Phosphonate derivatives

FIELD: organophosphorus compounds, medicine.

SUBSTANCE: invention relates to new biologically active phosphonate compounds. Invention describes phosphonate compound of the formula:

wherein R₁ and R'₁ represent independently hydrogen atom (-H) substituted possibly with -O-(C₁-C₂₄)-alkyl, -O-(C₁-C₂₄)-alkenyl, -O-(C₁-C₂₄)-acyl, -S-(C₁-C₂₄)-alkyl, -S-(C₁-C₂₄)-alkenyl or -S-(C₁-C₂₄)-acyl wherein at least one among R and R'₁ doesn't represent -H and wherein indicated alkenyl or acyl comprise from 1 to 6 double bonds; R₂ and R'₂ represent independently -H substituted possibly with -O-(C₁-C₇)-alkyl, -O-(C₁-C₇)-alkenyl, -S-(C₁-C₇)-alkyl, -S-(C₁-C₇)-alkenyl, -O-(C₁-C₇)-acyl, -S-(C₁-C₇)-acyl, -N-(C₁-C₇)-acyl, -NH-(C₁-C₇)-alkyl, -N-((C₁-C₇)alkyl)₂, oxo-group, halogen atom, -NH₂, -OH or -SH; R₃ represents phosphonate derivative of nucleoside or biphosphonate; X represents compound of the formula:

L represents a valence bond or a bifunctional binding molecule of the formula: -J-(CR₂)_t-G- wherein t is a whole number from 1 to 24; J and G represent independently -O-, -S-, -C(O)O- or -NH-; R represents -H, unsubstituted or substituted alkyl or alkenyl; m means a whole number from 0 to 6; n = 0 or 1. Also, invention describes pharmaceutical compositions comprising phosphonate compounds, method for treatment of osteoporosis in mammal, method for increasing mineral osseous density, method for prophylaxis of apoptosis of osteoblasts and osteocytes in mammal, method for treatment of viral infection in mammal, method for treatment of growing neoplasm in mammal and method for proliferation of cells. Invention provides preparing new compounds eliciting useful biological properties.

EFFECT: valuable medicinal properties of phosphonate compounds.

17 cl, 2 dwg, 7 tbl, 21 ex

The SCOPE of the INVENTION

The present invention relates to novel phosphonate compounds containing compositions, methods for their preparation and to their use for the treatment of several medical disorders, such as osteoporosis and other disorders of bone metabolism, cancer, viral infections and the like.

PRIOR art

It has long been known that the phosphonate compounds are useful in many therapeutic aspects. A special class of therapeutically useful phosphonate compounds are bisphosphonates, i.e. analogues of pyrophosphates, in which the Central oxygen atom pyrophosphate connection is replaced by a carbon atom. This Central carbon atom can be attached to different groups-deputies, forming derivative compounds, phosphonates, which have various degrees of pharmacological effectiveness. These derivatives have the General structure

where R_aand R_bcan be independently selected from hydroxyl, amino, sulfhydryl, halogen or multiple alkyl or aryl groups or combinations of such groups, which may be optionally substituted. Examples include etidronate, where R_arepresents CH₃and R_bis a HE; clodronate, dichlorodibenzofuran acid (Cl₂MDP), where R and R_bare Cl; pamidronate, 3-amino-1-hydroxypropyltrimonium acid, where R_arepresents acylamino and R_brepresents a hydroxyl; alendronate, 4-amino-1-hydroxyethylidenediphosphonic acid, where R_arepresents propylamino and R_brepresents a hydroxyl; olpadronate, 3-dimethylamino-1-hydroxypropyltrimonium acid, where R_ais dimethylaminoethyl and R_brepresents a hydroxyl; amino-olpadronate (IG-9402), 3-(N,N-dimethylamino)-1-aminopropylphosphonic, where R_ais dimethylaminoethyl and R_brepresents NH₂.

The bifosfonatami and their substituted derivatives inherent property of inhibiting bone resorption in vivo. Bisphosphonates also inhibit apoptosis (programmed cell death) of bone-forming cells. Therefore, indications for their use include the treatment and prevention of osteoporosis, treatment of Paget's disease, cancer with bone metastases, hyperparathyroidism, rheumatoid arthritis, algodystrophy, Sterno-costal-Kuching hyperostosis, Gaucher disease, diseases of reliable community and some noskeletnih disorders (Papapoulos, S.. in Osteoporosis. R. Marcus, D. Feldman and J. Kelsay, eds., Academic Press, San Diego, 1996, p. 1210, Table 1).

Although bisphosphonates have therapeutically useful properties, ka is orally administered agents they have pharmacological disadvantages. One of the drawbacks is the low availability of oral introduction: gastrointestinal absorbed from 0.7 to 5% of the oral input dose. In addition, the absorption of oral decreases when taking with food. Then, it is known that some currently available bisphosphonates, such as FOSAMAX^TM(Merck; alendronate sodium), SKELID^TM(Sanofi, tiludronate) and ACTONE^TM(Procter and Gamble, risedronate) have local toxicity causing irritation and ulceration of the esophagus. Other bifosfonatami, such as amino-olpadronate, no antiresorptive effects (Van Beek, E. et al., J Bone Miner Res 11(10): 1492-1497 (1996)), however, they inhibit the apoptosis of osteocytes and is able to stimulate the formation of spongy bone structure (Plotkin, L. et al., J Clin Invest 104(10): 1363-1374 (1999) and U.S. patent No. 5885973). Therefore, it would be useful to develop chemically modified derivatives of bifosfonatami, which would have maintained or increased pharmacological activity of the parental compounds, at the same time eliminating or reducing their unwanted side effects.

In addition bifosfonatami, it is known that monophosphate also provide therapeutic benefit. One class of therapeutically useful monophosphates are antiviral nucleotidase, such as, for example, cidofovir, siklietotni, adefovir, tenof the VIR and the like, and 5'-phosphonates and methylenephosphonate of azidothymidine, ganciclovir, acyclovir, and the like. In compounds of this type 5'-hydroxyl of a sugar group or its equivalent in acyclic nucleosides, which do not contain the full sugar groups (ganciclovir, penciclovir, accpower), substituted communication phosphorus-carbon. If methylenephosphonate methylene group replaces the 5'-hydroxyl or its equivalent, and its carbon atom, in turn, covalently linked to the phosphonate. Below are the different patterns of AZT (azidothymidine), including compounds that are supposed to be used in the practical implementation of the present invention. Actually AZT is shown to the left. Connection And represents AZT-monophosphate, which has a regular fictitiou the link between sugar and phosphate. Unlike compounds And compounds (AZT-5'-phosphonate) and (AZT-5'-methylenephosphonate) missing 5'-hydroxyl-3'-azido-2',3'-dideoxyribose, he replaced or communication phosphorus-carbon (AZT-phosphonate), or a methylene attached communication phosphorus-carbon (AZT-methylenephosphonate). Compounds b and C are examples of compounds useful in the practical implementation of the present invention.

Compounds of this type can be active as anti-proliferative or anti-viral nucleotides. When it is Atochem metabolism occur two additional phosphorylation with the formation of the nucleotide-phosphonate-diphosphate, which is the equivalent of nucleoside-triphosphates. Antiviral nucleotide-phosphonate-diphosphate are selective inhibitors of viral RNA or DNA polymerase or reverse transcriptase inhibitors. It should be noted that their inhibitory effect on viral polymerase is much more than the degree of inhibition of their DNA-polymerases α, β and γ mammalian cell or RNA polymerase mammals. On the contrary, antiproliferative nucleotide-phosphonate-diphosphate inhibit DNA and RNA polymerases in cancer cells and may show a much lower selectivity in comparison with normal cellular DNA and RNA polymerases. As the nucleotide-phosphonates are absorbed poorly from the gastrointestinal tract, they often require parenteral administration (e.g., cidofovir). Moreover, negatively charged phosphonate group can prevent the penetration into the cell, resulting in reduced antiviral or antiproliferative activity. It was unexpectedly found that the compounds according to the invention can overcome the shortcomings of this class of agents.

Known pharmacologically active agents representing antiviral phosphonates; the following U.S. patents describe other solutions for nucleotide-phosphonate analogues: 5672697 (nucleoside-5'-methylenephosphonate), 5922695 (against the viral phosphonomethoxy-nucleotide analogs), 5977089 (antiviral, phosphonomethoxy-nucleotide analogs), 6043230 (antiviral, phosphonomethoxy-nucleotide analogs), 6069249. Previously disclosed the preparation and use of alkylglycerols, covalently linked c not containing phosphonate drugs, which are amino, carboxyl, hydroxyl or sulfhydryl functional groups. These prodrugs may contain a linking group or one or two additional phosphate ester groups between the medicinal product and allylglycidylether (U.S. patent No. 5411947 and U.S. patent application serial number 08/487081). Known partial esters chloromethylphosphonic acid (U.S. patent No. 5376649) and were reported dianhydride clodronate (Ahlmark, et al, J Med Chem 42: 1473-1476 (1999)). However, found that the partial esters are unable to release the active biphosphonate chemical or biochemical transformation (Niemi, R. et al., J Chrom In 701:97-102 (1997)). Also described prodrugs containing alkylphenolate remains associated with antiviral nucleosides (U.S. patent No. 5223263) or phosphono-carboxylates (U.S. patent No. 5463092).

