| PATENT ASSIGNEE'S COUNTRY | Japan |
| UPDATE | 05.00 |
| PATENT NUMBER | This data is not available for free |
| PATENT GRANT DATE | 02.05.00 |
| PATENT TITLE |
Amidite derivatives and oligonucleotide derivatives |
| PATENT ABSTRACT | The present invention provides a compound of general formula (I): ##STR1## wherein X represents group (II) or (III): ##STR2## wherein Y represents a leaving group and Z represents an oligonucleotide. The compound can specifically transfer oligonucleotides to cells which specifically recognize a specified saccharide construction. Accordingly, the compound can be used as an antiviral agent or an antitumor agent |
| PATENT INVENTORS | This data is not available for free |
| PATENT ASSIGNEE | This data is not available for free |
| PATENT FILE DATE | 22.12.97 |
| PATENT CT FILE DATE | 29.03.96 |
| PATENT CT NUMBER | This data is not available for free |
| PATENT CT PUB NUMBER | This data is not available for free |
| PATENT CT PUB DATE | 03.10.96 |
| PATENT FOREIGN APPLICATION PRIORITY DATA | This data is not available for free |
| PATENT REFERENCES CITED | This data is not available for free |
| PATENT PARENT CASE TEXT | This data is not available for free |
| PATENT CLAIMS |
We claim: 1. A compound of general formula (I): ##STR17## in which X is group (II): ##STR18## (in which Y is a leaving group) or group (III): ##STR19## (in which Z is an oligonucleotide or a nucleotide derivative in which one or two of the oxygen atoms at a phosphoric ester bonding site are substituted by other atoms or groups as shown by the following formula: ##STR20## wherein A.sup.1 and A.sup.2 are selected from the following combinations: ______________________________________ A.sup.1 A.sup.2 ______________________________________ --OH .dbd.O .dbd.O --CH.sub.3 --OH .dbd.S --SH .dbd.S .dbd.O --O-Alkyl .dbd.S --CH.sub.3 .dbd.O --NH-Alkyl .dbd.O --BH.sub.3 ), ______________________________________ T.sup.1 is --(CH.sub.2)s-- (in which s represents an integer between 2 and 10), or (CH.sub.2 CH.sub.2 O)t--(CH.sub.2).sub.2 -- (in which t represents an integer between 1 and 3), T.sup.2 is --(CH.sub.2)u-- (in which u represents an integer between 2 and 10), --(CH.sub.2 CH.sub.2 O)v--(CH.sub.2).sub.2 --(in which v represents an integer between 1 and 3), or group (IV): ##STR21## (in which T.sup.1 * and T.sup.1 ** are each as defined above for T.sup.1, and n*, p*, q*, T.sup.3 *, T.sup.4 * and F.sup.3 are each as defined below for n, p, q, T.sup.3, T.sup.4 and F.sup.1, where each group and its asterisk-labeled counterpart can be the same or different), T.sup.3, T.sup.4 and T.sup.5, which may be the same or different, each represent --CONH--, --NHCO-- or --O--, provided that when either one of T.sup.3, T.sup.4 and T.sup.5 represents --O--, other two groups represent a group other than --O--, F.sup.1 and F.sup.2, which may be the same or different, each represent a monosaccharide selected from the group consisting of galactose, glucose and galactosamine, or a derivative thereof, or a disaccharide consisting of the monosaccharide and/or the derivative thereof, wherein a hydroxyl group(s) which does not participate in any reactions in the monosaccharide, the derivative thereof and the disaccharide can be protected, and a hydroxyl (group(s) which does not participate in any reactions in the monosaccharide, the derivative thereof, and the disaccharide can be protected, m represents an integer between 0 and 10, n represents an integer between 0 and 4, p represents an integer between 0 and 4, q represents an integer between 0 and 4 and r represents an integer 0 or 1. 2. The compound according to claim 1, wherein s represents an integer between 2 and 8, t represents 2, v represents 2, T.sup.3, T.sup.4 and T.sup.5 represent --CONH--, F.sup.1 and F.sup.2, which may be the same or different, each represent galactose, galactosamine, N-acetylgalactosamine, lactose, lactosamine or N-acetyllactosamine, m represents an integer 0 or between 2 and 10, n represents an integer 0, 1 or 2. P represents an integer 0, 1 or 2, q represents an integer 0, 1 or 2, and r represents an integer 1. 