Polymer systems of delivering active substances
SUBSTANCE: invention relates to compositions, containing an active substance. Described is a composition for the active substance delivery, which contains: a) at least, one block-copolymer, containing, at least, one poly(2-oxazoline) block A, consisting of repeating units of formula where RA stands for a hydrocarbon group, which can be optionally substituted with -OH, -SH, -COOH, -NR'2, -COOR', -CONR', -CHO, where R' stands for H or C1-3 alkyl, and RA is selected in such a way that a repeating link of formula (I) is hydrophilic; and at least one poly(2-oxazoline) block B, consisting of repeating units of formula (II), where RB stands for a hydrocarbon group, which can be optionally substituted with halogen, -OH, -SH, -COOH, -NR''2, -COOR'', -CONR'', -CHO, where R'' stands for H, alkyl or alkenyl, and RB is selected in such a way that a repeating link of formula is more hydrophobic than the repeating link of formula (I); and (b) one or more active substances. Also described is application of a copolymer for solubilisation of the active substance in water or a water solution.
EFFECT: described copolymers serve as a universal medication delivery system with high loading.
12 cl, 6 dwg
This invention relates to compositions and preparations containing the active substance, in particular a hydrophobic active substances, together with the delivery system, which contributes to the dissolution of these substances.
Many of the most effective medicinal substances and potential medicinal substances insoluble in water. The drug is poorly soluble drugs such as paclitaxel (PTX) with a solubility of approximately 1 μg/ml, remains a major problem in drug delivery (Huh, K. M., et al., J. Controlled Release 126, 122-129 (2008); Dabholkar, R. D., et al. Int. J. Pharm. 315, 148-157 (2006); Yang, T., et al., Int. J. Pharm. 338, 317-326 (2007); Torchilin, V.P., Cell. Mol. Life. Sci 61, 2549-2559 (2004); Haag, R., Angew. Chem. Int. Ed. 43, 278-282 (2004)). Currently, clinical drug paclitaxel, Taxol®, contains less than 1% wt./wt. current medicinal substances and 99% wt./wt. media that causes significant side effects in patients. Such problems have active substances, used in other technical fields, such as plant protection against parasites, etc. we developed several methods of solubilization or dispersion of the active substances. Traditional methods are usually based on the use of solvents, surfactants or chelat forming substances. These methods have one or more shortcomings associated with Tox is the durability of the media, limited stability of drugs in the aquatic environment, in particular, when diluted, or complexity of the formulation process.
Recently, liposomes (Wu, J., et al., Int. J. Pharm. 316, 148-153 (2006)), micro - and nanoparticles (Desai, N.P. et al., Anti-Cancer Drugs 19, 899-909 (2008)), and polymeric micelles (Huh, K. M., et al., J. Controlled Release 126, 122-129 (2008); Konno, T., et al., J. Biomed. Mat. Res., Part A, 65A, 210-215 (2002); Kim, S.C., et al., J. Controlled Release 72, 191-202 (2001)) has been studied intensively as solubilization/drug delivery. Each of these approaches has its advantages and disadvantages. One of the main limitations of polymeric micelles is the capacity and the total number of medicinal substance that can be dissolved. The patent application U.S. 20040185101 discloses polymeric compositions with the ability to solubilisate hydrophobic drug substances in the aquatic environment. However, the capacity of these compositions is limited for example to <10% wt./wt. for paclitaxel, or less than 1% wt./wt. for cyclosporine A.
Poly(2-oxazoline)s recently attracted special attention for applications in Biomedicine. In particular, great interest hydrophilic poly(2-methyl-oxazoline)s p(Meox) and poly(2-ethyl-2-oxazoline)s p(Etox), as they have a mask (Zalipsky, S., et al., J. Pharm. Sci. 85, 133-137 (1996); Woodle, M. C., et al., Bioconjugate Chem. 5, 494-496 (1994)) and protein-repellent properties of the mi (Konradi, R., et al., Langmuir 24, 613-616 (2008)) and subjected to rapid renal clearance (Gaertner, F.C., et al., J. Controlled Release 119, 219-300 (2007)), similar to properties of poly(ethylene glycol), a widely used polymer in injectable delivery systems of drugs.
The present invention is to provide compositions containing the delivery system, which allows you to effectively solubilisate and/or to formulate active ingredients, in particular hydrophobic active substances. In particular, the compositions should be easy to manufacture and have high loading capacity for a specific active substance.
In order to solve the problem, the present invention provides compositions containing
(a) at least one copolymer containing the repeating unit of the formula (I),
where RAmeans hydrocarbon group, optionally substituted by-OH, -SH, -COOH, -NR'2, -COOR', -CONR', -CHO, where R' represents H or C1-3alkyl, and RAis chosen so that the repeating element of the formula (I) is hydrophilic,
and the repeating unit of the formula (II)
where RBmeans hydrocarbon group, optionally substituted with halogen, -OH, -SH, -COOH, -NR2, -COOR", -CONR", -CHO, where R ' denotes H, alkyl, alkenyl, and RBset the m way that recurring element of the formula (II) are more hydrophobic than the repeating element of the formula (I); and
(b) one or more active substances.
Moreover, this invention provides the above copolymers. Compositions according to the invention can be used in various technical fields, including pharmaceuticals, diagnostics (including animal health and plant protection.
Figure 1 is apresents fluorescence intensity and attitude of I1/I3solutions purine depending on the concentration of specific copolymers used in the context of the invention, at 25°C.
Figure 2 presents the results of the dissolution of PTX with amphiphilic copolymers method of thin films.
Figure 3 presents the results of measurements of dynamic light scattering filled medicine micelles formed by the compositions according to this invention.
4 shows the measurement results of the cytotoxicity of the polymers used in this invention, in various cell lines.
Figure 5 presents the dependence of the viability of the cells with multidrug resistance line MCF7/ADR, depending on the dose of paclitaxel, in mixtures prepared according to the invention.
Figure 6 presents the relative weight of tumors (a) and suppression of tumor growth (C) Rel is relatively negative control mice, the treated compositions prepared according to the invention and a commercial product.
As mentioned above, in the units of the formula (I) RAis a hydrocarbon group, optionally substituted by-OH, -SH, -COOH, -NR'2, -COOR', -CONR', -CHO, where R' represents H or C1-3alkyl, and RAchosen so that the repeating element of the formula (I) is hydrophilic.
Preferably, as RAto choose C1-8hydrocarbon group, preferably, C1-6hydrocarbon group, more preferably, C1-3hydrocarbon group, and in particular, C1-2hydrocarbon group, all of which may be optionally substituted. Preferred hydrocarbon groups are alkyl groups.
It is clear that the hydrophilic property of the link of the formula (I)as described above, will depend on the size of the hydrocarbon groups of RA. When choosing a short hydrocarbon group such as methyl or ethyl, the group RA, unsubstituted or substituted by the above substituents, there will always be hydrophilic. When selecting more hydrocarbon groups having substituents can contribute additional polarity units of the formula (I). Therefore, it is preferable to choose RAaccordingly, the specific case of the methyl or ethyl, it is certainly substituted with halogen, -OH, -SH, -COOH, -NR'2, -COOR', -CONR', -CHO, where R' represents H or C1-3alkyl, particularly preferably, RAselected from methyl or ethyl.
In the units of the formula (II), RBis a hydrocarbon group, optionally substituted with halogen, -OH, -SH, -COOH, -NR2, -COOR", -CONR", -CHO, where R" represents hydrogen, alkyl or alkenyl, and RBis chosen so that the repeating element of the formula (II) are more hydrophobic than the repeating element of the formula (I). If R" is alkyl or aryl, it is preferred to C1-8alkyl or aryl group. If you have halogen substituents, it is preferable to choose Cl or F.
Preferably, as RBto choose C3-20hydrocarbon group, preferably, C3-12hydrocarbon group, more preferably, C3-6hydrocarbon group, and in particular, C4-6hydrocarbon group, all of which may be optionally substituted. However, more preferably, the hydrocarbon group had no Deputy.
As the hydrocarbon group is preferable aliphatic or aromatic groups such as alkyl groups, aryl groups or alkaline group. More preferred alkyl groups such as propyl, butyl, pentyl, hexyl, heptyl, octyl or nonyl, even more preferred C4-6alkyl g is uppy, i.e. butyl, pentyl, hexyl and especially preferred butylene groups, especially n-butyl.
The terms “hydrophilic” and “hydrophobic”as used in the context of this description, are widely used in this field. "Hydrophilic" refers to a preference for the water environment for the substance or its parts, i.e. hydrophilic substance or its part is easier to dissolve or be wetted by water than non-polar solvents, such as hydrocarbons. "Hydrophobic" refers to a preference for non-polar environment, i.e. a hydrophobic substance or a part of it easier to dissolve or wetted non-polar solvents, such as hydrocarbons, compared with water. The term "amphiphilic" refers to the simultaneous presence of hydrophilic and less hydrophilic or more hydrophobic parts in a substance that is often observed in surface-active substances. Thus, the copolymers used in the context of this invention also apply to the amphiphilic copolymers as include hydrophilic parts of the structure and less hydrophilic or more hydrophobic part, respectively.
If necessary, i.e. if the chemical structure RAand RBit is not obvious that a particular link of the formula (II) is more hydrophobic than a certain element of the formula (I), it can be set the way the comparison of the homopolymers of the respective links and definitions for them logP values in the same conditions. LogP is the logarithm of the partition coefficient observed for substances And between water and n-octanol. In particular, the separation factor for substance P And is defined as the ratio of the partition coefficient P=[A]n-octanol/[A]waterwhere [A] denotes the concentration of a substance in the appropriate phase. More hydrophilic substance will have a large concentration in the water. Usually, for measurements use the same amount of water and octanol.
In a preferred embodiment of the present invention, an appropriate choice of RBto obtain units of formula (II) c greater hydrophobicity in relation to the units of formula (I) can be checked by determining the critical concentration of mitselloobrazovaniya (ECR) copolymer containing structural data units, according to the method described in detail hereinafter. If CCM can be measured, the requirements for the hydrophilic properties of the units of formula (I) and more hydrophobic/less hydrophilic properties of the units of formula (II) duly executed.
