RU2242229C2 - Applying epotilones for cancer treatment - Google Patents

Abstract

FIELD: medicine, oncology.

SUBSTANCE: invention relates to methods for treatment of cancer with different localization. Method involves administration of epotilone B with interval for from 1 to 6 weeks and a single dose is calculated as mg/m by the formula: (0.1-Y) x N wherein Y = 6; N is amount of weeks between administrations. Method provides high anti-tumor effect in combination with good tolerance of the preparation.

EFFECT: valuable medicinal properties of drugs.

28 cl, 11 tbl, 12 ex

Description

Summary of the invention

The invention relates to the treatment of proliferative disease, especially to certain treatment regimens using epothilone, especially epothilone B; preferably tumors of the gastrointestinal tract, more preferably (1) tumors of the colon and / or rectum (colorectal tumor), especially if it cannot be treated using (at least one) anti-cancer agent from the taxane class, in particular TAXOL ® (paclitaxel in the form of a dosage form intended for clinical use), and / or at least one type of standard treatment with another chemotherapeutic agent, especially 5-fluoracil; (2) genitourinary tumors, more preferably prostate tumors, including primary and metastatic tumors, especially tumors that cannot be treated with hormones ("prostate cancer that cannot be treated with hormones"), and / or treatment with other standard chemotherapeutic agents; (3) an epidermoid tumor, more preferably an epidermoid tumor of the head and neck, most preferably an oral tumor; (4) lung cancer, more preferably non-small cell lung cancer, especially any of those tumors that cannot be treated with one or more chemotherapeutic agents (primarily due to multiple drug resistance), especially treatment with anti-cancer agents of the taxane class, in particular TAXOL®; or (5) a breast tumor, more preferably a tumor that has multiple drug resistance, especially treatment with anti-cancer agents of the taxane class, in particular TAXOL®; the invention also primarily relates to the treatment of a multi-drug resistant lung tumor (preferably a small cell tumor), multi-drug resistant breast tumor or multi-drug resistant epidermoid tumor, or, more generally, the invention relates to a treatment regimen for treating the above or (in a broader sense) any other tumor, mainly immune to one or more chemo to therapeutic agents, especially those with multiple drug resistance and / or TAXOL® immune, such as melanoma, ovarian cancer, pancreatic cancer, neuroblastoma, head and neck cancer, or bladder cancer, or more generally kidney, brain cancer or stomach; comprising administering epothilone, especially epothilone B as a cytotoxic agent; the term “treatment” also includes (I) a method of treating (treating) said disease, comprising administering at least once a therapeutically effective amount of said cytotoxic agent (preferably epothilone, especially epothilone B, in each case, preferably in combination with a pharmaceutically acceptable carrier) to a warm-blooded animal, especially in need of such treatment; (Ii) the use of said cytotoxic agent for the treatment of proliferative disease; (III) the use of said cytotoxic agent for the manufacture of a pharmaceutical composition intended for the treatment of said proliferative disease (including mixing said cytotoxic agent with a pharmaceutically acceptable carrier); (IV) a pharmaceutical composition comprising a dose of said cytotoxic agent necessary to treat said proliferative disease. In a preferred embodiment, the invention relates to the treatment of patients (people) or groups of patients when other methods of treatment, especially standard treatment with chemotherapeutic agents, especially 5-fluoracil, or therapy using anti-cancer agents from the taxane class, such as TAXOL ® proved to be ineffective. The invention also relates to epothilone, in particular to epothilone B, intended for the treatment of a proliferative disease, in particular a disease that is not amenable to treatment by standard therapeutic agents.

BACKGROUND OF THE INVENTION

Cancer is the main unresolved medical problem so far. The primary treatment for the disease is often surgical treatment, radiation treatment, or a combination of these methods, but often the disease resumes (metastases appear). Chemotherapeutic treatments for most types of cancer do not lead to cure, but only slow down the development of the disease. As a rule, tumors and their metastases become insensitive to chemotherapy, a phenomenon known as the development of multiple drug resistance. In many cases of a tumor, from the very beginning, resistance to certain classes of chemotherapeutic agents is inherent [see DeVita V.T., Principles of Cancer Management: Chemotherapy. B: Cancer. Principles and Practice of Oncology. DeVita V.T. et al. (ed.), 5th ed., Lippincot-Raven, Philadelphia, New York, pp. 333-347, (1977); or Cleton F.J., Chemotherapy: general aspects. B: Oxford Textbook of Oncology; Peckham M. et al., Oxford University Press, Oxford, New York, Tokyo, vol. 1, pp. 445-453 (1995)]. This is the case, for example, in the case of a lung tumor, especially small cell lung carcinoma, or also epidermoid tumors, such as epidermoid tumors of the head and neck, especially the mouth, or also breast tumors. Other mechanisms leading to the fact that tumors cannot be cured (cannot be treated) can be, for example, the presence of tubulin mutations or mechanisms mediated by glutathione.

353-374 (1997)]. Также и для рака мочеполовой системы, прежде всего рака предстательной железы, представляющего собой еще одну нерешенную медицинскую проблему, первоочередное лечение, как отмечалось выше для колоректального рака, сталкивается с аналогичными проблемами. Основное химиотерапевтическое лечение при рецидиве рака предстательной железы включает применение антиандрогенов и рецидив часто является зависимым от андрогена. Однако такое лечение обеспечивает только замедление развития болезни, поскольку опухоли почти всегда становятся невосприимчивыми к антиандрогенам в течение периода времени от 6 месяцев до 2 лет (опухоли предстательной железы, не поддающиеся лечению гормонами). Химиотерапия болезней на этой стадии развития устойчивости к антиандрогенам включает применение митоксантрона или других классических противораковых цитотоксических агентов, однако все они считаются недостаточно эффективными [см. Oesterling и др., Cancer of the prostate. B: Cancer. Principles and Practice of Oncology; DeVita и др. (ред.), 5-е издание, Lippincott Raven, Philadelphia, New York, стр. 1322-1386, 1997; Sternberg, Cancers of the genitourinary tract. B: Cavalii и др. (ред.). Textbook of Medical Oncology; или Roth B.J., Semin. Oncol.

(6 дополн. 14), 49-55 (1996)].Colorectal cancer, primarily colorectal cancer, is a special case of unresolved medical problems in the treatment of cancer. The primary treatment for the disease is often surgical treatment, radiation treatment, or a combination of these methods, but often the disease resumes (metastases appear). Major chemotherapeutic treatments for relapse of colorectal cancer include the use of 5-fluoracil. However, in the best case, such treatment provides only a slowdown in the development of the disease, since tumors usually become immune to treatment. Chemotherapy at this stage of the development of disease resistance involves the use of other classical cytotoxic agents, but all of them are considered insufficiently effective [see Cohen et al., Cancer of the colon. B: Cancer. Principles and Practice of Oncology; DeVita et al. (Ed.), 5th edition, Lippincott Raven, Philadelphia, New York, pp. 1144-1197, 1997; or Rowinsky, Ann. Rev. Med.

353-374 (1997)]. Also for cancer of the genitourinary system, especially prostate cancer, which is another unresolved medical problem, the primary treatment, as noted above for colorectal cancer, faces similar problems. The main chemotherapeutic treatment for relapse of prostate cancer involves the use of antiandrogens and the relapse is often androgen dependent. However, such treatment only provides a delay in the development of the disease, since tumors almost always become immune to antiandrogens over a period of time from 6 months to 2 years (prostate tumors that cannot be treated with hormones). Chemotherapy of diseases at this stage in the development of resistance to antiandrogens involves the use of mitoxantrone or other classical anti-cancer cytotoxic agents, but all of them are considered insufficiently effective [see Oesterling et al., Cancer of the prostate. B: Cancer. Principles and Practice of Oncology; DeVita et al. (Eds.), 5th edition, Lippincott Raven, Philadelphia, New York, pp. 1322-1386, 1997; Sternberg, Cancers of the genitourinary tract. B: Cavalii et al. (Eds.). Textbook of Medical Oncology; or Roth BJ, Semin. Oncol.

(6 appendix 14), 49-55 (1996)].

353-374 (1997)].Among the cytotoxic agents intended for the treatment of tumors, TAXOL®, a microtubule stabilizing agent in great demand on the market, is a very important compound [see Rowinsky EK, The development and clinical utility of the taxane class of antimicrotubule chemotherapy agents; Ann. Rev. Med.

353-374 (1997)].

However, TAXOL® has many disadvantages. First of all, a serious problem is its poor solubility in water. It turns out to be necessary to administer TAXOL® as a composition with Cremophor EL® (polyethoxylated castor oil; BASF, Ludwigshafen, Germany), which has serious side effects, causing, among other things, allergic reactions, which, in one case, were reported to have even led to the death of the patient . Moreover, it is known that some types of tumors are immune to TAXOL® treatment even if the drug is administered as an initial (intensive) therapy or the tumors develop resistance to TAXOL® after several cycles of use.

196-208 (1995); или Arbuck S.G. и др., J. Natl. Cancer Inst. Monogr.

11-24 (1993)]. Например, эффективность TAXOL® может быть серьезно ограничена механизмами приобретенной устойчивости к лекарству, опосредованных различными механизмами, такими как сверхэкспрессия фосфогликопротеинов, действующих как насосы, откачивающие лекарства.Although the taxane, which is a member of the class of anti-cancer anti-microtubular agents, has been named "probably the most important contribution to the arsenal of chemotherapeutic agents for the fight against cancer over the past few decades" [see Rowinsky EK, Ann. Rev. Med. 48, 353-374 (1997)] and despite the commercial success of TAXOL®, there are limitations to the effective use of TAXOL®. Treatment with TAXOL® is associated with a number of significant side effects and some important classes of solid tumors, namely, colon and prostate tumors that are poorly affected by this compound (see Rowinsky EK, in the aforementioned place). In particular, it is believed that when used alone, TAXOL® has weak clinical activity against colorectal cancer, cancer of the kidney, prostate, pancreatic gland, stomach and brain [see Rowinsky EK, in the mentioned place; Bitton RJ et al. Drug Saf.

196-208 (1995); or Arbuck SG et al., J. Natl. Cancer Inst. Monogr.

11-24 (1993)]. For example, the efficacy of TAXOL® can be severely limited by mechanisms of acquired drug resistance mediated by various mechanisms, such as overexpression of phosphoglycoproteins acting as drug pumping pumps.

Thus, there is a need to find compounds and develop appropriate drug regimens using these compounds that expand the arsenal of drugs for treating cancer, especially in most cases when treatment with taxanes and other compounds with anticancer activity does not ensure survival during long period.

560-563 (1966); или у Hoefle и др. DE 4138042), описываются формулой:Epothilones, primarily epothilones A and B, represent a new class of microtubule stabilizing cytotoxic agents (see Gerth, K. et al., J. Antibiot.

560-563 (1966); or in Hoefle et al. DE 4138042) are described by the formula:

where R represents a hydrogen atom (epothilone A) or methyl (epothilone B).

These compounds have the following advantages:

(I) they have better solubility in water than TAXOL®, and are therefore more suitable for inclusion in the composition; and

2325-2333 (1995); и у Bollag D.M., Exp. Opin. Invest. Drugs

867-873 (1997)]; и(II) there is evidence that in experiments using cell cultures, they also have activity in relation to cell proliferation, which, as a result of the activity of the “pumping pump” associated with P-glycoprotein, have become more resistant to drugs, showing resistance to treatment with other chemotherapeutic agents, for example TAXOL® [see by Bollag DM et al., "Epotilones, a new class of tubule-stabilizing agents with a Taxol-like mechanism of action". Cancer research

2325-2333 (1995); and Bollag DM, Exp. Opin. Invest. Drugs

867-873 (1997)]; and

(III) it was established that despite the fact that they apparently have the same target, i.e. a sterically proximal microtube binding site, epothilones have a higher activity against the resistant to TAXOL® line of ovarian carcinoma cell lines with modified β-tubulin [see at Kowalski R.J. et al., J. Biol. Chem. 272 (4), 2534-2541 (1997)].

9642-9647 (1998)]. При создании изобретения неожиданно было установлено, что на самом деле могут быть найдены схемы приема лекарственных средств, которые позволяют, с одной стороны, осуществлять лечение опухолей с помощью эпотилонов, прежде всего эпотилона В; и с другой стороны, проводить лечение определенных групп пациентов, которые являются невосприимчивыми к другим видам лечения вследствие множественной устойчивости к лекарствам, таким как таксаны, например TAXOL®, не поддаются лечению вследствие развития множественной устойчивости к лекарствам и/или какого-либо другого механизма.On the other hand, they are highly toxic and therefore it was believed that their use for the treatment of cancer in vivo is practically impossible [see, for example, PNAS

9642-9647 (1998)]. When creating the invention, it was unexpectedly found that, in fact, drug regimens can be found that allow, on the one hand, the treatment of tumors with epothilones, especially epothilone B; and on the other hand, to treat certain groups of patients who are immune to other types of treatment due to multiple drug resistance, such as taxanes, for example TAXOL®, are not treatable due to the development of multiple drug resistance and / or some other mechanism.

It is an object of the present invention, first of all, to develop schemes for effective in vivo treatment using epothilones, preferably epothilone A or especially epothilone B, for treating tumors related diseases, for example melanoma, ovarian cancer, pancreatic cancer, neuroblastoma, head and neck cancer , cancer of the bladder, kidney, brain, stomach, or preferably colorectal cancer, cancer of the prostate, breast, lung (especially non-small cell lung cancer) epidermoid, e.g. epidermoid carcinoma of the head and neck, especially mouth.

Although in general the treatment regimen allows the treatment of various types of tumors already in the form of the main treatment, the invention preferably relates to the treatment of tumors that are assumed or established to be resistant to treatment with other chemotherapeutic agents, for example, standard treatment with one or more other chemotherapeutic agents, before total fluoracil and / or taxane, for example, treatment using TAXOL®.

When creating the invention, it was unexpectedly found that the proliferation of even such tumor cells and tumors that are not amenable to standard treatment with other chemotherapeutic agents, for example, 5-fluoracil; and / or treatment with a compound from the class of taxanes, most preferably TAXOL®, especially tumors such as a colorectal tumor, especially one that also does not respond to standard treatment, for example, with 3-fluoracil; or a lung tumor, especially non-small cell lung cancer; an epidermoid tumor, more preferably an epidermoid tumor of the head and neck, such as an oral tumor; or a breast tumor; and / or their metastases can be reduced or suspended, so that even regression or disappearance of the tumor is possible.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

The present invention preferably relates to the following subject, which is part of the invention:

If the term “proliferative disease treatment” or tumor, cancer or the like is used in the description, then

a) a method of treatment (the same as treatment) of a proliferative disease, comprising the step of administering (at least in a single treatment) an epothilone, preferably epothilone A and / or B, especially epothilone B, (preferably in combination with a pharmaceutically acceptable carrier ) to a warm-blooded animal, especially a person in need of such treatment, in a dose that allows the treatment of the disease (the same as a therapeutically effective amount), preferably in a dose (in the amount) indicated (m) above and below in the infusion the present description as preferred (s);

b) the use of epothilone, preferably epothilone A and / or B, especially epothilone B for the treatment of proliferative disease, or epothilone, especially epothilone B, for use in the treatment of the disease (especially in humans);

c) the use of epothilone, preferably epothilone A and / or B, in particular epothilone B, for the preparation of a pharmaceutical composition for the treatment of proliferative disease; and / or

g) a pharmaceutical composition comprising a dose of epothilone, preferably epothilone A and / or B, especially epothilone B, suitable for the treatment of proliferative disease; or any combination of the objects of the invention indicated in sections a), b), c) and d) that is valid for patenting in the country in which this application is filed;

d) a method of using epothilone for the preparation of a pharmaceutical composition intended for the treatment of proliferative disease, comprising mixing epothilone with a pharmaceutically acceptable carrier. In cases where instead of the term “proliferative disease”, the term tumor-related disease or a specific tumor is used (for example, a colon tumor, colon carcinoma or colon cancer; or a prostate tumor, prostate carcinoma or prostate cancer) , it should be understood that it applies to all the objects listed in sections a) -e), this means that in accordance with the patented object of the invention in sections a) -e) instead of the concept of "proliferative disease" It is recorded corresponding to a disease associated with a tumor; preferably any treatment according to a) to e) relates to the treatment of people.

The first object of the present invention relates to the treatment of a proliferative disease that cannot be treated with one or more other chemotherapeutic agents, in which epothilone, preferably epothilone A and / or B, especially epothilone B, is administered to a warm-blooded animal, especially a person, preferably a person in need in such treatment, especially in a therapeutically effective amount.

The second object of the present invention relates to a treatment regimen for in vivo proliferative disease, in particular cancer, which cannot be treated with one or more other chemotherapeutic agents, especially from the taxane class, such as TAXOL®, and / or 5-fluoracil, according to which epothilone, preferably epothilone A and / or B, especially epothilone B, is administered to a warm-blooded animal, especially a human, at a dose in the range of from about 1 to about 100%, preferably from about 25 to about 100% (single administration) of the maximum tolerated dose (MTD); and one or more (preferably from two to seven) additional doses, each of which is preferably in the above dose range, is administered during one or more additional treatment cycle (s), primarily with an interval between treatment cycles of one week or more than one week after the previous treatment, more preferably after about 1 to about b weeks, most preferably about 1 to about 3 weeks after the previous treatment cycle, respectively etstvenno. In general, such a treatment regimen in which a high dose is administered over two or more treatment cycles with interruptions of one to six, preferably within one to three weeks between administrations, is preferable to more frequent treatments with lower doses before all because it allows to reduce the frequency and duration of hospitalization and because such a regimen causes stronger antitumor effects and has less toxicity compared to more frequent abotkami and stronger antitumor efficacy compared to less frequent treatments.

