Improved method for synthesis of oxidised phospholipids

FIELD: chemistry.

SUBSTANCE: in the proposed methods, the hydroxyl group of the formula I compound is substituted with the corresponding residue so that use of column chromatography is avoided, , where A1 is CH2, R1 is alkyl, R3 is a residue of phosphoric acid, phosphorylcholine, phosphoryl ethanolamine, phosphorylserine, phosphoryl cardiolipin, phosphoryl inosite, ethylphosphocholine, phosphoryl methanol, phosphoryl ethanol, phosphoryl propanol, phosphoryl butanol, phosphoryl ethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propyleneglycol)], phosphoinosite-4-phosphate, phosphoinosite-4,5-biphosphonate, pyrophosphate, phosphoethanolaminediethylene-triaminepentaacetate, dinitrophenyl phosphoethanolamine, phosphogycerine or a reactive phosphorus-containing group which can convert to the said residues, or hydrogen.

EFFECT: design of methods of producing phospholipids, which avoid use of chromatography.

22 cl, 5 ex

 

The technical FIELD AND prior art TO WHICH the INVENTION RELATES.

The present invention relates to the field of synthetic chemistry and more particularly to a new synthetic methods applicable to obtain oxidized phospholipids and their derivatives, analogues and salts.

In the field of pharmacology modified phospholipids are known in many applications. In U.S. patent No. 5985292 described composition for percutaneous and transmembrane applications, including phospholipids, carrying soluble in lipids active compounds. In U.S. patent No. 6261597, 6017513 and 4614796 described phospholipid derivatives entered into liposomes, and Biofactory for delivering medicines. In U.S. patent No. 5660855 described made in liposomes lipid patterns aminoboronic derivatives of cholesterol that are suitable for targeting a cell or tissue of smooth muscle. These drugs are designed to reduce restenosis in arteries when applying the methods of RTSA.

The use of liposomes in the treatment of atherosclerosis, is also described in patent application PCT published as WO 95/23592. It describes pharmaceutical compositions adnyamathanha liposomes, which may contain phospholipids. Liposomes are described in WO 95/23592, are designed to optimize outflow of cholesterol from atherosclerotic plaques and are usually neonilin the mi phospholipids.

It is known that modified derivatives of phospholipid simulating patterns of platelet activating factor (PAF), are pharmaceutically active, affecting functions such as vascular permeability, blood pressure and reduced heart function. In U.S. patent No. 4778912 suggest that one group of such derivative possesses anticancer activity.

In U.S. patent No. 4329302 described synthetic compounds, a simple 1-On-alkalemia ester or 1-O-(fatty acyl)phosphoglyceride, which are derived lysolecithin applicable when mediating platelet activation. In U.S. patent No. 4329302 described that the acylation by acyl short circuit lysolecithin led to the formation of compounds with the ability to activate platelets in contrast to the compounds acylated by acyl long chain, and that a simple 1-On-alkilany ether biologically superior to the corresponding 1-O-(fatty acyl)derivatives in the simulation PAF.

The structural effect of different phospholipids on their biological activity was investigated Tokumura et.al. (Journal of Pharmacology and Experimental Therapeutics. July 1981, Vol.219, No.1) and in U.S. patent No. 4837011 in relation to their effect on hypertension.

In the patent Sweden CH 642665 described modified derivatives of ethers of fosfolipidov, which may have some physiological effect.

<> Davies et al. (J. Biol. Chem. 2001, 276:16015) describe the application of oxidized phospholipids as agonists of receptors activated proliferation peroxisome.

In U.S. patent No. 6838452 and in WO 04/106486 (which are hereby incorporated by reference in their entirety), which is the present assignee of this application, describes how to obtain well-characterized oxidized phospholipids, as well as other synthetic components of oxidized LDL (low density lipoprotein). Stated that the proposed compounds are highly effective in the treatment of atherosclerosis and related diseases, and autoimmune diseases and inflammatory disorders. The following describes that the oxidized phospholipid regulates the immune response to oxidized LDL. In addition, as described, which is usually converted into ethers of oxidized phospholipids are superior as therapeutic agents converted into esters of oxidized phospholipids.

Oxidation of phospholipids occurs in vivo through the action of free radicals and enzymatic reactions that are common in the atherosclerotic plaque. In vitro production of oxidized phospholipids usually includes a simple chemical oxidation of native LDL or phospholipid component of LDL. Researchers studying the role of oxidized LDL, used, for example, ions of iron(II) and ascorbic KIS the GTC (Itabe, H., et al., J.Biol.Chem. 1996; 271:33208-217) and copper sulfate (George, J. et al., Atherosclerosis. 1998; 138:147-152; Ameli, S. et al., Arteriosclerosis Thromb Vasc Biol 1996; 16:1074-79) to obtain oxidized or weakly oxidized phospholipid molecules, such molecules that are associated with components of the plaque. It is shown that in this way the resulting molecules are identical to the autoantigens associated with atherogenesis (A.D. Watson et al., J. Biol. Chem. 1997; 272:13597-607), and is able to induce protective anti-atherogenic immune tolerance in mice (Patent application U.S. No. 09/806400, Shoenfeld et al., registered on 30 September 1999). Similarly, in U.S. patent No. 5561052 describes a method for oxidized lipids and phospholipids using copper sulfate and superoxide dismutase to obtain oxidized arachidonic or linoleic acids and oxidized LDL diagnostic use.

The oxidation methods described above to obtain oxidized phospholipids include reactions that are nonspecific, and produce a mixture of oxidized products. Nespecificnomu reactions reduces output, requires additional stages of separation and increases the possibility of unwanted side effects, when the products are administered in pharmaceutical compositions.

1-Palmitoyl-2-(5-oxovalerate)-sn-glycero-3-phosphocholine (POVPC) and its derivatives, such as 1-Palmitoyl-2-glutaryl-sn-glice is about-3-phosphocholine (PGPC), are representative examples of weakly oxidized converted into esters of phospholipids, which are explored in relation to atherogenesis (see, for example, Boullier et al., J. Biol. Chem., 2000, 275:9163; Subbanagounder et al., Circulation Research, 1999, pp.311). Also studied the effect of various structural analogues, which belong to this class of oxidized phospholipids (see, for example, Subbanagounder et al., Arterioscler. Thromb. Nasc. Biol. 2000, pp.2248; Leitinger et al., Proc. Nat. Ac. Sci. 1999, 96:12010).

POVPC usually get a provision of phosphatidylcholine having unsaturated fatty acid, and oxidation of unsaturated communication fatty acids, for example by ozonolysis (oxidative cleavage) or using periodate as oxidizing agent. This synthetic path usually includes a multistage synthesis and requires separation of the majority of the formed intermediate product by column chromatography.

As described in U.S. patent No. 6838452, cited above, converted into ethers of oxidized phospholipids in this way was obtained by oxidation of unsaturated communication fatty acids attached to the main chain of the phospholipid. More specifically converted into ethers of oxidized phospholipids were obtained in accordance with the guidelines of this patent, the introduction of unsaturated fatty acids with short-chain in glycerolipid, the introduction of phosphate cha is t to obtain the intermediate product and the oxidation of unsaturated communication chain fatty acids using (i) hydrogen peroxide and formic acid so that to obtain the diol, followed by oxidation periodate potassium to obtain aldehyde; or (ii) ozonolysis. While the oxidative cleavage of unsaturated communication leads to the formation of aldehyde groups, other oxidized groups (e.g. carboxylic acid, acetal, etc.) received further oxidation of the aldehyde group. Such multi-stage synthetic path is often characterized by relatively low overall yields and again requires separation of the majority of the formed intermediates column chromatography.

It was found that the application of in vivo converted into esters of oxidized phospholipids obtained as described above have the disadvantage of sensitivity related to the recognition, binding and metabolism of the active ingredient in the body, giving importance to the dosing and stability after injection. Converted into ethers, oxidized phospholipids, such as described in U.S. patent No. 6838452 and in WO 04/106486, exhibit higher biostability and high therapeutic activity.

Thus, presently known methods for producing transformed into the ethers and esters of oxidized phospholipids include complex multi-stage techniques suitable for laboratory receipt, however, play get in about Islena scale is inefficient and complex. In particular, these multistage methods require unacceptable in the industry such separation techniques as column chromatography, at various stages of the synthesis.

Due to the useful therapeutic activity of oxidized phospholipids, in General, and converted into ethers of oxidized phospholipids, in particular, there is a widely recognized need for an improved method of obtaining converted into ethers of oxidized phospholipids, devoid of at least some of the disadvantages of the methods known in the art, and this improved method may be very appropriate.

The INVENTION

According to one aspect of the present invention, a method for obtaining compounds with glycerin main chain and at least one containing oxidized group, residue, attached to glycerol main chain through an ether linkage, which includes providing the first connection with glycerin main chain and at least one free hydroxyl group; providing a second compound having at least one unsaturated bond and at least one reactive group capable to form an ether linkage with a free hydroxyl group; the interaction of the first connection and the second connection is the means to obtain thereby a third connection, and the third compound is glycerol main chain and containing unsaturated bond residue attached to glycerol main chain through an ether linkage; the allocation of a third connection to receive it in a purified form; the interaction of the purified third compound with an oxidizing agent to obtain thereby the fourth connection and the fourth connection has glycerin main chain and containing oxidized group residue attached to glycerol main chain through an ether linkage; and the allocation of the fourth connection for receiving the thus purified fourth compound, thus obtaining the compound with glycerin main chain and at least one containing oxidized group residue attached to glycerol main chain through an ether bond in a way that avoids column chromatography.

In accordance with the following characteristic features in preferred embodiments of the invention described below, the interaction of the first connection and the second connection is carried out in the presence of a base.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of a base selected from the group consisting of sodium hydroxide, lithium hydroxide, hydroxide feces is tion, of barium hydroxide and potassium hydroxide.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the reactive group is a halide.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the allocation of a third connection involves collecting a third connection; obtaining the solution of the third compound in the solvent, whereby the solvent is chosen so that the third compound was soluble in it, and impurities formed during the reaction, were insoluble therein, to thereby obtain a mixture comprising a solution of a third compound in the solvent and insoluble impurities; removing insoluble impurities; and removing the solvent, thereby obtaining a purified third compound.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of a solvent selected from the group consisting of petroleum ether, hexane and benzene.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of an oxidizing agent selected from the group consisting of formic acid, hydrogen peroxide, periodate, perchlorate, bismuthate, permanganate, chlorite, ozone, silver oxide, osmium tetroxide, and any combination thereof.

With the according to the following characteristic features in the described preferred embodiments, the implementation of the oxidized group selected from the group consisting of carboxylic acids, complex, ester, aldehyde, acetal, Catala and diol.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the oxidized group is an aldehyde and the interaction of the purified third compound with an oxidizing agent includes the transformation of the purified third compound in connection with glycerin main chain and volaterrae residue attached in glycerol main chain through an ether linkage; and oxidation of compounds having glycerin main chain and volaterrae residue attached to glycerol main chain, to get through the fourth connection with glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the transformation carried out by the interaction of the purified third compound with the first oxidizing agent selected from the group consisting of peroxide, bismuthate, periodate, permanganate, and any combination thereof.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the oxidation is performed by the interaction of compounds with glycerin main chain and volaterrae residue, attached to glycerol main chain, with a second oxidizing agent selected from the group consisting of periodate, bismuthate, permanganate and chlorite.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the allocation of the fourth connection includes: collecting the fourth connection; obtaining water-soluble adduct of the fourth connection; the transformation of the water-soluble adduct in a two-phase system to get through this aqueous phase containing adduct, and the organic phase containing the water-insoluble impurities formed during the interaction with an oxidizing agent; collecting the aqueous phase; the decomposition of the adduct; and collecting the fourth connection to receive thereby cleaned fourth connection.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the receiving water-soluble adduct involves interaction of the fourth connection reagent Gerard.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the oxidized group is a carboxylic acid, and the interaction of the purified third compound with an oxidizing agent and includes the transformation of the purified third compound in connection with glycerine the th main chain and aldehydebase residue, attached to glycerol main chain through an ether linkage; and oxidation of compounds having glycerin main chain and aldehydebase residue attached to glycerol main chain, to obtain thereby the connection with glycerin main chain and containing carboxylic acid residue attached to the glycerol main chain through an ether linkage.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the transformation of the purified third compound in connection with glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage, includes the transformation of the purified third compound in connection with glycerin main chain and volaterrae residue attached to glycerol main chain through an ether linkage; and oxidation of compounds having glycerin main chain and volaterrae residue attached to glycerol main chain, to obtain thereby the connection with glycerin main chain and aldehydebase residue attached to glycerol main chain through simple radio link.

In accordance with the following characteristic features in the described preferred option is the implementation of the method further includes allocating a connection, with glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage, to obtain thereby a purified compound having glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the allocation involves collecting connection with glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage; obtaining water-soluble adduct compounds having glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage, as described above; the transformation of the water-soluble adduct in a two-phase system to obtain thereby the aqueous phase containing the complex, and the organic phase containing the water-insoluble impurities formed during the conversion and/or oxidation; collecting water phase; the decomposition of the adduct; and collecting connection with glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage, to obtain thereby a purified connect the tion, with glycerin main chain and aldehydebase residue attached to glycerol main chain through an ether linkage.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the oxidized group is a carboxylic acid, and the interaction of the purified third compound with an oxidizing agent includes the transformation of the purified third compound in connection with glycerin main chain and epoxydodecane residue attached to glycerol main chain through an ether linkage; and oxidation of compounds having glycerin main chain and epoxydodecane residue attached to glycerol main chain, to obtain thereby the connection with glycerin main chain and containing carboxylic acid residue attached to the glycerol main chain through an ether linkage.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the transformation involves the interaction of the third connection peroxide.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the first connection has at least two free hydroxyl groups, the method further includes the about the interaction between the first connection and the second connection protection at least one of the at least two groups protecting group.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the protective group is trityl.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the first connection has at least two free hydroxyl groups, the method further includes prior to the engagement of the first connection and the second connection protection at least one of the at least two groups protecting group, preferably trailvoy group.