Requirements units of the formula (I) to be hydrophilic, and with links in formulas (II) to be more hydrophobic in comparison to the units of formula (I) will also be appropriate made for any possible combination of preferred designs RAand RBthat will be clear of the respective structures. And the military, as RApreferably, choose a methyl or ethyl, optionally substituted by-OH, -SH, -COOH, -NR'2, -COOR', -CONR', -CHO, where R' may be H or C1-3acellam, and most preferably, be selected as RAmethyl or ethyl; and RBis selected from unsubstituted C3-20hydrocarbon group, preferably, C3-12hydrocarbon group, more preferably, C3-6hydrocarbon group, and in particular, C4-6hydrocarbon group, where the hydrocarbon groups are preferred aliphatic or aromatic groups such as alkyl groups, aryl groups or alkaline group. More preferred alkyl groups such as propyl, butyl, pentyl, hexyl, heptyl, octyl or nonyl, even more preferred, C4-6alkyl groups, i.e. butyl, pentyl, hexyl, and especially preferred, butylene group, in particular n-butyl.
It should be understood that under the water environment, water, aqueous solutions and the like in this application refers to the system of solvents, where 50% (volume ratio) or more, preferably 70% or more, more preferably 90% or more and, in particular, fully 100% of the total volume of the solvent(s) is water.
The copolymer containing the repeating unit of the formula (I) and (II)described above can be easily is prepared by polymerization with the opening of a series of 2-substituted-2-oxazolines (or 2-substituted-4,5-dihydrooxazolo in the IUPAC nomenclature). Thus, the polymers used in the context of this invention also apply to poly(2-oxazoline)am.
The copolymer according to this invention may include other repeating units in addition to the recurring units (I) and (II)above. However, it is preferable that a greater part of all repeating units, i.e. more than 50%, more preferably more than 75%, even more preferably greater than 90% and particularly preferably 100%, relative to the total number of the recurring units are recurring units of formula (I) or (II)as defined above. With all the repeating unit of the formula (II)contained in the copolymer will be more hydrophobic than any of the repeating units of the formula (I)contained in the copolymer.
The ratio of the number of repeating units of the formula (I) to the number of repeating units of the formula (II), generally ranges from 20:1 to 1:2, preferably from 10:1 to 1:1 and more preferably from 7:1 to 3:1.
Relative to the location of the repeating units (I) and (II)above, the copolymers according to the invention can be statistical copolymers, copolymers containing segments of polymerized units of one type (i.e. segments containing units of the formula (I), and/or segments containing units of the formula (II)), gradient copolymers or copolymers. B is exopolymer especially preferred.
The term "block copolymers" is used here in accordance with its established value in this field and relates to copolymers, where the repeating unit of a certain type are organized in blocks, i.e. the repeating unit of the same type sequentially polymerized directly one after the other, unlike, for example, sequences randomly alternating repeating units of different types. In other words, the blocks in blockcopolymer, such as blocks a and b, to be discussed below, are polymeric compounds themselves, obtained by polymerization of identical monomers or have similar characteristics.
Block copolymers to be used in accordance with the present invention comprises at least one block of poly(2-oxazoline)and consisting of repeating units of the formula (I)as defined above (which is hydrophilic due to the presence of relevant links) and at least one block of poly(2-oxazoline)and consisting of repeating units of the formula (II)as defined above (which is more hydrophobic than the block And, thanks to the presence of relevant links).
Block copolymers according to the invention may be referred to as amphiphilic copolymer due to the presence of at least one hydrophilic segment (block) and by at least one less hydrophilic or more hydrophobic segment (block).
Preferably, at least one block And blockcopolymer, more preferably, all of the blocks And if a lot of them, was(were) represented by formula (II):
where RApresents a methyl or ethyl group, preferably a methyl group, and n indicates the number of repeating units in the block A. Preferably, n is 5 or more, more preferably 10 or more, most preferably 20 or more. In most cases, n is less than 300, more preferably 200 or less, more preferably 100 or less, especially 50 or less.
Preferably, at least one block B of blockcopolymer, more preferably, all the blocks In, if a lot of them - was/were represented by structure (III):
where RB- C3-20hydrocarbon group, preferably, C3-12hydrocarbon group, more preferably, C3-6hydrocarbon group, and particularly preferably, C4-6hydrocarbon group. As hydrocarbon groups, preferred aliphatic or aromatic groups such as alkyl groups, aryl groups or alkaline group. More preferred alkyl groups such as propyl, butyl, pentyl, hexyl, heptyl, octyl or nonyl, even more preferred C4-6alkiline the group, i.e. butyl, pentyl, hexyl and especially preferred butylene groups, especially n-butyl. Variable n is preferably equal to 5 or more, more preferably 10 or more. In most cases, n is less than 300, more preferably 200 or less, or 100 or less, and particularly preferably 50 or less.
The block copolymers used in the context of this invention, the delivery system of medicinal product contains at least one block a and at least one block B, as described above. Also, it may contain one or more blocks other than A or B. However, it is preferable that the block copolymers contain only blocks a and B, which correspond to the above description. More preferably, all the recurring units of blockcopolymer was recurring units of the formula (I) or (II)described above.
Regarding the location of the blocks A and B in the copolymer used in the context of this invention, the preferred structure of the copolymer can be described as (AB)mor (BA)mwhere m is 1, 2 or 3, for example, ABA or BAB. More preferably, the block copolymers was AB or BA by diplomaprimary or ABA-tribocorrosion.
Thus, in the most preferred embodiment of the present invention, the polymer consists of a block(s), sostayashego is of polymerized 2-methyl-2-oxazoline or 2-ethyl-2-oxazoline (also referred to in the text as "poly(2-methyl-2-oxazoline)new blocks or poly(2-ethyl-2-oxazoline)new blocks) and block(s) B, consisting of polymerized 2-(C4-6alkyl)-2-oxazoline. Even more preferably, the copolymer consisted of a block(s) A, consisting of polymerized 2-methyl-2-oxazoline or 2-ethyl-2-oxazoline and block(s) B, consisting of polymerized 2-butyl-2-oxazoline (also called "poly(2-butyl-2-oxazoline)new block"). More preferred AB or ABA di - and triblocaltara with the above structure.
The copolymers used in the context of this invention typically have srednecenovogo molecular mass (Mn), measured by gel chromatography, in the range from 3 to 30, more preferably from 4 to 25 and most preferably from 6 to 20 kg/mol. The index of polydispersity (SDI=Mw/Mnwhere Mw- srednevekovaja molecular weight) is usually less than 1.3, preferably less than 1.25, and can even be set to 1,001.
It should be understood that the compositions also include combinations of, for example, a mixture or combination of two or more different types of copolymers are also described in this invention, for example, a combination of copolymers containing different groups of RAand/or RBor a combination of copolymers with different sequence of their repeating units, for example, a combination of statistical polymer and blockcopolymer.
The copolymers used in contact is kste of the present invention, can be prepared with known methods of polymerization. For example, poly(2-oxazoline)s can be obtained by cationic polymerization with the disclosure of the loop mechanism of living chains. Getting random copolymers, gradient copolymers and block copolymers are described, for example, R. Luxenhofer and R. Jordan, Macromolecules 39, 3509-3516 (2006), T. Bonné et al., Colloid. Polym. Sci., 282, 833-843 (2004) or T. Bonné et al. Macromol. Chem. Phys. 2008, 1402-1408, (2007).
With regard to the active substance (or active compound)contained in the compositions according to this invention, it should be understood that the compositions are usually convenient for the formulation of active substances which are poorly soluble in water, preferably, hydrophobic active substances or water-insoluble active substances. Therefore, the preferred active substances c solubility in water, for example, in demineralised water at 20°C less than 1 mg/ml, preferably less than 0.1 mg/ml, or even less than 0.01 mg/ml, and in particular, the solubility is less than 0.001 mg/ml Preferably limited solubility was observed in water in the pH range from 4 to 10.
Active ingredient (or active compound)used in the context of the present invention, preferably, is a biologically active substance (or biologically active connection is), including, but not limited to, substances used in the treatment (i.e. medication) or in the diagnosis, fungicides, insecticides, pesticides or herbicides and any other compounds used for protection of plants or crops, such as phytohormones, or active substances used in veterinary medicine. Used herein, the term "active substance" also includes compounds that are screened as potential leaders in the development of drugs or preparations for plant protection. Moreover, the present invention includes a method for the determination of active compounds interacting with the interests of the target, usually a biological target, such as a protein in a General sense, an enzyme or a receptor, a test screening method comprising the steps of introducing into the composition of the drug active compound according to the invention and conduct of the screening test, the obtained composition.
Specific examples of bioactive substances include, but are not limited to, drugs in the following categories: drugs acting at synaptic and neuroeffector nodal areas, medicines acting on the Central nervous system, drugs that affect inflammatory processes, medications that affect with the composition of body fluids, drugs affecting renal function and metabolism of electrolytes, cardiovascular drugs, drugs affecting gastrointestinal function, drugs affecting uterine contractility, chemotherapeutic substances for hyperproliferative diseases, in particular cancer chemotherapeutic substance for parasitic infections, chemotherapeutic substances for microbial diseases, antineoplastic agents, immunosuppressive agents, drugs acting on the blood-forming organs and degradation, hormones and their antagonists, dermatological drugs, antagonists of heavy metals, vitamins and nutrients, vaccines, oligonucleotides, and gene therapy. Specific medicinal substances which may be mentioned as suitable for use in the present invention include amphotericin b, nifedipine, griseofulvin, taxanes, including paclitaxel and docetaxel, doxorubicin, daunomycin, indomethacin, ibuprofen, etoposide, cyclosporine a, vitamin E and testosterone, in particular, paclitaxel, cyclosporine and amphotericin C. As mentioned above, the specific usefulness of this invention is shown for active substances which are insoluble in water.<> It is assumed that the compositions containing combinations, for example a mixture or combination of two or more active substances, such as two drugs also included in the invention.
It is generally preferable that the copolymer forms aggregates in the compositions in accordance with the invention, and more preferably that the resulting units such that the units of the copolymers include the active substance. Particularly preferred form of such aggregates is the micelle. Micelle, as mentioned here, in General terms, is an Assembly of amphiphilic copolymers with presents hydrophilic corona formed by the hydrophilic parts of the copolymer, and sequestered hydrophobic parts mentioned amphiphilic copolymer in the interior of the micelle. Particularly suitable copolymers for the formation of micelles are the copolymers described above as a preferred variant of the copolymers. Micelles mentioned in this invention are three-dimensional structures. Typically, micelles are formed when the concentration of amphiphilic molecules, elements in the aquatic environment exceeds a certain value. This value is known as the critical micellization concentration (CMC), which can be measured using a fluorescent probe, such as pyrene, which accumulates in g is grafalloy the core of the micelles, formed at concentrations above CMC. More specifically, micelles, according to this invention, are formed, for example, by self-organization of amphiphilic block copolymers in the hydrophilic, preferably water environment. In the formation of micelles hydrophilic region mentioned amphiphilic copolymers are in contact with surrounding solvent, while the hydrophobic region facing toward the center of the micelle. In the context of this invention, the center of the micelles typically comprises a hydrophobic active ingredient. The micelle can also be called "polymeric nanoparticle", due to its size, measured in nanometers, and because it is formed by a polymer.