3. A compound of general formula (1a): ##STR22## in which X is group (II): ##STR23## (in which Y is a leaving group) or group (III) ##STR24## (in which z is an oligonucleotide or a nucleotide derivative in which one or two of the oxygen atoms at a phosphoric ester bonding site are substituted by other atoms or groups as shown by the following formula: ##STR25## wherein A.sup.1 and A.sup.2 are selected from the following combinations: ______________________________________ A.sup.1 A.sup.2 ______________________________________ --OH .dbd.O .dbd.O --CH.sub.3 --OH .dbd.S --SH .dbd.S .dbd.O --O-Alkyl .dbd.S --CH.sub.3 .dbd.O --NH-Alkyl .dbd.O --BH.sub.3 ), ______________________________________ T.sup.1 is --(CH.sub.2)s-- (in which s represents an integer between 2 and 8), or --(CH.sub.2 CH.sub.2 O).sub.2 --(CH.sub.2).sub.2 --, T.sup.2 is --(CH.sub.2)u-- (in which u represents an integer between 2 and 8), --(CH.sub.2 CH.sub.2 O).sub.2 --(CH.sub.2)--, or group (IVa): ##STR26## (in which T.sup.1 * and T.sup.1 ** are as defined for T.sup.1, and F.sup.3 is as defined thereinafter for F.sup.1, but can be the same as or different from T.sup.1 and F.sup.3 respectively), F.sup.1 and F.sup.2, which may be the same or different, each represent a monosaccharide selected from the group consisting of galactose and galactosamine, or a derivative thereof, or a disaccharide consisting of the monosaccharide and/or the derivative thereof, wherein a hydroxyl group(s) which does not participate in any reactions in the monosaccharide, the derivative thereof and the disaccharide can be protected, and m is an integer between 3 and 9. 4. The compound according to claim 3, wherein F.sup.1 and F.sup.2, which may be the same or different, each represent galactose, galactosamine, N-acetylgalactosamine, lactose, lactosamine or N-acetyllactosamine. 5. The compound according to any one of claims 1 to 4, wherein X represents group (II). 6. The compound according to claim 5, wherein Y represents an diisopropylamino group or a morpholyl group. 7. The compound according to any one of claims 1 to 4, wherein X represents group (III). 8. The compound according to claim 7, wherein Z is selected from the group consisting of an oligodeoxyribonucleotide and an oligoribonucleotide and their phosphorothioate derivatives and methylphosphate derivatives. 9. The compound according to claim 8, wherein Z is an antisense oligonucleotide. 10. The compound according to claim 8, wherein Z is selected from the sequences consisting of SEQ ID Nos: 1,2 and 3. 11. A pharmaceutical composition comprising the compound according to claim 1. 12. The pharmaceutical composition according to claim 11, which is used for a medicament selected from the group consisting of a therapeutic agent for a malignant tumor, an anti-viral agent, an antirheumatic agent, an anti-inflammatory agent, an anti-allergic agent, an immunosuppressive agent, a circulatory function improving agent and an endocrine function improving agent. 13. A method for treating a disorder selected from the group consisting of a malignant tumor, a viral infection, an inflammatory disorder, an allergic disorder, an immune disorder, a circulatory disorder and an endocrine disorder, comprising administrating to an animal including a human the compound according to claim 1. 14. A method for manufacturing a medicament selected from the group consisting of a therapeutic agent for a malignant tumor, an anti-viral agent, an antirheumatic agent, an anti-inflammatory agent, an anti-allergic agent, an immunosuppressive agent, a circulatory function improving agent and an endocrine function improving agent, which comprises admixing the compound of claim 1 with a carrier therefor. 15. A medicament selected from the group consisting of a therapeutic agent for a malignant tumor, an anti-viral agent, an antirheumatic agent, an anti-inflammatory agent, an anti-allergic agent, an immunosuppressive agent, a circulatory function improving agent and an endocrine function improving agent, which comprises the compound of claim 1 and a carrier therefor. -------------------------------------------------------------------------------- |
| PATENT DESCRIPTION |
BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to amidite derivatives having a monosaccharide or a derivative thereof at their terminals. In particular, the present invention relates to oligonucleotide derivatives in which oligonucleotides are introduced into said amidite derivatives. 2. Description of the Related Art In recent years, attempts have been made to suppress the expression of targeted genes using oligonucleotides, specifically antisense oligonucleotides. However, it was found that when administered directly into the body, the oligonucleotides were readily decomposed in the blood, or the greater portion was readily excreted in the urine. Moreover, the nucleotides were decomposed or excreted without being incorporated into the targeted cells of lesioned organs. To resolve these problems, it has been reported that the formation of a conjugate of asialoorosomucoid with poly-L-lysine yields a complex which ionically interacts with the antisense oligonucleotide of human hepatitis B virus and the ionic complex enhanced the inhibitory effect of the antisence oligodeoxynucleotide on the biosynthesis of viral protein significantly (G. Y. Wu and C. H. Wu (1992) J. Biol. Chem. 267, 12436) and that the chloramphenicol acetyltransferase gene can be transferred into and expressed in the liver using a similar complex (G. Y. Wu and C. H. Wu (1991) Biotherapy 3, 879). Techniques used in these reports are described in WO 93/04701 and 92/20316. Furthermore, it is reported in WO 93/19768 that a complex formed between DNA and a saccharide derivative, which was covalently coupled with a molecule to intercalate into DNA by inserting in the double helix structure (i.e. intercalator) was incorporated into a cell which specifically recognized the saccharide such that it was useful for the efficient expression of genetic information. Nakai et al intravenously injected an antisense nucleic acid complex, and a simulation of the amount delivered into the liver showed that this type of non-covalently bonded complex easily dissociates in the blood to preclude any significant transfer into the liver (D. Nakai, T. Seita and Y. Sugiyama (1995) Pharm. Tech. Japan, 11, 27). On the other hand, it is known that a compound in which galactose is introduced into carboxymethylated dextran (M. Nishikawa et al. (1993) Pharmaceutical Research 10, 1253), and a compound in which galactose is introduced into poly-L-glutamic acid are selectively distributed in hepatocytes (H. Hirabayashi et al (1994) Proceedings of the General Presentation of the 144th Annual Meeting of Japan Pharmacological Association 30(6) 15-4). SUMMARY OF THE INVENTION The present inventors have found that a complex of an oligonucleotide with an amidite derivative having a monosaccharide residue at its terminal is delivered to a specific organ and suppresses expression of a specific gene in cells of the organ. The present invention is based on these findings. Accordingly, an objective of the present invention is to provide compounds which can incorporate oligonucleotides into organ cells, particularly into hepatocytes. Another objective of the present invention is to provide amidite derivatives which are useful for synthesis of the compounds. A compound of the present invention can be represented by general formula (I): ##STR3## in which X is group (II): ##STR4## (in which Y is a leaving group) or group (III): ##STR5## (in which Z is an oligonucleotide or its derivative), T.sup.1 is --(CH.sub.2)s-- (in which s represents an integer between 2 and 10), or (CH.sub.2 CH.sub.2 O)t--(CH.sub.2).sub.2 -- (in which t represents an integer between 1 and 3), T.sup.2 is --(CH.sub.2)u-- (in which u represents an integer between 2 and 10), --(CH.sub.2 CH.sub.2 O)v--(CH.sub.2).sub.2 -- (in which v represents an integer between 1 and 3), or group (IV): ##STR6## in which T.sup.1 * and T.sup.1 ** are each as defined above for T.sup.1, and n*, p*, q*, T.sup.3 *, T.sup.4 * and F.sup.3 are each as defined below for n, p, q, T.sup.3, T.sup.4 and F.sup.1, where each group and its asterisk-labeled counterpart can be the same or different, T.sup.3, T.sup.4 and T.sup.5, which may be the same or different, each represent --CONH--, --NHCO-- or --O--, provided that when either one of T.sup.3, T.sup.4 and T.sup.5 represents --O--, other two groups represent a group other than --O--, F.sup.1 and F.sup.2, which may be the same or different, each represent a monosaccharide selected from the group consisting of galactose, glucose and galactosamine, or a derivative thereof, or a disaccharide consisting of the monosaccharide and/or the derivative thereof, wherein a hydroxyl group(s) which does not participate in any reactions in the monosaccharide, the derivative thereof and the disaccharide can be protected, m represents an integer between 0 and 10, n represents an integer between 0 and 4, p represents an integer between 0 and 4, q represents an integer between 0 and 4 and r represents an integer 0 or 1. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows the inhibitory effect of the compounds of the present invention on expression of c-myc protein in HepG2 cells. Lanes 1, 2, 3 and 4 are with compounds of Example 19 (1), Example 22, Example 21 and Example 19 (2), respectively. In all cases, the compounds were added at a concentration of 1.00 .mu.M. FIG. 2 shows the inhibitory effect of the compounds of the present invention on expression of c-my protein in HepG2 cells. Lanes 1 and 8 are with no compound, Lanes 2-4 are with the compound of Example 19 (2), and Lanes 5-7 are with the compound of Example 19 (1). The compounds were added at a concentration of 0.04 .mu.M for the compounds of Lanes 2 and 5, 0.20 .mu.M for the compounds of Lanes 3 and 6 and 1 .mu.M for the compounds of Lanes 4 and 7. FIG. 3 shows the effect of compounds of the present invention (Examples 20 (1) and (2)) on down regulation of epidermal growth factor receptors in a primary culture of hepatocytes isolated from rats. FIG. 4 shows the effect of compounds of the present invention on growth of HepG2 cells. Black circles, black triangles, white circles and white triangles are with compounds of Example 19 (2), Example 19 (1), Example 18 (2) and Example 18 (1), respectively. DESCRIPTION OF THE PREFERRED EMBODIMENTS Compounds of general formula (I) In general formula (I), F.sup.1, F.sup.2 and F.sup.3 are a monosaccharide selected from the group consisting of galactose, glucose and galactosamine, preferably galactose or galactosamine. The monosaccharide may be a derivative thereof. Examples of such derivative include an N- or O-acyl derivative (e.g., N-acetylgalactosamine), an O-alkyl derivative including carboxyalkyl derivatives (e.g., carboxymethyl derivatives), and an ester derivative with acids such as sulfuric acid, phosphoric acid and carboxylic acid (e.g., sulfate ester derivatives), preferably N-acetylgalactosamine. F.sup.1, F.sup.2 and F.sup.3 may also be a disaccharide consisting of the monosaccharide and/or the monosaccharide derivative. Preferable examples of such disaccharide include those having galactose, galactosamine or N-acetylgalactosamine at the non-reducible end, and are preferably lactose, lactosamine and N-acetyllactosamine. In the present invention, the hydroxyl group of the monosaccharide and the derivative thereof and the disaccharide which do not participate in any reactions can be protected. Examples of such protecting group include an acyl group, preferably a straight chain or branched C.sub.1-6 (preferably C.sub.1-4) alkylcarbonyl group, more preferably an acetyl group. Furthermore, in general formula (I), when group X is group (II), it is preferable that non-reacting hydroxyl groups be protected, and when group X is group (III), it is preferable that non-reacting hydroxyl groups not be protected. The monosaccharide and the derivative thereof and the disaccharide herein mean a saccharide in which one of the hydrogen atom(s) of the hydroxyl group(s) (preferably a hydroxyl group in an anomer position) in the saccharide molecule is removed. In this case, bonds between F.sup.1, F.sup.2 and F.sup.3 and T.sup.1, T.sup.2 and T.sup.1 ** can be either an .alpha.-glycosidic linkage or a .beta.-glycosidic linkage. In T.sup.1 and T.sup.2, s and u are integers between 2 and 10, preferably between 2 and 8, and t and v are integers between 1 and 3, preferably 2. The compounds of the present invention may have a group (IV) described above in T.sup.2 of general formula (I). The group represented by general formula (IV) has substantially the same meaning as that represented by general formula (I) without X--(CH.sub.2)m--(T.sup.5)r-- and --F.sup.2. Accordingly, T.sup.1 * and T.sup.1 ** are as defined in T.sup.1, and may be the same as or different from T.sup.1. Furthermore, n*, p* and q* are integers in the range as defined in n, p and q and may be the same as or different from n, p and q. Furthermore, T.sup.3 *, T.sup.4 * and F.sup.3 are as defined in T.sup.3, T.sup.4 and F.sup.1 and may be the same as or different from T.sup.3, T.sup.4 and F.sup.1. In general formula (I), T.sup.3, T.sup.4 and T.sup.5, which may be the same or different, each independently represent --CONH--, --NHCO-- or --O--, preferably --CONH--. Further, when one of T.sup.3, T.sup.4 and T.sup.5 is --O--, the remaining two are not --O--. In general formula (I): m is an integer between 0 and 10, preferably 0 or between 2 and 10, more preferably between 3 and 9, n is an integer between 0 and 4, preferably 0, 1 or 2, more preferably 0, p is an integer between 0 and 4, preferably 0, 1 or 2, more preferably 0, q is an integer between 0 and 4, preferably 1 or 2, more preferably 2, and r is an integer 0 or 1, more preferably 1. When r is 0, --(T.sup.5)r-- is a bond. Examples of leaving groups represented by Y in group (II) include a diisopropylamino group and a morpholyl group (preferably morpholyl-4-yl group). Examples of oligonucleotides represented by Z in group (III) include an oligodeoxyribonucleotide (DNA) and an oligoribonucleotide (RNA). Furthermore, their sequences and number of bases are not limited and can be appropriately determined according to the use of the compounds. Examples of nucleotide derivatives include those in which one or two of the oxygen atoms at a phosphoric ester bonding site are substituted by other atoms or groups as shown by the following formula: ##STR7## Combinations of A.sup.1 and A.sup.2 and names of the resulting derivatives are as follows: TABLE 1 ______________________________________ Combination of A.sup.1 and A.sup.2 and names of derivatives A.sup.1 A.sup.2 Name of derivative ______________________________________ --OH .dbd.O Phosphodiester (natural type) .dbd.O --CH.sub.3 Methyl phosphonate --OH .dbd.S Phosphorothioate --SH .dbd.S Phosphorodithioate .dbd.O --O--R Alkylphosphotriester .dbd.S --CH.sub.3 Methylphosphonothioate .dbd.O --NH--R Alkylphosphoramidite .dbd.O --BH.sub.3 Boranophosnate ______________________________________ In the table, R represents an alkyl group. Further, substitution may occurred at all or a part of the phosphoric ester bonds in the nucleotides and substitution may occurred at an atom(s) or group(s) in each phosphoric ester bond. Examples of oligonucleotide derivatives which can be easily synthesized include natural phosphodiesters and phosphorothioate derivatives. Oligonucleotides represented by Z can be antisense oligonucleotides. Examples of antisense oligonucleotides include those which have antiviral activity, in particular, an antisense oligonucleotide against the surface antigen of hepatitis B virus (HBsAg) (Goodarzi, G. at al (1990) J. Gen. Virol. 71, 3021) and an antisense oligonucleotide against the envelope protein of hepatitis B virus (HBeAg) (Blum, H. E. et al (1991) Lancet 337, 1230). Other examples include (2'-5')oligoadenylate which is known to be responsible for the antiviral activity of interferon and those which suppress expression of cancer genes. Examples of oligonucleotides represented by Z include DNA sequences shown in SEQ ID Nos: 1 to 3. The sequence of SEQ ID No: 1 is a 15 mer oligodeoxynucleotide (sense sequence) having a sequence identical to the base sequence of 5 codons starting from the translation start codon toward the 3' end of messenger RNA derived from the human c-myc gene. The sequence of SEQ ID No: 2 is a 15 mer oligodeoxynucleotide (antisense sequence) having a sequence complementary to the base sequence of 5 codons starting from the translation start codon toward the 3' end of messenger RNA derived from the human c-myc gene. The sequence of SEQ ID No: 3 is a 18 mer oligodeoxynucleotide (antisense sequence) having a sequence complimentary to the sequence between the 33th and the 50th from the 3' end of messenger RNA derived from the rat epidermal growth factor receptor protein. A group of preferable compounds according to the present invention include compounds of formula (I), in which T.sup.1 is --(CH.sub.2)s-- (in which s represents an integer between 2 and 8) or --(CH.sub.2 CH.sub.2 O).sub.2 --(CH.sub.2 ).sub.2 --, T.sup.2 is --(CH.sub.2)u-- (in which u represents an integer between 2 and 8), --(CH.sub.2 CH.sub.2 O).sub.2 --(CH.sub.2 ).sub.2 --, or group (IV) (in which T.sup.1 * and T.sup.1 ** are as defined for T.sup.1 but each can be the same as or different from T.sup.1, and n*, p*, q*, T.sup.3 *, T.sup.4 * and F.sup.3 are as defined thereinafter for n, p, q, T.sup.3, T.sup.4 and F.sup.1, but can be the same as or different from n, p, q, T.sup.3, T.sup.4 and F.sup.1, respectively), T.sup.3, T.sup.4 and T.sup.5 are --CONH--, F.sup.1 and F.sup.2, which may be the same or different, each represent galactose, galactosamine, N-acetylgalactosamine, lactose, lactosamine or N-acetyllactosamine, m is an