In accordance with the following characteristic features in the described preferred embodiments, implementation, when the method includes education epoxydodecane connection, as described above, the method further includes prior to the engagement of the third compound and oxidizing agent replacement trityl protecting group selected from the group consisting of acetate, pivaloate or benzoate.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the connection with glycerin main chain and at least one containing oxidized group, residue, attached to glycerol main chain, optionally includes a phosphate part attached to glycerol main chain, and the method further in the cancel up to the interaction between the first connection and the second connection to highlight the third connection to the interaction of the third compound with an oxidizing agent, prior to the allocation of the fourth connection or after allocation of the fourth connection:

the interaction of the first connection, the third connection, the purified third compound, a fourth compound or purified fourth compound with fosforsoderzhashchie part to obtain thereby a connection having glycerin main chain and at least one containing oxidized group residue attached to glycerol main chain and optionally having a phosphorus-containing portion attached to glycerol main chain.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of at least one phosphorus-containing component is a phosphate group that is attached to a glycerol main chain through fosfodiesterazu connection.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of at least one part comprising the phosphorus-containing group selected from the group consisting of phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphoryl tanolin-N-lactose, the phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine and fosfoglitserin.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the phosphorus-containing portion is attached to the sn-3 position of glycerol main circuit connection.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the interaction of the first connection, the third connection, the purified third compound, a fourth compound or cleaned fourth connection part which includes a phosphorus-containing group, includes obtaining a first connection, the third connection, the purified third compound, a fourth compound or purified fourth compound with a free hydroxyl group; the interaction of the first connection, the third connection, the purified third compound, a fourth compound or purified fourth compound with a reactive compound having a phosphorus-containing group with the second reactive group and a third reactive group, the second reactive group capable of interacting with the free hydroxyl groups of which the first and the second reactive group to obtain thereby the first connection, the third connection, the purified third compound, a fourth compound or purified fourth compound having a group reactive phosphorous-containing group attached to the glycerol main chain; and the transformation of the group, reactive fosforsoderzhashchie group in phosphorus-containing part.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the compound with a reactive phosphorous-containing group, a represents a phosphorous oxychloride (POCl3).

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the interaction is carried out in the presence of a base.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the part having the phosphorus-containing group is a phosphoric acid, and the transformation involves the hydrolysis part comprising a reactive phosphorous-containing group.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the part having fosforsoderzhashchih group includes aminoalkyl group, and the transformation involves interaction group having phosphorus-containing group derived aminoalkyl group, and the PR is izvozna capable of interacting with a third reactive group.

According to another aspect of the present invention is proposed another method of obtaining compounds with glycerin main chain and at least one residue containing oxidized group attached to the glycerol main chain through an ether linkage, and the method includes obtaining a first connection with glycerin main chain and at least one free hydroxyl group; obtaining a fifth connection having at least one oxidized group and at least one fourth reactive group; the interaction of the first connection and the fifth connection obtained through this reaction mixture, containing the sixth connection, and the sixth connection has glycerin the main chain and at least one residue containing oxidized group attached to the glycerol main chain through an ether linkage; and allocating a connection with glycerin main chain and at least one residue containing oxidized group attached to the glycerol main chain through an ether linkage.

According to the following characteristics in preferred embodiments of the invention described below, the interaction of the first connection and the fifth connection is performed in the presence of a base.

With the according to the following characteristic features in the described preferred embodiments, the implementation of a base selected from the group consisting of sodium hydride, sociallyengaged, sodium amide, sodium hydroxide and any mixture thereof.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the fourth reactive group is a halide.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the oxidized group selected from the group consisting of carboxylic acids, complex ether, allhelgona, aldehyde, acetal, Catala and diol.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the fifth compound contains less than 4 carbon atoms.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the fifth compound contains more than 5 carbon atoms.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the fifth connection has at least two free hydroxyl groups, and the method further includes prior to the engagement of the first connection and the fifth connection protection at least one of the at least two groups protecting group.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the protective group is trityl.

Coz the ACLs the following characteristic features in the described preferred embodiments, the implementation of the connection, with glycerin main chain and at least one containing oxidized group residue attached to glycerol main circuit additionally includes having a phosphate group part attached to glycerol main chain, and the method further includes, before or after the interaction of the first connection and the fifth connection, or after separation of the sixth connection the interaction of the first connection or the sixth connection part which includes a phosphate group, thereby obtaining compounds having glycerin main chain and at least one containing oxidized group balance, prisoedinenii to glycerol main chain and optionally having a phosphorus-containing part attached to glycerol main chain as described above.

In accordance with the following characteristic features in preferred embodiments of the invention, described below, in any of the ways described here, the first connection further comprises at least one alkylenes chain having 1-30 carbon atoms.

In accordance with the following characteristic features in the described preferred embodiments, the implementation Allenova chain attached to glycerol main chain through an ether linkage.

In accordance with the following characteristic p is iznakam in the described preferred embodiments, the implementation Allenova chain attached at the sn-1-position of glycerol main chain of the first connection.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the residue containing the oxidized group attached at the sn-2-position of the compound and, in addition, where at least one of the at least one free hydroxyl group of glycerol main chain is in the sn-2-position of the first connection.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the first compound has the General formula I:

The formula I

in which:

A is absent or selected from the group consisting of CH2, CH=CH and C=O;

R1selected from the group consisting of H and hydrocarbon chain having 1-30 carbon atoms; and

R3selected from the group consisting of hydrogen, alkyl, aryl, phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrophenolate lamina, fosfoglitserin.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the connection with glycerol portion and at least one oxidized group attached to the glycerol main chain through an ether linkage, has a General formula II:

Formula II

in which:

And1selected from the group consisting of CH2, CH=CH and C=O;

And2represents CH2;

R1represents alkyl having 1-30 carbon atoms;

R2represents a

where X is an alkyl chain having 1-24 carbon atoms;

Y is selected from the group consisting of hydrogen, hydroxy, alkyl, alkoxy, halogen, acetoxy and aromatic functional groups; and

Z is selected from the group consisting of:

and R4is alkyl or aryl; and

R3selected from the group consisting of hydrogen, alkyl, aryl, phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, FOSFA relationally-N-lactose, the phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine, fosfoglitserin.

According to another aspect of the present invention, a method of introducing a phosphate part in connection with glycerin main chain and having a residue containing oxidized group or pre-oxidized group attached thereto through an ether linkage, which includes obtaining compounds having glycerin main chain and the residue containing the oxidized group or pre-oxidized group attached to the glycerol main chain through an ether linkage, and at least one free hydroxyl group; the interaction of compounds, including phosphorus-containing group with the second reactive group and a third reactive group, the second reactive group capable of interacting with a free hydroxyl group thereby obtaining compounds having a residue containing oxidized group or pre-oxidized group, and a reactive group comprising phosphorus-containing group; and the transformation of the reactive group comprising phosphorus-containing group, FOS is atnow part, introducing thereby the phosphate part in the connection.

In accordance with the following characteristic features in preferred embodiments of the invention described below, the connection with glycerin main chain comprises at least one alkylenes chain having 1-30 carbon atoms.

In accordance with the following characteristic features in the described preferred embodiments, the implementation Allenova chain attached to glycerol main chain through an ether linkage.

In accordance with the following characteristic features in the described preferred embodiments, the implementation Allenova chain attached at the sn-1-position of glycerol main circuit connection.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the oxidized group selected from the group consisting of carboxylic acids, complex ether, allhelgona, aldehyde, acetal, diol and Catala.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of pre-oxidized group represents an unsaturated group.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the connection involving the phosphate group is POCl3.

In accordance with the following characteristic features in sannich preferred embodiments, the implementation of the interaction produced in the presence of a base.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the base is a tertiary amine.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of a connection having a phosphate group is POCl3and a group containing a reactive phosphorous-containing group is dichlorophosphate group.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the connection with glycerin main chain has a residue containing the pre-oxidized group attached thereto through an ether linkage.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of a phosphate selected from the group consisting of phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, phosphorylethanolamine, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine, fosfoglitserin.

In accordance with the following characteristic is sternum features in the described preferred embodiments, the implementation of the phosphate component is phosphoric acid, and the transformation involves the hydrolysis part comprising a reactive phosphorous-containing group.

In accordance with the following characteristic features in the described preferred embodiments, the implementation of the phosphate part contains alkylamino, and the transformation involves interaction part comprising a reactive phosphorous-containing group, with a derivative of aminoalkyl, and derivative capable of interacting with a part that includes a reactive phosphorous-containing group.

The present invention successfully overcomes the shortcomings of the presently known methods offer new synthetic routes that can be advantageous to use when getting in an enlarged scale of oxidized phospholipids.

Unless otherwise noted, all technical and scientific terms used herein have the same meaning as commonly understood by any average expert in the field to which this invention relates. Although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present invention, suitable methods and materials are described below. In case of conflict, the description of the invention, including definitions, will control. In addition, substances, methods and examples are illustrative only and are not intended the La limit the scope of the claims.

Used here, the term "mixture" refers to a mixture that includes more than one substance and which may be in any form, for example as a homogeneous solution, suspension, dispersion, two-phase solution, and so forth.

Used in this application, the singular "a", "an" and "the" include the plural, unless the context clearly indicates otherwise.

Throughout this description, various aspects of this invention can be presented in a range format. It should be clear that the description in range format is merely for convenience and brevity, and will not be construed as a strict limitation on the scope of the invention. Accordingly, it should be considered that the description of the interval is specifically described all possible potentially, as well as individual numerical values within an interval. For example, consider that the description of such interval, as 1-6, specifically describes such potentially as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc., as well as individual numbers within an interval, such as 1, 2, 3, 4, 5 and 6. This description is used regardless of the width of the interval.

Have in mind that every time there specified numerical interval includes any cited the number (single or combined) in the specified interval. The phrase "located/is situated between the first specified number, and the second is shown by the number and average/ranges from" a first specified number of "before" the second specified number used here interchangeably and mean, they include the first and second indicated numbers and all the individual and combined numbers between them.

Used here throughout, the terms "includes", "including" and "comprising" means that there may be added other stages and ingredients that do not affect the final resultatet terms include the terms "consisting of" and "consisting essentially of".

The phrase "consisting essentially of" means that the composition or method may include additional ingredients and/or the stage, but only if the additional ingredients and/or phase of the material do not change the basis and novel characteristics of the claimed composition or method.

The term "method" or "method" refers to the methods, tools, techniques and methods to perform this task, including, but not limited to those listed methods, tools, techniques and methods, either known to, or readily developed from known methods, tools, techniques and methods practitioners in the chemical, pharmaceutical, biological, biochemical and medical fields.

The term "phospholipid" is used here to a General description of compounds, which include non-polar lipid group, and a highly polar terminal phosphate group. One particular and the most dominant in the nature of families, the creation of phospholipid compounds is a family of compounds of phosphoglycerides. The term "phospholipid" is therefore usually applied throughout to describe phosphoglycerides, if not specified.

The term "phosphoglyceric" therefore, is used here to describe compounds with glycerin main chain, one or more lipid parts and one or more phosphate terminal group are attached to glycerol main chain. Most of glycerolipids of natural origin include two lipid part attached at the sn-1 and sn-2 positions, and one phosphate part attached to the sn-3 position of glycerol main chain.

The term "oxidized phospholipid" is therefore used here to describe the phospholipid and phosphoglyceride, which includes one or more oxidized groups, as that term is described below. Typically, the oxidized oxidized phospholipids group included in the lipid part.

The term "glycerolipid" describes a compound that glycerin main chain and one or two lipid part attached to it. The lipid part can be attached to glycerol main chain via ester or ether linkage.

Used here, the term "lipid" refers to a hydrocarbon residue having 3-30 carbon atoms. In compounds of natural origin lipids in the phospholipid and glycerolipid formed from fatty KIS is from and so attached to the main chain through a linkage O-acyl (ester bond). Here the lipid portion can be attached to the main chain or through an ether or ester bond.

Used here, the terms "mono-esterified and di-esterified" against phospholipids or glycerolipids describe phospholipids or glycerolipid either oxidized or unoxidized, in which one or two of lipid parts respectively attached to the glycerol main chain via ester (for example, O-fatty acyl) connection.

Used here, the terms "simple monoamine" and "simple diepiriye derived in respect of phospholipids and glycerolipids describe phospholipids or glycerolipid either oxidized or unoxidized, in which one or two lipid parts respectively attached to the glycerol main chain through an ether linkage.

The term "fosfoglitserin" describes a compound that glycerin main chain and a phosphate group attached to one of the provisions of the chain.

The term "phosphoglyceride" describes a compound that glycerin main chain, one or two lipid part and attached thereto phosphate part.

The term "having ether group phosphoglyceric" describes phosphoglyceride, in which the lipid portion attached to glycerol main chain through an ether linkage.