Copolymers, especially block copolymers used in the context of this invention, generally have a low CMC values, typically less than 250 mg/ml In General, the CMC value varies between 5 and 150 or even 5 and 100 mg/L.
According to this invention, aggregates, especially micelles of different sizes can be generated pharmaceutical compositions according to the invention, depending on such factors as the molecular weight of the used copolymer or loading capacity of the medicinal product. Basically, it is preferable to use aggregates or micelles with a size in the range 5-500 nm. However, aggregates or micelles size 5-100 or 10-50 nm or even 10-30 n is, as shown by dynamic light scattering, can also be successfully formed, which is particularly useful for intravenous administration. It is noteworthy that the micelles usually have a narrow distribution of particle size (SDI ≤0.2 or even ≤0,1).
Usually aggregates and, in particular, the micelles formed in water or aquatic environment. Therefore, aggregates and, in particular, the micelle composition according to the invention, can be formed, for example, a method of dissolving thin films. In this method, the copolymer and the active substance are dissolved in a common solvent such as acetonitrile or dimethyl sulfoxide. After removal of the solvent (for example, in a stream of inert gas, mild heating and/or under the action of low pressure), films formed from the polymer and the active substance, can be easily dissolved in water or aqueous solvents and can be brought to the desired state with increasing temperature. After dissolution of the films is the formation of aggregates, mainly micelles. The stability of the formed aggregates allows drying the resulting solution to obtain a powder. For example, drying can be performed by lyophilization, usually without having to use createsite substances, with the possibility of further recovery in water or aqueous solvents without reducing the container of the particular boot or breakage of particles.
When using the above copolymers, the composition according to the invention typically form aggregates, soluble in water or in aqueous solutions, where they are stable for at least 12 hours at room temperature and higher temperatures, especially at temperatures below 40°C, which allows injecting the above-mentioned compositions to animals in General and man in particular.
The mass ratio of the current(is) of substance(s) to the copolymer(s) in the compositions according to the invention is usually 1:20 or higher, including 1:10 and above. Preferred weight ratio is at least 1:9, more preferably at least 2:8, even more preferably at least 3:7, and most preferably is 4:6. Basically, the ratio is 1:1 or less, if necessary, 8:10 or less.
Compositions according to the invention can be successfully received by active loading or drug loading (i.e. the ratio of the mass of the active substance or medicinal substance to the total weight of the active substance and blockcopolymer expressed as%) 10% or more, preferably 25% or more, more preferably 30% or more, more preferably 35% or more, particularly 40% or more. It is noteworthy that a sufficient solubility in water may be, to enom account obtained even for compositions according to the invention, with such high levels of drug loading even for active substances with a solubility of 10 mg/ml or even less than 5 μg/ml, such as paclitaxel. Thus, for example, compositions according to the invention allow solubilisate more than 7 mg/ml of paclitaxel, in particular 8 mg/ml or more, in water and aqueous solutions.
High capacity even for hydrophobic active substances coincides with unusual values obtained in the measurement of fluorescence spectra of pyrene in the pharmaceutical compositions according to the invention. The intensity ratio of peaks I1and I3in the fluorescence spectrum of pyrene is a measure of the polarity (K. Kalyanasundaram, J.K. Thomas, J. Am. Chem. Soc. 1977, 99, 2039-2044) environment paranavai samples. In water or similar polarity environment, this ratio typically varies between a 1.6 and 1.9 (K. W. Street, Jr., W. W. Acree, Jr. Analyst, 1986, 111, 1197-1201). In the presence of polymeric micelles pyrene becomes available less polar environment and attitude I1/I3usually decreases in parallel with the increase in fluorescence intensity. It is noteworthy that in the case of copolymers, especially block copolymers described herein, you may experience the opposite effect, when the ratio of I1/I3increases to values greater than 2, preferably 2.1 or even vyshe2,2, for example, to 2.35.
Due to the high efficiency of solubilization of the above-described copolymers and, in particular, block copolymers, compositions, in particular pharmaceutical compositions, in accordance with the invention are in the form of aqueous solutions, in principle it is sufficient if the amount of the copolymer ranges from as low concentration as 1 mg/ml, preferably 2 mg/ml to 100 mg/ml, preferably 50 mg/ml or 20 mg/ml As the copolymers are biocompatible, i.e. non-toxic and rapidly excreted in kidney cleansing the blood, high concentrations are not a limiting factor, but usually not required. This allows you greatly reduce the number of solubilization parenteral use of drugs, compared with hydrophobic drugs drugs available in the market, and, thus, reduce the risk of side effects.
In fact, the described block copolymers are able to reduce the required number of media for dissolution of paclitaxel about 100 and 9 times compared with Cremophor EL/ethanol (CrEL) and AbraxaneTM,respectively.
Moreover, the compositions of the described invention, the capacity (i.e. the amount of dissolved active substance/the number of source used active substance)*100%) can be equal to 100% and, as the government is about, very high (>80%). This is a major advantage as high efficiency load is very important for commercial applications to reduce production costs.
As described above, the copolymers according to this invention can be used to increase the solubility in water or aqueous solutions of active substances which are poorly soluble in water, predominantly hydrophobic active substances or water-insoluble active ingredients, and thus, they act as a solubilizer for data connections.
As a result, in a preferred embodiment of the present invention, the compositions of this invention also contain water in the form of an aqueous solution, emulsion or suspension, and in particular, it is preferable that they are aqueous solutions of the active substance and a copolymer. It should be understood that the term “solution” in the specific context includes colloidal solutions as they can form micelles in water. However, since the copolymers used in the context of this invention, allow the lyophilization compositions without reducing the activity and stability of the active substance and without the need to use cryoprotector, powders, especially freeze-dried powders, forming another preferred variant used in the various compositions according to the invention. These powders can be recovered in water or aqueous solutions.
Thus, the copolymers based on poly(2-oxazoline)Ah, described above, can be, for example, a universal delivery system with a high load for hydrophobic and structurally different drugs, such as paclitaxel, cyclosporine and amphotericin Century
Other forms of this invention are summarized in the following points:
1. Pharmaceutical composition consisting of a
(a) at least one biocompatible water-soluble amphiphilic blockcopolymer consisting of at least one block a and at least one block B, where a is a hydrophilic polymer selected from hydrophilic poly(2-oxazoline)s and B is selected from amphiphilic or hydrophobic poly(2-oxazoline)s and
(b) a hydrophobic bioactive compound, which forms aggregates, soluble in water or aqueous solutions, which are stable for at least 12 hours at room temperature and higher temperatures, especially at temperatures below 40°C, which provides parenteral application of the above-mentioned compositions for animals in General and humans in particular.
2. The pharmaceutical composition according to claim 1, where B represented by the following structural formula (III):
where RBthe hydrophobic side chain(containing saturated aliphatic chain, unsaturated aliphatic chain, saturated aliphatic ring or an unsaturated aliphatic ring or combinations thereof, and n is selected from 1 to 300.
3. The pharmaceutical composition according to any one of claims 1 or 2, where hydrophobic bioactive compound represented by the peptides, peptide, polyene, macrocycles, glycosides, terpenes, terpenoids, aliphatic and aromatic compounds and their derivatives, and other compounds having a solubility in water or aqueous medium at a pH in the range 4-10, equivalent to less than 1 mg/ml, preferably less than 100 μg/ml, even more preferably less than 50 μg/ml and most preferably less than 10 μg/ml.
4. The pharmaceutical composition according to any one of claims 1 to 3, where hydrophobic bioactive compound is selected from amphotericin B, nifedipine, grizeofulvina, paclitaxel, doxorubicin, daunomycin, indometacin, ibuprofen, etoposide and cyclosporine A.
5. The pharmaceutical composition according to any one of claims 1 to 3, where hydrophobic bioactive compound is paclitaxel.
6. The pharmaceutical composition according to any one of claims 1 to 5, where AB block copolymers are connected via a stable or unstable knitting for the formation of compounds which can be described as (AB)mwhere m ranges from 2 to 100, forming, for example, linear or star-shaped block copolymers, graft, Bloco aimery, dendrimers or hyperbranched block copolymers.
7. The pharmaceutical composition according to any one of claim 2 to 6, in which the hydrophobic side chain of R consists of 3-6 carbon atoms.
8. The pharmaceutical composition according to any one of claims 1 to 7, where amphiphilic block copolymers contains a block that is partially or completely consists of repeating units derived 2-butyl-2-oxazoline.
9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the hydrophilic polyoxazoles presents poly(2-methyl-2-oxazoline)Ohm or poly(2-ethyl-2-oxazoline)Ohm.
10. The pharmaceutical composition according to any one of claims 1 to 9, where soluble aggregates in the aquatic environment have a size of from 5 to 200 nm, preferably 10-100 nm.
11. The pharmaceutical composition according to any one of claims 1 to 10, comprising a hydrophobic bioactive compound and the amphiphilic block copolymers in a weight ratio of at least 1:9, preferably 2:8, more preferably 3:7, and most preferably 4:6.
Due to the inherent versatility of the pharmaceutical compositions according to the preferred embodiment of the present invention with the inclusion of bioactive substances/compounds, it should be understood that compositions suitable for the treatment or prevention of a wide range of diseases or disorders such as cancer, neurodegenerative diseases, hepatobiliary diseases, with techno cardiovascular disease and lung disease. The present invention also provides the use of the copolymers as defined above, for preparing a pharmaceutical composition for the treatment or prevention of any of these diseases. Moreover, the invention also provides diagnostic application of these compositions.
The term "cancer" in the context of this invention denotes a class of diseases or disorders characterized by uncontrolled cell division and the ability to spread, either by direct invasive germination in other tissues, either by introducing into tissue removed during metastasis (when migrating cancer cells in blood stream or lymphatic system).
The term "neurodegenerative disease" in the context of this invention denotes a class of diseases or disorders in which there is a destruction of neurons and in connection with the inability of the body to regenerate neurons (not counting a small number of neural stem cells), cells of the brain or spinal cord, for example, may not be fully restored. Related syndromes patients are ataxia and dementia.
