TW200538138A - Novel sugar chain-containing carbosilane dendrimer and method for producing the same, dengue virus infecton inhibitor, antiviral agent, and target substance for screening anti-HIV drug - Google Patents

Abstract

A sugar chain-containing carbosilane dendrimer shown by the following general formula, the following general formula, or the following general formula, a method for producing the same, a dengue virus infection inhibitor, an antiviral agent, and a target substance for screening an anti-HIV drug. According to the present invention, a novel sugar chain-containing carbosilane dendrimer and a method for producing the same, a dengue virus infection inhibitor, an antiviral agent, and a target substance for screening an anti-HIV drug containing the sugar chain-containing carbosilane dendrimer as an active ingredient can be provided.

Description

200538138 IX. Description of the invention: [Technical field to which the invention belongs] The present invention relates to a novel sugar-carrying carbon containing a substance that can be used as a target substance for the screening of dengue virus infection inhibitors, antivirals or anti-rhenium IV agents. Silane dendrimer and manufacturing method thereof, and target for screening dengue virus infection inhibitor, antiviral agent and anti-thorium IV agent using the above-mentioned sugar chain-carrying carbon stone yam dendrimer as an active ingredient Material person. [Prior art]

Dengue fever is an acute infectious disease of dengue virus. From its clinical characteristics, it can be classified as classical dengue fever (hereinafter also referred to as CDF) with good prognosis. Hemorrhagic fever (dengue hemorrhagic syndrome (hereinafter also referred to as DHF)) and the most severe dengue shock syndrome (hereinafter also referred to as DSS) characterized by shock. C D F is a symptom of fever, headache, low back pain, flushing of the face, conjunctival congestion, etc. that occur suddenly at about 40 ° C after a latency period of 3 to 9 days. It is accompanied by intense joint pain and muscle pain in the whole body. In addition, symptoms of digestive organs or upper respiratory tract inflammation later appeared. However, these symptoms are defined by the <patient> themselves, and will naturally heal / recover after time has passed. On the other hand, D H F and D S S also had symptoms almost the same as CD D F, but the difference was that obvious bleeding tendency or shock-like symptoms appeared at 2-6, severe feeling of collapse or weakness, and the state quickly deteriorated. Dengue fever is widely distributed in tropical places around the world and is highly contagious. It is known that about 80% of the population was infected during the epidemic. Patients on Earth have reached 312XP / Invention Manual (Supplement) / 94-08 / 9411233 8

200538138 2 million people per year (W Η 0), the epidemic area and the number of patients have been expanding year by year. Furthermore, in recent years, DSS has not occurred in most places in the past. The fact that the death rate of hemorrhagic fever is as high as 40% or more can also be a symptom. The symptoms have been identified as re-infection. The countermeasures are extremely important in public health Problem. However, there is very little information about the target tissues of infection, the names of the early stages of infection, and the molecules and genes involved in toxic interactions. Actually, drugs effective for dengue and dengue hemorrhagic fever have been found. Regarding dengue, although research and development of attenuated vaccines, inactive vaccines, subunit vaccines, vaccines, DNA vaccines, etc. are in progress, due to the problem of effective response, they have not yet reached practicality. Therefore, there is a need for a novel inhibitor of dengue virus infection. Furthermore, viruses such as influenza virus have various substances on the surface. These proteins recognize sugar chains and the like in an organism, and then infect the organism with the sugar chains. As an antiviral agent against viruses such as influenza virus, the re-published patent (International Publication No. W 0 2/0 0 2 5 8 8) reveals the carbon containing sugar chains. Antiviral agents against viruses such as epidemic viruses are still a minority and are still true. Therefore, there is a need for a novel antiviral agent against influenza viruses and the like. On the other hand, mannose is a component that controls various life glycoproteins in the body and is highly accumulated. In particular, the surface of GP 1 2 0 molecular system is a glycoprotein with an N-linked sugar bond 312XP / Invention Specification (Supplements) / 94-08 / 941] 2338 A large amount of DHF and known The disease has not yet been joined by fever vaccine recombinant epidemics or paraproteins, although there is a cold virus activity on the lutein IV, especially 6.200538138 which has most of the high mannose type sugar chains. Therefore, if the accumulation of mannose can be performed, it is possible to mimic the g p 120 molecule and become a target substance for screening anti-muscarinic IV agents. (Patent Document 1) Republished Patent (International Publication No. W 0 0 2/0 0 2 5 8 8)

Accordingly, an object of the present invention is to provide a novel carbosilane-containing dendrimer having a sugar chain, which can be used as a target substance for the screening of dengue virus infection inhibitors, antiviral agents, or anti-thorium IV agents, and its manufacture. A method, and a substance for screening dengue virus infection inhibitors, antiviral agents, and anti-thorium IV agents using the above-mentioned carbonaceous yakiden dendrimers with sugar bonds as active ingredients. [Summary of the Invention] Under the above-mentioned facts, the inventors of the present case and other intensive studies have found that a carbosilane dendrimer formed by carrying a sugar chain with a specific structure can be used as an inhibitor of dengue virus infection, an antiviral agent, or an anti-thorium The target substance for sieving of the IV agent has completed the present invention. That is, the present invention (1) provides the following general formula (1) (R 1), “S i {-R 2-S i (R 6) 1 [R 3-S i (R7) I, (R 4 -S-R 5-A) 3-, &lt;] 3-.} N (1) (where R |, R6 and R7 are alkyl, phenyl, vinyl and olefin having 1 to 6 carbon atoms Any one of propyl groups may be the same or different, and R2, R3, R4, and R5 are any of alkylene, phenylene, and alkenyl groups having 1 to 6 carbon atoms, and may be the same Or different, A is the following chemical formula

312XP / Invention Specification (Supplement) / 94-08 / 94] 1233 S 7.200538138 The erythrocyte glycoside ester (parag 1 〇boside) derivation group, or alpha mannosyl group, or ~ 8 mannose chains formed by the mannose of the oligosaccharides at the terminal 1 position of the hydroxyl group to remove the hydrogen atom, η is an integer of 0 to 3, η is an integer of 1 to 4, m + n two 4, In other words, k and 1 are any of 0 to 2, k and 1 may be the same or different), and the carbon silicon sintered dendrimers with sugar chains shown in FIG. Furthermore, the present invention (2) provides the following general formula (2) (R1) mSi [-R2-Si (R6) i (R4-S-R5-A) 3-i] n (2)

(In the formula, the definitions of 1 ^, 1 ^, 1 ^, 1 ^, 1 ^, eight, 111, 11, and 1 are as described above.) The carbon fragments carrying sugar chains are shown as follows :): Complete dendrimer . Furthermore, the present invention (3) provides the following general formula (3) (R ^ .Si (R4-S-R5-A) n (3) (wherein R], R4, R5, A, m, and The definition of η is as shown in the above-mentioned carbonaceous yakaya dendrimer carrying sugar chains. Furthermore, the present invention (4) provides any one of the aforementioned inventions (1) to (3). The carbosilane-containing dendrimer having a sugar chain is characterized in that A is the aforementioned erythrocyte glycoside ester derivative group. Furthermore, the present invention (5) provides any one of the aforementioned inventions (_1) to (3). The carbosilane dendrimer carrying a sugar chain according to one item, wherein A in the formula is the above-mentioned α-mannosyl glycosyl group, or from only one containing 2 to 8 mannose chains. The hydroxy group at the terminal 1 position of the oligosaccharide formed by mannose removes the hydrogen atom group.

Furthermore, the present invention (6) provides the sugar chain-carbosilane-containing dendrimer according to any one of the aforementioned inventions (1) to (3), wherein A 8 312XP / invention Instruction (Supplement) / 94-08 / 94112338 200538138 is the aforementioned a mannan glycosyl 'or the following chemical formula (2)

The base shown. Furthermore, the present invention (7) provides a method for producing a carbosilane-containing dendrimer having a sugar chain according to the aforementioned invention (1), which is characterized in that the general formula (4) (R ,) „1Si {-R2-Si (R6) i [R3-Si (R7) R (R4-X) 3-k] 3-i} n (4) (where 1? 1, 1? 2, The definitions of 1? 3, 1? 4, 1? 6, 1? 7, 111, 11, 1 ^, and 1 are as described above, and X represents a halogen atom) The halogen compound represented by the following general formula (5) A-R5 -SY (5)

(In the formula, R5 is as defined above, and A is a sugar chain as described above, or a part or all of the hydroxyl groups of the hydroxyl groups of the sugar chain are substituted with ethenyl, benzyl, and benzyl The sugar chain of any one of the groups, Y represents a reactive detachable protecting group), and the sulfur compound is reacted. Furthermore, the present invention (8) provides a method for producing a carbosilane-containing dendrimer having a sugar chain according to the above-mentioned invention (2), characterized in that the following general formula (6) (R1) ,, ^! {-R2-Si (R6) i (R4-X) 3-i} n (6) (wherein 1 ^, 1 ^ 2, 1 ^, 1 ^ 6, 111, 11 and 1) The definition is as described above, and the halogen compound represented by） represents a halogen atom) reacts with the sulfur compound represented by the aforementioned general formula (5). 9 312XP / Invention Specification (Supplement) / 94-08 / 9411233 8 200538138 Furthermore, the present invention (9) provides a method for producing a carbon silane dendrimer with a sugar chain as described in the aforementioned invention (3) The method is characterized in that the following general formula (7) (R1) mSi (R4-X) n (7) (wherein the definitions of R1, R4, m, and η are as described above, and X represents a halogen atom): The halogen compound is reacted with a sulfur compound represented by the aforementioned general formula (5). In addition, the present invention (10) provides a dengue virus infection inhibitor, characterized in that it contains a carbosilane tree dendrimer having a sugar chain as described in the aforementioned invention (4) as an active ingredient. Furthermore, the present invention (Π) provides an antiviral agent, characterized in that it contains a carbosilane dendrimer having a sugar chain as described in the above invention (4) as an active ingredient. Furthermore, the present invention (1 2) provides a sieving target substance for anti-rhenium IV agent, which is characterized in that it contains a carbon-silicon dendrimer with sugar chains as described in the above-mentioned invention (5) as Active ingredients.

According to the present invention, a novel carbosilane-containing dendrimer having a sugar chain and a method for producing the same, which can be used as a sieving target substance for dengue virus infection inhibitors, antiviral agents, and anti-thorium IV agents, and a manufacturing method thereof, and The above-mentioned sieving dendritic virus infection inhibitors, antiviral agents, and anti-thorium IV agents, which are sugar carbohydrate-carrying dendrimers containing the sugar chains, are effective targets for screening. [Embodiment] The carbosilane-containing dendrimer having a sugar chain shown in the aforementioned general formula (1) of the present invention may be, for example, the following chemical formula D umbbe 1 1 (2) 18 type 10 312XP / invention Instruction (Supplement) / 94-08 / 9411233 8 200538138 Contains carbosilane dendrimers with sugar chains. In the general formula (1) below, R1 is a methyl group, m = 2, n = 2, k = 0, and 1 = 0. In the following chemical formula, a terminal portion shown by three circles represents a sugar chain.

Furthermore, the sugar chain-carrying carbon silane dendrimer shown in the aforementioned general formula (2) of the present invention may be, for example, the following chemical formula D umbbe 1 1 (1) 6 type-carrying sugar chain-carrying carbon Silane dendrimer. In the following chemical formula, a terminal portion shown by an oval is a sugar chain.