Used here is Ermin "part describes the functional substance or group, which forms part of the connection.

The term "residue", as is well known in this field, apply here described for the main part of the molecule, which is associated with another molecule.

DESCRIPTION of the PREFERRED embodiments

The present invention offers new ways of producing oxidized phospholipids, which can be effectively used to obtain an enlarged scale of these oxidized phospholipids. In particular, the present invention provides new ways of introduction of oxidized groups in connection with glycerin main chain, and also offers new ways of introducing part comprising phosphorus-containing group, in this connection. New ways described here, avoid using column chromatography and for them is usually used commercially available and environmentally friendly reagents.

The principles and operation of new synthetic methods according to this invention can be better understood with reference to the accompanying description.

Before Abyssinian at least one variant embodiment of the invention in detail, it should be clear that the invention is not limited in its application to the details set forth in the following description or illustrated by examples. The invention allows for other ways to implement or may be is to be undertaken or carried out in various ways. Should also be understood that the phraseology and terminology used here, is intended for the purpose of description and should not be construed as limiting.

As discussed above, recently described that well characterized, obtained by synthetic oxidized phospholipids can regulate the immune response to oxidized LDL and are therefore highly effective in the treatment of atherosclerosis and related diseases, and autoimmune diseases and inflammatory disorders. In addition, as described, which is usually converted into ethers of oxidized phospholipids as therapeutic agents are superior compared to converted into esters of oxidized phospholipids.

These are very useful oxidized phospholipids typically include glycerin main chain, to which the lipid residue, a phosphate residue and the lipid residue containing oxidized group attached, as described, for example, in U.S. patent No. 6838452 and in WO 04/106486.

As further discussed above, currently known methods for producing such well-characterized synthetic oxidized phospholipids include multi-stage syntheses. It was found that while these multistage syntheses are relatively effective, giving in d is the query result in average good output these methods are limited by neobhodimosti to produce the isolation and purification of laboratory methods of various intermediate products formed during the synthesis. In particular, these methods usually include techniques such as column chromatography, which, as is widely recognized by experts in this field, is industrially unacceptable, or at least inefficient in terms of cost, complexity and use of excessive quantities of organic solvents, which can be dangerous and require special care when removing waste. The necessity of using column chromatography in these methods derives from the fact that intermediate products, and final products formed during these multistage syntheses, it is impossible to remove and/or clean other more satisfying technical conditions by techniques such as extraction, crystallization and the like.

Because those obtained by synthetic oxidized phospholipids demonstrate extremely useful therapeutic activity, it is highly desirable to obtain these compounds with a higher level of purity. In addition, since the receipt of such oxidized phospholipids includes multistage syntheses requires purification of intermediate products, in order to implement the giving of such method gave the appropriate outputs, with minimal by-products.

In the search for enhanced ways of getting oxidized phospholipids, which can be effectively used in the production in an enlarged scale of these compounds, at the same time avoiding the necessity of applying laboratory techniques, such as column chromatography, the authors present invention has developed and successfully implemented in practice advanced synthetic methodologies for the introduction of oxidized groups and/or phosphate part in compounds that have glycerin main chain, which excluded the adverse application of column chromatography and which lead to relatively high outputs pure compounds. In the methods described here, in addition, is usually used commercially available, non-hazardous reagents, which, in addition, provide an opportunity for their industrial application.

New synthetic methodology described here can be divided as the following:

(i) a new way of introducing the oxidized groups in connection with glycerin main chain, by introducing an unsaturated group and oxidation of unsaturated groups, resulting in an after-mentioned oxidation compound containing oxidized group, is isolated and purified by a water-soluble adduct;

(ii) new is th way of the introduction of oxidized groups in connection with glycerin main chain, by introducing an unsaturated group and oxidation of unsaturated groups, resulting in the said oxidation is performed via epoxy intermediate product in the presence of selective protective group;

(iii) a new way of introducing the oxidized groups in connection with glycerin main chain, through direct injection of oxidized groups; and

(iv) an improved method of introducing a phosphate part in glycerolipid, with optional attached or pre-oxidized oxidized group, through the introduction of the reaction fosforsoderzhashchie group.

Thanks to the superior efficiency of oxidized phospholipids in which the residue containing oxidized group attached to the main chain through an ether linkage, all these methods are aimed at the accession of residue containing oxidized group, glycerin main chain through an ether linkage.

As shown in the examples section which follows from the application of these methodologies are well characterized by oxidized phospholipids successfully received with a relatively high yield and purity.

Thus, according to one aspect of the present invention, a method for obtaining compounds with glycerin main chain and at least the Dean balance, containing oxidized group attached to the glycerol main chain through an ether linkage, which eliminates the use of column chromatography. The method according to this aspect of the present invention is carried out by:

obtaining a first connection with glycerin main chain and at least one free hydroxyl group;

obtain a second compound having at least one unsaturated bond and at least one reactive group capable of forming simple radio communication with the said free hydroxyl group;

the interaction of the first connection and the second connection with obtaining thereby a third connection, which has glycerin main chain and residue containing unsaturated bond, which are attached to glycerol main chain through an ether linkage;

the allocation of a third connection with obtaining thereby a purified third compound;

the interaction of purified third compound with an oxidizing agent thereby obtaining a fourth connection, which has glycerin main chain and containing oxidized group residue attached to glycerol main chain through an ether linkage; and

the allocation of the fourth connection with obtaining thereby cleaned fourth the CSOs connection getting through this connection with glycerin main chain and at least one containing oxidized group residue attached to glycerol main chain through an ether linkage.

Used here throughout, the phrase "in connection with glycerin main chain", which is also called here interchangeably "glycerin connection" or "glycerin compound"describes a compound that includes the following structure:

When the compound is glycerol, each of positions of the glycerol sn-1, sn-2 and sn-3 is substituted by a free hydroxyl group.

Used here throughout phrase "oxidized group and a residue containing oxidized group", which are used here interchangeably, describe the organic part, in which at least one of its carbon atoms replaced by oxygen atom. Examples, without limitation, include aldehyde, carboxylic acid, ester of carboxylic acid, diol, acetal and ketal. The phrase "a compound having a residue containing oxidized group and the compound containing oxidized group" is also used here interchangeably.

The method according to the first aspect of the present invention is based on the introduction of unsaturated groups in glycerol of the giving and implementation of the oxidative cleavage of unsaturated linkages. However, because this synthetic path was used in currently known syntheses glycerin oxidized phospholipids, the authors of the present invention in this paper have been developed and successfully carried out almost this method, in which the glycerin compound, which has attached to it the oxidized group, you can select and clear without the use of column chromatography.

Introduction unsaturated groups in glycerol compound is usually carried out using known in the field of methods, such as described, for example, in U.S. patent No. 6838452.

Usually the first connection, which has glycerin main chain and at least one free hydroxyl group, is chosen as starting material.

A compound that has an unsaturated group and a first reactive group, which is also designated here as a second connection, receive any commercial manner or using methods known in this field, and subjected to reaction with glycerin original substance.

Choose the first reactive group capable of interacting with a free hydroxyl group. The interaction with the free hydroxyl group to form ether bond, usually performed via nucleophilic mechanism, and therefore is of erva reactive group is preferably characterized as a good leaving group, it may be, for example, a halide, sulfonate and another leaving group.

Preferably reactional group is a halide and more preferably a bromide.

A second connection is preferably chosen so that the unsaturated group is present in its end portion so as to facilitate the following oxidation reaction. "Unsaturated group" here means a group that includes at least two carbon atoms, which are connected between an unsaturated bond such as a double bond or triple bond, preferably a double bond.

Also preferably, the second compound contains 4-30 carbon atoms, more preferably from 4 to 27 carbon atoms, more preferably 4-16 carbon atoms, more preferably 4-10 carbon atoms, more preferably 4 to 8 carbon atoms and most preferably the second compound contains 6 carbon atoms.

The interaction of the first connection and the second connection described here, usually carried out in the presence of a base. Suitable bases for use in this context of the present invention include, without limitation, inorganic bases such as sodium hydroxide, lithium hydroxide, calcium hydroxide, barium hydroxide and potassium hydroxide.

The interaction of the first connection and the second connection Oba is but carried out in the presence of a solvent. Suitable solvents for use in this context of the present invention include, without limitation, non-polar solvents such as petroleum ether, hexane, benzene and toluene.

In cases when it is required to carry out the reaction selectively, namely the introduction of unsaturated groups in a certain position glycerin main chain, a free hydroxyl group, other than reactive hydroxyl, if present, must be protected prior to the interaction.

Thus, in such cases, the method according to this aspect of the present invention optionally and preferably additionally includes communication with the first connection and the second connection protection for one or more free hydroxyl groups which may be present in the first connection.

Any of the known hydroxyamine groups can be used in this context of the present invention. According to a preferred variant implementation of this aspect of the present invention, the protective group is trityl.

Trail is a three-dimensional group, which is usually used as a selective protective group due to spatial problems. Thus, despite the interaction of glycerol compound that has more than one free Hydra is xinnuo group, usually triticina group will react with less hindered by the group.

As noted above and, in addition, discussed in detail in U.S. patent No. 6838452 and in WO 04/106486, the position of the glycerol main chain, which is attached oxidized group influences the activity of the compounds. Therefore, it is very advantageous to make glycerin compounds described herein, selectively, so that the residue containing oxidized group, was attached in the desired position. As further indicated in U.S. patent No. 6838452, oxidized phospholipids, which have a residue containing oxidized group attached at the sn-2 position of the glycerol main chain, have a higher efficiency.

For such purpose applying trailvoy group as a protective group in the introduction above, the second connection in glycerol main chain is very successful due to its volume, protection of hydroxyl end groups in the sn-1 and/or sn-3 position to be effective, leaving a hydroxyl group at the sn-2-position available for substitution. After the interaction between the first connection and the second connection is completed, receive a reactive mixture which contains a compound which has glycerin main chain and residue containing unsaturated group attached to it h the rez simple radio communication. This connection also indicate here interchangeably as a third connection.

Depending on the source of the substance the third compound may further include a protective group, protecting the free hydroxyl groups which may be present in glycerol main chain.

The third connection is either protected or unprotected then separated from the reaction mixture and process to obtain purified compound.

In a preferred embodiment, the selection of the third connection is carried out primarily by the collection formed the third connection. Collection of the third connection is usually performed with the use of such conventional techniques, such as extraction, removal of solvent, filtration and the like, including any combination of them. After collecting the crude product is dissolved in a solvent, whereby the solvent is chosen such, in which the third compound is dissolved, while the impurities formed during the reaction between the first and second connections, it does not dissolve.

The term "impurities" is used here to describe any substance that is present in the final crude product itself is not a product, and includes, for example, unreacted educt and by-products.

With application is m such solvent receive the mixture, which includes the solution of the third compound in this solvent and insoluble substances. Suitable solvents for use in this context of the present invention include, without limitation, petroleum ether, hexane, benzene, heptane and toluene. Preferably the solvent is petroleum ether.

Insoluble impurities are then removed from the mixture, preferably by filtration, the solvent is removed and get cleaned third connection that eliminates the need for column chromatography in a method of cleaning it.

The purified third compound then reacts with an oxidizing agent to oxidize unsaturated group and thereby to obtain a fourth connection, in which the residue containing oxidized group attached to the glycerol main chain through an ether linkage.

The oxidizing agent is chosen depending on the desired oxidizable groups, as described in detail here below, and may be, for example, peroxide, periodate, visatec, permanganate, chlorite, ozone, silver oxide, osmium tetroxide and any combination of them.

Used here, the term "periodic" describes a compound having the formula XIO4where X can be hydrogen (for period acid) or monovalent metal cation (e.g. sodium, potassium).

The term "in smutt" describes the connection having the formula XBiO3where X can be hydrogen or monovalent metal cation (e.g. sodium, potassium).

The term "permanganate" describes a compound having the formula XMnO4where X can be hydrogen or monovalent metal cation (e.g. sodium, potassium).

The term "chlorite" describes a compound having the formula XClO2where X can be hydrogen or monovalent metal cation (e.g. sodium, potassium). Used here, the term "peroxide" includes a compound having the formula R-O-O-H, where R can be hydrogen, alkyl, cycloalkyl, aryl, oxyalkyl, oxocyclohexyl and oxiella, as these terms are define here.

Used here throughout, the term "alkyl" refers to saturated aliphatic hydrocarbon, including unbranched chains and branched chains. Preferably the alkyl group has 1-20 carbon atoms.

Group "cycloalkyl" refers to an all-carbon monocyclic or condensed cyclic groups (for example cycles which share adjacent pairs of carbon atoms)in which one or more of the cycles does not have a completely conjugated PI-electron system. Examples cycloalkyl groups, without limitation, are cyclopropane, CYCLOBUTANE, cyclopentane, cyclopentene, cyclohexane, qi is logication, Cycloheptane, cycloheptatrien and adamantane.

Group "aryl" refers to uglerodnym monocyclic or condensed polycyclic cyclic groups (for example, cycles which share adjacent pairs of carbon atoms)having a fully conjugate PI-electron system. Examples of aryl groups, without limitation, are phenyl, naphthalenyl and anthracene.

The terms "oxyalkyl", "oxocyclohexyl" and oxyaryl describe R'-C(=O)-group in which R' represents alkyl, cycloalkyl or aryl, respectively, so that the peroxide is peroxycarbonates acid.

