Use of heparin trisaccharide compound in preparation of drug for treating chronic obstructive pulmonary disease

Abstract

A trisaccharide compound having a core structure of Glc(1→4)IdoA(1→4)GlcNS and a use thereof in the preparation of a drug for treating COPD. The trisaccharide compound has almost no anticoagulant activity, does not have platelet factor 4 binding activity, and exhibits an extremely low bleeding risk during treatment.

Description

Application of heparin trisaccharide compounds in the preparation of drugs for treating chronic obstructive pulmonary disease

[0001] This application claims priority to an earlier application filed on December 31, 2024, with patent application number 202411975922.4, entitled "Use of a heparin trisaccharide compound in the preparation of a medicament for treating chronic obstructive pulmonary disease". The entire contents of the earlier application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of biomedical technology, specifically relating to the application of trisaccharide compounds in the preparation of drugs for the treatment or prevention of chronic obstructive pulmonary disease. Background Technology

[0003] Chronic obstructive pulmonary disease (COPD) is a disease characterized by airflow limitation, which is usually progressive and associated with abnormalities in the airways and alveoli. It is primarily caused by long-term inhalation of harmful particles or gases, most commonly cigarette smoke, indoor or outdoor air pollution, and chronic bronchitis. The history of COPD dates back over 200 years. In 1679, Bonet first described COPD as "a large lung." In 1789, Baillie published a series of pathological illustrations of the lungs in emphysema. In 1821, Emphysema is described, and the combination of emphysema and chronic bronchitis is mentioned. COPD is a leading cause of death worldwide, often referred to as the "silent killer" due to its insidious onset. Treatment of COPD faces numerous challenges and dilemmas. Drug therapy primarily relies on bronchodilators, which can improve symptoms and quality of life and prevent acute exacerbations to some extent. However, anti-inflammatory treatment targeting the underlying cause of the disease remains unresolved, especially given the less-than-ideal efficacy of inhaled corticosteroids, necessitating the development of new drugs and treatment methods. Summary of the Invention

[0004] This invention provides a trisaccharide compound. Experimental data from this invention demonstrate that, in a COPD disease model, this compound can improve the decline in lung function in COPD patients by reducing chronic inflammation and airway remodeling, and is expected to become a new drug for the treatment of COPD.

[0005] The trisaccharide compound of formula A or its salt, with the three monosaccharides from left to right represented by B, C, and D respectively:

[0006] in:

[0007] R1, R2, R3, and R4 may be the same or different, and are independently selected from H or -SO3H.

[0008] R5 is a -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H.

[0009] R6 is -OH, -OSO3H, or -NHSO3H.

[0010] The salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation.

[0011] The monovalent cation is, for example, selected from Na. + K + Li + NH4 + etc.; the divalent cation is selected, for example, from Ca... 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ etc.; the trivalent cations are selected, for example, from Al 3+ Fe 3+ Etc. Those skilled in the art will understand that when the compound forms a salt, the corresponding group in formula A is its anionic group, such as -COO. - -SO3 - .

[0012] The compound of formula A is a compound with a single optical activity, namely Glc(1→4)IdoA(1→4)GlcNS, where the D sugar terminal group is α or β configuration.

[0013] In some embodiments of the present invention, in formula A, R1 is selected from H or -SO3H; R2, R3 and R4 are the same, all of which are H; R6 is -OH, -OSO3H or -NHSO3H.

[0014] In some embodiments of the present invention, in formula A, R1, R2, R3 and R4 are the same, all being H; R6 is -NHSO3H.

[0015] In some other embodiments of the present invention, in formula A, R1 is -SO3H; R2, R3 and R4 are the same, all of which are H; R6 is -OH.

[0016] In some embodiments of the present invention, in formula A, R5 is -C1-3 alkyl, -C2-4 alkylene-NH2, or -C2-4 alkylene-NHSO3H. Preferably, R5 is methyl, ethyl, -C2H4NH2, -C2H4NHSO3H, -C3H6NH2, or -C3H6NHSO3H.

[0017] In some embodiments of the present invention, the salt is a salt formed with a monovalent cation, wherein the monovalent cation is selected from Na. + K + Li + NH4 + Preferably, the monovalent cation is selected from Na. + K + .

[0018] In some embodiments of the present invention, the salt is a salt formed with a divalent cation selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ Preferably, the divalent cation is Ca. 2+ .

[0019] In some embodiments of the present invention, in formula A, R1, R2, R3, and R4 are the same, all being H; R5 is -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H; R6 is -NHSO3H, and the salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation. Preferably, R5 is -C1-3 alkyl, -C2-4 alkylene-NH2, or -C2-4 alkylene-NHSO3H; more preferably, R5 is methyl, ethyl, -C2H4NH2, -C2H4NHSO3H, -C3H6NH2, or -C3H6NHSO3H. Preferably, the salt is a salt formed with a monovalent cation selected from Na. + K + Li + NH4 + Preferably, the monovalent cation is selected from Na. + K + Alternatively, the salt is a salt formed with a divalent cation selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ Preferably, the divalent cation is Ca. 2+ .

[0020] In some embodiments of the present invention, in formula A, R1 is -SO3H; R2, R3, and R4 are the same, all being H; R6 is -OH; R5 is -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H; the salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation. Preferably, R5 is -C1-3 alkyl, -C2-4 alkylene-NH2, or -C2-4 alkylene-NHSO3H; more preferably, R5 is methyl, ethyl, -C2H4NH2, -C2H4NHSO3H, -C3H6NH2, or -C3H6NHSO3H. Preferably, the salt is a salt formed with a monovalent cation selected from Na. + K + Li + NH4 + Preferably, the monovalent cation is selected from Na. + K + Alternatively, the salt is a salt formed with a divalent cation selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ Preferably, the divalent cation is Ca. 2+ .

[0021] In one embodiment of the present invention, the structural formula of compound A is as follows, referred to as CV010 in the present invention.

[0022] In one embodiment of the present invention, the sodium salt structure of compound A is as follows, referred to as CV016 in the present invention.

[0023] In another embodiment of the present invention, the structural formula of compound A is as follows, and is referred to as TSC in the present invention.

[0024] In another embodiment of the invention, the sodium salt of compound A has the following structural formula, referred to as TSC19 in this invention.

[0025] According to the present invention, the compound of formula A is derived via a fully protected trisaccharide intermediate of formula E. [E] is obtained by sequentially undergoing dehydroxyl protecting group removal, O-sulfonation, optional azide reduction reaction, and finally N-sulfonation. Those skilled in the art will understand that after obtaining compound A, the corresponding salt of the compound can be obtained by salting it with a cation, for example, by exchanging compound A with a cation exchange resin to prepare its corresponding monovalent, divalent, or trivalent salt.

[0026] In the fully protected trisaccharide intermediate E, Rx can be an azide group, an -O-acyl group (including but not limited to chloroacetyl, acetyl, benzoyl, pivaloyl, etc.), or an -O-benzyl group. 11 R 21 R 31 R 41 R 51 and R 61 They can be the same or different, and are independently selected from chloroacetyl, acetyl, benzoyl, pivaloyl, benzyl, and p-methoxybenzyl. In one embodiment, Rx is preferably an azide group, R 11 R 21 R 31 and R 51 Same, R 41 and R 61 Same or different, and both are different from R 11 R 21 R 31 and R 51 Further optimization preferred Rx to be an azide group, R 11 R 21 R 31 and R 51 They are the same, both are benzyl, R 41 and R 61 The groups, whether the groups are the same or different, are independently selected from acetyl and benzoyl groups. In another embodiment, Rx is preferably -O-benzyl, and R... 21 R 31 and R 51 Same, R 11 R 41 and R 61 Same or different; further preferred Rx is -O-benzyl, R 21 R 31 and R 51 They are the same, both are benzyl, R 11 R 41 and R 61 Whether the groups are the same or different, they are independently selected from acetyl and benzoyl groups.

[0027] Preferably, in the reaction, all hydroxyl groups to be sulfonated are deprotected in one step, and after the hydroxyl O-sulfonation, all hydroxyl groups not to be sulfonated are deprotected in one step.

[0028] In a specific embodiment of the present invention, the present invention uses the synthesis method described above to synthesize CV010 and CV016: Rx, R 11 R 21 R 31 R 41 R 51 and R 61 The definitions are the same as before, and the reaction pathway is as follows:

[0029] In one specific embodiment of the preparation of CV010 and CV016, R 11 R 21 R 31 and R 51 All are benzyl, R 41 For acetyl, R 61 The radical is benzoyl, and Y is selected from H and Na. + K + Li + NH4 + H or Na are preferred. + .

