JP2007515434A - Glycosaminoglycan (GAG) mimics - Google Patents

Abstract

  The present invention relates to compounds designed to mimic the structure of GAG, methods for preparing these compounds, compositions containing these compounds, and anti-angiogenesis, anti-tumor metastasis, anti-inflammatory, anticoagulant activity in mammals , To the use of these compounds for antithrombotic and / or antimicrobial treatment and compositions comprising these compounds.

Description

  The subject of the present invention belongs to the technical field of compounds that mimic the structure of specific carbohydrates. Specifically, the present invention belongs to the technical field of glycosaminoglycan (GAG) mimics.

  In particular, the present invention relates to compounds comprising at least one reactive functional group designed to mimic the structure of GAG. The present invention also provides methods for preparing these compounds and compositions containing these compounds, as well as for anti-angiogenesis, anti-tumor metastasis, anti-inflammatory, anticoagulant action and / or antimicrobial treatment in mammals. It relates to the use of these compounds and compositions containing them. The invention also relates to the use of these compounds and compositions comprising them in the treatment of mammals susceptible to these factors.

  Glycosaminoglycans (GAGs) are linear polyanionic polysaccharides that are produced in most animal cells and are often attached to protein nuclei [1, 2]. GAGs are abundant in production (like proteoglycans) and are pushed to the cell surface by cells and migrate to the extracellular matrix (ECM) [3]. Among GAGs, in particular, GAG belonging to the heparan sulfate (HS) family (HS-GAG) mediates a myriad of physiological processes. For example, HS-GAG plays an important role in cell growth and development, angiogenesis, coagulation, tumor metastasis, cell adhesion, growth factor activation, cytokine and chemokine binding, and infection by bacteria and viruses [4-6]. In recent years, the number of entries in the list of proteins that interact with GAG has increased dramatically, and the protein is still added to this list. It has recently become clear that sequences unique to extracellular GAGs bind specifically to important proteins, thereby affecting basic biological processes.

  It can bind to GAG-binding protein, and its biological activity, for example, activation of AT-III by various pentasaccharides [7, 8], and fibroblast growth factor by polysulfated sugar chain Molecules that can regulate activities such as activation [9], mimic certain GAG structures and are also referred to as “GAG mimetics” are known. Similarly, it is also known that GAG mimics can dissociate the binding between GAG and its target protein, and thereby inhibit the biological or disease function of the protein. . For example, anti-cancer agents developed in connection with targeted HS-binding angiogenesis factors include polysulfonated compounds [10], suramin and related slagisters [11], and sulfates [12, 13]. Yes.

  The present invention relates to a novel small molecule GAG mimic that binds to and regulates the action of GAG binding proteins. These compounds contain at least one negatively charged functional group (preferably a sulfo group) to interact with a positively charged residue at the GAG binding site of the target protein, It also includes one or more substituents that facilitate interaction with other protein residues in and near the aforementioned binding site. Important and unique features of the compounds described herein are that they have few sulfo groups and have a lower molecular weight than the polysulfated GAG mimics described above, such as sulfated oligosaccharides [ 12, 13]. Another important feature is that the chemical structure of a cyclic skeleton (eg, monosaccharide) having a sulfo group and other substituents located according to a predetermined characteristic orientation around the ring structure. Based on this, the structure is inconsistent with the disorderly charged simple GAG mimic [14] reported by Xylevsky. Binding of the compounds described herein to HS-binding angiogenic factors is verified using a surface plasmon resonance (SPR) solution affinity assay. In addition, selection of compounds for inhibiting HS-mediated cell infection and herpes simplex virus cell-to-cell propagation has been performed.

  One aspect of the present invention relates to the use of the eel reaction [15, 16] resulting in various configurations of GAG mimics. In addition to various aspects in which each charged chemical structure is bonded to each other or to other functional groups, the gist of the present invention that mimics various structures of GAG is also described. As is obvious to those skilled in the art, the means for imparting functionality to the cyclic skeleton is not limited to the eel reaction. For example, many reaction systems including alkylation, acylation, and cycloaddition are used.

  An object of the present invention is to provide a charged compound, that is, a novel compound having utility as a GAG mimic.

  It is a further object of the present invention to provide an efficient synthetic route for these compounds.

  According to a first aspect of the present invention, the following formula:

式中、ｎは、０から２の整数であり、
Ｚは、Ｎ、Ｎ(Ｏ)、Ｏ、Ｓ、Ｓ(Ｏ)、Ｓ(Ｏ)₂、Ｐ、Ｐ(Ｏ)、Ｐ(Ｏ)₂、Ｓｉ、Ｓｉ(Ｏ)またはＳｉ(Ｏ)₂であり、
独立的な各Ｘは、Ｃ、Ｃ(Ｏ)、Ｎ、Ｎ(Ｏ)、Ｏ、Ｓ、Ｓ(Ｏ)、Ｓ(Ｏ)₂、Ｐ、Ｐ(Ｏ)、Ｐ(Ｏ)₂、Ｓｉ、Ｓｉ(Ｏ)またはＳｉ(Ｏ)₂、または化学結合であり、および
独立的なＲ₁〜Ｒ₆の各々は、化学結合、あるいは、水素；ハロゲン；直鎖、環状、分枝、置換、ヘテロ環状、ヘテロ原子置換または未置換のアルキル、アルケニル、アルキニル、アリールまたはヘテロアリール；-Ｏ-Ｐ-(Ｓ)(ＯＨ)₂で表されるリン酸塩、チオリン酸塩のようなホスホリル基；-Ｏ-Ｐ-(Ｏ)(ＯＲ)₂で表されるリン酸エステル；-Ｏ-Ｐ-(Ｓ)(ＯＲ)₂で表されるチオリン酸エステル；-Ｏ-Ｐ-(Ｏ)ＯＨＲで表されるホスホン酸塩；-Ｏ-Ｐ-(Ｓ)ＯＨＲで表されるチオホスホン酸塩；-Ｏ-Ｐ-(Ｏ)ＯＲ₁Ｒ₂で表される置換ホスホン酸塩；-Ｏ-Ｐ-(Ｓ)ＯＲ₁Ｒ₂で表される置換チオホスホン酸塩；-Ｏ-Ｐ-(Ｓ)(ＯＨ)(ＳＨ)；および、環状リン酸塩；-Ｏ-Ｐ-(ＯＲ)-ＮＲ₁Ｒ₂で表されるフォスフォアミダイト；-Ｏ-Ｐ-(Ｏ)(ＯＲ)-ＮＲ₁Ｒ₂で表されるフォスフォアミダイトなどのリン含有化合物；-Ｏ-Ｓ-(Ｏ)(ＯＨ)、-ＳＨ、-ＳＲ、-Ｓ(→Ｏ)-Ｒ、-Ｓ(Ｏ)₂Ｒ、ＲＯ-Ｓ(Ｏ)₂ ^-、-Ｏ-ＳＯ₂ＮＨ₂、-Ｏ-ＳＯ₂Ｒ₁Ｒ₂、または-ＮＨＳＯ₂ＮＨ₂で表されるスルファミドのようなイオウ含有基；-ＮＨＲ、-ＮＲ₁Ｒ₂、-ＮＨＡｃ、-ＮＨＣＯＲ、-ＮＨ-Ｏ-ＣＯＲ、-ＮＨＳＯ₃、-ＮＨＳＯ₂Ｒ、-Ｎ(ＳＯ₂Ｒ)₂などのアミノ基、および／または、-ＮＨ-Ｃ(=ＮＨ)ＮＨ₂などのアミドノ基、および／または、-ＮＨ-ＣＯ-ＮＲ₁Ｒ₂などのウレイド基、または-Ｈ-Ｃ(Ｓ)-ＮＨ₂などのチオウレイド基；化学式Ｉで表されるその他の単位構造、すなわち、式中、前述したＺ、ＸおよびＲ₁〜Ｒ₆が、任意の位置で結合してなる単位構造；または、以下の式、すなわち；

Where n is an integer from 0 to 2,
Z is N, N (O), O, S, S (O), S (O) ₂ , P, P (O), P (O) ₂ , Si, Si (O) or Si (O) ₂ And
Each independent X is C, C (O), N, N (O), O, S, S (O), S (O) ₂ , P, P (O), P (O) ₂ , Si , Si (O) or Si (O) ₂ , or a chemical bond, and each of the independent R _{1 to} R ₆ is a chemical bond or hydrogen; halogen; linear, cyclic, branched, substituted, Heterocyclic, heteroatom-substituted or unsubstituted alkyl, alkenyl, alkynyl, aryl or heteroaryl; phosphoryl groups such as phosphates and thiophosphates represented by —O—P— (S) (OH) ₂ ; -O-P- (O) (OR) ₂ phosphate ester; -O-P- (S) (OR) ₂ thiophosphate ester; -O-P- (O) OHR A phosphonate represented by —OP— (S) OHR; a substituted phosphonate represented by —O—P— (O) OR ₁ R ₂ ; (S) Substituted thiophosphones represented by OR ₁ R ₂ -O-P- (S) (OH) (SH); and cyclic phosphates: phosphoramidites represented by -OP- (OR) -NR ₁ R ₂ ; -OP Phosphorus-containing compounds such as phosphoramidite represented by — (O) (OR) —NR ₁ R ₂ ; —O—S— (O) (OH), —SH, —SR, —S (→ O) — Like sulfamide represented by R, —S (O) ₂ R, RO—S (O) ₂ ⁻ , —O—SO ₂ NH ₂ , —O—SO ₂ R ₁ R ₂ , or —NHSO ₂ NH ₂ Sulfur-containing groups; amino groups such as —NHR, —NR ₁ R ₂ , —NHAc, —NHCOR, —NH—O—COR, —NHSO ₃ , —NHSO ₂ R, —N (SO ₂ R) ₂ , and An amido group such as —NH—C (═NH) NH ₂ ; and / or a ureido group such as —NH—CO—NR ₁ R ₂ , or a thioureido such as —H—C (S) —NH _2. A group; other unit structures of formula I, ie, the formula Wherein Z, X and R _{1 to} R ₆ described above are bonded at any position; or the following formula:

で表現される官能基に基づいた化学構造であって、式中、Ｙは、化学結合、または、直鎖、環状、分枝、置換、ヘテロ環状、ヘテロ原子置換または未置換のアルキル；直鎖、環状、分枝、置換、ヘテロ環状、ヘテロ原子置換または未置換のアシル；および、アリール、置換アリール、ヘテロアリール；および、独立的なＲ₇〜Ｒ₁₁の各々は、少なくとも一つの化学式Ｉに記載の化学構造、または、化学式IIに記載の化学構造、すなわち、Ｚが酸素で、かつＸが酸素または化学結合である場合に、Ｒ₁〜Ｒ₅のすべてが、水素またはＣＨ₂ＯＨでなく、また、Ｚが窒素で、かつＸが酸素または化学結合である場合に、Ｒ₁〜Ｒ₆のすべてが、水素でないとの条件を満足する化学式IIに記載の化学構造からなるグループから選択される式で表される化合物が提供される。

Wherein Y is a chemical bond or a linear, cyclic, branched, substituted, heterocyclic, heteroatom-substituted or unsubstituted alkyl; linear , Cyclic, branched, substituted, heterocyclic, heteroatom-substituted or unsubstituted acyl; and aryl, substituted aryl, heteroaryl; and each of the independent R ₇ -R ₁₁ is at least one of Formula I Or the chemical structure described in Formula II, ie, when Z is oxygen and X is oxygen or a chemical bond, all of R _{1 to} R ₅ are not hydrogen or CH ₂ OH. And when Z is nitrogen and X is oxygen or a chemical bond, all of R _{1 to} R ₆ are selected from the group consisting of the chemical structures described in Formula II that satisfy the condition that they are not hydrogen. Represented by Compounds are provided.

  According to a second aspect of the present invention, a pharmaceutical or veterinary medicine for preventing or treating a disease caused by angiogenesis, tumor metastasis, inflammation, coagulation, thrombosis and / or microbial infection in a mammal Compositions are provided, ie comprising a pharmaceutically or veterinarily acceptable carrier or diluent for at least one said compound together with at least one compound according to the first aspect.

  According to a third aspect of the present invention, there is provided a medicament for the prevention or treatment of diseases caused by angiogenesis, tumor metastasis, inflammation, coagulation, thrombosis and / or microbial infection in mammals. Uses of the compounds described in the first aspect are provided.

  According to a fourth aspect of the present invention, there is provided a method for preventing or treating a disease caused by angiogenesis, tumor metastasis, inflammation, coagulation, thrombosis and / or microbial infection in a mammal, There is provided a method comprising administering to the mammal an effective amount of at least one compound according to one embodiment or a composition comprising at least one such compound.

  According to another aspect of the present invention, there is provided a process for synthesizing the compound described in the first aspect.

Furthermore, with respect to the compounds described in the first embodiment, unless otherwise specified, alkyl groups, aryl groups, and other substituents are used in accordance with normal recognition in the art. For example, an alkyl group or an aryl group usually has 1 to 10 carbon atoms. In addition, two functional groups of R _{1 to} R ₅ can be bonded to each other to form a bicyclic chemical structure, or the cyclic structure represented by Formula I has a double bond That is, two adjacent functional groups in XR _{1 to} XR ₅ can also be bonded.

  Preferred compounds of the present invention have the structural formulas shown in Tables 1 to 4 below.

  In order to facilitate an understanding of the present invention, and for practical application, one or more preferred embodiments are disclosed for illustrative purposes.

  In the present specification, the following abbreviations are used.

GAG Glycosaminoglycan HS Heparan sulfate FGF Fibroblast growth factor aFGF Acidic fibroblast growth factor (or FGF-1)
bFGF basic fibroblast growth factor (or FGF-2)
VEGF Vascular Endothelial Growth Factor SPR Surface Plasmon Resonance HSV Herpes Simplex Virus The present inventors have found that a number of compounds having GAG mimetic properties can be synthesized by various procedures shown in the following examples. It has come. These compounds can be effectively used in the prevention or treatment of diseases caused by angiogenesis, tumor metastasis, inflammation, microbial infection, coagulation or thrombosis in mammals. The utility resulting from the action of these compounds extends to the modulation of the activity of GAG binding proteins corresponding to the disease process.

  As described above, the GAG mimetic of the present invention can be synthesized with various routes including the eel reaction, and the synthesis process usually includes a sulfonation step.

  Suitable compounds in the first aspect of the present invention described above include compounds defined by Structural Formulas I and II and compounds shown in Tables 1 to 4 below.

  As described above, the compound of the present invention can be effectively used in the prevention or treatment of diseases caused by angiogenesis, tumor metastasis, inflammation, microbial infection, coagulation or thrombosis in mammals. These compounds can be effectively used particularly in the treatment of the aforementioned diseases in humans. Generally, these compounds are administered as a component of a pharmaceutical composition detailed below.

  Pharmaceutical compositions for oral administration can be in tablet, capsule, powder or liquid form. Tablets can contain a solid carrier such as gelatin, and adjuvants or inert diluents. Liquid pharmaceutical compositions can usually contain a liquid carrier such as water, petroleum, animal or vegetable oils, vegetable oils, mineral oils or synthetic oils. In addition to physiological saline, glycols such as ethylene glycol, propylene glycol or polyethylene glycol can also be contained. These compositions and preparations usually contain at least about 0.1% by weight of the compounds of the invention.

  Parenteral administration includes the following routes: intravenous, cutaneous or nasal, intramuscular, intraocular, epithelial, intraperitoneal, and topical. As topical administration, in addition to administration to the skin, eyes, rectum and nose, there are administrations using inhalation means and aerosol means. For parenteral, skin or subcutaneous injection or injection into the site in need of treatment, the active ingredient is pyrogen-free and has a suitable pH, isotonicity and stability, parenterally Can be in the form of an aqueous solution that is acceptable. A person skilled in the art will be able to prepare a suitable solution using, for example, the compound of the present invention or a derivative thereof.

  In addition to at least one compound and a carrier or diluent, the composition of the present invention may contain pharmaceutically or veterinarily acceptable excipients, buffers, stabilizers, tonicity agents, preservatives or antioxidants. Agents or other materials well known to those skilled in the art can further be included. One skilled in the art will readily appreciate that these materials are non-toxic and do not interact with the effects that the compounds exert. The properties required of the additive vary depending on the route of administration of the composition, i.e., whether the composition is administered orally or parenterally. In order to maintain the pH of the aqueous composition within the approximate range of physiological pH, or in the range of about 5.0 to about 8.0, the aqueous composition usually contains a buffer.

  In addition to at least one compound, the composition of the present invention may further contain an active ingredient. These components are selected primarily for their anti-angiogenesis, anti-tumor metastasis, anti-inflammatory, anti-coagulant action, and antimicrobial effects, but can also be selected based on their effect on the associated condition.