Preferably the peroxide is hydrogen peroxide or peroxocarbonate acid.

Thus, in one embodiment of this aspect of the present invention group is oxidized to the aldehyde, and the interaction of the third compound with an oxidizing agent is carried out first by turning unsaturated group in the third connection diol group, preferably with the aid of an oxidizing agent, which is called here as the first oxidizing agent, and then further oxidation of diol groups by means of the second oxidizing agent in the aldehyde group.

The first and second oxidizing agents may be the same or different and can be, for example, peroxide, periodate, vimutta, what permanganate, chlorite, ozone, and any combination thereof.

In cases where the first and second oxidizing agents are the same, and depending on the oxidizing agent for the conversion of unsaturated groups in the diol group and oxidation of diol groups can be run simultaneously. Suitable oxidizing agents which can be used in this regard include oxidizing agents, which are able to induce oxidative cleavage of unsaturated groups such as, for example, ozone, osmium tetroxide and potassium permanganate.

In cases where the first and second oxidizing agents are different, the first preferred oxidizing agent is a peroxide, such as hydrogen peroxide, and the second oxidizing agent is, for example, periodic or wismuth.

Reaction conditions under which implement the methods of transforming and oxidation, determined in accordance with the applicable oxidizing agent.

In the preferred embodiment of this aspect of the present invention the first and second oxidizing agents are different, and the transformation of unsaturated groups in the diol group and oxidation of the diol group exercise consistently. In addition, according to a preferred variant implementation of this aspect of the present invention, after received diol, the protective group, if presets is there, removed to obtain the compound having three or more free hydroxyl group (here, Tirol). This connection can easily be cleaned prior to its oxidation to the aldehyde by means of crystallization, due to its unique chemical characteristics, as shown in the examples section hereinafter (see example 1). After cleaning, before the next stage of the synthesis is possible to perform selective protection of the free hydroxyl group at the sn-1 and/or sn-3 position.

Thus formed aldehydebase glycerol compound is then isolated from the reaction mixture and purified.

In the preferred embodiment of this aspect of the present invention, the aldehyde is purified by means of formation of the water-soluble adduct.

Thus, after the interaction with the oxidizing agent (agents) completes aldehydebase fourth connection harvested using conventional methods, as described herein above, and then the crude product is converted into its water-soluble addactionlistener such transformations in a two-phase system, receive the aqueous phase which contains a water-soluble adduct, and the organic phase, which contains water-insoluble impurities. Since most of the side products and unreacted substances, forming the s during the oxidation reaction, are organic substances, such substances are easily separated from water-soluble adduct collecting the aqueous phase. After that aldehydebase connection is removed by decomposition of water-soluble adduct.

Suitable water-soluble adducts, which can be applied in this context, the present invention is preferably produced by interaction aldehydebase connection with the reagent Gerard.

Reagents Gerard are a family of substances which are capable of forming water-soluble gidrazonami adducts with carbonyl-containing compounds, and thus allows the separation of carbonyl-containing compounds from other decarbonising compounds. Reagents Gerard represent ionic derivatives of semicarbazide.

The t-shape is a hydrazide chloride (carboxymethyl)trimethylammonium:

D-form is a hydrazide chloride (carboxymethyl)dimethylammonio:

Form R is a hydrazide chloride 1-(carboxymethyl)pyridinium:

Thus, the transformation aldehydebase compounds in water-soluble adduct with the reagent Gerard easy and convenient to get cleaned fourth connection, at the same time to exclude the application of column chromium is ographie.

When oxidized group is a carboxylic acid, the interaction of the third compound with an oxidizing agent can be performed first obtaining aldehydebase compounds, optionally and preferably as described herein above, and also, optionally and preferably the processed aldehydebase connection with the use of the methodology described here above, and then further oxidation of aldehyde to carboxylic acid.

Oxidation of aldehyde to carboxylic acid is preferably carried out by the interaction of the aldehyde with an oxidizing agent such as chlorite.

Alternatively, the unsaturated group can oxidize to carboxylic acid via the epoxide intermediate connection.

Thus, the interaction of the third compound with an oxidizing agent may be carried out by transformation of unsaturated groups in the epoxide and the conversion of the epoxide to carboxylic acid. Preferably the conversion of the epoxide to carboxylic acid is carried out by conversion of the epoxide to diol and diol oxidation to obtain a carboxylic acid group.

The transformation of the third connection epoxide is preferably carried out by the interaction of the third connection with the peroxide, as described above, and more preferably W is ccarbonate acid.

The conversion of the epoxide to diol is preferably carried out by the interaction of the epoxide with perchloro acid (HClO4). Alternatively, epoxide converted into diol interaction with sulphuric acid.

Diol is then converted into carboxylic acid by interacting with the third oxidizing agent. Third oxidizing agents can be selected from periodate, bismuthate, permanganate, chlorite, and any combination thereof. Preferably diol converted into carboxylic acid by interaction with periodate, with subsequent interaction with chlorite.

Thus obtained fourth connection with glycerin main chain and containing carboxylic acid portion that is attached to it through a simple radio link, then purified to obtain the purified product.

The authors of the present invention in this work, suddenly discovered that the fourth connection received through the epoxy intermediate connection can be easily cleaned at the same time to exclude the use of column chromatography, if its free hydroxyl group protected by a protective group, such as acetate, pivaloate or benzoate.

As mentioned here above, the free hydroxyl group, if it is present in glycerol main chain, preferably protects, while preferably the th, selective protecting group is trityl. However, as trityl is big, bulky and non-polar group, its presence may, in some cases, complicate the methods of isolation and purification of intermediates of the synthesis and the final product.

The inventors in the present work found that the limitations associated with trailvoy group, can be overcome: (i) allocation aldehydebase compounds through the formation of the water-soluble adduct, as broadly described herein above; or (ii) replacement trailvoy protective groups are less bulky group, after the introduction of the second connection. In addition, as indicated here above, when the oxidation of the third connection includes education diol, after the formed diol, trailing protective group can be removed and the resulting triol can be distinguished by means of crystallization.

Thus, according to a preferred variant implementation of the present invention the method further comprises after receiving the purified third compound and/or to the interaction of the third compound with an oxidizing agent replacement trailvoy group with a protective group selected from the group consisting of acetate, pivaloate or benzoate.

Replacement trailvoy protective groups are usually carried out by removing trailvoy GRU is dust so to get a free hydroxyl group, protected hydroxyl group required protective group.

Protection of the hydroxyl group of the acetate group is easily carried out by the interaction of the third connection, for example, with acetic anhydride. Protection of the hydroxyl group pivaloate group is easily carried out by the interaction of the third connection, for example, revalorisation. Protection of the hydroxyl group benzoate group is easily carried out by the interaction of the third connection, for example, benzoyl chloride.

Cleaned fourth connection as described here, which has a acetate, pivaloate or benzoate protective group, you can perform the conventional methods of extraction, preferably by use of silica gel during the process of extraction.

As described in the examples section that follows (see example 2), found that obtaining glycerin compounds having containing oxidized group, a group attached through an ether linkage, through education epoxydodecane intermediate compound, which has the acetate protective group, leads to vysokoochishchennogo connection and high output.

In cases where the received fourth connection has a protective group, as described herein above, after the fourth connection is received and cleared out, the protective group is removed.

When oxidized group is an ester, the method is carried out by obtaining compounds containing carboxylic acid, and then converting the carboxylic acid in ether. This can easily be done using methods well known in the field.

As discussed here above, compounds having glycerin main chain and at least one residue containing oxidized group attached to the glycerol main chain, and also having including a phosphorus-containing group, a part attached to glycerol main chain, preferably phosphate-containing part, known as oxidized phospholipids and are very useful in the treatment of various conditions. Thus, the method described herein, optionally and preferably, also includes the introduction of a part comprising the phosphorus-containing group, glycerin main chain.

Used here, the phrase "part, including phosphorus-containing group, describes the part as shown here, which includes one or more phosphorus atoms. Representative examples include, without limitation listed, phosphates, phosphonates, phosphines, phosphine oxides, phosphites, pyrophosphates and the like.

Used here, the term "phosphonate" describing the t group-P(=O)(OR')(OR"), where R' and R"each independently represents hydrogen or substituted or unsubstituted alkyl, cycloalkyl or aryl, as described here.

The term "phosphinyl" describes a group-PR R', where R' and R" have the meanings specified above.

The term "phosphine oxide" refers to the end group-P(=O)(R')(R") or-P(=O)(R')-linking group, as these phrases are listed here above, where R' and R" have the meanings as indicated above.

The term "pyrophosphate" describes a group-O-P(=O)(OR')-O-P(=O)(OR')(OR")(OR"')where R', R" have the above meanings and R"' have the meanings as R' or R".

The term "postit" describes a group-O-PH(=O)(OR')where R' has the meanings specified above.

The term "phosphate" refers to the group-O-P(=O)2(OR')where R' has the meanings specified above.

The term "thiophosphate" describes a group-O-P(=O)(=S)(OR')where R' has the meanings specified above.

Introduction part comprising phosphorus-containing group, glycerin connection can be made to interact with the first connection and the second connection, to the selection of the third connection to the interaction of the third compound with an oxidizing agent, prior to the allocation of the fourth connection or after allocation of the fourth connection, and can be produced using any method known in this field.

Introduction part comprising phosphorus-containing group, in connection with glycerine the e connection therefore, carry out:

the interaction of the first connection, the third connection, the purified third compound, a fourth compound or purified fourth compound described above, with the part which includes a phosphorus-containing group, so as to obtain a connection with glycerin main chain and at least one residue containing oxidized group attached to the glycerol main chain and, in addition, including having phosphate group part attached to glycerol main chain.

According to a preferred variant implementation of the present invention includes a phosphorus-containing group is part represents the phosphate part that is attached to glycerol main chain through postradiation connection.

Thus, the part which includes a phosphorus-containing group may be, for example, phosphoric acid, phosphorylcholine, phosphorylethanolamine, fosfomycin, phosphorylation, fosforilmetil, ethylphosphonic, phosphorimetry, phosphorylation, fosforilirovanii, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphorylethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphonate, pyrophosphate, phosphatidylethanolamine, dinitrobenzophenone and fosfoglitserin.

Preferably the art, includes a phosphate group attached at the sn-3 position of glycerol main chain and therefore the introduction of such parts is carried out selectively, as appropriate, the protection of other free hydroxyl groups that are present in the reactive compound or the removal of the protective group in the protected hydroxy-group in the desired position.

In currently known methods of production of oxidized phospholipids comprising phosphorus-containing group part is usually injected to obtain compounds containing oxidized group.

In addition, in cases where the part that includes phosphorus-containing group is phosphorylcholine, widely used and useful part in such compounds currently known methods include the reaction of N-alkylation involving hazardous and noxious environmental agents, such as, for example, trimethylamine.

The inventors in the present work found that (i) the part that includes phosphorus-containing group, you can easily enter after obtaining compounds containing oxidized group; and (ii) implementation introduction part comprising phosphorus-containing group, can now be done through the formation of an intermediate product comprising a reactive phosphorous-containing group.

On the basis of the above authors, and the gain has developed and successfully implemented in practice a new way of introduction part, having a phosphate group, in connection with glycerin main chain and containing oxidized group balance, prisoedinenii to it through a simple radio link.

This method, combined with the above-described method for obtaining compounds containing oxidized group, can advantageously be used to obtain therapeutically useful oxidized phosopholipid described above.

Thus, according to preferred variants of implementation of the present invention the introduction part, with the phosphorus-containing group, carried out after receipt of the third connection, or after receiving the fourth connection, the latter is preferred. However, it should be noted that the method of introducing part having a phosphate group, is also suitable for any other stage.

Introduction part comprising phosphorus-containing group, glycerin connection, therefore, preferably carried out under the proposed options for the implementation of the interaction of the first connection, the third connection, the purified third compound, a fourth compound or purified fourth compound, as described above, which has a free hydroxyl group, with a reactive compound comprising phosphorus-containing groups which, to obtain a compound having a reactive group comprising phosphorus-containing group; and the transformation of the reactive group comprising phosphorus-containing group, in part, with a phosphorus-containing group.

Reactive compound comprising phosphorus-containing group, are chosen so that when the above-mentioned interaction to obtain a phosphorus-containing group, a reactive group, prisoedineniu to glycerol main chain. Reactive compound comprising phosphorus-containing group, therefore, choose one that has a second reactive group and a third reactive group and it will choose the second reactive group capable of interacting with a hydroxyl group, and choose the third reactive group capable of becoming a part having the phosphate group.

Reactive groups that are capable of interacting with the free hydroxyl groups include, for example, halides, sulphonylchloride, acylhomoserine and the like.

Preferably, the second reactive group is a halide, and more preferably it is a chloride.

When, as indicated above, the preferred parts, including phosphorus-containing group, are phosphate cha is ti, the transformation of compounds having a phosphate group in the desired portion having a phosphate group, usually involves the formation of phosphate broadcasting communication. Such communication can be obtained, for example, the interaction derivative of phosphoric acid, such as phosphorylchloride with the part containing the hydroxy-group.

Thus, according to a preferred variant implementation of the reactive compound comprising phosphorus-containing group is phosphorous oxychloride (POCl3), so that the third and the second reactive group, both are the chlorides, and the compound having a reactive group comprising phosphorus-containing group, has glycerin main chain and attached to it phosphorylchloride the rest.