In addition, the carbosilane-containing dendrimer having a sugar chain shown in the aforementioned general formula (3) of the present invention may be, for example, B a 1 1 (0) 4 type and F a η (0 ) Type 3 carbosilane dendrimers with sugar chains. In the following chemical formula, the terminal portion shown by the oval is a sugar chain. 11 312XP / Invention Specification (Supplement) / 94-08 / 94] 12338 200538138 s

Fan (0) 3-Sugar In the carbosilane-containing dendrimers with sugar chains of the present invention, Dumbbe 1 1 (2) 1 8 and Du Ni bbe 1 1 (1) 6 Carbosilane dendrimers of sugar chains are preferred because of their good activity as inhibitors of dengue virus infection. 12 3] 2XP / Invention Specification (Supplement) m-〇8 / 94]] 2338 200538138 The carbosilane-containing dendrimer carrying sugar chains of the present invention can be produced by the following method. <Manufacture of halide> The production of the halide represented by the aforementioned general formulae (4), (6), and (7) can be performed by the method of Terunuma et al. (Bull. Chei Soc. Jp., 72 (1999), p2129-2134). <Manufacture of sugar chains>

The sugar chain used in the preparation of a carbosilane dendrimer containing a sugar chain containing a erythroglycoside-derived group is, for example, the implementation of a sugar chain receptor derived from lactose and a sugar chain derived from lactose The glycosylation of the donor can be made after the conversion of the protective group and then thioethylation. Carbosilane containing a sugar chain containing an alpha mannosyl group, or a hydrogen chain group removed from a hydroxyl group at the terminal 1-position of an oligosaccharide formed from mannose containing 2 to 8 mannose chains The sulfur compounds used in the preparation of dendrimers can be based on Zhang et al. (T etrahedr ο η:

Asymmetry, 13, (2002), p243-252). "Introduction of Sugar Bonds and Preparation of Carbon-Stone Burning Dendritic Molecules Carrying Sugar Bonds" After reacting the aforementioned sulfur compounds with functional compounds, carbon-silane branches carrying sugar chains can be prepared by deprotection.状 聚合物。 Shaped polymer. The carboniferous dendrimers containing sugar bonds of the present invention are effective as a dengue virus infection inhibitor system when the sugar bonds are derived from erythrocyte glycoside esters. It also has a carbosilane dendrimer skeleton, which can be used as an antiviral agent for influenza virus temples. In addition, in the case where the sugar bond is composed of mannose, it can be a target substance for screening anti-muscarinic IV agents. 13 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 Among the antiviral agents such as the dengue infection inhibitor and influenza virus invented in this case, the administration method can be selected according to the patient's symptoms and status, such as Oral administration in the form of tablets, granules / powders, syrups, etc., non-oral administration in the form of injections, rectal administration in the form of suppositories, etc.

When the antiviral agents such as the dengue virus infection inhibitor and the influenza virus in this case are administered orally, tablets, tablets, capsules, powders, granules, suspensions, emulsions and syrups can be formed. form. Furthermore, it may take the form of slow release or delayed release of coated particles, multilayer tablets, or fine particles. In these forms, antiviral agents such as dengue virus infection inhibitors and influenza viruses may contain pharmaceutically acceptable binders, sweeteners, disintegrants, diluents, artificial flavors, coatings, preservation Agents, lubricants, and / or effect retarders. For parenteral administration, rectal administration in the form of suppositories or the like is also acceptable. A suitable suppository can also be prepared by mixing the active substance with a solid at room temperature and melting a non-irritating excipient in the intestine. Inhibitors of dengue virus infection and antiviral agents such as influenza virus in this case may also be inhaled sprays or those having a transdermal administration form such as ointment. For example, the inhalation spray may be formed into a solution, suspension, or emulsion, and a low-toxicity inhalable spray containing carbon dioxide, nitrous oxide, or the like may be used. On the other hand, the preferred form for transdermal administration is, for example, the form of an emulsion, ointment, gel, jelly, elixir, suspension, or emulsion. These may also contain pharmaceutically acceptable binders, diluents, disintegrating agents, preservatives, lubricants, dispersing agents, suspending agents, and / or emulsifying agents. It is also possible to manufacture the antiviral agent 14 312XP / Invention Manual (Supplement) / 94-08 / 94112338 200538138 antiviral agent of the dengue virus infection inhibitor and the influenza virus of the present case by various generally known methods. For example, the carbosilane dendrimers with sugar chains on the active ingredients are crushed, crushed, combined, dispersed, dissolved, suspended, mixed, and combined with carriers, adjuvants, diluents, or excipients. , Emulsified, or homogenized to prepare. Furthermore, it is also possible to manufacture by combining one or more of these steps. Regarding the inhibitors of dengue virus infection and the antiviral agents such as influenza virus in this case, the content of the active ingredients is not particularly limited. For example, the carbon-containing sugar-bonded carbon-containing dendrimer molecule of the active ingredient can be formulated to a concentration of 500 to 100 mg / person / day. Of course, the dosage of a patient is determined by considering the patient's age, sex, weight, etc., and the diagnosis is made by the attending physician according to the patient's symptoms and status. Preferably, it is administered within a range of 10 to 100 mg / kg according to the weight of the patient. Next, the present invention will be described in more detail by way of examples, but this is merely an example and does not limit the present invention. (Manufacturing example) 〈Synthesis of carbosilane dendritic polymer skeleton: D umbbe 1 1 (1) 6-B r &gt; According to the following reaction formula, a carbon silane dendrimer with Dumbbe 1 1 (1) 6 type was prepared. Molecular skeleton II compound.

Dumbbel l (l) 6-Br

Me

MgBr

Pt cat. ΒΓ

Si Me (i) CHSi-CI Me

(ii) Guangya BH2 ”u

2) H2〇2, NaOH

, -51, (iii)

OH 15 3] 2XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138

(1) Diallyldifluorenylsilane (compound (i)) Dichlorodifluorenylsilane (0.40 in L, 77.5 mm ο 1) was dissolved under distillation in an argon atmosphere. 20 ml of ethyl ether was added dropwise to a 1 M allylmagnesium bromide · ether solution (2 3 2 m 1, 2 2 mm ο 1) under ice cooling, and the mixture was stirred at 50 ° C for 8 hours. After the reaction was completed, 1 N hydrochloric acid (about 150 ml) was added under ice-cooling, extraction was performed with diethyl ether, and the solution was washed with distilled water. The organic layer was dried over anhydrous sodium sulfate, and the solution was concentrated under reduced pressure. The residue was purified by distillation under reduced pressure (54 mm Hg / 58 ° C) to obtain the compound (i) (7.14 g, yield 68.8%) as a liquid. (2) bis (triallylsilyl) difluorenylsilane (compound (ii)) dissolves compound (i) (7.00 g, 49.9 mm ο 1) in an argon atmosphere. To 50 ml of THF after steaming, a catalyst amount of Speier catalyst (0.1M hexahydrate platinum (IV) acid · isopropanol solution in hexahydrate) was added dropwise. Subsequently, triclosan (20.1 ni L, 2 0 ni m ο 1) was added dropwise under ice-cooling, and TH F 3 0 m L was used to wash the solution together with a dropping bucket. After stirring at room temperature for 18.5 hours, the reaction solution was distilled under normal pressure (75 to 80 ° C) to remove the solvent and excess trichlorosilane. To this, add 16 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 Distilled τ HF 60 0 L, dropwise under ice-cooling 1> odorized allyl magnesium · ether solution (6 4 5 m 1,6 4 5 mm ο 1), and the row 0C 1] day to the dish 1.5 hours and 5 (TC 18 hours of stirring. After the reaction is complete, add 1N hydrochloric acid (about 5 0 0 m 1), extracted with ether, and washed the strips with distilled water. The organic layer was dried over anhydrous sodium sulfate, the solution was filtered and concentrated. The residue was subjected to silica gel chromatography (only Purified with hexane) to obtain the compound (ii) (8.33 g '37 .5%) in liquid form (3) bis (tri (3-hydroxypropyl) silylpropyl) difluorenyl (Compound (丨 丨 i)) φ Under an argon atmosphere, 'put 1M BHrTHF complex (30.4mL' 3 0 · 4 mm ο 1) into a flask, and drop cyclohexene ( 2 · 4 8 g, 30. 4 mm ο 1) in THF 50 m L. After stirring, the solution was stirred for 1 hour, and then the compound (丨 丨) (3.00§) was added dropwise under ice cooling. 6 74111111〇1) of it) ^ 9 〇11 ^ solution. After the dropwise addition was complete, 'the reaction solution was stirred at room temperature for 3 hours. Methanol was added dropwise under ice-cooling 20 ml L', and 3.0 · sodium hydroxide (8.99 ml L, 27 · Onimol) 'and 30% hydrogen peroxide solution (9.17mL, 80.9mm〇1). Stir for 1 hour after the end of dropping. Separate the organic layer, extract the aqueous layer with THF and combine with the organic layer. It was washed with saturated brine. After drying, the solvent was distilled off and purified twice by reprecipitation to obtain an oily compound (iii) (3.60 g, 96.7%). Then, the compound ( lll) sulfonium sulfonation to obtain bis [tris (3-monopropyl) lithium propyl] dimethylsilane (compound (1V), 1 DumbbellU) 6 —B ^ 42 () mg ' 5 5.0%). The identification result is not shown. IR (neat) 1239cnr1 (CH2Br) H NMR: (5 (200MHz, CDCh), -0. 〇2 (s, 6H, 2Me), 〇. 62 (m, 20H, 2SiMe2CH2CH2CH2Si, 312XP / Invention Specification (Supplement)) / 94-08 / 941] 2338 17 200538138 6SiCH2CH2CH2Br), 1. 30 (m, 4H, 2SiMe2CH2CH2CH2Si), 1. 81 (m, 12H, 6SiCH2CH2CH2Br), 3. 39 (t, 12H, 6SiCH2CH2CH2Br) 〈with carbon silane branch Synthesis of Halogen Compounds in the Shaped Polymer Framework: Fan (0) 3-Br> is the same as Dumbbe 1 1 (1) 6-B r, and Fan (0) 3-Bro Fan (0) is synthesized according to the following reaction formula. ) 3-Br

IR (neat): 1 2 3 8 c ηΤ 丨 (C Η 2 B r)! Η NMR: (5 (200MHz, CDCl3), 0.95 (m, 6H, SiCH2), 1. 83 (m, 6H, S1CH2CH2) , 3.40 (t, 6H, CH2Br) 〈Synthesis of halogen compounds with carbosilane dendrimer skeleton: Ball (0) 4-Br &gt; Same as Dumbbe 1 1 (1) 6-B r, according to the following Reaction Synthesis Ball (0) 4-Βγο Bal1 (0) 4-Br 18 3] 2XP / Invention Specification (Supplement) / 94_08 / 94] 12338 200538138

IR (neat): 1238cm-丨 (CHhBr) φ! H NMR: 5 (200MHz, CDCl3), 0.70 (m, 2H, SiCH2), 1. 83 (m, 2H, S1CH2CH2), 3.40 (t, 2H, CH2Br ) (Example 1) (Synthesis method) Carbosilane dendrimers carrying α-1,3 Man sugar chains were synthesized in the following order. First, a sugar chain portion was prepared according to the following reaction formula.