  The pharmaceutical or veterinary composition of the present invention is administered to a patient in a prophylactically effective amount or a therapeutically effective amount required in view of the individual disease state. The actual dose of at least one compound administered in the form incorporated into the composition will depend on the nature and severity of the disease for which the therapeutic or prophylactic treatment is being performed. Preparation of a prescription including determination of dose etc. is a matter to be performed by the doctor or veterinarian treating the patient. However, when the composition is administered to a human, usually from about 0.01 to about 100 mg / kg body weight, preferably from about 0.1 to about 10 mg / kg body weight of the compound is administered.

These compounds can be incorporated into the compositions as pharmaceutically or veterinary acceptable derivatives thereof. “Derivatives” of the compounds described herein include salts, coordination complexes having metal ions such as manganese ions (Mn ²⁺ ) and zinc ions (Zn ²⁺ ), in vivo hydrolysable esters Esters, free acids or bases, hydrates or prodrugs. Compounds with acidic groups such as phosphates and sulfates form salts with alkali metals and alkaline earth metals such as sodium, potassium, magnesium and calcium, and triethylamine and tris (2-hydroxyethyl) amine be able to. Compounds having basic groups such as amines can also form salts with inorganic acids such as hydrochloric acid, phosphoric acid or sulfuric acid, or organic acids such as acetic acid, citric acid, benzoic acid, fumaric acid or tartaric acid. it can. A compound having both an acidic group and a basic group can include salts.

  Esters can be formed between hydroxyl groups or carboxylic acid groups within the compound and suitable carboxylic acid or alcohol reactive elements using techniques well known in the art.

The prodrug derivative of the present invention can be modified into the parent compound in vivo or in vitro . At least one biological activity of the parent compound is usually inhibited when in the prodrug form, and can also be activated by converting the prodrug to the parent compound or a metabolite thereof. Examples of these prodrugs are glycolipid derivatives having one or more lipid moieties that are cleaved with an enzyme having phospholipase activity to impart a substituent that induces the release of the free form of the compound. In prodrugs of the compounds of the invention, a protecting group may be used that is either removed in vivo to release the active compound or plays a role in inhibiting the disappearance of the drug. Suitable protecting groups are well known to those skilled in the art, and acetate groups and the like can be used.

  As described above, the compound of the present invention is effective in the manufacture of a medicament for preventing or treating diseases caused by angiogenesis, tumor metastasis, inflammation, coagulation, thrombosis and / or microbial infection in mammals. Available. Such pharmaceutical production processes are well known to those skilled in the art, and the processes used to produce the pharmaceutical compositions described above can be used.

  Compounds belonging to the present invention have been found to bind to growth factors. In particular, it has been confirmed that these compounds show affinity for aFGF, bFGF and VEGF. Thus, these compounds can be used as anti-angiogenic agents, anti-tumor metastasis agents and / or anti-inflammatory agents in the treatment of mammals including humans. Applications of these compounds include the treatment of angiogenesis-dependent diseases such as angiogenesis involving the growth of solid tumors and proliferative retinopathy, and the treatment of inflammatory diseases and conditions such as rheumatoid arthritis. These compounds also activate growth factors and can be used in treating cardiovascular.

  As described above, the compound of the present invention can also be used for anticoagulants or antithrombotic agents. Therefore, these compounds have been used in many thrombosis and cardiovascular diseases, including the most well-known diseases such as deep vein thrombosis, pulmonary embolism, thrombotic stroke, peripheral arterial thrombosis, unstable angina and myocardial infarction. It can be used in both prevention and treatment. Since compositions containing charged amino acid compounds can be delivered orally, it is practical to use these compounds as an alternative to warfarin, an anticoagulant for oral administration that exhibits severe side effects. You can say that.

  Furthermore, since it has been found that the compounds of the present invention inhibit viral infection, these compounds can also be used as antiviral agents used for the prevention or treatment of many viral infections.

  The compounds of the present invention treat or prevent infectious diseases caused by pathogens that use HS as an adhesion / invasion receptor [6], such as HSV, HIV, dengue virus, yellow fever virus, cytomegalovirus and hepatitis C virus. Can be used particularly preferably. Similarly, the compounds of the present invention can also be suitably used in the treatment or prevention of infectious diseases caused by non-viral microbial pathogens that utilize HS as an adhesion / invasion receptor, such as plasmodium (malaria). It is noteworthy that the compounds of the present invention inhibit HSV-1 and HSV-2 propagation between cells.

  Following the outline of the present invention, the compounds of the present invention, their synthesis, and their biological activities will be exemplified below with reference to the tables attached to the present specification. It is not limited.

General procedure
General Procedure for Alkylation and Deprotection of Diols DMF containing diol (1 eq) is added dropwise to a cooled stirred suspension of DMF containing prewashed (hexane) sodium hydride (5 eq). It was. Once the addition was stopped, stirring was continued (0 ° C. → room temperature, 20 minutes). The resulting mixture was cooled (0 ° C., 5 minutes) and alkyl halide (2 eq) was added dropwise (0 ° C. → room temperature, overnight) with continued stirring. After the mixture was cooled again, methanol (5 ml) was added with continued stirring (5 minutes). The solvent was evaporated and the residue was subjected to analysis (ethyl acetate) and flash chromatography and then checked for homogeneity (TLC). The residue was co-concentrated (2 x 10 ml methyl cyanide). Methanol / methyl cyanide (1: 1) containing the crude mixture (50 mg) and p-TsOH.H ₂ O (50 mg) was heated under reflux (1 hour). The resulting mixture was cooled (room temperature) and triethylamine (100 μl) was added before the solvent was evaporated. The residue was applied to flash chromatography (ethyl acetate / hexane) to give the diol.

General Procedure for Sulfonation of Alcohols DMF containing a mixture of alcohol and SO ₃ trimethylamine (2 eq / hydroxyl) was heated (60 ° C. overnight). The cooled (room temperature) reaction mixture was treated with methanol and then sodium carbonate (10% w / w) was added to adjust to basic (pH> 10). The mixture was filtered and the filtrate was concentrated and co-concentrated (with water). When deacetylation of the sulfate was required, the crude product was dissolved in water and 1M sodium hydroxide (2 eq / acyl group) was added. Upon completion of removal of the protective element, the product was transferred to the next stage of processing. Water containing crude sulfate was applied to size exclusion chromatography. The pure fractions were concentrated and co-concentrated (with water) and lyophilized (with water) to obtain the sulfate. If necessary, after lyophilization, the product is passed through an ion exchange resin column (AG ⁽ registered trademark ⁾ -50W-X8, sodium ion type, 1 x 4 cm, deionized water, 15 ml) to homogenize the product. In the form of sodium salt. The collected solution was concentrated and lyophilized to obtain the final product as a colorless glass or white foam.

Size exclusion chromatography
Size exclusion chromatography (SEC) was performed using Bio-Gel P-2 equipped with a 5 × 100 cm column with 0.1M NH ₄ HCO ₃ flowing at a flow rate of 2.8 ml / min and 2.8 minutes (7.8 ml ) Fractions were collected. These fractions are analyzed for carbohydrates by TLC (carbonization) and / or charged molecules by dimethylmethylene blue test, and in these, by salt purity analysis by capillary electrophoresis (CE). What was thought to be not was collected and lyophilized.

  LH20 SEC process (2 x 95 cm, deionized water, 1.2 ml / min, per vial) in the presence of incomplete sulfation by-products or salt impurities (usually present in small quantities but frequently detected) 3.5 minutes) was applied to remove them completely.

Dimethylmethylene blue test A dimethylmethylene blue (DMB) reagent was prepared by dissolving 16 mg DMB in 1 liter deionized water containing 3.04 g glycine and 2.37 g sodium chloride. 0.1M hydrochloric acid (95 ml) was added to adjust the pH to 3.0. Stock solutions were stored in brown bottles at room temperature (this solution was stable for at least 3 months under these conditions). Ten μl of the fraction solution was placed in each well of a 96-well microtiter plate. 55 μl of DMB stock solution was added to each well where the sample was placed. The immediate color change from blue to pink indicated the presence of charged molecules, ie the sulfated product fraction.

General Procedure for NIS Glycosylation Glycosyl acceptor (1 eq), thioglycoside donor (1.1 eq), 500 mg of freshly activated powdered trigonal sieve and 10 ml of dry DCM Stir at −20 ° C. for 20 minutes before adding an equivalent amount of NIS and 1 drop of trifluoromethanesulfonic acid (TfOH).

  Stirring was continued at −20 ° C. until completion of the reaction was confirmed by TLC (˜1 hour) before 400 μl of triethylamine was added. Glycosylation products were obtained by concentration on silica gel (under vacuum) and flash chromatography.

General procedure for quaternary reaction of eel Methanol, methanol-THF (variable ratio) or chloroform containing acid (1 eq), amine (1 eq), carbonyl compound (1 eq) and isocyanide (1 eq) Was transferred to a reaction vial (final concentration: 0.1-0.5M). When D-gluconic acid was used as the acid, it was added in a solid state. In the case of bis-acid, bis-amine, bis-aldehyde or bis-isocyanide, the amount was 0.5 equivalent. The mixture was stirred or shaken at room temperature for 1 hour to 5 days. The progress of the reaction was monitored by TLC. The residue obtained by concentrating the mixture was purified by flash chromatography, or completely dried under high vacuum conditions and then purified by flash chromatography after direct overacetylation.

General procedure for acetylation of the hydroxyl group The corresponding alcohol was dissolved in DCM-pyridine (15: 1 v / v, 0.15 M) containing DMAP (0.42 mol%). Acetic anhydride (2 equivalents per hydroxyl group) was added and the resulting mixture was stirred overnight at room temperature. The mixture was poured into ice-cooled 0.5M hydrochloric acid and then extracted with chloroform. The organic phase was separated, washed with a salt solution, cold 0.5M hydrochloric acid (x2) and dried (magnesium sulfate). The resulting solution was filtered and concentrated. The residue was purified by flash chromatography (hexane-ethyl acetate gradient elution) to give a pure product.

General Procedure for Deacetylation / Debenzoylation of Zemprini A solution of anhydrous methanol (0.1M) containing acetate / benzoate was added to a solution of methanol (1.35M, 0.2-0.6 equiv) containing sodium methoxide. Processed. The mixture was stirred at room temperature for 1-3 hours (monitoring by TLC). Adding acidic resin AG ^(TM) -50 W-X8 (hydrogen ion form), the pH was adjusted to 6-7 and then the resulting mixture filtered and the resin was washed with methanol. The collected filtrate and washed product were concentrated under vacuum and completely dried to obtain a polyol product.

General Procedure for Deprotection of Benzyl Ether via Hydrogenolysis To a solution of benzyl ether protected compound (0.03 mmol) in methanol or ethanol (2 ml), 5% palladium / carbon or charcoal (30 mg or excess) 20% palladium hydroxide supported on) was added. The mixture was placed in a miniclave (Büch AG, Worcester / Switzerland) and stirring was continued for 2-10 hours under a hydrogen atmosphere (50 psi). Alternatively, hydrogen gas was introduced into the mixture for 1 hour and stirring was continued for 1-5 days at room temperature under 1 atmosphere of hydrogen. The reaction was monitored by TLC (ethyl acetate or methyl cyanide-water 10: 1). The mixture was filtered and then washed with methanol or ethanol. The filtrate was subjected to concentration and drying at high vacuum, confirmed by ¹ H NMR, lyophilized and used directly for sulfonation.

Polyol obtained by methylation drying of hydroxyl group was dissolved in anhydrous DMF (0.04M) in the presence of argon gas, and sodium hydride (60% suspension in mineral oil as a medium, hydroxyl group) at room temperature for 1 hour 1.2 equivalents per minute). Iodomethane (1.2 equivalents per hydroxyl group) was added and stirred overnight. Methanol was added and the mixture was concentrated using silica and purified by flash chromatography.

General Procedure for the Cycloaddition Reaction of Wisgen Sulfated sugar azide was dissolved in water (0.75 M) and then a solution of t-butanol (0.9 M, 1 eq) containing acetylene was added. To this mixture was added a solution of copper (II) sulfate (0.3 M solution in water, 5 mol%) and a solution of sodium ascorbate (1 M solution in water, 20 mol%). added. The mixture was shaken overnight at room temperature and column chromatography (1 × 18 cm, 50: 2: 1, 20: 2: 1 to 10: 2: 1 gradient elution of ethyl acetate-methanol-water) To obtain the corresponding triazole product.

Example 1: PG2038
Step a: Methyl 3,4,6-tri-O-acetyl-2-O-benzyl-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-β -D-glucopyranoside methyl 2,3,6-tri-O-benzyl-β-D-glucopyranoside [17] (150 mg, 322 μmol), methyl 3,4,6-tri-O-acetyl-2-O-benzyl- General NIS glycosylation was performed using 95 mg (422 μmol) NIS with 1-thio-β-D-galactopyranoside [18] (151 mg, 354 μmol) and 200 mg trimolecular sieve. Flash chromatography (gradient elution from 20:80 to 25:75 ethyl acetate: hexanes) gave 274 mg of partially deacetylated material. To this mixture was added 10 ml DCM, 200 μl acetic anhydride, 200 μl triethylamine and 2 mg DMAP, the resulting solution was stirred for 1 hour, concentrated and flash chromatography (25: 75- (Gradient elution of 30:70 ethyl acetate: hexane) gave 180 mg (66%) of the title compound as a colorless glass. ¹ H nmr (400 MHz, CDC1 ₃ ) δ: 7.05-7.35 (m, 20H, 4 × Ph), 5.74 (d, 1H, J _1,2 = 4.0, H1 ^II ), 5.29 (dd, J _3,4 = 3.2, J _4,5 = 1.2, H4 ^II ), 5.23 (dd, 1H, J _2,3 = 10.8, H3 ^II ), 4.92 (d, 1H, J _gem = 12.0, PhCH ₂ ), 4.85 (d, 1H, J _gem = 10.8, PhCH ₂ ), 4.55-4.69 (m, 4H, PhCH ₂ ), 4.42 (AB, 1H, J _gem = 12.0, PhCH ₂ ), 4.40 (AB, 1H, PhCH ₂ ), 4.32 ( d, 1H, J _1,2 = 7.6, H1 ^I ), 4.10 (dt, 1H, J _5,6 = 6.8, H5 ^II ), 3.88-3.96 (m, H, H5 ^I + H6 ^I ), 3.82 (dd , 1H, J _gem = 11.1, H6 ^II ), 3.70-3.76 (m, 4H, H2 × H6 ^I + H2 ^II + H3 ^I ), 3.56 (s, 3H, OMe), 3.5-3.6 (m, 1H, H4 ^I ), 3.45 (dd, 1H, J _2,3 = 9.0, H2 ^I ), 2.02 (s, 3H, Ac), 1.93 (s, 3H, Ac), 1.88 (s, 3H, Ac). ¹³ C nmr (100 MHz, CDCl ₃ ) δ: 170.17, 170.06, 169.87, 138.82, 138.20, 138.13, 137.68, 128.33, 128.25, 128.19, 128.04, 127.61, 127.57, 127.49, 127.46, 127.01, 126.39, 104.42, 97.12, 84.49, 82.26 , 74.46, 74.27, 73.69, 73.31, 73.28, 73.02, 69.47, 69.17, 68.35, 66.60, 61.62, 56.96, 20.69, 20.63, 20.58.

Step b: Methyl 3,4,6-tri-O-acetyl-α-D-galactopyranosyl- (1 → 4) -β-D-glucopyranoside Perlman's catalyst (20 mg) and 20 μl acetic acid Methyl 3,4,6-tri-O-acetyl-2-O-benzyl-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-β-D- It was added to 10 ml of methanol solution containing glucopyranoside. Hydrogen gas was fed in at 3 degree of vacuum and the suspension was stirred for 3 days. After filtration and concentration and co-concentration with toluene, the residue was applied to flash chromatography (gradient elution from 100: 0 to 100: 3 ethyl acetate: methane) to yield 47 mg (91%) of the title compound. Got. ¹ H nmr (400 MHz, CDC1 ₃ ) δ: 5.39 (br d, J _3,4 = 3.0, H4 ^II ), 5.32 (d, 1H, J _1,2 = 3.8, H1 ^II ), 5.10 (dd, 1H , J _2,3 = 10.6, H3 ^II ), 4.38 (br t, 1H, J _5,6 = 6.8, H5 ^II ), 4.20 (d, 1H, J _1,2 = 7.8, H1 ^I ), 4.09 (app d ( AB X), 2H, J _5,6 = 6.5, H6 ^II ), 4.00 (dd, 1H, H2 ^II ), 3.92 (dd, 1H, J _5,6A = 1.7, J _gem = 12.2, H6A ^I ) , 3.80 (dd, 1H, J _5,6B = 4.8, H6B ^I ), 3.62 (dis t, 1H, J _2.3 to _3,4 = 9.1, H3 ^I ), 3.56 (part obs t, 1H, J _3.4 to _{4 , 5} = 9.3, H4 ^I ), 3.53 (s, 3H, OMe), 3.42 (ddd, 1H, J _4,5 = 9.4, H5 ^I ), 3.22 (dd, 1H, J _2,3 = 9.1, H2 ^I ), 2.10 (s, 3H, AcO), 2.05 (s, 3H, AcO), 2.00 (s, 3H, AcO).