Therefore, there is a constant need for less toxic and more effective pharmaceutical agents to treat a variety of disorders, such as disorders caused by a viral infection and inappropriate about what operacia cells, for example cancer. Thus, the present invention is to develop chemically modified phosphonate derivatives pharmacologically active agents such as antiviral and anticancer pharmaceutical agents. These modified derivatives strengthen the effectiveness of the parental connection, reducing harmful side effects when administered to a subject in need of it.

SUMMARY of the INVENTION

According to the invention proposed analogues phosphonate compounds. Connection phosphonates considered for use in accordance with the invention, include those phosphonate compounds that reduce bone resorption or inhibit apoptosis of osteoblasts or osteocytes, as well as those compounds that improve biological activity, selectivity or bioavailability nucleotide-phosphonate analogs, which are useful for the treatment of cancer, various viral infections and the like. Compounds according to the invention contain phosphonates, covalently linked (directly or indirectly, i.e. via a linking molecule) with a substituted or unsubstituted alkylglycerol, alkyldiphenyl, alkylation or related group. In accordance with another aspect of the present invention proposed pharmaceutical preparations containing an the logs described here phosphonate compounds.

In accordance with another aspect of the present invention offers a number of treatment methods, for example methods of treating or preventing bone resorption in a mammal, methods of enhancing bone formation by prevention of apoptosis of osteoblasts or osteocytes, ways to increase mass and bone strength, methods of treatment of viral infections, treatment of disorders caused by inappropriate cell proliferation, such as cancer, and the like.

BRIEF DESCRIPTION of DRAWINGS

Figure 1 summarizes the effect of the compounds according to the invention is 1-O-hexadecyloxypropyl-alendronate on caused by dexamethasone apoptosis of osteocytes MLO-Y4. Columns represent the average value of ± standard deviation of three independent measurements. Light columns indicate the absence of dexamethasone, and the dark columns indicate the presence of 10^-4M dexamethasone.

Figure 2 summarizes the effect of the compounds according to the invention is 1-O-hexadecyloxypropyl-alendronate caused by dexamethasone apoptosis of cells of the cranial vault. Columns represent the average value of ± standard deviation of three independent measurements. Grey columns indicate the absence of dexamethasone, and the black columns indicate the presence of 10^-4M dexamethasone.

DETAILED description of the INVENTION

Phosphonate the e compounds according to the invention have the following structure:

where R₁and R₁' independently represent-H, possibly substituted-O(C₁-C₂₄)alkyl, -O(C₁-C₂₄)alkenyl, -O(C₁-C₂₄)acyl, -S(C₁-C₂₄)alkyl, -S(C₁-C₂₄)alkenyl or-S(C₁-C₂₄)acyl, and at least one of R₁and R₁' is not-h, and moreover, these alkeneamine or acyl group may have from 1 to 6 double bonds;

R₂and R₂' independently represent-H, possibly substituted-O(C₁-C₇)alkyl, -O(C₁-C₇)alkenyl, -S(C₁-C₇)alkyl, -S(C₁-C₇)alkenyl, -O(C₁-C₇)acyl, -S(C₁-C₇)acyl, -N(C₁-C₇)acyl, -NH(C₁-C₇)alkyl, -N((C₁-C₇)alkyl)₂, oxo, halogen, -NH₂HE or SH;

R₃is a pharmaceutically active phosphonate, biphosphonate or phosphonate derivative pharmacologically active compounds that are associated with the functional group of a possible link L or with the available oxygen atom at C^α;

X, if present, is a

L represents a valence bond or a bifunctional linking molecule of the formula-J-(CR₂)_t-G-, where t is the number the t 1 to 24, J and G independently represent-O-, -S-, -C(O)O - or-NH-, a R represents-H, a substituted or unsubstituted alkyl or alkenyl;

m is an integer from 0 to 6, and

n is 0 or 1.

In preferred embodiments m is 0, 1 or 2. In these preferred embodiments, R₂and R₂' preferably represent-H, and prodrugs are then tandilia, propantheline or butadiene derived medicinal phosphonate. A preferred class tenderfoot has the following structure:

where R₁, R₁', R₃L and n are such as defined above.

Preferred prandially class has the following structure:

where m=1, a R₁, R₁', R₃L and n are such as defined above in General formula.

Preferred glycerol class has the following structure:

where m=1, R₂=H, R₂'=OH and R₂and R₂' With^αboth represent-N. Glycerin is an optically active molecule. When using the legend stereospecific numbering for glycerine position sn-3 is the position that fosfauriliruetsa glycerokinase. In the compounds according to the invention, having a glycerol residue, ruppirovka -(L) _n-R₃can be attached or sn-3, or sn-1 position of glycerol.

In all classes of pharmacologically active agents according to the invention R₁preferably represents alkoxygroup having the formula-O-(CH₂)_t-CH₃where t is 0-24. More preferably t is 11-19. Most preferably t is 15 or 17.

Preferred groups R₃include bisphosphonates, which are known to be clinically useful, for example, the following connections:

Etidronate: 1-hydroxyethylidene-diphosphonic acid (EDHP);

Clodronate: dihlormetilen-diphosphonic acid (Cl₂MDP);

Tiludronate: chloro-4-phenylthiomethyl-diphosphonic acid;

Pamidronate: 3-amino-1-hydroxypropylamino-diphosphonic acid (ADP);

Alendronate: 4-amino-1-hydroxybutylidene-diphosphonic acid;

Olpadronate: 3-dimethylamino-1-hydroxypropylamino-diphosphonic acid (dimethyl-APD);

Ibandronate: 3-methylpentylamino-1-hydroxypropylamino-diphosphonic acid (VM 21.0955);

EB-1053: 3-(1-pyrrolidinyl)-1-hydroxypropylamino-diphosphonic acid;

Risedronate: 2-(3-pyridinyl)-1-hydroxyethylidene-diphosphonic acid;

Amino-olpadronate: 3-(N,N-dimethylamino-1-aminopropylene)biphosphonate (IG9402)

and the like.

R₃can also be selected from a number postnationalism nucleotides or nucleosides which can be transformed to their corresponding derivatives phosphonates), here it is also assumed to apply for the purposes of the present invention. Preferred nucleosides include those which are useful for treating disorders caused by inappropriate cell proliferation, such as 2-chloro-deoxyadenosine, 1-β-D-arabinofuranosyl-citizen (cytarabine, Ara-C), ferritin, ftordezoksiuridin (floxuridine), gemcitabine, cladribine, fludarabine, pentostatin (2'-deoxycoformycin), 6-mercaptopurine, 6-tioguanin and substituted or unsubstituted 1-β-D-arabinofuranosyl-guanine (Ara-G), 1-β-D-arabinofuranosyl-adenosine (Ara-a), 1-β-D-arabinofuranosyl-uridine (Ara-U), and the like.

Nucleosides are useful for the treatment of viral infections, can also be converted into their corresponding 5'-phosphonates for use as group R₃. Such phosphonate analogs typically contain or phosphonate (RHO₃H₂), or methylenephosphonate (-CH₂RHO₃H₂) group, which replaced the 5'-hydroxyl antiviral nucleoside. Some examples of antiviral phosphonates are derived, obtained by replacing the 5'-hydroxyl on RHO₃H₂:

3'-azido-3',5'-dideoxythymidine-5'-phosphonic acid (AZT-phosphonate)

Hakimelahi, GH; Moosavi-Movahedi, A.A.; Sadeghi, M.M.; Tsay, S-C.; Hwu, J. R. J. Med. Chem. 1995, 38:4648-4659,

3',5'-dideoxythymidine-2'-ene-5'-phosphono the traveler acid (d4T phosphonate)

ibid.,

2',3',5'-trimethoxysilyl-5'-phosphonic acid (ddC phosphonate)

Kofoed, T., Ismail, A.E.A.A.; Pedersen, E.B.; Nielsen, S. Bull.Soc.Chim. Fr. 1997, 134:59-65,

9-[3-(phosphono-methoxy)propyl]adenine (Adefovir)

Kim, C.U.; flood, B.Y.; Misco, P. F.; Bronson, J.J.; Hithcock, M. J.M.; Ghazzouli, I.; Martin, J.C. J. Med. Chem. 1990, 33:1207-1213.

Some examples of antiviral phosphonates are derived, obtained by replacing the 5'-hydroxyl-CH₂RHO₃H₂:

Ganciclovir-phosphonate

Huffman, J.H.; Sidwell, R.W.; Momson, A.G.; Coombs, J., Reist, E.J. Nucleoside Nucleotides, 1994, 13:607-613.

Acyclovir-phosphonate

ibid.,

Ganciclovir-cyclophosphate

Smee, D.F.; Reist, E.J. Antimicrob. Agents Chemother. 1996, 40:1964-1966,

3'-thia-2',3'-dideoxycytidine-5'-phosphonic acid

Kraus, J.L; Nucleotides Nucleoside, 1993, 12:157-162.

Other preferred antiviral nukeondelete that you want to apply for the implementation in practice of the present invention, receive similar of antiviral nucleosides, including ddA, ddl, ddG, L-FMAU, DXG, DAPD, L-dA, L-dl, L-(d)T, L-dC, L-dG, FTC, penciclovir and the like.

Additionally antiviral phosphonates, such as cidofovir, cyclo-zidovu the R, adefovir, tenofovir, and the like, can be used as group R₃according to the present invention.

