tau_ {cycle} 2 / AUC \ tau_ {cycle} 1 is evaluated as an accumulation index. The preliminary assessment of dose proportionality is based on the AUC from the last dose between different dose groups. PK / PD correlations with observed toxicities (for example, hematopoietic) are performed as a safety indicator.

Pharmacodynamics : Tumor biopsy samples are obtained where pre-therapy is feasible and accessible and after the first cycle of therapy in order to identify the biological factors that correlate with efficacy and response.

Statistical Methods : Patients with adverse clinical events that emerge during treatment (especially those with dose limit toxicity) or with abnormalities in laboratory results, in vital signs, on physical examination (newly appeared or that have worsened from the reference values) are identified and the values are highlighted. The abnormality index is tabulated by cohort. Objective response rates (including both complete and partial responses) are presented by cohort. Descriptive statistics are used to summarize the basic pharmacokinetic parameters by cohort.

Discussion : Taken together, the examples provide evidence that treatment with epothilone B is effective:

to)

also against a tumor where the treatment standard fails, for example in the colon tumor where the 5-fluorouracil treatment, or where the TAXOL® treatment;

b)

also against a tumor where treatment with TAXOL® fails, for example, lung cancer, especially from non-small cells, and / or epidermoid cancer, especially cervical;

C)

also against orthotopic tumors and the formation of metastases, for example, in tumors of prostate;

d)

also against breast cancer where in vitro tests (example 3) epothilone shows greater activity than TAXOL®.

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Preferred dose regimens focus around a weekly treatment area with approximately 1/3 to 2/3 of the BAT until one treatment once with a dose of up to BAT, with a type of better treatment area that is located in the weekly administration until administration once every three weeks

Claims (34)

1. Use of epothilone B for preparation of a pharmaceutical preparation for use in the treatment of a proliferative disease where epothilone B is used in a human dose that is calculated according to the formula (I) 28 where N is the number of weeks between treatments e and is 6.

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2. The use in accordance with the claim 1 wherein epothilone B is used in a dose in human that is calculated according to formula II 29 3. The use in accordance with the claim 1 wherein epothilone B is used weekly in a dose that is between 0.1 and 6 mg / m2. 4. The use according to the claim 1 wherein epothilone B is used every third week at a dose that is between 0.3 and 18 mg / m2. 5. The use according to the claim 2 wherein epothilone B is used every third week at a dose that is between 0.3 and 7.5 mg / m2. 6. Use of epothilone B for preparation of a pharmaceutical preparation for use in the treatment of a proliferative disease where epothilone It is used in humans every week at a dose between 0.3 and 1 mg / m2. 7. The use according to the claim 1 wherein epothilone B is administered by intravenous infusion. 8. The use in accordance with the claim 1 wherein each infusion is performed for 5 to 30 minutes 9. The use according to any of claims 1 to 8 wherein the proliferative disease is refractory to treatment with one or more chemotherapeutic agents other than an epothilone, where epothilone B is administered to a human who needs such treatment at a dose that is appropriate for the treatment of said disease. 10. The use in accordance with the claim 9 wherein the refractory tumor to be treated is select from the group consisting of lung tumors, colorectal, prostate, breast or head epidermoid or neck. 11. The use according to the claim 10 wherein the tumor to be treated is a tumor colorectal that is refractory to at least one member of the class of the taxane of anticancer agents. 12. The use in accordance with the claim 11 wherein the colorectal tumor to be treated it is also refractory to at least one other chemotherapeutic agent standard. 13. The use in accordance with the claim 10 wherein the tumor to be treated is a tumor colorectal that is refractory to treatment with TAXOL and 5-fluorouracil. 14. The use in accordance with the claim 10 wherein the tumor to be treated is a tumor of prostate, and / or any metastasis of it, refractory to hormone treatment 15. The use in accordance with the claim 10 wherein the tumor to be treated is a tumor head or neck epidermoid that is refractory to treatment with at least one other chemotherapeutic agent. 16. The use in accordance with the claim 15 wherein the head and neck epidermoid tumor is refractory to treatment with TAXOL. 17. The use according to the claim 10, wherein the tumor to be treated is a tumor of lung that is refractory to treatment with at least one other chemotherapeutic agent 18. The use in accordance with the claim 17 wherein the tumor to be treated is a cancer of non-small cell lung. 19. The use in accordance with the claim 18 wherein the non-small cell lung tumor is refractory to treatment with a member of the taxane class of anti-cancer agents 20. The use in accordance with the claim 19 wherein non-small cell lung cancer It is refractory to treatment with TAXOL. 21. The use in accordance with the claim 10 wherein the tumor to be treated is a tumor of mom. 22. The use in accordance with the claim 21 wherein the breast tumor is refractory to treatment with at least one member of the taxane class of anti-cancer agents 23. The use according to the claim 10 wherein the tumor to be treated is a tumor colorectal that is refractory to standard chemotherapy. 24. The use in accordance with the claim 10 wherein the tumor to be treated is a tumor head or neck epidermoid that is refractory to treatment with at least one other chemotherapeutic agent due to resistance to multiple drugs 25. The use according to any of claims 1 to 8 wherein the proliferative disease that goes to be treated is selected from the group consisting of a tumor colorectal, a tumor of the genitourinary tract, a tumor epidermoid, a lung tumor and a breast tumor. 26. The use in accordance with the claim 25 wherein the proliferative disease that is going to be Treated is a prostate tumor. 27. The use in accordance with the claim 25 wherein the proliferative disease is a tumor head or neck epidermoid. 28. The use in accordance with the claim 25 wherein the proliferative disease is a tumor of non-small cell lung. 29. The use in accordance with the claim 25 wherein the proliferative disease is a tumor of mom. 30. The use of epothilone B for preparation of a pharmaceutical preparation; in accordance with any of claims 1 to 8 wherein the disease proliferative to be treated is a tumor resistant to multiple drugs 31. Use in accordance with any of claims 1 to 8 wherein the proliferative disease that goes to be treated is selected from the group consisting of a melanoma, an ovarian cancer, a pancreas cancer, a neuroblastoma, a head or neck cancer, a bladder cancer, a kidney cancer, brain cancer and gastric cancer. 32. The use according to any of claims 1 to 31, further comprising the step of administer (a) epothilone B in combination with (b) another agent therapeutic antitumor, being the planned combined treatment of so that component (a) and component (b) are administered to a human who needs such treatment in combination and in the amount which is therapeutically effective together against said disease proliferative 33. The use in accordance with the claim 1, wherein the proliferative disease is a tumor that is refractory to treatment with an anticancer agent of the taxane class, said tumor being selected from the group that it consists of a colorectal tumor, a prostate tumor, a tumor pancreatic and a brain tumor. 34. The use in accordance with the claim 1 wherein the proliferative disease is a carcinoma non-small cell lung resistant to multiple drugs, a multi-drug resistant breast tumor, or a tumor multi-drug resistant head and neck epidermoid.