Step c: Methyl 2-O-sulfo-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-sulfo-β-D-glucopyranoside, tetrasodium salt (PG2038)
The above disaccharide (32.2 mg, 0.667 mmol) was applied to a standard sulfonation and deacetylation process to give the title compound as a white foam (4.0 mg, 7.8%, 96% purity, CE : 7.18 minutes). ¹ H NMR (D ₂ O, 400 MHz): 5.473 (d, 1H, J _1II-2II = 3.6, H1 ^II ), 4.833 (d, 1H, J _II-2I = 2.8, H1 ^I ), 4.60 (overlap with water H3 ^I ), 4.551 (m, 1H, H2 ^I ), 4.306 (dd, J _2II-3II = 10.2, H2 ^II ), 4.17-4.06 (m, 4H, H4 ^I , H5 ^I and H6 ^I ), 3.902 (d, 1H, J _3II-4II = 3.6, H4 ^II ), 3.867 (dd, 1H, H3 ^II ), 3.616 (dd, 1H, J _6axII-6eqII = 12.0, J _5II-6axII = 7.2, H6ax ^II ), 3.564 (dd, 1H, J _5II-6eqII = 5.2, H6eq ^II ), 3.363 (dd, 1H, H5 ^II ), 3.343 (s, 3H, CH30).

Example 2: PG2046 and PG2047
Step a: 2-Azido-3,4,6-tri-O-benzoyl-2-deoxy-α-D-glucopyranosyl- (1 → 4) -1,6-anhydro-2-deoxy-3-O-benzyl -β-D-glucopyranose 3,4,6-tri-O-acetyl-2-azido-2-deoxy-D-glucopyranosyltrichloroacetimidate [19] (201 mg, 0.453 mmol) and 6-anhydro A solution of 1,2-DCE (5 ml) containing -2-azido-3-O-benzyl-2-deoxy-β-D-glucopyranose [20] (84 mg, 0.302 mmol) was activated molecular sieve ( Stirring was performed in the presence of 300 mg powder) and under an argon atmosphere (at room temperature for 30 minutes). The mixture was cooled (−20 ° C.) with stirring (10 minutes), then TBDMSOTf (21 μl, 0.091 mmol) was added dropwise and stirring was continued further (−20 ° C., 10 minutes). ). Triethylamine (100 μl) was added and the resulting mixture was filtered and then concentrated. The residue was added to an aqueous test solution (ethyl acetate) and flash chromatography (10-40% ethyl acetate / hexane) to give a pale yellow oil (130 mg). The residue was co-concentrated (2 × 10 ml of methyl cyanide) and deacetylated according to the method of Zemprini. This product was added to an aqueous sample analysis (ethyl acetate) to give a clear oil (98 mg). The residue was co-concentrated (2 × 10 ml methyl cyanide). Benzoyl chloride (210 μl, 1.81 mmol) was added to a solution of crude product (up to 0.302 mmol) and 1,2-DCE (3 ml) containing pyridine (2 ml) and the combined mixture was stirred ( Room temperature overnight). The mixture was cooled (0 ° C.) and methanol (2 ml) was added with continued stirring (0 ° C. → room temperature, 2 minutes) before the solvent was evaporated and co-evaporated (toluene). The residue was added to an aqueous test solution (ethyl acetate) and subjected to flash chromatography (10-30% ethyl acetate / hexane) to give two compounds.

First, the title compound was a clear foam (101 mg, 46%, 3 steps). ¹ H NMR (400 MHz, CDC1 ₃ ) δ: 3.11 (s, 1H; H-2 ^I ), 3.41 (dd, 1H, J _1,2 3.7 J _2,3 10.7 Hz; H-6 ^I ), 3.61 ( s, H; H-3 ^I ), 3.39 (s, 1H; H-4 ^I ), 4.05 (d, 1H, J _6.6 7.3 Hz; H-6 ^I ), 4.41-4.49 (m, 2H; H-6 ^II ), 4.55, 4.68 (AB quadrupole, J _{A, B} 11.9 Hz; CH ₂ Ph), 4.79 (ddd, 1H, J _4,5 10.3, J _5,6 2.9, 5.9 Hz; H-5 ^II ) , 4.91 (br d, 1H J _5,6 5.5 Hz; H-5 ^I ), 5.08 (d, 1H, J _1,2 3.6 Hz; H-1 ^II ), 5.51 (dd, 1H, J _3,4 9.5 , J _4,5 10.2 H-4 ^II ), 5.60 (s, 1H, H-1 ^I ), 6.10 (dd, 1H J _2,3 10.7, J _3,4 9.3 Hz; H-3 ^II ), 7.29- . 7.55, 7.89-8.03 (2 m, 20H; ArH) 13 C NMR (100 MHz, CDC1 3) δ: 58.74, 61.47, 63.64, 69.19, 69.49, 70.52, 73.17, 74.62, 78.12, 79.57 (11 C; C -2 ^I -6 ^I , C2 ^II -6 ^II , CH ₂ Ph), 100.71, 101.16 (2 C; C-1 ^I , C-1 ^II ), 128.10, 128.42, 128.57, 128.61, 128.64, 128.81, 128.88, 129.15, 129.86, 129.90, 130.06, 130.17, 133.43, 133.53, 133.74, 137.48 (Ar), 165.61, 165.62, 166.26 (3 C; C = O).

Next, 2-azido-3,4,6-tri-O-benzoyl-2-deoxy-β-D-glucopyranosyl- (1 → 4) -1,6-anhydro-2-deoxy-3-O-benzyl -β-D-Glucopyranose was a clear oil (27 mg, 12%, 3 steps). ¹ H NMR (400 MHz, CDC1 ₃ ) δ: 3.19 (s, 1H; H-2 ^I ), 3.74 (dd, 1H, J _5,6 6.2 J _6,6 7.1 Hz; H-6 ^I ), 3.79- 3.88 (m, 2H; H-2 ^II , H-3 ^I ), 3.88 (ddd, J _4,5 9.2, J _5,6 3.1, 4.7 Hz; H-5 ^II ), 3.95 (br s, 1H; H -4 ^I ), 4.10 (d, 1H, J _6,6 7.3 Hz; H-6 ^I ), 4.34 (dd, 1H; J _5,6 4.9, J _6,6 12.2 Hz; H-6 ^II ), 4.50 (dd, 1H, J _5,6 3.1, J _6,6 12.3Hz, H-6 ^II ), 4.52, 4.59 (AB quadrupole, J _{A, B} 12.0 Hz; CH ₂ Ph), 4.65 (d, 1H , J _{l, 2} 7.9 Hz; H-1 ^II ), 4.69 (br d, 1H, J _5,6 5.5 Hz; H-5 ^I ), 5.44 (t, 1H, J _{2,3 = 3,4} 9.7 Hz ; H-3 ^II ), 5.49 (br s, 1H; H-1 ^I ), 5.51 (t, qH, J _{3,4 = 4,5} 9.6 Hz; H-4 ^II ), 7.23-7.50, 7.84-7.96 (2m, 20H; ArH).

Step b: 2-Deoxy-2-sulfamide-α-D-glucopyranosyl- (1 → 4) -1,6-anhydro-2-deoxy-2-sulfamide-3-O-benzyl-β-D-glucopyranose, Disodium salt (PG2046)
2-Azido-3,4,6-tri-O-benzoyl-2-deoxy-α-D-glucopyranosyl- (1 → 4) -1,6-anhydro-2-azido-2-deoxy-3-O- 2: 1 methanol: ethyl acetate (7 mL) containing a mixture of benzyl-β-D-glucopyranose (127 μmol), Perlman's catalyst (11 mg), and ammonium formate (300 mg) was added to an argon atmosphere until the end of TLC. Under, heated at 65 ° C. The mixture was cooled to room temperature and then filtered (0.2 μm) and concentrated. The crude amine was purified by SPE (300 mg C18 aqueous cartridge equilibrated with methanol: water = 5: 95 and eluted with a gradient of methanol: water = 5: 95 to 100: 0) to give 53 mg of diamine. (58%) was obtained. Without further purification, DMF (5 ml), SO ₃ .Me ₃ N (41 mg, 295 μmol) and NaHCO ₃ (40 mg, 475 μmol) were added to the diamine. The mixture was heated at 60 ° C. for 1 hour and cooled to room temperature before being quenched with ice and sodium carbonate (saturated, aqueous). This suspension was stored at −18 ° C. overnight before the sample was filtered. The filtrate was concentrated to dryness. Water (1 ml) and sodium hydroxide (250 μl, 1M) are added and the resulting solution is stirred overnight and then applied directly to a SEC column (general procedure) to give 22 mg of the title compound. Obtained (28% over 3 steps). ¹ H NMR (400 MHz, D ₂ 0, suppressed solvent) δ: 7.35-7.21 (m, 5H, ArH), 5.43 (br s, 1H, H1 ^I ), 5.18 (d, 1H, J _1-2 = 3.6, H ^II ), 4.72-4.69 ¹ (m, 1H, H5 ^I ), 4.54-4.52 ¹ (m, 2H, ArCH ₂ ), 4.05 (d, 1H, J _gem = 7.9, H6A ^I ), 3.85 (br s, 1H, H3 ^I ), 3.76-3.58 (m, 5H), 3.51 (dd, 1H, J _2-3 = 10.4, J _3-4 = 9.1, H3 ^II ), 3.34 (t, 1H, J _{3- . 4 ~ 3-5 = 9.2, H4} II), 3.23 (br s, 1H, H2 I), 3.12 (dd, 1H, H2 II) 13 C NMR (100 MHz, CDCl 3) δ: 133.3, 124.8, 124.6 , 124.4, 96.9, 95.1, 72.7, 71.5, 70.8, 68.3, 68.2, 67.2, 66.0, 61.0, 56.6, 54.0, 49.8.

Step c: 2-Deoxy-2-sulfamide-α-D-glucopyranosyl- (1 → 4) -1,6-anhydro-2-deoxy-2-sulfamide-β-D-glucopyranose, disodium salt (PG2047)
2-deoxy-2-sulfamide-α-D-glucopyranosyl- (1 → 4) -1,6-anhydro-2-deoxy-2-sulfamide-3-O-benzyl-β-D-glucopyranose, disodium salt To purified water (2 ml) containing (12.9 mg, 20.8 μmol) and Pearlman's catalyst (5 mg), 50 psi of hydrogen gas was bubbled overnight. The mixture was filtered and lyophilized to give 10.7 mg of the title compound (98%). ¹ H NMR (400 MHz, D ₂ 0) δ: 5.47 (br s, 1H, H1 ^I ), 5.20 (d, 1H, J _1-2 = 3.5, H1 ^II ), 4.68 (br d, 1H, J _{5 -4 = 5.5, H5), 4.07} (d, 1H, J gem = 7.6, H6A I), 3.98 (br s, 1H, H3 I), 3.75-3.64 (m, 4H), 3.52 (t, 1H, J _2-3 to _3-4 = 9.3, H3 ^II ), 3.34 (t, 1H, J _3-4 to _4-5 = 9.3, H4 ^II ), 3.13 (obs.dd ² , 1H, H2 ^II ), 3.11 ( br s, 1H, H2 ^I ).
^{1 The} effect of solvent suppression signal was observed.

² Partially unclear due to H2 ^I effects.

Example 3: PG2039 and PG2037
Step a: Methyl 3,4-di-O-acetyl-2,6-di-O-benzyl-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl -β-D-glucopyranoside methyl 2,3,6-methyl-tri-O-benzyl-β-D-glucopyranoside (287 mg; 618 μmol), 302 mg (618 μmol) of ethyl 3,4-di-O-acetyl-2, 6-O-dibenzyl-1-thio-β-D-galactopyranoside [21] and 700 mg of 3 molecular sieves were used to perform general NIS glycosylation using 181 mg (803 μmol, 1.3 eq) NIS. went. Flash chromatography (2.5 × 20 cm, 1: 5 to 1: 3 gradient elution of ethyl acetate: hexanes) produced the title compound as a clear gum (176 mg, 32%). ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.40-7.12 (m, 25H, Ph), 5.818 (d, 1H, J _III-2II = 3.6, H1 ^II ), 5.481 (d, 1H, J _4II-3II = 3.2, H4 ^II ), 5.309 (dd, 1H, J _3II-2II = 10.8, J _3II-4II = 3.2, H3 ^II ), 4.980 (d, 1H, J _gem = 11.6, a-PhCH ₂ ), 4.904 ( d, 1H, J _gem = 11.2, b-PhCH ₂ ), 4.748 (d, 1H, J _gem = 11.6, a-PhCH ₂ ), 4.67-4.57 (m, 3H, b-PhCH ₂ and c-PhCH ₂ ) , 4.479 (d, 1H, J _gem = 12.8, d-PhCH ₂ ), 4.443 (d, 1H, J _gem = 12.8, d-PhCH ₂ ), 4.415 (d, 1H, J _gem = 11.6, e-PhCH ₂ ), 4.360 (d, 1H, J _II-2I = 8.0, H1 ^I ), 4.201 (d, J _gem = 12.6, e-PhCH ₂ ), 4.153 (t, J _5II-6axII = 7.2, J _5II-6eqII = 6.0, H5 ^II ), 4.072 (t, 1H, J _4I-3I = 9.0, J _4I-5I = 9.0, H4 ^I ), 3.858 (dd, 1H, J _2II-3II = 10.8, J _2II-1II = 3.6, H2 ^II ), 3.82-3.76 (m, 3H, H3I, H6ax ^I and H6 eq ^I ), 3.597 (s, 3H, OMe), 3.62-3.57 (m, 1H, H5 ^I ), 3.507 (t, 1H, J _2I-3I = 8.4, J _2I-II = 8.0, H2 ^I ), 3.347 (dd, J _6eqII-6axII = 9.2, J _6eqII-5II = 6.0, H6eq ^II ), 3.291 (dd, 1H, J _6axII-6eqII = 9.2, J _6axII-5II = 7.2, H6ax ^II ), 1.958 (s, 3H, OAc), 1.930 ( ¹³ C NMR (100 MHz, CDCl ₃ , 100 MHz): 169.94 (CO), 169.78 (CO), 138.88, 138.35, 138.24, 137.73 and 137.64 (5x ipso-Ph), 128.25, 128.22 , 128.21,128.15, 128.12, 128.00, 127.80, 127.57, 127.52, 127.46, 127.41, 127.37, 126.93, 126.37, 104.40, 97.16, 84.63, 82.31, 74.42, 74.30, 73.70, 73.25, 73.16, 73.14, 72.98, 69.73, 69.07 , 68.89, 67.64, 67.50, 56.86, 20.71, 20.55.

Step b: Methyl 3,4-di-O-acetyl-α-D-galactopyranosyl- (1 → 4) -β-D-glucopyranoside According to the standard debenzylation method, methyl 3,4-di Protection of -O-acetyl-2,6-di-O-benzyl-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-β-D-glucopyranoside Release of the title compound as a clear powder (42 mg, 97%). ¹ H NMR (D ₂ 0, 400 MHz) δ: 5.394 (d, 1H, J _1II-2II = 3.6, H1 ^II ), 5.294 (d, 1H, J _4II-3II = 3.2, H4 ^II ), 4.953 (dd , 1H, J _3II-2II = 10.4, J _3II-4II = 3.2, H3 ^II ), 4.229 (d, 1H, J _II-2I = 8.4, H1 ^I ), 4.080 (t, 1H, J _5II-6axII = 6.4 , J _5II-6eqII = 6.0, H5 ^II ), 3.965 (dd, 1H, J _2II-3II = 10.4, J _2II-1II = 3.6, H2 ^II ), 3.803 (dd, 1H, J _6eq1-6eqI = 12.0, J _6eqI-5I = 1.6, H6eq ^I ), 3.67-3.59 (m, 2H, H6ax ^I and H3 ^I ), 3.54-3.40 (m, 4H, H4I, H5 ^I and H6 ^II ), 3.407 (s, 3H, OMe) , 3.134 (dd, 1H, J 2I-3I = 9.2, J 2I-II = 8.4, H2 I), 2.012 (s, 3H, OAc), 1.909 (s, 3H, OAc). 13 C NMR (D 2 0 , 100 MHz): 173.57, 173.47, 103.20, 99.71, 77.12, 76.30, 74.57, 73.09, 70.89, 69.94, 69.04, 66.45, 60.82, 60.18, 57.31, 20.34, 20.09.