Certain compounds of the invention possess one or more than one chiral center, for example, in the sugar groups and, thus, can exist in optically active forms. Also, if the compounds contain alkenylphenol group or unsaturated alkyl or acyl group, there is a possibility of the presence of CIS - and TRANS-isomeric forms of such compounds. Additional asymmetric carbon atoms may be present in the group-Vice, such as an alkyl group. R - and S-isomers and their mixtures, including racemic mixtures, as well as a mixture of CIS - and TRANS - isomers are contemplated by the present invention. Understood that all such isomers and mixtures thereof are included in this invention. If desired an individual stereoisomer, it can be well-known in the field methods, using stereospecific reaction with the starting materials that contain asymmetric centers and are already separated, or methods that result in a mixture of stereoisomers and their separation by known methods.

There are many phosphonate compounds which can be converted into derivatives according to the invention, to improve their pharmacological asset is ity or increase their absorption by oral administration, as, for example, compounds described in the following patents, each of which is incorporated here fully by reference: U.S. patent nos 3468935 (Etidronate), 4327039 (Pamidronate), 4705651 (Alendronate), 4870063 (derived diphosphonic acid), 4927814 (bisphosphonates), 5043437 (phosphonates of azidodideoxythymidine), 5047533 (acyclic purine phosphonate nucleotide analogs), 5142051 (N-phosphonylmethoxyethyl derivatives of pyrimidine and purine bases), 5183815 (agents acting on the bone), 5196409 (bisphosphonates), 5247085 (antiviral purine compounds), 5300671 (gem-diphosphonic acid), 5300687 (triftormetilfosfinov), 5312954 (bis - and tetrakisphosphate), 5395826 (derived guanidinate-1,1-diphosphonic acid), 5428181 (biphosphonate derivatives), 5442101 (derived methylenephosphonic acid), 5532226 (triftormetilfosfinov), 5656745 (analogues of nucleotides), 5672697 (nucleoside-5'-methylene-phosphonates), 5717095 (analogues of nucleotides), 5760013 (analogues timedelta), 5798340 (analogues of nucleotides), 5840716 (phosphonocrotonate connection), 5856314 (tizanidine nitrogen-containing heterocyclic phosphonate compounds), 5885973 (olpadronate), 5886179 (analogues of nucleotides), 5877166 (enantiomerically pure 2-aminopurin-phosphonate-nokioteca analogues), 5922695 (antiviral, phosphonomethoxy-nokioteca analogues), 5922696 (ethylene and allene phosphonate derivatives of purines), 577089 (antiviral, phosphonomethoxy-nucleotide analogs), 6043230 (antiviral, phosphonomethoxy-nucleotide analogs), 6069249 (antiviral, phosphonomethoxy-nucleotide analogs), Belgian patent No. 672205 (clodronate), European patent No. 753523 (aminosilane diphosphonic acid); an application for a European patent 186405 (genialny bisphosphonates), and the like.

Certain biphosphonate compounds have the ability to inhibit stvalentines and cholesterol levels in the blood serum of mammals, including humans. Examples of such bifosfonatami described, for example, in U.S. patent No. 5441946 and 5563128 (Pauls et al. Phosphonate derivatives of lipophilic amines), each of which is fully incorporated here by reference. Analogues of these inhibiting stvalentines compounds according to the invention and their use in the treatment of disorders of lipid metabolism in humans are included in the scope of the present invention. Bisphosphonates according to the invention can be applied orally or topically for prevention or treatment of periodontal diseases, as described in U.S. patent No. 5270365, fully included here by reference.

The term "alkyl", as used here, refers to the monovalent radical with a straight or branched chain or cyclic radical containing from 1 to 24 carbon atoms, including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, mpem-butyl, n-hexyl and the like.

How it is used, "substituted lcil" denotes alkyl groups, optionally bearing one or more than one Deputy, selected from hydroxy, alkoxy (of a lower alkyl group), mercapto (of a lower alkyl group), cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aryloxy, replaced aryloxy, halogen, trifloromethyl, cyano, nitro, acrylic, amino, amido, -C(O)H, acyl, axially, carboxyl, carbamate, sulfonyl, Sulfuryl and the like.

As used here, "alkenyl" refers to hydrocarbon groups with straight or branched chain having one or more than one double carbon-carbon bond and containing from about 2 to 24 carbon atoms, and "substituted alkenyl" refers to alkenyl groups, optionally bearing one or more than one Deputy, as described above.

As it is used herein, "aryl" refers to aromatic groups containing from 6 to 14 carbon atoms and "substituted aryl" refers to aryl groups, optionally bearing one or more than one Deputy, as described above.

As used here, "heteroaryl" refers to aromatic groups containing one or more than one heteroatom (such as N, O, S, or the like) as part of the structure of the rings and containing in the range from 3 to 14 atom is in carbon, a "substituted heteroaryl" refers to heteroaryl groups, optionally bearing one or more than one Deputy, as described above.

The term "communication", or "valence bond"refers to the relationship between atoms, consisting of an electron pair.

As used here, the term "pharmaceutically acceptable salt" refers to salts of the accession of the acids and salts of attaching the base.

As used here, the term "prodrug" refers to a derivative of pharmaceutically active compounds which have groups otsepleniya chemically or in the course of metabolism, and become a pharmaceutically active compound in the result of solvolysis or under physiological conditions in vivo.

Phosphonate analogs containing a therapeutically effective phosphonates (or phosphonate derivatives of therapeutically effective compounds), covalently linked through the hydroxyl group of 1-O-alkylglycerol, 3-O-alkylglycerol, 1-S-alkylthiophene or alkoxy-alkanols, can be more effectively absorbed in the gastrointestinal tract than the parental compound. Oral injected dose equivalent absorbed completely from the gastrointestinal tract of a mammal, and the active drug is released in vivo under the influence of endogenous enzymes. Phosphonate analogs according to the invention can also and the geta a higher degree of biological activity compared to the corresponding compounds to obtain derivatives.

Compounds of the present invention is an improvement compared to alkylphenolate prodrugs described in the prior art, as containing phosphonate group is linked directly with alkylglycerol or alkoxyalkanols group, and because the presence of phosphonate connection prevents the enzymatic conversion of free medicine. In advanced analogues may be other links between these groups. For example, bifunctional linking units having the formula-O-(CH₂)_n-C(O)O-, where n is 1-24, can associate phosphonate with a hydroxyl group alkoxyalkanols or alkylglycerol group.

The above allows the phosphonate according to the invention to achieve a greater degree of absorption when administered orally. Moreover, cellular enzymes, but not the plasma enzymes or digestive tract, will transform this conjugate in the free phosphonate. An additional advantage of the alkoxy alkanol-phosphonates is that significantly reduces or eliminates the tendency jointly introduce food to reduce or eliminate the absorption of phosphonates, resulting in achieved higher levels in the plasma and the best that patient a treatment regimen.

Compounds according to the invention can enter the ü orally in pill form, capsules, solutions, emulsions or suspensions, in the form of liquid or solid particles inhaled in the form microencapsulating particles, in the form of a spray, through the skin by means of such devices as a transdermal patch, or rectally, for example in the form of suppositories. Lipophilic proletarienne derivatives according to the invention are particularly suitable for introduction by the absorption and transdermal delivery systems and can be used in toothpaste. The introduction can also be carried out parenterally in the form of injection solutions.

Compositions can be prepared in conventional forms, for example capsules, tablets, aerosols, solutions, suspensions or together with carriers for local application. Pharmaceutical preparations containing compounds of this invention can be prepared by conventional techniques, e.g. as described in Remington''s Pharmaceutical Sciences. 1985.

Applied pharmaceutical carrier or diluent may be a conventional solid or liquid carrier. Examples of solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, Arabic gum, magnesium stearate, stearic acid or lower alkalemia cellulose ethers. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, amines, fatty acids, polyoxyethyl is n or water. The carrier or diluent may comprise any known material for sustained release, such as glycerylmonostearate or distearate, one or a mixture of wax.

If you're using a solid carrier for oral administration, the drug can be tablet or place in the form of powder or pellets into hard gelatin capsule. The amount of solid carrier will vary widely, but typically ranges from about 25 mg to about 1, If used carrier liquid, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.

Tablets are prepared by mixing the active ingredient (i.e. one or more than one compounds according to the invention with pharmaceutically inert organic or inorganic carrier, diluents and/or excipients. Examples of excipients that can be used for tablets are lactose, corn starch or its derivatives, talc, stearic acid or its salts. Examples of suitable excipients for gelatin capsules are vegetable oils, waxes, fats, semi-solid and liquid polyols. Biphosphonate prodrugs can also be produced in microencapsulating form.

For online analnogo administration of the drug may contain the compound according to the invention, dissolved or suspended in a liquid carrier, in particular in aqueous media, for use in aerosol form. The media may contain solubilizing agents, such as propylene glycol, surfactants, amplifiers suction, such as lecithin or cyclodextrin, or preservatives.

Pharmaceutical compositions for parenteral injection according to this invention contain a pharmaceutically acceptable sterile aqueous or nonaqueous liquids, dispersions, suspensions or emulsions, and sterile powders for dilution with obtaining sterile injectable solutions or dispersions just prior to use.

Suitable excipients for the preparation of solutions and syrups are water, polyols, sucrose, invert sugar, glucose and the like. Suitable excipients for the preparation of injection solutions are water, polyols, alcohols, glycerol, vegetable oils and the like.