The term "gastrointestinal and hepato-biliary disease" in the context of this invention denotes a class of diseases or disorders, acting on the liver, gall bladder and W is lonie ducts. Such diseases and disorders include, for example, cirrhosis, hepatitis, viral hepatitis, liver tumors, fatty metamorphosis of the liver, polycystic liver, Crohn's disease, ulcerative colitis, cholangiocarcinoma.
The term "cardiovascular disease" in the context of this invention denotes a class of diseases or disorders, acting on the heart and/or vessels.
The term "lung disease" in the context of this invention denotes a class of diseases acting on the respiratory system, which can be classified as obstructive, i.e. preventing the flow of air in and out of the lungs, and restrictive, i.e. reducing the functional capacity of lungs, States. Such diseases include, for example, asthma, bronchitis, asbestosis, fibrosis, sarcoidosis, lung cancer, pneumonia, pulmonary edema and pulmonary hypertension.
The pharmaceutical compositions according to this invention can be optionally compiled in conjunction with one or more pharmaceuticals excipients, such as carriers, diluents, fillers, disintegrating agents, lubricants, binders components, dyes, pigment substances, stabilizers, preservatives and/or antioxidants.
The pharmaceutical compositions can be prepared by technology, well known in this field such as technology, published in the 20th edition of Pharmaceutical Sciences, edited by Remington. The pharmaceutical compositions can be prepared for oral, parenteral, such as intramuscular, intravenous, subcutaneous, intradermal, intraarterial, rectal, topical pulmonary or vaginal administration. Dosage form for oral administration may be presented in the form of tablets, coated or not coated soft gelatin capsules, hard gelatin capsules, lozenges, pellets, solutions, emulsions, suspensions, syrups, elixirs, powders and granules to dissolve, dispersible powders and granules, medicinal chewing gums, chewable tablets and effervescent tablets. Dosage forms for parenteral administration include solutions, emulsions, suspensions, dispersions, and powders and granules to dissolve. Pharmaceutical form emulsion is preferred for parenteral use. Dosage forms for rectal and vaginal applications include candles and ovule. Dosage form for pulmonary/pulmonary delivery may be in the form of inhalation or insufflation, such as nebulizer metered doses. Dosage forms for topical application can be in the form of creams, gels, ointments, oils, PLA is tyra and transdermal delivery systems.
Introduction to the patient the pharmaceutical compositions according to this invention can be carried out in any convenient way, as systemically/perifericheskie and locally, including but not limited to, one or more of these, as orally (e.g. in the form of tablets, capsules or applied to the inside of the solutions), topically (e.g., transdermal, intranasal, ocular, transbukkalno or sublingual), parenteral (e.g., injection or infusion, including for example subcutaneous, cutaneous, intramuscular, intravenous, intraarterial, intracardial, intrathecal, vnutripolostnoe, vnutricapsulino, podkapsuliarnaya, intraorbitally, intraperitoneal, intratracheal, under the cuticle, intraarticular, subarachnoid, or epigastric injection using, for example, implant depot, for example, subcutaneously or intramuscularly), pulmonary (e.g. by inhalation or insufficie drug using, for example, an aerosol, e.g. through mouth or nose), gastrointestinal disorders, intrauterine, intraocular, subcutaneous, ophthalmic (including intravitreal or Mezcalero), rectal, and vaginal administration. Oral and parenteral, in particular intravenous administration in General preferable, because, according to the invention, the composition provides sufficient solubility and bioavailability for e is their methods of introduction, even when used hydrophobic active substances.
If pharmaceutical drugs are administered parenterally, then examples of such introductions include one or more of intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intrauterine, epigastric, intracranial, intramuscular or subcutaneous administration of the compounds, pharmaceutical compositions, and/or using the methods of infusion. In the case of parenteral administration, the composition should be used in the form of sterile aqueous solutions which may contain other substances, for example enough salts or glucose to make the solution was isotonic with respect to blood. These aqueous solutions may optionally be suitably buffered (preferably pH 3 to 9). The preparation of suitable parenteral drugs in sterile conditions can be readily accomplished by standard pharmaceutical techniques well known to the experts in this field.
Pharmaceutical compositions can also be administered orally in the form of tablets, capsules, oval, elixirs, solutions or suspensions, which may contain flavoring or color additives for immediate, delayed, modified, extended, intermittent or controlled release.
Table the TCI may include such media as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, Dooney calcium phosphate and glycine, dezintegriruetsja agents, such as starch (preferably corn, potato or cassava starch), sodium starch glycolate, sodium, croscarmellose and certain complex silicates, and connecting components for granulation, such as polyvinylpyrrolidone, hypromellose (HPMC), hydroxypropylcellulose (GOC), sucrose, gelatin and Arabic gum. In addition, lubricating components such as magnesium stearate, stearic acid, glyceryl begent and talc may be included. Solid compositions of a similar type may also be employed as fillers in gelatin capsules. Preferred carriers in this case include lactose, starch, cellulose, milk sugar or high molecular weight polyethylene glycols. In aqueous suspensions and/or elixirs, the active substance can be mixed with various sweeteners or flavors, color substances or dyes, with emulsifying and/or suspendresume substances, and also solvents such as water, ethanol, propylene glycol and glycerin, and combinations thereof.
Alternatively, the pharmaceutical compositions can be used in the form of suppositories or vaginal suppositories, or applied topically in the form of gels, hydro is oil, lotions, solutions, creams, ointments or powders. The compositions of this invention can also be applied dermal or transdermal, for example, when using skin patches.
The pharmaceutical compositions may, in particular, to enter pulmonary, or rectal route, and occulere. With the introduction of occulere, they can be formulated in the form microtrenching suspensions in isotonic saline solution with normalized pH or, preferably, in the form of isotonic solutions with normalized pH on the basis of physiological saline, optionally in combination with a preservative such as benzylamine chloride. On the other hand, the compositions can be formulated in an ointment such as petrolatum.
For local application on the skin, the pharmaceutical composition can be formulated in the form of ointments containing suspended or dissolved therein the active ingredient, for example, is mixed with one or more of the following substances: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, nonionic emulsified wax and water. On the other hand, they may be formulated as a special lotion or cream, suspended or dissolved, for example, mixtures of one or more of the following substances: mineral oil, servicemonitor, the polyethylene is ethylene glycol, liquid paraffin, Polysorbate 60, citylove esters wax, 2-octyldodecanol, benzyl alcohol and water.
Given the good solubilizing properties of compositions according to this invention, it should be understood that preferably, they should be entered in the form and/or in accordance with techniques that require dissolving the biologically active ingredient in the water.
It is assumed that in a typical scenario, the physician determines the dosage of the pharmaceutical compositions, the most relevant to a specific subject. The specific dosage and frequency of use in a particular subject may be varied and depends on a number of factors, including the disease or disorder to be treated or prophylaxis, type of bioactive compounds, the metabolic stability and the duration of the connection, the age, body weight, General health, sex, diet, mode and time of administration, rate of excretion of the drug from the body, use simultaneously with other drugs, the severity of the disease and the type passable subject therapy.
Suggested, but not limiting the dose of the compositions according to this invention for use in humans (having a weight of about 70 kg) may be from 0.1 μg to 10 g, preferably from 0.1 mg to 0.5 g, depending on the mass of the active substance (the. drug at standard dose. The standard dose can be used, for example, from 1 to 4 times a day. The dose will depend on the method of application. You should understand that you may need routine selection dosages, depending on the age and weight of the patient/subject, but also on the severity of the condition to be treated. Specific dosage and method of application should be assigned at the discretion of the attending physician or veterinarian.
Consider this invention subjects or patients in need of treatment or prevention, which introduces the compositions of this invention are to mammals. In addition to the treatment of humans, the invention provides for the treatment of economically and agronomically important mammals. Non-limiting examples of agronomically important animals include sheep, cattle and pigs, whereas, for example, cats and dogs can be considered as economically important animals. Preferably, the subject/patient man.
The term "treatment of a disorder or disease", used here, is widely distributed in this area. "Treatment of a disorder or disease" suggests that the disorder or the disease was diagnosed in the patient/subject. The patient/subject suffering from a disease or Russ the device, usually manifests specific clinical and/or pathological symptoms, which can be easily correlated with a specific pathological condition (i.e. diagnosed disease or disorder) specialist.
"The treatment of disorders or diseases may, for example, lead to stop the progression of the disorder or disease (i.e. not worsening symptoms or slow the progression of the disorder or disease (if delay in progression is only temporary). "The treatment of disorders or diseases may also lead to partial response (i.e. improvement of symptoms) or complete response (i.e. the disappearance of symptoms) in the subject/patient suffering from the disease or disorder. "Improvement" when the disorder or disease may, for example, to delay progression of the disease or disorder or a pause in the progression of the disease or disorder. For complete or partial response to treatment may be followed by relapse. It should be understood that the specific subject/patient may have a wide range of responses to treatment (such as listed above). Treatment of disorders or diseases may including to have a curative effect (preferably to result in a complete response and, ultimately, to recovery from illness or Zab is diseases) or palliative action (including the disappearance of symptoms).
Also, the term "prevention of disorder or disease" is used here, shirokoporistom in this area. For example, prevention of diseases or disorders may be useful for the patient/subject for which it is assumed predisposition to a disease or disorder as mentioned in the context. For example, the subject/patient may have a predisposition or susceptibility to a disease or disorder, including, but not limited to, hereditary predisposition. Such disposition can be determined by standard methods, using, for example, genetic markers or phenotypic indicators. It should be understood that the disorder or disease to be alert to this invention, have not been or cannot be diagnosed in a particular patient/subject (for example, the patient/subject may not show clinical or pathological symptoms). Thus, the "warning" implies the use of compounds according to this invention prior to the occurrence or diagnosis of any clinical and/or pathological symptoms or before they can be diagnosed or identified when visiting the doctor.
This description is quoted a large number of documents, including articles of magazines and p. the tents. The reported data, documents, information, although may not be considered patentable by the present invention, incorporated by reference in its entirety. More specifically, all cited documents are incorporated by reference to the same extent as if the quote each individual document specifically and individually indicated.
In more detail, the invention will be illustrated in the following examples. It should be noted that incorporated herein descriptions are not to limit the spread of the use of this invention.