Step c: Methyl 2,6-di-O-sulfo-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-sulfo-β-D-glucopyranoside, pentasodium salt (PG2039)
From 42 mg (95.4 μmol) of methyl 3,4-di-O-acetyl-α-D-galactopyranosyl- (1 → 4) -β-D-glucopyranoside by standard sulfonation / deacetylation, The title compound was obtained as a white powder (14.8 mg, 18%, CE: 6.12 min). ¹ H NMR (D ₂ 0, 400 MHz): 5.404 (d, 1H, J _1II-2II = 3.6, H1 ^II ), 4.756 (d, 1H, J _1I-2I = 3.6, H1 ^I ), 4.60 (with water Duplicate, 1H, H3 ^I ), 4.448 (dd, 1H, J _2I-3I = 3.2, H2 ^I ), 4.296 (dd, 1H, J _2II-3II = 10.0, H2 ^II ), 4.23-4.00 (m, 7H, H6 ^I , H5 ^I , H6 ^II , H4 ^I and H5 ^II ), _3.958 (dd, 1H, J _3II-4II = 3.6, J _4II-5II = 0.8, H4 ^II ), 3.930 (dd, 1H, H3 ^II ), 3.367 (s, 3H, CH ₃ 0).

Step d: Methyl 2,6-di-O-benzyl-3,4-di-O-methyl-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl -β-D-glucopyranoside by standard deacetylation and methylation, methyl 3,4-di-O-acetyl-2,6-di-O-benzyl-α-D-galactopyranosyl- (1 → 4) -2,3,6-Tri-O-benzyl-β-D-glucopyranoside (72 mg, 80.8 μmol) gave the title compound as a clear gum (62.7 mg, 93%). ¹ H NMR (CDCl ₃ , 400 MHz): 7.35-7.08 (m, 25H, Ph), 5.717 (d, 1H, J _1II-2II , 3.6, H1 ^II ), 4.856 (d, J _gem = 11.2, a- PhCH ₂ ), 4.843 (d, 1H, J _gem = 10.8, b-PhCH ₂ ), 4.695 (d, 2H, J _gem = 12.0, a-PhCH ₂ and c-PhCH ₂ ), 4.631 (d, 1H, J _gem = 12.4, d-PhCH ₂ ), 4.571 (d, 1H, J _gem = 10.8, b-PhCH ₂ ), 4.500 (d, 1H, J _gem = 12.4, d-PhCH ₂ ), 4.450 (d, 1H, J _gem = 11.6, c-PhCH ₂ ), 4.433 (d, 1H, J _gem = 11.2, e-PhCH ₂ ), 4.359 (d, 1H, J _gem = 11.2, e-PhCH ₂ ), 4.303 (d, 1H , J _1I-2I = 7.6, H1 ^I ), 3.949 (t, 1H, J _4I-3I = 9.0, J _4I-5I = 9.0, H4 ^I ), 3.871 (dd, 1H, J _5II-6axII = 7.2, J _5II-6eqII = 6.4, H5 ^II ), 3.791 (dd, 1H, J _2II-3II = 10.4, J _2II-1II = 3.6, H2 ^II ), 3.77-3.68 (m, 4H, H4 ^II , H3 ^I , H6ax ^I And H6eq ^I )), 3.59-3.53 (m, 2H, H5 ^I and H6 ^II ), 3.551 (s, 3H, OMe), 3.51-3.40 (m, 3H, H3 ^II , H6 ^II and H2 ^I ), 3.492 ( s, 3H, OMe), 3.433 (s, 3H, OMe).

Step e: Methyl 3,4-di-O-methyl-α-D-galactopyranosyl- (1 → 4) -β-D-glucopyranoside by standard debenzylation, methyl 2,6-di-O -Benzyl-3,4-di-O-methyl-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-β-D-glucopyranoside (62.7 mg, 75.1 μmol) gave the title compound as a clear gum (28 mg, 97%). ¹ H NMR (D ₂ O, 400 MHz): 5.232 (d, 1H, J _1II-2II = 4.4, H1 ^II ), _4.217 (d, 1H, J _1I-2I = 8.0, H1 ^I ), 3.83-3.75 ( m, 3H, H4 ^II , H5 ^II and H6 ^I ), 3.682 (dd, 1H, J _2II-3II = 10.4, J _2II-1II = 4.4, H2 ^II ), 3.64-3.52 (m, 4H, H6, H3 ^II And H6 ^II ), 3.47-3.38 (m, 3H, H4 ^I , H5 ^I and H3 ^II ), 3.400 (s, 3H, OMe), 3.340 (s, 6H, 2xOMe), 3.117 (dd, 1H, J _{2I- . 3I = 9.6, J 2I-} 1I = 8.0, H2 I) 13 C NMR (D 2 O, 100 MHz): 103.20, 99.64, 79.35, 76.92, 76.34, 75.35, 74.65, 73.06, 72.03, 68.00, 61.02, 60.89 , 60.86, 57.30, 56.94.

Step f: Methyl 3,4-di-O-methyl-2,6-di-O-sulfo-α-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-sulfo -β-D-glucopyranoside, pentasodium salt (PG2037)
The title compound is obtained from methyl 3,4-di-O-methyl-α-D-galactopyranosyl- (1 → 4) -β-D-glucopyranoside (28 mg, 72.8 μmol) by standard sulfonation. (3.2 mg, 4.9%). ¹ H NMR (400 MHz, D ₂ O): 5.357 (d, 1H, J _1II-2II = 3.2, H1 ^II ), 4.766 (d, 1H, J _II-2I = 3.6, H1 ^I ), 4.60 (with water Duplicate, 1H, H3 ^I ), 4.455 (dd, 1H, J _2I-3I = 2.8, H2 ^I ), 4.304 (dd, 1H, J _2II-3II = 10.0, H2 ^II ), 4.22-3.99 (m, 5H, H5 ^I , H4 ^I and H5 ^II ), 4.002 (d, 2H, J _5II-6II = 6.8, H6 ^II ), 3.886 (d, 1H, J _3II-4II = 3.2, H4 ^II ), 3.667 (dd, 1H, H3 ^II ), 3.398 (s, 3H, CH ₃ 0), 3.367 (s, 3H, CH ₃ 0), 3.356 (s, 3H, CH ₃ 0).

Example 4: PG2053 and PG2042
Methyl 4-aryl-2,3-di-O-sulfo-α-L-rhamnoside, disodium salt (PG2053)
The title compound was obtained as a clear powder from rhamnopyranoside [22] via general alkylation (utilizing aryl bromide) and deprotection steps, followed by general sulfonation. CE t _m = 10.48 min. ¹ H NMR (400 MHz, D ₂ O) δ 1.19 (d, 3H, J _5,6 = 6.4 Hz; H-6), 3.26 (s, 3H; OMe); 3.29-3.40 (m, 1H; H-4), 3.59-3.67 (m, 1H; H-5), 4.00-4.05, 4.18-4.22 (2m, 2H; OCH ₂ ), 4.41-4.42 (m, 1H; H-3 ), 4.63-4.64 (m, 2H; H-2), 4.83 (s, 1H; 5.07-5.21 (m, 2H; = CH ₂ ), 5.76-5.88 (m, 1H; = CH).
When a small amount (1 equivalent) of SO ₃ .trimethylamine was used, only methyl 4-O-aryl-2-O-sulfo-α-L-rhamnoside, sodium salt (PG2042) was obtained. CE t _m > 25.00 min. ¹ H NMR (400 MHz, D ₂ O) δ 1.19 (d, 3H, J _5,6 6.4 Hz; H-6), 3.16 (t, 1H, J _3,4 3.1, J _4,5 9.7 Hz; H-4), 3.25 (s, 3H; OMe), 3.54-3.58 (m, 1H; H-5), 3.76 (dd, 1H, J _2,3 9.7 Hz; H-3) , 4.03-4.18 (m, 2H; OCH ₂ ), 4.34-4.35 (m, 1H; H-2), 4.80 (s, 1H; H-1), 5.08-5.22 (m, 2H; = CH ₂ ), 5.78-5.88 (m, 1H; = CH).

Example 5: PG2024
Methyl 4-O-benzyl-2,3-di-O-sulfo-α-L-rhamnoside, disodium salt (PG2024)
The title compound is obtained from methyl 2,3-O-isopropylidene-α-L-rhamnopyranoside via general alkylation (utilizing benzyl bromide) and deprotection steps, followed by general sulfonation. Obtained as a transparent powder. CE t _m = 10.82 min. ¹ H NMR (400 MHz, D ₂ O) δ 1.00 (d, 3H, J _5,6 6.8Hz; H-6), 3.23 (s, 3H; OMe); 3.78-3.80 ( m, 1H; H-4), 3.88-3.94 (m, 1H; H-5), 4.41-4.43 (m, 1H; H-2), 4.52-4.56 (m, 2H; H-3), 4.54, 4.78 (AB quadrupole, J _{A, B} 12.0 Hz; CH ₂ Ph), 4.90 (dd, 1H, J _1,2 1.2 Hz; H-1), 7.20-7.36 (m, 5H; ArH).

Example 6: PG2054
Step a: Methyl 4-O-benzyl-.alpha.-L-rhamnoside methyl 2,3-O-isopropylidene-.alpha.-L-rhamnopyranoside (200mg, 920μmol), benzoyl chloride (193 mg, 1.38 mmol) and triethylamine (364μl, 2.76 mmol) in DCM (10 ml) was stirred overnight. The resulting suspension (triethylamine / hydrochloric acid precipitate) is diluted with DCM (50 ml) and washed with sodium bicarbonate (saturated solution), water and brine before being dried (magnesium sulfate) and Concentrated. The residue was dissolved in 50 ml of 1: 1 methyl cyanide: water mixture and p-TsOH (10 mg) was added. The resulting solution was stirred until the reaction was complete (TCL, ˜4 hours), concentrated and then flash chromatographed (1: 1 ethyl acetate: hexanes) to give 165 mg of a clear solid. The title compound was obtained (64% over two steps). ¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.03-8.00 (m, 2H, H ortho), 7.55 (tt, 1H, J _Hp-Hm = 7.5, J _Hp-Ho = 1.3, H pillar), 7.43- 7.39 (m, 2H, H meta), 5.09 (dis t, 1H, J _4-3 to _4-5 = 9.3, H4), 4.73 (br s, 1H, H1), 4.01-3.97 (m, 2H, H2 + H3), 3.91 (dq, 1H, J _4-5 = 9.7, J _5-6 = 6.4, H5), 3.53-3.46 (br s, 2H, OH), 3.38 (s, 3H, OMe), 1.25 ( ^{. d, 3H, H6) 13} C NMR (100 MHz, CDCl 3) δ: 167.1, 133.3, 129.8, 129.5, 128.3, 100.6, 75.7, 70.8, 70.1, 65.7, 55.0, 17.4.

Step b: Methyl 4-O-benzoyl-2,3-di-O-sulfo-α-L-rhamnoside, disodium salt (PG2054)
The title compound was obtained as a clear powder from methyl 4-O-benzoyl-α-L-rhamnoside via general sulfonation. CE t _m = 11.14 min. ¹ H NMR (400 MHz, D ₂ O) δ 1.14 (d, 3H, J _5,6 6.3Hz; H-6), 3.33 (s, 3H; OMe), 3.99-4.07 ( m, 1H; H-5), 4.66-4.73 (m, 2H; H-2, -3), 4.95 (d, 1H, J _1,2 1.4 Hz; H-1), 5.04 (t, 1H, J _{3,4 = 4,5} 9.6Hz; H-4), 7.35-7.41, 7.53-7.55, 7.92-7.93 (3m, 5H; Ph).

Example 7: PG2041
Step a: 4,6-O- benzylidene-1,2-dihydro -D- glucaric tri -O- acetyl -D- glucaric (1.7g, 6.25mmol), acetic acid (50 [mu] l) and Pd (OH) ₂ / C ( 100 mg) of methanol (15 ml) was stirred vigorously overnight in the presence of hydrogen gas (1 atm). The mixture was filtered, the solvent was evaporated, and the residue obtained was flash chromatographed (10-50% ethyl acetate / hexane) to give a clear oily tri-O-acetyl-1, 2-Dihydro-D-glucal was obtained. The residue was co-concentrated (2 × 10 ml of methyl cyanide) and subjected to the deacetylation of semprene to give 1,2-dihydro-D-glucal as a clear oil (825 mg, 89 %). The residue was co-concentrated (2 × 10 ml methyl cyanide).

P-TsOH.H ₂ O (50 mg) was added to a solution of 1,2-dihydro-D-glucar (495 mg, 3.34 mmol) and DMF (5 ml) containing α, α-dimethoxytoluene (735 μl, 5.01 mmol). The mixture was stirred (60 ° C., 1 hour). Triethylamine (100 μl) was poured and the solvent was evaporated. When the residue was subjected to flash chromatography (0 to 5% methanol / chloroform), a transparent needle-like title compound was obtained (503 mg, 64%). ¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.72-2.01 (m, 2H; H-2), 3.27-3.33 (m, 1H; H-5), 3.41 (dd, J3,4 8.8, J5,6 9.1Hz; H-4), 3.49-3.56 (m, 1H; H-3), 3.67 (t, 1H, J5,6 = 6,6 10.3Hz; H-6), 3.81-3.87, 3.93-3.98 ( 2m, 2H; H-1), 4.25 (dd, 1H; J5,6 4.9Hz; H-6), 5.53 (s, 1H; CHPh), 7.23-7.52 (m, 5H, CHPh). ¹³ C NMR ( 100 MHz, CDCl ₃ ) δ: 33.47 (C-2); 66.46, 69.07, 69.64, 71.32 (4C; C-1, -4, -5, -6), 84.14, (C-3), 102.16 (CHPh ), 126.43, 128.55, 129.34, 137.50 (4C; Ph).

Step b: 3-O-benzyl-4,6-di-O-sulfo-1,2-dihydro-D-glucal, disodium salt (PG2041)
By subjecting 4,6-O-benzylidene-1,2-dihydro-D-glucal to alkylation (using benzyl bromide), deprotection step, and general sulfonation, the title compound is obtained. Obtained as a transparent powder. CE t _m = 15.40 min. ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.48-1.53, 1.97-2.03 (2m, 2H; H-2), 3.30-3.71 (m, 1H; H-1), 3.52- 3.57 (m, 1H; H-5), 3.60-3.66 (m, 1H; H-3), 3.78-3.83 (m, 1H; H-1), 3.97 (dd, 1H, J _5,6 8.0, J _6,6 11.4Hz; H-6), 3.98 (t, 1H, J _{3,4 = 4,5} 8.9Hz; H-4), 4.34 (dd, 1H, J _5,6 2.3Hz; H-6) , 4.52-4.67 (m, 2H; CH ₂ Ph), 7.21-7.36 (m, 5H; Ph).

Example 8: PG2030
Step a: 1,6- Anhydro -3-O-methyl-β-D-glucopyranose p-toluenesulfonyl chloride (790 mg, 4.14 mmol) was added to 3-O-methyl-β-D-glucopyranose (804 mg, 4.14). In addition to pyrimidine (10 ml) containing a cooled (0 ° C.) suspension of mmol), the resulting mixture was stirred (0 ° C. → room temperature, 1.5 h). Acetic anhydride (1.5 ml, 15 mmol) and N, N-dimethylaminopyridine (50 mg) were added, and stirring was continued (room temperature, 4 hours). The mixture was cooled (0 ° C.), methanol (3 ml) was added, and stirring was continued until the solvent was evaporated (10 minutes). The remaining oil was dissolved (with ethyl acetate) and subjected to analysis to give the tosylate as a pale yellow oil (1.93 g). Ethanol (20 ml) containing a mixture of crude tosylate (1.93 g) and sodium hydroxide (1.0 M 20 ml, 20 mmol) was heated (80 ° C., 1 hour). The mixture was neutralized with acetic acid and solvent evaporation and co-evaporation (toluene) was performed. The crude residue was treated with pyridine (50 mg), acetic anhydride (5 ml) and N, N-dimethylaminopyridine (50 mg) and the resulting mixture was stirred (room temperature, overnight). The mixture was treated with ice water (10 ml) and stirring was continued (3 hours at room temperature) before analysis (ethyl acetate). The resulting residue was subjected to flash chromatography (20-50% ethyl acetate / hexane) to find 2,4-di-O-acetyl-1,6-anhydro-3-O-methyl-β-. D-glucopyranose (a) and 1,2,4-tri-O-acetyl-3-O-methyl-6-O-tosyl-α-D-glucopyranose (b) (in a 3: 1 ratio) An inseparable mixture containing was obtained as a pale yellow oil (466 mg). This ratio was determined based on the distribution of H-1 and 3-OMe that can be read from the ¹ H NMR spectrum. Partial ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.32 (s, 3H; OMe b), 3.45 (s, 3H; OMe a); 5.22 (br s, 1H; H-1 b), 5.42 (br s , 1H; H-1 a). A mixture of the two compounds (456 mg) was deacetylated and the resulting residue was subjected to flash chromatography (0-5% methanol / ethyl acetate) to give the title compound as a clear oil As a product (162 mg, 33% over 3 steps). ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.27-3.30 (m, 1H; H-3), 3.38 (s, 3H; OMe), 3.57-3.59 (m, 1H; H-2), 3.63-3.65 ( m, 1H; H-4), 3.70 (dd, 1H, J _5,6 = 5.6, J _6,6 = 7.2 Hz; H-6), 4.06 (d, 1H, J _6,6 = 7.2 Hz; H -6), 4.48-4.51 (m, 1H; H-5), 4.39 (br s, 1H; H-1).