Preferably, the dose of epothilone B to be administered to humans is calculated according to formula (I)

where N (above and below in the present description is an integer (for example, 1, 2 or 3) or a fractional number (for example, 1.5 or 2.3)) denotes the number of weeks between treatments (preferably a number in the range from approximately 1 to about 6 (this corresponds to an interval of from about 1 to about 6 weeks), especially from about 1 to about 3 (this corresponds to a preferred interval of about 1 to about 3 weeks) and y is 6 or preferably 5, more preferably 4.

More preferably, the dose for treatment is calculated according to formula II

even more preferably, the dose for treatment is calculated according to formula III

and most preferably, the dose for treatment is calculated according to formula IV

where in each of formulas II-IV, N has the value indicated for formula I. For doses calculated according to any of formulas I-IV, the following condition must be met: the dose for a single administration, even if the calculation gives a higher value, should not exceed approximately 18 mg / m ² . Preferably, a dose of from about 0.1 to about 6 mg / m ² , preferably from about 0.1 to about 5 mg / m ² , more preferably from about 0.1 to about 3 mg / m ² , even more is used for weekly treatment preferably from 0.1 to 1.7 mg / m ² and most preferably from about 0.3 to about 1 mg / m ² ; for a three-week treatment regimen (treatment every three weeks or every third week), a dose of from about 0.3 to about 18 mg / m ² , preferably from about 0.3 to about 15 mg / m ² , more preferably from about 0 3 to about 12 mg / m ² , even more preferably from about 0.3 to about 7.5 mg / m ² , even more preferably from about 0.3 to about 5 mg / m ^2, and most preferably from about 1, 0 to about 3.0 mg / m ² . This dose is preferably administered to a person intravenously (iv) for 2-180 minutes, preferably for 2-120 minutes, more preferably for a period of time from about 5 to about 30 minutes, most preferably for a period of time from about 10 up to about 30 minutes, for example for about 30 minutes

Preferably, especially in the case of weekly treatment, in order to allow sufficient recovery after the previous treatment, for example after 3, 4, 6, 8 or more treatment cycles, rest periods of more than one week should be arranged, more preferably for 2-10 weeks, even more preferably within 3-6 weeks, depending on the condition of the patient.

A third object of the invention relates to a treatment regimen for an in vivo proliferative disease, especially one that cannot be treated with one or more other chemotherapeutic agents, according to which epothilone, preferably epothilone A and / or B, especially epothilone B, is administered weekly to a warm-blooded animal, especially a human , in a dose of less than 80%, more preferably less than 50% of the maximum tolerated dose (MTD).

A fourth aspect of the invention relates to the treatment of an in vivo proliferative disease, especially one that cannot be treated with one or more other chemotherapeutic agents, in particular 5-fluoracil or a microtubule stabilizing agent from the taxane class, especially TAXOL®, for example, which has multidrug resistance to tumors according to which a warm-blooded animal, first of all, a person is administered epothilone, first of all epothilone B.

A fifth aspect of the invention relates to the treatment of an in vivo proliferative disease, especially one that cannot be treated with one or more other chemotherapeutic agents, by coadministration of (a) epothilone, preferably epothilone A and / or epothilone B, especially epothilone B, in combination with (b) another antitumor chemotherapeutic agent, preferably the regimen of joint treatment is such that components (a) and (b) are administered to a warm-blooded animal, especially to a person (primarily in need of such treatment) in combination in amounts that are therapeutically effective against a proliferative disease, preferably one that can be treated by administration of epothilone, more preferably epothilone A and / or epothilone B, especially epothilone B; preferably, such administration is carried out to a person who suffers from a tumor that cannot be treated with other chemotherapeutic agents, for example, first of all with treatment with 5-fluoracil or especially with anti-cancer agents from the taxane class, such as TAXOL®.

In this regard, the invention also relates to a combination composition comprising components (a) and (b) as defined in the last paragraph.

The invention also relates to a product for the treatment of proliferative disease, which includes component (a) and component (b), as defined above in the second of this paragraph, in combination with one or more pharmaceutically acceptable carriers or without them, which is a combined composition intended for simultaneous or phased administration to a warm-blooded animal, especially a person, for a sufficiently short period of time so that the active substances ulation other as component (a) and component (b), we could share enhance antiproliferative activity (especially against proliferating cells) in the body of a warm-blooded animal.

The general concepts used above and below in the present description, unless otherwise indicated, have the following meanings:

Proliferative disease is mainly a tumor-related disease (or cancer) (and / or any metastases), regardless of where the tumor or metastases are located, more preferably a tumor selected from the group consisting of breast cancer, cancer of the genitourinary system, lung cancer, cancer of the gastrointestinal system, epidermoid cancer, melanoma, ovarian cancer, cancer of the pancreatic gland, neuroblastoma, cancer of the head and neck (if you use this concept, it means cancer of the head and / or neck, but this does not mean only head and neck cancer, but also head or neck cancer) or bladder cancer, or more broadly, kidney, brain or stomach cancer; more preferably (I) a tumor selected from the group comprising a breast tumor; an epidermoid tumor, especially an epidermoid tumor of both the head and neck, preferably the mouth; and a lung tumor, especially a non-small cell lung tumor; or from the group comprising a tumor of the gastrointestinal system, especially a colorectal tumor; and a tumor of the genitourinary system, especially a tumor of the prostate gland (primarily a tumor of the prostate that cannot be treated with hormones); or (II) (more preferably) a proliferative disease that cannot be treated with other chemotherapeutic agents, especially the corresponding tumor (and / or any metastases), more preferably a tumor selected from the group consisting of tumors that are not amenable to standard treatment with other (and) chemotherapeutic (i) with agent (s), primarily with 5-fluoracil and / or (preferably) a microtubule stabilizing agent from the taxane class, most preferably TAXOL®, even more preferably a tumor selected from the group, incl. a swelling tumor of the gastrointestinal system, for example, the colorectal system (primarily not amenable to standard treatment, for example, using 5-fluoracil and / or TAXOL®); and the genitourinary system, for example, the prostate gland (and / or its metastases, especially its metastases); most preferably a tumor of the gastrointestinal system, especially colorectal cancer; or (III) a tumor that cannot be treated with other chemotherapeutic agents due to the development of multiple drug resistance, especially not treatable with microtubule stabilizing agents from the taxane class, preferably TAXOL®, most preferably resistant to many drugs, especially TAXOL® a lung tumor (especially a non-small cell lung tumor), a multi-drug resistant breast tumor, or a multi-drug resistant epidermoid tumor, prefer a particularly epidermoid tumor of the head and neck, most preferably an oral tumor.

In a broader sense, the invention relates to a proliferative disease, which can be selected from hyperproliferative conditions such as hyperplasia, fibrosis (primarily lung fibrosis, but also other types of fibrosis, such as kidney fibrosis), angiogenesis, psoriasis, atherosclerosis, and smooth muscle proliferation tissue in the blood vessels, such as angioplasty caused by stenosis or restenosis.

If a tumor, a disease associated with a tumor, carcinoma or cancer is mentioned above and below in the present description, it also implies metastases in the original organ or tissue, and / or in any other place, individually or in combination, regardless of where they are located swelling and / or metastases.

The term “non-treatable” means that with an appropriate proliferative disease (especially in the case of a tumor and / or any of its metastases) in the case of treatment with (at least one) chemotherapeutic agent other than epothilone, no antiproliferative is observed after treatment with such an agent reaction or it manifests itself only to a small extent (there is no inhibition of tumor growth, or it occurs only to a small extent), this means that the tumor in a warm-blooded animal, especially in humans eka, cannot be cured at all, or the results turn out to be unsatisfactory when using other (preferably standard) chemotherapeutic agents (preferably those indicated above, especially 5-fluoracil (especially in the case of colorectal cancer, such as colon cancer), antiandrogens, or preferably mitoxantrone (especially in the case of prostate cancer), or antiestrogens such as letrozole (preferably in the case of breast cancer); or preferably taxane-class chemotherapeutic agents, such as, for example, TAXOTERE® or TAXOL®; for example, tumor growth does not stop, it only slows down slightly or its regression is not observed. If in the present description the treatment of non-treatable tumors and the like is mentioned, it should be understood that this applies not only to the tumor (s) for which the use of one or more chemotherapeutic agents has been unsuccessful in treating the patient, but also to the tumor (s), for which (s) the fact that it (s) is not amenable to treatment is detected by other means, for example, by biopsy and cultivation in the presence of chemotherapeutic agents.

If the term “not treatable with TAXOL®” is used above and below in the present description, it should be understood that this concept applies not only to the final product, but also to paclitaxel, the active substance TAXOL®. “Hormone-free” or “hormone-free” in the case of a tumor of the genitourinary system, especially a tumor of the prostate gland, means that it cannot be treated with antiandrogens.

TAXOL® preferably refers to a final product containing paclitaxel, but in a broad sense this name also includes paclitaxel per se, which is part of any other paclitaxel-based composition in combination with one or more carriers (s).

Preferably, the term “non-treatable” means that using a standard dose, for example, by in vivo or in vitro measurements, a decrease in tumor growth of less than 50% is obtained (i.e., T / C% is equal to or greater than 50%), compared with control without the use of chemotherapeutic agents.

A multi-drug resistant tumor-related disease is a disease for which resistance to one or more chemotherapeutic agents has been found, including members of the taxane class, especially TAXOL®, or members of the anthracycline class, especially ADRIAMYCIN®. The basis of this resistance is export, due to the action of an energy-dependent pump (primarily ATP) located on the surface of the cells of the corresponding tumor, primarily associated with a member of the P-glycoprotein family, especially P-glycoprotein (P-gp) itself. In the present invention, it is understood that other mechanisms alone or in combination with the above mechanism may cause the tumor to become untreatable with chemotherapeutic agents other than epothilone. Such stability can be caused, for example, by changes in the target of the drug (in the case under consideration, primarily in microtubes), changes in intracellular metabolism, which can inactivate the compound, or changes in the physiology of the cell, which can facilitate switching off or neutralizing the mechanism of action of the drug.

In the context of the present description, the term "another chemotherapeutic agent" or "standard chemotherapeutic agent" refers primarily to any chemotherapeutic agent other than epothilone; preferably one indicated in the introduction, especially 5-fluoracil (especially in the case of colorectal cancer, for example colon cancer), an antiandrogen or mitoxantrone (especially in the case of prostate cancer), or an anti-estrogen such as letrozole (especially in case of breast cancer); in particular, the term refers to 5-fluoracil or (more preferably) microtubule stabilizing agents from the taxane class, such as preferably TAXOTERE® or more preferably TAXOL®. “Standard treatment with other chemotherapeutic agents”, “other chemotherapeutic treatment” or “standard chemotherapy” refers to treatment with at least one such “other” or “standard chemotherapeutic agent”.

The term epothilone refers to any epothilone or epothilone derivative. Preferably, the term “epothilone” means epothilone A, epothilone B, any derivative of epothilone described in WO 98/25929 (incorporated herein by reference), or any mixture thereof; more preferably, it refers to epothilone A and / or epothilone B and most preferably it refers to epothilone B.

In all cases mentioned above and below in the present description, the administration can be carried out orally, however, from the point of view of achieving better and better controlled bioavailability, it is more preferable to administer it parenterally, especially intravenously, for example by infusion or injection. Further, if the term “infusion” is used, it preferably means intravenous infusion, which is the most preferred route of administration.

Further, the basis for illustration are data related to adult patients. However, it is taken for granted that the present invention also relates to the treatment of proliferative diseases in pediatrics. For this, the doses should be adjusted according to standard methods and taking into account the age, condition and other individual characteristics of the patient.

The maximum tolerated dose (MTD) is determined according to standard methods; preferably, for warm-blooded animals, MTD in the case of oral or intravenous administration is defined as the dose for a single administration at which death does not occur and the weight loss of the body is less than 40, preferably less than 25%, in the treated warm-blooded animal (this concept mainly refers to animal; for humans, see below).

(15), стр. 1138-1147, 1997.MTD can vary depending on the group of patients, which can be determined by the type of tumor, age, gender, stage of tumor development, etc. If for animals the most preferred way to determine MTD can be similar to that described in the examples below, for people MTD is usually determined starting with a single administration of a very low dose, for example 1/10 LD ₁₀ (i.e., the dose causing death 10 % of animals) for the most sensitive species of animals for which toxicological studies were carried out, for example for epothilones (primarily for epothilone B), such a dose is in the range from 0.1 to 25 mg / m ² , for epothilone B, preferably in the range from 0, 1 to 2.5 mg / m ² most preferably in the range of 0.1 to 0.33 mg / m ² . The dose increase for the next dose level is 100% until a degree 2 toxicity is discovered according to the revised General Cancer Institute Revised Common Toxicity Criteria, in which case the dose increase is 67%. Increasing the dose to obtain subsequent dose levels is in the range of 25 to 67%. For example, three patients are usually treated at the same dose level and observed for signs of acute toxicity during one course of treatment until other patients are involved in the experiment. If none of the three patients shows signs of CCT (dose limiting toxicity), then the next group of three patients is treated with the next higher dose. If two or three patients from the group show signs of CCT, then another three patients are treated with the next lower dose until six patients are treated with the same dose. If one of the three dose-treated patients shows signs of CCT, then another three patients are treated with a dose of the same level. If among these six patients there is one case of CCT, then the next group is treated with a higher dose. In general, if among six patients treated with the same dose, two or more showed signs of CCT, it is considered that the MTD is exceeded, and the next three patients are treated, as described above, with the next lower dose. For MTD, the highest dose is taken, for which the number of cases of CCT is less than 33%. As a rule, consecutive courses of increasing doses for the same patient, i.e. increasing doses in the body of one patient. Alternatively, sequential doses can be determined using modified Fibonacci series, in which case dose increments to obtain a dose of the next level higher than the initial dose are 100, 67, 50 and 40%, and then 33% for all subsequent dose levels. Finally, MTD can be determined according to the methods described by Simon R. et al., J. Nat. Cancer Inst.

(15), pp. 1138-1147, 1997.

CCT usually includes any drug-related death and most drug-related toxicities of grades 3 and 4, including but not limited to febrile neutropenia (see also the general toxicity criteria of the US National Cancer Institute, see above for examples). For humans, preferred doses for treatment are determined according to the above formula I, more preferably according to formula II, most preferably according to formula III (provided that none of the single doses exceeds 18 mg / mm ² ).

Preferably, a dose of from about 0.1 to about 6 mg / m ² , preferably from about 0.1 to about 5 mg / m ² , more preferably from about 0.1 to about 3 mg / m ² , even more is used for weekly treatment preferably from 0.1 to about 1.7 mg / m ² and most preferably from about 0.3 to about 1 mg / m ² ; for a three-week treatment regimen (treatment every three weeks or every third week), a dose of from about 0.3 to about 18 mg / m ² , preferably from about 0.3 to about 15 mg / m ² , more preferably from about 0 3 to about 12 mg / m ² , even more preferably from about 0.3 to about 7.5 mg / m ² , even more preferably from about 0.3 to about 5 mg / m ^2, and most preferably from about 1, 0 to about 3.0 mg / m ² . This dose is preferably administered to a person intravenously (iv) for 2-180 minutes, preferably for 2-120 minutes, more preferably for a period of time from about 5 to about 30 minutes, most preferably for a period of time from about 10 up to about 30 minutes, for example for about 30 minutes

When using the results obtained in experiments on animals, doses suitable for humans (adults) can be roughly determined as follows:

A dose of 1 mg / kg for a mouse corresponds to a dose of 3 mg / m ² for a human.

A sufficient recovery from the previous treatment in relation to warm-blooded animals is understood primarily to restore the body weight of the treated individual to the initial level determined before the first dose, preferably at least 95% of this weight. For humans, recovery from each previous dose administered is preferably defined as recovery from any condition characterized by a toxicity of degree 3 or 4, including, for example, reaching a platelet level of at least 100,000 / mm ³ and a neutrophil level of at least 1,500 cells / mm ³ whole blood.

Treatment can be repeated if after the first treatment the desired result is not achieved until progressive development of the tumor is detected or until other causes (for example, the patient’s condition) require treatment to be stopped. For humans, treatment with doses ranging from about 25% to about 100% of the MTD is preferably repeated every 1-10, especially 2-10 weeks, preferably every 1-10 weeks or every 3-6 weeks, until progressive development of the disease will be detected, an unacceptable level of toxicity, until 1 or preferably 2 cycles are carried out after a complete reaction is achieved, or if the patient refuses for some reason.

Preferably, in the case of weekly human treatment with epothilone, the dose is in the range of from about 5 to about 60%, preferably from about 10 to about 60%, for example from about 5 to about 35% of the MTD, especially in the range of from about 30 to about 35% from MTD. Preferably, every three weeks, the dose of epothilone B is in the range of from about 5 to about 60%, preferably from about 10 to about 60%, even more preferably from about 10 to about 45%, most preferably from about 30 to about 45%, MTD.

Most preferably, treatment is stopped after 3-8, especially 3-5 weekly administrations, followed by a rest period of 2-5, for example, 2 weeks, before resuming treatment consisting of once or again weekly or every two weeks administered. In particular, in the case of weekly treatment with epothilone B, it is stopped after 3-8 administrations, followed by a rest period of 2-4, for example, 2 weeks, before resuming treatment by weekly administration.

The introduction of component (a), which is epothilone (s) A and / or B, especially B, is preferably carried out as described above, primarily using the above specific treatment regimens.

The introduction of component (b) is preferably carried out in accordance with the treatment regimen known to the person skilled in the art.

In a first preferred embodiment of the invention, component (b) is administered prior to administration of component (a), preferably according to a treatment regimen comprising one or more administrations of component (b) prior to treatment with component (a), preferably so that treatment with component (b) ended at least two, preferably 5-10, for example, approximately 5 days before the start of treatment with component (a), which is then administered one or more times, preferably one to five, most preferably one or two times.

In a more preferred embodiment, component (a) is administered according to a treatment regimen at 3-week intervals prior to administration of component (b), preferably according to a treatment regimen comprising one administration of component (a) prior to treatment with component (b), more preferably treatment with component (a) ends immediately before treatment with component (b), which is administered after it.

