Hydrated crystalline camptothecin esters for treating cancer

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention describes a crystalline hydrate of camptothecin esters, namely the crystalline hydrates of camptothecin aliphatic esters, a pharmaceutical composition containing the crystalline hydrates of camptothecin aliphatic esters for treating cancer or malignant tumours, as well as a method for treating cancer or malignant tumour.

EFFECT: what is prepared and described is the new crystalline form of the hydrate of camptothecin aliphatic ester possessing low toxicity and good absorbability in a living body.

19 cl, 5 ex, 10 tbl, 5 dwg

 

The technical FIELD TO WHICH the INVENTION RELATES.

The present application claims the benefit in accordance with §35 U.S.C. 120 on the basis of the provisional application for U.S. patent No. 11/923727, filed October 25, 2007, which is fully included in the description of the present application by reference.

The present invention relates to esters of camptothecin and the use of esters of camptothecin for the treatment of cancer, tumors and the like.

The LEVEL of TECHNOLOGY

Toxicity (or side effects)suffered by patients as a result of taking anti-cancer drugs taken during chemotherapy remains the biggest problem associated with cancer treatment. therapeutic index (TI) anti-cancer agent is defined as the ratio of the most acceptable dose to effective dose. It is known that most anticancer agents clinically used at present, has a very narrow range of T (1 to 1.2). In other words, the toxic effects of most anti-cancer agents in patients receiving chemotherapy, which is inevitable when the levels of the effective dose due to narrow therapeutic index associated with anticancer agents. Differences in toxicity between agents are degrees of seriousness, and types of violations. Fluorouracil 5-FU) is one of the most commonly used chemotherapeutic agents for the system and palliative treatment of patients with cancer formations, what is happening in the gastrointestinal tract, breast, head and neck. Digidropirimidindegidrogenazy (DPD) is the initial and limiting the speed of the enzyme in the catabolism of 5-FU. The DPD deficiency is increasingly recognized as an important pharmacogenetic violation in the etiology of serious toxicity associated with 5-FU. Reported that cancer patients who were genetically heterozygous or homozygous for mutant alleles of the gene encoding DPD, suffered from serious toxicity, including death, after administration of 5-FU. Van Kuilenburg ABP, Haasjes J, Richel DJ, et al. Clinical implications of dihydropyrimidinedehydrogenase (DPD) deficiency = MKD in patients with severe 5-fluorouracil-associated toxicity: identification of new mutations in the DPD gene. Clinical Cancer Res 2000, 6: 4705-4712. Van Kuilenburg ABP, Muller EW, Haasjes J et al. Lethal outcome of a complete dihydroprimidine dehydrogenase (DPD) deficiency = MKD after administration of 5-florouracil: frequency of the common IVS14+1G>A mutation causing DPD deficiency = MKD. Clin Cancer Res 2001, 7: 1149-1153. Platinum agents are also commonly used to treat. An extensive literature has confirmed side effects of platinum compounds. Cisplatin has numerous toxic effects: nephrotoxicity, neurotoxicity, ototoxicity, nausea and vomiting. DeVita VT, Hellman S, and Rosenberg SA. Cancer, Principles and Practice of Oncology, 7th Ed, Lippincott Williams & Wilkins 2005, 335. Nephrotoxicity of cisplatin almost led to its abandonment, prior to the introduction of potent hydration of Cvetkovi and his staff that predated is whether the development of acute renal failure. Cvitkovic E, Spaulding J, V Bethune, et al. Improvement of cis-dichlorodiamineplatinum (NSC 119875): therapeutic index in an animal model. Cancer 1977, 39, 1357. Hayes D, Cvitkovic E, Golbey R, et al. High dose cis-platinum diamine dichloride: amelioration of renal toxicity by mamnitol diuresis. Cancer 1977, 39, 1372. Believed that the toxicity of cisplatin was some force in history as when searching for less toxic analogues, and for more effective treatment of its side effects. Myelosuppression, which usually was not caused by a severe action of cisplatin, has doselimiting toxicity carboplatin. Evens B, Raju K. Calvert A, et al. Phase II study of JM8, a new platinum analogue, in advanced ovarian carcinoma. Cancer Treat. Rep., 67, 997, 1983. Doselimiting toxicity oxaliplatin is a sensory neuropathy - a characteristic of all DACH-containing derivatives of platinum. DeVita VT, Hellman S, and Rosenberg SA. Cancer, Principles and Practice of Oncology, 7th Ed, Lippincott Williams & Wilkins 2005, 335. Alkylating agents also play an important role in the treatment of cancer. Each alkylating agent is associated with a specific toxicity and are not discussed individually. The following types of toxicity are typical alkylating agents: hematopoietic toxicity, gastrointestinal toxicity, gonadotoxicity, pulmonary toxicity, alopecia, teratogenicity, carcinogenesis and immunosuppression. DeVita VT, Hellman S, and Rosenberg SA. Cancer, Principles and Practice of Oncology, 7th Ed, Lippincott Williams & Wilkins 2005, 335. Normal doselimiting toxicosis alkylating agent is a hematopoietic toxicity. Agents that interact with topoisomerase, increasingly attract the attention of clinical oncologists for their unique mechanism of action. Topotecan is a semisynthetic product of 10-hydroxycamptothecin of natural origin and is indicated for use in the treatment of second-line progressive resistant ovarian cancer and small-cell lung cancer, and he was also active in the treatment of hematological malignancies, including myelodysplastic syndromes and multiple myeloma. Huinink W, Gore M, Carmichael J, et al. Topotecan versus paclitaxol for the treatment of recurrent epithelial ovarian cancer. J Clin Oncol 1997, 15, 2183; Schiller JH, Adak S, Cella D, et al. Topotecan versus observation after cisplatin plus etoposide in extensive-stage small-cell lung cancer: E7593-a phase II trial od the Eastern Cooperative Oncology Group. J Clin Oncol 2001, 19, 2114; von Pawel J, Schiller JH, Shepherd FA, et al. Topotecan versus cyclophosphamide, doxorubicin, and vincristine for the treatment of recurrent small-cell lung cancer. J Clin Oncol 1999, 17, 658, Pizzolato JF, Saltz LB. The camptothecins. Lancet 2003, 361, 2235. Doselimiting toxicity of this agent represents the myelosuppression. Although topotecan was combined with various other treatments, including ionizing radiation, cisplatin, paclitaxel and doxorubicin in clinical trials, none of these combinations has not reached any of the standard applications in clinical Oncology. This may be partly due to the multiple mielosupressivei toxicity of topotecan, which makes it difficult merge the use of high doses of other agents, suppressive bone marrow. Miller AA, Lilenbaum RC, Lynch TJ, et al. Treatment-related fatal sepsis from topotecan/cisplatin and topotecan/paclitaxol. J Clin Oncol 1996, 14, 1964. Irinotecan is also a semi-synthetic product group camptothecin. This connection is indicated for use as a single agent or in combination with fluorouracil and leucovorin in the treatment of patients with colorectal cancer formations and also found active in the treatment of small-cell lung cancer when introduced in combination with cisplatin. This combination was also active in the treatment of non-small cell lung cancer. Saltz LB, Cox JV, Blanke C, et al. Irinotecan plus florouracil and leucovorin for metastatic colorectal cancer. Irinotecan study group. N Engl J Med 2002, 343, 905; Douillard JY, Cunningham D, Roth AD, et al. Irinotecan combined with florouracil compared with florouracil alone as first-line treatment for metastatic colorectal cancer: a multicenter randomized trial. Lancet 2000, 355, 1041; Pizzolato JF, Saltz LB. The camptothecins. Lancet 2003, 361, 2235. Doselimiting toxicity of irinotecan are neutropenia and diarrhea, delayed-type, and thus, its application in clinical Oncology is also limited. All other variety of anticancer agents, including the recently introduced Erbitux and Avastin, can be applied for a limited number of specific treatments that are also associated with toxicity. Thus, for cancer researchers and clinical oncologists still face a difficult task in finding the best and the clients with a wider therapeutic index for the treatment.

It was shown that camptothecin, a cytotoxic alkaloid originally isolated from the leaves and bark of Camptotheca Acuminata (Nyssaceae) Voll and his colleagues (J. Am. Chem. Soc. 88, 3888, 1966), has antitumor activity against system leukemia mice L 1210. Structure camptothecin, alkaloid, which has a standard occurring group of indole alkaloid (Heckendorf et al., J Org. Chem. 41, 2045, 1976), shown below in formula (X).

This connection ("CPT") has Pyh ring system with only one asymmetric center in the ring E with 20(S)-configuration. Pyh ring system is part of pyrrolo[3,4-b]quinoline (rings A, B and C), conjugated pyridone (ring D) and six-membered lactone (ring E) with an α-hydroxyl group. Camptothecin was a great interest from the time of its initial allocation due to its noteworthy activity against system leukemia mice L 1210. Earlier data on the antitumor activity of camptothecin were obtained using experimentally transplanted malignant tumors such as leukemia L-1210 in mice or tumor Walker 256 in rats (Chem. Rev. 23, 385, 1973, Cancer Treat. Rep. 60, 1007, 1967). Subsequent clinical studies have shown that this compound is not suitable for use as an anticancer agent in vivo AFL is stvie its high toxicity. Camptothecin insoluble in water. Consequently, earlier camptothecin were evaluated clinically as a water-soluble salt of carboxylate sodium. This form of camptothecin had strong toxicity and, apparently, there had been no anti-cancer activity (Gottlieb et al., Cancer Chemother. Rep. 54, 461, 1970, and 56, 103, 1972, Muggia et al., Cancer Chemother. Rep. 56, 515, 1972, Moertel et al., Cancer Chemother. Rep. 56, 95, 1972, and Schaeppi et al., Cancer Chemother. Rep. 5:25, 1974). These results caused the termination of the test phase II. Continued evaluation of this agent showed that the salt of the carboxylate sodium is only 10% of such active as native camptothecin closed lactone ring in an unmodified form (Wall et al., In International Symposium on Biochemistry And discrimination of The Alkaloids, Mothes et al., eds., Academie - Verlag, Berlin, 77, 1969, Giovanella et al., Cancer res. 51, 3052, 1991). In addition, we established the important indicators of antitumor activity in a group of camptothecin (Wall et al., Ann. Rev., Pharmacol. Toxicol. 17, 117, 1977). These results show that the lactone ring E unchanged and α-hydroxyl group is essential for antitumor activity.

In the mid 1980's it was discovered that the molecular target of action of camptothecin represents a new nuclear enzyme topoisomerase I. Hsiang YH, Liu LF. Identification of mammalian DNA topoisomerase I as an intracellular target of the anticancer drug camptothecin. Cancer Res1988, 48, 1722. At about the same time received and biologically octilinear new water-soluble derivatives of camptothecin, including two connections (topotecan and irinotecan), discussed earlier. Subsequent clinical evaluation of two compounds showed predictable toxicity and significant anticancer activity. Takimoto CH, Arbuck SG. Topoisomerase I targeting agents: the camptothecins. In: Chabner BA, Longo DL, eds., Cancer therapy & biotherapy: principles and practice, 3rd ed. Philadelphia: Lippincott Williams & Wilkins 2001, 579. Topotecan was approved in 1996 as the drugs of the second line for the treatment of progressive ovarian cancer, and he later received recognition for the treatment of patients with refractory small cell lung cancer. At the same time irinotecan was registered for the treatment of progressive 5-fu-resistant colorectal cancer. He was actually the first new agent that has received approval for the treatment of this disease in the United States through almost 40 years.

In 1989 Giovanella et al. found that some of the water-insoluble derivatives of camptothecin have high antitumor activity against xenografts of human tumors (Giovanella et al., Science, 246, 1046, 1989). It was also shown that the introduction of camptothecin closed lactone ring is superior to the introduction of a water-soluble salt of carboxylate (Giovanella et al., Cancer Res., 51, 3052, 1991). These findings were additionally confirmed the importance of the lactone ring in an unaltered view of the biological activity.

The ring opening 20(S)-camptothecin leads to a much stronger anticancer activity in mice than in humans. In fact, it was proved that CPT, given intramuscularly ("/m"), subcutaneously ("p/C") and inside the abdomen ("their recommendations carry considerable."), is a very strong anticancer agent against human tumors in mice, that is, when growth as xenografts in Nude mice (Giovanella et al., Cancer Res. 51:3052, 1991). However, when the tumors were treated with CPT in humans, was shown a lower degree of anticancer activity in humans than in mice (Stehlin et al., In Camptothecins: New Anticancer Agents, 1995, CRC Press, pp. 59-65).

The same phenomenon was observed with other derivatives of CPT. It was shown that mice 9-nitrocamptothecin ("9NC) is 2-3 times more potent than CPT in respect of xenografts of human tumor, causing the complete destruction of all human malignant tumors that are treated (Pantazis et al., Cancer Res. 53:1577, 1993; Pantazis et al., Int. J. Cancer 53:863, 1995).

The ring opening is particularly problematic in camptothecin that exist in two different forms. A naturally occurring camptothecin has the S-configuration, and 10-100 times more biologically active than the R-isomer. I believe that the lactone form S-configuration required for antitumor activity, and the form of the carboxylate usually refers to clinical toxicities. M is lekule is in equilibrium in aqueous solution. This equilibrium depends on the pH. At physiological pH, that is 7 or higher, the equilibrium equation is as follows:

The hydrolysis reaction of the biological active lactone ring camptothecin with water at higher pH gives biologically inactive open form. In addition, the problem of hydrolysis with CPT and its analogs exacerbated in human blood, because the predominant human serum albumin (HSA) is preferably associated with the form of carboxylate, which shifts the equilibrium of the lactone/carboxylate to an inactive form (J. Biochem., 212, 285-287, 1993; Biochemistry, 33, 10325-10336, 1994; Biochemistry, 33, 12540-12545, 19944). Accordingly, the preservation of the lactone ring of the molecule in sufficient time for the tumor cells to cyclically pass through S-phase, is a major challenge, was the object of a considerable number of studies.