Step b: 1,6- anhydro -4-O-benzyl-3-O-methyl-β-D-glucopyranose 1,6-anhydro-3-O-methyl-β-D-glucopyranose (155 mg, 0.88 mmol ) And dibutyltin oxide (241 mg, 0.97 mmol) in toluene (18 ml) was heated under reflux (water was removed azeotropically) until the volume of the solution was half the original. The mixture was cooled (80 ° C.), then benzyl bromide (104 μl, 0.88 mmol) and tetrabutylammonium bromide (567 mg, 1.76 mmol) were added and stirring was continued (overnight). The mixture was treated with methanol (2 ml) and water (1 ml) with continued stirring (10 minutes) before evaporation of the solvent. The residue was analyzed (ethyl acetate) and subjected to flash chromatography (20-60% ethyl acetate / hexanes) to give two compounds.

First, the title compound was obtained as a clear oil (94 mg, 40%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.58 (d, 1H, J _{2, OH} 6.4Hz; OH), 3.32-3.43 (m, 5H; H-3, H-4, OMe), 3.52-3.57 ( m, 1H; H-2), 3.68-3.72, 4.01-4.04 (2m, 2H; H-6), 4.55-4.58 (m, 1H; H-5), 4.64 (s, 2H; CH ₂ Ph), 5.39-5.40 (m, 1H; H-1), 7.28-7.36 (m, 5H; ArH).
1,6-Anhydro-2-O-benzyl-3-O-methyl-β-D-glucopyranose was then obtained as a clear oil (91 mg, 39%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.90 (br s, 1H; OH), 3.31-3.34 (m, 4H; H-2, OMe), 3.36-3.38 (m, 1H; H-3), 3.58 (br s, 1H; H-4), 3.68 (dd, 1H, J _5,6 6.0Hz, J _6,6 7.2 Hz; H-6), 4.08 (dd, 1H J _5,6 0.8Hz, J _{6 , 6} 7.2 Hz; H-6), 4.47-4.49 (m, 1H; H-5), 4.56, 4.62 (AB quadrupole, J _{A, B} 12.0Hz; CH ₂ Ph), 5.40-5.41 (m, 1H; H-1), 7.26-7.36 (m, 5H; ArH).

Step c: 1,6-anhydro-4-O-benzyl-3-O-methyl-2-O-sulfo-β-D-glucopyranose, sodium salt (PG2030)
1,6-Anhydro-4-O-benzyl-3-O-methyl-β-D-glucopyranose (82 mg, 0.32 mmol) was sulfonated according to conventional methods and then flash chromatographed (50/2/1 → 10/2/1 ethyl acetate / methanol / water) and SEC gave the title compound as a pale yellow powder (70 mg, 60%). CE t _m = 5.62 min. ¹ H NMR (400 MHz, D ₂ O) δ 3.19 (s, 3 H; OCH ₃ ); 3.43-3.45 (m, 1H; H-4), 3.52-3.53 (m, 1H ; H-3), 3.57 (dd, 1H, J _5,6 = 5.9Hz, J _6,6 = 7.8Hz; H-6), 3.82 (dd, 1H, J _5,6 = 1.1Hz, J _{6, 6} = 7.8Hz; H-6), 3.97-3.99 (m, 1H; H-5), 4.59-4.61 (m, 3H; H-2, CH ₂ Ph), 5.41 (br s, 1H; H-1 ), 7.22-7.34 (m, 5H; ArH).

Example 9: PG2012 and PG2013
Step a: N-benzyl -N- after finishing the general procedure for the (cyclohexyl acetamide) -1,2,3,4 -O- acetyl -D- Gurukuronamido Ugi reaction, D- gluconic acid (0.950 g, 4.89 mmol) and the following three reagents: benzylamine (2M in methanol, 2.45 ml, 4.89 mmol), formaldehyde (2M in methanol, 2.45 ml, 4.89 mmol) and cyclohexyl isocyanide (1M Of methanol solution, 2.45 ml, 4.89 mmol) was placed in a reaction vessel and the mixture was then stirred at room temperature for 19 hours. The volatile component was removed under reduced pressure and dried under pressure to obtain yellow foam N-benzyl-N- (cyclohexylacetamide) -D-glucuronamide.

After finishing the general procedure for acetylation, the crude eel reaction product described above was peracetylated and subjected to flash chromatography (gradient elution of hexane: ethyl acetate 2: 1 to 1: 1). The title compound was obtained as a 1.929 g, 66% pale yellow foam (two steps, Rf = 0.37, hexane: ethyl acetate 1: 1). Due to the presence of anomers and rotamers, ¹ H NMR (CDCl ₃ , 400 MHz) became very complex. Even after heating to 55 ° C., the spectrum was not simplified. However, each set of rotamers at 100 ° C. pyridine-d ₆ had a simpler structure, so two anomers could be clearly observed (α: β = 69: 31). ¹ H NMR (CDCl ₃ , 400 MHz, 25 ° C): 7.41-7.14 (m, 5H, Ph), 6.337 (d, 0.39H, J = 3.6), 6.300 (d, 0.29H, J = 3.6), 5.969 (br d, 0.52H, J = 8), 5.823 (br d, 0.09H, J = 8.4), 5.66-5.41 (m, 2.25H), 5.28-5.09 (m, 1.3H), 4.92-4.58 (m , 2.25H), 4.411 (d, J = 10) and 4.395 (d, J = 14, 0.55H), 4.271 (d, 0.11H, J = 9.6), 4.219 (d, 0.09H, J = 17.2), 4.125 (d, 0.17H, J = 14), 4.098 (d, 0.17H, J = 14.4), 3.994 (d, J = 15.2) and 3.963 (d, J = 14.8, 0.89H), 3.82-3.59 (m , 2.04H), 2.190, 2.111, 2.038, 2.033, 2.025, 2.023, 2.016, 2.014, 2.008, 1.998, 1.983, 1.944 and 1.927 (all singles, 12H, Ac), 1.89-1.55 (m, cyclohexyl-CH ₂ ), 1.41-0.83 (m, 5H, cyclohexyl-CH ₂ ). ¹ H NMR (CDCl ₃ , 400 MHz, 55 ° C): 7.38-7.16 (m, 5H, Ph), 6.336 (d, J = 3.2,) And 6.153 (d, J = 3.2, 0.7H), 5.939 (br d, 0.6H, J = 6.8), 5.721 (br d, 0.2H, J = 7.2), 5.65-5.55 (m, 1.3H), 5.52 -5.41 (m, 1H,), 5.28-5. 10 (m, 1.4H), 4.85-4.58 (m, 2.4H), 4.48-4.40 (m, 0.6H), 4.318 (d, 0.1H, J = 9.2), 4.205 (d, 0.1H, J = 17.6), 4.02-3 .93 (m, 0.9H), 3.84-3.62 (m, 2.2H), 2.179, 2.090, 2.021, 2.012, 2.001, 1.989, 1.983, 1.975, 1.968, 1.959 and 1.935 (all singles, 12H, Ac), 1.88-1.56 (m, 5H, cyclohexyl-CH ₂ ), 1.42-0.88 (m, 5H, cyclohexyl-CH ₂ ). ¹ H NMR (pyridine-d ₆ , 400 MHz, δ7.22, 100 ° C) is generally Only the typical sugar protons, the remaining signal (other than the acetate singlet) was complex and showed a large clumpy signal. α-anomer; 6.672 (d, J = 3.6, glu-H1), 5.456 (dd, J = 9.6, 3.6, glu-H2); β-anomer; 6.206 (d, J = 8.0, glu-H1), 5.741 (t, J = 9.2, glu-H4 or H5), 5.515 (dd, J = 8.8, 8.0, glu-H2).

Step b: N-benzyl-N- (cyclohexylacetamide) -1,2,3,4-tetra-O-sulfo-α-D-glucuronamide, tetrasodium salt (PG2012) and N-benzyl-N- ( (Cyclohexylacetamide) -1,2,3-tri-O-sulfo-α-D-glucuronamide, trisodium salt (PG2013)
The tetraacetate salt (0.441 g, 0.747 mmol) described above was deacetylated by the general procedure for deacetylation to give pale yellow glassy N-benzyl-N- (cyclohexylacetamide) -D- Glucuronamide was obtained (0.316 g, 100%).

The above-described tetrol (0.257 g, 0.608 mmol) was sulfonated (19 hours using sulfur trioxide pyridine complex) by the general procedure for sulfonation. The residue was co-concentrated with toluene and then flash chromatographed [2.5 × 20 cm, ethyl acetate, methyl cyanide, methyl cyanide-triethylamine (10: 1), methyl cyanide-triethylamine-water (110: 2:11)]. These fractions were divided in two by TLC and CE. A less polar sample was purified again by flash chromatography, LH20 (× 2) and ion exchange chromatography to give a white fluffy powder PG2013 trisulfate (19.3 mg) after lyophilization. , 4.4%). ¹ H NMR (D ₂ O, 400 MHz): Two rotamers in a ratio of 56:44. Major rotamers, δ 7.36-7.11 (m, 5H, Ph), 5.946 (d, 1H, J = 3.2, H1) , 4.894 (d, 1H, J = 9.6, H5), 4.748 (d, 1H, J = 16, a-CH ₂ ), 4.685 (d, 1H, J = 16, a-CH ₂ ), 4.502 (t, 1H, J = 10.4, 9.6, H3), 4.306 (dd, 1H, J = 9.6, 3.6, H2), 4.005 (t, 1H, J = 9.6, 8.8, H4), 3.869 (s, 2H, b-CH ₂ ), 3.42-3.32 (m, 1H, cyclohexyl-CHN), 1.64-1.36 (m, cyclohexyl-CH ₂ ), 1.20-0.92 (m, cyclohexyl-CH ₂ ); minor rotamers, 7.36-7.11 (n, 5H, Ph), 5.905 (d, 1H, J = 3.2, H1), 4.578 (d, 1H, J = 10, H5), 4.523 (s, 2H, c-CH ₂ ), 4.478 (t, J = 10.4 , 9.6, H3), 4.321 (d, 1H, J = 17.6, d-CH ₂ ), 4.281 (dd, 1H, J = 9.6, 3.2, H2), 4.039 (t, 1H, J = 9.6, 9.2, H4 ), 3.900 (d, 1H, J = 17.6, d-CH ₂ ), 3.42-3.32 (m, 1H, cyclohexyl-CHN), 1.64-1.36 (m, 5H, cyclohexyl-CH ₂ ), 1.20-0.92 . (m, 5H, cyclohexyl ^{_{-CH 2) 13 C NMR (D}} 2 O, 100MHz, no reference values): due to two rotamers 169.96 (Amid-CON), 169.75 (Amid-CON), 168.91 (Amid-CON), 168.85 (Amid-CON), 135.45 (ipso-Ph), 135.20 (ipso-Ph), 129.16, 129.07, 128.47, 128.32, 128.20 and 128.13 (Meta-, Ortho- and Phala-Ph), 95.67 and 95.65 (glu-C1), 77.84 and 77.75 (glu-C3), 73.76 and 73.71 (glu-C2), 70.66 and 70.18 (glu-C4), 69.23 and 68.70 (glu-C5), 52.87 (a-CH ₂ ), 51.12 (c-CH ₂ ), 50.32 (d-CH ₂ ), 50.02 (b-CH ₂ ), 49.25 and 49.00 (cyclohexyl-CHN), 31.89 and 31.86, 25.08 and 25.05, 24.47 and 24.38 (cyclohexyl-CH ₂ ). ES-LRMS (+ ve, m / z): C ₂₁ H ₂₇ N ₂ Na ₃ O ₁₆ S ₃ is considered 728.02, but is actually 751 (M + Na ⁺ ), 729 (M + H ⁺ ); ES-HRMS (+ ve, m / z): M + Na ⁺ , C ₂₁ H ₂₇ N ₂ Na ₄ O ₁₆ S ₃ is considered 751.0113, actually, 750.1087; M + H a _{^{+; C 21 H 28 N 2}} Na 3 O 16 S 3 is considered 729.0294 In fact, it was 729.0242.

The polar part was purified by LH20 (× 2) column and ion exchange column to obtain gray powder tetrasulfate PG2012 (7.6 mg, 1.5%) after lyophilization. ¹ H NMR (D ₂ O, 400 MHz): two rotamers in a molar ratio of 70:30. Major rotamers, δ 7.34-7.16 (m, 5H, Ph), 5.954 (d, 1H, J = 3.6, H1 ), 5.235 (d, 1H, J = 9.6, H5), 4.904 (d, 1H, J = 15.6, a-CH ₂ ), 4.67-4.57 (overlap with water, 1H, H3), 4.536 (t, 1H, J = 9.6, 8.8, H4), 4.466 (d, 1H, J = 15.6, a-CH ₂ ), 4.394 (dd, 1H, J = 9.8, 3.4, H2), 3.927 (d, 1H, J = 16.8, b-CH ₂ ), 3.749 (d, 1H, J = 16.8, b-CH ₂ ), 3.30-3.20 (m, 1H, cyclohexyl-CHN), 1.65-1.35 (m, 5H, cyclohexyl-CH ₂ ), 1.18 -0.92 (m, 5H, cyclohexyl-CH ₂ ); minor rotamer, 7.34-7.16 (m, 5H, Ph), 5.912 (d, 1H, J = 3.4, H1), 4.77-4.72 (m, 2H, H5 And H3 or H4), 4.689 (d, 1H, J = 15.2, c-CH ₂ ), 4.67-4.56 (overlap with water, 1H, H4 or H3), 4.373 (dd, 1H, J = 9.8, 3.4, H2 ), 4.256 (d, 1H, J = 18.4, d-CH ₂ ), 4.215 (d, 1H, J = 15.2, c-CH ₂ ), 3.936 (d, 1H, J = 18.4, d-CH ₂ ), 3.36-3.26 (m, 1H, cyclohexyl-CHN), 1.65-1.35 (m, 5H, cyclohexyl-CH ₂ ), 1.18-0.92 (m, 5H, Cyclohexyl-CH ₂ ). ¹³ C NMR (D ₂ O, 100 MHz, no reference): Major Rotamer, 172.35 (Amid-CON), 171.69 (Amid-CON), 137.22 (ipso-Ph), 131.71 and 131.58 (Meta- and Ortho-Ph), 131.11 (Hala-Ph), 97.68 (glu-C1), 78.08 (glu-C4), 77.60 (glu-C3), 76.43 (glu-C2), 70.10 (glu-C5) , 55.81 (a-CH ₂ ), 53.17 (b-CH ₂ ), 51.77 (cyclohexyl-CHN), 34.17 (cyclohexyl-CH ₂ ), 27.56 (cyclohexyl-CH ₂ ), 27.13 (cyclohexyl-CH ₂ ); minor Rotamer (only general peak appears), 97.65 (glu-C1), 78.22 (glu-C4), 77.77 (glu-C3), 76.33 (glu-C2), 71.01 (glu-C5), 53.26 (d- CH ₂ ), 53.79 (c-CH ₂ ), 52.10 (cyclohexyl-CHN), 34.28 (cyclohexyl-CH ₂ ), 27.52 (cyclohexyl-CH ₂ ), 27.08 (cyclohexyl-CH ₂ ) .ES-MS (+ ve, m / z): C ₂₁ H ₂₆ N ₂ Na ₄ O ₁₉ S ₄ is considered to be 829.96, but was actually 853 (M + Na ⁺ ) and 831 (M + H ⁺ ). ES-HRMS (+ ve, m / z): M + Na ⁺ , C ₂₁ H ₂₆ N ₂ Na ₅ O ₁₉ S ₄ is considered 852.9501, but is actually 852.9334; M + H ⁺ , C ₂₁ H ₂₇ N ₂ Na ₄ O ₁₉ S ₄ is considered to be 830.9682, but was actually 830.9635.