[0030] In another specific embodiment of the present invention, the present invention uses the aforementioned synthesis method to synthesize TSC and TSC19:Rx, R 11 R 21 R 31 R 41 R 51 and R 61 The definitions are the same as before, and the reaction pathway is as follows:

[0031] In one specific embodiment of the preparation of TSC and TSC19, R 21 R 31 and R 51 All are benzyl, R 11 and R on D sugar 41 For acetyl, R 61 and R on β sugar 41 The radical is benzoyl, and Y is selected from H and Na. + K + Li + NH4 + H or Na are preferred. + .

[0032] The fully protected trisaccharide intermediate E can be derived from the monosaccharide intermediate F. [F] and disaccharide receptor 4 It is obtained through a glycosylation reaction.

[0033] Rx can be an azide group, an -O-acyl group (including but not limited to chloroacetyl, acetyl, benzoyl, pivaloyl, etc.), or an -O-benzyl group. 11 R 21 R 31 R 41 R 51 and R 61 They can be the same or different, and are independently selected from chloroacetyl, acetyl, benzoyl, pivaloyl, benzyl, and p-methoxybenzyl; X is a leaving group suitable for reacting with other acceptors to form bonds between glycosides.

[0034] Preferably, X is a hydroxyl group, a thioalkyl group, a thioaryl group, a halogen, a trichloroimine acetyl group, a phosphate ester, or a tert-butyldiphenylsilyloxy group.

[0035] Rx is preferably an azide group, R 11 R 21 R 31 and R 51 They are the same, both are benzyl, R 41 and R 61 Whether the Rx group is the same or different, it is independently selected from acetyl or benzoyl; or, preferably, Rx is -O-benzyl, R 21 R 31 and R 51 They are the same, both are benzyl, R 11 R 41 and R 61 Whether the groups are the same or different, they are independently selected from acetyl and benzoyl groups.

[0036] According to the present invention, the glycosylation reaction temperature is -80°C to -10°C. The reaction can be carried out under strong acid conditions, such as trifluoromethanesulfonic acid, TBSOTf, TMSOTf, etc.

[0037] In one embodiment of the present invention, the fully protected trisaccharide intermediate 1 is obtained by glycosylation of a monosaccharide intermediate 3 and a disaccharide acceptor 4, as shown in the following reaction formula:

[0038] In one embodiment of the present invention, the fully protected trisaccharide intermediate 2 is obtained by glycosylation of monosaccharide intermediate 2-1 and disaccharide acceptor 4, as shown in the following reaction formula:

[0039] The disaccharide intermediate 4 can be obtained from the monosaccharide intermediate 6 and the monosaccharide intermediate 7, as shown in the following reaction formula:

[0040] The monosaccharides 6, 7, and 4 disaccharide receptor used in the above preparation method can be prepared according to synthetic methods known in the art, such as: Preactivation-based, iterative one-pot synthesis of anticoagulant pentasaccharide fondaparinux Sodium. Org. Chem. Front., 2019, 6, 3116; Total Synthesis of Anticoagulant Pentasaccharide Fondaparinux. ChemMedChem, 2014, 9, 1071–1080. The definitions of the functional groups in monosaccharides 6, 7, and 4 are the same as those of the corresponding functional groups described above.

[0041] Therefore, the present invention also provides the intermediates in the above synthesis method and their preparation methods.

[0042] A monosaccharide intermediate 3 has the following structure: Among them, R is preferred. 11 and R 31 All are benzyl, R 41 It is chloroacetyl, acetyl, benzoyl, or pivaloyl. In one embodiment of the invention, the monosaccharide intermediate 3 has R... 11 and R 31 All are benzyl, R 41 It is an acetyl group, which can be either α or β configuration, named 3-1, and has the following structural formula:

[0043] The reaction formula for its preparation method is as follows:

[0044] A monosaccharide intermediate 2-1 has the following structure: Among them, R is preferred. 31 It is benzyl, R 11 and R 41 Independently selected from chloroacetyl, acetyl, benzoyl, or pivaloyl. In one embodiment of the invention, the monosaccharide intermediate 2-1, R... 31 It is benzyl, R 11 For acetyl, R 41 It is a benzoyl group, which can be either α-configuration or β-configuration, named F-1, with the structural formula as follows:

[0045] Its preparation method is as follows:

[0046] A disaccharide intermediate 4 has the following structure: Among them, R is preferred. 21 and R 51 It is benzyl, R 41 and R 61 The same or different, selected from chloroacetyl, acetyl, benzoyl, or pivaloyl. In one embodiment of the invention, the R of the disaccharide intermediate 4... 21 and R 51 It is benzyl, R 41 For acetyl, R 61 It is benzoyl, named 4-1, and has the following structural formula: 4-1.

[0047] Disaccharide intermediate 4-1 can be prepared using synthetic methods known in the art, such as Total Synthesis of Anticoagulant Pentasaccharide Fondaparinux. ChemMedChem, 2014, 9, 1071–1080.

[0048] A fully protected trisaccharide intermediate 1 has the following structural formula:

[0049] Where R 11 R 21 R 31 R 41 R 51 and R 61 They may be the same or different, and are independently selected from chloroacetyl, acetyl, benzoyl, pivaloyl, benzyl, and p-methoxybenzyl. In one specific embodiment of the invention, R 11 R 21 R 31 and R 51 All are benzyl, R 41 For acetyl, R 61 It is benzoyl.

[0050] The present invention also provides intermediates I, II and III.

[0051] The structure of compound I is as follows:

[0052] Among them, R 11 R 21 R 31 and R 51 They can be the same or different, and are independently selected from chloroacetyl, acetyl, benzoyl, pivaloyl, benzyl, and p-methoxybenzyl. Preferably, R 11 R 21 R 31 and R 51 All are benzyl groups.

[0053] The structure of compound II is as follows:

[0054] Among them, R 11 R 21 R 31 and R 51 They may be the same or different, and are independently selected from chloroacetyl, acetyl, benzoyl, pivaloyl, benzyl, and p-methoxybenzyl, with Y selected from H and Na. + K + Li + NH4 + R is preferred 11 R 21 R 31 and R 51 All are benzyl groups, and Y is H or Na. + .

[0055] The structure of compound III is as follows:

[0056] Where Y is selected from H and Na + K + Li + NH4 + Preferably, Y is H or Na. + .

[0057] Intermediates I, II, and III are generated from trisaccharide intermediate 1 through sequential dehydroxylation, sulfonation, and azide reduction, as shown in the following reaction formulas:

[0058] The dehydroxyl protecting group, sulfonation, and azide reduction can all be carried out using reaction methods and conditions known in the art. In one embodiment of the invention, intermediate 1, under alkaline conditions, simultaneously removes R. 41 R 61 Intermediate I is obtained by reacting methyl ester with SO3·NMe3. In one embodiment of the invention, intermediate I is subjected to SO3·NMe3 to obtain O-sulfonated intermediate II. In one embodiment of the invention, intermediate II undergoes catalytic hydrogenation to remove the benzyl and Cbz groups, while simultaneously reducing the azide to generate an amino group, yielding intermediate III. In one embodiment of the invention, the amino group in compound III is sulfonated under SO3·Py to obtain compound CV010, which is then ion-exchanged using a sodium-type ion exchange resin to obtain compound CV016. The sodium ion exchange resin can be a resin known in the art, including but not limited to Amberlite IR120 Na. + Dowex-50-WX4-Na +wait.

[0059] A fully protected trisaccharide intermediate 2 has the following structural formula:

[0060] Among them, R 11 R 21 R 31 and R 51 They may be the same or different, and are independently selected from chloroacetyl, acetyl, benzoyl, pivaloyl, benzyl, and p-methoxybenzyl. In one specific embodiment of the invention, R 21 R 31 and R 51 All are benzyl, R 11 and R on D sugar 41 For acetyl, R 61 and R on β sugar 41 It is benzoyl.