<Ethylation of mannose and 1-bromo-4-dagger> Add D-mannose 1 (2.50 g, 1 3 · 9 πι η 1) and acetic anhydride (1 2 ni 1, 19 312XP / Invention Specification (Supplement) / 94-08 / 9411233 8 200538138 1 3 1 mm 0 1), and dripped into a 25% desertified hydrogen-acetic acid solution 4.5 m 1, and was stirred for 12 hours. After confirming the generation of the acetic acid compound, the light was shielded and a 25% hydrogen bromide-acetic acid solution 2 2 m 1 was added dropwise. After stirring for 5 hours, the reaction solution was poured into ice water, and then extracted with CHC 1 3 using a separatory filter. The organic layer was washed twice with water and twice with a saturated NaHC0 3 aqueous solution. Wash once with saturated saline. The organic layer was dried by MgSCh. MgS04 was filtered off, concentrated by an evaporator, and dried by a vacuum pump to obtain compound 2. No further purification was performed for this, and it was used directly as a starting material for subsequent reactions.

<1,2-Ethynylation of Compound 2> Under an argon atmosphere, Compound 2 (22.9 g, 55.6 mmol) was dissolved in 13 ml of acetonitrile, and sodium borohydride (10.5 g, 2 7 8 mm) was added. ο 1), and simmered at room temperature for 2 2 hours. The reaction solution was diluted with ethyl acetate and water, washed once with water and twice with saturated saline. The organic layer was dried over anhydrous magnesium sulfate, and the solution was concentrated under reduced pressure. The compound was purified by Shixi gel column chromatography (hexane: ethyl acetate = 5: 1 ~ 3: 1 ~ 2: 1) to obtain a compound. 3 (yield 11 · 8 g, yield 64%). <Deacetylation and 4,6-benzylidene of compound 3> Under an argon atmosphere, compound 3 (5 · 7 3 g, 17 · 3 mm ο 1) was dissolved in 5.0 ml of methanol, and 曱 was added. Oxy sodium (1 40 mg, 2.6 mm ο 1), and stirred at room temperature for 1 hour. An ion exchange resin I R 1 2 0 B (IT) was added to the reaction solution to neutralize it. After filtering off the resin, the reaction solution was concentrated and dried. It was dissolved in N, N-difluorenylacetic acid amine 1 5 · 0 η〗 1, and phenylpyrene diacetic acid (3 · 7 m 丨, 2 4 · 6 in η 1) was added, followed by addition (+)-10-camphor lutein acid (3 7 9 丨 11 g, 1.6 3 丨 1 m〇1), and stirred under reduced pressure at 30 ° C for 6 hours. After the reaction was completed, the reaction solution was cooled to room temperature. 20 312XP / Invention Specification (Supplement) / 94-08 / 94] 12338 200538138 To this was added triethylamine (0.45 m 1,3. 3 4 mm ο 1), purified by Shixi gel column chromatography (Hengkeng: ethyl acetate S_2 1 ◦: 1 ~ 5: 1 ~ 3: 1 ~ 1: 1) purification to obtain a compound> 4 (yield 5. 0 8 g, quantitative (two-stage)). '<Disaccharide synthesis> Under argon, dissolve compound 2 (298 mg, 0.72 in η) ο 1) and compound 4 (100 mg, 0. 3 4 mm ο 1) Dry inert gas 8.0 in 1. Add dry molecular sieve 4 A powder 1.0 g, and stir at room temperature for 1 hour, followed by stirring at -20 ° C for 1 hour. To the reaction solution was added silver trifluorosulfonate (2 8 mg, φ 0. 8 9 丨 nm〇1), and the mixture was stirred at -2 ° C for 2 hours, and further silver trifluoromethanesulfonate (1 1 3 mg, 0.44 ill mO1), and stirred at -20 ° C for 40 minutes. After the reaction was completed, sodium carbonate (3.02 m g, 2.55 m m ο 1) was added, and then filtered through celite (C e 1 ite). The reaction solution was diluted with aerosol, and washed twice with a saturated aqueous sodium hydrogen carbonate solution and once with a saturated saline solution. After the organic layer was dried over anhydrous magnesium sulfate, the solution was concentrated by filtration. The residue was purified by silica gel column chromatography (toluene: ethyl acetate = 5: 1) to obtain compound 5 (yield: 139 n, yield: 6 6%).

〈Deprotection and Acetylation of Compound 5〉 To Compound 5 (860 mg, 1.3 mg ni ο 1) was added 90% trifluoroacetic acid aqueous solution 10 in 丨, and the mixture was stirred at room temperature for 2 2 hours. . While the reaction vessel was cooled with an ice water bath, sodium carbonate was added to neutralize the reaction solution, and then concentrated and dried. To the residue were added sodium acetate (229 in g, 2.79 丨 τι ni〇1) and acetic anhydride (15 ηι 1, 158 mm ο 1), and the mixture was stirred at 110 ° C for 1 hour. . After the reaction was completed, ice-water was added, extraction was performed by aerosol, and the mixture was washed twice with a saturated sodium bicarbonate aqueous solution and once with a saturated saline solution. The organic layer was dried by anhydrous magnesium sulfate. 21 312XP / Invention Specification (Supplement) / 94-08 / 94] 1233 8 200538138, the solution was passed through; considered concentrated. Purification was performed by Shixi gel column chromatography (hexane: ethyl acetate S_ = 1: 1 to 1: 2) to obtain compound 6 (yield 5 98 mg, yield 64% (two stages)). <Compound 1 of 1-Allylation> Under an argon atmosphere, dissolve compound 6 (4.08 g, 6.0 m in ◦ 1) in dry digas and burn 27 m1, and add dilute propanol (2 m 1, 3 0.7 mm), and cooled to -5 ° C. Boron trifluoride-ether complex (8 m 1,

6 3.1 m m01). After the dripping was completed, the mixture was stirred at 0 ° C for 30 minutes and at room temperature for 7 hours. After the reaction was completed, the reaction solution was poured into ice water, and each was washed once with water, twice with a saturated sodium bicarbonate aqueous solution, and once with a saturated common salt water. After the organic layer was dried over anhydrous osmium sulfate, the solution was concentrated by filtration. The residue was purified by silica gel column chromatography (xylene: ethyl acetate 5: 1 ~ 3: 1 ~ 2: 1 ~ 2: 1 ~ 0: 1) to obtain compound 7 (yield 1.7 3 g, yield 43%). <Sulfurylation of Compound 7> Under an argon atmosphere, Compound 7 (1.73 g, 2.56 mm ο 1) was dissolved in 1,4-diσ, etc., and 1.5 m 1, and Add thioacetic acid (3.7 m 1.5, 2.0 mm ο 1) and heat to 50 ° C. A I Β Ν (2 · 11 g, 12 · 8 m in 〇 1) was added and stirred at 80 ° C for 3 hours. Thereafter, in order to eliminate excess A I B N, cyclohexene (1.5 ml 5 14.8 nim ο 1) was added, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated and purified by Shi Xi gel column chromatography (xylene: ethyl acetate S purpose = 10: 1 ~ 5: 1 ~ 3: 1 ~ 2: 1), followed by Sephade XL Η -2 0 (developed with methanol) to obtain compound 8 (yield 1.87 g, yield 97.7%). According to the following reaction formula, the sugar chain-carbosilane dendrimer bone 22 312XP / Invention Specification (Supplement) / 94-08 / 94] 12338 200538138 introduction reaction and deprotection were performed to prepare F a η (0 ) 3-α '-1, 3-M a η.

Fan (0) 3-Br 10 / η9οη 11 HO 〇Η

Fan (0) 3-a-1,3-Man <Introduction reaction of sugar chain to carbosilane dendrimer skeleton>

Under an argon atmosphere, the compound 8 (389 mg, 0.5 2 mm ο 1) and the halogen compound 9 (Fan (0) -Br: 40.2 mg, 8 5 .3 mm ο 1) It was dissolved in N, N-diamidinofluorenamine 0.4 Hi 1, and 0.4 ml of methanol was added to the mixture, followed by stirring at room temperature for 1 hour. To this was added sodium methoxylate (30.1 mg, 0.56 m m ο 1), and the mixture was stirred at room temperature overnight. After the reaction was completed, 0.5 ml of acetic acid was added, and after stirring at room temperature for 10 minutes, it was concentrated, and the residue was suspended in 1.0 ml of pyridine, and then acetic anhydride (2.0 m 1 · 0 mm ο 1) Stir overnight at room temperature. After the reaction solution was concentrated, ice water was added, and the mixture was extracted with aerosol. The organic layers were washed once with 1 N hydrochloric acid, the strips were washed twice with a saturated sodium bicarbonate aqueous solution, and once with a saturated saline solution. After the organic layer was dried over anhydrous magnesium sulfate, the solution was concentrated by filtration. The residue was purified by silica gel column chromatography (Hexion: ethyl acetate, 1: 2 ~ 1: 2 ~ 0: 1) and GPC to obtain F a η (0) 3-a-1, 3-M a η (0 A c) 1 0 (yield 6 1.2 mg, yield 30% (two stages)). The identification results of the compounds are shown below. 13 CNMR (CDC 1 3) d (pp in); 9 7. 3 (C-1), 9 8.9 (C-Γ). HRMS (ES I): CmHiuOsdsS i Na [M + Na Γ calculated 2 3 8 1 · 7 5 0 8, 23 312XP / Invention Specification (Supplement) / 94-08 / 94] 12338 200538138 Found 2 3 8 1. 7 4 8 5. [A] u (3 2). + 3 2. 3 1 (c = 1 · 0 1 n CHC 1 3). The same applies to Β a 1 1 (〇) 4-a-Β r, Du u η bbe 1 1 (1) 6-a-Β r The reaction was carried out on the ground, and a carbonaceous yakana dendrimer carrying a sugar chain protected by acetamidine in a sugar portion was synthesized separately. The identification results of each compound are shown below. B a 1 1 (0) 4-a, 1,3-M a η (0 A c): Yield 8 1.1 .1 g (yield 35% (two stages)) 13 CNMR (CDC 1 3) d (ppm) 9 7. 3 (C-1), 9 8. 8 (C-Γ). HRMS (FAB): C &quot; 8H &quot; (″ 〇72S ^ iSi [Μ + ΗΓ Calculated value 3033. 9780, Measured value 3 0 3 3 · 9 7 5 1 · [a] d (), + 3 3. 8 6 (c: 1.0 in CHC 1 3) ·

〇111111 ^ 611 (1) 6-3-1, 3-1 ^ 11 (0.8 (:): yield 61.31 ^ (yield 31% (two stages)) 13C NMR (CDClOd (ppm) 97.4 (Cl ), 98.9 (01 ') · HRMSCFABkCzoHmOwSGSiJM + Hr calculated 4706. 5693, found 4 7 ◦ 6. 5 6 7 9 · [a] iu 3 3), + 3 3 · 0 0 (c = 1. 0 in CHC 13) <Deprotection of carbosilane dendrimers with sugar chains> Fan (0) 3-al, 3-correction 3: 1 (080〇) 10 (58.51 ^ '24 .8 丨 11 丨 11 〇1) Dissolved in acetol 0.3 m 1, added acetic acid sodium alcohol solution (1.0 M, 70 m 1 ′ 70 mm ο 1), and stirred at room temperature for 1 hour, then added 〇 1M aqueous sodium hydroxide solution, and stirred overnight at room temperature. After adding acetic acid to neutralize the solution, it was concentrated and subjected to gel filtration to remove the inorganic salt to obtain the target compound. 8 11 (0) 3- 〇: -1, 3-1 ^ 131111 (yield 44.61) 1. The compound identification results are shown below. 13 CNMR (D 2 0) d (ppm); 9 9. 6 (C-1), 1 0 2 (C-Γ). HRMS (FAB) ·· C g ϋ Η! Ϋ 'I 0 3 3 S 3 S i N a [Μ + N a Γ Calculated value 1 4 9 9 · 5 2 8 9 Found 1 4 9 9 · 5 2 7 8. [A] w 2 2), + 7 8. 7 1 (c = 0 · 8 7 i η Η 2 0). About having B a 1 1 (0) 4 Type skeleton, Dumbbe 1 1 (1) Type 6 skeleton load 24 312XP / Invention Specification (Supplement) / 94-08 / 94] 12338 200538138 Carbon stone with sugar bond Deprotection should be carried out on the ground, and the target carbosilane dendrimers carrying sugar chains were synthesized separately. The identification results of each compound are shown below.