integer 0 or between 2 and 10, n is an integer 0, 1 or 2. P is an integer 0, 1 or 2, q is an integer 0, 1 or 2, and r is an integer 1. More preferable compounds according to the present invention are compounds expressed by general formula (Ia): ##STR8## in which X is group (II): ##STR9## (in which Y is a leaving group) or group (III) ##STR10## (in which z is an oligonucleotide or a derivative thereof) T.sup.1 is --(CH.sub.2)s-- (in which s represents an integer between 2 and 8) or --(CH.sub.2 CH.sub.2 O).sub.2 --(CH.sub.2).sub.2 --, T.sup.2 is --(CH.sub.2)u-- (in which u represents an integer between 2 and 8), --(CH.sub.2 CH.sub.2 O).sub.2 --(CH.sub.2).sub.2 --, or group (IVa): ##STR11## in which T.sup.1 * and T.sup.1 ** are as defined for T.sup.1, and F.sup.3 is as defined thereinafter for F.sup.1, but can be the same as or different from T.sup.1 and F.sup.1 respectively, and F.sup.1 and F.sup.2, which may be the same or different, each represent a monosaccharide selected from the group consisting of galactose and galactosamine, or a derivative thereof, or a disaccharide consisting of the monosaccharide and/or the derivative thereof, wherein a hydroxyl group(s) which does not participate in any reactions in the monosaccharide, the derivative thereof and the disaccharide can be protected, and m is an integer between 3 and 9. The compounds of the present invention have a monosaccharide or a derivative thereof at their terminals. Accordingly, the compounds of the present invention can deliver a specified sugar structure to cells which specifically recognize it. Preparation of Compounds of General Formula (I) The compounds of general formula (I) in which group X is not group (II) or group (III) but a hydroxyl group can be obtained by one of the following method (1), (2) or (3): (1) A compound of formula (V): ##STR12## (in which R.sup.1 is a halogen atom, a protected or unprotected hydroxyl group, amino group or carboxyl group, T.sup.1-4, F.sup.1, F.sup.2, n, p and q are as defined above, but functional groups not involved in any reactions of F.sup.1 and F.sup.2 are preferably protected) may be reacted with the compound of formula (VI): R.sup.2 --(CH.sub.2)m--R.sup.1 (VI) (in which R.sup.1 is as defined above, R.sup.2 is a protected or unprotected hydroxyl group, and m is as defined above) as follows: to form an amide bond, by a condensation method in the presence of a condensation agent (e.g., dicyclohexylcarbodiimide), by reaction with a mixed acid anhydride in the presence of isobutyl chlorocarbonate or the like, or by reaction with an active ester using hydroxysuccinimide or the like; alternatively to form an ether bond, by a condensation reaction between a corresponding halogen compound and alkoxide. In either case, the usual reaction temperatures and reaction times for the respective method can be applied. The compound of formula (V) above can be obtained by reacting a compound of formula: ##STR13## (in which R.sup.1, n, p and q are as defined above) with a compound of formula (VIII): R.sup.1 --T.sup.1 --F.sup.1 (VIII) (in which R.sup.1, T.sup.1 and F.sup.1 are as defined above) by a condensation reaction or the like as described above, followed by deprotection if necessary. Further, a compound of formula (V) in which T.sup.2 is represented by group (IV) can be obtained by reacting (e.g., condensation) two of the same or two different compounds of formula (VII), occasionally followed by deprotection if necessary, then by reacting the intermediate with a compound of formula (VIII). (2) A compound of formula (IX): ##STR14## (in which R.sup.1, R.sup.2, T.sup.5, m, n, p and q are as defined above) and a compound of formula (VIII) are reacted by a condensation reaction or the like as described above, occasionally followed by deprotection if necessary, to obtain the target compound. The compound of formula (IX) can be obtained by reacting a compound of formula (VII) with a compound of formula (VI) by a condensation reaction or the like as described above, followed by deprotection if necessary. Further, a compound of formula (I) in which T.sup.2 is represented by group (IV) can be obtained by reacting a compound of formula (VII) with a compound of formula (IX) by a condensation reaction or the like as described above, followed by deprotection if necessary, and then reacting the intermediate with a compound of formula (VIII). (3) A compound of formula (X): ##STR15## (in which R.sup.1-5, R.sup.2, m, n, p, q and r are as defined above) and a compound of formula (XI): D--F* (XI) (in which D is a halogen atom, an acyloxy group (e.g., acetoxy group) or CCl.sub.3 C(.dbd.NH)O--, and F* is F.sup.1 or F.sup.2) are glycosylated at a reaction temperature between -20.degree. C. and room temperature for 10 minutes to 24 hours, followed by deprotection if necessary, to obtain the target compound. The compound of formula (X) above can be obtained by reacting a compound of formula (IX) with a compound of formula (XII): R.sup.3 --T.sup.1 --R.sup.2 (XII) (in which R.sup.2, R.sup.3 and T.sup.1 are as defined above) by a condensation reaction or the like as described above, followed by deprotection if necessary. A compound of general formula (I) in which group X is group (II) can be obtained by reacting a compound of general formula (I) in which group X is a hydroxyl group with a compound of formula (XIII): ##STR16## (in which Y is a leaving group) in the presence of an activating reagent (e.g., tetrazole) at a reaction temperature between -20.degree. C. and room temperature for 10 minutes to several hours. A compound of general formula (I) in which group X is group (III) can be obtained by reacting a compound of general formula (I) in which group X is group (II) with a nucleotide using an ordinary DNA synthesis method such as the .beta.-cyanoethylphosphoramidite method. In the .beta.-cyanoethylphosphoramidite method, nucleotides are first immobilized on a solid phase, then coupled with an amidite monomer (in which hydroxyl groups not involved in bonding are preferably protected) activated by an activating agent such as tetrazole, oxidized with an oxidizing agent (e.g., an aqueous iodine solution), and cleaved from the solid phase, and deprotection if necessary. Natural phosphodiester-type oligonucleotides to be immobilized on a solid phase can be obtained in advance by repeating this reaction. Furthermore, phosphorothioate-type oligonucleotides can be synthesized using a reagent which can generate free sulphur atoms in an oxidation reaction (e.g., Beaucage reagent). Furthermore, various phosphoric ester bonds can be formed using amidites in which oxygen atoms at phosphoric acid sites are substituted by various functional groups. For example, a phosphorodithioate-type oligonucleotide can be obtained by oxidizing with sulphur atoms using 5'-dimethoxytrityldeoxynucleoside 3'-(dimethylamino) phosphorothioamidite (W. K. D. Bill et al (1989) J. Am. Chem. Soc. 111, 2321). Furthermore, a methylphosphonate-type phosphoric ester bond can be formed using 5'-methoxytrityldeoxynucleoside 3'-methylphosphonate and mesitylenesulfonyl-3-nitrotriazole (P. S. Miller et al (1983) Nucleic Acid Res. 11, 6225). Furthermore, an ethylphosphotriester-type phosphoric ester bond can be formed using 5'-dimethoxytrityldeoxynucleoside 3'-O-ethyl-N,N-diisopropylphosphoramidite (K. A. Gallo et al (1986) Nucleic Acid Res. 14, 7405). The compounds so synthesized are purified by partition chromatography (e.g., octadecyl silica gel column chromatography), ion-exchange chromatography (e.g., anion-exchange column chromatography), affinity chromatography (e.g., RCA lectin affinity chromatography) or the like. Use of Compounds/Pharmaceutical Compositions The compounds of the present invention have a monosaccharide or a derivative thereof at their terminals. Therefore, the compounds of the present invention can be delivered specifically to cells which recognize a specified sugar structure. Furthermore, the compounds of the present invention can have an oligonucleotide or a derivative thereof at their terminals. This oligonucleotide can be one which can suppress expression of a specified gene in cells of a targeted organ, for example, an antisense oligonucleotide. Accordingly, the compounds of the present invention can be used as therapeutic agents for various diseases. The compounds of the present invention can deliver an antisense oligonucleotide which is effective as an anti-viral agent to hepatic cells infected with viruses to enhance anti-viral activity. Furthermore, the compounds of the present invention can deliver an antisense oligonucleotide which is effective as an anti-malignant tumor agent to cancerous hepatic cells to enhance anticancer activity. Another aspect of the present invention is to provide pharmaceutical compositions comprising the compound