[0061] The intermediates 2-I, 2-II, and 2-III have the following structures, and are respectively generated from the trisaccharide intermediate 2 through sequential dehydroxylation, sulfonation, and reduction, as shown in the following reaction formulas, wherein the definitions of each group are as described above:

[0062] The dehydroxyl protecting group, sulfonation, and reduction can all be carried out using reaction methods and conditions known in the art. In one embodiment of the invention, intermediate 2, under alkaline conditions, simultaneously removes R. 11 R 41 R 61 Intermediate 2-I is obtained by reacting methyl ester with SO3·NMe3. In one embodiment of the invention, intermediate 2-I is subjected to SO3·NMe3 to obtain O-sulfonated intermediate 2-II. In one embodiment of the invention, intermediate 2-II is subjected to catalytic hydrogenation to remove the benzyl group and Cbz, yielding intermediate 2-III. In one embodiment of the invention, the amino group in compound 2-III is sulfonated under the action of SO3·Py to obtain compound TSC, which is then ion-exchanged with a sodium-type ion exchange resin to obtain compound TSC19. The sodium ion exchange resin may be a resin known in the art, including but not limited to Amberlite IR120 Na. + Dowex-50-WX4-Na + wait.

[0063] COPD is a heterogeneous lung condition characterized by the presence of chronic and poorly reversible persistent (often progressive) airflow obstruction associated with an abnormal inflammatory response. Its main clinical symptoms are cough, sputum production, and shortness of breath or difficulty breathing when walking or increasing activity; these symptoms tend to worsen over the years.

[0064] According to the GOLD criteria (Global Initiative for Chronic Obstructive Lung Disease), a forced expiratory volume in one second (FEV1) to forced vital capacity (FVC) ratio < 0.7 after inhaling a bronchodilator indicates persistent airflow limitation and can be diagnosed as COPD. The severity of airflow limitation is assessed based on the percentage of predicted FEV1, categorized into four levels: Mild (GOLD 1): FEV1 ≥ 80% of predicted value. Patients at this stage are usually asymptomatic or experience only mild shortness of breath during strenuous exercise; Moderate (GOLD 2): 50% ≤ FEV1 < 80% of predicted value. Patients experience progressively worsening shortness of breath, especially during daily activities; Severe (GOLD 3): 30% ≤ FEV1 < 50% of predicted value. Patients experience exacerbated shortness of breath, significant limitation of daily activities, and may experience acute exacerbations; Very Severe (GOLD 4): FEV1 < 30% of predicted value, or accompanied by symptoms of chronic respiratory failure. Patients at this stage experience a severe decline in quality of life and may require long-term oxygen therapy.

[0065] COPD can be classified according to its etiology as follows: genetically determined COPD (e.g., α1 antitrypsin deficiency), COPD caused by lung developmental abnormalities, environmental COPD (including smoking-related COPD, COPD related to exposure to biological dyes and pollution), and infection-related COPD (e.g., tuberculosis-related COPD, childhood infection, COPD with chronic bronchitis, etc.).

[0066] The compound of formula A of the present invention, or its salt or solvate thereof, can inhibit the inflammatory response in COPD, reduce the pathological changes in lung tissue of COPD (reduce inflammatory cell infiltration around the bronchi and blood vessels, reduce the shedding of alveolar epithelial cells, reduce bronchial mucosal edema, and protect alveolar tissue structure), reduce collagen deposition in the airways, and improve airflow obstruction.

[0067] This invention provides the use of a compound of formula A, or a salt thereof, or a solvate thereof, in the preparation of a medicament for the treatment or prevention of COPD.

[0068] The present invention also provides a compound of formula A or a salt thereof or a solvate thereof for the treatment or prevention of COPD.

[0069] The present invention also provides a compound of formula A or a salt thereof or a solvate thereof for the treatment or prevention of COPD.

[0070] In some embodiments of the present invention, the COPD is mild CPOD, moderate COPD, or severe COPD.

[0071] In some embodiments of the present invention, the COPD is environmental COPD, caused by infection.

[0072] In some embodiments of the present invention, treatment with a compound of formula A or a salt thereof or a solvate thereof improves and / or stabilizes at least one of the respiratory clinical symptoms in COPD patients, including but not limited to cough, sputum production, dyspnea or shortness of breath when walking or increasing activity.

[0073] The present invention also provides a pharmaceutical composition for treating or preventing COPD, comprising a compound of formula A of the present invention, or a salt thereof, or a solvate thereof, as an active ingredient, optionally further comprising one or more pharmaceutically acceptable carriers. The pharmaceutically acceptable carriers are various excipients commonly used or known in the pharmaceutical field, including but not limited to: diluents, binders, antioxidants, pH adjusters, preservatives, lubricants, disintegrants, etc.

[0074] The pharmaceutical composition may be administered in combination with other therapeutic agents or supportive therapies, or formulated as a combination drug, or may further contain other therapeutic agents. These other therapeutic agents are other medications for treating COPD.

[0075] The present invention also provides the use of a compound of formula A or a salt thereof or a solvate thereof in combination with other drugs for treating COPD in the preparation of a drug for treating or preventing COPD.

[0076] The present invention also provides the use of a compound of formula A or a salt thereof or a solvate thereof in the preparation of a medicament for the treatment or prevention of COPD in combination with other medicaments for the treatment of COPD.

[0077] The present invention also provides a method for treating or preventing COPD, characterized in that a therapeutically effective amount of a compound of formula A or a salt thereof or a solvate thereof, or a pharmaceutical composition containing a compound of formula A or a salt thereof or a solvate thereof, is administered to a patient in need.

[0078] The present invention also provides the use of a compound of formula A or a salt thereof or a solvate thereof, or the use of a pharmaceutical composition containing a compound of formula A or a salt thereof or a solvate thereof, for the treatment or prevention of COPD.

[0079] The present invention also provides the use of a compound of formula A or a salt thereof or a solvate thereof, or the use of a pharmaceutical composition containing a compound of formula A or a salt thereof or a solvate thereof, when treating or preventing COPD.

[0080] Other medications for treating COPD include, but are not limited to: glucocorticoids (such as budesonide), M receptor blockers (such as glycopyrronium bromide), and β receptor agonists (such as formoterol).

[0081] According to the present invention, the amount of compound A or its salt or solvate thereof in the pharmaceutical composition (calculated as compound A) is 0.1-1000 mg, preferably 1-500 mg, and more preferably 5-100 mg.

[0082] The mass percentage of compound A or its salt or solvate (calculated as compound A) in the pharmaceutical composition is 0.01%-95%, and may be 0.1%-10%, 0.3%-5%, or 10%-90% depending on the dosage form, preferably 20%-80%, more preferably 30%-70%, etc.

[0083] The dosage form of the pharmaceutical composition may be an oral dosage form, such as tablets, capsules, pills, powders, granules, suspensions, syrups, etc.; or an injectable dosage form, such as an injection solution, powder for injection, etc., administered via intravenous, intraperitoneal, subcutaneous, or intramuscular routes. All dosage forms used are well known to those skilled in the art of pharmaceutical science. For example, the pharmaceutical composition may be an injection solution, and the concentration of compound A or its salt or solvation in the injection solution, calculated based on compound A, may be 1-15 mg / ml, such as 5 mg / ml, 10 mg / ml, 12.5 mg / ml, etc.

[0084] The routes of administration of the pharmaceutical composition include, but are not limited to: oral; sublingual; sublingual; transdermal; pulmonary; rectal; parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, and intravenous; and by implantation of a reservoir or infusion device.

[0085] According to the present invention, the dosage of compound A or its salt or solvate (calculated as compound A) will depend on the recipient's age, health and weight, the type of concurrent drugs, the frequency of treatment, the route of administration, etc. The drug may be administered in a single daily dose, once daily, once every two days, once every three days, once every four days, or the total daily dose may be administered in two, three, or four separate doses per day. The dosage of compound A or its salt or solvate (calculated as compound A) is 0.01-100 mg / kg / day, preferably 0.1-10 mg / kg / day, for example 0.5 mg / kg / day, 1 mg / kg / day, 2 mg / kg / day, 5 mg / kg / day, etc.

[0086] Compound A of this invention exhibits a well-defined and highly effective therapeutic effect on COPD, but possesses almost no anticoagulant activity, high stability, and no platelet factor 4 binding activity. Therefore, it carries an extremely low risk of bleeding while being used for therapeutic purposes. The well-defined structure of compound A facilitates its preparation and quality control. Attached Figure Description

[0087] Figure 1. Analysis of inflammatory factor content in bronchoalveolar lavage fluid of the control group, model group (COPD), and treatment group (CV016 / TSC19) in the COPD model.

[0088] Figure 2 shows the therapeutic effect of CV016 / TSC19 on lung tissue in a COPD model, with representative HE-stained images of lung tissue from the control group, model group, and treatment group, respectively.