In a second more preferred embodiment, component (a) is administered according to a weekly treatment regimen. On the other hand, component (b) is administered according to the treatment regimen at 3-week intervals, with each administration being carried out immediately after the completion of every third administration of component (a).

In a third more preferred embodiment of the invention, component (a) is administered according to a weekly treatment regimen prior to treatment with component (b), preferably according to a treatment regimen comprising one administration of component (a) prior to treatment with component (b), more preferably such treatment with component ( a) ends immediately before treatment begins with component (b), which is administered after it.

In the context of the present description, the term "other chemotherapeutic agent" means primarily any chemotherapeutic agent that is used or can be used to treat tumor-related diseases, including those chemotherapeutic agents that are derivatives of agents from the following classes:

(A) alkylating agents, preferably crosslinked chemotherapeutic agents, preferably bisalkylating agents;

(B) anti-cancer antibiotics, preferably doxorubicin (ADRIAMYCIN®, RUBEX®);

(B) antimetabolites;

(D) plant alkaloids;

(D) hormonal agents and antagonists;

(E) biological response modifiers, preferably lymphokines or interferons;

(G) protein inhibitors such as tyrosine kinases and / or serine / threonine kinases;

(H) antisense oligonucleotides or oligonucleotide derivatives;

or

(Ii) a variety of agents or other agents with a different or unknown mechanism of action.

The term "therapeutically effective in combination therapy for a proliferative disease that is treatable by administering epothilone A and / or epothilone B, especially epothilone B" preferably means that in the case of the proliferative disease mentioned above, especially a tumor-related disease, preferably a reaction manifests itself in reducing proliferation, for example, in reducing tumor growth or even (more preferably) in regressing the tumor, or (most preferably) in the disappearance of the tumor ("p lnaya reaction ").

Preferably, the term “amount therapeutically effective in co-therapy for a proliferative disease that is treatable by administration of epothilone A and / or epothilone B, especially epothilone B” means any number of components (a) and (b) included in the composition, which when used together, it causes a decrease in the proliferation of cells causing any of these proliferative diseases, especially tumor cells (including metastases) (primarily a decrease in tumor growth), or preferably even causes regression, more preferably even a partial or complete disappearance of such cells (first of all, tumor regression, preferably a complete reaction, meaning the disappearance of the tumor (s)). This concept includes not only combinations of any components (a) and (b) in which doses (a) and (b) are effective for proliferation already when each of the components is used separately, but also such doses of any such components, in which the use of the components individually does not have an effect or has only a weak effect, however, when used together, it causes pronounced antiproliferative effects, i.e. reduces proliferation or preferably causes regression of proliferating cells, or even leads to a cure for proliferative disease. In addition, in the present description, the term “combination” is used not only to mean fixed combinations of components, but also any combination of components (a) and (b), intended for simultaneous or sequential use for a sufficiently short period of time, such that substances of both component (a) and component (b) could mutually enhance antiproliferative activity, for example, in the patient’s body.

In the context of the present description, the term "combined composition comprising component (a) and component (b)" means any combination representing a set of elements or a separate combined composition comprising components (a) and (b) in the form of a pharmaceutical product, which preferably a pharmaceutically acceptable carrier is present. Preferred carriers are set forth in the Pharmaceutical Compositions Section below.

In the context of the present description, the term "product comprising component (a) and component (b)" preferably means a product comprising (a) at least one compound selected from epothilone A and (preferably) epothilone B and

(b) at least one other chemotherapeutic agent, in which one or more pharmaceutically acceptable carriers are present or may be absent, in the form of a combined composition intended for simultaneous or sequential use, preferably for a sufficiently short period of time, such that the active substances as component (a) and component (b) could mutually enhance antiproliferative activity against proliferating cells, especially in the patient’s body, For the treatment of a proliferative disease that can be treated with such active substances, first of all, a “set of elements”, it is understood that the components (a) and (b) with activity included in the combination can be administered in the right doses independently of each other or different points in time using different fixed combinations that differ in the amounts of each of the components (a) and (b). Elements included in the "set of elements" can be entered simultaneously or sequentially, i.e. at different points in time and with the same or different time intervals for each element of the set of elements, preferably provided that the time intervals are chosen so that the effect on proliferative disease with the combined use of the elements is stronger than the effect that can be obtained by applying only one of the components (a) and (b) individually or when using both of them so that the compounds act independently of each other (for example, with enough pain with longer time intervals to avoid the influence of each of the components on the other component), this means that there is a stronger inhibition of proliferation or preferably a stronger regression or even cure of proliferative disease compared with the case when only one is administered separately at the same dose from components (a) and (b) or they are introduced at sufficiently large time intervals, such that the mutual effects of components (a) and (b) are excluded. This is indicated by the concept of "mutually enhance antiproliferative activity against proliferating cells, especially in the patient's body"; preferably this concept means a mutual enhancement of the action of components (a) and (b), especially synergism and / or causing regression of proliferating cells until their complete destruction and, above all, a pronounced synergistic interaction of components (a) and (b).

The term "proliferating cells" refers primarily to pathologically or abnormally proliferating cells, such as tumor cells and / or tumor metastases, especially tumors, listed above as preferred.

Preferred are combinations that have increased antiproliferative activity compared with the activity of each of the components individually, especially combinations that have a synergistic effect (synergistic combinations) or combinations that cause regression of proliferative tissues and / or cure for proliferative disease.

99-141 (1984)] и/или методом расчета коэффициента синергизма комбинации (КС) [Chou и др., Trends Pharmacol. Sci.

450-454 (1983); или Chou и др. New Avenues in Developmental Cancer Chemotherapy; Bristol-Myers Symposium Series, K.R. Harrap и I.A. Connors (ред.), 37-64, New York, Academic Press (1987)].The concept of "synergism" means an action in excess of the additive, i.e. a stronger effect of the combination of any component (a) with any component (b) than that which can be expected if the proliferation reduction coefficient is obtained only by multiplying the proliferation reduction coefficient for any component (a) taken separately or any component (b) taken separately, compared with the control without treatment, if each component (a) and (b), regardless of whether they are used individually or in combination, is administered in the same dose as in the case of a single treatment with one component ( it does not mean em, that the dose of component (a) should be identical to the dose of component (b), although this may also occur). Consider the following theoretical example, given only for the purpose of illustration: if component (a), when applied individually, causes a two-fold decrease in tumor growth (coefficient 2) compared to growth in the control without any treatment, and component (b) when its application individually causes a decrease in tumor growth corresponding to a coefficient of 1.5, then the additive effect with the combined use of component (a) and component (b) would lead to a 3-fold decrease in growth (multiplying 2 by 1.5). In the event that a more than 3-fold decrease in proliferation was detected, then this indicates the presence of a synergistic effect. The presence of synergism can be established by this method of fractional works [Webb in: "Enzymes and Methabolic Inhibitors", Volume 1, pp. 66-73 and 488-512, Academic Press, New York] or alternatively by the isobologram method [see links to: Berenbaum Oharmacol. Rev.

99-141 (1984)] and / or by the method of calculating the coefficient of synergism of combination (COP) [Chou et al., Trends Pharmacol. Sci.

450-454 (1983); or Chou et al. New Avenues in Developmental Cancer Chemotherapy; Bristol-Myers Symposium Series, KR Harrap and IA Connors (eds.), 37-64, New York, Academic Press (1987)].

The term "pharmaceutically acceptable carriers" is explained below in the definition of pharmaceutical compositions.

In the event that salt-forming groups are present in the corresponding molecule, component (b) (other chemotherapeutic agent (s)) may also be present in the form of salt (s), irrespective of whether it is specifically indicated above or below in the present description.

The treatment is preferably stopped in the event of one of the following situations: the disease progressively develops, for example, according to the reaction criterion of the Southwest Oncology Group (SWOG); an unacceptable level of toxicity has been detected (for example, for the patient, researcher, or both); 2 treatment cycles were carried out after the complete reaction was detected, for example, according to the reaction criterion of the Southwest Oncology Group (SWOG); or a patient rejection was received.

The salts of the components are primarily acid addition salts, base addition salts or, if several salt-forming groups are present, optionally also mixed salts or internal salts. Salts are primarily pharmaceutically acceptable salts, for example substantially non-toxic salts.

Such salts can form, for example, chemotherapeutic agents having an acidic group, for example a carboxy group, a phosphodiester or phosphorothioate group, and these salts are, for example, addition salts of suitable bases, such as non-toxic metal salts of groups Ia, Ib, IIa and IIb of the Periodic element systems, especially suitable alkali metal salts, for example lithium, sodium or potassium salts, or ammonium salts, as well as salts formed with organic amines, such as unsubstituted or substituted hydroxyl mono-, di- or trialkylamines, especially mono-, di- or tri (lower) alkylamines, or quaternary ammonium salts, for example, salts formed with N-methyl-N-ethylamine, diethylamine, triethylamine, mono- , bis- or tris (2-hydroxy (ness.) alkyl) amines, such as mono-, bis- or tris (2-hydroxyethyl) amine, 2-hydroxy-tert-butylamine or tris (hydroxymethyl) methylamine, N, N-di- (ness.) Alkyl-N- (hydroxy (ness.) Alkyl) amines, such as N, N-dimethyl-N- (2-hydroxyethyl) amine or three (2-hydroxyethyl) amine, or N -methyl-D-glucamine, or quaternary ammonium salts such as tetrabutylammonium salts. Chemotherapeutic agents having a basic group, for example an amino or imino group, can form acid addition salts, for example with inorganic acids, for example with hydrohalic acid, such as hydrochloric acid, with sulfuric acid or phosphoric acid, or with organic carbocyclic, sulfonic, sulfo or phosphoacids, or with N-substituted sulfamic acids, such as, for example, acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleino acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, gluconic acid, citric acid or benzoic acid, as well as amino acids, such as α-amino acids, as well as methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane -1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene-2-sulfonic acid, N-cyclohexyl sulfamic acid (with the formation of cyclamates) or with other acidic organic compounds eniyami such as ascorbic acid. Compounds having acidic and basic groups can also form internal salts. If more than one salt-forming group is present, the formation of mixed salts is also possible.

If concepts including numerical values are used above and below in the present description, it should be understood that they include numbers representing the upper and lower limits. For example, the term “from 1 to 3” means the range “from 1 inclusive to 3 inclusive” and the term “in the range from 1 to 3” means the range “from 1 inclusive to 3 inclusive”. All of the above applies to the case when instead of numbers (for example, 3), numerals (for example, "three") are used.

If the term “comprising” is used, then it can preferably be replaced by the term “consisting essentially of” or more preferably “consisting of”.

If the term “approximately” is used with reference to numbers, it preferably denotes a corresponding number of ± 15%, more preferably a number + 5%, most preferably simply a number without “approximately”. For example, "approximately 100" means "from 85 inclusive to 115 inclusive." If the term "approximately" is used in relation to a range of numbers, for example, "from about 1 to about 3" or "in the range from about one to about three," then preferably this means that the concept of "approximately" as defined in the previous sentence in relation to numbers , apply individually to each number denoting the beginning and end of the range. Preferably, if the term “approximately” is used with reference to any numerical values, then “approximately” may be omitted.

The term “weekly” means “about once a week” (this means that more than one treatment is performed with an interval between treatments of about one week), in which case “approximately” preferably means ± 1 day (i.e. “weekly” corresponds to “every 6-8 days”); more preferably “weekly” means “once every 7 days”.

The term “every 3 weeks” or “every three weeks” means “approximately once every three weeks” (this means that more than one treatment is carried out with an interval between treatments of approximately three weeks), in which case “approximately” preferably means ± 3 days (i.e., "every 3 weeks" corresponds to "every 18-24 days"); more preferably, “every 3 weeks” means “once every 21 days” (which is equivalent to the term “every third week”).

Preferred Embodiments

In the preferred embodiments described below, the general definitions may, if necessary, be replaced by more specific definitions given above and below in the present description.

(1) The present invention relates primarily to the treatment of a proliferative disease, in particular cancer, which cannot be treated with other chemotherapeutic agents and / or anti-cancer agents of the taxane class, especially with TAXOL®, more preferably one of the diseases mentioned above or below in the present description as preferred, according to which epothilone, preferably epothilone A or most preferably epothilone B, is administered to a person more than once, with an interval of one week to three weeks in a dose, which is calculated by the formula (I)

where N (integer or fractional number) denotes the number of weeks between treatments (from about one to about three weeks), i.e. N is from about 1 to about 3; more preferably, the dose for treatment is calculated according to formula II

even more preferably, the dose for treatment is calculated according to formula III

and most preferably, the dose for treatment is calculated according to formula IV

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

the introduction of epothilone, especially epothilone B is preferably carried out

(a) weekly to a person in a dose of from about 0.1 to about 6 mg / m ² , preferably from about 0.1 to about 5 mg / m ² , more preferably from about 0.1 to about 3 mg / m ² , still more preferably from 0.1 to 1.7 mg / m ² and most preferably from about 0.3 to about 1 mg / m ² ; or

(b) every three weeks to a person in a dose of from about 0.3 to about 18 mg / m ² , preferably from about 0.3 to about 15 mg / m ² , more preferably from about 0.3 to about 12 mg / m ² even more preferably from about 0.3 to about 7.5 mg / m ² , even more preferably from about 0.3 to about 5 mg / m ² and most preferably from about 1.0 to about 3.0 mg / m ² ;

the introduction is preferably carried out intravenously (iv) for 2-180 minutes, preferably for 2-120 minutes, more preferably for a period of time from about 5 to about 30 minutes, most preferably for a period of time from about 10 to approximately 30 minutes, for example, for approximately 30 minutes

(2) The present invention preferably also relates to the treatment of a disease associated with a tumor that cannot be treated with another chemotherapeutic agent, especially selected from the group consisting of 5-fluoracil and preferably a microtubule stabilizing agent from the taxane class, most preferably TAXOL®, wherein the tumor is selected from the group comprising a tumor of the gastrointestinal tract, for example, a colorectal tumor; swelling of the kidney; the genitourinary system, such as the prostate gland; tumor of the pancreatic gland; and a brain tumor (and / or any metastases thereof), most preferably a gastrointestinal tumor, preferably colorectal cancer, more preferably a gastrointestinal cancer, especially colorectal cancer, which cannot be treated with anti-cancer agents from the taxane class, especially such as TAXOL®, or most preferably a tumor that cannot be treated with standard chemotherapeutic methods, such as treatment with a standard chemotherapeutic agent, especially 5-fluorace silt; or a tumor of the genitourinary system, especially prostate cancer, most preferably prostate cancer that cannot be treated with hormones; according to which a warm-blooded animal, first of all, is administered epothilone A and / or B, primarily epothilone B.

(3) The present invention preferably also relates to the treatment of a disease related to a tumor, in particular a lung tumor, in particular non-small cell lung carcinoma, in particular lung cancer, which cannot be treated with anti-cancer agents of the taxane class, especially TAXOL®; breast cancer, especially multi-drug resistant; or an epidermoid tumor, preferably an epidermoid tumor of the head and neck, especially the mouth, especially if the latter has multiple drug resistance and / or resistance to treatment with anti-cancer agents from the taxane class, especially such as TAXOL®; according to which a warm-blooded animal, first of all, is administered epothilone A and / or B, primarily epothilone B.

(4) The present invention also preferably relates to an in vivo treatment regimen for a disease associated with a tumor, in particular (I) a tumor of the gastrointestinal tract, most preferably a tumor of the colon and / or rectum (colorectal tumor); and / or (II) a genitourinary tumor, especially a prostate tumor (preferably a prostate tumor that cannot be treated with hormones); first of all, in cases where such a tumor cannot be treated with other chemotherapeutic agents, primarily from the taxane class, most preferably such as TAXOL®, according to which epothilone A and / or B, first of all, epothilone B is administered to a person once at a dose in a range of from about 20 to about 100% of the MTD; and if necessary, one or more (preferably two to seven) additional doses, each of which is preferably in the dose range indicated above for the first dose, is administered during additional treatment cycles, preferably each dose is administered after a certain period of time that allows the treated the patient will recover sufficiently after the administration of each previous dose, preferably leaving more than two weeks after the previous treatment, more preferably after 2-10 weeks, most preferably 3-6 weeks after the previous treatment, especially three weeks after this treatment.

More preferably, epothilone B is administered to a person according to embodiments (1) to (4) weekly at a dose of from about 0.1 to about 6 mg / m ² , preferably from about 0.1 to about 5 mg / m ² , more preferably from about 0.1 to about 3 mg / m ² , even more preferably from 0.1 to 1.7 mg / m ^2, and most preferably from about 0.3 to about 1 mg / m ² ; or epothilone B is administered once every three weeks (every three weeks) at a dose of from about 0.3 to about 18 mg / m ² , preferably from about 0.3 to about 15 mg / m ² , more preferably from about 0.3 up to about 12 mg / m ² , even more preferably from about 0.3 to about 7.5 mg / m ² , even more preferably from about 0.3 to about 5 mg / m ² and most preferably from about 1.0 to approximately 3.0 mg / m ² . These doses are administered to a person, preferably intravenously (iv) for 2-180 minutes, preferably for 2-120 minutes, more preferably for a period of time from about 5 to about 30 minutes, most preferably for a period of time from about 10 to about 30 minutes, for example, for about 30 minutes

More preferably, this treatment is repeated every about 1 to about 3 weeks until progressive development of the disease is detected, an unacceptable level of toxicity is detected, 1 or preferably 2 treatment cycles are carried out after a complete reaction has been reached, or the patient has failed any reason.

(5) The present invention also preferably relates to an in vivo treatment regimen for a disease associated with a tumor, in particular (I) a tumor of the gastrointestinal tract, most preferably a tumor of the colon and / or rectum (colorectal tumor); and / or (II) a tumor of the genitourinary system, especially a tumor of the prostate gland; first of all, in cases where such a tumor cannot be treated with other chemotherapeutic agents, especially taxane agents, most preferably TAXOL® (preferably a prostate tumor that cannot be treated with hormones); according to which epothilone A and / or B, first of all, epothilone B is administered to a warm-blooded animal weekly at a dose of less than 80%, more preferably less than 50% of the maximum tolerated dose (MTD).