Many attempts were made to obtain the derivative camptothecin possessing higher biological activity and higher stability. Many of these compounds are products of modifications A, B and C rings of the molecule, but few of these modifications increased the stability of the lactone ring under physiological conditions. Other approaches have been more successful. For example, acylation of the group of 20-HE provides a useful item for which the ITA lactone ring E/ Wall et al., U.S. patent No. 4943579 describes several acylated compounds of camptothecin having solubility in water, although the lactone may not remain intact under physiological conditions. U.S. patent No. 5968943 Cao et al. reveals CPT derivatives, which are effective anticancer agents.

In the literature was informed about the many different reactions obtain the esters of camptothecin.

Direct acylation of camptothecin anhydrides of organic acids in the presence of pyridine as a catalyst used for obtaining esters of alkyl - and alkenylbenzene (as shown above). This reaction usually produces a high output, but the availability of anhydrides of organic acids limits the possibility of reaction.

Therefore, used a system of reagents dicyclohexylcarbodiimide (DCC)/dimethylaminopyridine (DMAP) to acylation reactions of carboxylic acids with alcohols and thiols. Previously, the method used for the preparation of aromatic esters of camptothecin (as shown below).

This technique, however, gives a good outputs reactions only when carboxylic acids are very electrophilic. When acids are less electrophilic reaction gives out low or no floor shall indicate the expected product. For example, using propionic acid to produce propionate of camptothecin according to this method, the ester product is essentially not received and the source camptothecin returned almost 100%.

Nonane chloride is also used as Alliluyeva agent for the esterification of camptothecin in the presence of pyridine as an agent, catching HCl in methylene chloride. The reaction (as shown below) was held from low yield (6%).

Since the original structural features of camptothecin provide cancer researchers ideal basis for structure modification in obtaining the best anti-cancer agents, synthesized and evaluated a variety of different esters of camptothecin. It was found that the products of esterification significantly increased the biological life of the molecules in the body. It was also shown that treatment of synthetic esters of camptothecin of human tumors grown in xenografts in Nude mice, is effective, and toxicity in mice is minimal. Cao, Z.; Pantazis, P.; Mendoza, J.; Early, J.; Kozielski, A.; Harris, N.; Vardeman, D.; Liehr, J.; Stehlin, J.; Giovanella, B. Ann. N.Y. Acad. Sci. 2000, 922, 122; Cao, Z.; Pantazis, P.; Mendoza, J.; Early, J.; Kozielski, A.; Harris, N.; Giovanella, B. Acta Pharmacologica Sinica 2003, 24, 109.

While there are many ways to obtain esters of camptothecin, however, each method has the certain restrictions, as discussed previously. Therefore, there is still a need in the opening esters of camptothecin, who will retain antitumor activity of the parent compound, CPT, will have much lower toxicity than CPT and which can be produced in accordance with the alternative method(s) of receipt esters of camptothecin.

The INVENTION

Accordingly, a feature of the present invention is to provide a crystalline hydrate of ester of camptothecin, which will demonstrate a broad spectrum of antitumor activity and preferably low or non-observable toxicity.

Another feature of the present invention is to provide a crystalline hydrate of ester of camptothecin with a wider range of therapeutic index than most anticancer agents currently used.

Another feature of the present invention is to provide a safe and simple method of obtaining a crystalline hydrate of ester of camptothecin.

A further feature of the present invention is to provide crystalline hydrates of ester of camptothecin that preserve significant antitumor activity as the original connection, CPT, and have amnog the lower toxicity, than its original connection.

Another feature of the present invention is to provide crystalline hydrates of ester of camptothecin with good absorption capacity in a living organism.

To achieve the features and in accordance with the purposes of the present invention, as implemented and widely discussed in the present description, the present invention is due in part to the crystalline hydrates of ester of camptothecin, such as crystalline hydrates complex aliphatic ester camptothecin as a crystalline hydrate camptothecin-20-O-propionate ("CZ48"), pharmaceutical compositions comprising crystalline hydrates aliphatic ether complex, methods for treating cancer or a malignant tumor, using crystalline hydrates of ester of camptothecin, and methods of producing hydrated crystalline esters of camptothecin, including the interaction of compounds camptothecin at least one agent allermuir that is protonated at least one acid.

Additional features and advantages of the present invention will be formulated in part in the description that follows, and will partly be obvious from the description or may be learned by practice of the present invention. Tasks and other to becoming the VA of the present invention will be apparent and attained by means of the elements and combinations in particular mentioned in the description and the attached claims.

All patents, patent applications and publications referred to throughout the application, are incorporated fully by reference into the present description and are part of this application.

It should be understood that both the foregoing General description and the following detailed description are only exemplary and explanatory and are intended to provide further explanation of the present invention in the claims.

BRIEF DESCRIPTION of DRAWINGS

Figure 1 is an ORTEP diagram of single molecules of product.

Figure 2 is an ORTEP diagram depicting two molecules of crystalline hydrate camptothecin-20-O-propionate, United together water bridge.

Figure 3 is an ORTEP diagram depicting the three-dimensional structure (a, b position coordinates) of the crystalline hydrate camptothecin-20-O-propionate.

Figure 4 is an ORTEP diagram depicting the three-dimensional structure (a, c position coordinates) of the crystalline hydrate camptothecin-20-O-propionate.

Figure 5 is an ORTEP diagram depicting the three-dimensional structure (b, c position coordinates) of the crystalline hydrate camptothecin-20-O-propionate.

DETAILED DESCRIPTION of the PRESENT INVENTION

Camptothecine ("CPT") oblad who have significant antitumor and anticancer activity but these connections are subject to destruction under normal physiological conditions, and the resulting metabolites often exhibit toxic properties. Conducted laboratory studies of the metabolism of camptothecin in plasma showed that the only identified metabolite is a salt of carboxylate sodium with open ring, which is toxic and inactive. Measurement of pharmacokinetics for CPT in human plasma indicates that the half-life of the medicinal product with the lactone ring intact represents approximately 30 minutes These results imply that the drug loses 90% of its activity, and there are many toxicities in a very short time after its acceptance by the patients.

According to the present invention disclosed CPT transformed into more laboratorial and gidratirovannuyu crystal molecule of ester in the future from time to time called prodrug. When receiving patients prodrug is rapidly injected into the bloodstream of patients and are easily converted into the original connection in the body. Noted that the prodrug may also be active and/or may be partially or completely converted.

According to different variants of implementation of the compounds of the present invention may include crystallites is their aliphatic esters CPT in hydrated form, having the formula:

In this formula, n may represent any number in the range from 1 to 10 or more, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. R2represents a C2-C6alkyl group, such as C2-C4alkyl group. In one or more embodiments, the implementation of R2represents-CH2CH3; -CH2CH2CH3; -CH2CH2CH2CH3; or-CH2CH2CH2CH2CH2CH3. As another specific implementation, n is 3 and R2represents-CH2CH3or-CH2CH2CH3.

According to at least one variant of implementation of the compounds of the present invention may include a crystalline hydrate camptothecin-20-O-propionate having the formula shown below:

In this formula, n may represent any number in the range from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

As an example of the structure of the crystalline hydrate camptothecin-20-O-propionate was determined using x-ray analysis of the single crystal and is shown in Fig.1-5. Figure 1 shows the ORTEP diagram of single molecules of product. One molecule of camptothecin-20-O-propionate and about 3 water molecules are connected by h is through strong hydrogen bonds. More than one molecule of camptothecin-20-O-propionate in the crystal structure can be connected to each other through the bridge, created from molecules H2O. Figure 2 shows two molecules camptothecin-20-O-propionate, United water bridge. Fig.3-5 shows the three-dimensional crystal structure. Other esters CPT of the present invention will have the same structure except for the chain length of ester.

Prodrugs of the present invention can be in liquid or solid state. According to the present invention, the compounds are crystalline. According to at least some variants of implementation, the compounds of the present invention can be crystalline or solid in which the constituent atoms molecules or ions are Packed in a regularly ordered, repeating pattern, located in all three spatial directions. Crystals can be monoclinically system sizes (from 0.10 to 0.50)×(0.01 to 0.10)×(0.01 to 0.05) mm3and volumes from 100 to 5000 Å3such amounts as from 200 to 4500 Å3or from 500 to 4000 Å3or from 750 to 3500 Å3or from 1000 to 3000 Å3or from 1250 to 2500 Å3or from 1500 to 2200 Å3.

Used in this description of "crystalline" refers to a material that contains certain compounds is s, which can be gidratirovannym and/or solvated and has sufficient content of crystals to show pronounced diffraction model using powder x-ray diffraction (XRPD) or other diffraction methods. Often crystalline material, which is obtained from the solvent by direct crystallization of the substance dissolved in the solution, or interconversion of crystals obtained under different crystallization conditions, will have crystals that contain a solvent, called the crystalline MES. Examples of properties of crystals include chemical orientation of the parts relative to each other within the crystal and the prevalence of specific forms of compounds which is preferred in the presence of acid in the composition of the solvent.

According to at least one variant of implementation, the compounds of the present invention may have a purity of from about 90% to about 100% by AUC (area under the curve). According to some variants of implementation, the compounds of the present invention may have a purity of from about 95% to about 100% for AUC. According to at least one variant of implementation, the compounds of the present invention may have a purity of from about 99% to about 100% by AUC, such as from 99,3% to 99,999%; there are tons of 99.5% to 99.999%availability; from of 99.75% to 99,999%; 99,85% to 99,999% of all AUC. According to at least one variant of implementation, the compounds of the present invention can have a melting point of approximately 240°C to approximately 243°C, such as 242°C or about 242°C. This point and melting point is preferable for the crystalline hydrate of ester, where R2represents-CH2CH3. The melting point of the compounds of the present invention can be lower or higher than the above range, when R2represents a group CH3or C3H7or C4H9or C6H13.

According to different variants of implementation, the present invention may include crystalline hydrates aliphatic esters CRT with S-configuration, R-configuration and/or racemic mixture of both S - and R-isomers. According to some variants of implementation, crystalline hydrates aliphatic esters PTD derived from natural camptothecin, only have the S-configuration or mainly S-configuration, such as 90% or higher, 95% or higher, 98% or higher, 99% or higher, or from 99% to 99.99%.

The conversion of the prodrugs in the original connection, CPT, may be mediated by a group of enzymes, called esterases present in the blood of many animals, including humans. Since Pro is carstvo quickly distributed throughout the body within a short period of time after delivery, these compounds exist at very low concentrations in a time when they are subjected to enzymatic hydrolysis, which released the original camptothecin, which prevents the deposition of CPT in the bloodstream.

Therefore, the present invention provides a hydrated crystalline CPT derivatives or analogs which are preferably longer remain unchanged in the body of a person or animal, especially in the human body, thereby enhancing anti-tumor and anti-cancer effects without producing unwanted side effects.

The present invention also provides pharmaceutical compositions containing a pharmaceutically effective amount of one or more compounds of the present invention optionally in combination with one or more pharmaceutically acceptable carriers, excipients, diluents or excipients. For example, the compounds of the present invention can be obtained in the form of tablets, pills, powder mixtures, capsules, injectable formulations, solutions, suppositories, emulsions, dispersions, additives in food and in other suitable forms. They can also be obtained in the form of sterile solid compositions, for example, dried by sublimation and, if desirable, combined with other pharmaceutically pickup is acceptable-excipients. Such solid compositions may be dissolved in sterile water, physiological saline solution or a mixture of water and an organic solvent, such as propylene glycol, ethanol and the like, or some other sterile enter the environment directly before the use of parenteral administration.

Typical pharmaceutically acceptable carriers are, for example, mannitol, urea, dextrans, lactose, non sugar, potato and maize starches, magnesium stearate, talc, vegetable oils, polyalkylene glycols, ethylcellulose, poly(vinyl pyrrolidone), calcium carbonate, etiloleat, isopropylmyristate, benzyl benzoate, sodium carbonate, gelatin, potassium carbonate, silicic acid. Pharmaceutical drug can also contain non-toxic auxiliary substances such as emulsifying, preserving and/or wetting agents such as, for example, sorbitan monolaurate, the triethanolamine oleate, polyoxyethylene the monostearate, glyceryltrinitrate, dioctyl sodium sulfosuccinate and the like.

"Pharmaceutical composition" refers to mixtures of one or more of the compounds described in this application, or its pharmaceutically acceptable salts with other chemical components such as pharmaceutically acceptable carriers and/or excipients. CE is ü pharmaceutical composition is to facilitate the introduction of compounds into the body.

Used in the present description, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or environment for medicines, such as liquid or solid filler, diluent, excipient, solvent or encapsulating material, included in the transfer or transportation of the test agent from one organ or body part to another body or body part. Each carrier must be "acceptable" in the sense of being compatible with other ingredients of the composition and is not harmful for the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) powder tragakant; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and waxes suppositories; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as etiloleat and tillaart; (13) agar; (14) buffering agents, such as hydroxide magnesium and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical formulations. Physiologically acceptable carrier should not cause significant irritation to an organism and does not abolish the biological activity and properties of the compound that is administered.

"Excipient" can refer to an inert substance added to a pharmaceutical composition to further facilitate the introduction of the connection. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, fats, lipids, vegetable oils and polyethylene glycols.