Example 10: PG2064
Step a: 2- (N-acetyl-N-cyclohexyl) amino-N- (methyl-2,3,4-tri-O-benzyl-6-deoxy-α-D-mannopyranose-6-yl) acetamide After completing the general procedure for the eel reaction, four reagents, namely acetic acid (2 M methanol solution, 60 μl, 119 μmol), cyclohexylamine (2 M methanol solution, 60 μl, 119 μmol), formaldehyde (2 M methanol solution) , 60 μl, 119 μmol), and 4 ml of each solution of methyl 2,3,4-tri-O-benzyl-6-deoxy-6-isocyano-α-D-mannopyranoside (0.721 M chloroform, 150 μl, 108 μmol) The reaction mixture was then stirred at 60 ° C. for 19 hours. Volatile components were removed under reduced pressure and purified by flash chromatography (gradient elution with 4: 1 to 1: 4 hexane: ethyl acetate) to yield 42 mg, 60% (Rf = 0.49, ethyl acetate). The title compound was obtained as a clear gum. ¹ H NMR (CDCl ₃ , 400 MHz): two rotamers in the ratio 72:28, δ 7.38-7.26 (m, 15H, 3 × C ₆ H ₅ ), 6.933 (t, 72% × 1H, J = 4.4 , NH in major rotomer), 6.357 (t, 28% × 1H, J = 5.8, NH in minor rotomer), 4.91-4.41 (m, 7H, sugar-H1 and 3 × PhCH ₂ ), 4.04- 3.42 (m, 9H, Sugar-H2-6, NCH ₂ CO and Cyclohexyl-CH), 3.305 (s, 72% × 3H, CH ₃ 0 in major rotamer), 3.266 (s, 28% × 3H, minor・ CH ₃ in Rotamer, 2.067 (s, 72% × 3H, CH ₃ CO in Major Rotamer), 2.012 (s, 28% × 3H, CH ₃ CO in Major Rotamer), 1.85-1.00 (m, 10H, cyclohexyl -CH _2).

Step b: 2- (N-acetyl-N-cyclohexyl) amino-N- (methyl 6-deoxy-2,3,4-tri-O-sulfo-α-D-mannopyranose-6-yl) acetamide, Trisodium salt (PG2064)
After finishing the general procedure for deprotection of benzyl ether, methanol (2 ml) containing a mixture of the above tribenzyl ether (42 mg, 0.065 mmol), 20% palladium on activated charcoal (22 mg) was added to hydrogen Stir for 10 hours at 50 psi under atmosphere. In this way, a clear gum triol intermediate was obtained. By the general procedure for sulfonation, the triol was sulfonated (at 60 ° C. for 19 hours, the sulfur trioxide complex was obtained) and the crude product was concentrated. The residue was purified by continuous SEC (Bio Gel P-2 followed by LH20). The purified product was converted to the sodium salt through an ion exchange column to give the title compound as a white fluffy powder after lyophilization (two steps, 3.1 mg, 7.0%). ¹ H NMR (D ₂ O, 2.05 acetone state reference, 400 MHz): 63:37 ratio of two rotamers, δ 4.844 (s, 1H, sugar-H1), 4.678 (s, 1H, sugar- H2), 4.51-4.45 (m, 1H, Sugar-H3), 4.240 (t, 37% × 1H, J = 9.6, Sugar-H4 in minor rotamer), 4.214 (t, 63% × 1H, J = 9.6, Sugars in major rotamers-H4), 3.980 (s, 37% x 2H, COCH ₂ N in minor rotamers), 3.825 (s, 63% x 2H, COCH ₂ N in major rotamers), 3.78-3.59 (m, 3H, sugar-H5, single sugar-H6 and cyclohexyl-CH), 3.31-3.17 (m, 4H, single sugar-H6 and CH ₃ O [3.253, s, 3H]), 2.060 (s, 67% × 3H, CH ₃ CO in major rotormer), 1.900 (s, 33% × 3H, CH ₃ CO in minor rotorer), 1.70-0.88 (m, 10H, cyclohexyl-CH ₂ ) .

Example 11: PG2068
Step a
After completing the general procedure for the eel reaction, monomethyl succinate (15.7 mg, 0.119 mmol) and three reagents: ethylamine (2M methanol solution, 60 μl, 119 μmol), formaldehyde (2M methanol solution) 2 ml of each solution of methyl 2,3,4-tri-O-benzyl-6-deoxy-6-isocyano-α-D-mannopyranoside (0.721 M chloroform, 150 μl, 108 μmol), 60 μl, 119 μmol) In a reaction vessel and then the mixture was stirred at room temperature for 19 hours. The volatile components were removed under reduced pressure and purified by flash chromatography (gradient elution with 1: 1 to 1: 4 hexane: ethyl acetate followed by elution with ethyl acetate), clear. A quality gummy refined was obtained (46.8 mg, 65%). ¹ H NMR (CDCl ₃ , 400 MHz): two rotamers in a ratio of 73:27. Δ 7.38-7.25 (m, 15H, Ph), 6.598 (t, 73% × 1H, J = 6, NH), 6.529 (t, 27% × 1H, J = 6, NH), 4.93-4.61 (m, 7H, Sugar-H1 and 3 × PhCH ₂ ), 4.14-3.24 (m, 16H, Sugar 6 × H, NCH ₂ CO, 2 × CH ₃ O [single line at 3.660 and 3.301, 73%; 3.648 and 3.276, 27%] and ethyl-CH ₂ ), 2.70-2.42 (m, 4H, COCH ₂ CH ₂ CO), 1.138 (t, 73% × 3H, J = 7, ethyl-CH ₃ ), 1.014 (t, 27% × 3H, J = 7, ethyl-CH ₃ ). ¹³ C (100 MHz, CDC1 ₃ , δ 77.0): Major rotamer , 173.32, 171.73, 168.94, 138.20, 138.17, 138.09, 128.22, 128.18, 128.10, 127.76, 127.63, 127.49, 127.46, 98.94, 79.97, 75.38, 75.01, 74.84, 73.01, 72.01, 70.10, 54.63, 51.63, 49.84 , 39.63, 28.99, 27.24, 13.45. Minor rotamers (only non-overlapping peaks only), 173.26, 171.54, 168.01, 128.27, 127.69, 127.55, 98.98, 79.85, 75.14, 74.40, 72.86, 71.92, 69.97, 54.74, 50.91, 49.72, 42.10, 28.90, 27.83, 12.29.

Step b (PG2068)
After finishing the general procedure for deprotection of benzyl ether, methanol (3 ml) containing a mixture of the above tribenzyl ether (46.8 mg, 0.0706 mmol), 20% palladium on activated charcoal (30 mg) was added. Stir for 2 hours at 50 psi under hydrogen atmosphere. In this way, a clear gum triol intermediate was obtained. The triol was sulfonated by the general procedure for sulfonation (sulphur trioxide complex was obtained at 60 ° C. for 19 hours). The residue was dissolved in 1M sodium hydroxide (3 ml, 0.16M). The mixture was stirred at room temperature overnight and concentrated under reduced pressure. The residue was purified by continuous SEC (Bio Gel P-2 followed by LH20). This purified product was converted to a sodium salt through an ion exchange column to obtain PG2068 as a white powder (two steps, 4.9 mg, 9.8%). ¹ H NMR (D ₂ O, 2.05 acetone state reference, 400 MHz): two rotamers in a ratio of 70:30, δ 4.86-4.84 (m, 1H, sugar-H1), 4.69-4.67 (m, 1H, Sugar-H2), 4.50-4.46 (m, 1H, Sugar-H3), 4.28-4.19 (m, 1H, Sugar-H4), 4.072 (s, 30% × 2H, COCH ₂ N in minor rotomer 2H), 3.987 (d, 35% × 2H, J = 16.8, 1H for COCH ₂ N for major rotamers), 3.815 (d, 35% × 2H, J = 16.8, 1H for COCH ₂ N for major rotamers ), 3.79-3.69 (m, 2H, sugar-H5 and one sugar-H6), 3.44-3.18 (m, 6H, one sugar-H6, ethyl-CH ₂ and CH ₃ 0 [3.245, s, 3H] ), 2.633 (t, J = 6.8) and 2.45-2.37 (m, total 4H, COCH ₂ CH ₂ CO ₂ ), 1.054 (t, 70% × 3H, J = 7.2, CH ₃ 0), 0.906 (t, 30% × 3H, J = 7.2, CH ₃ 0).

Example 12: PG2075
Step a
3-Chlorophenylacetic acid (223 mg, 1.307 mmol) was dissolved in methyl cyanide (3 ml). Ammonia solution (28%, 0.26 ml, 3.8 mmol) was added. The mixture was stirred for a while and then concentrated in vacuo. The residue was suspended in methyl cyanide (3 ml), then the white solid obtained by filtration was washed with methyl cyanide and lyophilized to give ammonium 3-chlorophenylacetate ( 0.195g, 80%).

  After completing the general procedure for the eel reaction, the ammonium salt (22.5 mg, 0.120 mmol) described above and two reagents: formaldehyde (2M in methanol, 60 μl, 119 μmol) and methyl 2-isocyanoethyl Each solution of 2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside (0.762 M chloroform, 157 μl, 120 μmol) was placed in a 2 ml reaction vessel and the mixture was then The mixture was stirred at room temperature for 19 hours.

Volatile components were removed under reduced pressure and purified by flash chromatography to give a clear gum-like purified product (34.8 mg, 37%). ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.39-7.06 (m, 24H, 4 × C ₆ H ₅ and 1 × C ₆ H ₄ ), 6.731 (t, 1H, J = 6.0, NH), 4. (d, 1H, J = 10.8, a-CH ₂ ), 4.844 (d, 1H, J = 2.0, sugar-H1), 4.770 (d, 1H, J = 12.4, b-CH ₂ ), 4.723 (d, 1H, J = 12.4, b-CH ₂ ), 4.640 (s, 2H, c-CH ₂ ), 4.589 (d, 1H, J = 11.6, d-CH ₂ ), 4.535 (d, 1H, J = 11.6, d-CH ₂ ), 4.505 (d, 1H, J = 10.8, a-CH ₂ ), 4.446 (d, 1H, J = 14.8, e-CH ₂ ), 4.362 (d, 1H, J = 14.8, e- _{. CH 2), 3.90-3.51 (m} , 12H), 3.38-3.29 (m, 1H) 13 C (100 MHz, CDC1 3, δ77.0): 169.67, 166.81, 138.27, 138.11, 137.70, 135.16, 134.31, 129.82, 129.40, 128.34, 128.31, 128.01, 127.85, 127.78, 127.70, 127.68, 127.60, 127.53, 127.46, 98.90, 79.96, 75.08, 75.04, 74.71, 73.58, 72.73, 72.19, 72.13, 69.78, 68.49, 63.17, 0.25 39.27.

Step b (PG2075)
After finishing the general procedure for deprotection of benzyl ether, methanol (2 ml) containing a mixture of the above tetrabenzyl ether (34.8 mg, 0.0439 mmol), 20% palladium on activated charcoal (26 mg) was added. Stir for 2 hours at 50 psi under hydrogen atmosphere. By doing so, a tetrol intermediate product of a transparent gum was obtained. The tetrol was sulfonated by the general procedure for sulfonation. The residue was purified by SEC (Bio Gel P-2). This purified product was converted to a sodium salt by passing through an ion exchange column, and PG2075 as a white fluffy powder was obtained after lyophilization (two steps, 10.6 mg, 28%). ¹ H NMR (D ₂ O, 2.05 acetone state reference, 400 MHz): δ 7.30-7.11 (m, 4H, Ar), 5.00-4.97 (m, 1H, sugar-H1), 4.72-4.28 (m , 3H, sugar-H2, H3 and H4), 4.21-3.30 (m, 11H, sugar-H5, H6 and 4 × CH ₂ ).

Example 13: PG2014
Step a
After finishing the general procedure for the eel reaction, 2- (benzyl 3,4,6-tri-O-benzyl-α-D-mannopyranoside-2-yl) acetic acid (50 mg, 0.0835 mmol) and three Reagents: benzylamine (2M in methanol, 41.8 μl, 0.0835 mmol), formaldehyde (2M in methanol, 41.8 μl, 0.0835 mmol), and 2-isocyanoethyl 2,3,4,6-tetra-O— Each solution of benzyl-α-D-mannopyranoside (0.415 M in methanol, 201.4 μl, 0.0835 mmol) was placed in a 2 ml reaction vessel and the mixture was then stirred at room temperature for 19 hours. . A clear gummy refined was obtained (38.9 mg, 36%).

¹ H NMR (400 MHz): two rotamers in a ratio of 66:34 around the amide single bond of CO-NH, δ 7.40-7.05 (m, 45H, 9 × C ₆ H ₅ ), 6.66 (t, 0.66H , J = 5.6, CONH-Major Rotamer) and 6.40 (t, 0.34H, J = 5.4, CONH-Minor Rotamer), 5.104 (d, 0.66H, J = 1.2, H1 ^I -Major Rotamer) and 5.046 (s, 0.34H, H1 ^I -minor rotamer), 4.86-4.20 (m, 21H), 3.97-3.48 (m, 16H), 3.36 (q, 0.66H, J = 5.6, major rotamer) and 3.26 ( q, 0.34H, J = 5.6, minor rotormer).

Step b (PG2014)
After finishing the general procedure for deprotection of benzyl ether, ethanol (4 ml) containing a mixture of the above octabenzyl ether (35 mg, 0.0267 mmol), 20% palladium on activated charcoal (10 mg) was added to hydrogen. Stir for 2 hours at 50 psi under atmosphere. By doing so, a tetrol intermediate product of a transparent gum was obtained. This octol was sulfonated by a general procedure for sulfonation (sulfur trioxide complex was obtained at 60 ° C. for 19 hours). The residue was purified by SEC (Bio Gel P-2). This purified product was converted to a sodium salt through an ion exchange column to obtain PG2014 as a white powder (two steps, 16.6 mg, 44%). ¹ H NMR (D ₂ O, 400 MHz, complicated by two rotamers): δ 7.32-7.14 (m, 5H, Ph), 5.80-5.66 (m, 0.5H), 5.44-5.39 (m, 0.5 H), 5.04-4.96 (m, 1.5H), 4.80-4.20 (m, 9H, overlapping with water), 4.18-3.78 (m, 7.5H), 3.76-3.46 (m, 1.5H), 3.42-2.98 (m, 3.5H).

Example 14: PG2016
Step a
After completing the general procedure for the eel reaction, trans-1,4-diaminocyclohexane (6.3 mg, 0.055 mmol) and three reagents, namely 2- (methyl 2,3,4-tri-O— Benzyl-α-D-mannopyranoside-6-yl) acetic acid (0.91 M in methanol, 121 μl, 0.11 mmol), formaldehyde (2 M in methanol, 55 μl, 0.11 mmol), and cyclohexyl isocyanide (1 M in methanol, 110 μl, 0.11 mmol) of each solution was placed in a 2 ml reaction vessel and the mixture was then stirred at room temperature for 5 days. The volatile components were removed under reduced pressure and purified by flash chromatography (gradient elution with 2: 1 to 1: 4 hexane: ethyl acetate) to yield 33.0 mg, 43%, a clear gum (Rf = 0.24, DCM-methanol = 95: 5 or Rf = 0.48, methyl cyanide-ethyl acetate = 1: 1). ¹ H NMR (D ₂ O, 400 MHz, very complex due to multiple rotamers): δ 7.50-7.20 (m, 30H, Ph), 6.62 (br s, 6.48 (br s, 1.1H), 6.48 (br, s, 0.38H), 5.96 (br d, 0.26H, J = 8), 5.79 (br d, 0.26H, J = 10), 4.92-4.85 (m, 2H), 4.78-4.56 (m, 12H), 4.28-4.22 (m, 2.8H), 4.35-4.06 (m, 1.6H), 3.94-3.56 (m, 19.6H), 3.28 (s, 6H, OMe), 1.88-1.42 (m, 16H) , 1.36-1.00 (m, 12H).

Step b (PG2016)
After finishing the general procedure for deprotection of benzyl ether, ethanol (2.8 ml) containing a mixture of the above hexabenzyl ether (33 mg, 0.0235 mmol), 20% palladium on activated charcoal (65 mg), The mixture was stirred under a hydrogen atmosphere at 1 atm for 5 days. By doing so, a tetrol intermediate product of a transparent gum was obtained. This hexol was sulfonated by the general procedure for sulfonation. The residue was purified by continuous column chromatography (Bio Gel P-2 followed by SEC using an ion exchange column) to give PG2016 as a white powder (12.2 mg, 35%). ¹ H NMR (D ₂ O, 400 MHz): δ 4.97-4.92 (m, 2H, man-H1), 4.77-4.75 (m, 2H, man-H2), 4.58-4.52 (m, 2H, man-H3 ), 4.46-4.08 (m, 6H, including man-H4 at 4.46-4.36 and OCH ₂ CO), 3.98-3.80 (m, 8H, man-H5, man-H6 and 3.80-3.32 (m, 6H, including man-H6 at 3.80-3.64, and cyclohexyl-CH ₂ ), 3.318 (s, 6H, OMe), 1.82-1.34 (m, 18H, cyclohexyl-CH ₂ ), 1.26-1.00 (m, 10H, cyclohexyl-CH ₂ ). ES-MS (+ ve, m / z): C ₄₀ H ₆₂ N ₄ Na ₆ O ₃₄ S ₆ is thought to be 1472.10, but in fact 1495 (M + Na ⁺ ), 1473 (M + H ⁺ ) ES-HRMS (+ ve, m / z): M + Na ⁺ , C ₄₀ H ₆₂ N ₄ Na ₇ O ₃₄ S ₆ is considered 1495.853, but in fact, 1495.0957, and M + H ⁺ , C ₄₀ H ₆₃ N ₄ Na ₆ O ₃₄ S ₆ is considered to be 1473.1034, but was actually 1473.1082.