[0089] Figure 3 shows the therapeutic effect of CV016 / TSC19 on lung tissue in a COPD model, with representative Masson staining images of lung tissue from the control group, model group, and treatment group, respectively. Detailed Implementation

[0090] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0091] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0092] Ac: Acetyl; AgOTf: Silver trifluoromethanesulfonate; Bn: Benzyl; Bz: Benzoyl; ClAc: Monochloroacetyl; CSA: Camphorsulfonic acid; Cbz: Benzyloxycarbonyl; CsF: Cesium fluoride; DBU: 1,8-Diazabicycloundec-7-ene; DCM: Dichloromethane; DDQ: 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone; DMF: N,N-Dimethylformamide; PMB: p-Methoxybenzyl; TfOH: Trifluoromethanesulfonic acid; TBSOTf: Tert-butyldimethylsilyltrifluoromethanesulfonate; TEMPO: 2,2,6,6-Tetramethylpiperidine oxide; TMSOTf: Trimethylsilyltrifluoromethanesulfonate; Tol: Toluene.

[0093] Example 1: Preparation of trisaccharide intermediate III-1

[0094] 1. Preparation method of monosaccharide intermediate 3-1

[0095] Intermediate 9 was obtained by reacting glucosamine hydrochloride 8 with benzyloxyformyl chloride; 1,6-cyclic intermediate 10 was obtained through a two-step reaction; catalytic hydrogenation and azo transfer reaction were used to obtain a common intermediate 11 with an azido group protected at the 2-position; the hydroxyl groups at the 3- and 4-positions were fully benzylated under the action of benzyl bromide and sodium hydride to obtain intermediate 12; 1,6-cyclic intermediate 13 was obtained under the action of acetic anhydride and TBSOTf; the terminal acetyl group was removed under the action of benzylamine to obtain intermediate 14; finally, trichloroacetylimine ester donor 3-1 was obtained under the action of potassium carbonate and trichloroacetonitrile.

[0096] The reaction conditions and yields for each step are as follows: a) Cb2Cl, NaOH, H2O, 59%; b) 1) TsCl, Py, MS, 2) EtOH, DBU, NaI, two-step yield 49%; c) 1) 4atm H2, Pd / C, MeOH, 2) TfN3, CuSO4, Et3N, MeOH, two-step yield 82%; d) BnBr, NaH, DMF, 0℃, 85%; e) Ac2O, TBSOTf, 94%; f) BnNH2, DCM, 89%; g) Cl3CCN, K2CO3, DCM, rt, 98%.

[0097] 2. Preparation method of fully protected trisaccharide intermediate 1-1

[0098] The synthesized monosaccharide intermediate 3-1, in its mixed configuration, can be used directly in the next reaction without separation. Trichloroacetylimine ester is a relatively common glycosyl donor, typically reacting under mild conditions with high yields. Monosaccharide intermediate 3-1 is glycosylated and coupled with disaccharide intermediate 4-1 synthesized according to known literature methods, yielding a fully protected trisaccharide 1-1 in the presence of trifluoromethanesulfonic acid.

[0099] Monosaccharide intermediate 3-1 (2.83 g, 502.24 mmol) and glycosyl acceptor disaccharide intermediate 4-1 (2.74 g, 324.23 mmol) were dissolved in redistilled DCM and then added to a solution containing pre-activated DCM. In a reaction flask containing molecular sieves, the mixture was continuously stirred at room temperature for 30 min to equilibrate the reaction. After the reaction system temperature was lowered to -20℃, trifluoromethanesulfonic acid (26.53 μL, 0.33 mmol) was slowly added dropwise. The reaction was monitored by TLC. After the reaction was complete, the molecular sieves were removed by silica gel filtration. The resulting filtrate was concentrated and purified by direct silica gel column chromatography (PE / EA = 5:1) to obtain the fully protected trisaccharide intermediate 1-1 (3.42 g, 87%).

[0100] 1H NMR(400MHz,CDCl3)δ8.08(d,J=7.6Hz,2H),7.41–7.25(m,22H),7.21–7.18(m,6H),5.46(d,J=3.1Hz,1H),5.14(t,J=3.6Hz,1H),5.02(d,J=2.2Hz,2H),4.88(s,1H),4.85(d,J=3.0Hz,2H),4.82(d,J=5.3Hz,2H),4.77(d,J=4.2Hz,1H),4.74(d,J=3.8Hz,1H),4.64(d,J=3.8Hz,1H),4.58(d,J=10.4Hz,1H),4.54(d,J=10.9Hz,1H),4.32(d,J=2.7Hz,1H),4.30(d,J=2.2Hz,3H),4.24(d,J=3.6Hz,1H),4.21(d,J=3.9Hz,1H),4.14(t,J=4.3Hz,1H),4.02(t,J=4.0Hz,1H),3.99–3.90(m,2H),3.73(dt,J=9.9Hz,3.2Hz,1H),3.63(d,J=3.8Hz,1H),3.61(d,J=2.8 1H),3.58(s,3H),3.47(t,J=9.4Hz,1H),3.31(s,3H),3.21(dd,J=10.2,3.5Hz,1H).

[0101] 13 C NMR(100MHz,CDCl3)δ170.75,170.52,169.35,165.42,155.81,138.23,137.69,137.58,137.42,136.24,133.37,130.01,129.38,128.74,128.52,128.49,128.44,128.39,128.24,128.17,128.13,128.06,127.96,127.88,127.76,127.35,99.32,98.87,98.50,80.05,79.12,77.29,75.62,75.28,74.98,74.90,74.56,74.44,73.32,70.07,69.33,69.12,68.92,67.00,63.62,62.45,62.33,55.29,54.56,52.10,20.87.

[0102] HRMS[M+Na] + m / z 1275.46443(calcd for C67 H 72 N4NaO 20 ,1275.4638).

[0103] 3. Preparation method of trisaccharide intermediate III-1

[0104] The fully protected trisaccharide 1-1 was subjected to the combined action of LiOH, H2O2 and NaOH to simultaneously remove Ac, Bz and methyl ester to obtain trihydroxy compound I-1; heating under the action of SO3·NMe3 gave the intermediate compound II-1 after O-sulfonation; benzyl and Cbz were removed by catalytic hydrogenation, and the azide was reduced to generate an amino group to obtain diamino compound III-1.

[0105] The fully protected trisaccharide compound 1-1 (342.62 mg, 0.26 mmol) was dissolved in 5.00 mL of tetrahydrofuran. Then, 6.23 mL of 1.25 N LiOH solution and 13.13 mL of 30% H₂O₂ solution were added dropwise at room temperature. After stirring for 12 hours, 14.34 mL of methanol and 7.82 mL of 6 N NaOH solution were added, and stirring continued for at least 12 hours. After the reaction was confirmed to be complete by TLC, the pH was adjusted to 2 with 4 N hydrochloric acid under ice-water bath conditions. The reaction solution was then extracted three times with DCM and concentrated under reduced pressure, followed by silica gel column chromatography (DCM:MeOH = 15:1) to obtain compound I-1 (258.0 mg, 93%). High-performance liquid chromatography analysis of intermediate I-1 revealed only a single peak, indicating its high purity. Under argon protection, intermediate I-1 (258.02 mg, 0.24 mmol) and SO3·NMe3 (897.62 mg, 5.33 mmol) were dissolved in 3 mL of anhydrous DMF. The reaction system was heated to 65 °C and stirred continuously for at least 12 h. The reaction solution was taken and the degree of reaction was monitored by high performance liquid chromatography. The reaction was compared with the peak time of compound I-1. When the newly generated peak had a shorter retention time and was a single peak, the reaction was considered complete. After heating was stopped, the reaction system was allowed to rise naturally to room temperature. The reaction solution was concentrated and purified by Sephadex LH-20 gel column chromatography to obtain intermediate II-1 (304.13 mg, 93%). Intermediate II-1 (304.13 mg, 0.22 mmol) was dissolved in 3.00 mL of a mixed solvent of methanol, tert-butanol and water (v / v / v = 2:1:1), and palladium on carbon (50.00 mg) was added. The mixture was stirred for 24 h under a hydrogen pressure of 4 atm. After removing the palladium on carbon by filtration with filter paper, the reaction solution was concentrated to obtain intermediate III-1 (184.93 mg, 98%).