"Pharmaceutically effective amount" may also or alternatively indicate the amount that is able to provide therapeutic and/or prophylactic effect. Dosage is introduced compounds according to this invention to obtain therapeutic and/or prophylactic effects will, of course, identified with specific details related to this case, including, for example, some entered the compound, route of administration, the condition being treated, and the person being treated. Typical egene the Naya dose (introduced in single or divided doses) will contain a dose level of from about 0.01 mg/kg to about 50-100 mg/kg of body weight of active compound of the invention. The daily dose can be from about 0.05 mg/kg to about 20 mg/kg, or from about 0.1 mg/kg to about 10 mg/kg

In some embodiments, implementation of the preferred therapeutic effect is, to some extent, inhibition of growth characteristic of cell proliferative diseases such as breast cancer or pancreatic cancer. therapeutic effect is also usually, but not in case of need, will weaken to some extent one or more symptoms, in addition to cell growth or size of the cell mass. therapeutic effect may include, for example, one or more of 1) reducing the number of cells; 2) reduction of the size of the cells; 3) inhibition (that is, slowing to some extent, preferably stopping) cell infiltration into peripheral organs, for example, in the case of metastasis of cancer; 3) inhibition (that is, slowing to some extent, preferably stopping) of tumor metastasis; 4) inhibition, to some extent, the growth of the cells; and/or 5) weakening to some extent one or more symptoms associated with the disorder.

The pharmaceutical compositions used in the methods of the present invention may also contain the active ingredient in a form suitable for oral administration, for example, as a tablet and, mints, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use can be obtained according to any method known in the art for pharmaceutical compositions, and such compositions may contain sweeteners, flavors, coloring agents and/or preservatives. Tablets may contain the active ingredient in a mixture with non-toxic pharmaceutically acceptable excipients which are suitable for receiving tablets. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and dezinfeciruyuhimi agents such as microcrystalline cellulose, croscarmellose sodium, corn starch, or alginic acid; binding agents, e.g. starch, gelatin, polyvinylpyrrolidone or gum, and lubricating agents, e.g. magnesium stearate, stearic acid or talc. Tablets may be uncoated or coated by known techniques to hide the taste of the medicinal product or to delay the disintegration and absorption in the gastrointestinal tract and thereby prolong the automatic action over a longer period. Can be used suitably, for example, water-soluble material that hides the taste, such as hypromellose or hydroxypropylcellulose, or a material with a retention time, such as ethylcellulose or acetate butyrate cellulose.

Formulations for oral use may also be presented as hard gelatin capsules where the active ingredient is mixed with an inert solid diluent, for example calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water-soluble carrier such as polyethylene glycol or an oil medium, for example peanut oil, liquid paraffin or olive oil.

Aqueous suspensions may contain the active material in a mixture with excipients suitable for receiving water suspensions. Such excipients can be suspendresume agents, for example, carboxymethylcellulose sodium, methylcellulose, hypromellose, sodium alginate, polyvinylpyrrolidone, tragakant and the Arabian gum; dispersing or wetting agents may be a phosphatides of natural origin, for example, lecithin, or condensation products of accelerated with fatty acids, for example, a polyoxyethylene stearate, and the condensation products of ethylene oxide with aliphatic alcohols with long chain for example, heptadecadiene-axiatonal, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from anhydrides of fatty acids and hexitol, for example, polyethylene, sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl-p-hydroxybenzoate, one or more dyes, one or more flavoring agents, and one or more sweeteners, such as sucrose, saccharin or aspartame.

Oil suspensions can be obtained by suspendirovanie active ingredient in a vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or in mineral oil such as liquid kerosene. Oily suspensions may contain a thickening agent, for example, yellow beeswax, hard paraffin or cetyl alcohol. Sweeteners such as defined above, and flavoring agents may be added to provide palatable oral drug. These compositions may be preserved by the addition of an antioxidant, such as bottled hydroxyanisol or α-tocopherol.

Dispersible powders and granules suitable for gaining the aqueous slurry by adding water, provide the active ingredient in a mixture with dispersing or wetting agent, suspenders agent and one or more preservatives. Examples of suitable dispersing or wetting agents and suspendida agents are already mentioned above. May also contain additional excipients, for example, sweeteners, flavorings and colorings. These compositions may be preserved by adding an antioxidant such as ascorbic acid.

Compounds and pharmaceutical compositions used in the methods of the present invention may also be in the form of emulsions of oil-in-water". The oil phase may be a vegetable oil, such as olive oil or peanut oil, or mineral oil, for example liquid paraffin or mixtures thereof. Suitable emulsifying agents may be phosphatides of natural origin, for example, soybean lecithin, and esters or partial esters derived from anhydrides of fatty acids and hexitol, for example, sorbitan monooleate, and condensation products of the above-mentioned incomplete complex ester with ethylene oxide, e.g. polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners, flavorings, preservatives and/or antioxidants.

Syrups and elixirs can be obtained with padsl what stitely, for example, glycerin, propylene glycol, sorbitol or sucrose. Such compositions may also contain softeners, preservatives, fragrances, colorants and antioxidants.

The pharmaceutical compositions can be in the form of sterile injectable aqueous solutions. Among the acceptable fillers and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution.

Sterile injectable drug may also be a sterile injectable microemulsion "oil-in-water", in which the active ingredient is dissolved in the oil phase. For example, the active ingredient may be first dissolved in a mixture of soybean oil and lecithin. The oil solution is then injected into the mixture of water and glycerol and treated with obtaining a microemulsion.

The pharmaceutical compositions can be in the form of a sterile injectable suspension in an aqueous or oil-based for intramuscular and subcutaneous injection. This suspension can be obtained according to known in the art methods using suitable dispersing or wetting agents and suspendresume agents which have been mentioned above. Sterile injectable drug may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable the second diluent or solvent, for example, as a solution in 1,3-butanediol. In addition, sterile fatty oils normally used as a solvent or suspendida environment. This purpose can be applied to any soft fatty oils, including synthetic mono - or diglycerides. In addition, fatty acids such as oleic acid find use in obtaining injectable drugs.

Compounds of the present invention used in the methods of the present invention can also be entered in the form of suppositories for rectal administration of medicinal means. These compositions can be obtained by mixing the inhibitor with a suitable not cause irritation excipient, which is solid at ordinary temperatures but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, contains glycerin gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycol of various molecular masses and esters fatty acids of polyethylene glycol.

For local application can be used in creams, ointments, jellies containing drug solutions or suspensions, etc. containing the compound or composition of the invention. Used in this description of the local skin is tion may include a liquid for rinsing the mouth and throat.

The compounds used in the methods of the present invention, it is possible to enter in intranasal form via topical application of suitable intranasal fillers and delivery systems, or via transdermal routes of administration, using the forms transdermal patch, is well known to specialists in this field of technology. For administration in the form of a transdermal delivery system, the introduction of the dosage, of course, be continuous rather than intermittent throughout the dosage regimen.

The methods, compounds and compositions of the present invention can also be used in conjunction with other known therapeutic agents that are selected for their particular usefulness against the condition that is being treated. For example, these compounds can be used in conjunction with known anti-cancer and cytotoxic agents. Additionally, the present methods and compounds can also be used in conjunction with other inhibitors of the parts of the signaling pathway that links the receptor growth factor cell surface nuclear signals that trigger cell proliferation.

One aspect of the invention is a pharmaceutical composition suitable for treating cancer in warm-blooded animals, which contains at least one compound of the invention as defined in n the standing description together with pharmaceutically acceptable excipients. Composition was prepared in accordance with known methods of obtaining compositions suitable for oral, local, transdermal, rectally, via inhalation, parenteral (intravenous, intramuscular or intraperitoneal) administration, and the like. Detailed methodological guidance for obtaining the compositions of the invention found through links on the 18th or 19th edition Remington's Pharmaceutical. Sciences, Published by the Mack Publishing Co., Easton, Pa. 18040. The relevant parts are included in the present description by reference.

Considered forms of single doses or multiple doses, each having advantages in certain clinical conditions. A single dose contains a predetermined quantity of active compound calculated to obtain the desired effect(s) in the treatment of cancer. Form multiple doses may be particularly useful when the number of individual doses or fractional doses are required to achieve the desired effects. Both of these dosage forms can have the description of the invention, they are caused or directly depend on the characteristics of a particular compound, a particular therapeutic effect to be achieved, and any limitations inherent in getting a certain connection to lacenaire.

The unit dose may contain a therapeutically effective amount sufficient to treat cancer of the object, and may contain from about 1.0 to 1000 mg of the compounds, for example, from about 50 to 500 mg.

The compound may be administered orally in a suitable solvent, such as a tablet for swallowing, buccal tablet, capsule, dripping, elixir, suspension, syrup, toffee, capsule, tablet for sucking and the like. Typically, the most effective composition is a tablet or capsule (separately or together defined as "oral unit dosage"). Suitable compositions receive in accordance with standard available methods of composition, which correspond to the characteristics of the connection to the excipients that are available to obtain the composition. Tablet or capsule will contain from about 50 to about 500 mg of the compounds of the present invention.

The form can deliver a fast connection or may be a drug with a slow release. The connection can be placed in a hard or soft capsule, may be compressed into tablets, or may be incorporated in a beverage, food or other food. The percentage of the final compositions and medicaments may, of course, be varied and can mill artno be between 1 and 90% of the weight of the final form, for example, tablets. The number in such therapeutically effective compositions represents such, that you will get the right dosage. Compositions according to the present invention can be obtained so that the oral form of unit dosage contains from about 5 to about 50 percent weight (% wt.) unit dosing by weight between 50 and 1000 mg

A suitable composition for oral unit dosage may contain: a binder, such as tragakant, gum, corn starch, gelatin; sweeteners, such as lactose or sucrose; dezintegriruetsja agents such as corn starch, alginic acid and the like; lubricants such as magnesium stearate; or flavoring agents such as peppermint, oil of Grushenka or similar. Various other materials may be present as coatings or to otherwise modify the physical form of the oral unit dosage. Oral unit dosage may be coated with shellac, sugar or both substances. A syrup or elixir may contain the compound, sucrose as a sweetener, methyl - and propylparaben as preservatives, a dye and flavoring. Any material used must be pharmaceutically acceptable and substantially non-toxic. Details of the types of prednimustine can be found in the nineteenth edition of "Remington: The Science and Practice of Pharmacy", Mack Printing Company, Easton, Pa. For a more complete discussion, see especially chapters 91-93.

Connection, you can enter parenterally, for example intravenously, intramuscularly, subcutaneously or intraperitoneally. Media, or excipient, or a mixture of excipients can be a solvent or dispersion medium containing, for example, various polar or nonpolar solvents, suitable mixture or oil. Used in this description of "the media" or "excipient" means a pharmaceutically acceptable carrier or excipient and includes any and all solvents, dispersion agents or the environment, covering(I), antimicrobial agents, ISO/Hypo/hypertensive agents that alter the absorption agents and the like. The use of such substances and agents for pharmaceutically active substances is well known in the art. The use in therapeutic compositions are considering except when any standard medium or agent is incompatible with the active ingredient. In addition, other or additional active ingredients may also be included in the final composition.

Solutions the compound can be obtained in suitable diluents such as water, ethanol, glycerol, liquid polyethylene glycol(and), various oils and/or their mixtures and other substances known to experts in the Anna field of technology.

The pharmaceutical forms suitable for using injection include sterile solutions, dispersions, emulsions, and sterile powders. The final form should be stable under conditions of manufacture and storage. In addition, the final pharmaceutical form must be protected from contamination and, therefore, must be able to inhibit the growth of microorganisms, such as bacteria or fungi. Can be entered as a single intravenous or intraperitoneal dose. Alternatively, it may be used to slow long-term infusion or repeated short-term daily infusion, usually lasting from 1 to 8 days. You can also apply through the day, or dosing once every few days.

Sterile injectable solutions are inclusion compounds in the required amount in one or more appropriate solvents, to which if necessary can be added other ingredients mentioned above or known to specialists in this field of technology. Sterile injectable solutions are inclusion compounds in the required amount in an appropriate solvent, if necessary, with various other ingredients. Followed by sterilization techniques, such as filtering. Typically, the dispersion is obtained by switching the connection of sterilized the hydrated filler, which also contains a dispersion medium and the required other ingredients as shown above. In the case of sterile powder to which you add any ingredients, preferred methods include vacuum drying or freeze drying.

It is noted that in all cases the final form must be sterile and should easily pass through the device for injection, such as a hollow needle. Appropriate viscosity can be achieved and maintained, by a suitable choice of solvents or excipients. In addition, it may be pianino the use of molecular or particulate coating such as lecithin, proper selection of particle size in the dispersions or the use of materials with properties of surface-active substances.

Prevention or inhibition of growth of microorganisms can be achieved by adding one or more antibacterial agents, such as chlorobutanol, ascorbic acid, parabens, thimerosal or similar. May also preferably be the inclusion of agents that modify toychest, such as sugar or salt.

In some cases, for example, where the compound of the invention represents a completely insoluble in water, it may be appropriate to provide a liposomal delivery. The system limits with the unity of invention by the introduction, by encapsulation, environment or inclusion compounds of the invention in, on or near the lipid vesicles, or liposomes, or micelles.

In one or more embodiments of implementation of the present invention the present invention relates to methods for treating various forms of cancer, tumors and/or precancerous conditions or precursors of malignant tumors. The method may include the treatment of cancer or malignant tumor in a patient by introducing the compound or the composition containing the compound of the present invention. The method may include the introduction of an effective amount of the compound or the composition containing the compound of the present invention, for treating cancer or a malignant tumor, in which the cancer or malignant tumor amenable to treatment with a compound or composition comprising the compound of the present invention. Below is additionally described that different cancers and malignant tumors can be treated using the present invention. Compounds of the present invention is effective in the treatment of human or animal cancers, malignant tumors, neoplasm or precancerous lesions. Specific examples include, but are not limited to, leukemia, melanoma, liver cancer, breast cancer, colorectal, rectal, Yai is nick, prostate, stomach, bladder, desmoplastic small-round-cell tumor (DSRCT), cancer of the pancreas, lungs, kidneys, colon, tumors of the Central nervous system or any combination. Used in the present description, the term "malignant tumor" is intended to encompass all forms of human carcinoma or animals, sarcomas and melanomas that occur in poorly differentiated, moderately differentiated and well-differentiated forms.