Example 15: PG2015
Step a
After finishing the general procedure for the Ugi reaction, 3,3-dimethylglutaric acid (7.1 mg, 0.0443 mmol) and three reagents, namely 3-aminopropyl 2,3,4,6-tetra-O -Benzyl-α-D-mannopyranoside (0.642 M in methanol, 138 μl, 0.0886 mmol), formaldehyde (2 M in methanol, 44.3 μl, 0.0886 mmol), and cyclohexyl isocyanide (1 M in methanol, 88.6 μl, 0.0886 mmol) Were placed in a 2 ml reaction vessel and the mixture was then stirred at room temperature for 5 days. Volatile components were removed under reduced pressure and purified by flash chromatography (gradient elution with 2: 1 to 1: 4 hexane: ethyl acetate), 32.3 mg, 46%, clear gum (Rf = 0.45, hexane-ethyl acetate = 1: 3). ¹ H NMR (CDCl ₃ , 400 MHz, very complex due to multiple rotamers): δ 7.38-7.21 (m, 40H, Ph), 7.003 (d, 0.41H, J = 7.7), 6.930 (br s , 0.21H), 6.726 (d, 0.4H, J = 8.4), 6.597 (d, 0.73H, J = 8.8), 6.487 (br s, 0.25H), 4.85-4.78 (m, 4H), 4.76-4.59 (m, 10H), 4.54-4.45 (m, 4H), 4.00-3.62 (m, 20H), 3.46-3.14 (m, 6H), 2.52-2.22 (m, 4H), 1.90-1.52 (m, 15H) , 1.34-1.00 (m, 15H).

Step b (PG2015)
After finishing the general procedure for deprotection of benzyl ether, ethanol (2.8 ml) containing a mixture of the above hexabenzyl ether (32.3 mg, 0.0202 mmol), 20% palladium on activated charcoal (41 mg) was added. The mixture was stirred in a hydrogen atmosphere at 1 atm for 5 days. By doing so, an octol intermediate product of transparent gum was obtained. This octol was sulfonated by the general procedure for sulfonation. The residue was purified by SEC (Bio Gel P-2) to give PG2015 as a white powder (12.6 mg, 37%). ¹ H NMR (D ₂ O, 400 MHz): δ 5.020 (d, 2H, J = 1.6, man-H1), 4.759 (br s, 2H, man-H2), 4.66-4.56 (m, 2H, man- H3, overlapping with water), 4.46-4.41 (m, 2H, man-H6), 4.265 (t, 2H, J = 9.6, 9.2, man-H4), 4.10-3.96 (m, 4H, man-H5 and man-H6), 4.05-3.14 (m, 14H, NCH ₂ CO, cyclohexyl-CH and NCH ₂ CH ₂ CH ₂ O), 2.50-2.11 (m, 4H, CCH ₂ CO), 1.84-1.42 (m, 14H , Cyclohexyl-CH ₂ and NCH ₂ CH ₂ CH ₂ O), 1.24-0.93 (m, 16H, cyclohexyl-CH ₂ and Me). ES-MS (+ ve, m / z): C ₄₁ H ₉₃ N ₁₁ O ₃₇ S ₇ (7 × SO ₃ NH ₄ ) is considered to be 1556, but was actually 1578 (M + Na ⁺ ) and 1556 (M + H ⁺ ). ES-HRMS (+ ve, m / z): M + H ⁺ , C ₄₁ H ₉₃ N ₁₁ O ₃₇ S ₇ is considered to be 1556.3857, but in actuality it was 1556.3783.

Example 16: PG2155
Ethyl 2,6-di-O-benzyl-3,4-di-O-sulfo-β-D-galactopyranoside, disodium salt (PG2155)
The title compound was obtained as a transparent powder from galactopyranoside [23] using a general sulfonation method. ¹ H NMR (400 MHz, D ₂ O): δ 1.10 (dd, 3H; CH ₂ CH ₃ ), 2.49-2.66 (m, 2H, CH ₂ CH ₃ ); 3.59 (dd, 1H, J _{1,22, 3 9.7Hz; H-2);} 3.65 (dd, 1H, J 5,6a 3.3, J 6a, 6b 11.0Hz; H-6a); 3.68 (dd, 1H, J 5,6b 4.0Hz; H-6b) ; 3.83 (m, 1H; H-5); 4.36 (dd, 1H, J _3,4 3.0Hz; H-3); 4.46 (s, 2H; CH ₂ Ph); 4.48 (d, 1H; H-1 ); 4.60, 4.75 (AB quadrupole, J 10.3Hz; CH ₂ Ph); 4.85 (dd, 1H; J _4,5 0.0Hz; H-4); 7.22-7.29, 7.37-7.39 (2m, 10H; ArH).

Example 17: PG2163
Step a: Methyl 4-O-aryl-6-azido-6-deoxy-2,3-di-O-isopropylidene -α-D-mannopyranoside methyl 6-azido-6-deoxy-α-D-mannopyranoside (311 mg , 1.419 mmol) was treated with (±) -camphor-10-sulfonic acid (16 mg, 0.0709 mmol, 5 mol%). The mixture was stirred at room temperature for 1 hour.

  TLC showed complete conversion to the title compound (Rf = 0.40, ethyl acetate-hexane = 17: 83). To this mixture was added saturated sodium carbonate (aqueous solution) to make it basic, then concentrated in vacuo. The residue was extracted with ethyl acetate (30 ml) and the ethyl acetate extract was washed with brine and dried (magnesium sulfate). The gum produced through filtration and concentration was once co-concentrated with toluene. The final clear gum was dissolved in anhydrous DMF (3.5 ml, 0.4 M) and stirred with sodium hydride (60% mineral oil dispersion, 163 mg, 4.257 mmol, 3 eq) for 1 hour. . Aryl bromide (360 μl, 4.257 mmol, 3 eq) was added and the resulting mixture was stirred at room temperature for 6 hours, treated with methanol (1 ml) and concentrated to dryness. The residue was purified by silica column chromatography (2.5 x 18 cm, eluting with 1:10 to 1: 6 ethyl acetate: hexanes) to give the title compound as a clear gum (0.281 mg, 2 steps). 66%).

¹ H NMR (CDCl ₃ , 400 MHz): 5.85 (m, 1H, aryl-2 '), 5.23 (ddd, 1H, J _2-3'trans = 17.2, J _3'-gem = 3.6, J _{1'- 3 '} = 1.6, H3' _trans ), 5.17-5.13 (m, 1H, H3 ' _cis ), 4.89 (s, 1H, H1), 4.35 (dddd, 1H, J _1'gem = 12.4, J _{1'-2 '} = 5.2, J = 1.6, H1'), 4.16 (dd, 1H, J _2-3 = 5.6, J _3-4 = 7.2, H3), 4.09 (d, 1H, H2), 4.05 (dddd, 1H, H1 '), 3.67 (ddd, J _4-5 = 10.4, J _5-6ax = 6.8, J _5-6eq = 2.4, H5), 3.48 (dd, 1H, J _6ax-6eq = 13.2, H6eq), 3.40 ( _{dd, 1H, J 6ax-6eq} = 13.2, J 5-6ax = 6.8, H6ax), 3.33 (dd, 1H, H5), 1.50 (s, 3H, Me), 1.30 (s, 3H, Me). 13 C NMR (CDCl ₃ , 100 MHz): 134.4, 117.1, 109.2, 98.0, 78.2, 76.2, 75.6, 71.5, 68.0, 54.9, 51.6, 27.8, 26.1.

Step b: Methyl 4-O-aryl-6-azido-6-deoxy-α-D-mannopyranoside methyl 4-O-aryl-6-azido-6-deoxy-2,3-di-O-isopropylidene-α -D-mannopyranoside (56 mg, 0.187 mmol) was dissolved in methyl cyanide-methanol-water (3 ml, 3 ml and 0.2 ml, respectively) and p-toluenesulfonic acid monohydrate (7 mg, 0.0374 mmol, 20 mol%). After the mixture was stirred at room temperature for 5 hours, triethylamine (0.4 ml) was added. The resulting mixture was concentrated and the residue was purified by column chromatography (1 × 18 cm silica, eluting with 1: 6 to 2: 1 ethyl acetate: hexanes) to give a clear waxy. A solid was obtained (34.8 mg, 72%). ¹ H NMR (CDCl ₃ , 400 MHz): 5.91 (m, 1H, aryl-H2 '), 5.28 (ddd, J _2'-3'trans = 16.8, J _{3'trans-3'cIs} = 3.0, J _{1 '-3'} = 1.6, aryl-H3'trans), 5.20 (ddd, J _2'-3'cIs = 10.0, J _{3'trans-3'cIs} = 3.0, J _{1'-3 '} = 1.6, aryl- H3'cIs), 4.72 (ddd, 1H, J _1-2 = 2.0, H1), 4.28 (dddd, 1H, J _gem = 12.4, J _{1'-2 '} = 5.6, J = 1.6, aryl-1') , 4.13 (dddd, 1H, aryl-1 '), 3.92 (dd, 1H, J _2-3 = 3.6, H2), 3.88 (dd, 1H, J _3-4 = 9.6, H3), 3.70 (ddd, 1H , J _4-5 = 9.6, J _5-6ax = 5.6, J _5-6eq = 2.8, H5), 3.54-3.44 (m, 3H, H4, H6ax and H6eq), 3.39 (s, 3H, MeO), 2.62 (br s, 2H, OH).

Step c: Methyl 4-O-aryl-6-azido-2-O-benzyl-6-deoxy-2,3-di-O-sulfonate-α-D-mannopyranoside disodium salt (PG2163)
Methyl 4-O-aryl-6-azido-6-deoxy-α-D-mannopyranoside was sulfonated according to standard procedures to give 55 mg (89%) of the title compound as a white powder. Rf = 0.20 (methyl cyanide: methanol: water = 10: 2: 1). ¹ H NMR (D ₂ O, 400 MHz): 5.88-5.76 (m, 1H, aryl-2 '), 5.20 (d, 1H, J _2'-3'trans = 17.2, aryl-3'trans), 5.12 (d, 1H, J _2'-3'cis = 10.00, aryl-3'cis), 4.91 (s, 1H, H1), 4.65 (br s, 1H, J _2-3 = 3.2, H2), 4.48 ( dd, 1H, J _2- = 3.2, J _3-4 = 9.2, H3), 4.21 (dd, 1H, J _gem = 11.6, J _{1'-2 '} = 5.6, aryl-1'), 4.00 (dd, 1H, J _{1'-2 '} = 6.4, aryl-1'), 3.71-3.67 (m, 1H, H5), 3.58 (dd, 1H, J _6eq-6ax = 13.6, J _5-6eq = 2.0, H6eq) , 3.58 (dd, 1H, J 4-5 = 9.6, H4), 3.46 (dd, 1H, J 6eq-6ax = 13.6, J 5-6ax = 5.6, H6ax), 3.30 (s, 3H, MeO). 13 C NMR (D ₂ O, 100 MHz, 49.05 ppm intrinsic methanol): 133.9, 119.2, 98.4, 76.1, 75.3, 74.2, 73.1, 70.7, 55.4, 50.8.

Example 18: PG2160, PG2161, and PG2173
Step a: Methyl 6-azido-6-deoxy-2,3-di-O-benzylidene -α-D-mannopyranoside methyl 6-azido-6-deoxy-α-D-mannopyranoside (1.011 g, 4.61 mmol) Dissolved in anhydrous DMF (9 ml) and acetonitrile (9 ml). Benzaldehyde dimethyl acetal (1.38 ml, 9.22 mmol, 2 eq) and (±) -camphor-10-sulfonic acid (214 mg, 0.922 mmol, 20 mol%) were added in this order. The resulting mixture was stirred indoors at 60 ° C. (external temperature) overnight and then the volatile components were removed on a rotary concentrator. The residue was passed through silica gel and column (2.5 × 20 cm silica, 10: 1, 9: 1, 7: 1, 5: 1, 4: 1, 3: 1 to 2: 1 hexane: ethyl acetate. Purified by gradient elution). These fractions were collected and divided into two groups. The small polar groups (Rf = 0.55 and 0.51, 3: 1, hexane: ethyl acetate) were 4-acetal mixtures, and the polar groups were mainly 4-OH products. These mixtures were combined and then concentrated. The residue was redissolved in dichloromethane (50 ml) and then stirred with 1M ammonium chloride solution (50 ml). The products with small polarity gradually disappeared and converted into polar products. However, after 40 minutes, no new conversion was observed. Therefore, the dichloromethane phase was separated and stirred with 0.5 M hydrochloric acid solution (50 ml) for a further 20 minutes. Thereafter, no change in TLC was observed. The DCM phase was separated and washed with brine-1M sodium hydroxide and then dried (magnesium sulfate). The dried DCM concentrate was filtered and concentrated, and the resulting residue was purified on the silica column described above to give the title compound as a clear gummy solid (0.543 g, Rf = 0.29, acetic acid). Ethyl-hexane = 1: 3). ¹ H NMR (CDCl ₃ , 400 MHz): Two benzylidene epimers in a 1: 1 ratio. 7.49-7.36 (m, 5H, C ₆ H ₅ ), 6.126 and 5.902 (2xs, 1H, benzylidene-CH), 5.066 and 4.987 (2xs, 1H, sugar-H1), 4.396 (dd, 0.5H, J = 6.4 , 5.6, Sugar-H3), 4.248 (dd, 0.5H, J = 6.4, 6.0, Sugar-H3), 4.214 (dd, 0.5H, J = 6.0, Sugar-H2), 4.102 (dd, 0.5H, J = 4.8, sugar -H2), 3.86-3.38 (m, 4H ), 3.466 and _{3.424 (2xs, 3H, CH 3} 0), 2.984 (br s, 1H, OH). 13 C NMR (CDCl 3, 100 MHz) : 138.01, 136.41, 129.38, 128.99, 128.24, 128.12, 126.41, 125.86, 103.85, 102.60, 97.74, 97.53, 79.36, 77.75, 77.47, 74.81, 70.00, 68.91, 68.42, 66.97, 54.78, 51.18 and 51.10.

Step b: Methyl 6-azido-3-O-benzyl-6-deoxy-α-D-mannopyranoside and methyl 6-azido-2-O-benzyl-6-deoxy-α-D-mannopyranoside methyl 6 -azido-6 A solution of DMF (7.8 ml, 0.1 M) containing 2-deoxy-2,3-di-O-benzylidene-α-D-mannopyranoside (240 mg, 0.781 mmol) was added to sodium cyanoborohydride (589 mg, 9.37 mmol, 12 Equivalent) and 3Å molecular sieves (1 g). The mixture was stirred at room temperature for 20 minutes and at 70 ° C. TFA (0.361 ml, 4.686 mmol, 6 eq) was added slowly. After the addition was complete, the mixture was stirred at 70 ° C. for 6 hours, cooled to 0 ° C. and then made basic by adding solid sodium carbonate. The cooled mixture was filtered and the resulting mass was washed with ethyl acetate. The filtered and washed product was extracted once with saturated sodium carbonate. The extract was purified by column chromatography (2.5 × 10 cm silica, eluting with 1: 4 to 1: 1 hexane: ethyl acetate) and concentrated to give gummy methyl 6-azido-3. -O-benzyl-6-deoxy-α-D-mannopyranoside (clear gum, 64 mg, 26%, Rf = 0.45, 1: 1 ethyl acetate: hexane), ie ¹ H NMR (CDCl ₃ , 400 MHz) : 7.41-7.32 (m, 5H, Ph), 4.78 (d, 1H, J _1-2 = 1.6, H1), 4.70 (d, 1H, J _gem = 12.0, CH ₂ ), 4.56 (d, 1H, CH ₂ ), 4.02 (dd, 1H, J _2-3 = 3.2, H2), 3.78 (dd, 1H, J _3-4 = 8.4, J _4-5 = 10.0, H4), 3.73 (ddd, 1H, J _{5 -6ax} = 6.0, J _5-6eq = 3.2, H5), 3.64 (dd, 1H, H3), 3.52 (dd, 1H, J _6ax-6eq = 13.2, H6eq), 3.47 (dd, 1H, J _6ax-6eq = 13.2, J _5-6ax = 6.0, H6ax), 3.40 (s, 3H, MeO), 2.21 (br s, 2H, 2xOH) and methyl 6-azido-2-O-benzyl-6-deoxy-α-D -Mannopyranoside (transparent waxy solid, 62 mg, 26%, Rf = 0.27, 1: 1 ethyl acetate Hexane), _{^{i.e., 1 H NMR (CDCl 3,}} 400 MHz): 7.38-7.28 (m, 5H, Ph), 4.78 (s, 1H, H1), 4.71 (d, 1H, J = 11.6, CH 2), 4.53 (d, 1H, J = 11.6, CH ₂ ), 3.75-3.61 (m, 4H, H2, H3, H4 and H5), 3.53 (dd, 1H, J = 13.2, 1.5, H6eq), 3.46-3.40 ( dm, 1H, J = 13.2, H6ax), 3.38 (s, 3H, MeO), 2.83 (br s, 2H, 2xOH).