[0106] 1H NMR(400MHz,D2O)δ5.33(d,J=3.7Hz,1H),5.15(s,1H),4.88(d,J=3.7Hz,1H),4.79(d,J=1.8Hz,1 H),4.32–4.25(m,3H),4.23(d,J=3.1Hz,2H),4.13(d,J=2.1Hz,1H),4.11–4.06(m,2H),3.95(d,J =5.6Hz,1H),3.85(t,J=9.8Hz,2H),3.77(t,J=9.9Hz,1H),3.69(d,J=9.5Hz,1H),3.65(s,1H),3. 47(t,J=9.7Hz,1H),3.35(s,3H),3.26(dd,J=10.7Hz,3.6Hz,1H),3.22(dd,J=10.5Hz,3.7Hz,1H).

[0107] 13 C NMR(100MHz,D2O)δ175.15,98.88,96.32,91.44,76.78,73.00,70.50,70.26, 69.39,69.19,68.94,68.67,67.32,66.61,66.16,62.96,55.32,54.22,54.03.

[0108] HRMS[M–H] - m / z 769.0596 (calcd for C 19 H 33 N2O 24 S3,769.0586).

[0109] Example 2: Preparation of compound CV010

[0110] Intermediate III-1 (35 mg, 0.045 mmol) was dissolved in 0.50 mL of water, and the pH was adjusted to 9-10 with 4N NaOH solution and maintained. Then, sulfur trioxide pyridine complex (216 mg, 1.366 mmol) was added in portions. The reaction was confirmed to be complete by TLC. The pH of the neutralized reaction solution was approximately 7-8. The reaction solution was then concentrated and purified by Sephadex G-25 gel column chromatography to obtain compound CV010 (39 mg, 94%).

[0111] 1H NMR(400MHz,D2O)δ5.37(d,J=3.6Hz,1H),5.18(d,J=3.0Hz,1H),4.97(d,J=3.6Hz,1H),4.78 (d,J=2.8Hz,1H),4.33-4.26(m,4H),4.21–4.14(m,2H),4.06(t,J=3.2Hz,1H),3.92(td,J=8. 0Hz,7.5Hz,3.5Hz,2H),3.70(t,J=9.5Hz,1H),3.64(d,J=10.1Hz,1H),3.59(d,J=10.1Hz,1H ),3.52(t,J=9.5Hz,1H),3.37(s,3H),3.35(s,1H),3.22(ddd,J=13.1Hz,10.1Hz,3.6Hz,2H).

[0112] 13 C NMR(100MHz,D2O)δ175.37,99.29,98.32,96.95,76.88,75.94,75.90,70.98, 70.05,69.87,69.24,69.15,68.94,68.56,66.87,66.49,57.92,57.76,55.43.

[0113] HRMS[M–H] - m / z 928.9717 (calcd for C 19 H 33 N2O 30 S5,928.9722).

[0114] Example 3 Preparation of compound CV016

[0115] Compound CV010 (39.4 mg, 0.042 mmol) was treated with Dowex-50-WX4-Na + The column exchanged the sodium salt, and the sugar-containing component was collected to concentrate the solvent, yielding compound CV016 (44 mg, 98%).

[0116] 1H NMR(400MHz,D2O)δ5.35(d,J=3.6Hz,1H),5.16(d,J=3.0Hz,1H),4.95(d,J=3.6Hz,1H),4.75(d,J =2.8Hz,1H),4.27(dq,J=13.0Hz,3.8Hz,2.9Hz,4H),4.19–4.12(m,2H),4.04(t,J=3.2Hz,1H),3.9 0(td,J=8.0Hz,7.5Hz,3.5Hz,2H),3.68(t,J=9.5Hz,1H),3.61(d,J=10.1Hz,1H),3.56(d,J=10.1H z,1H),3.50(t,J=9.5Hz,1H),3.35(s,3H),3.33(s,1H),3.20(ddd,J=13.1Hz,10.1Hz,3.6Hz,2H).

[0117] 13 C NMR(100MHz,D2O)δ174.36,99.23,98.22,96.93,76.84,75.93,75.89,70.96, 70.01,69.84,69.22,69.13,68.91,68.51,66.84,66.46,57.89,57.73,55.40.

[0118] HRMS[M-6Na+5H] - m / z 928.9754 (calcd for C 19 H 33 N2O 30 S5,928.9722).

[0119] Example 4: Preparation of fully protected trisaccharide intermediate 2

[0120] 1. Preparation of monosaccharide intermediate F-1

[0121] Compound 17 was deacetylated under alkaline conditions of MeOH / K2CO3, followed by the addition of a benzylidene reagent and adjustment to acidity with p-toluenesulfonic acid monohydrate to give intermediate 18. Intermediate 18 was refluxed with dibutyltin oxide in anhydrous methanol, and then cesium fluoride and 4-methoxybenzyl chloride were added to selectively protect the 3-hydroxyl group to give intermediate 19. Compound 19 was reacted with benzyl bromide under neutral silver oxide conditions to give compound 20. Compound 20 was deprotected from the PMB protecting group in the presence of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone to give compound 21. Compound 22 was given by adding triethylamine and acetic anhydride dropwise under DMAP catalysis using dichloromethane as solvent. Compound 22 was dissolved in dichloromethane, and the benzylidene group was removed by adding trifluoroacetic acid to give compound 23. Under ice bath conditions, an equal amount of benzoyl chloride was added to selectively protect the 6-position, and the intermediate was purified. The benzylidene group was then moved to the 4-position under silver oxide conditions to give the monosaccharide compound F-1. The reaction process is as follows:

[0122] a) 1) K2CO3, MeOH, 25℃-rt; 2) Benzaldehyde, Dimethylacetal, Toluene-4-sulfonic acid, DMF, two-step yield 78%;

[0123] b) 1) Bu₂SnO, MeOH, reflux; 2) PMBCl, CsF, DMF, 90℃, two-step yield 67%; c) Ag₂O, BnBr, DCM, 88%; d) DDQ, DCM, H₂O, 71%; e) Et₃N, Ac₂O, DMAP, DCM, 90%; f) TFA, DCM, 79%; g) 1) BzCl, Et₃N, THF, 0℃-rt; 2) Ag₂O, BnBr, DCM, two-step yield 72%.

[0124] The specific steps are as follows: Dissolve 10g of compound 17 (30.21mmol) in methanol, add 12.51g of potassium carbonate (90.63mmol) in portions, heat to 25℃, monitor the reaction by TLC until a new spot is formed, filter, and directly add the crude product to the next step. Dissolve the obtained solid in DMF, place on a stirrer, add 9.06ml of benzaldehyde dimethyl acetal (60.42mmol), and adjust the pH to 2-3 with p-toluenesulfonic acid monohydrate. React overnight. The next day, the starting material disappears on the TLC, stop the reaction, and quench with triethylamine. After drying the solvent, extract with DCM and saturated NaCl system, collect the organic phase, dry, and separate by column chromatography to obtain white solid 18 (5.92g, two-step reaction yield 78%).

[0125] 1H NMR (400MHz, CDCl3) δ7.49-7.42(m,4H),7.36(m,3H),7.14(m,2H),5.50(s,1H),4.54(d,J=9.7Hz, 1H), 4.35 (dd, J = 10.4, 3.9Hz, 1H), 3.83-3.73 (m, 2H), 3.49-3.38 (m, 3H), 2.92 (s, 1H), 2.36 (s, 3H).

[0126] 13 C NMR (100MHz, CDCl3) δ138.95,137.02,133.78,130.01,129.44,128.48,127. 44,126.43,102.02,88.80,80.35,74.65,72.62,70.62,68.70,29.81,21.30.

[0127] 10 g of compound 18 (26.73 mmol) and 9.95 g of dibutyltin oxide (40.107 mmol) were dissolved in 30 mL of anhydrous methanol. The mixture was placed on a magnetic stirrer and refluxed at 60 °C for 6 h. The reaction system slowly changed from a suspension to a clear liquid. After the reaction solution was slowly cooled to room temperature, it was concentrated under reduced pressure to remove the anhydrous methanol. Without further purification, the reactants were directly dissolved in 20 mL of dry DMF. 7.22 mL of p-methoxybenzyl chloride (53.47 mmol) and... 4.87 g of cesium fluoride (32.086 mmol) was added to the reaction system and refluxed and stirred overnight at 90 °C. After the reaction of the starting materials was completely eliminated by TLC monitoring, the reaction solution was slowly cooled to room temperature and concentrated under reduced pressure using an oil pump to remove the reaction solvent DMF. The resulting oily substance was redissolved with EA, and then the reaction solution was extracted with saturated NaCl solution. The extraction was performed three times in the forward direction and twice in the reverse direction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate hydrate, and concentrated under reduced pressure using a water pump. Compound 19 (9.906 g, 67%) was obtained by column chromatography.