1. General materials and methods
All substances for the preparation of polymers were purchased in Aldrich (Steinheim, Germany) and Acros (Geel, Belgium) and were used as received, unless specifically otherwise agreed. 2-Butyl-2-oxazoline (Buochs) were prepared as recently described (Huber, S. and Jordan, R., Colloid Polym. Sci. 286, 395-402 (2008)). Methyl triftormetilfullerenov (Meotf), 2-methyl-2-oxazoline (Meox), 2-ethyl-2-oxazoline (Etox), acetonitrile (atsn), and other solvents for the synthesis of the polymers were dried by heating at the reflux over San2in a dry nitrogen atmosphere and subsequent distillation before use. NMR spectra were obtained on Bruker Avance III 400, Bruker ARX 300 or Bruker AC 250 at room temperature. Spectra were calibrated, ISOE is isua the solvent signals (CDCl 3, 7,26 ppm, D2O 4,67 ppm). Helpanimals chromatography (GPC) was carried out on the system Waters (pump model 510, IR detector model 410, predalone and two Plgel PL Resipore column (3 µm, 300×7.5 mm) with N,N-dimethylacetamide (DMA) (75 mmol/l LiBr, 80°C, 1 ml/min) as eluent and calibrated against PMMA standards. Dynamic light scattering was performed on the instrument Zetasizer Nano-ZS (Malvern Instruments Inc., Southboro, Massachusetts) at room temperature.
2. Methods synthesis
Polymerization and methods selection of products was carried out in accordance with the previously described method (Luxenhofer, R. and Jordan, R., Macromolecules 39, 3509-3516 (2006); Bonné, T. B., et al., Colloid Polym. Sci. 282, 833-843 (2004)).
For example, obtaining methyl-P[Meeks27-b-Buochs12-b-Meox27]-piperidine (P1) was carried out as follows (here “Methyl” means that the polymer bears a terminal methyl group, “P” means that the part in brackets is a polymer part, “Meats” means a polymer units derived by polymerization of 2-methyl-2-oxazoline, the indices represent the number of repeating units in the respective polymer block, “b” indicates the beginning of a new block, “Buochs” means a polymer units derived by polymerization of 2-butyl-2-oxazoline and piperidine is another terminal group of the polymer chain):
In dry inert conditions to 32.2 mg (0.2 mmol, 1 equiv.) METI triftormetilfullerenov (methyl, triplet, Meotf) and 440 mg (5.17 mmol, 26 EQ.) 2-methyl-2-oxazoline (Meox) were dissolved in 3 ml of dry acetonitrile at room temperature. The mixture was subjected to microwave irradiation (150 W maximum, 130°C) for 15 minutes. After cooling to room temperature, the monomer of the second block, 2-butyl-2-oxazoline (256 mg, a 2.01 mmol, 10 EQ.) was added, and the mixture was irradiated in the same manner as in the first block. The procedure was repeated for the third block 442 mg (5,19 mmol, 26 EQ.). Finally, P1 was terminated by adding 0.1 ml of piperidine (1.01 mmol, 5 EQ.) at room temperature. After stirring overnight, was added To excess2CO3and the mixture is stirred for several hours. The solvent was removed after filtration and to the residue were added 3 ml of chloroform. After deposition of cold diethyl ether (approximately 10-fold amount with respect to the polymer solution), the product was isolated by centrifugation. The deposition was carried out in triplicate and the resulting polymer had the appearance colorless powder (792 mg, 67%, Mth=5.8 kg/mol) after lyophilization from water. GPC (DMA): Mn=8.5 kg/mol (SOPS 1,21);1H-NMR (CDCl3, 298 K): δ=3,45 (user, 255H, (N-CH2CH2)); 3.04 from/2,95 (m, 3H, N-CH3Ini); 2,43 is 1.86 (m, 212H, CO-CH3, CO-CH2CH2Pid ); 1,56 (user, 29H, CH2-CH2-CH2-); 1,32 (user, 28H, -CH2-CH3); of 0.91 ppm (user, 37H, -CH3butyl), Mn=6.2 kg/mol (Meox27-b-Buochs12-b-Meox27).
Obtaining methyl-P[Meox37-b-Buochs23-b-Meox37]piperidine (P2)
P2 was obtained accordingly, using 24 mg Meotf (0,146 mmol, 1 EQ), 333 mg Meox (3,91 mmol, 27 EQ, 1st block), 286 mg Buochs (2.25 mmol, 15 EQ, block 2) and 333 mg Meox (3,91 mmol, 27 EQ, 3rd block) and 80 μl of piperidine as the final reagent. The resulting product had the appearance colorless solid (795 mg, 83%, Mth=6.6 kg/mol).
GPC (DMA): Mn=10.4 kg/mol (SOPS 1,18);1H-NMR (CDCl3, 298 K): δ=3,44 (user, 360H, (N-CH2CH2)); 3,03/2,94 (m, 3H, N-CH3Ini); 2,33-1,9 (m, 279H, CO-CH3, CO-CH2CH2Pid); 1.55V (user, 47H, CH2-CH2-CH2-); 1,32 (user, 45H, -CH2-CH3); of 0.91 ppm (user, 68H, -CH3butyl), Mn=9.3 kg/mol (Meox37-b-Buochs23-b-Meox37).
Obtaining methyl-P[Meox36-b-Buochs30-b-Meox36]piperidine (P3)
P3 was obtained accordingly using to 24.7 mg of methyl triflate (0,150 mmol, 1 EQ.) and 334 mg of 2-methyl-2-oxazoline (3.9 mmol, 26 EQ., 1st block). An aliquot W mg (5% wt./wt.) the reaction mixture was selected for the analysis of the first block by means of NMR and GPC. The same was done after the second block (364,4 mg Buochs; 2,87 mmol, 20 EQ., 10% wt./wt. were selected for analysis). The third block (306,9 mg Meox; 3.6 mmol, 28 EQ.) was added, the polymerization was stopped by adding 80 μl of piperidine and the obtained product had the appearance colorless solid (598 mg, 65%, Mth=6.6 kg/mol).
GPC (DMA): Mn=9.9 kg/mol (SOPS 1,23);1H-NMR (CDCl3, 298 K): δ=3,45 (user, 405H, (N-CH2CH2)); 3,03/2,95 (m, 3H, N-CH3Ini); 2,43 is 1.86 (m, 329H, CO-CH3, CO-CH2CH2Pid); 1,57 (user, 63H, CH2-CH2-CH2-); 1,32 (user, 60H, -CH2-CH3); of 0.91 ppm (user, 88H, -CH3butyl), Mn=10,0 kg/mol (Meox36-b-Buochs30-b-Meox36).
Obtaining methyl-P[Etox50-b-Buochs19]piperazine (P4)
P4 was obtained accordingly from 10 mg Meotf (61 μmol, 1 EQ.), 321 mg of 2-ethyl-2-oxazoline (Etox, 3,24 mmol, 53 EQ., 1st block) and 157 mg Buochs (1,23 mmol, 20 EQ., 2nd unit)using 150 mg of piperazine as the final agent. For deposition was used a solvent mixture of cyclohexane and diethyl ether (50/50 by volume). The resulting product was in the form of a colorless solid (yield 0.36 g, 77%, Mth=7.8 kg/mol).
GPC (DMA): Mn=11.5 kg/mol (SOPS 1,09);1H-NMR (CDCl3, 298K): δ=3,45 (user, 276H, (N-CH2CH2)); 3.04 from/2,95 (m, 3H, N-CH3Ini); 2,5-2,2 (m, 144H, CO-CH2-CH3, CO-CH2CH2Pid); 1,58 (user, 37H, CH2-CH2-CH2-); 1,34 (user, 41H, -CH2-CH3); 1,11 (user, 151H, CO-CH2-CH3); of 0.91 ppm (user, 56H, -CH3butyl), Mn=7.5 kg/mol (ATOX50-b-Buochs19).
3. Measurement of the fluorescence of pyrene
CMC was determined by a standard method. Briefly, a solution of pyrene in acetone (2.5 mm) was added to the vials and the solvent evaporated. The polymer solutions of the respective concentrations in the experimental solution were added to the same vial so that the final concentration of pyrene was 5×10-7M. the Solutions were incubated at 25°C (>2 h) and fluorescence spectrum of pyrene was measured on Fluorolog3 (HoribaJobinYvon) when λvasb=333 nm, λCOI=360-400 nm, slit width (vosb) = slit width (COI) = 1 nm step width dimension of 0.5 nm. Usually were averaged 5 spectra for each value (integration time of 0.1 sec, if you were averaged 10 spectra with 0.2 sec integration), CMC is defined as the concentration at which there is a rapid increase in fluorescence intensity. Moreover, the intensity of the fluorescence bands I1compared with the intensity of the band I3that allows to estimate arnosti environment paranavai samples. The formation of dimers in the electronically excited state has not been detected.
Using this approach, we established a low critical concentration of mitselloobrazovaniya (ECR)ranges from 100 mg/l (15 μm, P1), 20 mg/l (2.7 μm, P2), 7 mg/l (1 μm, P3) to 6 mg/l (0,7 µm, P4), respectively (see figure 1).
Figure 1 shows the intensity of fluorescence and the relationships I1/I3solutions of pyrene (5×10-7M in phosphate-buffered saline) as a function of the concentration of the copolymers used in the context of the invention, at 25°C.
4. Studies of solubilization of drugs
4.1 Solubilization of paclitaxel (PTX)
Solutions of polymers with drug were prepared by the method of thin films. The appropriate amount of polymer and paclitaxel (Sigma-Aldrich, St. Louis, Missouri, the order number T)(original solution 5-8 mg/ml in acetonitrile or ethanol) were dissolved in a minimum amount of acetonitrile or ethanol, respectively. After complete removal of solvent, the solid film of polymer and drug was soaked in assay buffer or deionized water. A typical procedure is as follows:
The solvent was removed in a stream of air or low heat and the films were exposed to 0.2 mbar for at least 3 hours to remove traces of RA is the solvent. Then 200 µl of assay buffer (an aqueous solution containing 122 mm NaCl, 25 mm Na2CO3, 10 mm HEPES, 10 mm glucose, 3 mm KCl, 1.4 mm CaCl2and 0.4 mm K2HPO4pH=7,4) were added to obtain a final concentration of the polymer. At higher concentrations of paclitaxel dissolution was facilitated by incubation of the solution at 50-60°C, usually within 5-10 minutes. Transparent solutions were filtered through HPLC filter for syringes (pore size 0.45 μm) and subjected to HPLC analysis (see below). Replacement of relatively toxic acetonitrile more inert ethanol as a common solvent to form a film without lowering the efficiency of loading.
An attempt was made to dissolve 4, 7 and 10 mg/ml of paclitaxel 10 mg/ml P2. Up to concentrations of paclitaxel 7 mg/ml were obtained transparent solution after a short weak heating (approximately 40°C). Under these conditions, the solubilization of paclitaxel was complete as confirmed by HPLC (see figa).