of the present invention together with pharmaceutically acceptable carriers. Thus, the pharmaceutical compositions can be used as a therapeutic agent for malignant tumors (e.g., a therapeutic agent for cancers), an anti-viral agent, an antirheumatic agent (e.g., an agent to suppress production of tumor necrosis factor), an anti-inflammatory agent, an anti-allergy agent or an immunosuppressive agent (e.g., an agent to inhibit migration of immunocompetent cells to inflammatory sites), an agent to improve circular functions (e.g., agents to inhibit growth of vascular smooth cells associated with re-obstruction of coronary vessels), an agent to improve endocrine functions (e.g., agents to inhibit abnormal hormone secretion), or a therapeutic agent for diseases which are caused by abnormal expression or functional abnormality of specific proteins and of which symptoms can be improved by suppressing expression of the proteins (e.g., an agent to suppress abnormal expression of receptor proteins of cells). If the pharmaceutical composition is a therapeutic agent for malignant tumors, Z in formula (I) can be an antisense oligonucleotide to suppress expression of cancer genes. If the pharmaceutical composition is an anti-viral agent, Z in formula (I) can be an antisense oligonucleotide having antiviral activity. A pharmaceutical composition of the present invention can be administered to human and other animals either orally or non-orally (e.g., intravenous and intramuscular injection, and subcutaneous, rectal, endermic and nasal administration). The compounds of the present invention can be prepared into suitable dosage form depending on their use, such as tablets, capsules, granules, powders, pills, grains and troches for oral administration, injectable solutions for intravenous or intramuscular injections, formulations for rectal administration, oily suppositories and water-soluble suppositories. These various pharmaceutical preparations can be prepared by ordinary methods using customary excipients, bulking agents, binders, wetting agents, disintegrating agents, surfactants, lubricating agents, dispersing agents, buffering agents, preservatives, solubilizing agents, antiseptics, flavor/odor controlling agents, analgesic agents, stabilizers or the like. Examples of the nontoxic additives to be used include lactose, fructose, glucose, starch, gelatin, magnesium carbonate, synthetic magnesium silicate, talc, magnesium stearate, methylcellulose, carboxymethyl cellulose or salts thereof, gum arabic, polyethylene glycol, syrup, vaseline, glycerin, ethanol, propylene glycol, citric acid, sodium chloride, sodium sulfite and sodium phosphate. If necessary, effective components other than the compounds of the present invention can be added. The particular dose for each individual patient is determined as a function of usage, age and sex of the patient and severity of symptoms; however, a daily dose for an adult is generally between about 0.05 and 250 mg, preferably between about 0.5 and 50 mg, which can be administered as a single dose or divided into several doses. In this specification, the term "therapy" means both the treatment and the prevention of diseases. Another aspect of the present invention is to provide a method for treating a disease selected from the group consisting of a malignant tumor, a viral infection, an inflammatory disease, an allergic disease, an immune disease, a circulatory disease and an endocrine disease comprising administrating the compound of the present invention to an animal (e.g., a mammal) including a human. Another aspect of the present invention is to provide use of the compound of the present invention for manufacturing a medicament selected from the group consisting of a therapeutic agent for malignant tumors, an anti-viral agent, an antirheumatic agent, an anti-inflammatory agent, an anti-allergic agent, an immunosuppressive agent, an agent to improve circulatory functions and an agent to improve endocrine functions, and use of the compound of the present invention for a medicament selected from the group consisting of a therapeutic agent for malignant tumors, an anti-viral agent, an antirheumatic agent, an anti-inflammatory agent, an anti-allergic agent, an immunosuppressive agent, an agent to improve circulatory functions and an agent to improve endocrine functions. |
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