[0128] 1H NMR(400MHz, CDCl3)δ7.49(m,2H),7.43(m,2H),7.40-7.36(m,3H),7.28(m ,2H),7.13(m,2H),6.87-6.83(m,2H),5.56(s,1H),4.88(d,J=11.1Hz,1H) ,4.71(d,J=11.1Hz,1H),4.56(d,J=9.6Hz,1H),4.38(dd,J=10.4,4.9Hz,1 H),3.79(m,4H),3.64(m,2H),3.51-3.44(m,2H),2.55(s,1H),2.35(s,3H).

[0129] 13 C NMR (100MHz, CDCl3) δ159.52,138.87,137.35,133.97,130.39,129.94,129.14,128.39,127.33, 126.13,114.02,101.35,88.61,81.32,81.28,77.36,74.59,72.14,70.86,68.76,55.38,21.30.

[0130] 9.90 g of compound 19 (26.68 mmol) was dissolved in redistilled dichloromethane. 18.51 g of silver oxide (80.04 mmol) was rapidly weighed in portions and added to the reaction system. The system was cooled to 0 °C in an ice bath. 9.63 mL of benzyl bromide (80.04 mmol) was added dropwise. After the reaction of the starting material was completely eliminated by TLC monitoring, the silver oxide was filtered off, the organic phase was concentrated, and separated by column chromatography to obtain compound 20 (10.82 g, yield 88%).

[0131] 1 H NMR (400MHz, CDCl3) δ7.55-7.30(m,14H),7.12(m,2H),6.82(m,2H),5.59(s,1H),4.86(m,3H),4.71(t,J=10.0 Hz,2H),4.39(dd,J=10.6,5.0Hz,1H),3.80(d,J=6.0Hz,5H),3.67(t,J=9.4Hz,1H),3.46(m,2H),2.35(s,3H).

[0132] 13C NMR (100MHz, CDCl3) δ159.44,138.30,137.44,133.17,130.59,129.95,129.89,129.25,129.09,128.52,128.38,128.29 ,127.96,126.13,113.93,101.24,88.60,82.81,81.62,80.49,77.36,75.92,75.09,70.35,68.85,55.38,29.82,21.26.

[0133] 4.43 g of compound 20 (9.62 mmol) was dissolved in 30 mL of a 10:1 mixture of dichloromethane and water. 3.27 g of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (14.44 mmol) was weighed and added to the reaction flask. The reaction was carried out at room temperature for 3 h. After the starting material spot disappeared completely as monitored by TLC, the red precipitate formed in the reaction solution was filtered off. DCM was then added to the reaction system, and the mixture was extracted with DCM and saturated NaCl solution. The mixture was dried over anhydrous sodium sulfate, filtered, and separated by column chromatography to obtain compound 21 (2.34 g, 71%).

[0134] 1 H NMR (400MHz, CDCl3) δ7.49-7.34(m,12H),7.15(m,2H),5.52(s,1H),4.95(d,J=10.9Hz,1H),4.82(d,J=10.9Hz,1H),4.69(d,J=9.7Hz,1H),4. 36(dd,J=10.5,4.9Hz,1H),3.90(t,J=8.8Hz,1H),3.77(t,J=10.1Hz,1H),3.50(d,J=9.3Hz,1H),3.46–3.39(m,2H),2.50(s,1H),2.37(s,3H).

[0135] 13 C NMR (100MHz, CDCl3) δ138.32,138.22,137.09,133.01,132.52,129.95,129.92,129.87,129.38,129.32,129.11,128.76, 128.67,128.45,128.34,128.27,128.15,126.40,101.90,88.33,80.87,80.41,77.36,75.56,75.53,70.15,68.77,21.27.

[0136] 2.34 g of compound 21 (6.86 mmol) was dissolved in dichloromethane. An ice bath was added, and the reaction flask was fixed to a stirrer. 2.85 mL of triethylamine (20.58 mmol), 1.93 mL of acetic anhydride (20.58 mmol), and a catalyst amount of dimethylaminopyridine were added. After the addition was complete, the ice bath was removed. After 3 hours, the mixture was spotted onto a TLC plate. The starting material disappeared, and a slightly less polar compound was formed. The reaction was then stopped. The solution was evaporated to dryness and purified by column chromatography to give a white solid 22 (2.3 g, 90%).

[0137] 1 H NMR (400MHz, CDCl3) δ7.47-7.42(m,4H),7.36-7.32(m,8H),7.15(m,2H),5.47(s,1H),5.39(t,J=9.2Hz,1H),4.93(d,J=11.0Hz,1H),4.77( d,J=9.7Hz,1H),4.64(d,J=10.9Hz,1H),4.37(dd,J=10.5,4.8Hz,1H),3.79(d,J=10.2Hz,1H),3.61-3.51(m,3H),2.37(s,3H),1.96(s,3H).

[0138] 13 C NMR (100MHz, CDCl3) δ169.87,138.52,137.79,137.04,133.12,130.02,129.18,128.60,128.36,128 .27,128.07,126.27,101.47,88.86,79.32,78.71,77.36,75.32,74.45,70.45,68.75,21.32,21.04.

[0139] 2.3 g (6.00 mmol) of compound 22 was dissolved in dichloromethane solution, and 9.8 mL of trifluoroacetic acid (60.00 mmol) was slowly added. The reaction was stopped when the white solid disappeared and the reaction solution became clear. The solution was adjusted to neutral with sodium bicarbonate, and then extracted with EA and saturated NaCl. The organic phase was collected, dried over anhydrous sodium sulfate, filtered and evaporated to dryness, and purified by column chromatography to give white solid 23 (1.41 g, 79%).

[0140] 1H NMR (400MHz, CDCl3) δ7.45-7.41(m,2H),7.36-7.28(m,5H),7.13(m,2H),5.03(t,J =9.2Hz,1H),4.89(d,J=11.0Hz,1H),4.68(d,J=9.8Hz,1H),4.59(d,J=11.0Hz,1H) ,3.89(dd,J=12.0,3.2Hz,1H),3.78(dd,J=12.0,4.8Hz,1H),3.59(t,J=9.6Hz,1H) ,3.45(t,J=9.3Hz,1H),3.39-3.33(m,1H),2.66(s,2H),2.34(s,3H),1.99(s,3H).

[0141] 13 C NMR (100MHz, CDCl3) δ172.15,138.31,137.87,132.78,129.97,129.26,128.56, 128.10,128.02,87.82,79.70,79.37,78.62,75.25,69.92,62.48,21.24,21.08.

[0142] 1.41 g of compound 23 (4.78 mmol) was dissolved in dry tetrahydrofuran. Under ice bath conditions, 0.73 mL of triethylamine (5.26 mmol) was added, followed by the dropwise addition of 0.61 mL of benzoyl chloride (5.26 mmol). After a new spot was observed by TLC, the triethylamine was neutralized with acidic resin, filtered, and evaporated to dryness to obtain a crude product with Bz at position 6, which was directly used in the next step. This intermediate was dissolved in redistilled dichloromethane, and 3.3 g of silver oxide (14.34 mmol) was rapidly weighed in portions and added to the reaction system. The system was cooled to 0 °C in an ice bath, and 1.7 mL of benzyl bromide (14.34 mmol) was added dropwise. After the reaction of the starting material was completely eliminated by TLC, the silver oxide was filtered off, the organic phase was concentrated, and separated by column chromatography to obtain compound F-1 (1.68 g, two-step yield 72%).

[0143] 1H NMR (400MHz, CDCl3) δ8.07-8.01(m,2H),7.62(d,J=7.4Hz,1H),7.46(dd,J=7.8,6.6Hz,5H),7.34-7.25 (m,7H),7.22(dd,J=8.2,6.2Hz,2H),6.94(dd,J=8.2,6.1Hz,2H),5.35(d,J=9.2Hz,1H),4.90(d,J=11. 0Hz,1H),4.65(dd,J=16.7,5.9Hz,2H),4.55(dd,J=11.3,3.1Hz,3H),4.46(dd,J=12.0,5.4Hz,1H),3.7 2(ddd,J=9.6,5.3,2.0Hz,1H),3.60(d,J=9.5Hz,1H),3.45(d,J=9.4Hz,1H),2.27(s,3H),1.87(s,3H).