Only when the concentration of paclitaxel 10 mg/ml some transparent crystals remained undissolved even after 30 minutes of heating at 60°C. however, The unusually large amounts of paclitaxel (8.2 mg/ml) were dissolved and was found in the aqueous phase, in other words, the resulting preparation is at least 40% wt. from Pak is Taxila. Similar results were obtained with other polymers, including P1, which has only 12 units in the block Buochs (see figv).
Even with such a low concentration of the polymer as a 2 mg/ml were obtained great download efficiency and the total loading of the drug in 30% wt. (see figs, D). Thus, the copolymers used in this invention can reduce the number of media required for dissolution of paclitaxel in about 100 and 9 times compared with Cremophor EL/ethanol (CrEL) and AbraxaneTMrespectively.
Figure 2 presents the results of solubilization of paclitaxel with amphiphilic copolymers using the method of thin films A-D). The concentration of paclitaxel in solution (the columns of the histogram) and the efficiency of load (crossed circles), obtained by using different polymers and the target concentration of paclitaxel: A) P2 (10 mg/ml) and 4 mg/ml, 7 mg/ml and 10 mg/ml of paclitaxel; (B) P1-P4 (10 mg/ml) and 4 mg/ml of paclitaxel; (C) P3 (2 mg/ml) and 100 μg/ml, 500 μg/ml and 1 mg/ml of paclitaxel; (D) P1-P3 (2 mg/ml) and 500 μg/ml of paclitaxel.
Data are presented as averages (n=3; in C: 1 mg/ml of paclitaxel n=1 and B: P4 n=2) ± standard deviation.
4.2 Solubilization of cyclosporin a
Solubilization of cyclosporin A (Alexis Corporation, San Diego, California, order number 380-002-G001) was carried out accordingly by the method of thin captured the K. Using P2 and cyclosporine a were obtained transparent and stable solutions.
4.3 Solubilization of amphotericin B
Solubilization of amphotericin b using P2 was carried out according to the method of replacement of the solvent by dialysis. P2 (10,2 mg) and trihydrate amphotericin B (2.1 mg, Riedel-de Haën, Salts, Germany, the order number 46006) were dissolved in 250 μl of dimethyl sulfoxide (DMSO)to obtain a transparent yellow solution. Just was added 750 μl of deionized water, after 100 μl of the solution were dull. The resulting mixture was placed in a dialysis bag (MWCO 3500 g/mol). The solution deliberately against 2 l of deionized water (the water was changed after 2, 4 and 22 hours). After a total of 50 hours, the suspension (4 ml) was collected from the package. An aliquot of 500 μl was filtered (0.45 µm)to get rid of particles and a transparent yellow solution was freeze-dried to obtain 1 mg yellow foamy solid. The residue was dissolved in 200 μl DMSO, and the number of amphotericin b was determined spectrophotometrically by measuring the absorbance at 410 nm. Validirovannyj the solution contained 366 µg of amphotericin b (18% wt./wt., with respect to P2). Another aliquot of 1 ml was dried (2.2 mg yellow foam) and subsequently dissolved in 100 μl of deionized water. Foam polymer and drug quickly disappeared completely with the formation of Yar what about the yellow solution with low viscosity. Thus, 3.7 mg/ml amphotericin B can be solubilisation using only 18.3 mg/ml P2 without using closewith substances. Using the same method, the water solubility of amphotericin B was determined as equal to about 0.4 mg/ml
4.4 HPLC analysis of solubilization of drugs
HPLC was carried out at isocratic conditions on the system Shimadzu, including SCL-10A system controller, SIL-10A autoinjector, SPD-10AV UV detector and 2 LC-10 AT the pump. Nucleosil C18-5 ál column (250 mm × 4 mm) was used as stationary phase, a mixture of acetonitrile/water (55/45 by volume) was used as the mobile phase. Detection was performed at 220 nm. The amount of paclitaxel in the polymer solution was calculated from a calibration curve obtained using known concentrations of paclitaxel dissolved in acetonitrile, and analyzed accordingly.
HPLC solutions cyclosporine a, obtained by the method described above was carried out using a mixture of acetonitrile/water (90/10 by volume) as mobile phase at 70°C. In a 5 mg/ml P2 was dissolved 1,03 mg/ml cyclosporine A. This corresponds to 82% efficiency loading and 17% loading (wt./wt.). Thus, using P2 managed to get about 120-fold increase in solubility of cyclosporine A.
4.5 Characterization of micelles
Loaded medicament is i.i.d. substance micelles were very small (radius 12-22 nm) and showed a narrow size distribution (polydispersity ≈ 0,04-0,12), measured by dynamic light scattering. Figure 3 presents the results of measurements of dynamic light scattering loaded drug micelles P1 and P2 (10 mg/ml), in both cases, containing 4 mg/ml of paclitaxel.
5. Method for the determination of cytotoxicity; analysis based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)
Cell line MCF7/ADR (derived from breast cancer cells human MCF7 (ATCC HT-B22) by selection with doxorubicin) were kindly provided by Y. L. Lee (William Beaumont Hospital, Royal Oak, pieces of Michigan). Cells were cultured in the medium Needle in the modification of Dulbecco (DMEM) supplemented with 10% teploizolirovannye fetal bovine serum (FBS) and 1% penicillin/streptomycin, as described in other sources. The whole environment for cell culture were purchased from Gibco Life Technologies Inc. (Grand island, new York).
Cells MCF7/ADR were planted in 96-well plates (104cells per well) and left to attach for 24 hours. Existing solutions were prepared from a 1 mg/ml initial solution of the polymer in the working buffer (containing 122 mm NaCl, 25 mm NaHCO3, 10 mm glucose, 10 mm HEPES, 3 mm KCl, 1.2 mm MgSO4, 1.4 mm CaCl2and 0.4 mm K2HPO4, pH 7.4) by dilution in an appropriate amount of environment (Wednesday Needle in the modification of Dulbecco (DMEM) supplemented with 10% FBS, 25 mm HEPES and 1% penicil is in/streptomycin). Cells were incubated 48 hours in 200 μl of the current solution. After removal of the incumbent solution, the cells are washed three times with phosphate-saline buffer (FSB). Then the cells were added to DMEM containing no FBS (100 μl/well), and 25 μl of 5 mg/ml solution of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Invintrogen, Eugene, PCs Oregon) in the FSB, and incubated for 2 hours at 37°C. Then the medium was removed and added to 100 μl of solvent (25% wt./wt. DMF, 20% wt./the amount of LTOs in water). Purple formosanus product was dissolved during the night, after which we measured the absorbance at 570 nm on a tablet reader (SpectraMax M5, Molecular devices). Positive controls were cells incubated only with the environment; negative controls were wells without cells. Each concentration was measured in 4 repetitions, data are presented as mean ± standard deviation.
Figure 4 presents the results of the cytotoxicity of the polymers P1-P4 in cells with multidrug-resistant (MCF7/ADR, A) and not to be resistant (MCF7) cells of the human adenocarcinoma cells and Madine-Darby kidney of the dog (MDCK) after 24-hour and 2-hour incubation (b, C). Data are presented as mean ± standard deviation (n=4). P1-P4 one was nectocarcinus at concentrations up to 20 mg/ml at 24-hour incubation in different cell lines.
In epaves one polymer, micelles loaded with paclitaxel, have a significant concentration-dependent toxicity in human and murine cell lines tumor (figa). For example, after 24 hours of incubation with micelles P2, P3, and P4, loaded with paclitaxel, the concentrations required to inhibit cell growth by 50%, IC50in resistant cells (MCF7/ADR) were within 10 microns. Commercially available drug CrEL-PTX was used as the control and comparative caused suppression of cell growth. Micelles loaded with paclitaxel, can be dried without the use closewith agents and can be easily resuspendable in water or physiological solution, with no harm to the loading of drug substance particle size or efficacy of a drug in vitro (pigv).
Figure 5 presents the dependence of the viability of the cells with multidrug-resistant MCF7/ADR, the concentration of paclitaxel. (A) Comparison of drugs paclitaxel with P2 and P3 indicates no significant difference in cell viability depending on the material of the medium after 24-hour incubation. B) For example, P4 is shown that changes in the activity of paclitaxel after lyophilization and resuspendable deionized water is not detected. Data are presented as ednie ± standard deviation (n=3).
6. Experiments with animals
All experiments were performed on female S/BI/6 mice at the age of 11-12 weeks, (Taconic Laboratories, Germantown, new York). The animals were kept at 5 in the cell with a lid and filter the air in a room with controlled light cycle (12-hour cycle of light/dark) and temperature (22F1 8C). All procedures with animals were carried out in sterile kelp. Food and water were given unlimited. The animals were kept in accordance with the Principles of the Animals that are defined by the National Institute of health, and the protocols were approved by the institutional Committee of the maintenance and use of animal Medical center of the University of Nebraska. Cell lung carcinoma Lewis (LLC 3T) were grown in Kzt75 vessels and was going through a balanced salt solution Hanks. The cell suspension (1×106/mouse) were injected subcutaneously in 50 μl into the right side. After tumors had formed, they were measured (day 1) and existing solutions with a concentration of paclitaxel 10 mg/ml was administered intravenously on days 1, 4 and 7.
In vivo antitumor effect of micelles loaded with paclitaxel was studied in C57/BI/6 mice with subcutaneous tumors lung carcinoma Lewis (6). Commercially available CrEL, as well as restored preparations of poly(2-oxazoline)and paclitaxel (P2-station reserve fuel facility) significantly (p<0,05) decrease is or the severity of the tumor after a single injection (day 4, inhibition of tumor growth 72% and 63%, respectively). Tumors in mice which were injected P2-station reserve fuel facility remained significantly lower (p<0.05) than in mice, which was introduced commercially product between days 11 and 25. We found that suppression of tumor growth drug P2-station reserve fuel facility during this period was approximately 70% compared to 50-60% in the group of mice that were injected CrEL. However, after 28 days, there was a sharp deterioration in the severity of tumors in animals in group P2-station reserve fuel facility and showed similar inhibition of tumor growth in both treatment groups.
Figure 6 (A) presents the comparison of the relative mass of subcutaneous tumors lung carcinoma Lewis in C57/BI/6 mice in the negative control (saline only P2)mice, which were introduced paclitaxel, the solubilized in poly(2-oxazoline)s (P2-station reserve fuel facility) and a commercial product CrEL at the same doses station reserve fuel facility (10 mg/kg). Arrows indicate the time of injection. In Suppression of tumor growth P2, P2-station reserve fuel facility and a commercial product CrEL. Data are presented as mean ± standard deviation (n=5). Figure 6) presents the calculated suppression of tumor growth in the treatment group P2, P2-station reserve fuel facility and CrEL at different points in time.