Ball (0) 4-a -1,3-Man: Yield 75.5nig (quantitative) 13 CNMR (D 2 0) d (ppm); 9 9 0 (C-1), 1 0 1.4 (C -Γ). HRMS (ESI): C 7 2 Η! 3 2 0 “SS i N a [Μ + N a]. Calculated value 1 8 7 9. 6 6 4 1 Measured value 1 8 7 9. 6 6 2 2 · [a] d (m + 1 0 0 · 4 1 (c = 1 · 0 i η H 2 0). 〇111] 1135611 (1) 6-〇: -1, 3-1 ^ 11: Yield 33.8111 Avoid (yield 90%)

Anti-defense bureau points H2〇). 13 CNMR (D 2 0) d (ppm); 1 0. 7 (C-1), 10 3. 1 (C-Γ). HRMSCESIhCwihHOwSGSisNaz / ^ fM + ZNarVZ calculated value 1 4 9 3. 5 4 8 7, measured value 1 4 9 3. 5482. [a] d⑴), +48. 2 5 (c = 1. 0 in (Example 2) First, prepare according to the following reaction formula Sugar chain part.

Sodium acetate (2.51 g, 30. 6 in m ο 1) was suspended in acetic anhydride (25.0 m 1, 263 m ηι1), and heated to 110 ° C. Add D-mannose 1 (5.00 g? 2 7. 8 η η ο 1) in small amounts and stir for 2 hours. After the reaction was completed, ice water was added. Extraction was carried out with gas imitation, and the county was washed twice with saturated sodium bicarbonate water and once with saturated brine. After the organic layer was dried with anhydrous money, the solution was concentrated and concentrated. This was placed in a two-necked flask and replaced with argon. To this was added dry dioxane 1 2 3 m 1, allyl alcohol 3] 2XP / Invention Specification (Supplement) / 94-08 / 94] 12338 sugar, solution sulfuric acid environment 25 200538138 (9.5 m 1, 1 3 9 in 丨] 10) and cooled to -5 ° C. The trifluoride pentamethylene-acetate complex (9 4 m 1, 7 4 1 m m ο 1) was added dropwise over a period of 30 minutes. After the addition was completed, stirring was performed at 0 ° C for 30 minutes and room temperature for 5 to 4 hours. After the reaction was completed, the reaction solution was poured into ice water, and the strips were washed once with water, twice with a saturated sodium bicarbonate aqueous solution, and once with a saturated saline solution. After the organic layer was dried over anhydrous magnesium sulfate, the solution was concentrated by filtration. The residue was purified by silica gel column chromatography (toluene: ethyl acetate = 5: 1) to obtain compound 12 (yield 7.53 g, yield 70% (two stages)).

<Sulfurylation of Compound 1 2> Under an argon atmosphere, Compound 1 2 (3.65 g, 9. · 40 m πί ο 1) was dissolved in 1,4-diisobutane (2 · 0 m 1), thioacetic acid (1 3 · 4 m 1 '1 8 8 mm ο 1) was added thereto, and heated to 50 ° C. To this was added A I B N (7 · 7 2 g, 4 7.0 mm m ο 1), and stirred at 80 ° C for 2.5 hours. Thereafter, in order to eliminate excess A I B N, cyclohexane was diluted (5.0 in 1, 49.3 m m ο 1), and stirred at room temperature for 30 minutes. The reaction solution was concentrated and purified by silica gel column chromatography (xylene: ethyl acetate = 1 0: 1 ~ 5: 1 ~ 3: 1) to obtain compound 1 3 (yield 3.16 g, Yield: 73%). In the same manner as in Example 1, the reaction of introducing sugar bonds into the dendritic south molecular skeleton of the carbon stone pit was performed. The identification results of each compound are shown below. Fan (0) 3-Man (0Ac): Yield 107.9mg (yield 66% (two stages)) 13C NMR (CDCl3): d (ppm); 97.4 (C-1). HRMS (ESI): C 6 ϋ Η 9 8 0 3 ϋ S 3 S i N a, [Μ + N a Γ Calculated value 1 5 1 7. 4 9 7 2, Measured value 1 5 1 7 · 4 9 9 0 · [a] m '2 9 ), + 4 2. 3 (c 1 1.0 in CHC 1 〇 · B a 丨 1 (◦) 4-M a η (0 A c): Yield 1 2 0. 3 mg (yield 6 6% ( Two stages)) 13 CNMR (CDC 1 3): d (pp in); 97.5 (C-1). 26 3] 2XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 HRMS (FAB) : C 8 ο Η 12 5 ◦ 4 ϋ S 4 S i, [Μ + Η] * Calculated value 1 8 8 1. 6 3 9 9, Measured value 1 8 8 1 · 6 4 4 5. [A] i) (2 9), + 4 5. 1 (c = 1 · 〇in CHC 1 3).

Du〇] bbell (l) 6-Man (OAc): Yield i4i.6mg (62% yield (two-stage)) 13C NMR (CDCl3): d (ppm) 97.4 (Cl). HRMS (FAB): Ci28H2U5 ( K〇S (5Si3 [M + H] -calculated value 2978.0622, actual value 2 9 7 · 0 6 6 9. [A] d (2 9). + 4 〇. I (c = i. 〇 丨 n CHC 1 3) The deprotection was performed in the same manner as in Example 1. The identification results of each compound are shown below.

? &amp; 11 (0) 3-1 ^ 11: Yield 54.811 ^ (yield 61%) 13C NMR (D2〇) d (ppm); 100.7 (C-1). HRMS (ESI): C42H74〇18S3SiNa, [ M + Na] + calculated 1013.3704, found 1013.3696. [A] iu27), +48.96 (c = 1.0 in H2〇).

Ball (0) 4-Man: Yield 64.8mg (yield 82%) 丨 3C NMR (D2〇) d (ppm); 99.9 (C-1). HRMSCESIkCuHnChdS ^ SiNajM + Nar Calculated value 1231.4 528, Found 1 2 3 1 · 4 5 8 1. [A] d (2 4). + 5 2. 7 (c = 1. 0 i η H 2 0). 〇1111113 乜 611 (1) 6-1 ^ 11: Yield 33.611 ^ (Yield 81%) 13C NMR (D2〇) d (ppm); 100 (C ~ 1). HRMS (FAB): C8〇Hi56〇3GS6Si3Na, [M + N a] Calculated value 1991.7906 'Found 1 9 9 1. 7 9 3 7. [A] d (3 Q), + 4 6 · 3 (c = 1.0 0 η H 2 0) · (Example 3) First, prepare a sugar chain according to the following reaction formula Receptor. 27 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138

NaOCHa {CH3) 2C {OCH3) 2 CH3OH CSA, Drierlt © yield 60% (two-stage) 4-pinene small alcohol, BF3, OEt2 CH2CI2 yield 40%

Ac0l / 0Ac〇 OAc

AcOy- ^ r-c OAc c〇Ac 15

BzCl pyridine

CH2CI2 50% CF3C〇〇Haq.

Yield 95% Yield 86% 18

</ 3 of lactose Selective complete acetylation: 0-(2,3,4,6 -tetra-0 -acetamido- / 9 -D -galactopyranoside)-(1-4)- Synthesis of 1,2,3,6-tetra-O-ethynyl_ / 3-D-grapepiperanoside (1 4)> Add acetic anhydride (5001011, 5.311101) to sodium acetate (47.9 Avoid (584111111〇1) and let it suspend, the temperature of the suspension reaches 110t under stirring. Secondly, add lactose-hydrate (10 5. 3 g? 2 9 2 ni m ο 1) in small amounts while paying attention to temperature increase: mix for 2 hours. After the reaction was completed, the reaction solution was added to excess ice water to hydrolyze unreacted acetic anhydride. The precipitate was taken out, washed with water, and then recrystallized by ethanol to obtain crystals of fully ethylated lactose acetate 14 (14.9.1 g, 75.3%).

<Introduction of glycosphingyl base: dilute group 0- (2,3,4,6_tetra_0_ethydonyl-yS -D-galactopyranosyl)-(1- &gt; 4) -2 Synthesis of 1,3,6-triacetoxy-cold-D-glucopyranoside (1 5)> Under argon atmosphere, make lactose acetate 1 4 (50. 0 g, 7 3. 7 in m ο 1) Dissolved in 3 m 2 methane, added 4-pentene_ 1-ol (3 8 0 m 1, 0.3 7 ηι 0 1), and cooled to -1 1 ° C . In response to this, while paying attention to the temperature rise, a boron trifluoride-ether complex (9 3.4 in 1, 0.7 4 in 0 1) was dripped into the mouth at a time of about 1 hour. After stirring at 0 ° C for 1 hour, the temperature was returned to room temperature and stirring was continued for 28 312XP / Invention Specification (Supplement) / 94-08 / 9411233 8 200538138 for 3 hours. After the reaction was completed, the reaction solution was poured into ice water and extracted with chloroform, and the organic layers were sequentially washed with water, a saturated aqueous sodium hydrogen carbonate solution, and a saturated saline solution. After the organic layer was dried with anhydrous sulfuric acid and concentrated, the mixture was purified by silica gel column chromatography (developing system: toluene, and then toluene: ethyl acetate = 2: 1) to obtain compound 1 5 (2 1. 3 g, 4 1% ° <de- 0 _ Ethylene: 0- (/ 3-D -galactopyranosyl)-(1 — 4) -stone-D-grape Synthesis of Yumansu bitter>