Preferably, in the case of weekly treatment of a person with said epothilone (s), the dose is in the range of from about 1 to about 60%, preferably from about 10 to about 60%, for example from about 5 to about 35% of the MTD, for example, in the range from about 30 to about 35% of the MTD. Preferably, when epothilone B is used in the treatment every three weeks, the dose is in the range of from about 5 to about 60%, preferably from about 10 to about 60%, even more preferably from about 10 to about 45%, most preferably from about 30 to about 45% of MTD. In some cases, the dose of epothilone B may be from about 2 to about 18 mg / m ² .

Most preferably, treatment is stopped after 3-8, especially 3-5 weekly administrations, followed by a rest period of 2-5, for example, 2 weeks, before resuming further processing. Preferably and if necessary, in the case of weekly treatment with epothilone B, it is stopped after 3-8 injections, followed by a period of rest for 2-4, for example 2 weeks, before resuming treatment by weekly administration.

(6) The invention also preferably relates to the treatment of an in vivo tumor-related disease by co-administering (a) epothilone A and / or epothilone B, especially epothilone B, in combination with (b) another chemotherapeutic agent selected from the group including:

(A) alkylating agents, preferably crosslinked chemotherapeutic agents, preferably bisalkylating agents;

(B) anti-cancer antibiotics, preferably doxorubicin (ADRIAMYCIN®, RUBEX®);

(B) antimetabolites;

(D) plant alkaloids;

(D) hormonal agents and antagonists;

(E) biological response modifiers, preferably lymphokines or interferons;

(G) protein inhibitors such as tyrosine kinases and / or serine / threonine kinases;

(H) antisense oligonucleotides or oligonucleotide derivatives;

or

(Ii) a variety of agents or other agents with a different or unknown mechanism of action;

the joint processing schedule is chosen so that component (a) and component (b) are used simultaneously or sequentially for a sufficiently short period of time, such that the active substances of both component (a) and component (b) can mutually enhance antiproliferative activity, for example, in the patient’s body.

(7) The invention also relates to a product for the treatment of a tumor associated with a disease, comprising component (a) and component (b), as defined in paragraph (6), in which one or more pharmaceutically acceptable carriers are present or may be absent, as a combined composition intended for simultaneous or sequential administration to humans over a sufficiently short period of time, such that the active substances of both component (a) and component (b) can mutually enhance the activity against disease associated with a tumor, especially (I) tumors of the gastrointestinal tract, most especially a tumor of the colon and / or rectum (colorectal tumor); and / or (II) a tumor of the genitourinary system, especially a tumor of the prostate gland; first of all, when such a tumor cannot be treated with another chemotherapeutic agent, especially an agent from the taxane class, most preferably such as TAXOL®.

In the embodiments indicated in paragraphs (1) to (7), or the embodiments described below, the administration of epothilone, especially epothilone B, is preferably carried out by infusion, especially by intravenous infusion.

Listed below are some of the most preferred embodiments of the invention:

A1. The use of epothilone, especially epothilone A and / or epothilone B for the treatment of proliferative disease, which cannot be treated with other chemotherapeutic agents; or the use of epothilone, especially epothilone B, for the preparation of a pharmaceutical composition intended for the treatment of a proliferative disease that cannot be treated with other chemotherapeutic agents.

A2. The use according to claim A1, wherein the proliferative disease is a disease associated with a tumor that cannot be treated with a microtubule stabilizing agent from the taxane class, especially such as TAXOL®.

A3. The use according to any one of claims A1-A2, wherein the proliferative disease is a colorectal tumor and / or its metastases.

A4. The use according to any one of paragraphs A1-A2, where the proliferative disease is a tumor of the prostate gland and / or its metastases; First of all, a tumor of the prostate gland that cannot be treated with hormones.

B1. The use of epothilone for the preparation of a pharmaceutical composition suitable for administration of a single dose of epothilone in the range of from about 1 to about 100% of the maximum tolerated dose (MPD) established for a warm-blooded animal, said warm-blooded animal to treat a proliferative disease that cannot be treated with other chemotherapeutic agents.

B2. The use according to claim B1, wherein the epothilone is epothilone A and / or epothilone B, preferably epothilone B.

B3. The use according to any one of paragraphs B1-B2, where the dose is in the range from 25 to 100% of the maximum tolerated dose and the warm-blooded animal is a human.

B4. The use according to any one of paragraphs B1-B3, where the standard dose of epothilone B for an adult, when treated once every three weeks, is in the range from about 0.3 to about 18, preferably from about 0.3 to about 12, more preferably from from about 0.3 to about 7.5, most preferably from about 1 to about 3 mg / m ² ; or during weekly treatment is in the range of from about 0.1 to about 6, preferably from about 0.1 to about 5, more preferably from about 0.1 to about 3, most preferably from about 0.3 to about 1 mg / m ² .

B5. The use according to any one of paragraphs B1-B4, wherein the dose is selected so that after a period of time necessary for sufficient recovery of the treated individual, the next dose may be administered after the previous dose has been administered.

B6. The use according to any one of claims B1-B5, wherein the proliferative disease is a tumor.

B7. The use according to any one of paragraphs B1-B6, where the proliferative disease is a colorectal tumor and / or its metastases.

B8. The use according to any one of claims B1-B6, wherein the proliferative disease is a tumor of the prostate gland and / or its metastasis.

B9. The use according to any one of Claims B1-B8, wherein the tumor is a tumor that cannot be treated with a microtubule stabilizing agent from the taxane class, especially such as TAXOL®.

IN 1. The use of epothilone, preferably epothilone A and / or B, in particular epothilone B, for the preparation of a pharmaceutical composition suitable for administration of epothilone once a week, where the dose is 80% of the MTD or less, preferably 50% of the MTD or less.

G1. The use of epothilone, preferably epothilone A and / or B, especially epothilone B, for the preparation of a pharmaceutical composition suitable for co-administration of (a) epothilone, preferably epothilone A and / or B, especially epothilone B, in combination with (b) another antitumor a chemotherapeutic agent to a warm-blooded animal that suffers from a proliferative disease that cannot be treated with other chemotherapeutic agents, especially a colorectal tumor or a prostate tumor and / or its metas the pelvis.

D1. A pharmaceutical composition intended for the treatment of a proliferative disease in a person, preferably a disease associated with a tumor, especially among those mentioned above and below in the present description as preferred, wherein the composition comprises epothilone, especially epothilone B, at a dose in a range from 1 to 100%, preferably from 25 to 100% of the maximum tolerated dose (MTD), and a pharmaceutically acceptable carrier.

E1. A combined composition comprising (a) epothilone A or preferably epothilone B and (b) one or more other antitumor agents and a pharmaceutically acceptable carrier.

G1. A product comprising, as component (a), epothilone A and / or B, preferably epothilone B, and as component (b), another antitumor agent, containing or not containing one or more pharmaceutically acceptable carriers, in the form of a combined composition intended for simultaneous or sequential administration to a warm-blooded animal, preferably a human, for a sufficiently short period of time, such that the active substances of both component (a) and component (b) can mutually reinforce the tumor activity in the body of a warm-blooded animal, intended for the treatment of proliferative disease.

Most preferably, the invention relates to the treatment of the following types of tumors / cancer using epothilone B:

(I) a tumor of the gastrointestinal tract, especially a colorectal tumor, which is not treatable by representatives of anti-cancer agents from the taxane class, in particular, such as TAXOL®, or more preferably, treatment using standard chemotherapeutic agents, especially such as 5- fluoracil and / or TAXOL®;

(Ii) a tumor of the genitourinary system, especially a tumor of the prostate gland, including primary and especially metastatic tumors; more preferably those that do not respond to hormone treatment;

(III) an epidermoid tumor, more preferably an epidermoid tumor of the head and neck, most preferably an epidermoid tumor of the mouth, especially a tumor that cannot be treated with other chemotherapeutic agents, primarily due to multiple drug resistance, especially treatment with representatives of taxan class anticancer agents, in particular, such as TAXOL®;

(IV) a lung tumor, especially non-small cell lung cancer, which cannot be treated with other chemotherapeutic agents, primarily (mainly) due to multiple drug resistance, especially treatment with representatives of taxanic class anti-cancer agents, in particular TAXOL®; and / or

(V) a breast tumor, especially a breast tumor that is resistant to many drugs, more preferably a tumor that is not treatable by representatives of anti-cancer agents from the taxane class, in particular such as TAXOL®.

Preferably, the invention relates to the treatment of any of the types of tumors listed in (I) to (V) above, most preferably the tumors indicated in (I), (II), (IV) and (V).

More preferably, the invention relates to the treatment of any types of tumors listed above in (I) to (V), preferably any of them, by intravenous infusion of epothilone B for 2-120 minutes, preferably for a period of time from about 5 to about 30 minutes more preferably for a period of time from about 10 to about 30 minutes, most preferably for about 30 minutes;

moreover, the introduction is repeated every one to three weeks, preferably every one week (weekly) or every three weeks;

wherein the dose of epothilone B is preferably calculated by the formula I

where N (integer or fractional number) denotes the number of weeks between treatments, comprising from about 1 to about 3 weeks, this means that N is in the range from about 1 to about 3; and y is 6, preferably 5, more preferably 4.

More preferably, the dose for treatment is calculated according to formula II

still preferably according to formula III

and most preferably according to formula IV

where in each of formulas II-IV, N is from about 1 to about 3 (which corresponds to intervals from about 1 to about 3 weeks between treatments).

More preferably, in the case of weekly treatment, the dose is from about 0.1 to about 6 mg / m ² , preferably from about 0.1 to about 5 mg / m ² , more preferably from about 0.1 to about 3 mg / m, still more preferably from 0.1 to 1.7 mg / m ² and most preferably from about 0.3 to about 1 mg / m ² ; or in the case of treatment once every three weeks is from about 0.3 to about 18 mg / m ² , preferably from about 0.3 to about 15 mg / m ² , more preferably from about 0.3 to about 12 mg / m ² , even more preferably from about 0.3 to about 7.5 mg / m ² , even more preferably from about 0.3 to about 5 mg / m ² and most preferably from about 1.0 to about 3.0 mg / m ² .

More preferably, rest periods of more than one week may be required, more preferably from two to ten weeks, even more preferably from three to six weeks (especially in the case of weekly treatments), after the previous treatment, depending on the condition of the patient, for example, for 3, 4, 6, 8 or more cycles, to recover sufficiently after the previous treatment.

Most preferred are also processing conditions and compositions similar to those described in the examples.

Pharmaceutical Compositions

The present invention also relates to the use of epothilone A and / or B, in particular epothilone A or preferably epothilone B, for the preparation of a pharmaceutical composition for the treatment of a tumor-related disease, as defined above in the present description; or a pharmaceutical composition for treating a tumor-related disease comprising epothilone A and / or B, especially epothilone A, or preferably epothilone B, and a pharmaceutically acceptable carrier.

The invention also relates to pharmaceutical compositions comprising epothilone A and / or epothilone B, preferably epothilone B, intended for the treatment of a proliferative disease, especially related to a tumor of the disease, as defined above in the present description, and to the preparation of pharmaceutical compositions intended for such treatment .

Epothilone A and / or B can be used, for example, for the preparation of pharmaceutical compositions that include an effective amount of the active substance in combination or in admixture with a sufficient amount of inorganic or organic solid or liquid pharmaceutically acceptable carriers.

The invention also relates to a pharmaceutical composition suitable for administration to a warm-blooded animal, in particular humans, for the treatment of a proliferative disease, as defined hereinabove, comprising such an amount of epothilone A and / or B, especially epothilone B, that is effective for treating this proliferative disease, together with at least one pharmaceutically acceptable carrier.

The pharmaceutical compositions of the invention are compositions for enteral, e.g., nasal, rectal or oral, or preferably parenteral, e.g., intramuscular or intravenous administration to a warm-blooded animal (human or animal), comprising an effective dose of a pharmacologically active substance individually or together with a sufficient amount of a pharmaceutically acceptable carrier. The dose of the active substance depends on the type of warm-blooded animal, body weight, age and individual condition, individual pharmacokinetic parameters, the disease to be treated, and the route of administration; preferably, the dose is one of the preferred doses, as defined above, and it should be adjusted accordingly if treatment of children is contemplated.

Pharmaceutical compositions include from about 0.00002 to about 95%, primarily (for example, in the case of ready-to-use infusion solutions) from 0.0001 to 0.02% or (for example, in the case of concentrates for infusion) from about 0, 1% to about 95%, preferably from about 20% to about 90% of the active substance (in each case, in wt.%). The pharmaceutical compositions of the invention may be in unit dosage forms, such as ampoules, vials, suppositories, dragees, tablets or capsules.

Preferably, the dose is selected so that it is possible to implement treatment regimens based on the above MTD for single or multiple administration in order to treat a disease associated with a tumor.

The pharmaceutical compositions of the present invention are prepared in a well-known manner, for example using conventional methods of dissolution, lyophilization, mixing, granulation or confection (assembly).

Solutions of the active substance are preferably used, as well as suspensions, and in particular isotonic aqueous solutions or suspensions, for example, in the case of lyophilized compositions which include the active substance individually or in combination with a pharmaceutically acceptable carrier, for example mannitol, to prepare such solutions or suspensions before use. The pharmaceutical compositions may be sterile and / or may include excipients, for example preservatives, stabilizers, wetting agents and / or emulsifiers, solubilizers, salts for regulating the osmotic pressure and / or buffers, and they can be prepared in a well-known manner, for example, using conventional methods of dissolution or lyophilization. Such solutions or suspensions may include viscosity enhancers such as sodium carboxymethyl cellulose, carboxymethyl cellulose, dextran, polyvinylpyrrolidone or gelatin.

Suspensions in oil include, as an oil component, vegetable, synthetic, or semi-synthetic oils commonly used for injection purposes. First of all, liquid fatty acid esters comprising, as an acid component, a long chain fatty acid having from 8 to 22, preferably from 12 to 22 carbon atoms, for example lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, arachidonic acid, behenic acid, or the corresponding unsaturated acids, for example, oleic acid, elaidic acid, erucic acid, brassidinic acid or linol acid-hand if necessary additives as antioxidants, for example vitamin E, β-carotene or 3,5-di-tert-butyl-4-hydroxytoluene. The alcohol component of these fatty acid esters preferably has a maximum of 6 carbon atoms and is a mono- or polyhydroxy alcohol, for example a mono-, di- or trihydroxy alcohol, for example methanol, ethanol, propanol, butanol or pentanol, or their isomers, but preferably glycol and glycerol. In particular, the following fatty acid esters can be noted as examples: ethyl oleate, isopropyl myristate, isopropyl palmitate, Labrafil M2375 (polyoxyethylene glycerol trioleate, Gattefosse, Paris), Miglyol 812 (triglyceride of saturated fatty acids having a chain length of 8-12 carbon atoms, Huls AG, Germany), and especially vegetable oils such as cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil and especially peanut oil.

Compositions for injection or infusion are prepared according to conventional methods under sterile conditions; the same applies to filling ampoules or vials with compositions and to sealing containers.

Preferred is an infusion composition that contains epothilone A and / or epothilone B, especially epothilone B, and a pharmaceutically acceptable organic solvent.

The composition does not require the use of a surfactant. Surfactants such as Cremophor can cause allergic reactions and they can also leach plasticizers from standard containers, tubes, etc. made of PVC. Therefore, when using them, it is necessary to use special devices for infusion, for example, nitroglycerin tubes and plastic-free containers, such as glass tubes, etc.

The pharmaceutically acceptable organic solvent used in the composition of the invention may be selected from any such solvents that are known to those skilled in the art. The solvent is preferably selected from alcohol, for example absolute ethanol, or ethanol / water mixtures, more preferably from 70% ethanol, polyethylene glycol 300, polyethylene glycol 400, polypropylene glycol or N-methylpyrrolidone, preferably from polypropylene glycol or 70% ethanol, or most preferably it represents polyethylene glycol 300.

Epothilones can preferably be present in the composition in a concentration of from about 0.1 to about 100 mg / ml, more preferably from about 1 to about 100 mg / ml, even more preferably from about 1 to about 10 mg / ml (especially in concentrates for infusion).

Epothilone A and epothilone B can be used as pure compounds or as a mixture of epothilone A and B. Due to its higher antitumor activity, epothilone B can be used in the composition at a lower concentration than epothilone A. When used in pure form, it is preferable to use a concentration of epothilone A component of 5 to 100 mg / ml, preferably 10 to 50 mg / ml, while when used in pure form, epothilone B is preferably used in a concentration of 0.1 to 10, more preferably 1 to 10, still more preferably from 1 to 2 mg / ml (these numbers relate primarily to the infusion concentrate, which must be diluted appropriately before treatment, see below).

Compositions of this type are conveniently stored in vials or ampoules. As a rule, vials or ampoules are made of glass, for example, borosilicate or sodium-calcium glass. The vials or ampoules can have any volume that is commonly used in the art, preferably they are large enough to include 0.5-5 ml of the composition. The compositions retain storage stability for a period of time up to 12-24 months at a temperature of at least 2-8 ° C.

Before administration of epothilone to a patient, the compositions should be diluted with an aqueous medium suitable for intravenous administration.

Preferably, the infusion solution should have the same or substantially the same osmotic pressure as the total body water. Therefore, the aqueous medium preferably contains an agent for maintaining isotonicity, which gives the solution for infusion the same or practically the same osmotic pressure as the total body water.

The agent for maintaining isotonicity may be selected from any such agent known in the art, for example, mannitol, dextrose, glucose or sodium chloride. Preferably, the agent for maintaining isotonicity is glucose or sodium chloride. Agents to maintain isotonicity can be used in amounts that give the solution for infusion the same or almost the same osmotic pressure as the total body water. The exact amounts needed can be determined using a routine experiment and they depend on the composition of the solution for infusion and on the type of agent to maintain isotonicity. When choosing a specific agent to maintain isotonicity, it is necessary to take into account the characteristics of the active substance.