1. Composition for delivery of active ingredient containing:
(a) at least one block copolymers containing at least one poly(2-oxasolinone)block a, consisting of autoraise units of the formula (I)
where RAmeans a hydrocarbon group which optionally may be substituted by-OH, -SH, -COOH, -NR'2, -COOR', -CONR', -CHO, where R' represents H or C1-3alkyl, and RAis chosen so that the repeating element of the formula (I) is hydrophilic;
and at least one poly(2-oxasolinone) block consisting of recurring units of the formula (II),
where RBmeans a hydrocarbon group which optionally may be substituted with halogen, -OH, -SH, -COOH, -NR2, -COOR", -CONR", -CHO, where R ' denotes H, alkyl or alkenyl, and RBis chosen so that the repeating element of the formula (II) are more hydrophobic than the repeating element of the formula (I); and
(b) one or more active substance.
2. The composition according to claim 1, where RAchosen independently in each case, from methyl and ethyl, optionally substituted by-OH, -SH, -COOH, -NR'2, -COOR', -CONR', -CHO, where R' represents H or C1-3alkyl, and RBselected from C3-20hydrocarbon group.
3. The composition according to claim 1, where RAchosen independently in each case from methyl and ethyl, a RBselected from C4-6the alkyl.
4. The composition according to claim 1, where at least one block a is a block of poly(2-methyl-2-oxazoline) or poly(2-ethyl-2-oxazoline) and at least one block is a block of poly(2-b the Teal-2-oxazoline).
5. The composition according to claim 1, where block copolymers is AB or BA by diplomaprimary or ABA or BAB-tribocorrosion.
6. The composition according to claim 1, where at least one active substance is a hydrophobic active ingredient.
7. The composition according to claim 1, where at least one copolymer forms micelles that can include the active substance.
8. The composition according to claim 1, where the mass ratio of active substance (substances) to the copolymer (copolymer) is from 1:10 to 1:1.
9. The composition according to claim 7, where the micelles have a size of from 5 to 500 nm.
10. The composition according to claim 1, where the active load, expressed as the ratio of the mass of the active substance or medicinal product to the total weight of the active substance and blockcopolymer equal to 10% or more.
11. Composition according to any one of claims 1 to 10, which is a pharmaceutical or diagnostic composition comprising a medicinal substance as an active substance.
12. The use of a copolymer as defined in any one of claims 1 to 5 to solubilize the active substance in water or aqueous solution.
SUBSTANCE: invention relates to a mixture of graft copolymers for use as an additive in chemical materials, as well as in development, exploitation and integration of underground deposits of oil and natural gas and in case of deep wells. The mixture of graft copolymers contains at components, at least one representative of brown coal, brown coal coke, lignite and a brown coal derivative, at least one representative of natural polyamides and different vinyl-containing compounds. The grafting base is selected from at least one representative of brown coal, brown coal coke, lignite and a brown coal derivative such as tannin and/or such a polyphenol derivative as lignosulphonate, or a polyamide component. Suitable polyamide components are natural polyamides, preferably caseins, gelatins and collagens, bone glues, blood albumins, soya proteins and products of splitting thereof, which are formed via oxidation, hydrolysis or depolymerisation, as well as mixtures thereof. The grafting component used is representatives of brown coal, brown coal coke, lignite, a brown coal derivative and natural polyamides, as well as vinyl-containing compounds in their O-, S-, P- and N-forms and styrenes, which can be in sulphonated form, or as a graft product. The graft product is obtained by grafting a vinyl-containing compound to at least one representative of natural polyamides or mixtures thereof, or grafting a vinyl-containing compound to at least one representative of brown coal, brown coal coke, lignite and a brown coal derivative. Graft copolymers with preferred molecular weight are used as a mixture, particularly in chemical construction materials and during development, exploitation and integration of underground deposits of oil and natural gas and in case of deep wells or as an additive for compositions containing hydraulic binding substances, as a water-retaining agent and/or liquefier.
EFFECT: graft copolymers have excellent resistance to salt and temperature and are also water-soluble and/or biodegradable.
17 cl, 2 tbl, 7 ex
SUBSTANCE: described is a method of producing a highly purified disinfectant which contains polyhexamethylene guanidine hydrochloride, characterised by that flush water from production of polyhexamethylene guanidine hydrochloride in solid form is used. The flush water is mixed with 25% NaCl solution in ratio of 1:1 (polyhexamethylene guanidine hydrochloride solution with impurities: NaCl solution) while stirring constantly for 1 hour at temperature in the range of 50-60°C. Stirring is stopped and the mixture is cooled to 5°C, thereby dividing the mixture into two parts; the bottom part is drained, neutralised and sent for recycling and the top part containing up to 60-70% polyhexamethylene guanidine hydrochloride is further diluted to 50% with an aqueous solution of quaternary ammonium salts (QAS) to QAS concentration of not higher than 5%.
EFFECT: extracting highly purified, concentrated high-molecular weight polyhexamethylene guanidine hydrochloride from flush water, reducing the amount and toxicity of waste water, improving disinfecting properties and increasing output of the commercial-grade product when producing polyhexamethylene guanidine hydrochloride in solid form, reducing cost.
1 cl, 1 ex, 2 tbl
SUBSTANCE: method of producing a disinfectant involves first carrying out polycondensation of hexamethylenediamine and guanidine hydrochloride. Polycondensation starts with preparation of a reaction mass in form of a suspension of crystalline guanidine hydrochloride in molten hexamethylenediamine, taken in ratio of 1:(1-1.5). The suspension is obtained by gradually adding crystalline guanidine hydrochloride, preheated to temperature of 90-120°C, to molten hexamethylenediamine and then stirring. The reaction mass is then heated in steps: holding for 4 hours at 120°C, then for 8 hours at 160°C and then for 3 hours at 180°C. Temperature is then gradually raised to 210°C at a rate of 3-4°C/h. The reaction mass is then subjected to vacuum treatment and cooled.
EFFECT: method enables to reduce toxicity of the end product and obtain a polymer with the required molecular weight and sufficient purity without washing steps.
SUBSTANCE: described is a method of producing polyguanidines by polycondensation of a guanidine salt with a diamine while heating, characterised by that polycondensation is carried out in the presence of an organic acid or a mixture of organic acids and heating is carried out in steps as follows: at the first step at 120-130°C for 0.5-1 hour; at the second step at 150-160°C for 3.5-4 hours; at the third step at 170-180°C for 0.5-1.5 hours.
EFFECT: improved method.
11 cl, 2 tbl, 4 ex
SUBSTANCE: present invention relates to polyphenylene ether ketone oximates, as well as a method for production thereof. An elementary unit of said polyphenylene ether ketone oximate has the formula: [-O-N=C(CH3)-C6H4-O-C6H4-C(CH3)=N-O-C6H4-CO-C6H4]n. The polyphenylene ether ketone oximate has reduced viscosity of 0.4-0.5 dl/g and molecular weight ranging from 40800 to 51000. Polyphenylene ether ketone oximate are obtained by non-equilibrium nucleophilic polycondensation of difluorodiphenyl ketone with diacetyl diphenyl oxide diketoxime. The reaction is carried out in dimethyl sulphoxide for 6 hours at 165°C, with reaction of equimolar amounts of potassium diacetyl diphenyl oxide dioximate with 4,4'-difluorobenzophenone. The molar ratio 4,4'-difluorobenzophenone: 4,4'-diacetyl diphenyl oxide diketoxime: KOH: K2CO3 is equal to 1:1:2:0.15.
EFFECT: obtained polymer has improved mechanical properties, heat-resistance, and also has a system of properties which are characteristic for both polyether formal oximates and polyether ketones.
2 cl, 3 ex
SUBSTANCE: present invention relates to a method of producing oligo-3,3-bis(azidomethyl)oxetane which is used as a hydroxyl-containing compound for producing energy-intensive polyurethane thermoplastic elastomers. The method involves cationic polymerisation of 3,3-bis(chloromethyl)oxetane in methylene chloride at 20-35°C in the presence of 1-10 wt % boron trifluoride etherate and a diatomic alcohol in molar ratio of 1:(5-15) to 3,3-bis(chloromethyl)oxetane. Further, the intermediate oligo-3,3-bis(chloromethyl)oxetane is separated in finely dispersed form, for which at the end of polymerisation, an organic solvent is added to the reaction mass, methylene chloride is evaporated and oligo-3,3-bis(chloromethyl)oxetane is precipitated with water. Sodium azide is then added to the obtained finely dispersed oligo-3,3-bis(chloromethyl)oxetane in a medium of an organic solvent at 90-130°C in the presence of 0.5-3 wt % tetrabutylammonium bromide.
EFFECT: highly efficient method of producing oligo-3,3-bis(azidomethyl)oxetane and high output of the end product.
5 cl, 1 tbl, 7 ex
SUBSTANCE: disclosed are oligomers based on hexamethylene guanidine derivatives of formula (I), where R denotes or , n1, n2 and n3 equal 1-3, z equals 0.15-1.10; with molecular weight distribution Mw/Mn from 5.4 to 9.3, with weight-average molecular weight Mw 3800-6300 and number-average molecular weight Mn 600-1100. Disclosed also is a disinfectant containing disclosed oligomers as an active component, as well as use thereof.
EFFECT: disclosed compounds have improved and steadily reproducible disinfectant properties, low toxicity and corrosiveness.
4 cl, 1 dwg, 6 tbl, 8 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: inventions relate to field of medicine, veterinary, agriculture and pharmacology and deals with biocidal polyguanidine of formula Acid or (1), method of its obtaining and based on it biocidal composition.
EFFECT: inventions make it possible to obtain biocidal polyguanidine with high antimicrobial activity with wide spectrum of action.
4 cl, 14 tbl, 17 ex
SUBSTANCE: invention relates to chemistry and physics-chemistry of polymers, specifically to novel copolymers of N-vinylcarbazole and N-vinylcaprolactam. The invention also relates to solid solutions of polymethyl methacrylate which contain Tb3+ ions and said copolymers. Described are copolymers of N-vinylcarbazole with N-vinylcaprolactam and solid solutions of polymethyl methacrylate containing Tb3+ ions and copolymers of N-vinylcarbazole with N-vinylcaprolactam.