Under argon atmosphere, compound 15 (2 1.2 g, 30.1 mm ο 1) was dissolved in dry methanol 200 m 1, and sodium methoxylate (1.14 g, 2 1 · 1 inm ο 1), and stirred at room temperature for 4 hours. After the reaction was completed, a cation exchange resin (Amberlite IR120B manufactured by ORGANO) was added to neutralize the reaction solution. After removing the resin by filtration, the filtrate was concentrated to obtain Compound 3.7 g. No further purification was performed for this, and it was used directly in the next reaction. ) <Isopropylidene: Pentenyl 0- (3, 4-0-isopropylidene- / 3-D-galactoperanose) _ (1-4) -yS -D_ grape slightly ran Synthesis of sugar bitter (16)> The above compound (8.68 g, 2 1 · 15 mm ο 1) was dissolved in dry DMF (1 8 0 m 1) under an argon atmosphere, and acetone di was added thereto. Amidino acetal (5.18 m 1, 4 2. 2 9 m η 1) and Drieri 1: e 9. 0 g. After stirring at room temperature for 30 minutes, C S A (0.49 g, 2.12 ηιηο 1) was added, and stirred at room temperature for 1 hour, and then heated to 80 ° C and stirred for 4 hours. After the reaction was completed, the reaction solution was cooled using an ice bath, and sodium bicarbonate was added to neutralize it. The suspension was filtered through celite and the filtrate was concentrated. The residue was purified by silica gel column chromatography (developing system: chloroform: methanol 2 10: 1) to obtain isopropylidene 29 at the 3 ′ and 4 ′ positions. 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 Protected compounds 16 (5, 6 9 g, 59%). <Synthesis of diols: Pentenyl 0-(2,6-di-0 -phenylfluorenyl-/ 3-D -galactosigma sulose)-(1-4)-(2, 3, Synthesis of 6-Tri-O-Benzyl Alkyl- / 3-D-Grape Branose (1 8)>

Under an argon atmosphere, the isopropylidene 16 (5. 13 g, 11.39 mmol) was dissolved in dry pyridine 5 5.0 m 1 and cooled to 0 ° C. Add benzamidine gas (78.7 m 1, 0.6 m) at 0 ° C, return to room temperature, and stir for 3 hours. After the reaction is completed, water is added to neutralize excess radon compounds. The reaction solution was extracted with chloroform, and the organic layer was sequentially washed with 1 N hydrochloric acid, a saturated aqueous sodium hydrogen carbonate solution, and a saturated saline solution. The organic layer was dried over anhydrous magnesium sulfate and concentrated. Compound 17 was thus obtained. The 24.0 g of the residue obtained from the above reaction was dissolved in dichloromethane 1 10 in 1 and then water 2 5 m 1 was added. The mixed solution was cooled to 0 ° C, and trifluoroacetic acid (2.5 ml) was added dropwise, followed by stirring at room temperature for 3 hours. After the reaction was completed, the reaction solution was washed with water, a saturated sodium bicarbonate aqueous solution, and a saturated saline solution in this order, and the organic layer was dried over anhydrous magnesium sulfate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (developing system was hexane: ethyl acetate 2M and 1: 1) to obtain a sugar chain acceptor 1 8 (8.15 g, yield 77.7%). Next, a sugar chain donor was prepared according to the following reaction formula.

30 312XP / Invention Specification (Supplement) / 94-08 / 94 Π 2338 200538138

Agricultural rate 45% 22 I rate 53% 23

Yield 98% (two stages) 26 H2, Pd cat. EtOH, EtOAc 89%

27

Vilsmeier reagent 2,4,6-Colin Mi CHaCIa yield 72%

AcO

AcO

28 &lt; &gt; Odorization (synthesis of 2 lactose and bitterness: Odoration 0 _ (2,3,4,6 -tetra-0-ethanyl-yS-galactopyranosyl)-(l-4)- Synthesis of 2,3,6-tris-0-acetamido-a-D-glucomannan (1 9)> Make lactose acetate 1 4 (6 0. 0 g, 8 8. 4 mm ο 1 ) Dissolved in 300 0 in 1 acetic acid, add hydrogen bromide (30% acetic acid solution) (5 3 · 0 in 1, 0 · 2 7 m ο 1), and seal it. Stir at room temperature with light shielding 3.5 hours later, the reaction solution was poured into ice water and extracted with chloroform, followed by washing with water, a saturated aqueous sodium bicarbonate solution, and a saturated saline solution in sequence. The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to hot filtration using chloroform 100 m 1. After the solution was cooled to room temperature, diethyl ether 2 00 in 1 was added to perform crystallization to obtain a needle-like compound 1 9 (5 3 · 1 g, 86%). <Formation of aldose lactone (1 acta 1): 0- (2,3,4,6-tetra-0-acetamido-cold-D-galactopyranoside Sugar)-(1-4) -3,4,6-tri-0-Ethyl-D-enose (2 0) 312XP / Invention Specification (Supplement) / 94-08 / 94 2338 31 Synthesis of 200538138> Zinc powder (4 5 · 7 g, 0.69 9 m ni 〇1), sodium acetate (86.0 g, 1.05 η 1), and copper sulfate pentahydrate (4.36 g, 17.5 mm ο 丨) were suspended in water 2000 m 1. Compound 1 9 (4 8 · 9 g, 6 9. 9 mm ο 1) dissolved in acetic acid

3 0 0 丨]] 1, and this solution was dropped into the ice-cold suspension for about 1 hour. The reaction solution was allowed to warm up naturally from 0 ° C and stirred for 5 hours. After the reaction was completed, the zinc powder was removed by filtration, and the mixture was extracted with aerosol, followed by washing with water, a saturated aqueous sodium hydrogen carbonate solution, and a saturated saline solution in this order. The organic layer was dried over anhydrous magnesium sulfate, and then concentrated. The residue was purified by Shi Xi gel column chromatography (developing system: toluene: ethyl acetate = 2: 1) to obtain aldose lactone 20 (31. 4 g, 8 ◦%). <Azide nitrification> Under an argon atmosphere, CAN (17.6g, 32.1mmol) and sodium azide (1.04 g, 16.1 mm ο 1) were dried under reduced pressure for 5 hours, and then Cool to -2 ° C. Aldulactone 20 (6.0 g, 10.7 m η 1) dissolved in dry acetonitrile 5 4 hi 1 was added to the dry powder. After stirring at -2 ° C overnight, the suspension was poured into ice water, extracted with ethyl acetate, and the bars were washed sequentially with water and saturated saline. The organic layer was dried over anhydrous magnesium sulfate, and then concentrated. The residue was separated by silica gel column chromatography (developing system: toluene: ethyl acetate 2: 1) at two main positions (s ρ 〇 t), and the residue containing the isomers was separated. The mixture gave compound 21 (5.11 g, 71%). &lt; Alpha bromide: Brominated 0-(2,3,4,6 -tetra-0 -Ethyl- / 3-D -galactoperanose)-(1-4)-3,6 -di Synthesis of -〇-acetamidine-2-azide-2-deoxy-α-D-glucopyranose (2 2)> 32 312 XP / Specification of the Invention (Supplement) / 94-08 / 94112338 200538138 Under Ar Under atmospheric conditions, the compound 2 1 (17.7 g, 2 6 · 7 mm ο 1) and brominated chloride (1 1 · 6 g, 0.1 3 4 m ο 1) were suspended in dry acetonitrile 1 8 0 m 1, and stirred at room temperature for 4 hours. The reaction solution was diluted with ethyl acetate, and washing was performed sequentially using water and saturated saline. The organic layer was dried over anhydrous magnesium sulfate, and the solution was concentrated, and then roughly purified by a short column, followed by (ethyl acetate-hexane), to obtain a compound 2 2 (8.2 g, 4 5%).

&lt; / 3 -benzyl: benzyl 0-(2, 3, 4, 6-tetra- 0-acetamido-dot-D-galactopranose)-(1-4) -3,6_ Synthesis of Di-O-Ethyl-2-azide-2-deoxy- / 3-D-Grape Sulfate (2 3)> Under argon atmosphere, shading was performed to make benzyl alcohol (2 · 1 m 1, 2 0.6 mm ο 1), carbonate (6.6 3 g, 24. 03 m mol) 'and D rierite (5 g) suspended in dry nitric methane 20 m 1. The suspension was cooled to -2 ° C, and a solution of compound 2 2 (4.69 g 5 6 · 8 7 him ο 1) in a 30 in 1 solution was added dropwise. After the addition was complete, stir overnight at -20 ° C. The reaction mixture was filtered through celite, and the solution was diluted with ethyl acetate. The organic layer was washed with water, a saturated sodium bicarbonate aqueous solution, and a saturated saline solution in order, and dried over anhydrous sulfuric acid. After concentration, it was roughly purified by a short column and recrystallized with 15 ml of acetone to obtain compound 2 3 (2.75 g, yield 56.6%) as white crystals. <De-O-ethyl fermentation: nodyl 0_ (yS-D-galactobranose)-(1-4)-2-azide-2-deoxy-cold-D-grape piperanoside (2 4 ) Synthesis> Under argon, dissolve compound 2 3 (4.27 g, 6.01 mm ο 1) in dry methanol 4 0 丨 11 1 and add sodium ethoxide (19.5 in g , 3 · 6 1 m in ο 1), and stirred overnight at room temperature. The precipitated crystals were decanted to obtain compound 2 4 (2.62 g, 95 ° / 〇). This was not purified, and was used directly in the next reaction. 33 312XP / Invention (Supplement) / 94-08 / 94112338 200538138 <Reduction of azido: benzyl 0-(/ 3-D -galactopyranosyl)-(1-4) 2 -amino- Synthesis of 2-deoxy- / 3-D-grapepiperanoside (2 5)> Compound 2 4 (1 _ 8 3 g, 4. 0 4 in in ο 1) was dissolved in a dry bite 50 m 1. To this, triethylamine 15 m 1 was added. The reaction solution was aerated with hydrogen sulfide gas for 1 hour, and then the reaction system was sealed and left overnight. The reaction solution was concentrated, and the residue was extracted with a mixed solution of water: methanol = 1: 1. The extract was filtered through diatomaceous earth, and the filtrate was concentrated to obtain compound 25 (1.72 g, 99%). This was not purified, and was used directly in the next reaction.

<Phthalidene: benzyl 0- (2,3,4,6-tetra-0-ethylfluorenyl-cold-D-galactoperanose)-(1-4) -3,6-di-〇 -Acetyl-2_deoxy-2-phthalimidine- / 3-D-Synthesis of glucopyranoside (2 6)> Under argon atmosphere, make compound 2 5 (1 · 7 2 g , 4 · 0 0 mm ο 1) soluble in dry ° specific bite, add S too anhydride (3 5 5 · 2 mg, 2. 40 mm ο 1), and stir at 70 ° C for 3 5 minutes . Then add triethylamine (560 m 1, 4. 00 m ο 1) and hydrazone anhydride (35.5.2 mg, 2.40 mm〇1), and stir at 70 ° C for 4 hour. After cooling to room temperature, 10 ml of methanol was added, and after stirring for several minutes, the solvent was distilled off under reduced pressure. Suspend the residue in a dry ° specific bite (2 3.2 m 1, 2 8 8 mm ο 1), add ethyl δ dry (1 3 · 6 丨 τι 1, 1 4 4 m in ο 1), Mix at 90 ° C for 2 hours. After the reaction was completed, the reaction solution was concentrated, and the reaction mixture was purified by silica gel column chromatography (developing system was hexane · ethyl acetate = 1: 2) to obtain compound 2 6 (3 · 19 g, yield 98%). <Disassociation: 〇_ (2,3,4,6-tetra-0-ethynyl- / 3-D-galactopyranosyl)-(1-4) -3,6-di-〇- Synthesis of Ethyl-2-deoxy-2-phthalimido-imine-α, dot-D-grapepiperanose (2 7)> 34 312 XP / Invention Specification (Supplement) / 94-08 / 94] ] 2338 200538138 The compound 2 6 (3, 19 g, 3. 9 2 n] m ο 1) was dissolved in ethyl acetate: ethanol = 1: 1 solution 50 0 π 丨, 20% atmospheric oxidation was added / activity 1.5 g of carbon was vigorously stirred in a hydrogen-oxygen environment using a contact reduction device. After confirming the completion of the reaction by TLC, the reaction solution was filtered through celite. After the filtrate was concentrated, the residue was purified by silica gel column chromatography (developing system: toluene: ethyl acetate = 1: 1 and 1: 2) to obtain compound 2 7 (2.53 g, yield). 89%).