Another important feature of the compounds of the present invention relates to a relatively low or no evident overall toxicity of prodrugs, which is administered in accordance with the idea of the present description. General toxicity can be assessed using different criteria. For example, loss of body weight of the object more than 10% of the originally registered body weight (i.e. before treatment) can be regarded as one of the signs of toxicity. In addition, as evidence of toxicity can also be interpreted by the loss of total mobility and activity and signs of diarrhea or cystitis object.

Crystalline hydrates aliphatic esters CPT demonstrate a wide spectrum of activity without significant toxicity in mice at various dose ranges. therapeutic index can be defined the n through testing on naked mice such as the average therapeutic index of tests on mice with xenograft tumors, which is a tumor of the bladder, breast, colon, kidney, lung, melanoma, pancreas, prostate, ovary and/or any of the cancers mentioned in the present description. Additionally, therapeutic index of this agent significantly improved compared with most anticancer agents clinically used at present by the oncologists. therapeutic index of the crystalline hydrates of aliphatic esters CPT may be in the range from 2 to 500 (for example, from 3 to 500, from 4 to 50, from 3 to 10, from 4 to 15, 5 to 20, from 8 to 20, 10 to 20, 25 to 500, 50 to 500, from 75 to 500, 100 to 500, from 150 to 500, 200 to 500, 250 to 500, 300 to 500, from 350 to 500, from 400 to 500, 450 to 500), thus believe that 2000 mg/kg represents the most acceptable dose. therapeutic index for most anticancer agents currently used in clinical Oncology, however, represents approximately 1, which is very narrow. Additionally, none of the currently used anticancer agents cannot be continuously used over a long period of effective dose. Compounds of the present invention can complianc is but to use every day or weekly, or monthly for 2 months, from 3 months to 12 months from 4 months to 15 months, from 5 months to 15 months, from 6 months up to 24 months or more.

According to different variants of implementation, the hydrates are crystalline aliphatic esters CPT can be introduced together with pharmaceutically acceptable carriers or diluents. For example, such pharmaceutical compositions may normally contain, for example, pharmaceutically acceptable salts, buffering agents, preservatives and/or compatible media. As used in the present description, a "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers, diluents or encapsulating substances which are suitable for administration to mammals, including humans. Pharmaceutically acceptable carrier may be, for example, one or more gelatin capsules, cholesterol pellets microsuspension in lipid and lepidoptery emulsions (Intralipid 10, Intralipid 20 or natural oils) or other suitable emulsifiers for lipophilic compounds. The amount of active ingredient (crystalline hydrates aliphatic esters CPT)contained in the pharmaceutical compositions according to the invention can vary depending on many is their factors, such as route of administration and types of targets (e.g., types of cancer), compounds that use.

When treating or delaying the growth of malignant tumors in mammals, in accordance with the present invention is administered compounds or pharmaceutical compositions of the present invention using methods known to experts in the art, such as intramuscular, intravenous, transdermal or oral administration. Usually you can apply known methods, for example, gelatin capsules for oral administration, as well as the compositions, such as microsuspension in lipid and lepidoptery emulsions (e.g., Intralipid 20, cottonseed oil and peanut oil) for intramuscular and inclusion in cholesterol pellets for subcutaneous long-term administration. Another way of introducing the compounds of the present invention is a transdermal or percutaneous path. One example of such implementation is the use of the patch. In particular, the patch can be obtained with highly dispersed suspension of the compounds disclosed in this application, for example, dimethylsulfoxide (DMSO) or a mixture of DMSO with cotton oil, and put in contact with skin tumor of a mammal from the location of the tumor inside the skin bag. Other media Il is their mixtures with other solvents and solid carriers would work equally well. The patch may contain hydrate CPT of the present invention in the form of a solution or suspension. The patch can then be applied to the skin of the patient, for example, by inserting it into a skin bag of the patient, which is formed by folding and stitching the skin together by means of stitches, clips or other retaining devices. This bag should be used in such a way to ensure continuous contact with the skin without the intervention of a mammal. In addition to using skin bag can be applied to any device which ensures a stable placement of the patch in contact with the skin. For example, you can use adhesive tape to hold the patch in the appropriate place on the skin.

Used in this description of an "effective amount" of the compounds of the present invention is intended to denote the number of compounds that will inhibit the growth, or slow down the cancer, or to destroy malignant cells and cause the regression and the weakening of malignant tumors, i.e. to reduce the volume or size of such tumors or eliminate the tumor completely.

Effective amounts can be introduced mammals, including humans, based on the surface area of the body. The interdependence of dosage changes for animals of various sizes and species and the La people (based on mg/m 2body surface) is described E.J. Freireich et al., Cancer Chemother. Rep., 50(4): 219 (1966). The surface area may be approximately determined from the height and weight of the object (see, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y. pp.537-538 (1970)). An effective amount of compounds of camptothecin in the present invention can range from about 10 to about 1000 mg/m2body surface per day.

Preferred effective amount or dosage of the compounds or prodrugs of the present invention are from about 1 to about 100 mg of prodrug/kg body weight twice a week with the intramuscular route of administration, and from about 1 to about 500 mg of prodrug/kg/day by oral route of administration. An effective amount or dosage of CPT derivatives or prodrugs of the present invention are, for example, from about 1 mg/kg/week to about 100 mg/kg/week prodrugs with transdermal route of administration. For all paths entering precise timing of the introduction of dosages can be varied to achieve optimal results. Typically, when using Intralipid 20 as a carrier for prodrugs actual dosage prodrugs of reaching the patient will be less. This is due to some loss Proletarsk is on the walls of the syringe, needles and containers drugs, which is common in case of suspension of Intralipid 20. When using media, such as cottonseed oil, above this loss is not so much felt, because the prodrug so strongly adheres to the surface of syringes, needles, containers of medicines and the like. For example, preferably, it was found that, as a rule, approximately 2.5 mg of prodrug/kg of body weight two times a week, using cottonseed oil, which is administered by intramuscular, will deliver the same amount to the patient as 4.0 mg of prodrug/kg of body weight two times a week, using Intralipid 20 as a carrier. Usually from about 1 mg to about 4 mg of the prodrug is added to from about 0.1 ml to about 1 ml of media.

In one or more embodiments of implementation of the present invention, one or more compounds of the present invention (or a composition comprising the compound(I) of the present invention can be soluble in the liposomes. Liposomes can include, for example, lipids such as cholesterol, phospholipids, or micelles, consisting of surfactants, such as, for example, sodium dodecylsulfate, octylphenoxypolyethoxyethanol or sorbitan monooleate. As a rule, prodrugs contact the lipid Balaam membrane liposomes with high affinity. A prodrug that is associated with the liposome, can preferably be inserted between the acyl chains of the lipid. The lactone ring derived camptothecin, prodrug, associated with the membrane, thus removed from the aquatic environment inside and outside the liposomes and thus protected from hydrolysis. Because the drug is associated with the liposome, protected from hydrolysis, the antitumor activity of the drug is maintained. For prodrugs of camptothecin, which have a lower affinity with the membrane of the liposome and thus separated from the membrane of a liposome that is inside the liposomes, the pH of the internal parts of liposomes can be reduced, thereby preventing hydrolysis of the prodrugs of camptothecin.

Group liposomal delivery systems, which can be used in accordance with the present invention, includes described in U.S. patent No. 5552156 and 5736156, which are included in the present description fully by reference. Other liposomal delivery system that can be used in accordance with the present invention include liposomes containing active agents, United with lipids or surfactants as described in U.S. patent No. 5827533 and 5882679; lipid vesicles formed from salts of fatty acids of alkylamine, as described in point is the awning U.S. No. 5874105; liposomes to encapsulate the dry powder compositions of the active agent, as described in U.S. patent No. 5783211; liposomal delivery system medicinal products for the local patches, as described in U.S. patent No. 5718914; liposomes are described in U.S. patent No. 5631237; composition of liposomes and lipid complexes described in U.S. patent No. 5549910 and 5077057; liposomes used for sustained release of steroid drugs, as described in U.S. patent No. 5043165; liposomes are described in U.S. patent No. 5013556; and liposomes are described in U.S. patent No. 4663161; all of which are included in the present description in its entirety by reference.

Single-layer liposomes, also called single-layer vesicles, are spherical vesicles consisting of one membrane lipid bilayer, which defines a closed space. The membrane bilayer is composed of two layers of lipids; the inner layer and the outer layer. The outer layer of lipid molecules oriented portions of their hydrophilic heads into the external aqueous environment and the hydrophobic tails in the interior of the liposomes. The inner layer of the lipid lies directly below the outer layer; the lipids are oriented with their heads in the water of the inner part of the liposomes and their tails to tails of the outer layer of the lipid.

Multilayer liposomes, also the so-called multilayer vesicles, composed of more than one lipid bilayer membrane, the membrane of which is determined by more than one confined space. Membranes are concentrically arranged so that different membrane was separated by space like onion peel.

Used in the present description, the phrase "liposomal prodrug" denotes that or some portion or all of the prodrug of camptothecin are located in one or more parts of the liposomes or micelles, or prodrug of camptothecin associated with the membrane of liposomes. The delivery system can be a liposome, comprising the membrane lipid bilayer that surrounds the prodrug of camptothecin. Used in the present description, the phrase "associated with a lipid membrane" means that at least lactone ring some or all of the prodrugs of camptothecin associated with the lipid membrane of the liposome and the liposome contains more than one membrane bilayer, the prodrug of camptothecin associated with at least 1 membrane. Those prodrugs of camptothecin that have a high affinity to the membrane, tend to remain associated with the membrane. Prodrugs of camptothecin with low affinity to the membrane of the liposomes is at least partly separated from the membranes of liposomes and stay in the space of the liposomes.

In the us oasam description defined that micelles are spherical, consisting of a monolayer of the membrane, which defines a closed space, and the membrane is composed of molecules of surface-active substances, oriented such that the hydrocarbon tails are directed to a closed space, and the polar heads oriented to the external aqueous environment. Prodrugs of camptothecin when binding to micelles are or space associated with the membrane, micelle, or associated with the outer surface of the micelle.

Liposomes were successfully used for administering drugs to cancer patients, and have shown that they are clinically appropriate for delivery of anti-cancer drugs such as doxorubicin, daunorubicin complexes and cisplatin. Forssen, et al., Cancer Res. 1992, 52: 3255-3261; Perez-Soler, et al. Cancer Res. 1990, 50: 4260-4266; and Khokhar, et al. J. Med. Chem. 1991, 34: 325-329, all of which are included in the present description in its entirety by reference.

Similarly micelles also used for delivery of medicines to patients (Brodin et al., Acta Pharm. Suec. 19 267-284 (1982)) and micelles have been used as carriers of drugs for targeted drug delivery (D. D. Lasic, Nature 335: 279-280 (1992); and Supersaxo et al., Pharm. Res. 8: 1286-1291 (1991)), including anticancer drugs (Fung et al., Biomater. Artif. Cells. Artif. Organs 16: 439 et. seq. (198); and Yokoyama et al., Cancer Res. 51: 3229-3236 (1991)), all of which are included in the present description in its entirety by reference.

Liposomes and/or micelles of the present invention containing prodrugs of camptothecin, can be entered cancer patient usually intravenously. Liposomes and/or micelles rush circulatory system to cancer cells, where membrane vesicles connected with the membrane of a cancer cell, thus releasing prodrug of camptothecin in a cancer cell, or where the liposomes and/or micelles remain adjacent to cancer cells, the prodrug of camptothecin diffuses from the liposomes and/or micelles to get into the cancer cells.

Any lipid or mixture of lipids, which form liposomes and/or micelles, are suitable for use in the present invention. Suitable are phosphatidylcholine, including, for example, L-α-dimyristoylphosphatidylcholine (DMPC), L-α-dipalmitoylphosphatidylcholine (DPPC) and L-α-distearoylphosphatidylcholine (DSPC). Also suitable are phosphatidylglycerol, including, for example, L-α-dimyristoylphosphatidylcholine (DMPG). DMPC and DMPG, both are in the liquid phase at 37°C, while DSPC is in solid phase at 37°C. Given that the presence of negatively charged lipid in the membrane of liposomes causes the liposomes to push off from each other, a small number of such to the to, for example, approximately 10%, the negatively charged lipid, such as distearoylphosphatidylglycerol (DSPG), can be included in the liposomes of DSPC. Other suitable phospholipids include phosphatidylethanolamine, synthesised and phosphatidic acid containing lauric, myristic, palmitic, hexadecanoyl, stearic, oleic, linoleic, arachidonic, beenbuy and lignocellulose acid. Other suitable lipid comprises cholesterol.

Liposomes and/or micelles can be covered with a polyethylene glycol or protein GM1that helps the particles to avoid the reticuloendothelial system.

DSPC, since it is in the solid phase at 37°C (average temperature)limits the spread of drug camptothecin from liposomes and, thus, can be used for slow release prodrugs of camptothecin.

DMPG, DPPC and DSPC could be obtained from Avanti Polar Lipids, Alabaster, Ala. and used without additional purification. All other chemical compounds can be chemically pure and used without further purification.