The concentration of the agent to maintain isotonicity in the aqueous medium depends on the type of particular agent used. When glucose is used, its concentration is preferably in the range from 1 to 5 wt.% / Vol., More preferably 5 wt.% / Vol. When the agent for maintaining isotonicity is sodium chloride, it is preferably used in amounts of up to 1 wt.% / Vol, preferably about 0.9 wt.% / Vol.

The infusion composition may be diluted with an aqueous medium. The amount of aqueous medium used as a diluent is selected depending on the desired concentration of epothilone in the infusion solution. The infusion solution is preferably prepared by mixing the above infusion concentrate in a vial or ampoule with an aqueous medium, adjusting the volume to 20-200 ml, preferably 50-100 ml, using an aqueous medium.

Solutions for infusion may contain other excipients, which are usually used in compositions for intravenous administration. Excipients include antioxidants.

Antioxidants can be used to protect the active substance from oxidative decomposition. Antioxidants may be selected from antioxidants known to those skilled in the art and suitable for intravenous formulations. The amount of antioxidant can be determined using a routine experiment. As an alternative to or in addition to the use of antioxidants, an antioxidant effect can be obtained by preventing contact of oxygen (air) with the infusion solution. Typically, this can be done simply by purging the container containing the infusion solution with an inert gas such as nitrogen.

Solutions for infusion can be obtained by mixing the composition, enclosed in an ampoule or vial, with an aqueous medium, for example with 5 wt.% / Vol. a solution of glucose in water for injection (VDI) in an appropriate container, for example, in a cylinder or vessel for infusion.

The prepared solution for infusion is preferably used immediately or within a short period of time after preparation, for example, for 6 hours

Containers for storing infusion solutions can be selected from any conventional containers whose materials do not react with the infusion solution. In particular, glass containers made from the above types of glass may be used, although it may be preferable to use plastic containers, for example plastic infusion bottles.

Plastic containers can primarily be containers of thermoplastic polymers. Plastics may additionally include additives such as, for example, plasticizers, fillers, antioxidants, antistatic agents and other additives known in the art.

Plastics suitable according to the present invention must be resistant to the elevated temperatures necessary for heat sterilization. Preferred plastic infusion bottles are those made from plastics such as PVC known to those skilled in the art.

Containers having a wide range of sizes may be used. When choosing the size of the container, first of all, one should take into account the solubility of epothilone in a water medium, the simplicity of its preparation and, if necessary, the container's storage capacity.

It is preferable to use containers that can hold from about 250 to 1000 ml of infusion solution, more preferably from about 50 to about 120 ml.

The infusion solutions act in the same way as the infusion solutions of the paclitaxel agent interacting with the microtubes, and they have valuable properties in the treatment of conditions for which paclitaxel can be used. For certain types of tumors, epothilones have stronger beneficial effects compared to paclitaxel.

Dosage forms are usually administered intravenously at a dose of up to 100 mg / m ² for epothilone A and up to about 18 mg / m ² for epothilone B. The exact doses needed and the duration of administration depend on the severity of the condition and speed of administration, preferably they correspond to the above . In that case, if the dose can be administered intravenously, then the dose and concentration in the blood can be precisely determined using known methods in vivo and in vitro.

Pharmaceutical compositions for oral administration can be prepared by combining the active substance with solid carriers, if desired, by granulating the resulting mixture and then treating the mixture, if desired or if necessary after adding the appropriate excipients, to obtain tablets, dragee cores or capsules. It is also possible to include them in plastic carriers that enable the active substance to diffuse or release in predetermined quantities.

Suitable pharmaceutically acceptable carriers are primarily excipients, such as sugars, for example lactose, sucrose, mannitol or sorbitol, cellulose-based preparations and / or calcium phosphates, for example tricalcium phosphate or calcium bicarbonate, and binders, such as starch pastes, for example, corn, wheat, rice or potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and / or polyvinyl pyrrolidone; and / or, if necessary, disintegrants, such as the above starches, as well as carboxymethyl starch, crosslinked polyvinylpyrrolidones, agar, alginic acid or a salt thereof, such as sodium alginate. The excipients are primarily flow improvers and lubricants, for example, such as silicic acid, talc, stearic acid or its salts, such as magnesium or calcium stearate, and / or polyethylene glycol. If necessary, appropriate coatings, optionally resistant to the action of gastric juices, are applied to the dragee cores, inter alia, using concentrated sugar solutions, which may include gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, or using coating solutions in acceptable organic solvents, or for the purpose of preparing gastric juice resistant coatings using solutions of acceptable cellulose preparations such as ethyl cellulose phthalate or hydroxy phthalate propyl methylcellulose. Capsules are dry-filled capsules consisting of gelatin and soft-coated capsules made from gelatin and a plasticizer such as glycerol or sorbitol. Capsules filled with dry matter may contain the active substance in the form of granules, for example, with fillers, such as lactose, binders, such as starches, and / or with lubricants, such as talc or magnesium stearate, and, if necessary, with stabilizers. In soft-coated capsules, the active ingredient is preferably dissolved or suspended in suitable oily excipients such as fatty oils, paraffin oil or liquid polyethylene glycols, stabilizers and / or antibacterial agents may also be added. Dyestuffs or pigments can be added to tablets or dragee coatings or capsule coatings, for example, for the purpose of identification or labeling of different doses of the active substance.

In the case of a combination with another chemotherapeutic agent, a specific composition is prepared, as described above, comprising two or more of the above components (a) and (b), or two or more separate compositions (for example, a set of elements), or used commercially available in the form of standard compositions known to a person skilled in the art, other chemotherapeutic agent (s).

Examples:

The following examples are given to illustrate the present invention and are not intended to limit its scope. First of all, it should be understood that the cell lines indicated in the present description do not limit the scope of the invention, since they are only representative representatives and can be replaced by other cell lines and tumor cells of which they are representative.

Preparation of Compound Solutions

A mother solution of epothilone B in DMSO with a concentration of 10 mg / ml was prepared and stored at -20 ° C. Aliquots were diluted with aqueous solutions to give a final concentration of DMSO 5 vol.%, Tween 80 (polyoxyethylene sorbitan monooleate; ICI Americas, Inc.) 0.05 vol.% And 95 vol.% Saline (0.9 vol.% NaCl) .

Cells and cell culture conditions

1309-1312 [1991]; J. Biol. Chem.

18031-18040 [1089]; Int. J. Cancer,

696-703 [1991]) и обусловленными глютатионом механизмами устойчивости (Int. J. Cancer, 49: 688-695 [1991]).The human colorectal adenocarcinoma cell line HCT-15 (ATCC CCL 225) obtained from the American Type Culture Collection (American Type Culture Collection, Rockville, MD, USA) was used, and the cells were cultured in vitro according to the supplier's recommendations. HCT-15 is a cell line similar to epithelial (Cancer Res. 39: 1020-1025 [1979]), which has multiple drug resistance due to overexpression of P-glycoprotein (P-gp, gpl70, MDR-1; Anticancer Res.

1309-1312 [1991]; J. Biol. Chem.

18031-18040 [1089]; Int. J. Cancer,

696-703 [1991]) and glutathione-induced resistance mechanisms (Int. J. Cancer, 49: 688-695 [1991]).

1345-1355 [1978]), которую выделяли из асцитической жидкости пациента, также характеризуется морфологией, сходной с морфологией эпителиальных клеток, и обычно рассматривается как чувствительная к лекарствам.The Colo 205 cell line is a human colon carcinoma cell line (ATCC No.CCL 222; see also Cancer Res.

1345-1355 [1978]), which was isolated from the ascitic fluid of a patient, is also characterized by a morphology similar to the morphology of epithelial cells, and is usually regarded as drug sensitive.

To create subcutaneous and orthotopic models in mice, a human prostate cancer cell line independent of androgen was used. The PC-3M human prostate metastatic carcinoma cell line was obtained from Dr. I.J. Fidler (MD Anderson Cancer Center, Houston, TX, USA) and it was cultured in Hama F12K medium supplemented with 7 vol.% Fetal bovine serum (FBS). The PC-3M cell line was obtained by isolating from liver metastases caused in mice deprived of hairline after intracranial injection of PC-3M cell lines [ATCC CRL 1435; American Type Culture Collection, Rockville, MD, USA)] and they could grow in MEM Needle medium supplemented with 10% fetal bovine serum, sodium pyruvate, interchangeable amino acids, L-glutamine, a double-strength 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 negative for the androgen receptor, as is the supposedly derived PC-3M cell line. The PC-3 cell line was obtained from ATCC (ATCC CRL 1435) and it corresponded to stage IV prostate adenocarcinoma isolated from a 62-year-old Caucasian man; cells have low activity against acid phosphatase and testosterone 5-a-reductase. Cells are almost triploid with a modal chromosome number of 62. No normal Y chromosomes were detected by Q-band analysis.

The human lung adenocarcinoma cell line A549 (ATCC CCL 185; isolated as an explant culture from lung carcinoma tissue in a 58-year-old Caucasian man) has an epithelial morphology and can synthesize lecithin with a large percentage of desaturated fatty acids using the cytidine-diphosphocholine pathway; in all metaphases, a subtelocentric marker chromosome was found, including chromosome 6 and the long arm of chromosome 1. Human breast carcinoma cell line ZR-75-1 (ATCC CRL 1500; isolated from malignant ascites effusion in a 63-year-old Caucasian woman suffering from ductal carcinoma infiltrating ); originates from the epithelium of the mammary gland; cells have estrogen and other steroid hormone receptors and have hypertriploid chromosome numbers.

117-126 (1985) и у Fojo А. и др., Cancer Res.

3002-3007 (1985)) и ее культивировали согласно описанному в литературе методу (Меуег Т. и др., Int. J. Cancer

851-856 (1989)). Линию клеток КВ-8511, так же как и КВ-31 выводили из линии клеток KB (АТСС), эти клетки представляют собой клетки эпидермальной карциномы человека; клетки линии KB-31 могут быть выращены в виде монослоя с использованием модифицированной по методу Дульбекко среды Игла (D-MEM), содержащей 10%-ную фетальную телячью сыворотку (ФТС) (фирма М.А. Bioproducts), L-глутамин (фирма Flow), пенициллин (50 ед./мл) и стрептомицин (50 мкг/мл (фирма Flow)); затем они растут с периодом удвоения 22 ч и их относительная эффективность посева составляет приблизительно 60%. Линия клеток KB-8511 представляет собой линию клеток, выведенную из линии клеток КВ-31 с использованием циклов обработки колхицином; эти клетки характеризуются приблизительно в 40 раз большей относительной устойчивостью в отношении колхицина, чем клетки линии КВ-31; они могут расти в тех же условиях, что и клетки линии КВ-31.The KB-8511 human epidermal (oral) cell carcinoma cell line (P-gp overexpressing cell line derived from the KB-31 epidermoid (oral) carcinoma cell line) was obtained from Dr. RM Baker, Roswell Park Memorial Institute (Buffalo, NY , USA) (see description by Akiyama et al., Somat. Cell. Mol. Genetics

117-126 (1985) and Fojo A. et al., Cancer Res.

3002-3007 (1985)) and it was cultured according to the method described in the literature (T. Meueg et al., Int. J. Cancer

851-856 (1989)). The KV-8511 cell line, like KV-31, was removed from the KB cell line (ATCC), these cells are human epidermal carcinoma cells; KB-31 cells can be grown as a monolayer using Dulbecco's modified Eagle medium (D-MEM) containing 10% fetal calf serum (FCS) (M.A. Bioproducts), L-glutamine (firm Flow), penicillin (50 units / ml) and streptomycin (50 μg / ml (Flow company)); then they grow with a doubling period of 22 hours and their relative sowing efficiency is approximately 60%. The KB-8511 cell line is a cell line derived from the KB-31 cell line using colchicine treatment cycles; these cells are characterized by approximately 40 times greater relative resistance to colchicine than cells of the KB-31 line; they can grow under the same conditions as cells of the KV-31 line.

More detailed characteristics of cell lines can be found in the ATCC catalog and references cited there or in other references cited above.

In accordance with the Budapest Treaty, in the German Collection of Microorganisms and Cell Culture (DSMZ, Mascheroder Weg lb, D-38124, Braunschweig, Germany) on February 20, 1998 the following cell lines were deposited, mentioned above, under the corresponding registration numbers listed below: RS-3M : DSM ACC2338; A-549: DSM ACC2337; KV-8511: DSM ACC2342.

In addition, in accordance with the Budapest Treaty, in the German Collection of Microorganisms and Cell Culture (DSMZ, Mascheroder Weg 1b, D-38124, Braunschweig, Germany) on December 1, 1998, the following cell lines were deposited, mentioned above, under the corresponding registration numbers below: ZR-75-1: DSM ACC2376; HCT-15: ACC2377.

The following describes the general methods that were used in the experiments. If specific conditions are indicated, they are more powerful than the general descriptions given in the following paragraphs:

Assays of anti-proliferative activity:

Antiproliferative activity assays were performed according to methods described in the literature (Int. J. Cancer 43, 851-856 (1989)). Briefly, the method was as follows. Cells were plated at a density of 1.5 × 10 cells / well in 96-well microtiter plates and incubated overnight. On day 1, compounds were added as a serial dilution. Then the plates were incubated for another 2-5 days until at least once there was a doubling of the number of cells (depending on the cell line), after which the cells were fixed with 3.3 vol.% Glutaraldehyde, washed with water and stained with 0 , 05 vol.% Methylene blue. After washing, the dye was eluted with 3 vol.% HCl and the absorbance was measured at 665 nm using a SpectraMax 340 instrument (Bucherer, Basel, Switzerland). IC ₅₀ values were determined using a computerized system (SoftPro software package, Bucherer, Basel, Switzerland) using the formula (OD _test -OP _source ) / (OD _control -OP _source ) · 100. The value of the IC ₅₀ represents the concentration of the drug, causing 50% cell death in the hole compared to control cultures (100%) after the end of the incubation period.

In vivo assay of antitumor activity against subcutaneous transplanted tumors

Female or male BALB / c nu / nu mice (devoid of hair) were kept under sterile conditions (10-12 mice per type III cell) with free access to food and water. The weight of the mice was 20-25 g at the time of tumor implantation. Tumors were created by subcutaneous injection of cells (at least 2 · 10 ⁶ cells in 100 μl PBS or medium) to carrier mice (hosts) (4-8 mice for one cell line). The resulting tumors were serially transplanted using at least three consecutive transplantations before treatment. Tumor fragments (approximately 25 mg each) were subcutaneously implanted into the left flank of animals using a size 13 trocar needle, and the mice were anesthetized with foren (Abbott, Sweden).

Tumor growth and body weight were evaluated once or twice a week. All treatments were performed by the intravenous route (iv) and started when the average tumor volume reached approximately 100-250 mm ³ depending on the type of tumor. Tumor volumes were determined using the formula (L · D · π) / 6 (see Cancer Chemother. Pharmacol. 24: 148-154, [1989]). When treated with epothilone B, the dose and frequency of administration were varied. Agents selected as standards were administered according to previously established optimal drug regimens. In addition, for the purpose of representing changes in tumor volumes during the treatment period, antitumor activity was expressed as T / C% (average increase in tumor volumes in treated animals divided by the average increase in tumor volumes in control animals multiplied by 100). Tumor regression (%) was represented as the ratio of the smallest average tumor volume to the average tumor volume at the start of treatment according to the formula: regression (%) = (1-V _con / V _beg ) · 100 (V _con denotes the average final tumor volume, V _beg represents the average tumor volume at the start of treatment). All animals in which the tumor reached a size exceeding approximately 1.5-2.5 cm ³ were euthanized. Details of the experiment are given below.

In vivo analysis of antitumor activity against orthotopically injected cells

954-957 (1992)). Обработку начинали на 14-й день после инъекции клеток. Измерения, сделанные в этот момент времени, показывали, что средний размер опухоли составлял ~20 мг. Эпотилон В вводили либо один, либо два раза в неделю. Мышей умерщвляли через 42 дня после инокуляции опухоли, после чего тщательно выделяли предстательные железы, отсекали от соединительной ткани и взвешивали. Мезенгиальные лимфатические узлы исследовали в отношении присутствия метастазов, отсекали от соединительной ткани и взвешивали.PC-3M human prostate carcinoma cells (1 · 10 ⁶ cells per 20 μl of phosphate-buffered saline) were injected into each animal by injection into the left ventricle of the prostate gland (n = 6-9 individuals / group) according to the method described in the literature ( see Stephenson et al., J. Natl. Cancer Inst.

954-957 (1992)). Processing began on the 14th day after cell injection. Measurements made at this point in time showed that the average tumor size was ~ 20 mg. Epothilone B was administered either once or twice a week. Mice were sacrificed 42 days after tumor inoculation, after which the prostate glands were carefully isolated, cut off from the connective tissue and weighed. Mesengial lymph nodes were examined for the presence of metastases, cut off from connective tissue and weighed.

Tumor weight and body weight were evaluated once or twice a week. All treatments were carried out by the intravenous route (iv) and they were started 14 days after the injection. When treated with epothilone B, the dose and frequency of administration were varied. Antitumor activity was expressed as T / C% (average tumor weight in treated animals divided by the average tumor weight in control animals multiplied by 100).