The interaction of the first connection, the third connection, the purified third compound, a fourth compound or purified fourth compound with phosphorous oxychloride is usually carried out in the presence of a base. Suitable bases include organic and inorganic bases and organic bases are preferred. Thus, the reaction is preferably carried out in the presence of a base, such as, for example, trialkylamine (e.g., triethylamine).

This reaction, in addition, carry out the in the presence of a solvent, preferably polar solvent, such as THF.

Phosphorylchloride, piaristengasse compound obtained by the method mentioned above, can easily be turned in any desired part, including phosphorus-containing group, and it is, therefore, a useful intermediate product.

For example, it can be converted into phosphoric acid by simple hydrolysis, as described in the example in the examples section that follows below.

Alternatively, it can communicate with the part containing the hydroxy-group, and optionally and preferably also with water to get through this other phosphate parts.

Preferred phosphate parts that enter into therapeutic oxidized phospholipids (for example, phosphorylcholine, phosphorylethanolamine) usually contain aminoalkyl group, which, moreover, can be N-alkilirovanny.

Turning phosphorylchloride intermediate product in phosphate such parts can thus easily be carried out by the interaction with the derived desired aminoalkyl group chosen as being able to interact with a third reactive group (which chloride).

For example, aminoalkylsilane phosphate part can be obtained by the interaction phosphorylchloride Prohm is filling product with linesperson. If you want, amerosport can then be further alkilirovanii to obtain N-alkilirovanny aminoalkylphosphonic part, as in the case of phosphorylcholine part.

The receipt of such N-alkilirovanny aminoalkylphosphonic part attached to glycerol main chain, using the method specified above, is very advantageous because it eliminates the use of hazardous substances, such as trimethylamine, usually used for obtaining such compounds.

As indicated above, the introduction part comprising phosphorus-containing group can be carried out either before or after the introduction of the residue containing the oxidized group in glycerol connection. As described in the examples section that follows below, phosphorylcholine part was successfully introduced in glycerol compounds having either a residue containing oxidized group, or a residue containing an unsaturated group (see examples 4 and 5). Thus, the method of introduction of phosphate through the reactive intermediate compound having a phosphorus-containing group, proposed here, can be done with glycerin compounds having either oxidized or pre-oxidized group attached thereto through an ether linkage. When searching for improved ways to receive the deposits of oxidized phospholipids authors of the invention, in addition, developed and implemented in practice more glycerin compounds with oxidized group attached to it through a simple radio communication which is performed by the direct introduction of the residue containing the oxidized group in glycerol connection.

On this basis, according to another aspect of the present invention, a method for obtaining compounds with glycerin main chain and at least one residue containing oxidized group attached to the glycerol main chain through an ether linkage, which carry out:

obtaining a first connection with glycerin main chain and at least one free hydroxyl group as described above;

obtaining a fifth connection having at least one oxidized group, as defined above, and at least one fourth reactive group;

the interaction of the first connection and the fifth connection obtained through this reaction mixture, containing the sixth connection which is the connection with glycerin main chain and at least one containing oxidized group, residue, attached to glycerol main chain through an ether linkage; and

allocating connection with glycerin main chain and at least one containing oxidized group, residue, attached to glycerol main chain through an ether linkage.

The method according to this aspect of the present invention, therefore, includes the interaction of the first connection specified above, with a compound which has a reactive group that is capable of interacting with a free hydroxyl group of the first connection, shown here as a fourth reactive group, and oxidized group. This connection is designated here as the fifth connection.

Oxidized group at the fifth connection can be any oxidized groups specified above, namely aldehyde, diol, carboxylic acid, ester, acetal and ketal. Optional, oxidized group can be polyarylene group, and it is easily transformed into the desired oxidized group without interaction with the oxidizing agent. An example of such polyarylene group is a nitrile, which can be easily converted to carboxylic acid by simple hydrolysis.

Fourth reactive group in the fifth connection has the values specified for the first reactive group, and is preferably a halide and more preferably a bromide.

The interaction of the first connection and the fifth connection is preferably carried out in the presence of the warping. Preferred are relatively strong inorganic bases, such as, for example, sodium hydride, sociallyengaged, sodium amide, sodium hydroxide and any mixture.

Under such reaction conditions, the fifth connection that has 4 or 5 carbon atoms, can undergo cyclization during the reaction, thereby adversely affecting the efficiency of the reaction.

Thus, preferably the fifth connection preferably has less than 4 or more than 5 carbon atoms.

As described above, when the first connection has more than one attached to a hydroxyl group, a hydroxyl group optionally and preferably protects the protective group to interaction of the first and fifth connections.

After you receive the sixth connection, the protective group can be removed and the connection is cleaned using conventional cleaning methods.

The method according to this aspect of the present invention is very advantageous, since it allows to obtain said compound containing oxidized group, one-step synthesis.

According to this method, the oxidized phospholipids can be easily obtained by the introduction part comprising phosphorus-containing group, as described above, either before or after interaction with the fifth connection. the conducting part, includes a phosphorus-containing group, preferably carried out using the method specified above.

In any of the ways listed above, the first connection may include attached alkylenes chain. Preferably Allenova chain attached at the sn-1-position of the first connection.

Allenova circuit may be joining glycerin connection, for example, ester bond or a simple ester bond. Preferably Allenova chain attached through ether linkage so that the end product is glycerin connection with two groups of simple ether.

Thus, in each of the ways listed above, the first compound is glycerolipid, as stated here, and preferably having one group of simple ether glycerolipid, in which the lipid portion attached at the sn-1 position of glycerol. This connection therefore has one free hydroxyl group, which, as indicated above, preferably protect before any interaction.

Thus, the first compound may be, for example, glycerin, glycerolipid, glycerolipid with one group of simple ether, glycerolipid two groups of simple ether, fosfoglitserin, phosphoglyceric transformed into simple monoether phosphoglyceric and lysolecithin.

<> As discussed in detail above, the position in which the residue containing oxidized group attached to the glycerol main chain, affect the activity of the compounds obtained and, thus, as further discussed above, it is preferable to conduct the reaction selectively.

Preferably in any of the ways specified above, the remainder containing oxidized group attached at the sn-2-position of the connections. So accordingly the selection and/or protection of the first connection provide selective connection of the residue containing the oxidized group.

Therefore, in the preferred embodiment of the present invention, the first compound has the following General formula I:

The formula I

in which:

And1absent or selected from the group consisting of CH2, CH=CH and C=O;

R1selected from the group consisting of H and hydrocarbon chain having 1-30 carbon atoms; and

R3selected from the group consisting of hydrogen, alkyl, aryl, phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, phosphorylate the Ola, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine, fosfoglitserin.

Using any of the methods described above, it is possible thus to obtain a compound having the following General formula II:

Formula II

in which:

And1selected from the group consisting of CH2, CH=CH and C=O, and preferably represents CH2;

And2represents CH2;

R1represents alkyl having 1-30 carbon atoms;

R2is

where X is an alkyl chain having 1-24 carbon atoms;

Y is selected from the group consisting of hydrogen, hydroxy, alkyl, alkoxy, halogenide, acetoxy and aromatic functional groups; and

Z is selected from the group represents:

where R4is alkyl or aryl; and

R3selected from the group consisting of hydrogen, alkyl, aryl, phosphoric acid, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, FOS is allcation, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine, fosfoglitserin.

Additional objectives, advantages and new features of the present invention will be obvious to any average person skilled in the art after consideration of the following examples which are not intended to limit the invention. In addition, each of the various embodiments and aspects of the present invention, as described above and as indicated in the claims below finds experimental confirmation in the following examples.

EXAMPLES

In this paper links to the following examples, which together with the description presented above illustrate the invention but do not restrict it.

General synthetic route:

Under the guidance of the present invention to obtain oxidized phospholipids used some common synthetic concept, as follows below:

(i) obtaining glycerolipids compounds having at least one containing oxidized group mod is to, attached through ether bond, attach unsaturated residue to glycerolipid and oxidation of unsaturated communication, using reagent Gerard and/or crystallization trilateral connections for separation of the oxidized product, as shown in Example 1 and the Schemes I-V;

(ii) obtaining glycerolipids compounds having at least one containing oxidized group residue attached via ether bond, attach unsaturated residue to glycerolipid and oxidation of unsaturated communication through the epoxy intermediate connection with the use of protective acetochlor, as shown in example 2 and schemes VI-X;

(iii) obtaining glycerolipids compounds having at least one containing oxidized group residue attached through ether linkage direct introduction compounds containing oxidized group, as shown in example 3 and figure XI; and

(iv) the introduction of the reaction part comprising phosphorus-containing group, glycerolipids connection having one or two containing oxidized (or pre-oxidized) group residue attached through ether linkage, using reactive compound comprising phosphorus containing group (e.g., phosphorous dichloride), for the formation of actiongo intermediate compounds, as shown in examples 4 and 5 and on the schemes XII-XIV.

EXAMPLE 1

Getting rat(racemic)- 1-hexadecyl-2-(5'-methyl ether of pentanol acid)-glycerol using periodate and reagent Gerard

In this example, the unsaturated group introduced in glycerol main chain and then oxidized by means of formic acid, hydrogen peroxide and periodate. Then formed in this way the oxidized product was purified using a reagent Gerard.

As an illustrative example described here getting rat-1-hexadecyl-2-(5'-methyl ether of pentanol acid)and glycerin.

1-Hexadecyl-2-(5'-methyl ether of pentanol acid)-glycerol was obtained according to the methodological instructions of the present invention, as shown in schemes I-V below.

1-Hexadecyl-3-trailerin received, as described in U.S. patent No. 6838452. Briefly, D-acetoglyceride (4 g), powdered potassium hydroxide (approximately 10 grams) and hexadecylamine (9.3 grams) in benzene (100 ml) was stirred and boiled under reflux for 5 hours with removal of water formed by azeotropic distillation (compare W.J. Baumann and H.K.Mangold, J. Org. Chem. 29: 3055, 1964 and F.Paltauf, Monatsh. 99:1277, 1968). The volume of solvent was gradually reduced to approximately 20 ml, and the resulting mixture was cooled to room temperature and was dissolved in ether (100 is l). The resulting solution was washed with water (250 ml), and the solvent was removed under reduced pressure. To the residue was added 100 ml of a mixture of 90:10:5 methanol:water:concentrated hydrochloric acid and the mixture is boiled under reflux for 10 minutes. The product was extracted with ether (200 ml) and washed successively with water (50 ml), 10% sodium hydroxide (20 ml) and again water (volume 20 ml) until neutral indicator. Retorical was removed under reduced pressure, and the product (8.8 g) was led from hexane to obtain 7.4 g of pure 1-hexadecylamine.

1-Hexadecyloxypropyl to (7.9 grams), triphenylmethane (8.4 grams) and dry pyridine (40 ml) was heated at 100C for 12 hours. After cooling, was added 300 ml of ether and 150 ml of ice water and the reaction mixture was transferred into a separating funnel. The organic phase is washed successively with 50 ml ice water, 1% potassium carbonate solution (up to an alkaline value) and 50 ml of water, then dried over anhydrous sodium sulfate. The solvent is evaporated, the residue was dissolved in 150 ml of warm petroleum ether, and the resulting solution was cooled at 4C over night. After separation of the precipitate by filtration, the filtrate evaporated, and the residue was recrystallized from 20 ml of ethyl acetate at -30C receives 8.2 grams of 1-hexadecyl-3-tritylglycine, melting point 49C.

As shown With the EME I, 1-hexadecyl-3-trailerin (14,78 grams, 0,0265 mol), 6-bromo-1-hexene (4,85 g) and powdered potassium hydroxide (approximately 10 grams) in hexane (200 ml) was stirred and boiled under reflux for 6 hours with removal of water formed by azeotropic distillation. The reaction mixture was cooled to room temperature, washed with water (3100 ml), the solvent was removed under reduced pressure. The residue was dissolved in chloroform (50 ml) and was purified by filtration through silica gel 60 (12.5 grams). The chloroform was removed under reduced pressure, and the residue was dissolved in petroleum ether (100 ml). The solution was kept at 4C during the night, this was accompanied by the deposition of by-products. After filtration and removal of solvent under reduced pressure received 12,15 grams (0,0190 mol) 1-hexadecyl-2-(5'-hexenyl)-3-tritylglycine (yield 72%).

Scheme I

1-Hexadecyl-2-(5'-hexenyl)-3-trailerin (19,80 grams) was dissolved in formic acid (100 ml). The yellow solution was stirred at room temperature for 2 hours and then cooled in an ice bath. Hydrogen peroxide 33% (25 ml) was added dropwise to ice the solution for 50 minute the reaction mixture immediately changed from yellow to white. When you are finished adding continued stirring in an ice bath on the additional 4 hours. Then the reaction mixture was poured into ice (150 g) and was extracted with ether (3100 ml). Essential orange solution was washed with water (100 ml), and the solvent was removed under reduced pressure. The residue was dissolved in dichloromethane (150 ml), washed with saturated aqueous sodium bicarbonate (100 ml), and the solvent was removed under reduced pressure. Then the residue was dissolved in hot hexane (250 ml). Immediately place the deposition of the white connection. The solution was kept at 4C over night. After the precipitate (0.53 g). filtration followed by removal of solvent under reduced pressure received 20,03 grams of a yellow oily residue. This residue was dissolved in isopropanol (200 ml) was added an aqueous solution of sodium hydroxide (17 g in 50 ml water). The resulting solution was heated to 90C for 2 hours and then cooled and poured into ice (150 grams). Then the mixture was extracted with dichloromethane (3100 ml), the organic phase is washed with water (100 ml) and saturated aqueous solution of sodium dihydrophosphate and dried over anhydrous Na2SO4. After removal of the solvent under reduced pressure received 10,77 grams of the crude product. Then the crude product was dissolved in 80% methanol (100 ml)and the solution kept at 4C over night. Was separated by filtering the precipitate and removed most of the solvent is ri reduced pressure. Was extracted with dichloromethane (3100 ml), dried over anhydrous Na2SO4and solvent was removed under reduced pressure. Paracrystalline from hexane (250 ml) was received 7,44 grams of pure 1-hexadecyl-2-(5',6'-dihydroxyethyl)glycerol.