Any surfactant, or mixtures thereof, which forms micelles, is suitable for use in the present invention. Suitable surfactants include sodium dodecylsulfate (LTOs), available from Kodak, Rochester,N.Y., octylphenoxypolyethoxyethanol available under the trademark "Triton X-100" from Aldrich Chemical Co., Milwaukee, Wis., and sorbitan monooleate available under the trademark "Polysorbate 80 and tween 80 from Sigma Chemical Co. Other suitable surfactants include, for example, sodium salt deoxycholic acid, sodium salt holeva acid and polyoxyethylene-10-cetyl ether, available under the trademark "BRIJ-56"; these surfactants are available from Sigma Chemical Co.

In addition, the micelles can be composed of a lipid such as a phospholipid, and mixtures of lipids. Also micelles can be composed of a lipid and a surfactant.

Suspension of liposomes can be obtained using the method of Burke and Tritton Biochemistry 24: 1768-1776 (1985), which is included in the present description in its entirety by reference. Liposomes are preferably small single-layer vesicles (SUV), not multilayer vesicles (MLV). However, as SUV and MLV are within the scope of the invention. While MLV have the advantage of speed limits diffusion linked prodrugs of camptothecin, they have the disadvantage that more easily removed by macrophages than SUV. Can be used suspensions of a mixture of lipids containing 200 mg/ml lipid in phosphate-saline buffer solution (PBS)containing 8 mm Na2HPO4, 1 mm KH2 PO4, 137 mm NaCl and 3 mm KCl, pH 7.4 and received using vortex mixing for 5-10 minutes above the phase transition temperature of the gel - liquid crystal TMlipid. The lipid suspension is then treated with ultrasound for 3-4 hours using an ultrasonic bath from Laboratory Supplies Co., Hicksville, N.Y., until they are optically pure. The decrease in pH from 7.4 to 6.8 can be followed to obtain SUV medicines DMPG; therefore, the pH of these suspensions SUV preferably set to 7.4, using a small amount of 2.5 M NaOH in PBS, and again treated with ultrasound. Each type of suspension of liposomes is preferably incubated for 30 minutes at 37°C.

Many of liposomes and micelles are described in U.S. patent No. 5552156, 7244449 and 5736156, which are included in the present description in its entirety by reference.

"Prodrugs" can be derived compounds obtained by adding the group, which provides greater solubility of the compound desired for delivery. Being in the body, the prodrug enzyme normally acts, for example, esterase, amidase or phosphatase, to obtain active compounds.

The term "alkyl", alone or in combination, refers to an optionally substituted straight-chain, optionally substituted branched-chain or optionally substituted cyclic the sky the alkyl radical, having from 1 to about 6 carbons, more preferably 2-4 carbon. Examples of alkyl radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, tert-amyl, pentyl, hexyl and the like. The term "cycloalkyl" covers a circular configuration, includes definitions of alkyl and mainly refers to monocyclic, bicyclic, tricyclic or more polycyclic alkyl radicals, in which each circular part has from 3 to about 6 carbon atoms. Examples cycloalkyl radicals include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

"Effective amount" or "effective dose" can refer to the amount necessary or sufficient for inhibiting undesirable cell growth, for example, prevent undesirable cell growth or reduce the existing growth of cells, such as tumor cell growth. The effective amount can vary depending on factors known to the person skilled in the technical field, such as the type of cell growth, the method and mode of administration, the size of the object, the intensity of cell growth and the like. Specialist in the art will be able to consider these factors and to determine the effective amount. This can be achieved by using real what about the inventions.

"Therapeutically effective antineoplastic therapy" can refer to therapy, which is effective to maintain or reduce the size of, for example, the volume of the primary tumor or metastatic tumor. This can be achieved using the present invention.

According to one or more variants of implementation of the present invention provide methods for creating crystalline hydrates complex aliphatic ester camptothecin, which include the initial step of the reaction compounds camptothecin at least one allermuir agent, protonated at least one acid, such as sulfuric acid. Allerease agent may contain a group of ester for the formation of the source camptothecin. The obtained ester of camptothecin is a crystalline hydrate complex aliphatic ether CPT. In at least one embodiment, the obtained ester of camptothecin is a crystalline hydrate camptothecin-20-O-propionate (hydrate CZ48). According to at least one variant of implementation, the original connection of camptothecin is a CPT.

The number of initial connections camptothecin may be any desired number, as there is a sufficient allerease agent accepts the stop at least part of the source camptothecin in the ester of camptothecin, as described in this application. For example, the number of source connections camptothecin can be from about 1 g to 100 g or more.

As for Alliluyeva agent, allerease agent usually in one or more embodiments, the implementation contains a group of ester to obtain the original camptothecin. Allerease agent may be derived from organic acids, such as acid anhydride. For example, allerease agent may have the formula (R1CO)2O, in which R1represents an organic group, and the group typically R1is a group that is an integral part of the accession of ester to the source connection of camptothecin. In more detail, for example, only for the purposes of the invention, the group R1can be alkyl group, such as C1-C6an alkyl group. Specific examples of the groups R1include, but are not limited to, -CH2CH3; -CH2CH2CH3; -CH2CH2CH2CH3; or-CH2CH2CH2CH2CH2CH3.

The number Alliluyeva agent used in the reaction of the present invention, may be sufficient so that the group of ester was formed on the source connection of camptothecin. The appropriate number Alliluyeva Agen is and include, but not limited to, from about 10 ml to about 1 l, on the basis of from 20 g to 30 g of the original camptothecin. The examples below provide typical number Alliluyeva agent, which can be used in the reaction.

As to the acid used in the reaction of the present invention, the acid may be used in catalytic quantities so that the acylation of the original camptothecin could be derived from organic acids, such as acid anhydride. The acid may be sulfuric acid or other acids such as HCl, HNO3or HClO4. The acid can be concentrated acid, such as concentrated sulphuric acid. The acid may be any molar concentration, such as from about 0.0001 to about 0.02 M or higher. The amount of acid used in the reaction, may be a catalytic amount, such as from about 0.1 ml to about 1.0 ml, and more preferably, from about 0,20 ml to about 0.75 ml, or about 0.5 ml of the reaction with from 20 g to 30 g of the original camptothecin. The amount of acid used to catalyze the esterification reaction may be different depending on the weight of the substances involved in the reaction.

In the present invention, according to one or more variants of implementation is tvline, various reagents can be combined together in any order or sequentially, at the same time or in any combination. Can be applied to any reaction capacity. The reaction can be carried out at any temperature above the freezing point of reagents, such as from about 20°C or higher. The reaction can be carried out at ambient temperatures or elevated temperatures, such as from about 20°C to about 110°C or higher. The reaction can be carried out without spending a lot of time, for example, from 1 minute to 1 hour or more. The reaction time depends on the amount of reagent used and the desired amount of conversion of the original CPT in the ester of camptothecin. The reaction can be carried out in an inert atmosphere or in air. Examples of the inert atmosphere may be a nitrogen atmosphere or argon atmosphere.

According to different variants of implementation, the yield of the reaction can be at least 95% to 100% weight. the source connection of camptothecin, which in turn ester of camptothecin. In at least one embodiment, the yield of the reaction represents from about 97% to 100% conversion of compounds of camptothecin in the ester of camptothecin.

The purity and/or concentration of the original CPT or reagent is insignificant. Different purity and various con is entrale can affect the percentage yield of esters, CPT, which are formed during the reaction. Preferably, the purity of the source reagent CPT represents from about 30% to about 100%. More preferably, the purity represents from about 80% to about 100% or from 90% to 99.9% or higher. Preferably, the number of reagents CPT or CPT derived represents from approximately 0.1 to approximately 50% of the total volume of the reactants. More preferably, the amount is from about 0.5 to about 5.0% of the total volume of the reactants.

pH, concentration and purity of the acid are not of great importance, since impurities in the acid does not interact with CPT or allermuir agent. The acidity of the acid must be strong enough to protonation Alliluyeva agent involved in the reaction. This ability to have a strong inorganic acid, such as H2SO4, HCl, HNO3and HClO4. For this type of catalytic esterification reaction can be used by other acids, such as AlCl3and BF3. the pH of the acid may be from about 0.5 to about 5. Preferably, the acid is concentrated and high purity. For example, the concentration may be from about 60% to about 100%. Preferably, the concentration represents from approximately the nutrient 95% to about 98%. The purity of the acid may be from about 30% to about 100%. Preferably, the purity represents from about 90% to about 100%. Preferably, the amount of acid such as concentrated sulfuric acid represents from about 0.1 to about 10% of the total volume of the reactants. More preferably, the amount is from about 0.5% to about 8.5% of the total volume of the reactants.

Preferably, the acid is added to a mixture of CPT and anhydride acid while the mixture is stirred. Preferably, the amount of acid that can be added to the mixture, is sufficient for the action of the acid as a catalyst. Preferably, from about 4 to about 8 drops of acid from glass pipettes can be added to approximately 70-100 ml acylhomoserine (such as the amount of acid can be used when allerease agent differs from acylhomoserine). However, in case of necessity, more or less acid may be added to the mixture CPT and acid anhydride, preferably while the mixture is stirred.

The mixture CPT, acid anhydride and acid can be placed in the reactor, which preferably contains an inert atmosphere, such as the N2and can be heated from approximately 0°to approximately 120°C. Preferably, the mixture is heated from about 90°C to about 110°C, and more preferably, the reactor is heated to approximately 100°C.

Preferably, the reaction to proceed until, until the desired product is formed. The reaction time can be from as short as a few hours, before long, as a few days. Preferably, the reaction time may be about 15 hours under an inert atmosphere, such as the N2.

An example of the reaction depicted in scheme 1 below.

Not wishing to be bound to any theory, I believe that the protonation Alliluyeva agent (RCOX) acid, such as sulfuric acid, forms an intermediate compound A. Attaching cationic carbonyl carbon of intermediate compounds And to camptothecin forms an intermediate compound B. then remove molecules HN group B creates the final ester products.

After completion of the reaction, which can be defined by a color change of the solution, the solution may be cooled to room temperature. The solvent may be removed by any commonly known means of separation, such as the evaporation method or the method of filtration. The crude product obtained after removal of solvents, the reaction may be purified using negreanus reflux in an alcohol solvent, such as ethanol. The final product is obtained in crystalline form after recrystallization and/or resultant deposition rates from alcohol.

Got new esters of camptothecin with greatly reduced toxicity, while maintaining antitumor activity, such as crystalline hydrates aliphatic esters, CPT, as a crystalline hydrate camptothecin-20-O-propionate. According to one or more variants of implementation, aliphatic ester CPT can be obtained by using H2SO4-catalyzed acylation reaction shown in the reaction, as shown below:

It is shown that the original camptothecin can interact with propionic anhydride in the catalyst of concentrate sulfuric acid. The reaction mixture can then be redeemed with excess water to remove unreacted propionic anhydride and propionic acid obtained from the reaction. Crystallization of the crude product from absolute ethanol or other solvents, such as other alcohols) gives the final crystalline hydrate camptothecin-20-O-propionate in almost quantitative yield. This would also be true for the other esters CPT of the present invention. It should be understood that the phrase "quantitative output"used in the present description, may VK is ucati outputs 97-100% weight. The purity of the product obtained in the reaction, represents approximately 99-100%. The melting point of the product is approximately 240-243°C.

The present invention will be further explained using the following examples, which are typical for the present invention.

EXAMPLES

Example 1

Crystalline hydrate camptothecin-20-propionate (CZ48) was prepared as follows. Approximately 20 g of camptothecin (0,05747 mol) and approximately 100 ml of propionic anhydride (97%, Aldrich Chemical Co., Milwaukee, WI) was added to a 200-ml round-bottom flask equipped with a magnetic stirrer and a sand bath. The mixture was heated using a sand bath, stirring. Was added dropwise a few drops (8 to 10) of concentrate sulfuric acid (95-98%, reagent A.C.S., Aldrich Chemical Co.), when the temperature of the sand bath reached 80°C. the Mixture then was stirred at 110±10°C overnight (~14 hours). After cooling to room temperature, the reaction mixture parts was poured into 1000 ml of ice water with stirring. After stirring for about 45 minutes, the mixture was filtered. The precipitate obtained from filtration, dried in air for 24 hours. The dried crude product was transferred into a 500-ml round bottom flask, equipped with a mantle.

To this crude product was added 200 ml of absolute ethanol (99.5% pure 200 proof, Aldrich Chemical Co.). The mixture was heated under reflux for 2 hours and then cooled to room temperature. Pure product was obtained as crystals after crystallization from ethanol. It was shown that the purity is 99.8 per cent, using high-performance liquid chromatography (HPLC), and it was determined that the melting point (TPL) is 242°C. Dry nitrogen was usually used as atmospheric reactions all reactions to medicines. All glassware was subjected to heat treatment at 70±10°C for at least 2 h before use. The melting point was observed using the device for determination of melting points MEL - TEMP® and hasn't been fixed. Camptothecin ordered from China and used in this form.

Determination of antitumor activity in vivo. Tumors of the person used in the experiments discussed in the present description, include 1 line of the bladder, 3 lines of breast cancer, 4 lines of the colon, 1 line DSRCT, 1 line kidney, 2 lines of melanoma, line 2 light, 7 lines of the pancreas and 1 line of the prostate. Hydrated crystalline drug CZ48 finely suspended in cottonseed oil and kept in the refrigerator for use. The oral suspension was administered to mice with human tumors and, once a day continuously for 5 days and 2 days or once a day for seven days during the whole period of treatment. The period of treatment for various lines of tumors has changed, ranging from two weeks to one year. Of these proven lines of tumors 18 lines reached a positive response, in other words, more than 50% inhibition of tumor growth with the regime 5 days and 2 days, and 2 line pancreatic tumor had not received a positive response. In continuous mode (daily treatment) one of those two lines tumor of the pancreas when the mode 5/2 with a negative response was achieved positive answer. Table 1 summarizes the results.