Statistical analysis

The main strategy of statistical analysis is to use tests using multiple comparisons to identify the statistical significance of differences between different treated groups and differences within the group (for example, compared with the start of treatment) to determine whether treatment causes a stable regression of the disease or tumor. Since the volumes of subcutaneous tumors do not obey the law of normal distribution, differences in the volumes of subcutaneous tumors between groups with different treatments were determined using unidirectional analysis of variance (ANOVA) of ordered data (rank sum criterion) using the non-parametric Kruskal-Wallis criterion and the statistical significance of differences between the processed groups and control groups were determined using the Dunnett test. Pairwise comparisons between all groups were carried out according to the Student-Newman-Keuls (SNK) method. The weights of the organs are not always distributed according to the normal law and they were analyzed either using nonparametric criteria (as described above) or they were converted to the normal form (log [organ weight]) and analyzed using the unidirectional analysis of variance and then using the Dunnett test (comparison with controls) or Tukey criterion (comparisons between groups). In the statistical analysis of the increments (Δ) of the tumor volumes, the unidirectional method of variance analysis of ordered data using the Kruskal-Wallis criterion was used, the comparison of the treated groups with the media treated with controls was performed using the Dunnett criterion. Differences in body weights between the treated groups and vehicle-treated controls were analyzed using paired t-criteria. To determine the significance of differences in mortality between groups, Fisher's ideal test was used. For all criteria, the significance level was taken equal to p <0.05, however, it should be noted that the required level of confidence of 0.8 was never achieved for such small sizes of samples. For the orthotopic model, Fisher's ideal test was used to determine whether the relative numbers of mice with metastases differ in the treated groups and in the controls. All statistical calculations were performed using the SigmaStat 2.0 software (Jandel Scientific).

Materials

Epothilone B isolated from Myxobacteria Soranium cellulosum cultures was obtained from the Biopharmaceuticals Production and Development Department of Novartis Pharma, Basel, Switzerland. TAXOL® (a paclitaxel composition for clinical use) was obtained from Bristol Myers Squibb (USA), paclitaxel from Calbiochem (USA), and 5-fluoracil (Fluo-uracil®) from Roche (Switzerland). Cell cultures were obtained from Integra BioSciences (Wallisellen, Switzerland). Liquid media, fetal bovine serum (PBS) and media additives were obtained from Gibco / BRL (Basel, Switzerland). BALB / c nu / nu mice (hairless) were obtained from Bomholtgaard (Copenhagen, Denmark) or obtained from Novartis vivarium (Sisseln, Switzerland). Conventional BALB / c mice were obtained from Iffa Credo (France) or from Novartis vivarium (Sisseln, Switzerland).

Determination of the maximum tolerated dose (MTD)

To determine the MTD of female BALB / c mice deprived of hair or BALB / c (obtained from vivarium from Novartis, Sisseln, Switzerland), epothilone B was treated once by intravenous injection (n = 3 for a single dose group). The dose was increased (2, 4, 6, 8, 10 and 12 mg / kg) and the mice were examined for the occurrence of pronounced toxic effects within 10 days after treatment with the drug.

Example 1: Determination of the maximum tolerated dose (MTD)

The results of experiments to determine the maximum tolerated dose (MTD) are presented in table 1 and 2.

Epothilone B was administered by a single iv. injections in the form of a bolus at a dose of 2, 4, 6, 8, 10 or 12 mg / kg. The daily survival and body weight of the mice were evaluated. Changes (Δ) in body weight were determined by comparing the last measured body weight with body weight before treatment.

Epothilone B was administered by a single iv. injections in the form of a bolus at a dose of 2, 4, 6, 8, 10 or 12 mg / kg. The daily survival and body weight of the mice were evaluated. Changes (Δ) in body weight were determined by comparing the last measured body weight with body weight before treatment.

The results of these experiments showed that for normal mice, the MTD was approximately 4 mg / kg, while for mice lacking hair, the MTD was approximately 6 mg / kg.

Example 2: Toxicity of epothilone B (comparative study of toxicity after intravenous administration to mice for two weeks)

In order to assess the subchronic toxicity of epothilone B upon intravenous administration, GLP toxicity was studied upon intravenous administration for two weeks to tumor-free normal female BALB / c mice, including analysis of mortality, clinical symptoms, body weight, amount of food ingested, hematological analysis, clinical biochemical analysis, analysis of urine and organ weight, as well as macro and microscopic studies. The study was carried out for two different dose regimes of epothilone B of 3 and 10 mg / kg, respectively, which were administered on day 1 and day 8 (8 animals in the group).

Half of the animals were killed on day 15 (main group) and an autopsy was performed. After administration of the second dose, the second half (recovery group) was kept for a recovery period of 5 weeks, after which it was sacrificed and autopsy was performed on day 43. However, all animals in the recovery group, treated with a dose of 10 mg / kg, had to be killed prematurely on day 19 due to the poor general condition of the body.

There were no deaths recorded during the entire treatment period for all dose levels (days 1-14), and in the recovery group treated with a dose of 3 mg / kg, all animals survived until the end of the experiment. During the first and second weeks, body weight loss was observed in all animals in the group treated with a dose of 10 mg / kg, while when treated with a dose of 3 mg / kg, body weight loss became noticeable only in the second week. The increase in body weight during the recovery period in the treated and control animals was similar.

Both dose levels of epothilone B induced a decrease in the number of leukocytes, primarily neutrophils and lymphocytes, in all treated animals (day 15), however, this effect was more pronounced for a dose of 10 mg / kg. In addition, when treating with both dose levels in individual animals, there was a slight increase in the number of basophils, as well as a decrease in monocyte levels. A slight decrease in erythrocyte parameters, accompanied by an increase in reticulocytes and platelets, was observed only for a dose of 10 mg / kg. After the recovery period (day 43), the hematological parameters returned to normal in two of the four animals in the group treated with a dose of 3 mg / kg, while the other two still had reduced white blood cell counts.

Only minor changes were observed in the clinical chemical profile of the treated animals, which could not be unambiguously correlated with treatment with epothilone B.

Treatment with epothilone B for both dose levels resulted in marked changes in the weight of the thymus, spleen and uterus (day 15). In addition, slight decreases in liver weight were observed. (The weight of the organ was determined for the adrenal glands, liver, thymus, spleen, brain, ovaries, kidneys, uterus, and heart). When treated with a dose of 3 mg / kg, the weight of the organs after the recovery period (day 43) turned out to be comparable with the weight of the organs of control animals, which indicated complete recovery. (In euthanized animals treated with a dose of 10 mg / kg, no organ weight was evaluated).

Microscopic examination after histological processing of the isolated tissues in animals euthanized on day 15 revealed the average or severe atrophy of the thymus when treated with doses of 3 and 10 mg / kg, respectively. In addition, when treated with a dose of 10 mg / kg, minimal lymphoid atrophy in the spleen was observed, from minimal to weak myeloid hypoplasia in the bone marrow of the sternum and minimally increased hematopoiesis in the spleen. When treated with a dose of 3 mg / kg, erythroid and myeloid atrophy was observed from minimal to weak sternum in the bone marrow. For both dose levels, minimal necrosis of single cells in the intestinal mucosa (colon and small intestine) was noted, but necrosis was more pronounced for a dose of 10 mg / kg.

In animals from the recovery groups, weak myeloid hyperplasia and / or bone marrow atrophy were observed for both dose levels. In the case of treatment with a dose of 10 mg / kg, a weak lymphoid atrophy was also found in the spleen, as well as mild to moderate hemosiderosis. For a dose of 10 mg / kg, it was not possible to isolate the thymus tissue for microscopic examination, which indicated that thymic atrophy could also occur in these mice. In animals from the recovery group treated with a dose of 3 mg / kg, no histological changes in the thymus were detected on day 43.

In conclusion, it should be noted that epothilone B at a dose of 3 mg / kg (administered on day 1 and day 8) was tolerated without death during the entire observation period of 43 days, while animals treated with a dose of 10 mg / kg were necessary kill on day 19 due to poor health. During the treatment period, weight loss occurred for both dose levels. Hematological analysis revealed lower levels of leukocytes, neutrophils and lymphocytes in both groups of individual mice, as well as increased levels of basophils and decreased levels of monocytes. In addition, signs of anemia were detected for both dose levels. No effect on the clinical chemical profile was found. Only after the end of the treatment period (day 15) with doses of 3 and 10 mg / kg, mild to moderate thymus atrophy was observed, and for a dose of 10 mg / kg also minimal lymphoid atrophy in the spleen was observed. In addition, for a dose of 10 mg / kg, it was possible to detect myeloid hypoplasia in the bone marrow and increased hematopoiesis in the spleen. For a dose of 3 mg / kg, weak erythroid and myeloid atrophy of the bone marrow was manifested. For doses of 3 and 10 mg / kg, necrosis of individual cells in the intestinal mucosa was detected only after the end of the treatment period.

conclusions

The most important conclusions based on the data summarized in examples 1 and 2 regarding the toxicological characteristics of epothilone B can be summarized as follows:

The magnitude of the MTD at r.v. the introduction of a single dose of epothilone B for normal and hairless mice of the BALB / c main line are 4 and 6 mg / kg, respectively. Thus, hairless mice were less sensitive to the toxic effects of the compound compared to normal mice.

For normal mice, two doses of 3 mg / kg, administered at intervals of one week, were fairly well tolerated and did not cause death until day 43 after the first dose. A dose level of 10 mg / kg under the same dosage regimen led to the death (or killing) of all treated animals.

Example 3: In vitro activity of epothilones against cell lines

For some human cancer cell lines, a pronounced antiproliferative activity of epothilone B was detected; the results of these experiments are summarized in table 3. In general, epothilone B has a higher activity than paclitaxel, especially for cancer cells with the multiple drug resistance (MDR) phenotype (for example, such as RD-8511, HCT-15).

Comparison of IC ₅₀ [nM] values when inhibiting the growth of human carcinoma cell lines using epothilone B and paclitaxel (exposure for 5 days, average values ± SD, n = 3).

The values in parentheses characterize relative stability, i.e., IC ₅₀ (stable line) / IC ₅₀ (parent line).

Example 4: Antitumor activity of epothilones against the tumor cell line HCT-15 of human colorectal adenocarcinoma

When they were used individually or in combination, tumor volumes were used as the main indicator of the activity of antitumor agents, and changes in body weight were determined as an indicator of treatment tolerance.

As can be seen from table 4, a single dose of 4 mg / kg of epothilone B can cause tumor regression (p <0.05 when compared with controls; Dunnett's test) in drug-resistant overexpressing P-gp colorectal tumor cells of the HCT-15 line ( figure 1 and table 1). This activity is clearly superior to the effect caused by five-fold administration of TAXOL® at a dose of 20 mg / kg or two-fold administration of 5-fluoracil at a dose of 75 mg / kg (p <0.05 when compared with epothilone B; SNK criterion). Tumors of the HCT-15 line are resistant to both TAXOL® and 5-fluoracil, which was expressed in the fact that for these drugs, final T / C values of 50 and 88% were obtained, respectively (in both cases, p> 0.05 at comparison with controls; Dunnett's test). Treatment with epothilone B proved to be well tolerated, this was expressed in the fact that the body weight remained constant during treatment; in mice treated with the vehicle, weight gain was observed. When treated with epothilone B, no deaths were observed. In contrast, when processing using TAXOL®, several deaths were recorded (1/8 [12.5%] deaths), while treatment with 5-fluoracil revealed a higher level of deaths (4/8 [50%] deaths); however, due to the small number of treatment groups, no statistical reliability of the results was achieved (p> 0.05 when compared with controls; Fisher's ideal test). Mice that survived treatment were characterized by stable body weight.

These results indicate that epothilone B has a pronounced antitumor effect against tumors of the HCT-15 line, resistant to TAXOL® and 5-fluoracil, and is also well tolerated in the studied dose of 4 mg / kg.

Tumor fragments weighing approximately 25 mg were implanted in the left flank of each of the female hairless mice (n = 8 in the group). Treatments began on day 14 after tumor transplantation. Epothilone B was administered on day 14 by a single iv. injections at a dose of 4 mg / kg. 5-Fluoracil was administered on day 14 and day 21 by iv. injection at a dose of 75 mg / kg. TAXOL® was administered by iv injections at a dose of 20 mg / kg / day every second day for 5 treatment cycles (days 14, 16, 18, 20 and 22). Antitumor activity was expressed as T / C% (average increase in tumor volumes in treated animals divided by the average increase in tumor volumes in control animals multiplied by 100). Tumor regression (%) was characterized by comparing the final average tumor volume with the average volume at the start of treatment. The increment (Δ) of the tumor volume is the tumor volume on the last day of treatment minus the tumor volume on the first day of treatment. Statistical analysis of increments Δ of tumor volumes was performed using the Dunnett test by comparing the treated groups with controls. Statistical analyzes of changes in body weight were performed using paired t-criteria by comparing body weights before treatment with the corresponding values obtained after treatment; the weight of the mice at the beginning of the treatment was ~ 20-25 g. The abbreviation "n.d." stands for "statistically unreliable." Data are mean ± SD obtained for animals that survived the end of the experiment.

Example 5: Comparison of the antitumor activity of epothilone B and TAXOL® against epidermoid carcinoma of the human KB-8511 line, subcutaneously transplanted to female BALB / c mice deprived of hairline

As can be seen from the data presented in table 5, different dose regimes of epothilone B allowed to inhibit the growth of TAXOL® resistant tumors of the KB-8511 line in mice without hairline. A single injection of 4 mg / kg epothilone B caused a short-term regression (-51% on day 25 after transplantation; p <0.05 when compared with vehicle-treated controls, Dunnett's test), however, the tumors began to grow again, resulting in day 40 after treatment, the final T / C value was 24% (p <0.05 when compared with controls treated with the vehicle, Dunnett's test). Such a single administration of epothilone B was well tolerated, while maintaining a stable body weight and no death of animals.

Weekly intravenous administration of epothilone B led to inhibition of tumor growth depending on the dose: a dose of 4 mg / kg caused 98% regression (p <0.05 when compared with vehicle-treated controls, Dunnett's test); a dose of 2 mg / kg caused a short-term 44% regression and gave a final T / C value of 14% (for both characteristics, p <0.05 when compared with vehicle-treated controls, Dunnett's test); a dose of 1 mg / kg gave a final T / C value of 81% (p <0.05 when compared with controls treated with a carrier, Dunnett's test). TAXOL® did not have activity against tumors of the KV-8511 line (T / C was 132%, p <0.05 when compared with vehicle-treated controls, Dunnett's test). After the experiment, 5 out of 8 mice treated with a dose of epothilone B 4 mg / kg / week and 1 out of 8 mice treated with a dose of 2 mg / kg / week, no tumors were detected. Although a weekly treatment with a dose of 4 mg / kg tended to decrease body weight (-5 ± 7%), it was not statistically significant. In all control groups treated with the carrier, and in all groups treated with epothilone B at doses of 2 and 1 mg / kg / week and treated with TAXOL®, an increase in body weight was observed, while there were no deaths, indicating good tolerability of the treatments.

These results indicate that epothilone B has activity against experimentally induced TAXOL® resistant epidermoid tumors.

Tumor fragments weighing approximately 25 mg were implanted in the left flank of each of the female hairless mice (n = 8 in the group). Treatments began on day 13 after tumor transplantation. Epothilone B was administered on day 13 by a single dose. in. injections at a dose of 4 mg / kg or once a week by iv injections in doses of 4, 2 or 1 mg / kg (on days 13, 21 and 27). TAXOL® was administered by iv injections at a dose of 20 mg / kg / day every second day for 5 treatment cycles (days 13, 15, 17, 19 and 21). Antitumor activity was expressed as T / C% (average increase in tumor volumes in treated animals divided by the average increase in tumor volumes in control animals multiplied by 100). Tumor regression (%) was characterized by comparing the final average tumor volume with the average volume at the start of treatment. Changes (Δ) in tumor volumes represent tumor volume on the last day of treatment minus tumor volume on the first day of treatment. Statistical analysis of changes in Δ tumor volumes was performed using the Dunnett test by comparing the treated groups with controls. Statistical analyzes of changes in body weight were performed using paired t-criteria by comparing body weights before treatment with the corresponding values obtained after treatment; the weight of the mice at the beginning of the treatment was ~ 20-25 g. The abbreviation "n.d." stands for "statistically unreliable." Data are mean ± SD obtained for animals that survived the end of the experiment.

The experimental results showed that while TAXOL® was ineffective, treatment with epothilone B showed the presence of pronounced antitumor activity; when using a dose of 4 mg / kg, regression could even be achieved.

Example 6: Antitumor activity of epothilones against orthotopically injected prostate tumor cells of the PC-3M line

The results characterizing the activity of epothilone B against tumors of the PC-3M line, which initially grows in the prostate gland, and then forms metastases in the mesengial lymph nodes, are presented in table 6.

In this experimental model of prostate cancer, PC-3M cells initially grew in the prostate and then metastasized to the mesengial lymph nodes. Organ weight measurements were used to evaluate the antitumor activity of the treatments.

Table 6 presents data on the development of experimentally induced cancer, starting from day 14 after the injection of the cells (the beginning of the treatment), when there was no disease of the lymph nodes, until day 42 after the injection of the cells, when the prostate and mesenteric lymph nodes sharply increased their weight . All modes of administration of epothilone B were highly effective (in all cases p <0.05 when compared with controls treated with a carrier, Dunnett's test for logarithmic data) for reducing prostate weight and preventing the development of tumor metastases in mesenteric lymph nodes. The antitumor effects of all treatments with active ingredients were found to be equivalent (p> 0.05: Dunn test). In each of the groups treated with epothilone B, metastases were found in only one animal, while they were found in all control animals treated with a vehicle (p <0.05; Fisher's ideal test), which indicates that treatment with epothilone B significantly reduced the formation of detectable metastases.

Epothilone B treatments were not very well tolerated in high doses. While in the case of a single dose of 6 mg / kg or two doses of 4 mg / kg, only a tendency to decrease in body weight could be detected, in the case of a single dose of 8 mg / kg or weekly 5 mg / kg significant reduction in body weight (table 6). Apparently, the treatments contributed to the survival of mice having a tumor; however, probably, due to the small numbers of animals in each of the treated groups, statistically reliable results were obtained only for the treatment regimen with a dose of 6 mg / kg (once) (p = 0.029, Fisher's ideal criterion for the final numbers of surviving animals).

The results of studies using this model indicate a high antitumor activity of epothilone B both in relation to the reduction of primary tumors and in preventing the development of metastases. However, for some treatment modes, epothilone B was poorly tolerated.