Pharmaceutical products may additionally contain any of the many add-on components such as, for example, preservatives, soljubilizatory, stabilizers, moistening agents, emulsifiers, sweeteners, colorants, corrigentov, buffers, covering agents, antioxidants, diluents and the like.

Optional pharmaceutical composition and the finding may contain connection, corresponding to the General formula in combination with one or more than one compound exhibiting a different activity, such as an antibiotic or other pharmacologically active material. Such combinations are included in the scope of the present invention.

According to this invention methods of treating disorders in mammals associated with bone metabolism, viral infections, inappropriate cell proliferation and the like. In particular, in these ways for a person or other mammal in need of this, introducing a therapeutically effective amount of the prodrugs of this invention. Indications for such treatment include the treatment of age, postmenopausal or caused by steroids osteoporosis, Paget's disease, cancer, bone metastasis, hyperparathyroidism, rheumatoid arthritis, algodystrophy, Sterno-costoclavicular hyperostosis, Gaucher disease, diseases of reliable community, some noskeletnih disorders and diseases of the periodontium, virus immunodeficita human (HIV), influenza virus, herpes simplex virus (HSV), human herpes virus 6, cytomegalovirus (CMV), hepatitis b virus, Epstein-Barr (EBV), varicella zoster virus, lymphoma, hematological disorders such as leukemia, and the like.

In accordance with one aspect of the invention offer the s methods of preventing or treating bone loss in a mammal, especially in humans, in which human or mammal is administered a therapeutically effective amount of the compounds according to this invention. Biphosphonate prodrugs according to the invention, inhibiting bone resorption, therapeutically useful for anti-mediated osteoclastic bone resorption or bone loss in conditions when bisphosphonates, which received the prodrug, are considered to be effective. Indications for such therapy include osteoporosis, particularly in women after menopause, osteoporosis, which is accompanied by long-term steroid therapy, bone Paget's disease. Discovered that biphosphonate connection clodronate (Ostac, Boehringer-Mannheim, Mannheim, Germany) also reduces bone and visceral metastases in patients with breast cancer at high risk of distant metastases (Diel, I.J. et al. (1998) New Engl. J. Med. 339 (60, 357-363). Efficiency biphosphonate prodrugs according to the invention can be evaluated by the same methods as the effectiveness of the parental compounds. They include comparative measurement of bone mineral density of the lumbar spine, femur neck, trochanter, forearm and whole body along with measurements of vertebral fractures, spinal deformities and growth in osteoporosis, bone scan or x-ray identification of damage to the STI in metastatic tumors and the like.

According to another aspect of the present invention proposed ways to increase the mass and bone strength in a mammal, particularly humans, which impose a stimulating bone anabolism compounds according to the invention that inhibit the apoptosis of osteoblasts and osteocytes, leading to greater speeds actually ontogenese without changing the essential functions of osteoclasts (Piotkin et al., J Clin Invest 104:1363-1374 (1999), and Van Beek et. al., Bobe J Min Res 11:1492 (1996)).

According to another aspect of the invention methods of treating disorders caused by viral infections. Indications for such treatment include sensitive viruses such as virus immunodeficita human (HIV), influenza, herpes simplex virus (HSV), human herpes virus 6, cytomegalovirus (CMV), hepatitis b and C, Epstein-Barr (EBV), varicella zoster virus, and disease caused by orthopox-viruses (for example, viruses of the major and minor variola, cowpox, smallpox, ospowiki, camel pox, monkey pox, and the like), the causative agent of hemorrhagic fever Ebola, human papilloma virus, and the like.

According to another aspect of the invention methods of treating disorders caused by inappropriate cell proliferation, such as cancer, such as melanoma, lung cancer, pancreatic cancer, racially, the colon and rectum, prostate cancer and breast cancer, leukemia, lymphoma and the like. Protivoopuxolevye compounds that can be converted into their nucleotidase for use as compounds for this invention include, but are not limited to, cytarabine (Ara-C), ferritin, ftordezoksiuridin (floxuridine), gemcitabine, cladribine, fludarabine, pentostatin (2'-deoxycoformycin), 6-mercaptopurine, 6-tioguanin, and substituted or unsubstituted Ara-adenosine (Ara-A, Ara-guanosine (Ara-G) and Ara-uridine (Ara-U). Antitumor compounds according to the invention can be used alone or in combination with other antimetabolite or other classes of anticancer drugs, such as alkaloids, topoisomerase inhibitors, alkylating agents, antitumor antibiotics, and the like.

Prodrugs according to the invention can be administered orally, parenterally, topically, rectally, and other ways in appropriate units of dosage as required.

The term "parenteral"as used here, refers to subcutaneous, intravenous, intraarterial, intramuscular injection and the injection inside the vitreous body of the eye or infusion techniques.

The term "local" includes an introduction rectally or by inhalation of the aerosol, as well as more General PU and through the skin, the mucous membranes of the mouth and nose and in toothpaste.

The term "effective amount" as applied to the phosphonate prodrugs according to the invention means the amount that will prevent or reverse the above diseases. In particular, with regard to disorders associated with bone metabolism, the effective amount is the amount that will prevent, reduce or reverse the abnormal or excessive bone resorption or bone resorption that occurs in aging, especially in menopause, women either to prevent or to counteract bone metastases, visceral metastases in breast cancer.

As for disorders associated with viral infections or inappropriate cell proliferation, such as cancer, "effective amount" is defined in accordance with the recommended dosages of the parental anti-viral or anti-cancer compounds. The selected dose will vary depending on the activity of the selected compound, route of administration, the severity of the condition being treated and the condition and prior medical history treated the patient. Knowledge in this area allow you to start taking the dosage of the compounds (compounds) at levels lower than required to achieve the desired terapeutiche is one effect, and gradually increase the dosage until the desired effect is reached. If desired, the effective daily dose can be divided into multiple doses with the aim of introducing, for example, from two to four doses per day. It should be understood, however, that a particular level of dose for an individual patient will depend on a number of factors including the body weight, General health, diet, time and route of administration, and combination with other drugs, as well as the severity of the disease that is being treated.

In General, the compounds of the present invention provide a standard dosage forms containing from 1 to 100% active ingredient. The interval at therapeutic dosage is from about 0.01 to about 1000 mg/kg/day, preferably from about 0.10 to 100 mg/kg/day when administered to patients, such as people, as a medicine. Actual dosage levels of active ingredients in the pharmaceutical compositions of this invention can be varied in order to enter the number of active compound(s), which is effective to achieve the desired therapeutic response in the individual patient.

Numerous animal experiments have shown the effectiveness of bifosfonatami in the prevention of bone loss in experimental conditions that mimic relevant clinical disorders. is based on these studies there are several small animal models to evaluate the effectiveness of bifosfonatami. These tests are also useful for measuring the relative efficiency biphosphonate prodrugs according to the invention. Assessment biphosphonates therapy usually requires determination of the mass of the femoral ash and bone mass, measured, for example, as trabecular bone volume in treated and untreated animals. Thomson, D. et al. ((1990) J. Bone and Mineral Res. 5(3):279-286) describes the application of these evaluation methods of inhibiting bone loss in immobilized rats, which were treated with aminohydroxylation-biphosphonates. Yamamoto, M. et at. (1993) Calcif Tissue Int 53:278-282, called hyperthyroidism in rats receiving bone changes similar to those in people with hyperthyroidism, and compared the group who were treated bifosfonatami and not treated, biochemically on the basis of measurements of osteocalcin and using histomorphometrical analysis, including differences in the amount of cancellous bone, and histological comparison osteogenic, osteoclastic and osteoblastic surfaces in the bone slices. Seedor, J.G. et al. ((1991) J. Bone and Mineral Res. 6(4):339-346) describes studies on the action of alendronate on antagonization of bone loss in ovariectomized rats by weight of femoral ash and by histomorphometrical the analysis of tibial trabecular bone. Analysis Schenk, including histological examination of the epiphyses of growing rats, can also be used in quality is the firmness of the screening analysis. Illustrative screening test to assess antagonizing action of the compounds according to the invention on bone resorption in laboratory rats using different techniques violated osteogenesis described in example 14.

Compounds according to the invention can be obtained in various ways, as summarised in schemes I-VI. General methods of esterification of phosphonates described below is only for illustrative purposes and should not be construed as in any way limiting this invention. Indeed, there are several ways the direct condensation of phosphonic acids with alcohols (see, for example, R.C. Larock, Comprehensive Organic Transformation, VCH, New York, 1989, p. 966, and here is there information sources). Isolation and purification of the compounds and intermediates described in the examples, you can run if you want, using any suitable methods of separation and purification, such as, for example, filtration, extraction, crystallization, column flash chromatography, thin layer chromatography, distillation or a combination of these methods. Specific illustrations of suitable separation techniques and the selection below in the examples. Other equivalent methods of division and separation, of course, can also be applied.

Scheme 1 shows the General synthesis biphosphonate prodrugs that sod is rat primary amino group, such as pamidronate or alendronate. Example 1 gives the conditions of synthesis of 1-O-hexadecyloxypropyl-alendronate (HDP-alendronate) or 1-O-hexadecyloxypropyl-pamidronate (HDP-pamidronate). In this way a mixture of dimethyl-4-phthalimidobutyl-phosphonate (1b, obtained as described in U.S. patent 5039819) and hexadecyloxypropyl-methylphosphate (2) in pyridine solution is treated with triethylamine, getting biphosphonates tetraethyl 3b, which is purified by chromatography on silica gel. Intermediate compound 2 is obtained by transesterification of diphenylphosphite, as described in (Kers, A., Kers, I., Stawinski, J., Sobkowski, M., Kraszewsku, A., Synthesis, April 1995, 427-430). So, diphenylphosphite in pyridine solution first handle hexadecyloxypropyl-1-I, then with methanol, resulting in a receive connection 2.