Step c: Methyl 6-azido-2-O-benzyl-6-deoxy-2,3-di-O-sulfonate-α-D-mannopyranoside, disodium salt (PG2160)
Methyl 6-azido-2-O-benzyl-6-deoxy-α-D-mannopyranoside was sulfonated by the general procedure for sulfonation to give 59%, 64 mg of the title compound as a white powder. It was. ¹ H NMR (D ₂ O, 400 MHz): 7.38-7.26 (m, 5H, Ph), 4.72 (d, 1H, J _gem = 12.0, CH ₂ Ph), 4.59 (d, 1H, J _1-2 = 2.0, H1), 4.58 (d, 1H, CH ₂ Ph), 4.49 (d, 1H, J _2-3 = 2.8, J _3-4 = 9.6, H3), 4.44 (dd, 1H, J _4-5 = 9.6, H4), 4.07 (dd, 1H, H2), 3.78 (ddd, J _5-6ax = 5.6, J _5-6eq = 2.4, H5), 3.62 (dd, 1H, J _6ax-6eq = 13.2, H6eq) , 3.51 (dd, 1H, J _6ax-6eq = 13.2, J _5-6ax = 5.6, H6ax), 3.22 (s, 3H, MeO).

Step d: Methyl 6-azido-3-O-benzyl-6-deoxy-2,4-di-O-sulfonate-α-D-mannopyranoside, disodium salt (PG2161)
Methyl 6-azido-3-O-benzyl-6-deoxy-α-D-mannopyranoside (64 mg) was sulfonated by the general procedure for sulfonation to give 66%, 58 mg of the title compound as a white powder was gotten. ¹ H NMR (D ₂ O, 400 MHz): 7.42-7.21 (m, 5H, Ph), 4.92 (d, 1H, J _1-2 = 2.4, H1), 4.67 (d, 1H, J _gem = 12.4, CH ₂ Ph), 4.60 (d, 1H, CH ₂ Ph), 4.56 (dd, 1H, J _2-3 = 3.2, H2), 4.35 (dd, J _3-4 = 9.6, J _4-5 = 9.6, H4), 3.80 (dd, 1H, H3), 3.75 (ddd, J _5-6ax = 6.0, J _5-6eq = 2.8, H5), 3.66 (dd, 1H, J _6ax-6eq = 13.4, H6eq), 3.54 (dd, 1H, J _6ax-6eq = 13.4, J _5-6ax = 6.0, H6ax), 3.29 (s, 3H, MeO).

Step e: Methyl 6- [1 ′-(4-phenyl) triazolyl] -2-O-benzyl-6-deoxy-2,3-di-O-sulfonate-α-D-mannopyranoside, disodium salt (PG2173)
When methyl 6-azido-2-O-benzyl-6-deoxy-2,3-di-O-sulfonate-α-D-mannopyranoside, disodium salt was subjected to the reaction procedure of Wisgen using phenylacetylene, 67% gave 6.8 mg of the title compound as a white powder, Rf of 0.34 with ethyl acetate-methanol-water = 10: 2: 1. ¹ H NMR (D ₂ O, 400 MHz): 8.26 (s, 1H, triazole), 7.64-7.60 (m, 2H), 7.37-7.15 (m, 8H), 4.89 (dd, 1H, J = 14.4, 2.8 , H3), 4.67 (d, 1H, J = 12.0, PhCH ₂ ), 4.57-4.39 (m, 5H, PhCH ₂ , H 1, H4 and H6eq and H6ax), 4.04 (dd, 1H, J = 2.8, 2.0 , H2), 3.99 (ddd, 1H, J = 9.2, 8.8, 2.4, H5), 2.83 (s, 3H, MeO).

Example 19: PG2170
Aryl 6-azido-2,3-O-disulfonate-6-deoxy-4-O- (1-naphthylmethyl) -α-D-mannopyranoside, disodium salt (containing 10% of 2-naphthylmethyl isomer) ) (PG2170)
Analogous to the procedure for the preparation of PG2163 from aryl 6-azido-6-deoxy-α-D-mannopyranoside, 87%, 87.5 mg of the title compound are obtained, ethyl acetate-methanol-water = 10: The Rf at 2: 1 was 0.28 (major) and 0.22 (minor). ¹ H NMR (D ₂ O, 400 MHz): 7.87 (d, 1H, J = 8.4, naphthyl), 7.65-7.54 (m, 2H, naphthyl), 7.33-7.19 (m, 4H, naphthyl), 5.68 (ddt , J aryl _2'-3'trans = 22.0, J aryl _2'-3'cis = 10.8, J aryl _{1'-2 '} = 6.0, aryl-2'), 5.23 (AB quadruple, 2H, J _gem = 12.0, naphthyl-CH ₂ ), 5.17-5.02 (m, 3H, aryl-3 'and H1), 4.74 (dd, 1H, J _1-2 = 2.0, J _2-3 = 3.2, H2), 4.64 ( d, 1H, J _3-4 = 9.6, H3), 3.90 (dd, 1H, J aryl _1'gem = 13.2, J = 6.0, aryl 1 '), 3.81 (dd, 1H, aryl-1'), 3.64 (t, 1H, J _4-5 = 9.6, H4), 3.39 (ddd, 1H, J _5-6eq = 2.0, J _5-6ax = 5.2, H5), 2.78 (dd, 1H, J _6eq-6ax = 13.6 , H6eq), 2.68 (dd, 1H, J _5-6ax = 5.2, J _6eq-6ax = 13.6, H6ax). The typical signal with the 2-naphthylmethyl derivative, one of the isomers, is 4.84 (AB Quadruple, 2H, J _gem = 11.2, naphthyl-CH ₂ ), 3.52 (ddd, J _4-5 = 10.0, J _5-6eq = 2.4, J _5-6ax = 6.0, H5), 3.07 (dd, 1H _J6eq-6ax = 13.6, H6eq), 2.96 (dd, 1H, _J6eq-6ax = 13.6, _{J5 -ax} = 6.0, H6ax). This isomer is commercially available as a 9: 1 mixture of 1- and 2-bromomethylnaphthalene used in step a.

  Other compounds have also been synthesized using synthetic methods with suitable modifications to the procedures detailed in Examples 1-19 above. The following tables showing the results of biological tests of the compounds of the present invention are exemplified.

Example 20
Compound biological testing
Method
1. Using a surface plasmon resonance apparatus (SPR) that utilizes the affinity of the growth factor binding solution, the binding of ligands involved in the growth factor was analyzed. In the principle of this analysis method, heparin immobilized on the surface of the sensing element utilizes the action of discriminating between a free growth factor and a binding growth factor in an equilibration solution containing a growth factor and a ligand. When this solution is injected and free growth factor binds to immobilized heparin, the SPR response value increases and the concentration is determined. The decrease in the concentration of free growth factor brought about by the effect of the ligand concentration makes it possible to calculate the dissociation constant, K _d value. It is important that the binding of the ligand to the growth factor is observed only when the heparin binding site is involved in the interactivity, and therefore, non-specific binding to other sites of the protein. The need for analysis is eliminated. All protein: ligand interactions are believed to have a 1: 1 stoichiometric relationship.

A method for preparing a heparin-coated sensing element using immobilization of biotinylated BSA-heparin to a sensing element coated with streptavidin has been disclosed [24]. Heparin can be immobilized using aldehyde linkages using either adipic acid dihydrazide or 1,4-diaminobutane. To determine each K _d value, a solution was prepared from a buffer containing a specific concentration of protein and various concentrations of ligand. Ligand binding to FGF-1 and VEGF was achieved with HBS-EP buffer (10 mM HEPES, pH 7.4, 150 mM sodium chloride, 3.0 mM EDTA and 0.005% (v / v) polysorbate 20), and FGF-2 Ligand binding was analyzed with HBS-EP buffer containing 0.3 M sodium chloride [24]. Prior to injection, samples were stored at 4 ° C. to maximize protein stability. In the analysis of each mixture, 50-200 μl of solution was injected at a rate of 5-40 μl / min and relative binding response values were measured. All surface binding tests were performed at 25 ° C. The surface was regenerated by injecting 40 μl of 4M sodium chloride at a rate of 40 μl / min and then injecting 40 μl of buffer at a rate of 40 μl / min.

  Sensorgram data was analyzed using BIAevaluation software (BIAcore).

  To obtain a curve for specific binding, the sensorgram initial value was subtracted from the sensorgram experimental value, and the baseline was corrected sequentially from zero to all curves. The relative binding response value for each infusion test is given by the following formula:

式中、γは、相対的結合応答値であり、また、γ_mは、最大の結合応答値である数式を用いて、遊離タンパク質濃度から換算した。 注入前の溶液内に存在する結合平衡状態は、１：１の化学量論的関係にあると考えられる。 従って、

In the formula, γ is a relative binding response value, and γ _m is converted from the free protein concentration using a mathematical formula which is the maximum binding response value. The binding equilibrium existing in the solution before injection is considered to be in a 1: 1 stoichiometric relationship. Therefore,

式中、Ｐは成長因子タンパク質、Ｌはリガンド、Ｐ・Ｌはタンパク質：リガンド錯体である平衡状態での平衡式は、

In the formula, P is a growth factor protein, L is a ligand, and P · L is a protein: ligand complex.

であり、結合方程式[２４]は、

And the coupling equation [24] is

として表すことができる。

Can be expressed as

The K _d value shown here is a numerical value derived from the curve drawn by the [P] value and the [L] total amount value value using a coupling equation. When the _Kd value is acquired twice, the numerical value shown here uses the average value of the actually measured values. GAG mimetics that bind tightly to these growth factors are known to elicit biological responses in vivo [24].

2. Antiviral analysis According to the method described in Nyberg et al. [25] regarding the prevention of viral infection and the transmission of viruses between cells, against two types of herpes simplex virus (HSV), namely HSV-1 and HSV-2 The selected compounds were tested. A monolayer culture of African green monkey kidney cells (GMK AH1) [26] cultured in 6-well cluster plates was used. The virus strains used here were type 1 herpes simplex virus (HSV-1) KOS321 strain [27] and HSV-2 strain 333 [28].

  In both analyses, 200 μm compounds were subjected to testing.

  (i) For analysis of HSV infection, these compounds coexist with the virus, incubated at room temperature for 10 minutes, and these mixtures are added to the cells to allow (block) viral attachment / invasion to the cells. For just that purpose, it was left as it was for 1 hour. In other words, this analysis reveals the effect of the compounds subjected to the test, ie, the presence or absence of the effect of binding to the virus particles and the effect of inhibiting the attachment / entry of the virus to the cells. The inhibitory effect can be confirmed by the decrease in the number of virus plaques.

  (ii) The next analysis, also referred to as HSV transmission, is a method of adding compounds to cells after virus attachment / invasion has been tried. In addition to the effect of the compounds used in the test, ie, the effect of inhibiting the propagation from infected cells to uninfected cells (cell-to-cell propagation), this analysis is the effect of invading cells and inhibiting virus replication. Clarify the presence or absence of The case where the virus infection analysis shows no effect and the virus transmission analysis shows the activity suggests that the compound has entered the cell and inhibited the virus replication process. The inhibitory effect can be confirmed by reducing the size of the virus plaque.

  The test results (see Table 5) show the percentage (%) relative to the control, ie the number of virus plaques formed in the presence of the compound (infection analysis) relative to the sham-treated control (no compound used). ) Or size (propagation analysis).

Results The results of the tests described in the previous section are summarized in Tables 1 to 5 below.

表１〜表４に記載の結果は、本願発明が提供する多種多様な化合物が、ＧＡＧ結合性成長因子に対して顕著な親和性を示し、また、それらの活性に関するモジュレーターとしても機能することを実証するものである。 また、表５に記載の結果は、これら化合物が、in vivo活性を実際に有していることを実証している。

The results described in Tables 1 to 4 show that a wide variety of compounds provided by the present invention show significant affinity for GAG-binding growth factors and also function as modulators of their activity. It is a demonstration. The results listed in Table 5 also demonstrate that these compounds actually have in vivo activity.

  The above-described embodiments are merely for the purpose of exemplifying the outline of the present invention, and those skilled in the art can easily conceive various modifications and changes. The present invention can be implemented and reproduced in various ways, and can also be implemented and reproduced according to embodiments other than those described above. The terminology used in the present specification is only for the purpose of describing the present invention, and the present invention should not be construed as being limited thereto.

  As used herein, the term “comprising” and its synonyms such as “including” or “including” refer to the state of incorporating the element or elements referred to therein. Unless there is a specific interpretation about the term, it is not intended to exclude other elements or elements.

  Although the documents cited in the present specification are listed, this does not confirm that the present invention is configured based on common general technical knowledge in Australia.

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Claims (14)

The following formula:

Wherein each independent X is CH ₂ , C (O), N, O, S, S (O), S (O) ₂ or a chemical bond, and independent R _{1 to} R ₅ each chemical bond, or hydrogen; halogen; azide; C ₁ -C ₈ alkyl or alkenyl, aryl or heteroaryl, and optionally, alkoxy, aryl, heteroaryl or aryloxy group; -COOH, -S R group further substituted by (O) ₂ OH, phosphate, carboxylate or tetrazolyl; -S (O) ₂ OH, -S (O) OH, -S (O) R, -S (O) ₂ R, —S (O) ₂ NH ₂ , —S (O) ₂ OR, —S (O) OR; —C (O) R; phosphate, carboxylate or tetrazolyl; unsubstituted or substituted heterocyclic compounds, i.e., an alkyl or aryl group, and, -CH ₂ NHC (O) R with R group described above, -C _{2 N (C (O) R} ) 2, -CH 2 OR; binding to different R ₁ to R ₅ to form a new cyclic group; the following formulas, namely;

Wherein Y is hydrogen and R or —C (O) R having the aforementioned R group; at least one and more than two Not independent R _{7 to} R ₁₁ have the structure described in Formula I; or form a chemical structure comprising the second unit described in Formula II via the “Y” group;
Further, each independent unit is such that when R ₁ is —CH ₃ , —S (O) ₂ OH or —H, at least one of R _{2 to} R ₅ is —H or —S (O) ₂ OH. And when no structure of Formula II exists and any of R _{1 to} R ₅ does not form an anhydrous bridge, two or less of R _{1 to} R ₅ are —S (O) ₂ OH And a chemical structure having the requirement that the stereochemistry of Formula I is substituted by R _{7 to} R ₁₀ under the condition that it is not gluco or galacto,
A compound selected from the group consisting of:   The compound according to claim 1, which is PG2024, PG2037, PG2173, PG2198 described in the present specification.   The compound according to claim 1, which is any one of the compounds described in Tables 1 to 4 of the present specification.   A pharmaceutical or veterinary composition for the prevention or treatment of diseases caused by coagulation, thrombosis and / or microbial infection in a mammal, together with at least one compound as claimed in claim 1, A composition comprising a pharmaceutically or veterinary acceptable carrier or diluent for at least one of said compounds.   5. The composition of claim 4, further comprising a pharmaceutically or veterinarily acceptable excipient, buffer, stabilizer, isotonic agent, preservative or antioxidant.   5. A composition according to claim 4, containing the compound as an ester, free acid or free base, hydrate or prodrug.   Use of a compound according to claim 1 for the manufacture of a medicament for the prevention or treatment of diseases caused by angiogenesis, tumor metastasis, microbial infection in mammals.   8. Use according to claim 7, wherein the mammal is a human.   A method for preventing or treating a disease caused by angiogenesis, tumor metastasis, inflammation, coagulation, thrombosis and / or microbial infection in a mammal, comprising at least one compound according to claim 1 or Administering to the mammal an effective amount of a composition comprising at least one said compound.   The method according to claim 9, wherein the mammal is a human.   10. The method of claim 9, wherein the disease resulting from angiogenesis is angiogenesis resulting from proliferative retinopathy or solid tumor growth.   10. The method of claim 9, wherein the disease resulting from inflammation is due to rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, graft rejection or chronic asthma.   10. The method of claim 9, wherein the disease resulting from clotting and / or thrombosis is due to deep vein thrombosis, pulmonary embolism, thrombotic stroke, peripheral arterial thrombosis, unstable angina or myocardial infarction. The method according to claim 9, wherein the disease caused by viral infection is herpes simplex.