The results of the study showed that epothilone B has activity against human prostate carcinoma both in vitro and in vivo (see example 3). Epothilone B also has the ability to reduce the growth of the primary tumor and strongly inhibit the formation of detectable metastases in the orthotopic model of prostate cancer. In addition, it can also contribute to the survival of mice with a tumor, however, this requires additional experimental study. Along with the pronounced antitumor activity of epothilone B treatment, significant weight loss was caused for the studied drug regimens. However, the reasons for this poor tolerance are unknown.

Of particular note is the activity of epothilone B shown in the orthotopic tumor model of PC-3M. Orthotopic models are created by implanting a tumor in the tissue in which the primary tumor develops in humans, and unlike subcutaneous injection models, metastases often occur in this case. Therefore, the fact that epothilone B inhibits the growth of a primary tumor in the prostate gland and inhibits the formation and / or growth of metastases in the mesenteric lymph nodes indicates a significant activity of epothilone B.

In general, due to the pronounced antitumor activity demonstrated in experiments on models of prostate cancer that are considered relatively resistant to chemotherapy (Br.J. Cancer 75. 1593-1600 (1997)), epothilone B appears to be a promising agent for the treatment of prostate cancer.

1 · 10 ⁶ Cells of the PC-3M line in 20 μl PBS were injected into the left ventricle of the prostate gland of each of the male hairless mice (n = 9 in the group). Treatments began on day 14 after injection of tumor cells. Epothilone B was administered by iv injections once at a dose of 6 or 8 mg / kg, or once a week at a dose of 4 or 5 mg / kg. Antitumor activity was expressed as T / C% (average tumor weight in treated animals divided by the average tumor weight in control animals multiplied by 100). Changes in body weight were evaluated only in animals that survived to the end of the experiment (day 42). Statistical analyzes of changes (Δ) in body weight were performed using paired t-criteria by comparing body weights before treatment with the corresponding values obtained after treatment; the weight of the mice at the beginning of the treatment was ~ 20-25 g. The abbreviation "n.d." stands for "statistically unreliable."

Example 7: Activity of epothilone B against non-small cell lung carcinoma A549

3-10 Million cells were implanted subcutaneously in the right axillary (lateral) region of thymus-free mice obtained by outbreeding (nu / nu), and grown until the tumor volume reached approximately 100 mm ³ . Epothilone B was dissolved in 1% DMSO in a 5% glucose solution in distilled water (D5W) and iv was administered. either only once, or once a week, or 3 times a week, or 5 times a week. For positive controls, TAXOL® clinical formulations diluted 6 times with D5W, which were administered iv, were used. 3 times a week.

Antitumor activity was expressed as T / C% (comparing changes (Δ) in tumor volumes in the treated group and the control group treated with the vehicle) after the experiment. Regression was evaluated by the formula: - (T / T ₀ -1) · 100%, where T is the tumor volume in the treated group at the end of the experiment, at ₀ is the tumor volume at the beginning of the experiment. Statistical validity was evaluated using the one-way Student t-test.

Results: table 7 summarizes the results of studies for tumors line A549. When administered once at a dose of 6 mg / kg, epothilone B caused significant inhibition of the tumor (T / C = 41%), while toxicity was not detected. A dose of 4 mg / kg, administered once a week (4 mg / week), caused the tumor to stop growing (T / C = 7%), but at the same time led to a 13% loss in body weight. For comparison, when using a dose of 1.5 mg / kg administered 3 times a week (4.5 mg / week), all animals had to be euthanized in the first week of the experiment due to the detected toxicity. Doses of 0.5 mg / kg administered 5 times a week (2.5 mg / week) caused tumor inhibition identical to the inhibition for a single administration regimen (T / C = 41%), however, apparently due to accumulation toxicity occurred 23% weight loss. TAXOL®, administered 3 times a week at a dose of 20 mg / kg, did not lead to inhibition of tumor growth, but caused 16% loss of body weight, with a mortality rate of 1 case per 8 mice.

Treatments began on day 16 after implantation (10 million cells / individual). Epothilone was administered iv 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 a dose of 1.5 mg / kg (days 16, 18, 21 , 23, 25, 28, 30, 32 and 35) or five times a week at a dose of 0.5 mg / kg (days 16-18, 21-25, 28-32 and 35-36). TAXOL® was administered iv three times a week in an amount of 20 mg / kg, divided into doses of 10 mg / kg, which were administered with an interval of 1 hour (days 16, 18, 21, 23, 25, 28, 30, 32 and 35). The media as a control (1% DMSO / D5W) was administered iv. three times a week (days 16, 18, 21, 23, 25, 28, 30, 32 and 35). All final data were recorded on day 37. One asterisk denotes p <0.05, and a double asterisk denotes p <0.01, and p values were obtained using the one-way Student t-test. "but." means “not determined” (animals were euthanized because toxicity of the compound was detected).

Example 8: Comparison of the antitumor activity of epothilone B and TAXOL® against non-small cell tumors of the light line A549 in mice lacking hair

The materials and methods described above were used for the xenograft model of a human tumor. 10 Million cells (lines A549) were implanted subcutaneously in the right axillary (lateral) region of thymus-free mice obtained by outbreeding (nu / nu), and grown until the tumor volume reached approximately 100 mm ³ . Epothilone B was dissolved in 1% DMSO in a 5% glucose solution in distilled water (D5W) and iv was administered. once a week for 3 weeks. For positive controls, TAXOL® clinical formulations diluted 4 times with D5W, which were administered iv, were used. 3 times a week for 3 weeks in divided doses (2 × 10 mg / kg) with an interval of 1 hour.

Antitumor activity was expressed as T / C% (comparing changes (Δ) in tumor volumes in the treated group and the control group treated with the vehicle) at the end of the experiment. Regression was evaluated by the formula: - (T / T ₀ -1) · 100%, where T is the tumor volume in the treated group at the end of the experiment, and T ₀ is the tumor volume at the beginning of the experiment. Measurements were continued for another 2 weeks after completion of the regular 3-week experiments to evaluate the reversibility of drug-induced weight loss and preservation of antitumor effects. Statistical significance was evaluated using the one-way Student t-test, Dunnett or Dunn criteria.

Results: Table 8 summarizes the results of standard 3-week experiments obtained for tumors of the A549 line. The introduction of epothilone Once a week caused a statistically significant inhibition of tumor growth depending on the dose, tumor growth was stopped using the maximum concentration of the drug. When administered at doses of 3.5 and 3 mg / kg, epothilone caused a marked inhibition of tumor growth (T / C = 15 and 23%, respectively). Both doses caused a noticeable but reversible (see table 8) body weight loss of approximately 15%. Doses of 2 and 1 mg / kg gave T / C values of 43 and 74%, which were statistically significant, when using a dose of 2 mg / kg, there was no change in body weight, and when using a dose of 1 mg / kg, a normal increase was observed body weight. TAXOL® when administered 3 times a week in divided doses (2 · 10 mg / kg) did not cause inhibition of tumor growth, but led to a 19% loss in body weight.

Treatments began on day 13 after implantation (10 million cells / individual). Epothilone B was administered iv once a week for 3 weeks (days 13, 20 and 27) in doses of 1, 2, 3 and 3.5 mg / kg. TAXOL® was administered iv three times a week for 3 weeks (days 14, 17, 19, 21, 24, 26, 28, 31 and 33) in an amount of 20 mg / kg in divided doses of 10 mg / kg with an interval of 1 hour. Final data recorded on day 34. One asterisk indicates p <0.05 obtained using the one-tailed Student t-test, and a double asterisk indicates p <0.05 obtained using the Dunnett or Dunn test.

The measurements were carried out for another 2 weeks after completion of the regular 3-week experiments and the final data obtained after 5 weeks are summarized in table 9. The antitumor effect remained unchanged, while the animal body weight was restored. T / C values for epothilone B dose levels of 3.5, 3, 2, and 1 mg / kg were 12, 16, 49, and 72%, respectively, and normal weight gain was observed in all groups. TAXOL® remained ineffective and only 2% weight gain was observed in animals.

Measurements for the experimental conditions described in table 1 were continued for another two weeks. Final data were recorded on day 48. One asterisk indicates p <0.05 obtained using the one-tailed Student t-test, and a double asterisk indicates p <0.05 obtained using the Dunnett or Dunn test.

Example 9: Antitumor effect of epothilone B against breast tumors of the line ZR-75-1

Table 10 shows the results of an experiment that compared the effects of TAXOL® and epothilone B on the breast cancer cell line ZR-75-1. The methods used for this tumor model are described above.

The results allow us to conclude (based on antitumor efficacy) that the best dose regimen is a weekly administration of 4 mg / kg. However, for all doses, cases of animal death were observed, which indicated that the ZR-75-1 line tumor, unlike other tumor types, could affect the general health of mice.

Tumor fragments weighing approximately 25 mg were implanted in the left flank of each of the female hairless mice (n = 6 in the group); Pelletized estrogen was injected subcutaneously into the opposite side. Treatments began on day 19 after transplantation. Epothilone B was administered by iv injections at a dose of 1, 2 or 4 mg / kg, either once a week (days 19, 26 and 33), or every second week (days 19 and 33). The data presented are obtained for animals surviving by day 47, the last day of observation. Antitumor activity was expressed as T / C% (average increase in tumor volumes in treated animals divided by the average increase in tumor volumes in control animals multiplied by 100). Tumor regression (%) was evaluated by comparing the final average tumor volume with the average tumor volume at the beginning of the experiment. Changes (Δ) in tumor volume represent tumor volume on the last day of treatment minus tumor volume on the first day of treatment.

Example 10: Comparison of the antitumor effect of epothilone B and 5-fluoracil against subcutaneous transplanted colon tumors of Colo 205

Table 11 presents data on the effect of epothilone B against subcutaneously transplanted tumors of the Colo 205 line, as well as on the effect of 5-fluoracil. In contrast to the treatment of the tumor cell line HCT-15, where standard treatment with 5-fluoracil or treatment with TAXOL® was ineffective, in this case it turned out that treatment with 5-fluoracil was still effective, although less effective than treatment with epothilone B.

Along with the results shown in example 4, indicating that the PCT-15 cell line does not respond (cannot be influenced) to either TAXOL® or the standard treatment for colon cancer with 5-fluoracil, these data show that epothilone In fact, it can be used to treat tumors that are not amenable to known standard methods of treatment. On the other hand, it is also more effective in cases where standard treatments are also effective. As a preferred dosage regimen, a schedule may be proposed involving 4 mg / kg every 2 weeks (tumor regression, no animal deaths). Such treatment is even more preferable compared to treatment with 5-fluoracil, in which no regression was detected, but only 4 out of 7 animals survived.

Tumor fragments weighing approximately 25 mg were implanted in the left flank of each of the female hairless mice (n = 7 in the group); Pelletized estrogen was injected subcutaneously into the opposite side. Treatments began on day 14 after transplantation. Epothilone B was administered by iv injections at a dose of 4 mg / kg, either once or once a week (days 14, 21 and 28), or every second week (days 14 and 28). 5-fluoracil was administered iv at a dose of 75 mg / kg on days 14, 21, 28. The data presented were obtained for animals surviving by day 32 (the fourth day after the last treatment). Antitumor activity was expressed as T / C% (average increase in tumor volumes in treated animals divided by the average increase in tumor volumes in control animals multiplied by 100). Tumor regression (%) was evaluated by comparing the final average tumor volume with the average tumor volume at the beginning of the experiment. Changes (Δ) in tumor volume represent tumor volume on the last day of treatment minus tumor volume on the first day of treatment.

Example 11: Phase 1 of a study to determine the dose of epothilone B, applied individually by administration once a week to adult patients with advanced dense tumors

Number of centers 2

Goals

Primary: characterization of safety profiles, including both acute and cumulative toxicity, and determination of the maximum tolerated dose of epothilone B, applied individually by intravenous infusion once a week to adult patients who have developed dense tumors that are not amenable to standard systemic therapy, or for which There is no standard systemic therapy.

Additional:

1. Determination of the pharmacokinetic characteristics of epothilone B, applied individually by intravenous infusion once a week to this group of patients; the obtained data can be used in conjunction with pharmacodynamic data (for example, with hematological parameters) to detect correlations of pharmacokinetic / pharmacodynamic parameters (PK / PD), which are required to assess safety and efficacy;

2. Obtaining preliminary evidence of the antitumor activity of epothilone B, applied individually by intravenous infusion once a week to a specified group of patients.

3. Finding correlations between drug levels in tumors in adult patients with developed dense tumors who are injected with epothilone B individually by intravenous infusion once a week, and levels whose efficacy has been established in preclinical models.

4. Obtaining information on the pharmacogenetics of tumors using tumor biopsy samples suitable and available before and after therapeutic intervention, in order to identify genes that correlate with efficacy and response; this can be established either by genetic analysis of the expression of a single gene (for example, the expression level of p53, Mar4 and mdr1 proteins), or using gene chip technology.

Description of experience

The experience was an open study using increasing doses, designed to determine the safety, pharmacokinetic and pharmacodynamic characteristics of epothilone B, administered by intravenous infusion once a week to adult patients with dense tumors that are not amenable to standard systemic therapy, or for which there is no standard systemic therapy.

The treatment period included up to 24 doses administered weekly. Treatment of patients who found an unacceptable level of toxicity or progressive development of the disease was stopped until the end of the experiment. Patients who achieved a complete or partial reaction, or patients whose disease condition stabilized after 24 doses were administered, continued further treatment according to the extension protocol as decided by the research doctor and after the sponsor’s permission. Individual patients underwent additional treatment cycles until they showed a progressive development of the disease or an unacceptable level of toxicity.

To determine the MTD, a standard phase 1 protocol was used with 3-6 patients included in the group. The dose increase was carried out according to the modified Fibonacci scheme and it was based on the toxicity levels detected for the first 4 weekly doses for each group of patients. The initial dose was 0.1 mg / m ² , the subsequent doses were as follows: 0.2, 0.3, 0.5, 0.7 and 0.9 mg / m ² .

As a preliminary value of MTD, the dose immediately preceding the dose was taken, for which dose limiting toxicity (ODT) was detected in at least two out of 3-6 patients. Then, additional patients were included in the MTD group, bringing the total number to 12, in order to confirm the MTD value based on further safety assessments, pharmacokinetic and pharmacodynamic profiles of epothilone B.

All toxicity levels were evaluated according to the revised General Toxicity Criteria of the US National Cancer Institute. The protocol determined the value of the ODT; however, as a rule, the nature of CCT is such that it is considered unacceptable even in the case of incurable solid tumors.

Patients

Inclusion criteria

To be included in the study group, the following conditions were required:

1. Age of male or female patients ≥ 18 years.

2. The presence of a dense tumor that is not amenable to standard systemic therapy and another additional systemic therapy, established on the basis of histological analysis, or for which there is no standard systemic therapy.

3. The presence of at least one measurable, measurable, or quantitatively underestimated symptom of the disease according to the criteria for the reaction of a dense tumor of the Southeast Oncological Group (SWOG), including a tumor marker value that exceeds the established normal upper limit.

4. Women who are able to have children should have a negative reaction in a pregnancy test using β-HCG serum before starting studies. Male and female patients who are capable of having children must agree to use effective contraceptives during the study period and within 3 months after stopping the medication.

5. The score (assessment) of performance according to the criteria of the World Health Organization (WHO) ≤ 2.

6. Life expectancy of at least 3 months.

7. Written consent obtained prior to any selection procedure.

Exclusion criteria

Patients were not allowed to participate in the study if at least one of the following conditions was met:

1. Female patients who are pregnant or breast-feeding. Postmenopausal women should not have menstruation for at least 12 months to confirm that they cannot have children.

2. The patient had a serious and / or untreated clinical illness (for example, uncontrolled diabetes, congestive heart failure, myocardial infarction less than 6 months before the start of the experiment, chronic kidney disease or an untreated acute infectious disease).

3. The patient had detected brain metastases.

4. The patient had an acute or detected chronic liver disease (for example, chronic active hepatitis, cirrhosis).

5. The patient had an established diagnosis of human immunodeficiency virus (HIV) infection.

6. The patient received any agent intended for the study, less than 30 days before the start of the experiment.

7. The patient underwent chemotherapy treatment less than 4 weeks before the start of the experiment (6 weeks for nitrosoureas or myomycin C).

8. The patient was treated with radiation less than 4 weeks before the start of the experiment.

9. The patient has previously undergone radiotherapy treatment involving ≥ 25% of the bone marrow.

10. The patient underwent extensive surgery less than 2 weeks before the start of the experiment.

11. There was evidence that the patient did not comply with medical regimens.

12. The patient had impaired liver, kidney, or hematological disorders characterized by the following laboratory parameters:

platelet count <100 · 10 ⁹ / l;

absolute neutrophil level (ANC) <1.5 · 10 ⁹ / l;

serum ALT (SGPT)> 2.5 × established normal upper limit (IULN);

total serum bilirubin> 1.5 × IULN;

serum creatinine> 1.5 × IULN.

13. The patient was cured of another primary malignant disease less than five years ago or, in case of non-melanomatous skin cancer and cervical carcinoma in situ, had an active stage of the disease.

Sample size

This study required approximately 40 patients.

Processing

Epothilone B was supplied in separate 2 ml glass vials as a clear, colorless concentrate for intravenous injection with a concentration of 1 mg / ml. To obtain the desired final concentration of the drug for injection, the compound was dissolved in polyethylene glycol 300 (PEG 300) and diluted with 50-100 ml of 0.9% sodium chloride for injection with a purity corresponding to USP. It was administered every 7 days as a single intravenous infusion for 30 minutes.