EFFECT: obtaining metal-polymer solid solutions in polymethyl methacrylate based on copolymers of N-vinylcarbazole with N-vinylcaprolactam of the disclosed composition, characterised by high luminescence intensity between 344 and 700 arbitrary units, and the obtained copolymer solid solutions can be of interest in the design of laser, luminescent, electroluminescent ad high-speed switching devices.
2 cl, 8 ex, 1 tbl, 3 dwg
SUBSTANCE: invention relates to chemistry of high-molecular compounds and specifically to a poly-3,3-bis(azidomethyl)oxetane synthesis method. Poly-3,3-bis(azidomethyl)oxetane is obtained by reacting poly-3,3-bis(chloromethyl)oxetane with sodium azide in a dimethylformamide medium. The poly-3,3-bis(azidomethyl)oxetane is obtained in finely dispersed form which is easy to apply.
EFFECT: poly-3,3-bis(azidomethyl)oxetane synthesis method enables to reduce the amount of dimethylformamide used and also enables to avoid use of methanol.
1 cl, 18 ex, 1 tbl
SUBSTANCE: method of producing copolymers of polyguanidine salts involves treating polyguanidine chloride with a metal compound in an alcoholic solvent, separating the formed metal chloride precipitate through filtration, mixing the filtrate and acid with formation of a copolymer of polyguanidine salts, and subsequent drying the desired product. The polyguanidine chlorides used have general formula , where R is alkylene with straight and branched structure with C4-C12, oxaalkykene[-CH2-]P O[-CH2-]q, dioxaalkylene [-CH2-]P O[-CH2-]q O[-CH2-]r, trioxaalkylene [-CH2-]pO[-CH2-]q O[-CH2-]r O[-CH2-]s, tetraoxaspiroalkylene , n ranges from 2 to 1000; p, q, r, s assume values from 2 to 12, the metal compound used is potassium hydroxide, the alcoholic solvent used is a mixture of ethanol and rectificate with isopropanol in ratio (100-30):(0-70), where the said alcoholic solution of potassium hydroxide is mixed with dry crushed polyguanidine chloride, mixture of organic or mineral (except hydrofluoric) mono- or polybasic acid or mixture acid with filtrate is done after vacuum distillation of alcohol from the filtrate at pressure ranging from 0.1 to 0.2 atm and temperature from 50 to 60°C. Copolymers of polyguanidine salts of general formula: , where R is alkylene with straight or branched structure with C4-C12, oxaalkylene [-CH2-]P O[-CH2-]q, dioxaalkylene [-CH2-]p O[-CH2-]q O[-CH2-]r, trioxaalkylene [-CH2-]P O[-CH2-]q O[-CH2-]r O[-CH2-]s, tetraoxaspiroalkylene, where (n-x): x=19:1-1:19; n=2-100; p, q, r, s assume values from 2 to 12, A is an anion of organic or mineral (except hydrofluoric) mono- and polybasic acid or a mixture of acids.
EFFECT: improvement of method.
3 cl, 4 ex, 4 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: in formula R1 is H or (1-6C alkyl); R2 represents NRbRc, (1-4C)alkyl, (1-4C)fluoroalkyl, CF3, (1-4C)hydroxyalkyl, -(1-4Calkyl)hetAr1, -(1-4Calkyl)NH2, -(1-4C alkyl)NH(1-4Calkyl), -(1-4Calkyl)N(1-4Calkyl)2, hetAr2, hetCyc1, hetCyc2, phenyl substituted where applicable by NHSO2(1-4Calkyl) or (3-6C)cycloalkyl, substituted where applicable by (1-4C alkyl), CN, OH, OMe, NH2, NHMe, N(CH3)2, F, CF3, CO2(1-4C alkyl), CO2H; C(=O)NReRf or C(=O)ORg; Rb is H or (1-6C alkyl); Rc represents H, (1-4C)alkyl, (1-4C)hydroxyalkyl, hetAr3 or phenyl, wherein the above phenyl is substituted where applicable by one or more substitutes independently from halogen, CN, CF3 and -O(1-4C alkyl); Re represents H or (1-4C)alkyl; Rf represents H, (1-4C)alkyl or (3-6C)cycloalkyl; Rg represents H or (1-6C)alkyl; X is absent or represents -CH2-, -CH2CH2-, -CH2O- or -CH2NRd; Rd represents H or (1-4C alkyl); R3 represents H or (1-4C alkyl); and n is equal to 0-6. The radical values NRbRc, Y, hetAr1, hetAr2, hetAr3, hetCyc1, hetCyc2, NReRf, R4 are specified in the patent claim. The invention also refers to a pharmaceutical composition containing the above compounds, to a method of treating Trk kinase mediated diseases and conditions, such as pain, cancer, inflammation, neurodegenerative disease, Typanosoma cruzi infection, osteolytic disease, and to a method of preparing the above compounds.
EFFECT: invention refers to new derivatives of pyrazolo[1,5-a]pyrimidines possessing an inhibitory activity on tropomyosin-related kinases (Trk).
42 cl, 1 tbl, 105 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine and concerns anthracycline conjugates with carriers, such as antibodies; methods for preparing them; a pharmaceutical composition containing them; and to using them in treating tumours in mammals.
EFFECT: group of inventions provides the high and selective effect of tumour cell destruction.
36 cl, 12 ex, 32 dwg, 7 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine and concerns a method of treating meningioma in an individual in need thereof involving parenteral administration of a therapeutically effective amount of pasireotide or its pharmaceutically acceptable salt into the above individual; a compound for parenteral administration for treating meningioma containing pasireotide or its pharmaceutically acceptable salt together with one or more of their pharmaceutically acceptable solvents or carriers.
EFFECT: group of inventions provides better effectiveness and tolerance of pasireotide in patients with recurrent meningiomas than if applying the other treatment, eg with octreotide.
2 cl, 1 ex
SUBSTANCE: claimed is a pharmaceutical composition, which contains as minimum one active component - a pharmaceutically acceptable salt of 6-[(4-methyl-1-1-piperazinyl)methyl]-indolo[1',7':1,2,3]pyrrolo[3',4':6,7]azepino[4,5-b]indole-1,3(2H,10 H)-dione of formula (I), in particular mesylate of 6-[(4-methyl-1-1-piperazinyl)methyl]-indolo[1',7':1,2,3]pyrrolo[3',4':6,7]azepino[4,5-b]indole-1,3(2H,10 H)-dione of formula (1) and as minimum one auxiliary substance.
EFFECT: demonstrated suppression of growth of adenocarcinoma and melanoma tumour.
SUBSTANCE: invention represents a photosensitiser (PS) for photodynamic therapy (PDT) of cancer and other neoplasms of different genesis, as well as fluorescent diagnostics, where PS contains a trismeglumin salt of chlorine e6, and as a cryostabiliser - methyl glucamine in a ratio of 1:0.1-0.2 wt.p. The invention also relates to a method of obtaining the photosensitiser for PDT, which includes dilution of methylpheophorbide a in acetone, processing the obtained solution with aqueous alkali (NaOH or KOH) with following neutralisation of a reaction mixture by diluted hydrochloric acid to pH 4.5-5.0, separation of the precipitated sediment of chlorine e6 with following washing with distilled water, processing the sediment with a water solution and lyophilisation, which is characterised by the fact, that processing of methylpheophorbide a in acetone with aqueous alkali is carried out at a temperature of 40-50°C, separation of chlorine e6 sediment is realised by filtering through a layer of celite 545, and chlorine e6 is extracted from celite by a water solution of meglumine until the concentration of chlorine e6 salt, corresponding to optic density D=225-235/1 ml, is achieved.
EFFECT: obtaining a lyophilised drug form PS without ballast substances, which has an increased storage stability, which results in simplification and optimisation of chlorine e6 solution with a high degree of purity.
2 cl, 2 ex, 2 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine and can be applicable for the inhibition of tumour cell growth and proliferation. The tumour cell contacts with an extracellular matrix composition. The extracellular matrix composition is prepared by a human fibroblast cell culture on microcarrier granules in the hypoxic environment at the oxygen concentration of 1-5%.
EFFECT: group of inventions enables providing more effective action on the cancer cells.
24 cl, 18 dwg, 6 tbl, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to the pharmaceutical industry and concerns new derivatives of the antitumour antibiotic oligomycin A and a method for preparing them by regioselective [3+2]dipolar cycloconnection of an azido group of 33-deoxy-33-azidooligomycin A(1) to monosubstituted alkines. The new derivatives of the antibiotic oligomycin A described by formula: , wherein R represents 1,4-disubstituted 1,2,3-triazoles, namely a. - (phenyl-triazol-1-yl), b. - (4-carboxy-triazol-1-yl), c. - (4-4-methoxycarbonyl-triazol-1-yl), d. - (4-dimethylaminoethylamidocarboxytriazol-1-yl) possess manifested antitumour activity and higher solubility as compared to the initial oligomycin A.
EFFECT: preparing the new derivatives of the antitumour antibiotic.
3 cl, 4 dwg, 3 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to a pharmaceutical composition for the delivery of a pharmaceutical agent to a focus of a disease. The composition contains a water-insoluble pharmaceutical agent which is paclitaxel, a pharmaceutically acceptable carrier which is albumin, preferentially human serum albumin. The relation (wt/wt) of albumin to paclitaxel makes 9:1. The pharmaceutical composition contains nanoparticles containing paclitaxel and albumin wherein the nanoparticles have a size of less than 200 nm.
EFFECT: administering the pharmaceutical composition according to the invention provides enhanced characteristics of the delivery of paclitaxel to the site of the disease and reduced adverse side effects.
24 cl, 5 tbl, 51 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to a new derivative of uracil described by the following structural formula, and its pharmaceutically acceptable salt. The compound has the structural formula according to the R-enantiomer form: The given invention also refers to a method for preparing the above compound and to a pharmaceutical composition on the basis of the above compound. The method for preparing consists in a reaction of 5-fluoruracil and R-tert-butyl-3-iodopropane-1.2-diyldicarbamate.
EFFECT: compound possesses the properties of uridine phosphorylase inhibitors and can be used as an active ingredient for preparing a drug for treating cancer, such as an antimetabolite of the uridine phosphorylase inhibitors of tumour cells.
4 cl, 1 tbl, 1 ex
SUBSTANCE: invention represents a pharmacological geroprotective composition, which includes a polyphenol component, vitamins and microelements, humic acids, containing polyphenol components, vitamin C, vitamin A, iron (II) chloride and selenium (IV) dioxide, with the composition components being in a specified ratio in wt %.
EFFECT: increased life expectancy and retardation of tumour development.
2 cl, 3 dwg, 2 tbl, 4 ex