Scheme II

As shown in scheme III, 1-hexadecyl-2-(5',6'-dihydroxyethyl)glycerin (7,84 grams) was dissolved in isopropanol (50 ml) and water (12 ml). Added NaIO4(9 grams) and stirred the reaction mixture at room temperature for 3 hours. Was added water (50 ml) and the reaction mixture is extracted with chloroform (350 ml), dried over anhydrous Na2SO4, was filtered, and the solvent was removed under reduced pressure, getting to 5.56 grams. The crude product was dissolved in ethanol (60 ml) and glacial acetic acid (2.3 g). Added reagent Gerard (5.6 grams), and the reaction mixture is boiled under reflux for 2 hours. The reaction mixture was cooled in an ice bath, was added a solution of alkali (2.3 g in 45 ml water) and the mixture was extracted with ether (325 ml). The ether phase is washed with water and the water combined with the alkaline phase. The aqueous phase was acidified with concentrated HCl (4.4 ml) and was extracted with ether (325 ml). Washed with water, saturated aqueous sodium bicarbonate (325 ml), water (225 ml), dried over anhydrous Na2SO 4and solvent was removed under reduced pressure, received 1,95 grams (0,0049 mol) 1-hexadecyl-2-(5'-oxapentane)glycerin (output 26,9%).

Scheme III

As shown in scheme IV, 1-hexadecyl-2-(5'-oxobutyl)glycerin (4,80 grams) was dissolved in dry triethylamine (57 ml). Was added acetic anhydride (20 ml) and the reaction mixture was stirred at room temperature for 2.5 hours. The reaction mixture was poured into ice (100 ml) and was extracted with dichloromethane (3100 ml). The organic phase was washed successively with water (100 ml), dilute hydrochloric acid (100 ml), water (100 ml), saturated aqueous sodium bicarbonate (100 ml) and again water (100 ml)and then dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain 4.54 grams of 1-hexadecyl-2-(5'-oxobutyl)-3-acetate of glycerol (yield 86%).

1-Hexadecyl-2-(5'-oxobutyl)-3-acetate of glycerol (3.94 g) was dissolved in tert-butanol (75 ml). The sodium chlorite (6,85 grams) and the dihydrate of sodium dihydrophosphate (15,50 grams) was dissolved in water (75 ml). The aqueous solution was added to the alcohol solution, and the reaction mixture was stirred at room temperature for 4 hours. Then the reaction mixture was transferred into a separating funnel and was extracted with dichloromethane (3100 ml). The combined organic phase was washed with water (2100 ml), and RA is the solvent was removed under reduced pressure. The residue was dissolved in a mixture of methanol (80 ml) and 10% aqueous NaOH (20 ml)and the solution was stirred at room temperature overnight. The methanol solution was extracted with a mixture of toluene and hexane (1:1) (250 ml), cooled in an ice bath and slowly added conc. HCl to pH 5-6. Then the solution was extracted with dichloromethane (2100 ml). The combined organic phase was washed with water (100 ml), dried over anhydrous Na2SO4and solvent was removed under reduced pressure to get 2,07 grams of the crude product. By recrystallization from hexane (20 ml) was received of 1.30 grams of pure 1-hexadecyl-2-(4'-carboxy)butylamine (yield 35%).

Scheme IV

As shown in scheme V, to the residue was added methanol (100 ml) and 10% aqueous NaOH (20 ml)and the resulting solution was stirred at room temperature for 2 hours. The solution was extracted with a mixture of petroleum ether/toluene (1:1, V/V), and methanol phase was acidified to pH 0 with concentrated HCl and then was extracted with chloroform (350 ml). The combined chloroform phase was washed with water (20 ml), dried over anhydrous Na2SO4, and the solvent was removed under reduced pressure, getting 0.77 grams (0,00179 mol) rat-1-hexadecyl-2-(5'-methyl ether of pentanol acid)glycerol (96,7%).

Scheme V

EXAMPLE 2

Recip is the rat-1-hexadecyl-2-(5'-methyl ether of pentanol acid)glycerol using periodate and acetate protective group

In this example, the unsaturated group introduced in glycerol main chain and then oxidized to an ester via epoxide using acetic anhydride, 4-chlorobenzotrichloride, HClO4, periodate and methanol. Efficient allocation of intermediate products was carried out by conducting reactions with the use of acetate protective group.

As an illustrative example described here getting rat-1-hexadecyl-2-(5'-methyl ether of pentanol acid)glycerol.

Rat-1-hexadecyl-2-(5'-methyl ether of pentanol acid)glycerol was obtained according to the methodological instructions of the present invention, as described in schemes VI-X, below.

As shown in scheme VI below, 1-hexadecyl-2-(5'-hexenyl)-3-trailerin, obtained as described in example 1 above (4,90 g), was dissolved in a mixture of methanol (30 ml) and conc. hydrochloric acid (3 ml)and the resulting solution was heated to boiling under reflux for 4 hours. The reaction mixture was cooled to room temperature, poured into ice (100 ml) and was extracted with chloroform (3100 ml). The organic phase is washed with water (100 ml), aqueous sodium bicarbonate (100 ml) and again water (100 ml). Then the organic phase was dried over anhydrous Na2SO4was filtered and solvent was removed, receiving a 3.75 grams of residue. Osteoclastoma in n-hexane and kept at 4C over night. After separation by filtration of sediment and removal of solvent received 3,17 grams of product which was dissolved in chloroform (200 ml) and was added to silica gel (45 grams). This solution was filtered and the silica gel was extracted again with a mixture of chloroform:methanol (200 ml, 9:1) and chloroform:methanol (200 ml, 1:1). The last two extracts were combined and the solvent was removed under reduced pressure to obtain of 2.56 grams of 1-hexadecyl-2-(5'-hexenyl)-glycerol (yield 84%).

Scheme VI

As shown in scheme VII below, the dry pyridine (5 ml) and acetic anhydride (3 ml) was added to the obtained 1-hexadecyl-2-(5'-hexenyl)glycerol and heated the reaction mixture at 70C for 2 hours. The reaction mixture was poured on ice (25 g) and was extracted with hexane (325 ml). The extract was washed successively with water (25 ml), aqueous diluted sulfuric acid (25 ml), water (25 ml), aqueous sodium bicarbonate (25 ml) and water. After drying over anhydrous Na2SO4, filtration and removal of solvent received 2,60 grams. The residue was dissolved in dichloromethane (50 ml) was added 3-chloroperbenzoic acid (3,84 grams), and the reaction mixture was stirred at room temperature overnight. Reduced the amount of solvent under reduced pressure, leaving approximately 20 ml, and was added n-hexane (100 ml). After the filtrowanie the solvent evaporated to dryness. The residue was dissolved in n-hexane (100 ml)was added a solution of alkali (0.4 g NaOH in 50 ml water), and the phases were separated. Washing of the organic phase successively with water (25 ml), aqueous sodium bicarbonate (25 ml), water (25 ml), drying over anhydrous Na2SO4, filtration and removal of solvent received 2,40 grams of 1-hexadecyl-2-(5',6'-epoxyhexane)-3-acetate of glycerol (yield 82%).

Scheme VII

As shown in scheme VIII below, 1-hexadecyl-2-(5',6'-epoxyhexane)-3-acetate of glycerol was dissolved in acetone (50 ml). Added 7% HClO4(5 ml)and the reaction mixture was stirred at room temperature for 40 hours. Was added water (50 ml)and the reaction mixture was extracted with chloroform (350 ml). Washing of the organic phase successively with water (25 ml), aqueous sodium bicarbonate (25 ml), water (25 ml), drying over anhydrous Na2SO4, filtration and removal of solvent received to 2.29 g of oily residue. The residue was dissolved in chloroform (200 ml) and was added to silica gel (30 g). This solution was filtered, and the silica gel was again extracted with a mixture of chloroform:methanol (200 ml, 8:2). From the second extract after removal of the solvent under reduced pressure got to 1.45 grams of 1-hexadecyl-2-(5',6'-dehydroretinal)-3-acetate of glycerol.

Scheme VIII

As shown in Scheme IX below, 1-hexadecyl-2-(5',6'-dihydroxyethyl)-3-acetate of glycerol was dissolved in isopropanol (50 ml). Was added an aqueous solution of periodate sodium (1,45 g in 50 ml water)and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was extracted with chloroform (350 ml), dried over anhydrous Na2SO4, was filtered, and the solvent was removed under reduced pressure, getting 0.96 grams. The residue was dissolved in tert-butanol (50 ml), was added an aqueous solution (50 ml) of sodium chlorite (of 1.66 grams) and the dihydrate of sodium dihydrophosphate (3,76 grams). The reaction mixture was stirred at room temperature for 4 hours, was extracted with chloroform (250 ml), and the solvent was removed under reduced pressure. The residue was dissolved in a mixture of chloroform:hexane (200 ml, 1:1) was added to silica gel (15 grams). The solution was filtered, and the silica gel was extracted again with chloroform (200 ml) and a mixture of chloroform:methanol (200 ml, 9:1). The solvent from the last extract was removed under reduced pressure, getting to 0.92 grams of 1-hexadecyl-2-(4-carboxybutyl)-3-acetate of glycerol.

Scheme IX

As shown in figure X below, 1-hexadecyl-2-(4'-carboxybutyl)-3-acetate of glycerol was dissolved in 50 ml of a mixture of 8:2 methanol and 10% aqueous NaOH, and the reaction mixture energy is a rule was stirred at room temperature overnight. The reaction mixture was extracted with a mixture of toluene:petroleum ether (225 ml, 1:1). The methanol phase was acidified with concentrated HCl to obtain a pH of approximately 0 and then was extracted with chloroform (225 ml). The solvent was removed under reduced pressure, and the residue was dissolved in methanol (10 ml). Added concentrated HCl (2 drops)and the solution was stirred at room temperature overnight, then was extracted with chloroform (225 ml), followed by washing the organic phase with water (25 ml), then washing with aqueous sodium bicarbonate (25 ml), water (25 ml), drying over anhydrous Na2SO4, filtration and removal of solvent, receive 0,86 gram of pure rat-1-hexadecyl-2-(5'-methyl ether of pentanol acid)glycerol.

Scheme X

EXAMPLE 3

Getting rat-1-hexadecyl-2-(5'-ethyl ether of pentanol acid)glycerol direct introduction of oxidized groups

rat-1-Hexadecyl-2-(5'-ethyl ether of pentanol acid)glycerol was obtained according to the methodological instructions of the present invention, as shown in scheme XI below.

Scheme XI

1-Hexadecyl-3-trailerin received, as indicated, for example, in example 1 above or as described in U.S. patent No. 6838425.

In a three-neck flask with magnetic is th stirrer was added 1.0 g (1.8 mmol) of 1-hexadecyl-3-tritylglycine, 0,78 g (3.6 mmol) ethyl ester 5-bombalurina acid and 75 ml of dimethylformamide (DMF). To mix the solution was added dropwise to 0.20 grams (5 mmol) of NaH (60% dispersion in mineral oil)dissolved in 25 ml of dimethylformamide for 15 minutes, and stirring was continued for additional 1 hour to complete the reaction. Was added water (50 ml) and the mixture was extracted with ether (350 ml). The organic phase was dried over anhydrous Na2SO4, and the solvent was removed under reduced pressure. The crude product was purified column chromatography on silica gel.

Removing the protective trailvoy group, as described above, received the final product.

EXAMPLE 4

Introduction part comprising phosphorus-containing group, glycerolipids connection

Under the guidance of the present invention, the reaction part including a phosphorus-containing group is introduced into glycerolipids connection having one or two containing oxidized (or pre-oxidized) group residue attached through ether linkage. Introduction reactive part comprising phosphorus-containing group, carried out with the use of compounds comprising phosphorus-containing group, such as, for example, phosphorous oxychloride. Optionally, subsequent to the introduction of the Rea is cyanopogon part, includes a phosphorus-containing group, it turned to the phosphate part.

Getting rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-dichlorophosphate

As an illustrative example, the rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-dichlorophosphate received under the guidance of the present invention, as shown in scheme XII below.

Scheme XII

Thus, of 0.24 ml (0,39 grams of 2.53 mmol) POCl3and 10 ml of tetrahydrofuran (THF) were placed in a cooled ice three-neck flask with a magnetic stirrer. To mix the solution was added dropwise over 25 minutes, a mixture of 0.87 g (2.2 mmol) rat-1-hexadecyl-2-(5'-hexenyl)glycerol, 0,34 ml (0.25 grams of 2.44 mmol) of triethylamine and 50 ml of tetrahydrofuran (THF)and stirring was continued for additional 10 minutes in a bath with ice and then continued for 45 minutes at 23C.

rat-1-Hexadecyl-2-(5'-hexenyl)-glycero-3-dichlorophosphate can hydrolyze to obtain thereby the corresponding fosfatados acid, as follows below:

to the reaction mixture was added 1 gram of ice and continued stirring for 30 minut.zatem was added water (50 ml) and product was extracted with a mixture of chloroform:Meon (2:1, V/V, 325 ml). The organic phase is washed with water, and the solvent was removed under reduced pressure.