A significant aspect of this method is that you can apply to other long-chain alcohols instead of hexadecyloxypropyl-1-ol, receiving various compounds according to this invention. Treatment of intermediate compounds 3b-bromotrimethylsilane in acetonitrile selectively decompose methyl esters, receiving monoether 4b. Treatment of 4b with hydrazine in the mixed solvent system (20% methanol / 80% 1,4-dioxane) removes the protective phthalimide group, as shown in the diagram. The desired prodrug of alendron is and is collected by filtration and turn in tragamoneda salt by treatment with methanolic ammonia solution.

Scheme II illustrates the synthesis of analogues of bifosfonatami who does not have a primary amino group. In this case, the stages of the method are similar to those described in scheme I, except that the protection phthalimide group and the subsequent removal of the protection hydrazinolysis not needed.

Bisphosphonates with 1-amino group, such as amino-olpadronate, can be converted into analogues corresponding to the prodrugs according to the invention, using a slightly modified method, shown in scheme III.

Treatment of a mixture of compounds 2 and 3-(dimethylamino)propionitrile dry HCl followed by the addition of dimethylphosphite gives tetraethyl 3, which after demethylation by bromotrimethylsilane gives hexadecyloxypropyl-1-amino-olpadronate.

Scheme IV illustrates the synthesis of no ether phosphonate and biphosphonates analogue, in which the primary amino group attached to a lipid group parental connection.

Scheme V illustrates the General synthesis alkylglycerol or alkylphosphonium analogs of cidofovir, cyclo-cidofovir and other phosphonates. Treatment of 2,3-isopropylideneglycerol 1 NaOH in dimethylformamide and subsequent interaction with alkylarylsulfonates get alkilany ether 2. Remove isopropylidene group is the processing of acetic acid, followed by interaction with Fritillaria in pyridine gives the intermediate compound 3. Alkylation of intermediate compound 3 alkylhalogenide leads to the connection 4. Remove trailvoy group 80%aqueous acetic acid leads to O,O-diacylglyceride 5. As a result, the synthesized compounds 5 and subsequent interaction with the sodium salt of cyclo-cidofovir or other postnataldepression nucleotide get the desired phosphonate product fitting 7. Breaking the cycle cyclo-adduct is carried out by reacting with aqueous sodium hydroxide. Preferred prandially class can be synthesized by replacing the connection 5 1-O-allylprodine-3-ol in scheme V. Analogues of tenofovir and adefovir can be synthesized by replacing cyclo-cidofovir (cCDV) on these nucleotide-phosphonates in the reaction (f) scheme V. Similarly, the same method can be used to obtain other nucleotide-phosphonates according to the invention.

Reagents: a) NaH, R₁OSO₂Me, dimethylformamide (DMF); (b) 80% water. acetic acid; (C) Fritillaria, pyridine; (d) NaH, R₂-Br, DMF; f) CBr₄, triphenylphosphine; tetrahydrofuran (THF); (f) cyclo-cidofovir (salt DCMC), DMF; d) 0,5 N. NaOH

Scheme VI illustrates a General method for the synthesis of nucleotide-phosphonates according to the invention with the use of 1-O-hexadecyloxypropyl-adefovir as an example. Nucleotide-phosphonate (5 mmol) is suspended in anhydrous pyridine and add Alcock valcanale or alkylglycerol derived and 1,3-dicyclohexylcarbodiimide (DCC, 10 mmol). The mixture is heated under reflux and intensively stirred up until the condensation reaction is completed (monitored by thin layer chromatography). The mixture is then cooled and filtered. The filtrate is concentrated under reduced pressure, the residue adsorb on silica gel and purified by column flash chromatography (elution with a mixture of approximately 9:1 dichloromethane/methanol)to give the corresponding monoether phosphonate.

Now the invention will be described in more detail by addressing the following non-limiting examples.

EXAMPLE 1

Synthesis of 1-O-hexadecyloxypropyl-3-alendronate

A. Hexadecyloxypropyl(b)

Hexadecyloxypropyl was obtained by the method described in Kers, A., Kers, I., Stawinski, J., Sobkowski, M., Kraszewsku, A., Synthesis, April 1995, 427-430. To a solution of diphenylphosphine (14 g, 60 mmol) in pyridine (50 ml)maintained at 0°slowly added a solution hexadecyloxypropyl-1-ol (6.0 g, 20 mmol) in pyridine (25 ml). The mixture was stirred for 1 hour, before added anhydrous methanol (10 ml). After stirring for a further one hour, the solvent is boiled away and the residue was adsorbing on silica gel and was chromatographically using gradient elution (from hexanol and to a mixture of 20% ethyl acetate / 80% hexane), the result is what got pure compound 2 in the form of low-melting waxy solid (4.5 g, yield 60%).

¹H-NMR (CDCl₃that δ): 6.79 (d, 1H, J = 696 Hz), 4.19 (q, 2H), 3.78 (d, 3H), 3.51 (t, 3H), 3.40 (t, 2H), 1.95 (Penta, 2H), 1.25 (Shir. with, 28N), 0.88 (t, 3H).

Century Hexadecyloxypropyl-trimethyl-4-phthalimidobutyl-phosphonate(3b)

To a mixture of dimethyl-4-phthalimidobutyl-phosphonate (1b, 3,0, 7.9 mmol, obtained as described in U.S. patent 5039819) and hexadecyloxypropyl (2, 2.9 g, 9 mmol) in pyridine (50 ml) was added triethylamine (0.2 g, 2 mmol). The mixture was stirred for 5 hours at room temperature, then the solvent was removed in vacuum. The residue was absorbed on silica gel and was chromatographically (ethyl acetate) to give 3b (3.5 g, 63%) as a viscous oil.

¹H-NMR (CDCl₃): 7.84 (d, 2H), 7.72 (d, 2H), 4.45 (m, 1H), 4.27 (m, 4H), 4.15 (q, 2H), 3.68 (s, 3H), 3.84 (s, 3H), 3.71 (t, 2H), 3.51 (m, 2H), 3.38 (t, 2H), 2.04 (m, 2H), 1.94 (Penta, 2H), 1.54 (m, 2H), 1.25 (Shir. with, 28N), 0.88 (t, 3H).

³¹P-NMR (22.54 (doublet), 21.22 (Quartet)).

C. Hexadecyloxypropyl-4-phthalimidobutyl-phosphonate(4b)

Compound 3b, obtained in the previous phase (3.0 g, 4.3 mmol)was dissolved in anhydrous acetonitrile (50 ml) and cooled to 0°C. is Slowly added a solution of bromotrimethylsilane (3,9 g, 25.5 mmol) in acetonitrile (25 ml) and then the solution was further stirred for 2 hours. The mixture is then slowly poured into crushed ice. The precipitate was collected by vacuum filtration and dried in vacuum receipt is m 1.2 g 4b (yield 42%).

¹H-NMR (DMSO-d₆): 7.86 (m, 4H), 3.99 (q, 2H), 3.66-3.55 (m, 1H), 3.54 (m, 2H), 3.35 (t, 2H), 3.27 (t, 2H), 1.89-1.80 (m), 1.72 (Penta, 2H), 1.53-1.40 (m, 2H), 1.22 (Sirs, 28N), 0.85 (t,3H).

³¹P-NMR: 21.51 (doublet), 19.50 (doublet).

D. 1-On-Hexadecyltrimethyl-3-alendronate (5b)

Compound 4b (300 mg, 0.45 mmol) was dissolved in a mixture of 1,4-dioxane (20 ml) and methanol (5 ml). Then added anhydrous hydrazine and the mixture was stirred at room temperature for 4 hours. The precipitation was collected by vacuum filtration and washed with 1,4-dioxane. The solid is then suspended in ethanol and added a methanolic ammonia solution (3 ml). After stirring for 10 minutes the resulting solid was collected by filtration, washed with ethanol and dried in vacuum. The result was 220 mg HDP-alendronate (5b) in the form of tragamonedas salt. Analysis by FT-IR showed no phthalimides protective group. Mass spectrometry (MS) with elektrorazpredelenie: m/e 532 (MN⁺), 530 (MN^-).

EXAMPLE 2

Synthesis of 1-O-hexadecyloxypropyl-3-pamidronate (5A)

1-O-Hexadecyloxypropyl-3-pamidronate get a similar manner (scheme 1), except that 3-phthalimidopropyl acid is used to produce dimethyl-3-phthalimidopropyl-phosphonate (1A). Compound 1A condense 2, receiving trimethylphosphate 3A. Removing the protection as well as to enter the described stages C and D, receive HDP-pamidronate, as shown.

EXAMPLE 3

Synthesis of 1-O-octadecyl-2-O-methyl-sn-glycero-3-alendronate

Prodrugs with lipophilic groups other than hexadecyloxypropyl, is obtained by replacing hexadecyloxypropyl-1-ol on stage And example 1 at various long chain alcohols. For example, the interaction of 1-O-octadecyl-2-O-methyl-sn-glycerol with diphenylphosphite in pyridine followed by treatment with methanol gives 1-O-octadecyl-2-O-methyl-sn-glycerolphosphate. Condensation of this dialkylphosphate with phosphonate 1b with a subsequent removal of the protection stages C and D gives 1-O-octadecyl-2-O-methyl-sn-glycero-3-alendronate. Scheme 2 illustrates the synthesis of other biphosphonate conjugates that do not have a primary amino group in the side chain. In this case, no protection is needed phthalimide group and removing the protection hydrazinolysis.