Table 1
Antitumor activity of hydrated crystalline CZ48 against human xenografts in Nude mice
TumorColonMammary glandEasyMelanomaPojulu exploration ironBladderDSRCT
ReplyPositive a4/43/32/22/26/71/11/1
Negativeb1/7
a. A positive response indicates the inhibition of growth by more than 50%.
b. A negative response indicates inhibition of growth less than 50%.

While a positive response was defined by the growth inhibition of at least 50%, in fact, achieved complete inhibition of growth for many tumor lines. The effective dose required to achieve a positive response, vary depending on the types of tumors. For CLO-breast carcinoma, the most sensitive lines of the tumor from the test Bank tumors, complete inhibition was achieved with a low dose of hydrated crystalline CZ48, such as 4 mg/kg, whereas to achieve a positive response to CAK-buds, PANC1 pancreatic and SU86.86 pancreatic, 3 of less sensitive lines, required is alas dose is so high, as 1000 mg/kg

Example 2

Determination of antitumor activity in vivo. All animal experiments were carried out on bare Swiss NIH mice, a strain with high fertility. They were fed and kept in the laboratory under strict sterile conditions. To determine the antitumor activity of tumor xenograft grown in naked mice, approximately 1 cm3removed surgically sterile, finely crushed using scissors to iridectomy and suspended in the medium of cell culture in the ratio of 1:10./about. One-half of 1 ml of this suspension, containing approximately 50 mg of crushed tumors in the wet state was subcutaneously injected in the upper part of the dorsal thorax of the mouse. Used groups of four or five animals. The hydrated crystalline CZ48 finely suspended in cottonseed oil and then injected into the cavity of the stomach of a mouse through the anterior abdominal wall using a needle 26 size. Weekly mode, used for oral administration of hydrated crystalline CZ48, was a once a day for 7 days, or five days and two days. This mode was used for all experiments on animals. Treatment was started when tumors reached a volume of approximately 200 mm3, that is well vascularized, paddymac the measurement and grown exponentially. Tumors grown in animals, were examined and measured using a compass once a week. Effective doses were found, when they reached a positive response in mice. The results are shown in table 2.

Table 2
Effective doses of hydrated crystalline CZ48 in lines and tumors
Line tumorsEffective dose (mg/kg)TIand
Bladder-BOL20010
Mammary gland-CLO4500
-MUR3007
-WAR20010
Colon-HT295040
-McC3007
-SQU3067
-SW4810020
DSRCT-MYE10020
Kidney-CAK10002
Easy-DOY15133
-SPA10020
Melanoma-BRO10020
-FOSS3007
Pancreas-ASPC13007
-HS766T10020
-LIE20
-MiaPaCa210020
-PANC110002
-SU86.8610002
The prostate gland-PC3P623007
A. TEE was calculated on the basis of the most acceptable dose of 2000 mg/kg

As shown in table 2, the crystalline hydrate camptothecin-20-O-propionate has a wide therapeutic index in the range from 2 to three resistant lines up to 500 for the most sensitive line (CLO-chest), if you think that 2000 mg/kg is the most acceptable dose.

Example 3

Determination of toxicity in vivo. Groups of 4 or 5 animals of approximately the same age and with similar weight were selected and treated with crystalline hydrate CZ48 oral doses of 1500 mg/kg and 2000 mg/kg, respectively, continuously for 60 days. 3 groups of mice selected for toxicity studies were healthy, the same age and weight and had the tumors. One group was used as control and the other two groups were treated with drugs at two levels of high doses of 1500 mg/kg and 2000 mg/kg, respectively, for approximately two months. Changes of body weight of the animals during treatment were recorded for the group, which is treated, in relation to the group that is not treated, starting from 0 day to the 60th day. The results are shown in table 3.

32,3 (0,8)
Table 3
The toxicity of crystalline hydrate CZ48 in naked miceand
DaysThe body weight (in grams)
ControlbDose 1
(1.5 g/kg)b
Dose 2
(2.0 g/kg)b
0 (Initial date)32,0 (1,4)31,9 (2,0)32,3 (3,1)
1031,9 (1,5)30,4 (3,1)30,7 (4,3)
2132,2 (1,7)31,2 (1,9)31,2 (3,9)
3532,3 (2,1)30,8 (3,2)
4532,1 (3,7)32,8 (1,7)31,7 (2,9)
6032,9 (1,7)33,0 (1,4)30,9 (3,3)
A. The scale used in determining the weight to calibrate and certify annually.
b. Numbers in parentheses represent the standard deviation (SD).

Most strikingly, no significant toxicity in mice, which treat, was not observed in any of the tested dose levels. Loss of body weight was used as a parameter for measuring the toxicity of the agent. Typically the body weight loss of 10% or more in mice during treatment is considered a sign of toxicity. More significant body weight loss shows greater toxicity. Treatment did not lead to a significant loss of body weight, which meant that the applied dose levels were non-toxic to mice and that mice could tolerate doses higher than 2000 mg/kg Continuous treatment of mice with crystalline hydrate camptothecin-20-O-propionate showed a wider range of therapeutic index and did not lead to any significant toxicity.

P the emer 4

Determination of PK parameters of crystalline hydrate CZ48. The pharmacokinetic profile of the crystalline hydrate camptothecin-20-O-propionate with a single dose of 2000 mg/kg was detected after oral administration of naked mice. The corresponding Cmaxand Tmaxwere 284,86±85,55 ng/ml and 2.0±0,0 h, respectively. Camptothecin-20-O-acetate, similar CZ48, which was not a crystalline hydrate was used as an internal standard for the determination of all the PK parameters of hydrate CZ48. 100 μl of mouse plasma were processed using crystalline hydrate CZ48 at a dose of 2000 mg/kg, was transferred to a 2-ml test tube, and then 100 ál of working internal standard solution (400 ng/ml) was also added to the test tube. To the mixture was also added 200 μl of 1%aqueous solution of acetic acid and 1 ml of ethyl ether. After vortex mixing for 10 s, and the mixture was stood at room temperature on a shaker for 10 minutes and then centrifuged at 10,000×g for 15 minutes. The top layer, obtained by centrifugation, was transferred into a clean test tube and evaporated to dryness using an evaporator at 40°C in a stream of nitrogen. The residue was dissolved in 200 µl of solvent system of water/acetonitrile (50/50, vol/about.) and 20-µl aliquot was injected into the HPLC system for analysis. For the study, 100 μl of mouse plasma, which was not subjected to treatment, were treated in the same pic is BOM, how and under-treated. Important PK parameters of hydrate CZ48 and CPT 48 mice were obtained from the analysis of HPLC and shown in table 4.

Table 4
Pharmacokinetic parameters of hydrated crystalline CZ48 and its main metabolite of CPT in mice after oral dose of 2000 mg/kg
OptionsThe hydrated crystalline CZ48CPT
AUC0-8(ng·h·ml-1)---136,43±26,94
AUC0-24(ng·h·ml-1)1927,66±113,92---
AUC0-∞(ng·h·ml-1)2233,44±396,05146,70±34,77
Cmax(ng/ml)284,86±85,5542,28±6,72
Tmax(h)2,0±0,01,0±0,0
T1/2(h)to 8.70±4,181,87±0,63
Ke(1/h)0,9±0,03 0,40±0,13
MRT (h)are 11.62±4,343,36±0,87

Example 5

Determination of the percentage absorption of the crystalline hydrate CZ48 a mouse. Studies of absorption also conducted with crystalline hydrate camptothecin-20-O-propionate using the methods of perfusion of mice 4 hairless mice. Before the methodology for a single perfusion in situ prepared HBSS buffer (pH 7,4), consisting of 9,801 g/l powder HBSS, 0,372 g/l NaHCO3, 3,502 g/l glucose, 5,963 g/l HEPES and 1,164 g/l NaCl, and stored in the refrigerator. The perfusion solution for techniques was obtained by mixing a certain amount of medicinal product, crystal, hydrate, CZ48 with the amount of HBSS buffer. The mixture was heated to 37°C (body temperature) directly before use. A group of 4 mice weighing between 25 to 30 grams and age ranging from 9 to 11 months was anestesiology and was placed on a heating plate. The plate is maintained a constant temperature of 37°C. After pre-perfusion system with a mouse, the two parts of the bowel (small intestine, IS (in situ), and colon) at the same time perfesional freshly prepared perfusion solution. The pump is maintained at a constant flow rate 0,191 ml/min throughout the perfusion. Two samples of the perfusion solution was taken from each of the first output (the small intestine (SI) or the colon) every 30 minutes. The concentration of the drug in the perfusion solution at the outputs were determined using HPLC analysis, and the relevant percentage ratios of the absorbances were calculated relative to the initial amount of drug, and the results are shown in table 5.

Table 5
The results of the perfusion of the gastrointestinal tract in mice
Mouse% absorption of the crystalline hydrate CZ48
SI (small intestine)Colon
No. 116,51-35,17
29,34-73,24
345,856,82
411,498,04
Average22,23-19,46
Standard deviation (SD)16,9638,83

Table 5 shows that significant absorption Ave is proceeded in the small intestine. Negative values of the percentage absorption registered for the colon indicate that the number of drugs coming out of the colon, was more than the amount originally placed in the mouth of the duodenum of the mouse. This can be explained by the accumulation of the drug in the colon as a result of repeated cycles of absorption/outflow.

The crystal structure of the crystalline hydrate CZ48. X-ray analysis of the single crystal was measured using a diffractometer Siemens SMART, has a light square controlled by a computer. A crystal with dimensions of 0.4×of 0.08×0.02 mm was placed in a glass in a stream of cold nitrogen gas at -60°C. Monochromatic radiation of Mo Kα1(λ=0,71073 Å) was used to collect all hemispherical data using the method of the narrow frame. Data were combined using the Siemens SAINT, and the intensities were corrected for Lorentz factor, polarization, absorption, air absorption due to the change in length of the trajectory. Applied empirical amendments and averaged excessive deflection. Final cell parameters were refined using deviations 1971 with I>10 σ(I). The parameters of the tetragonal cell are a=15,008(2) Å, b=6,977(1) Å, c=21,810(3) Å, β=99,959°, V=2249,2(5) Å3, Z=4, ρ=1,354 fuel 3, 2θmax=56,66°. The structure was solved by direct methods with the space group P21(No. 4) and refined using full-fabric calculations by the method of least squares F2thermal movement of all C, N, O atoms were considered anisotropically. Final R indices [I > 2σ(I)], R1=0,0454, wR2=0,0763, R indexes [all data], R1=0,1105, wR2=0,0933. All calculations were performed using the software package Siemens SHELXTL. Data about crystal and structure refinement for [(C23H20N2O5]·3H2O are shown in table 7. Atomic coordinates (×104) and equivalent isotropic parameters displacement (Å2×103for [C23H20N2O5]·3H2O are shown in table 8. The bond length [Å] and angles [°] for [(C23H20N2O5]·3H2O are shown in table 9. The parameters of the anisotropic displacement (Å2×103for [C23H20N2O5]·3H2O are shown in table 10.

Table 7
Data about crystal and structure refinement for [(C23H20N2O5]·3H2O
Identification codeT94
Empirical formulaC23H26N2 O8
Molecular weight by the formula458,46
Temperature213(2) K
Wavelength0,71073 Å
Crystal systemMonoclinically
Space groupP21
Permanent grillsa=15,008(2) Åα=90°
b=6,9769(10) Åβ=99,959(2)°
c=21,810(3) Åγ=90°
Volume2249,2(5) Å3
Z4
Density (calculated)1,354 g/cm3
To the ratio of absorption 0,103 mm-1
F(000)968
The crystal size0,40×0,08×0.02 mm3
Theta range for data collectionFrom 1.53 to 28.33°
The ranges of the index-20<=h<=15, -8<=k<=9, -28<=l<=21
Collected reflections14162
Independent reflections14162 [R(int)=0,0496]
Completeness to theta=28,33°95.6%of
Amendment absorptionEmpirical
Max. and min. transmission0,8092 and 0,7004
Method of refiningFull-fabric method of least squares F2
Data/restraints/ parameters14162/1/633
The criterion of consent F20,78
Final R indices [I > 2sigma(I)]R1=0,0454, wR2=0,0763
R indices (all data)R1=0,1105, wR2=0,0933
The extinction coefficient0,00000(11)
The highest diffraction peak and failurehas 0.168 and -0,172 e.Å-3