In the alternative is ariante rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-dichlorophosphate can interact with various derivatives of the alkylamine to get through this phosphoglyceride, as shown below.

Getting rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-phosphoethanolamine from rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-dichlorophosphate

rat-1-Hexadecyl-2-(5'-hexenyl)-glycero-3-phosphoethanolamine was obtained according to the methodological instructions of the present invention, as shown in scheme XIII

Scheme XIII

A solution of rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-dichlorophosphate in THF obtained, as indicated here above, in example 2, was cooled in a bath with ice. To the solution was added dropwise during 10 minutes to a mixture of 0.16 ml (0.16 g, 2.7 mmol) ethanolamine, 0,34 ml (0.25 g, 2.4 mmol) of triethylamine and 50 ml of THF. After you have added all the solutions, the resulting solution was stirred additionally for 20 minutes and then removed the bath with ice and stirred over night at room temperature.

The solution was filtered using filter paper (Whatman #2). The residue remaining on the filter paper, dried under reduced pressure, obtaining 1.2 g of the residue is not quite white.

1.2 grams of the residue is not completely white was dissolved in a mixture of 24 ml of glacial acetic acid and 10 ml of water, kept at 70C for 1 hour and allowed to cool to room temperature. The product was extracted from the acetic acid solution fed is owned by washing with 50 ml of solution for extraction with chloroform:methanol 2:1. The solvent solution for the extraction evaporated, getting 0.94 g (1.7 mmol) rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-phosphoethanolamine, exit 85% compared to rat-1-hexadecyl-2-(5'-hexenyl)glycerol.

Getting rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-phosphocholine from rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-phosphoethanolamine

rat-1-Hexadecyl-2-(5'-hexenyl)-glycero-3-phosphocholine received under the guidance of the present invention, as shown in scheme XIV below

Scheme XIV

In a three-neck flask with magnetic stir bar was added to 0.50 g (0,99 mmol) rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-phosphoethanolamine, 50 ml isopropanol and 18 ml of CH2Cl2. During the stirring was added a mixture of 5 grams To2CO3and 10 ml of water, and the solution temperature was maintained between approximately 35and approximately 40C, while added dropwise a mixture of 1.0 ml (1.3 g, 11 mmol) dimethylsulfate and 10 ml isopropanol for 45 minutes. After adding all of the solution the solution was stirred additionally for 90 minutres was allowed to cool to room temperature. The product was extracted from the solution three times washing with 50 ml of a mixture of chloroform:methanol 2:1. The solvent solution is evaporated, thus obtaining 0,50 g (0.82 mmol) rat-1-

from what oseney to rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-phosphoethanolamine.

The purity was confirmed by thin-layer chromatography on aluminum oxide using as elution solvent a solvent mixture of chloroform:methanol: water (70:26:4). The identity of the rat-1-hexadecyl-2-(5'-hexenyl)-glycero-3-phosphocholine confirmed using13C-NMR.

EXAMPLE 5

Getting 1-hexadecyl-2-(5'-carboxymethyl)butyl-glycero-3-phosphocholine

A solution of 1-hexadecyl-2-(4'-carboxymethyl)butylparaben (0,86 g), 0,34 g (2.6 mmol) of triethylamine and 50 ml of tetrahydrofuran was added dropwise over 25 minutes to a cooled with ice to a solution of 0.24 ml (0,39 g, 2.6 mmol) POCl3and 10 ml of tetrahydrofuran (THF). The resulting mixture was stirred additionally for 10 minutes in a bath with ice and within 45 minutes at room temperature (23C). Then the reaction mixture was cooled in a bath with ice, and thereto with vigorous stirring solution was added ethanolamine (0.16 ml) and triethylamine (of 0.64 ml) in THF (50 ml). Stirring was continued for additional 10 minutes in a bath with ice and then stirred at room temperature overnight. Then the reaction mixture was filtered, and the solvent was removed under reduced pressure. The residue was dissolved in a mixture of acetic acid (24 ml) and water (10 ml)and the solution was heated to 70C for 1 hour. After cooling to room temperature the mixture was extracted with chloroform (225 m is), and the solvent was removed under reduced pressure. The residue was dissolved in a mixture of isopropanol (50 ml) and dichloromethane (18 ml). Thereto was added potassium carbonate (5.0 g) in water (10 ml)and the resulting mixture was heated to 35-40Statem was added dropwise over 45 minutes a solution of dimethylsulfate (1 ml) in 10 ml of isopropanol. After an additional 90 minutes, the mixture was extracted with chloroform (350 ml), and the solvent was removed under reduced pressure, thus obtaining 1,10 grams of 1-hexadecyl-2-(4'-carboxymethyl)butyl-glycero-3-phosphocholine (yield 92%).

Getting 1-hexadecyl-2-(5'-carboxy)butyl-glycero-3-phosphocholine

1-Hexadecyl-2-(4'-carboxymethyl)butyl-glycero-3-phosphocholine was dissolved in methanol (25 ml). Sodium hydroxide (1.0 g)dissolved in 90% methanol (20 ml)was added to a methanol solution, and the reaction mixture was stirred at room temperature for 5 hours. The pH value of the reaction mixture was brought to 4 by adding sodium dihydrophosphate. Was added water (50 ml) and chloroform (50 ml), the organic phase was collected, and the solvent was removed under reduced pressure. The residue was dissolved in chloroform, dried over anhydrous Na2SO4, was filtered, and the solvent was removed under reduced pressure. Received 1-hexadecyl-2-(4'-carboxy)butyl-glycero-3-phosphocholine (0,71 g) (yield 66%).

Whereas, some sign of the invention, which, for clarity, described in the context of separate embodiments, may be offered in combination in a single embodiment. Conversely, various features of the invention, which, for brevity, described in the context of a variant of implementation, can also be offered separately or in any suitable subcombination.

Although the invention is described in connection with specific variants of its implementation, it is obvious that many changes, modifications, and variations will be obvious to specialists in this field. Accordingly it is intended to include all such changes, modifications and variants to include in the entity and the total amount of the attached claims. All publications, patents and patent applications mentioned in this description are listed here in their entirety by reference in the description, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to introduce here as a reference. In addition, citation or specifying any references in this application should not be construed as an admission that such reference is available as prior art to the present invention.

1. The method of obtaining compounds with glycerin main chain and at the one ore containing oxidized group residue, attached to glycerol main chain through an ether bond, and the compound has the General formula II:

where A1represents CH2;
And2represents CH2;
R1represents alkyl having 1-30 carbon atoms;
R2represents a

where
X is an alkyl chain having 1-24 carbon atoms;
Y is selected from the group consisting of hydrogen and alkyl; and Z represents-C(=O)IT; and
R3represents a phosphorus-containing part selected from the group consisting of phosphoric acid residue, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine and fosfoglitserin,
moreover, the method includes
receiving the first connection with glycerin main chain and at least one free hydroxyl group, with the specified first compound has a General formula I:

in which the Oh A 1and R1are as defined for General formula II, and R3selected from the group consisting of hydrogen, specified involved in phosphorus-containing part and reactive fosforsoderzhashchie group, which can turn into a specified phosphorus-containing part;
receiving the second connection C4-30hydrocarbon having at least one unsaturated bond and a reactive group capable of forming simple radio communication with the specified free hydroxyl group of the specified first connection;
the interaction of the specified reactive group of the second compound with a free hydroxyl group of the first connection, to obtain thereby a third connection, which is a product of the merger of unsaturated C4-30hydrocarbon to the specified glycerin main chain through an ether linkage at the sn-2-position;
the allocation of a third compound, to obtain thereby a purified third compound;
the interaction of the purified third compound with an oxidizing agent capable of oxidizing the specified unsaturated bond with the formation of the specified Z, to obtain a fourth connection,
the selection of the specified fourth compounds for the production of purified compounds of General formula II,
and specified the fourth link which has glycerin main chain and containing oxidized group Z residue, attached to the specified glycerin main chain through an ether linkage;
where, when R3specified General formula I represents a specified phosphorus-containing portion, the fourth compound is a compound of formula II,
where, when R3specified General formula I represents hydrogen, a fourth connection differs from the compounds of formula II the fact that the group R3represents a hydrogen atom or a protective group, and the interaction of these first and second connection instead of R3in the compound of the formula I are introducing a protective group which is removed prior to extraction of the specified third connection, before the interaction of the third compound with an oxidizing agent, before allocating the specified fourth connection or after allocation of the fourth connection, and additionally spend the interaction of the specified fourth connection or purified fourth compound with a reactive phosphorus-containing compound having one reactive group capable of interacting with a free hydroxyl group at position R3and another reactive group capable of being converted into the specified phosphorus-containing part, and then the resulting product is converted into a compound of formula II,
and where, when R3in connected and indicated formula I is a reactive phosphorus-containing group, a fourth connection is different from the compounds of formula II have instead of R3specified reactive fosforsoderzhashchie group, which in turn indicated the phosphate part of obtaining the compounds of formula II,
moreover, the method eliminates the use of column chromatography.

2. The method according to claim 1, where the selection is specified third connection includes:
the collection specified third connection;
obtaining a solution of a specified third compound in the solvent, whereby the solvent is chosen so that the third compound was soluble in it, and impurities formed during the reaction, were insoluble therein, to thereby obtain a mixture, which includes the above solution of the third connection in the specified solvent and insoluble impurities;
remove insoluble impurities; and
deletes the specified solvent, obtaining thereby a purified third compound.

3. The method according to claim 1, where the specified oxidizing agent selected from the group consisting of formic acid, hydrogen peroxide, periodate, perchlorate, bismuthate, permanganate, chlorite, ozone, silver oxide, osmium tetroxide, and any combination thereof.

4. The method according to claim 1, where the interaction of the purified third compound with an oxidizing agent includes:
the transformation of the purified third compound in connection with glitserinovoi the main chain and containing aldehyde residue, attached to the specified glycerin main chain through an ether linkage; and
oxidation of the compounds having glycerin main chain and containing aldehyde residue, attached to the specified glycerin main chain, to obtain the compounds having glycerin main chain and containing carboxylic acid residue attached to the specified glycerin main chain through an ether linkage.

5. The method according to claim 1, where the interaction of the purified third compound with the specified oxidizing agent includes:
the transformation of the purified third compound in connection with glycerin main chain and containing epoxide residue attached to the specified glycerin main chain through an ether linkage; and
oxidation of the compounds having glycerin main chain and containing epoxide residue attached to the specified glycerin main chain, to obtain the compounds having glycerin main chain and containing carboxylic acid residue, attached to the said main chain through an ether linkage.

6. The method according to claim 1, where the specified R3the first connection is a hydrogen.

7. The method according to claim 6, where the protective group is trityl.

8. The method according to claim 5, where R3specified the first connection of the program represents hydrogen.

9. The method of claim 8, where the protective group is trityl.

10. The method according to claim 9, further comprising before the interaction of the third compound and oxidizing agent:
replace the specified trityl protecting group selected from the group consisting of acetate, pivaloate or benzoate.

11. The method according to claim 1, where the reactive fosforsoderzhashchie compound is a phosphorus oxychloride (POCl3).

12. The method of obtaining compounds with glycerin main chain and at least one containing oxidized group residue attached to glycerol main chain through an ether bond, and the compound has the General formula II:

where A1represents CH2;
And2represents CH2;
R1represents alkyl having 1-30 carbon atoms;
R2represents a

where X is an alkyl chain having 1-24 carbon atoms;
Y is selected from the group consisting of hydrogen and alkyl; and
Z represents-C(=O)IT; and
R3represents a phosphorus-containing part selected from the group consisting of phosphoric acid residue, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phospho what ylmethanol, phosphorilation, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine and fosfoglitserin,
moreover, the method includes
receiving the first connection with glycerin main chain and at least one free hydroxyl group, with the specified first compound has a General formula I:

where A1and R1are as defined for General formula II, and R3selected from the group consisting of hydrogen, specified involved in phosphorus-containing part and reactive fosforsoderzhashchie group, which can turn into a specified phosphorus-containing part;
obtaining a fifth connection having at least one oxidized group, which represents an ester group and one reactive group capable to form an ether linkage with the specified free hydroxyl group of the first connection;
the interaction of the specified reactive group of the fifth connection with the free hydroxyl group of the first connection with obtaining a reaction mixture containing the sixth connection, which is the first product of accession containing ester group of the residue to the specified glycerin main chain through an ether linkage at the sn-2-position; and
the allocation specified in the sixth connection with obtaining a dedicated sixth connection and/or selection of the compounds having General formula II, receiving the selected compound having the General formula II;
where, when R3in connection specified General formula I represents a phosphorus-containing part, specified the sixth compound different from the compounds of formula II that instead Z is ester group, which amyraut with obtaining the compounds of Formula II;
where, when R3specified General formula I represents hydrogen, sixth compound different from the compounds of formula II that instead Z is ester group and R3represents a hydrogen atom or a protective group, and the interaction of these the first and the fifth connection instead of R3in the compound of the formula I are introducing a protective group which is removed prior to selection or after selection of the sixth connection, then additionally spend interaction sixth connection or a dedicated sixth connection with a reactive phosphorus-containing compound having one reactive group capable of interacting with a free hydroxyl group at position R3the sixth connection, and another reactive group capable of developing the specified phosphorus-containing part, obtaining compounds having a reactive phosphorous-containing group attached to the glycerol main chain;
the transformation specified reactive fosforsoderzhashchie group specified in phosphorus-containing part,
saponification of the specified ester group to obtain the compounds of formula II, and
where, when R3specified General formula I represents a specified reactive phosphorus-containing group, specified the sixth compound different from the compounds of formula II that instead Z is ester group, and R3is a specified reactive phosphorus-containing group, with the compound of formula II is obtained by transforming the specified reactive fosforsoderzhashchie group specified in phosphorus-containing portion and a saponification specified ester groups.