EXAMPLE 4

Synthesis HDP-amino-olpadronate

Scheme 3 illustrates the synthesis of 1-amino-biphosphonate conjugates. Using compound 2 of example 1, 3-(dimethylamino)propionitrile and methods described in Orlovskii, V.V.; Vovsi, B.A. J. Gen Chem. USSR (Engl. Transl.) 1976, 46:294-296), get timetravel broadcast biphosphonate 3. In demethylation by bromotrimethylsilane, as described at the stage of example 1, receive HDP-amino-olpadronate.

EXAMPLE 5

Synthesis of 1-O-hexadecyloxypropyl-3-succinyl-alendronate

Scheme 4 illustrates inches biphosphonates conjugate, which lipid group attached to the primary amino group of the parental compounds. Tetramethyl-(4-phthalimido-1-hydrobuilder)biphosphonate (2.0 g, 4.4 mmol) was dissolved in 0.2 M methanolic solution of hydrazine (100 ml) and the solution was stirred at room temperature for 3 days. The mixture was concentrated to half its volume, when began the allocation of solids. This solid was filtered and the filtrate concentrated to dryness. Proton NMR showed that this compound represents tetramethyl-(4-amino-1-hydrobuilder)biphosphonate. It was dried over phosphorous pentoxide at 50°With during the night. To a suspension of 1.2 g of this compound in a mixture of pyridine (25 ml) and N,N-dimethylformamide (25 ml) was added 3-succinyl-1-hexadecyloxypropyl (1,76 g, 4.4 mmol). Added dicyclohexylcarbodiimide (2,52 g, 12,21 mmol) and the mixture was stirred at room temperature for 2 days. The mixture was filtered, the filtrate was absorbed on silica gel and subjected to flash chromatography with an increasing gradient of methanol in dichloromethane (0%-20%), resulting in a received succinylcholine connection. Removed the protection trimethylsilylpropyne in acetonitrile to obtain specified in the title compound, which was purified by crystallization from methanol.

EXAMPLE 6

Synthesis hexadecyloxypropyl and 1-O-sn-glycerol esters of adefovir

To a mixture of adefovir (1,36 g, 5 mmol) and 3-hexadecylamine-1-propanol (1.8 g, 6 mmol) in anhydrous pyridine was added DCC (of 2.06 g, 10 mmol). The mixture was heated under reflux and stirred for 18 hours, then cooled and filtered. The filtrate was concentrated under reduced pressure and the residue was applied to a short column with silica gel. In the elution from the column with a mixture of 9:1 dichloromethane/methanol received hexadecyloxypropyl-adefovir (HDP-ADV) as colorless powder.

To a mixture of adefovir (1,36 g, 5 mmol) and 1-O-octadecyl-sn-glycerol (2,08 g, 6 mmol) in anhydrous pyridine (30 ml) was added DCC (of 2.06 g, 10 mmol). The mixture was heated under reflux and stirred overnight, then cooled and filtered. The filtrate was concentrated under reduced pressure and the residue was applied onto a column of silica gel. In the elution from the column with a mixture of dichloromethane/methanol 9:1 was obtained 1-O-octadecyl-sn-glyceryl-3-adefovir.

EXAMPLE 7

Synthesis hexadecyloxypropyl ether AZT-phosphonate

Phosphonate analogue AZT (3'-azido-3',5'-dideoxythymidine-5'-phosphonic acid) was synthesized using a published method (Hakimelahi, GH; Moosavi-Movahedi, A.A.; Sadeghi, M.M.; Tsay, S-C.; Hwu, J.R. Journal of Medicinal Chemistry, 1995, 38, 4648-4659).

AZT-phosphonate (1.65 g, 5 mmol) suspended in anhydrous pyridine (30 ml), then added a 3-hexadecylamine-1-propanol (1.8 g, 6 mmol) and DC (e 2.06 g, 10 mmol) and the mixture was heated under reflux and stirred for 6 hours, then cooled and filtered. The filtrate was concentrated under reduced pressure and the residue was applied onto a column of silica gel. In the elution from the column with a mixture of dichloromethane/methanol 9:1 received hexadecyloxypropyl ether 3'-azido-3',5'-dideoxythymidine-5'-phosphonic acid.

EXAMPLE 8

Synthesis hexadecyloxypropyl, octadecatetraenoic, octadecylsilane and hexadecafluoro esters cyclo-cidofovir

To a stirred suspension of cidofovir (1.0 g, 3,17 mmol) in N,N-dimethylformamide (N,N-DMF, 25 ml) was added N,N-DICYCLOHEXYL-4-morpholinylcarbonyl (DCMC, 1.0 g, 3.5 mmol). The mixture was stirred overnight to dissolve cidofovir. This clear solution was then placed in an addition funnel and slowly added (30 minutes) to mix hot pyridine solution (25 ml, 60° (C) 1,3-dicyclohexylcarbodiimide (1.64 g,7.9 mmol). The reaction mixture was stirred at 100°C for 16 hours, then cooled to room temperature and the solvent was removed under reduced pressure. The residue was absorbed on silica gel and was purified column flash chromatography using gradient elution (CH₂Cl₂+ Meon). Active in the UV product finally was suirable a mixture of CH₂Cl₂/MeOH/H_{2 O 5:5:1. In the solvent evaporation got 860 mg of a white solid.¹H and³¹P-NMR spectra showed that it is a DCMC-salt of cyclo-cidofovir (yield 44%).}

_{To a solution of cyclo-cidofovir (DCMC-salt) (0.5 g, 0.8 mmol) in anhydrous DMF (35 ml) was added 1-bromo-3-hexadecyloxypropyl (1.45 g, 4 mmol) and the mixture was stirred and heated at 80°C for 6 hours. The solution is then concentrated in vacuo, the residue was absorbed by the silica gel column and purified flash chromatography using gradient elution (CH2Cl2+EtOH). Alkilirovanny product suirable mixture of CH2Cl2/EtOH 90:10. The fractions containing pure product were evaporated to obtain 260 mg HDP-cyclo-cidofovir (yield 55%).}

To a solution of cyclo-cidofovir (DCMC-salt) (1.0 g, 3.7 mmol) in anhydrous DMF (35 ml) was added 1-bromo-3-octadecylsilane (2,82 g, 7.2 mmol) and the mixture was stirred and heated at 85°C for 5 hours. The solution is then concentrated in vacuum and the residue was absorbed by the silica gel column and purified flash chromatography using gradient elution (CH₂Cl₂+Meon). Alkilirovanny product suirable a mixture of CH₂Cl₂/MeOH 9:1. The fractions containing pure product were evaporated to obtain 450 mg ODP-cyclo-cidofovir.

To a solution of cyclo-cidofovir (cCDV) (DCMC-salt) (1.0 g, 3.7 mmol) in anhydrous DMF (3 ml) was added 1-bromo-3-octadecylsilane (3.0 g, 7.9 mmol) and the mixture was stirred and heated at 80°C for 4 hours. The solution is then concentrated in vacuo, the residue was absorbed by the silica gel column and purified flash chromatography using gradient elution (CH₂Cl₂+Meon). Alkilirovanny product suirable a mixture of CH₂Cl₂/MeOH 9:1. The fractions containing pure product were evaporated to obtain 320 mg octadecylsilyl-cCDV.

To a solution of cyclo-cidofovir (DCMC-salt) (0.5 g, 0.8 mmol) in anhydrous DMF (35 ml) was added 1-bromo-3-hexadecan (1.2 g, 4.0 mmol) and the mixture was stirred and heated at 80°C for 6 hours. The solution is then concentrated in vacuum and the residue was absorbed by the silica gel column and purified flash chromatography using gradient elution (CH₂Cl₂+Meon). Alkilirovanny product suirable a mixture of CH₂Cl₂/MeOH 9:1. The fractions containing pure product were evaporated to obtain 160 mg hexadecyl-cCDV.

EXAMPLE 9

Synthesis hexadecyloxypropyl, octadecatetraenoic, octadecylsilane and hexadecafluoro esters of cidofovir

Hexadecyloxypropyl-cyclo CDV from the previous stage was dissolved in 0.5 M NaOH and stirred at room temperature for 1.5 hours. Then added dropwise in 50%aqueous acetic acid to bring the pH to approximately 9. Fallen in sediment HDP-CDV allocated fil is a walkie-talkie, washed with water, dried and then recrystallize (3:1 para-dioxane/water) to obtain the HDP-CDV.

Similarly, octadecatetraenoic, octadecylsilyl and hexadecylamine esters cCDV hydrolyzed using 0.5 M NaOH, and was purified by obtaining the corresponding diesters of cidofovir.