td align="center"> 695(2)td align="center"> 1050(1)
Table 8
Atomic coordinates (×104) and equivalent isotropic parameters displacement (Å2×103for [C23H20N2O5]·3H2O
xyzU(EQ)
C(1)-1249(2)734(5)1218(2) 45(1)
C(2)-2180(2)697(5)1137(2)51(1)
C(3)-2705(2)747(5)543(2)54(1)
C(4)-2300(2)845(5)31(2)47(1)
C(5)-1338(2)864(5)89(2)37(1)
C(6)-816(2)826(4)697(2)38(1)
C(7)-894(2)886(4)-432(2)39(1)
C(8)27(2)877(4)-327(2)33(1)
C(9)491(2)903(4)291(1)30(1)
C(10)831(5)-764(1)36(1)
C(11)1471(2)938(4)284(2)33(1)
C(12)2388(2)759(5)-528(2)35(1)
C(13)3157(2)772(4)-34(2)34(1)
C(14)3067(2)920(4)572(1)32(1)
C(15)2199(2)993(4)744(1)35(1)
C(16)4063(2)652(5)- 223 is(1)39(1)
C(17)4698(2)-38(5)863(2)43(1)
C(18)3920(2)1101(5)36(1)
C(19)4191(2)3246(4)1112(1)39(1)
C(20)5039(2)3639(5)1599(2)53(1)
C(21)3547(2)-1319(5)1731(2)42(1)
C(22)3391(2)-1700(5)2375(2)55(1)
C(23)3057(2)-3713(5)2462(2)69(1)
C(24)-1296(2)7496(5)6345(2)43(1)
C(25)-2208(2)7380(5)6318(2)50(1)
C(26)-2779(2)7338(5)5734(2) 52(1)
C(27)-2434(2)7452(5)5201(2)46(1)
C(28)-1494(2)7557(4)5214(2)33(1)
C(29)-919(2)7572(4)5799(2)34(1)
C(30)-1101(2)7601(4)4672(2)37(1)
C(31)-179(2)7612(5)4735(2)31(1)
C(32)338(2)7628(5)5341(2)30(1)
C(33)448(2)7597(5)4262(1)36(1)
C(34)1296(2)7576(5)5291(2)32(1)
C(35)2136(2)7419(4)4429(2)35(1)
C(36)2932(2)7290(4)4903(2)33(1)
C(37)2901(2)7376(4)5517(2)31(1)
C(38)2060(2)7521(4)5730(1)33(1)
C(39)3818(2)7072(5)4672(2)45(1)
C(40)4509(2)6193(5)5742(2)42(1)
C(41)3786(2)7434(5)5964(1)32(1)
C(42)4111(2)9549(4)6027(2)42(1)
C(43)485(2) 9880(5)6493(2)52(1)
C(44)3461(2)4998(5)6663(2)37(1)
C(45)3447(2)4602(5)7329(2)51(1)
C(46)3242(2)2520(5)7463(2)63(1)
N(1)120(2)881(4)797(1)38(1)
N(2)1565(2)841(4)-331(1)33(1)
N(3)17(2)7618(4)5863(1)34(1)
N(4)1344(2)7534(4)4668(1)31(1)
O(1)2435(1)639(3) -1088(1)44(1)
O(2)4756(1)-117(3)255(1)49(1)
O(3)5289(2)-760(3)1231(1)57(1)
O(4)3788(1)526(3)1658(1)39(1)
O(5)3469(1)-2474(3)1308(1)54(1)
O(6)2142(1)7384(3)3869(1)45(1)
O(7)4502(1)6092(3)5127(1)48(1)
O(8)5108(1)5412(3)6091(1)55(1)
O(9)3704(1)6866(3)6584(1)35(1)
O(10)3291(1)3905(3)6238(1)47(1)
O(11)998(2)8164(4)2777(1)59(1)
O(12)1226(2)1248(4)2025(1)56(1)
O(13)602(2)4974(4)2037(1)68(1)
O(14)1264(2)7001(4)7044(2)63(1)
O(15)802(2)3200(5)7072(2)83(1)
O(16)-1229(3)5058(4)2183(2)59(1)
U(EQ) is defined as one third of the trace orthogonalizing tensor Uij

Table 9
The bond length [Å] and angles [°] for [(C23H20N2O5]·3H2O
C(1)-C(2)1,378(4)
C(1)-C(6)1,404(4)
C(2)-C(3)1,393(5)
C(3)-C(4)1,362(4)
C(4)-C(5)1,428(4)
C(5)-C(7)1,413(4)
C(5)-C(6)1,418(4)
C(6)-N(1)1,385(3)
C(7)-C(8)1,362(4)
C(8)-C(9)1,407(4)
C(8)-C(10)1,498(4)
C(9)-N(1)1,317(4)
C(9)-C(11)1,473(4)
C(10)-N(2)1,473(3)
C(11)-C(15)1,351(4)
C(11)-N(2)1,375(4)
C(12)-O(1)1,238(3)
C(12)-N(2)1,377(3)
C(12)-C(13)1,437(4)
C(13)-C(14)1,355(4)
C(13)-C(16)1,490(4)
C(14)-C(15)1,418(4)
C(14)-C(18)1,510(4)
C(16)-O(2)1,443(3)
C(17)-O(3)1,199(3)
C(17)-O(2)1,345(4)
C(17)-C(18)1,526(4)
C(18)-O(4)1,433(3)
C(18)-C(19)1,551(4)
C(19)-C(20)1,536(4)
C(21)-O(5)1,215(4)
C(21)-O(4)1,354(4)
C(21)-C(22)1,487(4)
C(22)-C(23)1,515(5)
C(24)C(25)1,361(4)
C(24)-C(29) 1,405(4)
C(25)C(26)1,409(5)
C(26)C(27)1,354(4)
C(27)C(28)1,409(4)
C(28)-C(30)1,408(4)
C(28)-C(29)1,413(4)
C(29)-N(3)1,388(3)
C(30)-C(31)1,368(4)
C(31)C(32)1,411(4)
C(31)-C(33)1,512(4)
C(32)-N(3)1,311(4)
C(32)-C(34)1,460(4)
C(33)-N(4)1,477(3)
C(34)-C(38)1,362(4)
C(34)-N(4)1,373(4)
C(35)-O(6)1,223(3)
C(35)-N(4)1,380(3)
C(35)C(36)1,440(4)
C(36)C(37)1,348(4)
C(36)-C(39)1,508(4)
C(37)C(38)1,423(4)
C(37)-C(41)1,507(4)
C(39)-O(7)1,468(3)
C(40)-O(8)1,204(3)
C(40)-O(7)1,340(4)
C(40)C(41)1,531(4)
C(41)-O(9)1,434(3)
C(41)C(42)1,553(4)
C(42)C(43)1,532(4)
C(44)-O(10)1,192(4)
C(44)-O(9)1,372(4)
C(44)C(45)1,483(4)
C(45)C(46)1,524(4)
C(2)-C(1)-C(6)to 119.8(3)
C(1)-C(2)-C(3)121, 1million(3)
C(4)-C(3)-C(2)120,1(3)
C(3)-C(4)-C(5) to 121.0(3)
C(7)-C(5)-C(6)119,5(3)
C(7)-C(5)-C(4)122,6(3)
C(6)-C(5)-C(4)to 117.9(3)
N(1)-C(6)-C(1)118,3(3)
N(1)-C(6)-C(5)the level of 121.8(3)
C(1)-C(6)-C(5)119,9(3)
C(8)-C(7)-C(5)118,1(3)
C(7)-C(8)-C(9)118,7(3)
C(7)-C(8)-C(10)131,8(3)
C(9)-C(8)-C(10)to 109.6(3)
N(1)-C(9)-C(8)126,2(3)
N(1)-C(9)-C(11)125,2(3)
C(8)-C(9)-C(11)108,6(3)
N(2)-C(10)-C(8)102,1(2)
C(15)-C(11)-N(2)121,3(3)
C(15)-C(11)-C(9)132,3(3)
N(2)-C(11)-C(9)106,4(3)
O(1)-C(12)-N(2) 121,2(3)
O(1)-C(12)-C(13)to 124.4(3)
N(2)-C(12)-C(13)114,3(3)
C(14)-C(13)-C(12)122,0(3)
C(14)-C(13)-C(16)to 121.6(3)
C(12)-C(13)-C(16)116,4(3)
C(13)-C(14)-C(15)120,8(3)
C(13)-C(14)-C(18)117,7(3)
C(15)-C(14)-C(18)121,5(3)
C(11)-C(15)-C(14)117,7(3)
O(2)-C(16)-C(13)113,5(3)
O(3)-C(17)-O(2)118,5(3)
O(3)-C(17)-C(18)to 123.5(3)
O(2)-C(17)-C(18)to 117.9(3)
O(4)-C(18)-C(14)112,3(2)
O(4)-C(18)-C(17)109,7(3)
C(14)-C(18)-C(17)111,6(3)
O(4)-C(18)-C(19)105,4(3)
C(14)-C(18)C(19) 108,6(3)
C(17)-C(18)-C(19)108,9(3)
C(20)-C(19)-C(18)114,0(3)
O(5)-C(21)-O(4)122,3(3)
O(5)-C(21)-C(22)126,1(3)
O(4)-C(21)-C(22)111,6(3)
C(21)-C(22)-C(23)of 113.2(3)
C(25)-C(24)-C(29)to 121.0(3)
C(24)C(25)C(26)119,4(4)
C(27)-C(26)-C(25)120,9(3)
C(26)C(27)C(28)121, 1million(3)
C(30)-C(28)-C(27)of 123.2(3)
C(30)-C(28)-C(29)118,6(3)
C(27)C(28)-C(29)118,1(3)
N(3)-C(29)-C(24)117,8(3)
N(3)-C(29)-C(28)122,6(3)
C(24)-C(29)-C(28)119,5(3)
C(31)-C(30)-C(28)118,6(3)
C(30)-C(31)C(32)118,5(3)
C(30)-C(31)-C(33)132,1(3)
C(32)-C(31)-C(33)109,4(3)
N(3)-C(32)-C(31)126,0(3)
N(3)-C(32)-C(34)125,3(3)
C(31)C(32)-C(34)108,6(3)
N(4)-C(33)-C(31)101,6(2)
C(38)-C(34)-N(4)120,8(3)
C(38)-C(34)-C(32)132,0(3)
N(4)-C(34)-C(32)107,1(3)
O(6)-C(35)-N(4)122,2(3)
O(6)-C(35)C(36)124,5(3)
N(4)-C(35)C(36)of 113.2(3)
C(37)-C(36)-C(35)122,9(3)
C(37)-C(36)-C(39)121,3(3)
C(35)C(36)-C(39)115, 8mm(3)
C(36)C(37)C(38)120,9(3)
C(36)C(37)-C(41)117,8(3)
C(38)-C(37)-C(41)121,2(3)
C(34)-C(38)-C(37)117,3(3)
O(7)-C(39)-C(36)112,1(3)
O(8)-C(40)-O(7)119,0(3)
O(8)-C(40)C(41)of 123.2(3)
O(7)-C(40)C(41)117,6(3)
O(9)-C(41)-C(37)of 113.2(2)
O(9)-C(41)-C(40)108,9(3)
C(37)-C(41)-C(40)111,9(3)
O(9)-C(41)C(42)104,9(2)
C(37)-C(41)C(42)108,1(3)
C(40)C(41)C(42)109,7(2)
C(43)-C(42)-C(41)USD 114.9(3)
O(10)-C(44)-O(9)122,5(3)
O(10)-C(44)C(45)127,1(3)
O(9)-C(44)C(45)110,4(3)
C(44)C(45)C(46)113,8(3)
C(9)-N(1)-C(6) by 115.7(3)
C(11)-N(2)-C(12)123,8(3)
C(11)-N(2)-C(10)113,3(2)
C(12)-N(2)-C(10)122,9(3)
C(32)-N(3)-C(29)115,6(3)
C(34)-N(4)-C(35)of 124.8(3)
C(34)-N(4)-C(33)of 113.2(2)
C(35)-N(4)-C(33)122,0(3)
C(17)-O(2)-C(16)122,4(2)
C(21)-O(4)-C(18)117,1(3)
C(40)-O(7)-C(39)122,4(2)
C(44)-O(9)-C(41)116,6(2)
Conversion of symmetry usually describes identical atoms.

Table 10
The parameters of the anisotropic displacement (Å2×103for [C23H20N2O5]·3H2O
U11U22U33 U23U13U12
C(1)45(2)43(2)48(2)-1(2)9(2)1(2)
C(2)51(2)51(3)55(3)-2(2)18(2)-4(2)
C(3)44(2)45(2)73(3)-7(2)12(2)-4(2)
C(4)41(2)38(2)59(3)-3(2)-4(2)-1(2)
C(5)38(2)24(2)46(2)-4(2)2(2)-7(2)
C(6)40(2)25(2)48(2)(2) 9(2)2(2)
C(7)45(2)28(2)40(2)0(2)-7(2)-6(2)
C(8)39(2)24(2)34(2)-2(2)2(2)-5(2)
C(9)39(2)19(2)31(2)1(2)6(2)1(2)
C(10)44(2)28(2)32(2)1(2)-3(2)-5(2)
C(11)39(2)29(2)31(2)3(2)4(2)1(2)
C(12)43(2)35(2)27(2)1(2) 7(2)-7(2)
C(13)37(2)28(2)36(2)2(2)5(2)2(2)
C(14)38(2)27(2)28(2)-4(2)2(2)1(2)
C(15)44(2)34(2)24(2)3(2)2(2)6(2)
C(16)42(2)45(2)28(2)-1(2)4(2)-3(2)
C(17)40(2)41(2)44(2)-6(2)-1(2)2(2)
C(18)35(2)43(2)28(2)2(2)1(2) 4(2)
C(19)40(2)38(2)35(2)-4(2)-1(2)0(2)
C(20)54(2)53(3)47(2)-4(2)-6(2)-6(2)
C(21)39(2)44(3)40(2)4(2)-1(2)4(2)
C(22)64(3)60(3)40(2)14(2)11(2)9(2)
C(23)63(3)72(3)75(3)19(3)20(2)2(2)
C(24)47(2)38(2)45(2)1(2)10(2)3(2)
C(25)43(2)50(2)61(3)1(2)18(2)0(2)
C(26)39(2)43(2)73(3)0(2)5(2)1(2)
C(27)41(2)34(2)58(3)0(2)-2(2)1(2)
C(28)35(2)21(2)42(2)-2(2)-1(2)1(2)
C(29)36(2)24(2)42(2)3(2)4(2)1(2)
C(30)48(2)22(2)33(2)2(2)-11(2)2(2)
C(31) 34(2)23(2)34(2)-1(2)-2(2)1(2)
C(32)32(2)24(2)33(2)3(2)1(2)0(2)
C(33)46(2)28(2)33(2)2(2)0(2)-3(2)
C(34)42(2)23(2)30(2)-2(2)2(2)1(2)
C(35)50(2)22(2)32(2)0(2)7(2)1(2)
C(36)41(2)27(2)31(2)2(2)8(2)1(2)
C(37)33(2) 23(2)35(2)0(2)2(2)0(2)
C(38)39(2)30(2)28(2)-2(2)4(2)1(2)
C(39)46(2)48(2)41(2)6(2)6(2)0(2)
C(40)43(2)42(2)43(2)2(2)12(2)2(2)
C(41)35(2)36(2)26(2)3(2)7(2)5(2)
C(42)42(2)37(2)44(2)0(2)1(2)-5(2)
C(43)51(2)54(3) 49(2)-6(2)4(2)-9(2)
C(44)37(2)41(2)31(2)4(2)3(2)10(2)
C(45)61(2)50(2)44(2)8(2)13(2)2(2)
C(46)70(3)64(3)55(3)17(2)9(2)-7(2)
N(1)39(2)31(2)43(2)1(2)6(1)-4(1)
N(2)35(2)32(2)30(2)-1(1)1(1)-4(1)
N(3)38(2)26(2)36(2) 0(1)4(1)2(1)
N(4)36(2)28(2)28(2)2(1)1(1)0(1)
O(1)47(1)52(2)33(1)1(1)2(1)-4(1)
O(2)43(2)67(2)37(2)0(1)5(1)15(1)
O(3)49(2)68(2)50(2)0(1)-3(1)23(1)
O(4)44(1)41(2)32(1)1(1)4(1)2(1)
O(5)78(2)40(2)40(2)0(1) 3(1)2(1)
O(6)57(2)52(2)26(1)-1(1)8(1)3(1)
O(7)45(1)63(2)38(2)3(1)14(1)13(1)
O(8)47(2)68(2)49(2)5(1)7(1)22(1)
O(9)39(1)33(1)32(1)-2(1)4(1)1(1)
O(10)62(2)43(2)34(2)-5(1)2(1)3(1)
O(11)63(2)58(2)48(2)-2(2)-10(2) -1(2)
O(12)62(2)64(2)38(2)2(1)1(1)-1(1)
O(13)58(2)69(2)73(2)-6(2)1(2)9(2)
O(14)63(2)70(3)52(2)12(2)0(1)2(2)
O(15)59(2)89(3)94(3)24(2)-5(2)1(2)
O(16)67(2)59(3)46(2)-2(2)0(1)2(2)
The exponent of the factor anisotropic displacement is of the form:
-2π2[h2a*2U11+...+2hka*b*U12]