&lt; Cold gasification: 0-(2,3,4,6_tetra-0 -acetate-yS -D -galactopyranosyl)-(1-4) -3, 6_di-0_ Synthesis of ethenyl-2_deoxy-2-phthalimidoimine- / 3-D-grapepiperanose (2 8)> Under an argon atmosphere, the Vi i 1 smeier reagent (3 3 6. 4 mg, 2 · 6 3 m in ◦ 1) suspended in 2 m 1 of dry dioxane. In addition, compound 2 7 (6 34.0 mg, 0.876 丨 11111〇1) and 2,4,6-Collin test (17〇1111,1.3111111] 〇1) were dissolved in dry dichloromethane 4 in 1 This solution was dropped into a suspension cooled to 0 ° C. After the dropwise addition was completed, the mixture was stirred at room temperature for 3 hours. The reaction solution was diluted with toluene, and then extracted with toluene, and washed sequentially with water, 1 M hydrochloric acid, a saturated sodium hydrogen peroxide solution, and a saturated saline solution. The organic layer was dried over anhydrous magnesium sulfate and concentrated. Organic layer. Dissolve the residue in 3 ml of dioxane and add 5 ml of diethyl ether to perform crystallization. By separating the crystals, a glucose-based donor 2 8 (4 69.1 mg, yield The rate is 72%). According to the following reaction formula, glycosylation of a sugar chain donor and a sugar chain acceptor is performed to prepare a sulfur compound.

312XP / Invention (Supplement) / 94-08 / 9411233 8 35 200538138 n-BuNH2 Ac20 CH3OH Pyridine Yield 79% AcSH, AIBN 1,4-Secondary pit yield 98%

OAc? AJ〇Ac AcO- AcO AcO

OAc NHAc OAc OAc

AcO Acor ^ o ^ o ^ 〇Γ ^ {AcO 〇Ac NHAc OAc ^^ OAc 31 SAc 〈Sugar bittering: pentyl 0- (2,3,4,6_tetra-0-ethyl brew- / 3- D-galactolum kounan • sugar)-(1-4) -0- (3,6-di-0-ethylfluorenyl-2-deoxy-2-phthalimide- / 3-D- Grape σ melon σ south sugar)-(l- &gt; 3) -0- (2,6-di-0-phenylpyrene- / 3-D-galacto-0 citrulose)-(l-4) Synthesis of -2,3,6-tris (phenylhydrazone) -cold-D-glucopyranoside (2 9)> Under an argon atmosphere, the sugar chain donor 2 8 (1.66 g, 2. 2 3 mm ο 1) and sugar chain acceptor 18 (2.50 g, 2. 68 mm ο 1) were dissolved in dry nitroprane 50 m 1 'with M s 4 A powder 5. 0 g, and stirred at room temperature for 2.5 hours to allow the system to dry sufficiently. After shading, cooling the reaction solution to 0 ° C, adding three

Silver fluorosulfonate (2 · 8 7 g, 1 · 1 7 min in ο 1). After stirring at 0 ° C for 2.5 hours, since the reaction stopped, 2.87 g of silver trifluorophosphonium sulfonate was added again, and the mixture was stirred overnight at room temperature. The suspension was filtered through diatomaceous earth, the filtrate was diluted with ethyl acetate, and the organic layer was washed with water, a saturated aqueous sodium hydrogen carbonate solution, and a saturated saline solution in this order. The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (developing system: hexane: ethyl acetate = 1: 2) to obtain a tetrasaccharide 2 9 (2.78 g, 7 6%). 312XP / Invention Specification (Supplement) / 94-08 / 941] 2338 36 200538138 <Transformation of Protecting Group: Pentenyl 0-(2, 3, 4, 6-Tetra-◦-Ethyl- / 3-D -Galactopyranosyl)-(1-4) -〇- (3,6-di-〇-ethyl SE-2-ethyl ethylamino-2 -deoxy_D-grape glucomannan)- (1-3) -0-(2,4,6-di-O-ethyl-derivative- / 3 galactomannanose) _ (1-4) _2,3,6_tri-acetate-grapepiper Synthesis of Glucoside (30)

Under an argon atmosphere, Compound 2 9 (30.3.1 mg, 0.18 5 mm 1) was added with dry methanol 5 ml 1 and n-butylamine 5 μm 1 and heated under reflux for 2 4 hours. The reaction solution was concentrated, and the residue was suspended at a dry ratio. To 5 m 1, 2.5 m 1 of acetic anhydride was added thereto, and the mixture was stirred at room temperature for 2 days. The reaction solution was poured into ice water to hydrolyze acetic anhydride, and then extracted with ethyl acetate. The organic layer was sequentially washed with 1 M hydrochloric acid, a saturated aqueous sodium hydrogen carbonate solution, and a saturated saline solution, and then dried by anhydrous thorium sulfate. . The organic layer was concentrated, and the residue was purified by flash silica gel column chromatography (developing system: hexane: ethyl acetate = 1: 9) to obtain compound 3 0 (1 8 7. 5 mg, 7 9 %). <Thioacetic acid: w -thioacetate-pentanyl 0-(2, 3, 4, 6 _tetra-0 _ acetate-stone-D-galactopyranosyl)-(1-4) -0- (3,6-di-0-acetic acid-2-ethynamido-2-deoxy-D-glucomannanose)-(1-3) -0- (2,4,6-di- Synthesis of 0_Acetyl- / 3-D-galactopyranosyl)-(1-4) -2,3,6-tri-ethyl SE-cold-D-grape piperanoside (3 1)> Under argon atmosphere, compound 3 0 (8 9 6 · 3 mg, 0.7 0 0 in in ο 1) was added with 1, 4-two-port isothermal (0 · 5 m 1) and thioacetic acid (9 9 0 m 1,1 4. 0 mm ο 1). A I B N (29.8%, 1.4 ni ni 1) was added, and the mixture was heated steadily, and stirred at 80 ° C for 2 hours. Then return to room temperature, add cyclohexene (1.4 2 m 1, 1. 40 in in ο 1), good: mix for a few minutes. The reaction solution was roughly purified by a short column, 37 312XP / Invention Specification (Supplement) / 94-08 / 94 2338 200538138, and then purified by GPC (column 1 Η · 2 Η) to obtain thioethyl carbohydrate 3 1 (9 3 1. 0 mg, 9 8%) ° According to the following reaction formula, the introduction reaction of carbon chain dendritic skeleton and the deprotection of sugar chain are carried out to prepare F a η (0) 3 -pair Erythrocyte glycoside ester-〇Η33 °

〈Introduction of erythrocyte glycoside ester derivative to carbosilane dendrimer skeleton: Synthesis of Fan (0) 3-paragloboside-0Ac (32)〉 Under the argon environment, the terminal molybdenum carbon was burnt and dendrimers Plant 811 (0) 3-81_ (25.0 丨 1 ^, 0.053111 丨 11〇1) and tetrasaccharide thioethyl brewer 3 1 (3 0 1.2 mg, 0.2 2 2 mm 0 1) were dissolved in dry DMF After 0.3 m 1, dry methanol 0.3 m 1 was added and thoroughly stirred. The previously prepared sodium alkoxide (1 M methanol solution) 2 4 4 m 1 was added dropwise, and the mixture was stirred at room temperature for 1 hour. 0.2 ml of acetic acid was added, and after neutralization with sodium methoxylate, the suspension was concentrated. To the reaction mixture, 4 ml of dry pyridine and 2 ml of acetic anhydride were added, and the mixture was stirred at 30 ° C for 3 hours to carry out acetylation. The reaction solution was poured into ice water to hydrolyze acetic anhydride, and then extracted with ethyl acetate, and the organic layer was sequentially washed with 1 M hydrochloric acid, a saturated aqueous sodium hydrogen carbonate solution, and a saturated saline solution, and anhydrous sulphate was used. dry. 38 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 The organic layer was concentrated, and the residue was purified by GPC (column 2 Η · 2.5 Η) to obtain a protected acetic acid carrier. Fan type carbosilane dendrimer 3 2 (142. 0 mg, 64%) of erythrocyte glycoside ester derivative. When measuring the molecular weight, the molecular peak (p a r en t p e a k) has a weak intensity. Therefore, a high resolution energy measurement is performed based on the peak value of the divalent ion.

<De-O-ethylation: Synthesis of Fan (0) 3-paragl〇b〇side-OH (33)> Under an argon atmosphere, the acetonitrile 3 2 (109.5 mg, 0. 0 2 6 mm ο 1) dissolved in dry methanol (1 · 0 m 1). To this was added fluorinated sodium (5 · 5 mg, 0.10 mm ο 1). After stirring at room temperature for 30 minutes, 5.0 ml of a 0.1 M sodium hydroxide aqueous solution was added, and stirring was continued. After stirring for 2.5 hours, add 0.1 M sodium hydroxide aqueous solution 5.0 ml and stir; stir for 2.5 hours. After the reaction was completed, a cation exchange resin (Ambeer 1 Iter I R 1 2 0 B manufactured by 0 R G A N 0) was added to neutralize the solution. Filter through a cotton plug to remove resin and concentrate the mash. The residue was purified by gel filtration (G 50, 5% aqueous acetic acid solution) and freeze-dried to obtain an unprotected compound 3 3 (7 1.2 in g, quantitative). The identification results are shown below. NMR: 'H (D2〇, 400MHz): d (ppm) 7.31, 7.15 (each br s, 5H,

SiPh), 4. 26-4. 34 (m), 4. 01 (br s), 3. 75-3.82 (丨 n), 3.58 -3.72 (m), 3. 37-3. 54 (m), 3. 17 (brm), 2.32 (br s, 1 2H, -CH2S), 1.90 (s, 9H, NHAc), 1.41 (br s), 1.25 (br s), 0. 4 9 (brs, 6 H, S i CH 2-). NMR: 13C (D20, 100MHz): d (ppm); 181.09, 134.03, 128.05, 102. 93, 102. 73, 102.39, 82.24, 78.48, 78.27, 75.34, 7 4 .8 9, 7 4. 5 6, 7 2. 9 0, 7 2.5 3, 7 2.2 2, 7 0.9 9, 70. 21, 39 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 6 9,9 4, 6 8.5 8, 6 8.2 9, 6 1. 0 5, 6 0.2 7, 6 0. 1 1, 5 9.8 9, 55.25, 35. 50, 31.68, 29. 39, 28.95, 25.00, 23.92, 22.45, 11.72 ESI-MS Anal. Cni8Hl85N3〇63S3Si [M + Na] ': Calculated 2 6 7 9. 0 1 9 4; Found 2 6 7 9. 0 0 9 6. Prepare Ball (0 ) 4_paragloboside_0H (35).