EXAMPLE 10

Synthesis hexadecyloxypropyl ether of cyclo-ganciclovir - phosphonate

Cyclic phosphonate analogue ganciclovir received, using published methods (Reist, E.J.; Sturm, P.A.; Pong, R.Y.; Tanga, M.J. and Sidwell, R.W. Synthesis of acyclonucleoside phosphonates for evaluation as antiviral agents, p. 17-34. In J, C. Martin (ed), Nucleotide Analogues as Antiviral Agents, American Chemical Society, Washingnon, D.C.). After transformation into DCMC-Sol in DMF cyclo-ganciclovir (cGCV)-phosphonate was treated with 1-bromo-3-hexadecyloxypropyl and the mixture was heated at 80°C for 6 hours. In the result of the spin-alkylated product using flash chromatography got HDP-cyclo-GCV-phosphonate.

EXAMPLE 11

Synthesis hexadecyloxypropyl ester of ganciclovir-phosphonate

Obtained in the previous phase of the HDP-cyclo-GCV-phosphonate was dissolved in 0.5 M NaOH and stirred at room temperature in order to turn into acyclic fluids. The solution was neutralized with 50%aqueous acetic acid to precipitate the product, which was recrystallized from a mixture of 3:1 para-dioxane/water.

EXAMPLE 12

1-O-Hexadecyloxypropyl-alendronate inhibited the t caused by dexamethasone apoptosis of osteocytes MLO-Y4

Cells osteocytes MLO-Y4 was pre-treated with 1-O-hexadecyloxypropyl-alendronate (HDP-alendronate) in the indicated concentrations for 1 h, then cells were incubated for 6 hours in the presence and in the absence of dexamethasone (final concentration of 10^-4M). The percentage of dead cells was determined using modernized Trypanosoma blue (Plotkin et al., J Clin Invest 104:1363-1374, 1999). The results are presented in figure 1. Columns represent the average value of ± standard deviation of three independent measurements. Data were analyzed using one-way analysis of variance (ANOVA) (criterion Student-Keuls-Newman (Student-Keuls-Newman)). *p<0,05. HDP-alendronate inhibits induced apoptosis in dexamethasone concentrations from 10^-8up to 10^-5M

EXAMPLE 13

1-O-Hexadecyloxypropyl-alendronate inhibits caused by dexamethasone apoptosis in cells of the cranial vault

Cells of the cranial vault received from newborn mice C57BL/6J and passively in tissue culture. The cells were pre-treated HDP-alendronate at indicated concentrations for 1 h, then cells were incubated for 6 hours in the presence and absence of 10^-4M dexamethasone. The percentage of dead cells was determined using modernized Trypanosoma blue (Plotkin et al., J Clin Invest 104:1363-1374, 1999). The results presented is ENES in figure 2. Columns represent the average value of ± standard deviation of three independent measurements. Data were analyzed by one-way analysis of variance (criterion Student-Keuls-Newman). *p<0,05. Pre-treatment of cells HDP-alendronate at a concentration of 10^-8or more eliminated caused by dexamethasone increase in the percentage of dead cells (p=<0,05). Cells subjected to DEVD (protein inhibitor of apoptosis) in a concentration of 0.05 μm, and then dexamethasone, showed no increase in the percentage of dead cells, demonstrating that DEVD blocks caused by dexamethasone apoptosis.

EXAMPLE 14

Inhibition of bone resorption 1-O-hexadecyloxypropyl-alendronate at ovariectomised rats

Members of groups of female rats of Sprague-Dawley (weighing 250-280 g), which was subjected to bilateral ovarioektomii, treat or disodium salt of 4-amino-1-hydroxybutylidene-1,1-diphosphonic acid or 1-O-hexadecyloxypropyl-3-alendronate, which is administered subcutaneously in doses of transitioning from 0 to 8 mg/kg/day during the period from 4 to 12 weeks. At the time of twelve weeks, rats, including members of the control group, kill and femur of each animal burn. The method of introduction, alternatively, it may be oral. Determine the mass of ash thighs each individual and values srawniwa the t for each group as a measure of bone mass, to determine the relative inhibition of bone loss among treatment protocols. Animals that were treated with 1-O-hexadecyloxypropyl-alendronate, show less loss of bone mass than ovariectomized controls.

EXAMPLE 15

Inhibition of bone resorption 1-O-octadecylsilyl-alendronate in people with osteoporosis

Two groups of women in postmenopausal women treated with placebo or 1-O-octadecylsilyl-alendronate at a dose of from 0.1 to 100 mg/kg/day orally for a period of 2 to 3 years. Members of the treatment groups during the entire course of treatment control bone mineral density, the frequency of vertebral fractures, the progression of vertebral deformities, using radiography, and reduced growth. Do comparisons of measurements at the different groups that are treated, to determine the effectiveness of treatment with alendronate in the group, which is treated. The group that was treated with 1-O-octadecylsilyl-alendronate, has fewer faults, and a lower rate of decline in bone density than the placebo group.

EXAMPLE 16

Stimulation of osteogenesis 1-O-octadecylsilyl-amino-olpadronate people with caused by steroids osteoporosis

Patients with caused by steroids osteoporosis treated with 1-O-octadecylsilyl-amino-what Ladrones or placebo in doses from 0.1 to 100 mg/kg/day orally for a period from 1 month to 1 year. The members of the treated group is continuously controlled during the entire course of treatment bone mineral density, the frequency of vertebral fractures, the progression of vertebral deformities, using radiography, and reduced growth. Do comparisons of measurements at the different groups that are treated, to determine the effectiveness of treatment of 1-O-octadecylsilyl-amino-olpadronate in the group, which is treated. Patients who were treated with 1-O-octadecylsilyl-amino-olpadronate, bone density increased, and fractures compared with placebo treatment.

EXAMPLE 17

Antiviral activity and selectivity phosphonate-nucleotide analogues against human cytomegalovirus (HCMV)

Antiviral analysis of HCMV: antiviral assays for HCMV DNA was performed using DNA hybridization, as described in Dankner, W.M., Scholl, D., Stanat, S.C., Martin, M., Souke, R.L. and Spector, S.A., J. Virol. Methods 21:293-298, 1990. Briefly, subconfluent cells MRC-5 24-hole culture the tablet was pre-treated for 24 hours by different concentrations of drugs in minimum essential medium Needle (E-MEM)containing 2% serum fetal cow (FBS) and antibiotics. The medium was removed and added strains of HCMV in breeding that will result in 3-4 + cytopathic effect (CPE) in the wells without medication after 5 days. Virus AB who was armirovali for 1 hour at 37° With, was aspirated and replaced with solutions of drugs. After 5 days of incubation HCMV DNA was quantitatively determined in three Parallels with the hybridization of nucleic acids using the Kit for determination of antiviral sensitivity against CMV from Diagnostic Hybrids, Inc. (Athens, OH). The medium was removed and was performed by lysis of the cells according to the manufacturer's instructions. After absorption lysate filters Hybriwix^TMhybridized overnight at 60°C. the Filters Hybriwix^TMwashed for 30 minutes at 73°and was made a count in the counter gamma radiation. The results were expressed as EC₅₀(50%inhibitory concentration).

Preliminary experiments were performed with 1-O-hexadecylphosphocholine (HDP) derivative of cidofovir and adefovir, as shown in table 1.

As the results of table 1, 1-O-hexadecyloxypropyl-cyclo-CDV (HDP-cCDV) were more than 900 times more CDV or Ziko-CDV. At higher cytotoxicity index selectivity against HCMV in rapidly dividing cells was more than 59000 compared with selectivity indexes from 1900 to >2100 for non-derivative CDV. Based on these promising preliminary results, conducted further experiments with additional compounds according to the invention. These further experiments described below.

The cytotoxicity of the test compounds in vitro: subconfluent cell lung fibroblasts human (MRC-5, American type culture collection, Rockvill, MD) in a 24-hole tablets were treated with drugs dissolved in environment E-MEM (Gibco BRL, Grand Island, NY)containing 2% serum fetal cow and antibiotics. After 5 days incubation at 37°With the monolayer of cells was visually examined under magnification and counted the concentration of drug that causes 50%reduction in the number of cells.

The data obtained in these experiments are shown in table 2.

Table 2

Inhibition of the replication of cytomegalovirus (CMV) human in the human lung fibroblast MRC-5 in the analysis to reduce DNA

EC₅₀- 50%effective concentration; CC₅₀- 50%cytotoxic concentration; index selectivity CC₅₀/EC₅₀. Results EC₅₀are averages 3-6 definition, except that ADV made one replication in two Parallels.

As the results in table 2, the compounds according to the invention is equally exhibit greater activity and selectivity in comparison with non-derivative with cidofovir, cyclo-cidofovir and adefovir.

EXAMPLE 18

The effect of HDP-cCDV to replicate the se-viruses in vitro

The activity of cidofovir (CDV), cyclo-cidofovir (cCDV) and 1-O-hexadecyloxypropyl-3-cCDV (HDP-cCDV) were tested for antiviral activity in fibroblasts of the foreskin of a man infected with vaccinia virus or vaccinia virus, by measuring the dose-dependent reduction of the cytopathic effect (CPE). Preliminary values EC₅₀for cowpox and ospowiki was determined by analyzing the reduction of CPE in the cells of the foreskin fibroblasts human (HFF). The resulting data are shown in table 3.

As shown in table 3, HDP-cCDV was highly active against vaccinia virus with a value of EC₅₀=0,11 μm compared with 0,97 and 1.8 μm for cCDV and CDV, respectively. In cells infected with spooktinu, HDP-cCDV was extremely effective with a value of EC₅₀less than 0.03 μm, compared to 0.72 and 2.1 for cCDV and CDV, respectively. Based on these promising preliminary data, were investigated steps according to the invention analogs of cidofovir to replicate other orthopox viruses.