From these studies it is evident that the compounds nastojasih the invention show a remarkable level of anticancer activity. It can be applied to the spectrum of the analyzed tumors, as well as to the quality of the answers. The method of the present invention is able to completely block the growth and completely regress the xenografts of human carcinoma (e.g., lung, breast, colon, stomach, pancreas, bladder, prostate, sarcoma and ovarian cancer) and malignant melanomas. This was achieved without any observed toxicity. Many of the mammals, which was continuously treated for six months, showed no negative effects, no resumption of tumor growth, which they had once been. Hydrated crystalline aliphatic esters of the present invention should all have the same degree of effectiveness and therapeutic value of the index.

Applicants definitely include the full contents of all cited references in the disclosure of this application. Additionally, when an amount, concentration, or other value or parameter is shown as either a range, preferred range, or a list of upper preferable values and lower preferable values, should be understood as a particular way disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower is thinned range or preferred value, no matter disclosed whether the ranges separately. If the range of numeric values listed in the present description, unless otherwise stated, assume that the range includes the endpoints, and all integers and fractions within the range. I believe that the possibilities of the invention are not limited to the values listed in the definition range.

Other embodiments of the present invention will be obvious to experts in the art from consideration of this specification and practice of the present invention disclosed in the present description. I believe that the present specification and examples be considered as exemplary only with the true scope and essence of the invention defined in accordance with the following claims and its equivalents.

1. Crystalline hydrate camptothecin formula (I):

in which n includes a number ranging from 1 to 10, and R1represents a C2-C6alkyl group.

2. Crystalline hydrate camptothecin according to claim 1, in which R1is a linear C2-C4alkyl group.

3. Crystalline hydrate camptothecin according to claim 1, in which R1represents ethyl.

4. Crystalline hydrate camptothecin according to claim 3, in which crystallizes the th hydrate camptothecin has a melting point of from about 240°to about 243°C.

5. Crystalline hydrate camptothecin according to claim 3, in which the compound has a melting point of about 242°C.

6. Crystalline hydrate camptothecin of claim 1, wherein the compound has a purity of from about 99% to about 100%.

7. Crystalline hydrate camptothecin of claim 1, wherein n comprises a number ranging from 1 to 3.

8. A method of treating cancer or a malignant tumor in a patient, comprising introducing a composition containing an effective amount of the crystalline hydrate camptothecin according to claim 1, wherein said cancer or a malignant tumor sensitive to the specified composition.

9. The method according to claim 8, in which R1represents ethyl.

10. The method of claim 8, wherein said cancer is a leukemia, melanoma, liver cancer, breast cancer, ovarian cancer, prostate cancer, stomach cancer, bladder cancer, desmoplastic small-round-cell tumor (DSRCT), pancreatic cancer, lung cancer, kidney cancer, colon cancer or cancer of the Central nervous system.

11. The method according to claim 10, in which R1represents ethyl.

12. The method according to claim 8, in which the crystalline hydrate camptothecin administered intravenously, intramuscularly, or transdermal.

13. The method according to claim 8, in which the crystalline hydrate camptothecin administered orally.

14. The pharmaceutical composition is for the treatment of cancer or malignant tumor, comprising an effective amount of the crystalline hydrate camptothecin according to claim 1 and a pharmaceutically acceptable carrier.

15. The pharmaceutical composition according to 14, in which the carrier is a lipid or lepidoptery emulsifier.

16. Crystalline hydrate camptothecin according to claim 1, wherein said crystalline hydrate camptothecin has a therapeutic index of from 2 to 500, as defined in the introduction of this crystalline hydrate camptothecin naked mice with tumor xenograft, using a dose of 2000 mg/kg as the highest permissible dose.

17. Crystalline hydrate camptothecin according to item 16, wherein said therapeutic index represents from 10 to 100.

18. Crystalline hydrate camptothecin according to claim 1, in which the crystalline hydrate camptothecin has monoclinically system sizes (from 0.10 to 0.50)×(0.01 to 0.10)×(0.01 to 0.05) mm3and volumes from 100 to 5000 Å3.

19. Crystalline hydrate camptothecin on p, wherein said crystalline hydrate camptothecin is a crystalline hydrate camptothecin-20-propionate.



 

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9 cl, 12 tbl, 31 ex, 3 dwg

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6 cl, 2 tbl, 22 ex

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4 ex

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13 cl, 4 dwg, 2 tbl, 9 ex

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12 cl, 7 tbl, 14 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to novel azoindolizines or pharmaceutically acceptable salts thereof having Mek-kinase inhibitory activity in formula : ZA means CRA; RA means H or halogen; each of R1, R2 and R3 means H; W means: , each R4 and R5 means H; X1 represents -OR7; each R7 means C2-C12-hydroxyalkyl, 2,3-dihydroxypropyl, C2-C3-alkenoxyC1-C6-alkoxy, (2,2-dimethyl-[1,3]dioxalan-4-yl)-methyl or piperidinyl; X4 means: R6 means halogen or -SR16; R6 means halogen; p is equal to 1; R16 means C1-C12-alkyl.

EFFECT: invention relates to a pharmaceutical compositions containing these compounds, to a method of inhibiting the abnormal cell growth and the use of the compounds for preparing a drug preparation for inhibiting the abnormal cell growth.

9 cl, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a new monophosphate salt of 4-methyl-N-[3-(4-methylimidazol-1-yl)-5-trifluoromethylphenyl]-3-(4-pyridin-3-yl-pyrimidin-2-ylamino)benzamide which may be used for treating a disease responding to protein kinase activity inhibition. The method for preparing the monophosphate salt of 4-methyl-N-[3-(4-methylimidazol-1-yl)-5-trifluoromethylphenyl]-3-(4-pyridin-3-yl-pyrimidin-2-ylamino)benzamide involves the reaction of 4-methyl-N-[3-(4-methylimidazol-1-yl)-5-trifluoromethylphenyl]-3-(4-pyridin-3-yl-pyrimidin-2-ylamino)benzamide in the form of a free base with phosphoric acid in methanol. Also, the invention refers to a pharmaceutical composition containing: (a) a therapeutically effective amount of the monophosphate salt of 4-methyl-N-[3-(4-methylimidazol-1-yl)-5-trifluoromethylphenyl]-3-(4-pyridin-3-yl-pyrimidin-2-ylamino)benzamide; and (b) at least one pharmaceutically acceptable carrier, a diluent, an excipient, as well as to the use of the therapeutically effective amount of the monophosphate salt of 4-methyl-N-[3-(4-methylimidazol-1-yl)-5-trifluoromethylphenyl]-3-(4-pyridin-3-yl-pyrimidin-2-ylamino)benzamide in preparing a drug preparation for treating a disease responding to protein kinase activity inhibition.

EFFECT: preparing the composition for treating a disease responding to protein kinase activity inhibition.

4 cl, 8 dwg, 17 tbl, 11 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a Raf-kinase inhibitor that is a solid form A of 1-methyl-5-(2-(5-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-4-yloxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[(1]imidazole-2-amine hydrate, a pharmaceutical composition containing a solid form A, and the use thereof for treating cancer.

EFFECT: preparing the composition for treating cancer.

21 cl, 48 dwg, 16 tbl, 19 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to medicine, namely oncology and may be used for treating multiple myeloma in a patient recurrent after bortezomib or refractory therapy. That is ensured by administering a VEGF antagonist representing a VEGF antibody (bevacizumab). What is also presented is a method of treating multiple myeloma recurrent after alkylating agent therapy.

EFFECT: group of inventions provides the effectiveness of the VEGF antibody for suppression of bortezomin sensitive and/or resistant tumours.

48 cl, 10 dwg, 2 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science, and aims at treating viral diseases in animals. What is declared is a composition for treating viral diseases in animals, containing two subtypes of vertebrate recombinant interferon mixed in equal molar proportions.

EFFECT: use of the declared composition enables ensuring the substantial increase of the therapeutic effect of recombinant interferons and minimising the potential adverse effects associated with their use.

4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely to an antiproliferative agent for treating benign hyperplasia of prostate (BHP). The antiproliferative agent for treating benign hyperplasia of prostate based on herbal raw materials represents terrestrial peat lipids prepared by grinding terrestrial peat in a planetary ball mill or a ball mill, extracting ground peat in mixed ethanol and chloroform, filtering, evaporating and drying in the specific environment.

EFFECT: agent enables extending the range of herbal products on the basis of domestic herbal raw materials effective in treating BHP.

3 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to an anticancer drug containing a combination of 1-(3-C-ethinyl-β-D-ribopentifuranosyl)cytosine or its salt, and carboplatin. The invention also refers to the use of this combination for preparing the anticancer agent, a method of treating cancer which comprises administering the combination to the patient, and a method of enhancing the anticancer effect of carboplatin.

EFFECT: combination of 1-(3-C-ethinyl-β-D-ribopentifuranosyl)cytosine or its salt, and carboplatin provides the enhanced anticancer effect and the reduced unfavourable effects.

8 cl, 3 dwg, 1 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to immunology. There are presented versions of a CD20 modified antibody or its antigen-binding fragment. Each version is characterised by the fact that it comprises a variable region of a light chain and a variable region of a heavy chain and induces a higher level of apoptosis as compared with B-Lyl murine antibody. There are presented versions of the compositions to enhance the effector functions. One of such compositions contains antibodies wherein at least 20% of the Fc oligosaccharides are bisectional and non-fucosylated, whereas the other one contains antibodies wherein at least 50% of the oligosaccharides are non-fucosylated. Also, there are described: versions of a host cell to produce the antibodies, an expression vector, as well as the versions of the coding polynucleotides, a method for producing the antibodies in a cell and using the antibodies for preparing a drug for treating disorders treated by B-cells depletion.

EFFECT: use of the inventions provides the antibodies with the improved therapeutic properties, including with higher Fc receptor binding and enhanced effector function that can be find application in treating tumors.

36 cl, 3 ex, 9 tbl, 26 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to virology, and concerns the Newcastle disease virus strain. The characterised strain NDV/Mallard/Adigeya/8/2008 is recovered from a dead duck, and deposited in the Collection of Cultures of the State Research Centre for Virology and Biotechnology "Vector" under registration number No. V-512.

EFFECT: presented strain may be used to study the oncolytic properties and mechanisms, and to develop the anticancer candidate preparations based thereon.

9 dwg, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to combinations of peptides in each case with the same sequence length (SEQL) which can be prepared in a stable reproducible quality and quantity of a mixture (A) containing a number of x amino acid with protected acid groups or a number of z peptides with the acid groups protected by the protective groups and the activated amino groups, with the amino acids in the mixture (A) found in a specific molar ratio, and a mixture (B), containing a number of y amino acids with the amino groups protected by the protective groups, with a molar ratio of the amino acids of the mixture (B) being the same as the molar ratio of the amino acids of the mixture (A), and the number x=y, and x is a figure from 11 to 18.

EFFECT: new combinations of the peptides are presented.

13 cl, 2 dwg, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine, in particular to pharmacology, to medication, possessing wound-healing activity. Application of hipaconitine as medication, possessing wound-healing activity is disclosed.

EFFECT: hipactonitin possesses expressed wound-healing activity.

6 tbl, 1 ex

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