[0144] 13 C NMR (100MHz, CDCl3) δ169.79,166.09,138.09,137.80,137.18,133.17,129.95,129.84,129.70,128.94,128.58,128.48,12 8.44,128.33,128.13,128.09,127.91,87.47,78.74,77.41,77.09,76.91,76.77,76.26,74.76,74.59,63.50,21.18,21.02.

[0145] 2. Preparation method of fully protected trisaccharide intermediate 2

[0146] The synthesized monosaccharide intermediate F-1 was glycosylated and coupled with the disaccharide intermediate 4-1 synthesized according to the method in the known literature, and a fully protected trisaccharide 2 was generated under the action of trimethylsilyl ester.

[0147] Preparations before the reaction: Use a microwave oven to bake the molecular sieve to remove water, and put the two-necked flask and magnetic flask into an oven to dry.

[0148] Remove the two flasks after baking for half an hour, put the magnetic jar in your pocket, and quickly add the baked contents. Molecular sieve (2.5001g), connected to a double-row tube, and the oil pump turned on, the strong suction force securely connects the bottle, ensuring the airtightness of the apparatus. Then, heat the reaction flask with a blowtorch, continuously shaking and tapping the flask until the molecular sieve becomes soft, smooth, and non-sticky to the walls. Close the oil pump valve, insert a magnetic stopper, plug the flask, open the oil pump valve, and continue heating with the blowtorch for 10 seconds. Fix the reaction flask on the iron stand, open the argon gas valve, and perform purging three times. After purging, turn off the oil pump; the pre-reaction preparation is complete. Dissolve 100mg of donor F-1 (0.20mmol) and 173mg of acceptor 4-1 (0.20mmol) in redistilled dichloromethane, slowly inject into the reaction flask using a syringe, turn on the stirrer, and equilibrate for 40 minutes. Anhydrous ethanol and liquid nitrogen were poured into a Dewar flask and adjusted to -40°C. The reaction flask was cooled, and after 15 minutes, 138 mg of N-iodosuccinimide (0.60 mmol) was added. A catalyst amount of trimethylsilyl trifluoromethanesulfonate was added using a microsyringe. The mixture was continuously cooled, and after 1 hour, the reaction was complete on TLC. Triethylamine was added to quench the reaction, the argon gas was turned off, and post-processing was performed. The mixture was filtered through a funnel containing a section of silica gel, washed repeatedly with ethyl acetate, concentrated, and purified by column chromatography to obtain the fully protected trisaccharide compound 2 (183 mg, 69%).

[0149] 1 H NMR (400MHz, CDCl3) δ8.06-7.94(m,4H),7.59-7.33(m,7H),7.29(m,6H),7.25-7.19(m,11H),7.17-7.08(m,5H),7.08-7 .03(m,2H),5.56-5.44(m,2H),5.21(t,J=5.3Hz,1H),5.04(m,2H),4.85-4.71(m,3H),4.64(s,3H),4.60-4.51(m,4H),4. 44(dd,J=12.2,3.5Hz,1H),4.38(d,J=12.4Hz,1H),4.31(dd,J=12.3,3.6Hz,1H),4.23(m,2H),4.18-4.06(m,3H),4.02- 3.89(m,2H),3.72-3.62(m,4H),3.62-3.52(m,2H),3.38(dd,J=10.1,3.5Hz,1H),3.26(s,3H),2.10(s,3H),1.89(s,3H).

[0150] 13C NMR (100MHz, CDCl3) δ170.95,169.98,169.67,166.19,165.67,155.89,138.49,137.90,137.72,137.60,136. 40,133.25,130.18,129.96,129.72,129.27,128.58,128.53,128.43,128.30,128.26,128.23,128.01,127.97 ,127.85,127.60,127.52,127.40,98.92,98.70,98.43,78.49,77.36,76.56,76.41,76.26,75.52,74.51,74.41,73.52,73.22,72.69,71.12,70.99,69.80,68.99,66.98,63.11,62.22,55.33,54.44,52.34,21.14,20.95.

[0151] Example 5: Preparation of compound TSC19

[0152] Deprotection: 90 mg of the fully protected trisaccharide 2 was dissolved in an appropriate amount of tetrahydrofuran, and 1 mL of 1.25 N LiOH solution and 2.1 mL of 30% H₂O₂ solution were added. The reaction flask was fixed on a stirrer, and after 12 h, 2.2 mL of anhydrous methanol and 1.2 mL of 4 N NaOH solution were added, and the reaction was continued for another 12 h. After observing the formation of a thin and long product spot on the TLC plate, the pH of the reaction solution was adjusted to 5–6 with 6 N HCl. Extraction was performed using a DCM and saturated NaCl solution system, and the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Column chromatography was performed (DCM:MeOH = 15:1 → 10:1) to obtain the deprotected compound (40 mg, 58%).

[0153] Oxysulfonic acidification: Before the reaction, place the 25 mL flask and magnetic flask to be used in an oven and bake for 30 min to keep them dry. Weigh 220 mg SO3·NMe3 (1.58 mmol) according to 20 equivalents for each hydroxyl group and add it to the flask. Seal the flask tightly. Dissolve 20 mg of the deprotected trisaccharide compound (0.015 mmol) in as little anhydrous DMF as possible and add the solution dropwise to the reaction flask using a syringe. Stir the reaction continuously at 60 °C for 36 h, then evaporate the solvent. Redissolve the oily substance in methanol and perform column chromatography using a Sephadex LH-20 gel column with a 1:1 volume ratio of dichloromethane and methanol as the eluent. Collect the sugar-containing fraction, evaporate the solvent, and elute with methanol. Filter the solution through a Dowex-50-WX4-Na gel column. +The column was used to exchange the sugar component for sodium salt, and the sugar-containing component was collected and concentrated to obtain an oxosulfonated compound (24 mg, 87%).

[0154] Hydrogenation and nitrogen sulfonation: Dissolve 24 mg of the oxysulfonation compound (0.017 mmol) in 3 mL of methanol and add an equal volume of H2O. Then weigh 20 mg of palladium on carbon and place it in a reaction flask. React for 24 h in a hydrogen atmosphere at 4 atm. Filter off the palladium on carbon using an organic phase filter membrane. The solution is colorless. Dry the filtrate by rotary evaporation. NMR shows no peaks in the aromatic region. Then proceed to the next step. The hydrogenated product was dissolved in 4 mL of H₂O. 2N NaOH solution was added to adjust the pH of the reaction solution to 9–10. SO₃·Py (53 mg, 0.33 mmol) was weighed and divided into four equal portions, added to the reaction solution at 30-minute intervals. After addition, the mixture was stirred at room temperature for 4 hours. Throughout the addition and reaction process, the pH of the reaction solution was constantly monitored to maintain it between 9 and 10. After the reaction time was reached, the pH was neutralized to 7–8 with hydrochloric acid, followed by Sephadex G-25 gel column chromatography using H₂O as the eluent. The sugar-containing fraction was collected, and then further eluted with H₂O using a Dowex-50-WX4-Na₂O gel column chromatography. + The column was exchanged to exchange it for sodium salt, the sugar-containing component was collected, the solvent was concentrated, and the final compound TSC19 (15 mg, two-step yield 84%) was obtained.

[0155] 1 H NMR(400MHz,D2O)δ5.26(d,J=3.8Hz,1H),5.17(s,1H),5.01(d,J=3.5Hz,1H),4.43(t,J=9.7Hz,1H),4.33(m,3H),4.27(s,1H),4.21-4.18(m, 1H),4.16-4.12(m,1H),3.99(m,3H),3.82-3.75(m,1H),3.73(d,J=9.6Hz,1H),3.71-3.62(m,3H),3.41(s,3H),3.27(dd,J=10.2,3.6Hz,1H).

[0156] 13 C NMR(100MHz,D2O)δ174.96,99.21,98.13,94.97,81.69,77.15,74.21,72.02, 69.89,69.79,69.64,68.45,68.13,67.40,66.76,66.20,65.18,57.73,55.32.