The initial dose level was 0.1 mg / m ² . As this dose, one third of the minimum toxic dose (MTD) was taken for dogs, which were the most sensitive species of animals on which the effect of epothilone B was studied. As indicated above, the doses were increased according to the modified Fibonacci scheme. The study included a delay in treatment, dose reduction or refusal of treatment for patients who had hematological or other toxic symptoms that are known to be caused by epothilone B. The treatment consisted of a maximum of 24 doses administered weekly if the patient did not show progressive development disease or unacceptable toxicity. After the completion of the administration of 24 doses, patients who achieved a complete or partial reaction, and patients whose disease stabilized, could continue further treatment according to the extension protocol by decision of the research doctor and after the consent of the sponsor.

Security

Indicators

The safety of epothilone B was determined on the basis of physical study and assessment of vital characteristics, results of clinical laboratory studies, side effects and taking into account the use of concomitant drugs. Side effects caused by both the drug and random factors were evaluated using the revised General Cancer Institute's Expanded General Toxicity Criteria.

Efficiency

Indicators

Although this phase 1 of the study was not intended to evaluate effectiveness, it demonstrated activity as a function of the reaction rate of the tumor being treated and the length of the period when its progressive development was absent and overall survival.

Key tumor scores included the optimal detection of all measurable, evaluated, and non-evaluated characteristics of the disease. Assessments included physical examination and a chest radiograph and, if necessary, a computerized tomogram of the chest, abdomen and pelvis; sonogram of the abdomen and pelvis; a bone scintigram in combination with a bone x-ray and the detection of all known bone lesions; and determination of tumor marker values. Detailed studies were carried out every 6 weeks and after cessation of treatment.

Objective status was evaluated in a clinical setting using Novartis guidelines based on SWOG response criteria. All full and partial cancers had to be confirmed by a second evaluation at least four weeks later. For each patient, the best tumor response was calculated using SWOG response criteria.

Pharmacokinetics

For cycles 1 and 2, the following pharmacokinetic parameters were calculated and analyzed: t _max , C _max , λ _z , t _1/2 , AUC and R _A R _A represents the ratio AUCτ _cycle2 / AUCτ _cycle1 and characterizes the accumulation coefficient. A preliminary assessment of dose proportionality is based on the AUC for the last dose among groups treated with different doses. To obtain a safety indicator, correlations of PK / PD with the observed toxicity (for example, hematopoietic) were found.

Pharmacodynamics

Before treatment and after the first cycle of therapy, samples of tumor biopsies were obtained, if this was possible and affordable. These biopsy samples were prepared for analysis of their gene expression using gene chip technology, then separately analyzed for p53 status, MAP4 DNA expression (microtubule-associated protein) and mdr1 RNA expression.

Statistical Methods

Patients with clinical side effects caused by treatment were identified (especially patients with manifestations of dose-limiting toxicity) or with deviations (arising again or increased from the initial level) of vital signs established on the basis of laboratory tests or as a result of physical studies, and such cases were quantified . The coefficient of anomalies in the group was calculated. The coefficients of the target reactions (including both complete and partial reactions) in the group were determined. Descriptive statistics were used to summarize the main pharmacokinetic parameters in the group.

Example 12: Phase 1 of the EPO906 Agent Dose Determination Study (Epothilone B). used individually by injection once every three weeks to adult patients with developed dense tumors

Number of centers 2

Location Glasgow, UK, Newcastle, UK

Goals

Basic: characterization of safety profiles, including both acute and cumulative toxicity, and determination of the maximum tolerated dose of epothilone B, applied individually by intravenous infusion once every three weeks to adult patients with developed dense tumors that are not amenable to standard systemic therapy, or for which there is no standard systemic therapy.

Additional:

1. Determination of the pharmacokinetic characteristics of epothilone B, applied individually by intravenous infusion once every three weeks to this group of patients; the data obtained can be used to identify correlations of pharmacokinetic / pharmacodynamic parameters (FC / PD), which are required to assess safety and efficacy;

2. Obtaining preliminary evidence of the antitumor activity of epothilone B, applied individually by intravenous infusion once every three weeks to a specified group of patients.

3. Finding correlations between drug levels in tumors in adult patients with developed dense tumors who are injected with epothilone B individually by intravenous infusion once every three weeks, and levels whose efficacy has been established in preclinical models.

4. Obtaining information about tumors using tumor biopsy samples suitable and measurable before and after therapeutic intervention, in order to identify biological factors that correlate with efficacy and response.

Description of experience

The experience was an open study with increasing doses, designed to determine the safety, pharmacokinetic and pharmacodynamic characteristics of epothilone B, administered by intravenous infusion once every three weeks to adult patients with dense tumors that are not amenable to standard systemic therapy, or for which there is no standard systemic therapy.

The treatment period included up to six 21-day cycles. Treatment of patients who found an unacceptable level of toxicity or progressive development of the disease was stopped until the end of the experiment. Patients who achieved a complete or partial reaction, or patients whose disease condition stabilized after six cycles, continued further treatment according to the extension protocol by decision of the research doctor and after the sponsor’s permission. Individual patients underwent additional treatment cycles until they showed a progressive development of the disease or an unacceptable level of toxicity.

In the absence of dose-limiting toxicity (ODT), the dose was increased as follows:

1. The first dose increase: a dose increase of 100% (until in the first group toxicity of degree 2 was detected, in this case, the dose increase was 25-67%).

2. Increasing the dose when moving from the first group to the second group after a 100% increase in dose: increase in dose by 67% until toxicity of degree 2 is detected.

3. Recent dose increases after detecting degree 2 toxicity: dose increases of 25-67% as agreed between the researchers and the sponsor.

The dose increase was based on the toxicity levels obtained in the first cycle of studies for each group of patients. As a preliminary value of MTD, we took the dose level immediately preceding the level for which dose limiting toxicity (ODT) was detected in at least two out of 3-6 patients. Then, additional patients were included in the preliminary MTD group, bringing the total number to 12 in order to confirm the MTD value based on further safety assessments, pharmacokinetic and pharmacodynamic profiles of epothilone B.

An increase in dose for the same patient was not allowed.

All toxicity levels were evaluated according to the National Cancer Institute's Revised Toxicity Criteria. The protocol determined the value of the ODT; however, as a rule, the ODT value is such that it is considered unacceptable even in the case of incurable solid tumors.

Patients

Inclusion criteria

To be included in the study group, it was necessary to satisfy the following criteria:

1. Age of male or female patients ≥ 18 years.

2. The presence of a dense tumor that is not amenable to standard systemic therapy, and another additional systemic therapy, or for which there is no standard systemic therapy, established based on histological analysis.

3. The presence of at least one measurable, assessed, or underestimated symptom of the disease, determined according to the criteria for the reaction of a dense tumor of the South-Eastern Oncological Group (SWOG), including a tumor marker value that exceeds the established normal upper limit.

4. Women who are able to have children should have a negative reaction in a pregnancy test using P-HCG serum before starting the tests. Male and female patients who are capable of having children must agree to use effective contraceptives during the study period and within 3 months after stopping the medication.

5. The health score according to the criteria of the World Health Organization (WHO) ≤ 2.

6. Life expectancy of at least 3 months.

7. Written consent obtained prior to any selection procedure.

Exclusion criteria

Patients were not allowed to participate in the study if at least one of the following conditions was met:

1. Female patients who are pregnant or breast-feeding. Postmenopausal women should not have menstruation for at least 12 months to confirm that they cannot have children.

2. The patient had a serious and / or untreated clinical disease (for example, uncontrolled diabetes, congestive heart failure, myocardial infarction less than 6 months before the start of the experiment, chronic kidney disease, or an untreated acute infectious disease).

3. It was established that the patient had metastases in the brain.

4. The patient had an acute or detected chronic liver disease (for example, chronic active hepatitis, cirrhosis).

5. The patient had an established diagnosis of human immunodeficiency virus (HIV) infection.

6. The patient received any agent intended for the study, less than 30 days before the start of the experiment.

7. The patient underwent chemotherapy treatment less than 4 weeks before the start of the experiment (6 weeks for nitrosoureas or myomycin C).

8. The patient was treated with radiation less than 4 weeks before the start of the experiment.

9. The patient has previously undergone radiotherapy treatment involving ≥ 25% of the bone marrow.

10. The patient underwent extensive surgery less than 2 weeks before the start of the experiment.

11. There was evidence that the patient did not comply with medical regimens.

12. The patient had impaired liver, kidney, or hematological disorders characterized by the following laboratory parameters:

platelet count <100 · 10 ⁹ / l;

absolute neutrophil level (ANC) <1.5 · 10 ⁹ / l;

serum ALT (SGPT) or ASR (SGOT)> 2.5 × established normal upper limit (IULN) (> 5 × IULN for patients with liver metastases);

total serum bilirubin> 1.5 × IULN;

serum creatinine> 1.5 × IULN.

13. The patient was cured of another primary malignant disease less than five years ago or, in case of non-melanomatous skin cancer and cervical carcinoma in situ, had an active stage of the disease.

Sample size

This study required approximately 40 patients.

Processing

Epothilone B was supplied in separate 2 ml glass vials as a clear, colorless concentrate for intravenous injection with a concentration of 1 mg / ml. To obtain the desired final concentration of the drug for injection, the compound was dissolved in polyethylene glycol 300 (PEG 300) and diluted with 50 or 100 ml of 0.9% sodium chloride for injection with a purity corresponding to USP. It was administered every 21 days for six cycles in the form of single intravenous infusions lasting 30 minutes.

The initial dose level was 0.3 mg / m ² . As this dose, one third of the minimum toxic dose (MTD) was taken, which was established for dogs, which were the most sensitive species of animals, on which the effect of epothilone B was studied. Since using the smaller of the 2 doses that were administered to dogs in a GLP toxicological study, since the dose was 0.1 mg / kg with a single repeat after 3 weeks, there were no deaths, the MTD value was taken to be approximately 0.05 mg / kg. Using a coefficient of 20 to convert the dose for dogs, measured in mg / kg, to the dose for people, measured in mg / m ² , the initial dose was obtained according to the following formula: 1/3 · 0.05 mg / kg · 20 kg / m ² = 0.3 mg / m ² .

The increase in doses was carried out according to the scheme described above.

The study included treatment delays, dose reduction or refusal of treatment for patients who had hematological or other toxic symptoms, which are known to be caused by epothilone B. Treatment was carried out for a maximum of 6 cycles if the patient did not detect progressive development of the disease or unacceptable toxicity. After completion of 6 cycles, patients in whom a complete or partial reaction was achieved, and patients in whom the disease stabilized, could continue further treatment according to the extension protocol by decision of the research doctor and after the consent of the sponsor.

Safety indicators

The safety of epothilone B was determined on the basis of physical study and assessment of vital characteristics, results of clinical laboratory studies, side effects and taking into account the use of concomitant drugs. Side effects caused by both the drug and random factors were evaluated using the revised General Cancer Institute Toxicity Criteria.

Performance indicators

Although this phase 1 of the study was not intended to evaluate effectiveness, it demonstrated activity as a function of the reaction rate of the tumor being treated and the length of the period when its progressive development was absent and overall survival. Key tumor scores included the optimal detection of all measurable, evaluated, and quantitatively underestimated characteristics of the disease. Assessments included physical examination and a chest radiograph and, if necessary, a computerized tomogram of the chest, abdomen and pelvis; sonogram of the abdomen and pelvis; a bone scintigram in combination with a bone x-ray and the detection of all known bone lesions; and determination of tumor marker values. Detailed studies were carried out every two cycles and after termination of treatment.

Objective status was evaluated in a clinical setting using Novartis guidelines based on SWOG response criteria. All complete and partial reactions should have been confirmed by a second evaluation at least four weeks later. For each patient, the best tumor response was calculated using SWOG response criteria.

Pharmacokinetics

For cycles 1 and 2, the following pharmacokinetic parameters were calculated and analyzed: t _max , C _max , λ _z , t _1/2 , AUC and R _A. R _A represents the ratio AUCτ _cycle2 / AUCτ _cycle1 and characterizes the accumulation coefficient. A preliminary assessment of dose proportionality is based on the AUC for the last dose among groups treated with different doses. To obtain a safety indicator, correlations of PK / PD with the observed toxicity (for example, hematopoietic) were found.

Pharmacodynamics

To identify biological factors correlating with efficacy and response, tumor biopsy samples were obtained before therapy and after the first cycle of therapy, if this was possible and affordable.

Statistical Methods

Patients with clinical side effects caused by treatment (especially patients with dose-limiting toxicity) or with abnormalities (arising again or increasing from the initial level) of vital signs identified on the basis of laboratory tests or as a result of physical studies were identified, and such cases were quantified. The coefficient of anomalies in the group was calculated. The coefficients of the target reactions (including both complete and partial reactions) in the group were determined. Descriptive statistics were used to summarize the main pharmacokinetic parameters in the group.

The discussion of the results:

In general, the results presented in the examples indicate that treatment with epothilone B is effective:

a) in relation to a tumor that does not respond to standard treatment, for example, a colon tumor that does not respond to treatment with 5-fluoracil, or which does not respond to treatment using TAXOL®;

b) also in relation to a tumor that cannot be treated using TAXOL®, for example lung cancer, especially non-small cell lung cancer, and / or epidermoid tumors, especially cervical tumors;

c) also in relation to orthotopic tumors and the formation of metastases, for example, in prostate tumors;

d) also in relation to breast cancer, where in vitro analyzes (example 3) showed a higher activity of epothilone B compared with the activity of TAXOL®.

Preferred drug regimens range from weekly treatment in doses of about 1/3 to 2/3 of the MTD to a single dose treatment up to the MTD, with the best treatment regimen corresponding to the administration of the drug from once a week to once every three weeks.

Claims (29)

1. A method of treating tumor diseases, including the introduction of epothilone B, characterized in that epothilone B is administered with an interval of one to six weeks in a dose that is calculated for people according to the formula single dose (mg / m ² ) = (from 0.1 to y) × N, where Y = 6; N is the number of weeks between administration. 2. The method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered more than once with an interval of one to six weeks between administrations. 3. A method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered to adults in a dose that is calculated by the formula (II) single dose (mg / m ² ) = (from 0.1 to 2.5) × N, where N has the above meaning. 4. A method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered weekly at a dose of from 0.1 to 6 mg / m ² . 5. The method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered every 6-8 days at a dose of 0.1 to 5 mg / m ² . 6. The method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered every 6-8 days at a dose of 0.1 to 3 mg / m ² . 7. A method for the treatment of tumor diseases according to claim 1, characterized in that epothilone B is administered weekly at a dose of 0.3 to 1 mg / m ² . 8. The method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered every third week at a dose of 0.3 to 18 mg / m ² . 9. A method for treating tumor diseases according to claim 1, characterized in that epothilone B is administered once every three weeks, i.e. every 18-24 days, at a dose of 0.3 to 12 mg / m ² . 10. The method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered every 18-24 days at a dose of 0.3 to 7.5 mg / m ² . 11. A method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered every third week at a dose of 1.0 to 3.0 mg / m ² . 12. The method of treating tumor diseases according to claim 1, characterized in that epothilone B is administered by intravenous infusion. 13. A method of treating tumor diseases according to claim 4, characterized in that epothilone B is administered by intravenous infusion. 14. A method of treating tumor diseases according to claim 8, characterized in that epothilone B is administered by intravenous infusion. 15. A method for the treatment of tumor diseases according to item 13 or 14, characterized in that each infusion is carried out for 5-30 minutes 16. A method of treating tumor diseases according to one of claims 1 to 14, characterized in that the tumor disease cannot be treated with one or more chemotherapeutic agents other than epothilone, where epothilone, especially epothilone B, is administered to a person in need of such treatment, in a dose acceptable for the treatment of this disease. 17. A method for treating tumor diseases according to claim 16, wherein the non-treatable tumor is selected from the group consisting of a lung tumor, a colorectal tumor, a prostate, breast, or epidermoid tumor of the head or neck. 18. The method of treating tumor diseases according to clause 16, wherein the tumor to be treated is a colorectal tumor that cannot be treated with at least one anti-cancer agent from the taxane class. 19. The method of treating tumor diseases according to claim 18, characterized in that the colorectal tumor to be treated is also not treatable by at least one other standard chemotherapeutic agent. 20. A method for the treatment of tumor diseases according to clause 16, characterized in that the tumor to be treated is a prostate tumor and / or any metastases thereof that cannot be treated with hormones. 21. The method of treating tumor diseases according to clause 16, characterized in that the tumor to be treated is an epidermoid tumor of the head or neck that cannot be treated with at least one other chemotherapeutic agent. 22. The method of treating tumor diseases according to clause 16, wherein the tumor to be treated is a lung tumor that cannot be treated with at least one other chemotherapeutic agent. 23. A method for treating tumor diseases according to claim 16, characterized in that the tumor to be treated is a breast tumor. 24. A method for treating tumor diseases according to claim 16, characterized in that the tumor to be treated is a colorectal tumor that does not respond to standard chemotherapy. 25. A method of treating tumor diseases according to any one of claims 1 to 15, characterized in that the tumor disease to be treated is selected from the group consisting of a colorectal tumor, a genitourinary tumor, an epidermoid tumor, a lung tumor and a breast tumor. 26. A method for treating tumor diseases according to any one of claims 1 to 16, characterized in that the tumor disease to be treated is a tumor having multiple drug resistance. 27. A method of treating tumor diseases according to any one of claims 1 to 16, characterized in that the tumor disease to be treated is selected from the group comprising melanoma, ovarian cancer, pancreatic cancer, neuroblastoma, head or neck cancer, bladder cancer, kidney cancer , brain cancer and stomach cancer. 28. The method of treating a tumor disease according to claim 16, characterized in that epothilone B is administered to a person at a dose of 1 to 100% of the maximum tolerated dose (MTD), and in the form of one or more additional doses, each of which is in the range from 1 to 100% of the MTD and which are then administered in the form of at least one additional treatment with intervals between treatments of one to three weeks. 29. A method of treating a tumor disease according to any one of claims 1 to 28, further comprising the step of administering (a) epothilone B in combination with (b) another antitumor therapeutic agent, wherein the combination treatment regimen is structured so that component (a) and component (b) was administered to a person in need of such treatment, in combination with each other and in amounts that together are therapeutically effective against a tumorous disease.