13. The method according to item 12, where the interaction of the specified first connection and the fifth connection is carried out in the presence of a base.

14. The method according to item 13, where the specified base selected from the group consisting of sodium hydride, sociallyengaged, sodium amide, sodium hydroxide and any mixture thereof.

15. The method according to item 12, where the specified R3the specified first connection represents hydrogen.

16. The method according to item 12, where the specified reactive fosforsoderzhashchie with whom the Union represents phosphorus oxychloride (POCl 3).

17. Route of administration involved in phosphorus-containing part in connection with glycerin main chain with at least one free hydroxyl group and having an oxidized portion attached via ether bond, as defined in claims 1 and 12, or unsaturated C4-30hydrocarbon attached through ether linkage as defined in claim 1, including:
obtaining connection with glycerin main chain with at least one free hydroxyl group and containing oxidized portion attached to the specified glycerin main chain through an ether linkage, as defined in claims 1 and 12 or the unsaturated C4-30hydrocarbon attached to glycerol main chain through an ether linkage as defined in claim 1;
the interaction indicated a free hydroxyl group of the compounds with POCl3to obtain compounds having the oxidized part or unsaturated C4-30hydrocarbon and reactive phosphorous-containing group, as defined in claims 1 and 12; and
the transformation specified reactive fosforsoderzhashchie group specified in phosphorus-containing part.

18. The method according to 17, where the specified connection with glycerin main chain, has an unsaturated C4-30hydrocarbon attached through a simple EF the RNA the connection.

19. The method of obtaining compounds with glycerin main chain and at least one containing oxidized group residue attached to glycerol main chain through an ether bond, and the compound has the General formula II:

in which
A1represents CH2;
And2represents CH2;
R1represents alkyl having 1-30 carbon atoms;
R2represents a

where X is an alkyl chain having 1-24 carbon atoms;
Y is selected from the group consisting of hydrogen and alkyl; and
Z represents-C(=O)IT; and
R3represents hydrogen,
moreover, the method includes
receiving the first connection with glycerin main chain and at least one free hydroxyl group, with the specified first compound has a General formula I:

where A1and R1are as defined above, and R3is triphenylmethyl;
obtaining a second connection, which represents a C4-30hydrocarbon having unsaturated bond and a reactive group containing a halide;
the interaction of the specified reactive group of the specified second connection with the decree which authorized the free hydroxyl group of the specified first connection to receive a third connection, which is a product of the merger of unsaturated C4-30hydrocarbon to the specified glycerin main chain through an ether linkage at the sn-2-position;
purification of the specified third connection removing insoluble impurities by filtration of the solution indicated the third connection;
oxidation of the specified bonds of specified third connection through:
(i) sequential interaction of hydrogen peroxide in formic acid, periodate and chlorite;
(ii) serial impact chloroperbenzoic acid, periodate and chlorite; or
(iii) exposure to a mixture of periodate and permanganate,
to obtain, thereby, the specified connection has indicated General formula II; and
purification of the compounds having General formula II, by recrystallization,
moreover, the method eliminates the use of column chromatography.

20. The method of obtaining compounds with glycerin main chain and at least one containing oxidized group residue attached to glycerol main chain through an ether bond, and the compound has the General formula II:

where A1represents CH2;
And2represents CH2;
R1represents alkyl having 1-30 atoms is of Pereda;
R2represents a

where X is an alkyl chain having 1-24 carbon atoms;
Y is selected from the group consisting of hydrogen and alkyl; and Z represents-C(=O)IT; and
R3represents a phosphorus-containing part selected from the group consisting of phosphoric acid residue, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine and fosfoglitserin,
moreover, the method includes
receiving the first connection with glycerin main chain and at least one free hydroxyl group, with the specified first compound has a General formula I:

where A1and R1are as defined for formula II, and R3selected from the group consisting of hydrogen, specified involved in phosphorus-containing part and reactive fosforsoderzhashchie group, which can turn into a specified phosphorus-containing part;
receiving the second connection etc is stableuser a C 4-30hydrocarbon having at least one unsaturated bond and a reactive group capable of forming simple radio communication with the specified free hydroxyl group of the specified first connection;
the interaction of the specified reactive group of the specified second connection with the specified free hydroxyl group of the specified first connection to receive a third connection, which is a product of the merger of unsaturated C4-30hydrocarbon to the specified glycerin main chain through an ether linkage at the sn-2-position;
the selection of the specified third connection with obtaining purified third compound;
oxidation of the specified bonds through:
(i) serial impact chloroperbenzoic acid, periodate and chlorite; or
(ii) exposure to a mixture of periodate and permanganate, to obtain thereby the fourth connection with glycerin main chain and containing oxidized group residue attached to glycerol main chain through an ether linkage; and
the selection of the specified fourth connection with obtaining purified fourth compound, obtaining a compound of General formula II, where
(a) R3in the compound of General formula I represents a phosphorus-containing part, and the fourth the second compound is a compound of formula (II); or
(b) R3in the compound of General formula I represents a specified reactive phosphorus-containing group, and a fourth connection differs from the compounds of formula (II) presence of reactive fosforsoderzhashchie group instead of R3and the method further includes the transformation of the specified reactive fosforsoderzhashchie group specified in phosphorus-containing part before or after allocation of the fourth connection with obtaining the compounds of formula (II); or
(c) R3in the compound of General formula I represents hydrogen, and the fourth compound different from the compounds of formula (II) those that the group R3represents a hydrogen atom or a protective group, and the interaction of these first and second connection instead of R3in the compound of the formula I are introducing a protective group which is removed prior to extraction of the specified third connection, before oxidation of the specified bonds, before or after allocation of the fourth connection, and the method further includes the interaction of the fourth connection or purified fourth compound with a reactive phosphorus-containing compound having a reactive group capable of interacting with a free hydroxyl group at position R3and another reactio nesposobny group, able to turn into a specified phosphorus-containing part, and then the resulting product is converted into a compound of formula II, or
(d) R3in the compound of General formula I represents hydrogen and the fourth compound is a compound of formula II and before the interaction of the first and second connection introducing a protective group into the compound of General formula I instead of R3and the method further includes removing the specified protective group for the selection of the third connection or before oxidation specified unsaturated communication and interaction specified third connection or purified third compound with a reactive phosphorus-containing compound having a reactive group capable of interacting with the specified free hydroxyl group at position R3and another reactive group capable of being converted into the specified phosphorus-containing part,
moreover, the method eliminates the use of column chromatography.

21. The method according to claim 20, where the reactive fosforsoderzhashchie compound is a phosphorus oxychloride (POCl3).

22. The method of obtaining compounds with glycerin main chain and at least one containing oxidized group residue attached to glycerol main chain through ether tie is, moreover, the compound has a General formula II:

where A1represents CH2;
And2represents CH2;
R1represents alkyl having 1-30 carbon atoms;
R2represents a

where X is an alkyl chain having 1-24 carbon atoms;
Y is selected from the group consisting of hydrogen and alkyl; and
Z represents-C(=O)IT; and
R3represents a phosphorus-containing part selected from the group consisting of phosphoric acid residue, phosphorylcholine, phosphorylethanolamine, phosphorylcholine, phosphorylethanolamine, fosfaurilirutisa, ethylphosphonic, phosphorylmethyl, phosphorylates, fosforilirovaniya, phosphorylmethyl, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], postinst-4-phosphate, postinst-4,5-biphosphate, pyrophosphate, phosphatidylethanolamine, dinitrobenzenamine and fosfoglitserin,
moreover, the method includes
receiving the first connection with glycerin main chain and at least one free hydroxyl group, with the specified first compound has a General formula I:

where A1and R1are as defined for formula II, and R 3selected from the group consisting of hydrogen, specified involved in phosphorus-containing part and reactive fosforsoderzhashchie group, which can turn into a phosphorus-containing part;
obtaining a second connection, which represents a C4-30hydrocarbon having at least one unsaturated bond and a reactive group capable of forming simple radio communication with the specified free hydroxyl group of the specified first connection;
the interaction of the specified reactive group of the specified second connection with the specified free hydroxyl group of the specified first connection to receive a third connection, which is a product of the merger of unsaturated C4-30hydrocarbon to the specified glycerin main chain through an ether linkage at the sn-2-position;
the selection of the specified third connection with obtaining purified third compound;
oxidation specified unsaturated due to the formation of the specified Z, to obtain a fourth connection with glycerin main chain and containing oxidized group residue attached to glycerol main chain through an ether linkage; and
the selection of the specified fourth connection with obtaining purified fourth compound, and thus, Obedinenie General formula II, where
(a) R3in the compound of General formula I represents a specified phosphorus-containing part, and the fourth compound is a compound of formula (II); or
(b) R3in the compound of General formula I represents a reactive phosphorous-containing group, and a fourth connection differs from the compounds of formula (II) presence of a specified reactive fosforsoderzhashchie group instead of R3and the method further includes the transformation of the specified reactive fosforsoderzhashchie group specified in phosphorus-containing part before or after allocation of the fourth connection with obtaining the compounds of formula (II); or
(c) R3in the compound of General formula I represents hydrogen, and the fourth compound different from the compounds of formulas (II) the fact that R3represents a hydrogen atom or a protective group, and the interaction of these first and second connection instead of R3in the compound of the formula I are introducing a protective group which is removed prior to extraction of the specified third connection, before oxidation of unsaturated communication, before or after allocation of the fourth connection, and the method further includes the interaction of the fourth connection or purified fourth compound with POCl3then the product p is euromalt in the compound of formula II, or
(d) R3in the compound of General formula I represents hydrogen and the fourth compound is a compound of formula II, and introducing a protective group into the compound of General formula I instead of R3before the interaction of the specified first and the second connection, and the method further includes removing the specified protective group with the release of the third connection or before oxidation specified unsaturated communication and interaction specified third connection or purified third compound with POCl3,
moreover, the method eliminates the use of column chromatography.



 

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SUBSTANCE: invention concerns the new synthetic oxidised lipids and ways of application of the oxidised lipids for treatment and inflammation prevention, associated with endogenous oxidised lipid.

EFFECT: rising of treatment efficiency.

54 cl, 25 dwg, 15 ex

FIELD: biotechnology.

SUBSTANCE: claimed method includes interaction of mixtures of natural phosphatides or components thereof, for instance, soybean or egg lecithin or animal phospholipids, or synthetic phosphatides, by reaction thereof with D phospholipase having transphosphatidylase activity in water medium.

EFFECT: simplified method for isolation of pure phosphatides of improved purity with increased yield.

28 cl, 13 ex

The invention relates to the field of medicine and for the application of hydrogenated soy phosphatidylcholine (Phospholipon-N) as an active ingredient, providing a surface-active properties of drugs for substitution of the surfactant therapy
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The invention relates to organic chemistry, namely the chemistry of fullerenes

The invention relates to new derivatives of phospholipids, namely alkylphosphates or ALCOHOLATES, kalinovy residue which is part of a heterocyclic ring, the method of obtaining a class of these compounds, as well as to medicines and the way to obtain medicines that contain compounds as biologically active substances

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FIELD: biotechnology.

SUBSTANCE: claimed method includes interaction of mixtures of natural phosphatides or components thereof, for instance, soybean or egg lecithin or animal phospholipids, or synthetic phosphatides, by reaction thereof with D phospholipase having transphosphatidylase activity in water medium.

EFFECT: simplified method for isolation of pure phosphatides of improved purity with increased yield.

28 cl, 13 ex

FIELD: medicine.

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54 cl, 25 dwg, 15 ex

FIELD: chemistry.

SUBSTANCE: in the proposed methods, the hydroxyl group of the formula I compound is substituted with the corresponding residue so that use of column chromatography is avoided, , where A1 is CH2, R1 is alkyl, R3 is a residue of phosphoric acid, phosphorylcholine, phosphoryl ethanolamine, phosphorylserine, phosphoryl cardiolipin, phosphoryl inosite, ethylphosphocholine, phosphoryl methanol, phosphoryl ethanol, phosphoryl propanol, phosphoryl butanol, phosphoryl ethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propyleneglycol)], phosphoinosite-4-phosphate, phosphoinosite-4,5-biphosphonate, pyrophosphate, phosphoethanolaminediethylene-triaminepentaacetate, dinitrophenyl phosphoethanolamine, phosphogycerine or a reactive phosphorus-containing group which can convert to the said residues, or hydrogen.

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22 cl, 5 ex

FIELD: medicine, pharmaceutics.

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12 cl, 3 dwg, 5 ex, 5 tbl

FIELD: chemistry.

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8 cl, 5 tbl, 1 ex

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39 cl, 11 tbl, 15 ex, 25 dwg

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