65% yield

&lt; Synthesis of Ball (0) 4-paragloboside-OAc (34)> Under an argon atmosphere, the terminal bromide carbosilane dendrimer B a 1 1 (0) 4-B r (2 0 · 9 mg '0. 0 4 1 mm ο 1) and tetrasaccharide thioacetamidine body 3 1 (3 0 1 · 2 mg, 0 · 2 2 2 mm ο 1) dissolved in dry DMF 0.3. The alcohol was stirred at 0.3 m 1 '. The previously prepared sodium methoxide (1 M methanol solution) 2 4 4 m 1 ′ was added dropwise and stirred at room temperature for 15.5 hours. Acetic acid 0. 2 m 1 was added, and after neutralizing with sodium methoxylate, the suspension was concentrated. To the reaction mixture were added dried pyridine 4 in 1 and acetic anhydride 2 m 1, and the mixture was stirred at 30 ° C for 3 hours to perform acetylation. The reaction solution was poured into ice water to hydrolyze acetic anhydride, and then extracted with ethyl acetate, and the organic layer was sequentially used with 1 M salt 40 3] 2XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 Acid, Saturated aqueous sodium bicarbonate solution, residual water and brine were washed; strips were dried over anhydrous magnesium sulfate. The organic layer was concentrated, and the residue was purified by GPC (column 2.5 .3 3. 3)) to obtain a B a 1 type 1 carbosilane dendritic structure protected by acetamidine and holding a erythrocyte glycoside ester derivative. Molecules 3 4 (1 2 1.3 mg, 5 5%). Since no molecular peak was observed during molecular weight measurement, a high resolution energy measurement was performed based on the peak value of the divalent ion. &lt; Synthesis of Ball (0) 4-paragloboside-OH (35)>

Under an argon atmosphere, the carrier protected by acetamidine is a B a 1 type 1 carbosilane dendrimer of erythrocyte glycoside ester derivative 3 4 (9 1.3 mg, 0.0 1 7 mm ο 1 ) Dissolved in dry methanol 1.0 m 1, and added sodium ethoxide (4.7 mg, 0.99 mm). After stirring at room temperature for 40 minutes, a 0.1 M sodium hydroxide aqueous solution 5.0 m 1 was added and stirred for 27 hours. After the completion of the reaction, a cationic exchange resin (A Hi Be Er 1 Iter I R 1 2 0 B manufactured by RG A N 0 Company) was added to neutralize the reaction solution. Filter by cotton plug to remove resin and concentrate mash. The residue was purified by gel filtration (G 50, 5% aqueous acetic acid solution) and GPC (GS-2 2 0, 5% aqueous acetic acid solution), and dried to obtain an unprotected compound 3 5 (3 9 1 in g, 68%). Since no molecular peak was observed during molecular weight measurement, a high resolution energy measurement was performed using the peak value of the divalent ion. The identification results are shown below. NMR: 1H (D20, 400MHz): d (ppm) 4.28-4.34 (m), 4.00 (br s), 3. 7 6-3. 8 1 (in), 3. 3 6-3. 7 0 ( in), 3. 1 6 (brs), 2. 4 2 (brs, 1 6 H,-CH '2 S), 1. 8 9 (s, 1 2 Η, NHA c), 1 · 4 9 (brs ), 1 · 3 2 (brs), 0.57 (br s, 8H, S1CH2-). NMR: 13 C (D 2 0, 100 MHZ): d (pp 111) 1 8 1. 2 4, 1 0 2. 6 7, 10 2. 5 8, 41 312XP / Invention Specification (Supplement) / 94-08 / 94] 12338 200538138 10 2 .40, 102.0 1, 8; 1. 84, 7 8.1 7, 7 7.90? 75.0 3, 74.5 7 4. 44, 74.24, 72. 54, 72. 21, 7 1. 89, 70 .67, 69 .95, 69. 64, 68.24, 68. 01, 35. 30, 3 1. 36, 29 .67, 29 .47, 29. 04, 28.58, 24. 67, 23. 80, 22. 04, 11 .38. ESI -MS Ana 1. C 1 3 6 Η 2 &lt; 1 0 N 4 0 8 4 S 4 S i [M + 2 N a] 2 '/ 2: Calculated value 1 7 3 7 · 6 5 4 0' Measured value 1 7 3 7. 6 5 4 4 According to the following reaction formula To prepare Dumbbel 1 (1) 6-paragloboside-0H (37). 31 —〇CH-3 —C2C> DMF / CH3OH ^ \ VcCi〇Ac NHAC OAc c〇Kc 44%% NaOCH3 dlL NaOHaq. ~ Isi \ ^ sw

The yield of CHsOH is 96%. 1/3 Dumbbell (1) 6-parag! 〇bosld ^ 〇H ^ sr

Ke <Synthesis of Dumbbel 1 (1) 6-paragl〇boside-0Ac (36) under an argon atmosphere 'makes the terminal bromide carbosilane dendritic Du in bbe 1 1 (1) 6-B r (2 7 · 9 i】 ig, 〇. 〇3 0 mm ο 1) and tetra-hybrid 3 1 (3 4 5 · 3 n 〖g, 0.2 5 5 mmo 1) dissolved in dry DMF 0.4 丨 n 1 polymer , Tim Li mouth dry methanol 0. 4 m 1, and fully stirred. Drop into a pre-prepared sodium (1 mM methanol solution) 2 80 m 1 and stir at room temperature 1; [小 eye ° f oxygen group

V

The mixture was added with dry pyridine 5 in 1 and acetic anhydride 2.5 5 n 1 and the mixture was neutralized with sodium oxylate at 30 ° C 0. 21Ϊ11. The suspension was concentrated and reacted% \ ί7 &gt; Degree 312XP / Invention Specification (Supplement) / 94-08 / 94112338 42 Therefore, the measurement is performed using the peak value of the trivalent ion. • &lt; Duinbbell (l) 6-paragloboside-0H (37) Synthesis>

200538138 Mix for 4 hours to perform acetamation. The reaction solution was poured into ice water, and after hydrolysis, it was extracted with ethyl acetate. The organic layer was sequentially washed with an acid, a saturated aqueous sodium hydrogen carbonate solution, and a saturated saline solution, and then dried with magnesium sulfate. The organic layer was concentrated, and the residue was purified by 2.5 Η · 3 Η) to obtain D umbbe 1 type 1 carbosilane dendrimer (1 1 0) which was protected by acetamidine and contained a p-glycoside derivative. 0 mg, 44%). At the timing of molecular weight of 1J, due to the absence of argon, no umbbe 1 type 1 carbosilane dendrimer (6 9. E 0. 0 0 8 mm〇1) was dissolved in dry methanol (1.0 m1), and methoxy # 0.07 mm ο 1) was added. After being stirred at room temperature for 30 minutes, a 0.1 M aqueous solution 5.0 m 1 was added and stirred for 5 hours, and then 0.1 M hydroxide 5.0 m 1 was added and stirred for 21 hours. After the reaction was completed, cationic (Amberlite IR120B manufactured by ORGANO) was added to filter the reaction by filtering with a cotton plug to remove the resin, and the filtrate was concentrated. The residual G P C (G S-2 2 0 · G S-3 2 0, 5% aqueous acetic acid solution) was purified to obtain an unprotected compound 3 7 (4 2 · 4 mg, 96%). Since no molecular peak was observed at the time division, it was measured using a high resolution energy based on the trivalent ion. The identification results are shown below. NMR: 1 H (D 2 0, 400 MHZ): d (pp in) 4. 3 3-4. 3 5 (brm), s), 3. 7 9-3. 8 3 (brm) , 3. 5 9-3. 7 0 (brm), 3. 39-3 3. 2 0 (brm), 2. 4 4 (brs, 2 4 H,-CH 2 S), 1. 9 2 (s , 1 312XP / Invention Specification (Supplement) / 94-08 / 94〗 12338 to make 1 M salt of acetic anhydride without GPC (column erythrocytes 36 bubo peak, glucoside esters> mg 5 3 ( 3. 5mg, neutralized by sap exchange with sodium hydroxide aqueous solution. The peak value was determined by freeze-drying the amount of the material. 4.0 2 (br .5 2 (brm), 8 Η, NHA c), 43 200538138 1. 7 7 (brs), 1.5 1 (brs), 1. 35 (br s), 0.49 -〇 · 57 (brm 1 OH 5 S1CH2-.), -0. 14 (1 3 rs, 6H, S 1 CHa ). NMR • 1; 5 C (D 2 0:, 10 OMH z): (d (PP m) 177. 0 8 9, 102. 94, 102.74 10 2. 4 0, 99., 98, 82 .20 , 78. 28, 75 .38, 74. 91, 74 .77, 74. 6 0, 72. ί 39, 72. 78, 72. 56, 72. 25, 7 1.0 2, 70. 41, 70. 26 , 69.! 38, 68. 59, 68. 35, 61 · 07, 60.8 5, 59. 95, 5 5. 28, 29.! 54, 29. 05, 25. 13, 24. 26, 22.4 3, 20. 68, -0. 25. ’

ESI-MS Anal.C2l2H378N6〇12GS6Si3 [M + 3Na] 3 + / 3: Calculated value 1 7 9 0.6 9 1 6 and found value 1 7 9 0. 6 8 9 1 · (Example 4) &lt; 1.1 Study on the appropriate concentration of dengue virus> After culturing K 5 62 cells to full proliferation (c ο nf 1 uent), they were collected in microtubes at 4 X 105 cells / tube. This was centrifuged at 3,600 r p in at 4 ° C for 3 minutes, and the obtained cell pellets (pe 1 1 e t) were washed once with P B S 1 m 1 containing 0.1% B S A. After centrifugation at 3 ° C, 3,600 rpm for 3 minutes, the dengue virus (T h N Η-7/9 3) stock solution was diluted to 20,000 with PBS containing 0.1% BSA. Units / m 1, 1 0 0 0 units / m 1, 5 0 0 units / m 1, 2 50 0 units / m 1, and 1 2 5 units / m 1, and adding 1 0 0 // 1 K 5 6 2 cells, react at 4 ° C for 1 hour. Press again, it is virus (-), only add the same amount of P B S containing 0.1% B S A. After the reaction was completed, the cells were washed three times with cold PBS (-) 1 m 1, and A 1 e X a labeled mouse anti-dengue virus monoclonal antibody (c 1 ο ne 1 2 D 1) was added as a primary antibody, respectively. 1/7 E 8) diluted to 3. 1 3 // g 44 312XP / Invention Specification (Supplement) / 9108/94]] 2338 200538138 / m 1 concentration of antibody solution 1 0 0 // 1 to make it full After suspension, the reaction was carried out at 4 ° C for 30 minutes. As a control stain, A 1 e X a