[0157] Example 6: Therapeutic effect of CV016 / TCS19 in a mouse model of COPD

[0158] 1.1 Establishment of a mouse COPD model (LPS+CS)

[0159] Experimental groups: control group (blank group), model group, CV016 group, TSC19 group

[0160] Except for the control group, mice were placed in a self-made 60L sealed smoke chamber with a three-way valve. Except for the control group, all other groups were passively exposed to smoke once a day for one hour after administration, for 70 consecutive days. On days 1 and 15, mice were not exposed to smoke. After intraperitoneal injection of 4% sodium pentobarbital for anesthesia, mice were given 0.2 ml of 1 mg / mL lipopolysaccharide solution via tracheal infusion. Once the mice regained consciousness, they were returned to their original enclosures for continued care. During the exposure period, the weight and other data of each mouse were continuously observed and recorded. In the blank control group, 0.2 ml of physiological saline was administered via tracheal infusion after anesthesia at the corresponding lipopolysaccharide exposure time points. Except for the control group and the CS / LPS model group, which received an equal volume of distilled water, the CV016 and TSC19 groups were subcutaneously administered CV016 or TSC19 20 mg / kg once daily in the morning, starting from the day of modeling and continuing for 70 consecutive days until the day modeling was completed.

[0161] 1.2 Detection Indicators

[0162] The left lung of anesthetized mice was lavaged three times with pre-cooled phosphate-buffered saline (PBS, pH 7.4) three times, 0.5 ml each time. The collected fluid was centrifuged at 4°C and 1500×g for 15 min, and the supernatant was the mouse bronchoalveolar lavage fluid (BALF). Then, 200 mg of mouse left lung tissue was weighed, added to 200 μL of PBS solution, and homogenized until no visible debris was found. The tissue was then centrifuged at 4°C and 10,000×g for 15 min, and the lung tissue supernatant was collected. The levels of inflammatory factors TNF-α, IL-6, and I-1β in the bronchoalveolar lavage fluid were detected according to the ELISA kit instructions.

[0163] Tissue from the upper lobe of the right lung of unperfused mice was collected, fixed in 10% formalin solution, and then dehydrated with different concentrations of ethanol at room temperature before being embedded in paraffin. The embedded lung tissue was cut into 4 μm thick sections using a microtome. Following the procedures of the hematoxylin and eosin (H&E) staining kit and the Masson trichrome staining kit, the lung tissue sections were stained with H&E and Masson staining, and observed and photographed under an optical microscope.

[0164] 1.3 Experimental Results

[0165] The weight gain of mice in each group did not differ significantly during the experiment.

[0166] 1.3.1 CV016 / TCS19 can downregulate cytokine levels in mouse BALF.

[0167] To verify the effects of CV016 / TSC19 on airway inflammation in mice, we used ELISA to detect the expression levels of IL-1β, IL-6, and TNF-α in the bronchoalveolar lavage fluid (BALF) of mice in the control group (Control), model group (COPD), CV016 group (COPD+CV016), and TSC19 group (COPD+TSC19). The results showed that the expression levels of IL-1β, IL-6, and TNF-α in the COPD group were significantly higher than those in the Control group (P<0.05); compared with the COPD group, the expression of IL-1β, IL-6, and TNF-α in the COPD+CV016 and COPD+TSC19 groups was significantly reduced (P<0.05).

[0168] 1.3.2 CV016 / TCS19 alleviates pathological damage in lung tissue of COPD mice

[0169] As shown by H&E staining results, the control group exhibited intact bronchial structures, clear alveolar structures, and minimal surrounding inflammatory cell infiltration. The COPD group showed narrowed bronchial lumens, alveolar and lung tissue structure destruction, epithelial cell shedding, exfoliated cells within the alveolar cavities, and extensive peribronchial and perivascular inflammatory cell infiltration. Compared to the COPD group, the CV016 and TCS19 groups showed significantly reduced lung tissue structural damage, decreased bronchial mucosal edema, and reduced inflammatory cell infiltration, with the CV016 group exhibiting significantly more normalized characteristics.

[0170] 1.3.3 CV016 / TSC19 reduces increased peri-airway collagen deposition in the lung tissue of COPD mice

[0171] Airway collagen deposition induces airway obstruction, which is a pathological feature of COPD. Masson staining of mouse lung sections showed that, compared with the COPD group, the CV016 and TSC19 groups of mice had reduced collagen deposition in the airways.

[0172] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Use of a trisaccharide compound of formula A, or a salt thereof, or a solvate thereof, in the preparation of a medicament for the treatment or prevention of chronic obstructive pulmonary disease (COPD): in, R1, R2, R3, and R4 may be the same or different, and are independently selected from H or -SO3H; R5 is -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H; R6 is -OH, -OSO3H, or -NHSO3H. The salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation; The monovalent cation is selected from Na. + K + Li + NH4 + The divalent cation is selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ The trivalent cation is selected from Al. 3+ Fe 3+ .

2. Compounds of formula A with a trisaccharide structure, or salts thereof, or solvates thereof, are used for the treatment or prevention of chronic obstructive pulmonary disease (COPD): in, R1, R2, R3, and R4 may be the same or different, and are independently selected from H or -SO3H; R5 is -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H; R6 is -OH, -OSO3H, or -NHSO3H. The salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation; The monovalent cation is selected from Na. + K + Li + NH4 + The divalent cation is selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ The trivalent cation is selected from Al. 3+ Fe 3+ .

3. A method for treating or preventing chronic obstructive pulmonary disease (COPD), characterized in that, Administering a therapeutically effective amount of a compound of formula A or a salt thereof or a solvation thereof, or a pharmaceutical composition containing a compound of formula A or a salt thereof or a solvation thereof, to a patient in need. Among them, R1, R2, R3 and R4 are the same or different, and are independently selected from H or -SO3H; R5 is -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H; R6 is -OH, -OSO3H or -NHSO3H; The salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation; The monovalent cation is selected from Na. + K + Li + NH4 + The divalent cation is selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ The trivalent cation is selected from Al. 3+ Fe 3+ .

4. Treatment or prevention of COPD using a compound of formula A or a salt thereof or a solvate thereof, or using a pharmaceutical composition containing a compound of formula A or a salt thereof or a solvate thereof. in, R1, R2, R3, and R4 may be the same or different, and are independently selected from H or -SO3H; R5 is -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H; R6 is -OH, -OSO3H, or -NHSO3H. The salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation; The monovalent cation is selected from Na. + K + Li + NH4 + The divalent cation is selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ The trivalent cation is selected from Al. 3+ Fe 3+ .

5. When treating or preventing COPD, the use of a compound of formula A or a salt thereof or a solvation thereof, or the use of a pharmaceutical composition containing a compound of formula A or a salt thereof or a solvation thereof, in, R1, R2, R3, and R4 may be the same or different, and are independently selected from H or -SO3H; R5 is -C1-5 alkyl, -C1-5 alkylene-NH2, or -C1-5 alkylene-NHSO3H; R6 is -OH, -OSO3H, or -NHSO3H. The salt is a salt formed with a monovalent cation, a divalent cation, and / or a trivalent cation; The monovalent cation is selected from Na. + K + Li + NH4 + The divalent cation is selected from Ca. 2+ Cu 2+ Zn 2+ Fe 2+ Mg 2+ Mn 2+ The trivalent cation is selected from Al. 3+ Fe 3+ .

6. The use or method as described in any one of claims 1-5, characterized in that, In formula A, R1 is selected from H or -SO3H; R2, R3 and R4 are the same, all of which are H; R6 is -OH, -OSO3H or -NHSO3H.

7. The use or method as described in any one of claims 1-5, characterized in that, In formula A, R1, R2, R3 and R4 are all H, and R6 is -NHSO3H.

8. The use or method as described in any one of claims 1-5, characterized in that, In formula A, R1 is -SO3H, R2, R3 and R4 are the same, all are H, and R6 is -OH.

9. The use or method as described in any one of claims 1-8, characterized in that, In formula A, R5 is -C1-3 alkyl, -C2-4 alkylene-NH2, or -C2-4 alkylene-NHSO3H; Preferably, R5 is methyl, ethyl, -C2H4NH2, -C2H4NHSO3H, -C3H6NH2 or -C3H6NHSO3H.

10. The use or method as described in any one of claims 1-5, characterized in that, The compound or its salts are selected from compounds with the following structures:

11. The use or method as described in any one of claims 1-10, characterized in that, The COPD referred to is mild CPOD, moderate COPD, or severe COPD.

12. The use or method as described in any one of claims 1-11, characterized in that, The COPD is environmental COPD, infection-induced COPD; preferably, the COPD is smoking-related COPD, COPD related to exposure to biological dyes and pollution, COPD with chronic bronchitis, or tuberculosis-related COPD.

Images