Analysis to counter the cytopathic effect (CPE) se-virus: at each concentration of the drugs three wells containing Vero cells, infected orthopox-virus in quantities of 1000 plaque-forming units (PFU) per well, and the other three wells were left uninfected for determining toxicity. The plates were examined and stained after virus infection, untreated cells showed 4+ CPE. To the environment to Avila neutral red and estimated the size of the CPE by the absorption of neutral red at 540 nm. 50%inhibitory (EC₅₀) and 50%cytotoxic (CC₅₀) concentration was determined from the graph of the dose - response. The results are shown in table 4.

EC₅₀- 50%effective concentration; CC₅₀- 50%cytotoxic concentration for Vero cells; index selectivity CC₅₀/EC₅₀; abbreviations are the same as in table 2. The results represent average values of three measurements.

As table 4 shows, the compounds according to the invention is significantly more active than non-derivative CDV or cCDV, against cowpox, ospowiki and different strains of smallpox.

EXAMPLE 19

Step 1-O-hexadecyloxypropyl-3-adefovir (HDP-ADV) on the replication of HIV-1 in vivo

Preliminary experiments on the inhibition of replication of HIV-1 compounds according to the invention is performed as described below. The test drugs were performed as previously described in the Larder et. al., Antimicrobial Agents &Chemotherapy, 34:436-441, 1990. Cells NT-6S infected with HIV-1_LAIwere subjected to action of medications, as indicated, and incubated for 3 days at 37°C. the Cells were fixed crystal violet for visualization painted plaques. Antiviral activity was assessed as the percentage of control plaques (without medication)identified in the samples treated with medication. EC₅₀is a micromolar concentration, which reduces the number of plaques by 50%. The activity of adefovir is compared with the activity of AZT (zidovudine) and 1-O-hexadecyloxypropyl-3-adefovir (HDP-ADV) in cells NT-6S, HIV-1-infected. The results are shown in table 5.

Adefovir showed moderate activity with a value of EC₅₀=16 μm. AZT has been very active, as expected (EC₅₀=0,007 μm), but the HDP-ADV proved to be the most active of these three compounds with the value EC₅₀=0,0001 μm, i.e. more than five log more than adefovir as such. Based on these promising preliminary data, conducted further experiments, as described below.

Antiviral analysis of HIV-1: the effect of antiviral compounds on HIV replication in HeLa cells HT4-6C expressing CD-4 was determined by analyzing the reduction of plaques (Larder, B.A., Chesebro, B. and Richman, D.D. Antimicrob. Agents Chemother., 34:436-441, 1990). Briefly, monolayers of cells HT4-6C infected 100-300 plaque-forming units (PFU) of virus per well in 24-hole tablets for microdesmidae. Added various concentrations of drug to the culture medium, the medium Needle, modified by way of Dulbecco that contained 5% fetal serum cows and antibiotics as described above. After incubation for 3 days at 37°With monolayers were fixed with 10%formaldehyde solution in phosphate buffer solution (PBS) and were stained with 0.25%crystal violet for visualization of viral plaques. Antiviral activity was assessed as the percentage of control plaques identified in the samples treated with medication. Cytotoxicity was evaluated by the method described in Hostetler et al., Antiviral Research, 31:59-67, 1996. The results are shown in table 6.

Table 6

Inhibition of HIV replication in cells NT-6S reduction of plaques

EC₅₀- 50%effective concentration; CC₅₀- 50%cytotoxic concentration; index selectivity CC₅₀/EC₅₀. Value EC₅₀are the average couple, the ex experiments.

How easy it is evident from the results of table 6, the connection according to the invention is 1-O-hexadecyloxypropyl-3-ADV is more active and selective than adefovir.

EXAMPLE 20

Action analogs of cidofovir on the replication of herpes virus

Antiviral analysis in relation to herpes virus-1 (HSV-1): subconfluent cells MRC-5 24-hole culture plates were inoculable by removing the medium and adding the virus HSV-1 at a dilution that will give 3-4 + CPE in the wells without medication for 20-24 hours. Virus was absorbed for 1 hour at 37°With, was aspirated and replaced with various concentrations of drugs in the E-MEM containing 2% fetal serum cows and antibiotics. After about 24 hours of incubation HSV DNA was quantitatively determined in three Parallels with the hybridization of nucleic acids using the Kit for determination of antiviral sensitivity against HSV from Diagnostic Hybrids, Inc. (Athens, OH). The medium was removed and was performed by lysis of the cells according to the manufacturer's instructions. After absorption lysate filters Hybriwix^TMhybridized overnight at 60°C. the Filters Hybriwix^TMwashed for 30 minutes at 73°and was made a count in the counter gamma radiation. Cytotoxicity was evaluated as described in example 17. Thus obtained values EC₅₀and CC₅₀shown in table 7.

Table 7

Inhibition of HSV replication of human rights in the human lung fibroblast MRC-5 (analysis to reduce DNA)

Abbreviations same as in table 2. EC₅₀- 50%effective concentration; CC₅₀- 50%cytotoxic concentration; index selectivity CC₅₀/EC₅₀. Value EC₅₀represent the average values of the two pilot is having except for the value of HDP-CDV, which represents one measurement made in two Parallels.

As shown in table 7, all of the compounds according to the invention are more active against HSV-1 than the nucleotide-phosphonates - derivative, that is, cidofovir or cyclo-cidofovir.

Although the foregoing invention is described in sufficient detail by way of illustration and examples for clarity and understanding, those of ordinary skill in this field will be apparent in light of the teachings of this invention that can be made certain changes and modifications without departure from the essence of the invention and scope of the claims.

1. Phosphonate compound having the structure:

where R₁and R₁' independently represent-H, possibly substituted O(C₁-C₂₄)alkyl, -O(C₁-C₂₄)alkenyl, -O(C₁-C₂₄)acyl, -S(C₁-C₂₄)alkyl, -S(C₁-C₂₄)alkenyl or-S(C₁-C₂₄)acyl, where at least one of R₁and R₁' is not-H, and where specified alkenyl or acyl may have from 1 to about 6 double bonds;

R₂and R₂' independently represent-H, possibly substituted-O(C₁-C₇)alkyl, -O(C₁-C₇)alkenyl, -S(C₁-C₇)alkyl, -S ( 1-C₇)alkenyl, -O(C₁-C₇)acyl, -S(C₁-C₇)acyl, -N(C₁-C₇)acyl, -NH(C₁-C₇)alkyl, -N((C₁-C₇)alkyl)₂, oxo, halogen, -NH₂HE or SH;

R₃is a phosphonate derivative of the nucleoside or biphosphonate;

X represents

L represents a valence bond or a bifunctional linking molecule of the formula-J-(CR₂)_t-G-, where t is an integer from 1 to 24, J and G independently represent-O-, -S-, -C(O)O - or-NH-, a R represents-H, a substituted or unsubstituted alkyl or alkenyl;

m is an integer from 0 to 6; and

n is 0 or 1.

2. Phosphonate compound according to claim 1, where R₃is biphosphonate.

3. Phosphonate compound according to claim 2, where biphosphonate represents alendronate, etidronate, tiludronate, ibandronate, EB-1053, pamidronate, olpadronate, amino-olpadronate, clodronate or risedronate.

4. Phosphonate compound according to claim 1, where R₃is a phosphonate derivative of antiviral nucleoside.

5. Phosphonate compound according to claim 4, where the specified phosphonate derivative represents adefovir, cidofovir, cyclo-cidofovir or tenofovir.

6. Phosphonate compound according to claim 4, where the specified phosphonate derivatives of the same is a derivative of azidothymidine (AZT).

7. Phosphonate compound according to claim 1, where R₃is a phosphonate-derived anticancer nucleoside.

8. Phosphonate compound according to claim 7, where said phosphonate is a derivative of cytosine-arabinoside, gemcitabine, 5-ftordezoksiuridin-riboside, 5-ftordezoksiuridin-deoxyribose, 2-chloromethoxybenzenes, fludarabine or 1-β-D-arabinofuranosyl-guanine.

9. A method of treating osteoporosis in a mammal, wherein the subject in need this, introducing an effective amount biphosphonates connection according to claim 2.

10. A method of increasing bone mineral density, wherein the subject in need this, introducing an effective amount biphosphonates connection according to claim 2.

11. The way to prevent apoptosis of osteoblasts and osteocytes in a mammal, wherein the subject in need this, introducing an effective amount biphosphonates connection according to claim 2.

12. A method of treating a viral infection in a mammal, wherein the subject in need this, introducing an effective amount of phosphonate compound according to claim 4.

13. The method of treatment of a growing neoplasm in a mammal, wherein the subject in need this, introducing an effective amount of phosphonate compound according to claim 7.

14. Method of modulating cell proliferation, wherein the subject in need is semusa, introducing an effective amount of phosphonate compound according to claim 7.

15. The pharmaceutical composition intended for the treatment of osteoporosis, increase bone mineral density or preventing the apoptosis of osteoblasts and osteocytes, containing biphosphonate compound according to claim 2 and a pharmaceutically acceptable carrier for him.

16. The pharmaceutical composition intended for the treatment of viral infections, containing phosphonate compound according to claim 4 and a pharmaceutically acceptable carrier for him.

17. The pharmaceutical composition intended for the treatment of growing neoplasm or modulate cell proliferation, containing phosphonate compound according to claim 7 and a pharmaceutically acceptable carrier for him.