Mark the rat IgG1 to the same concentration. After the reaction was completed, the cells were washed three times with cold PBS (-) 1 ml, and 0.5 m 1 of cold PBS (-) was added to the obtained pellets, and the cells were fully suspended, and flow cytometry was performed. The measurement was performed with a f10wcy meter (Epics, USA). The results of measuring the binding of dengue virus to the cell surface used in the study of the appropriate concentration of dengue virus are shown in Fig. 1. It is known from FIG. 1 that the virus is between 125 and 2000 units / m 1, which fully shows the concentration-dependent binding property of the virus to K 562 cells. Therefore, regarding the concentration of the virus used for the measurement, the concentration of the virus used for the subsequent reaction was determined to be 1,000 units / m 1 in consideration of both the sensitivity of detection sensitivity and virus dependence. In subsequent experiments, the virus concentration was set to 10 ◦ 0 units / m1. &lt; 1.2 Study on the appropriate concentration of Alexa labeled primary antibody (12D11 / 7E8)> Based on the results of 1-1, the dengue virus concentration was set to 1000 units / ml to determine the binding property to the cell surface. Here, the concentration of the primary antibody was adjusted to 1.5 5 6-1 3 · 5 // g / m 1 with cold P B S (-). As a control staining, A 1 e X a labeled mouse I g G 1 was prepared to the same concentration. The experimental method is compared with 1-1. In future experiments, the primary antibody concentration was set to 6. 25 μg / m. The dengue virus pair used in the study of the appropriate concentration of A 1 e X a labeled primary antibody (12011/7 £ 8) was used. The measurement results of cell surface binding are shown in FIG. 2. According to the results of FIG. 2 and the research results of the antibody concentration used in the measurement, it is known that the binding amount is between the primary antibody concentration of 1.5 6-6. 2 5 // g / m 1 and depends on the antibody concentration. And 45 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 changed. However, when the concentration of the antibody is higher, for example, no significant change in binding is observed at 13.5 / g / m1, so it is judged that the detection of all virus particles bound to the cell surface, 6. 25 The concentration of the detected antibody in g / η〗 1 is a sufficient and sufficient concentration. Therefore, it was decided that the appropriate antibody concentration for the measurement was 6.25 // g / ml. `` Study on Inhibitory Activity of Dengue Virus on 丨 (562 Cell Surface Binding Inhibition by 1-3 Dumbbell (l) 6-paragl〇b〇side-0H〉

The effects of the addition of Diuibbell (l) 6-paragloboside-0H produced in Example 3 on the binding of dengue virus to the surface of K 5 62 cells were studied using the appropriate virus and appropriate antibody concentrations determined in 1-1 and 1-2. The study was performed with a virus concentration of 1,000 units / ml and an antibody concentration of 6.25 // g / ml. First, the Dumbbe 1 1 dendrimer and n L c 4 (G a 1 1-4 G 1 c NA c / 3 1-3 G a 1/3 1-4 G 1 c) oligosaccharide PBS containing 0.1% BSA was prepared as 1 2 5-1 0 ◦ 0 # M concentration solution and 50 &quot; 1 per micro tube. A dengue virus solution 50 &quot; 1 was added to the microtube, and after thorough mixing, preculture was performed at 4 ° C for 30 minutes. The preculture solution was added to a microtube containing K 5 62 cells (2 X 105 cells / tube), and after thorough mixing, virus binding was performed at 4 ° C for 1 hour. Only the same amount of P B S containing 0.1% B S A was added to the virus (-) sample. Subsequent experimental operations are compared with 1-1. Dumbbe 1 1 (1) 6-parag 1 〇b 〇side-〇 (Dengue virus pair at the time of addition) (5 6 2 The measurement results of cell surface binding properties are shown in FIG. 3. As described above, in Duinbbell (l) 6-paragloboside-〇H, the binding activity of dengue virus on the surface of K 5 62 cells was measured to study the effect of Dumbbell (l) 6-paragloboside-0H. As shown in Figure 3, 46 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138

In the presence of 〇111111) 1:) 11 (1) 6-1;) 8 1'8 and 101031 (16-01'1), a concentration-dependent inhibition of dengue virus binding to the cell surface was observed The effect was observed at 66% inhibition at the highest concentration of 1000 V Μ used in the experiment, and at 50% inhibition at the lowest concentration of 12 5 // Μ. TLC binds to a glycolipid with nLc 4 sugar chain called erythrocyte glycoside ester, and this glycolipid inhibits virus binding to the surface of K 5 62 cells. The n L c 4 used this time is only The oligosaccharide molecule of the sugar chain portion of the erythrocyte glycoside ester. When the same concentration (50 0 # Μ) as that of the Dumbbell (1) 6-paragloboside-OH used for the inhibition, it was confirmed that only the oligomerization of the sugar chain portion The sugar has almost no inhibitory effect, and shows that the sugar chain is formed into a polyvalent addition of Du in bbe 1 1 type molecular structure, and the fact that the inhibitory effect is exhibited and strengthened σ (Example 5) (Dengue virus infection Inhibition test) The β κ 2 1 cells prepared at 2.3 X 105 cells / m 1 were dispersed in a 4 8 we 1 1 plate to 250 // 1 / well, and incubate for 20 to 24 hours. Second, after removing the culture medium, the dengue virus solution (5 2 0 0 FFU / in 1) 2 0 0 # 1 and the inhibitor solution shown in Table 1 2 0 The mixed solution obtained by mixing 0 # 1 was inserted into the 4 8 we 1 1 p 1 ate at a force of 1 2 5 μ 1 / we 1 1. Then, the infection was performed at 37 ° C. for 2 hours. After the infection was completed, SF-D Μ E Μ 3 0 0 β 1 / we 1 1 Wash the strips 3 times, add 0ver 1 aymedium, and incubate for 4 3 hours. After the end of the culture, fix with trimeraldehyde, and then perform NP-4 0 treatment. Secondly, an antibody (dengue haemorrhagic fever patient serum) response was performed once, followed by a secondary antibody (HRP-G a Η I g 'G) response. Second, κ nica immunized 47 3] 2XP / Description of the Invention (Supplement) / 94-08 / 94112338 200538138 Staining k 1 ΐ Staining ◦ The dengue virus infection inhibition test results obtained as above are shown in Tables 1 and 4. In addition, in Table 1, the infection rate ( %) Is the Focus number of each test relative to F ocusnumber Α ν erage of test A Percentage of Average. Table 1 Test virus inhibitor solution Focus number Average Soil SD infection rate inhibitor concentration (%) A + None-58. 67 9. 71 100 B + Ball (0) 4 (35) &quot; ImM 44. 33 4. 93 75.55 C + Ball (0) 4 (35) &quot; 0.5ηι 50. 67 7.37 86.36 D + Fan (0) 3 (33) 2) ImM 31.33 5. 69 53.40 E + Fan (0) 3 (33) 2) 0.5 mM 39. 67 5. 13 67.61 F + Dumbbell (l) 6 (37) 3; ImM 27. 67 4. 93 47. 16 ~ G + Dumbbell (l) 6 (37) 3) 0.5 mM 31. 67 3. 21 53.97 Η + Lacto-N-neotetraose ImM 61. 33 2. 08 104. 5 I + Lacto-N-neotetraose 0.5 mM 57. 33 6. 66 97.71 J + Heparin 100 u R / ml 1.33 1. 53 2.26 Κ-None-0-0 Ball (Q) 4-paragloboside-〇H (35) obtained in Example 3

) Fan (〇) 3-paragloboside-OH (33) obtained in Example 3) Duuibbell (l) 6-paragloboside-〇H (37) obtained in Example 3 [Schematic description] Figure 1 shows the dengue virus A graph of the results of measuring the binding of dengue virus to cell surfaces used in studies at appropriate concentrations. Figure 2 is a graph showing the results of measuring the binding of dengue virus to the cell surface used in the study of the appropriate concentration of Alexa-labeled primary antibody (12D11 / 7E8). Figure 3 is a graph showing the results of measuring the binding of dengue virus to the surface of K 5 62 cells when Dumbbell (l) 6-paragloboside-0H was added. Fig. 4 is a graph showing the experimental results of dengue virus infection inhibition. 48 312XP / Invention Specification (Supplement) / 94-08 / 94112338

Claims (1)

200538138 10. Scope of patent application: 1. A carbosilane dendrimer carrying sugar chains, which is represented by the following general formula (1); (R 1), „S 1 {-R 2-S i ( R 6) 1 [R 3-S i (R 7) k (R 4-S-R 5-A) 3-k] 3 -1} n (1) (where 1 ^, 1 ^ and R7 are Any of alkyl, phenyl, vinyl, and allyl groups having 1 to 6 carbon atoms may be the same or different. R2, R3, R4, and R5 are alkyl groups having 1 to 6 carbon atoms. Any of phenylene and alkenyl, which may be the same or different, A is the following chemical formula Parag 1 〇b 〇side derivatized group, alpha mannosyl group, or terminal 1 position of oligosaccharide formed from mannose containing only 2 to 8 mannose chains as shown The hydroxyl group is a group except for a hydrogen atom, m is an integer of 0 to 3, η is an integer of 1 to 4, m + n is 4, and k and 1 are any of 0 to 2, k and 1 may be Same or different). 2. A kind of carbon-stone-containing dendritic and molecular structure containing sugar bonds, which is represented by the following general formula (2); (R 1), n S i [-R 2-S i (R 6) 1 (R4-S-R 5-A) 3 -1] n (2) (wherein the definition of ruler 1, ruler 2, ruler &lt; 1, ruler, ruler (), eight, 11], 11 and 1 As previously mentioned). 3, —Carbon stone with sugar chains :): complete dendrimer, which is represented by the following general formula (3); (R 1), „S i (R 4-S-R 5- A) η (3) (wherein R1, R'1, R5, A, m and η have the same definitions as above) 49 312XP / Invention Specification (Supplement) / 94-08 / 94112338 200538138 4. If applying for a patent Carbosilane dendrimers with sugar chains in any one of the scope of items 1 to 3, where A is the pair of erythrocyte glycoside esters' derivative groups. The carbosilane-containing dendrimer having a sugar chain according to any one of 3 items, wherein A is the α-mannose piperanyl group, or from mannose containing only 2 to 8 mannose chains The hydroxyl group at the terminal 1-position of the formed oligosaccharide removes the hydrogen atom group. 6. The φ-carbosilane silyl dendrimer having a sugar chain as described in any one of claims 1 to 3, wherein, In the formula, A is the alpha mannosyl group or a group represented by the following chemical formula (2) 50 200538138 A method for producing a dendritic polymer, characterized in that the following general formula (6) (R1) S 1 {-R 2-S 1 (R l)) 1 (R 4-X) 3-.} η (6) &quot; (wherein, R1, R2, R4, Rϋ, πl, n and l have the same definitions as above, X represents a • I element atom) and a halogen compound represented by the general formula (5) The sulfur compounds react. 9. A method for manufacturing a carbosilane-containing dendrimer having a sugar chain as described in item 3 of the scope of the patent application, characterized in that the following general formula (7) (R 1), S i (R 4 -X) η (7) (wherein R1, R4, m, and η have the same definitions as above, and X represents a halogen atom) The halogen compound represented by the general formula (5) reacts with the sulfur compound represented by the general formula (5). 1 0 A kind of dengue virus infection inhibitor, which is characterized in that it contains a carbosilane dendrimer with a sugar chain as an active ingredient as described in item 4 of the patent application scope. 1 1. An antiviral agent, Contains as an active ingredient the carbon stone yaki dendrimer with sugar bonds as the active ingredient as in item 4 of the scope of the patent application. ^ 1 2. —A substance for screening anti-rhenium IV agents, characterized in that it contains Carbosilane dendrimers with sugar chains as the active ingredient in the scope of the patent application No. 5 51 312XP / Invention Specification (Supplement) / 94-08 / 9411233 8