Nitric oxide donor-type compound and use thereof

Abstract

The present application relates to a nitric oxide donor-type compound and use thereof in the preparation of a medicament for preventing and / or treating ophthalmic diseases such as glaucoma and ocular hypertension. The compound has a structure represented by formula (I), or is a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of the structure represented by formula (I).

Description

A nitric oxide donor compound and its application Technical Field

[0001] This application belongs to the field of medicinal chemistry research, specifically relating to a nitric oxide donor compound and its application, particularly to a nitric oxide donor compound that inhibits ROCK kinase and / or myosin light chain phosphorylation, and its application in the preparation of drugs for the prevention and / or treatment of ophthalmic diseases such as glaucoma and ocular hypertension. Background Technology

[0002] Glaucoma is an eye disease characterized by intermittent or persistent elevation of intraocular pressure. Sustained high intraocular pressure can damage various tissues of the eye and visual function; without timely treatment, it can lead to complete visual field loss and blindness. Glaucoma is one of the three leading causes of blindness in humans, with an overall incidence of 1% and 2% after age 45. Elevated intraocular pressure is considered a significant risk factor for glaucoma and is currently an interventional factor; therefore, glaucoma treatment to date is based on lowering intraocular pressure.

[0003] ROCK belongs to the serine / threonine protein kinase family. In mammalian cells, ROCK includes two subtypes: ROCKI and ROCKII. Rho is the upstream activator of Rho kinase and a member of the Ras superfamily. Due to its GTPase activity, it is also known as Rho GTPase (such as RhoA, Racl, and Cde42). ROCK receives activation signals from Rho, undergoes phosphorylation at multiple amino acid sites, and mediates a series of downstream phosphorylation / dephosphorylation reactions. ROCK kinase inhibitors can induce myosin light chain dephosphorylation, inhibit transforming growth factor β expression, and form cross-linked actin networks. These changes directly or indirectly affect the morphology, cell contraction / relaxation properties, and intercellular connections of trabecular meshwork cells and Schlemm's tube cells, thereby improving aqueous humor outflow and reducing intraocular pressure.

[0004] Nitric oxide (NO) is an important intercellular signaling agent. It can be synthesized in vivo by nitric oxide synthase or released by drugs such as nitroglycerin and nitroprostene Bunod. In recent years, more and more studies have reported the potential of NO for the treatment of glaucoma. The regulation of NO and its signaling pathway is closely related to aqueous humor dynamics (Invest Ophthalmol Vis Sci, 2014, 55(8):5005-15). Compared with healthy subjects, patients with ocular hypertension have reduced NO production, and exogenous NO administration has been shown to reduce intraocular pressure in these patients (Invest Ophthalmol Vis Sci, 1995, 36(9):1774-84). eNOS knockout mice or animals with impaired GC activity have higher intraocular pressure compared with their wild-type counterparts, further confirming the importance of NO signaling in regulating intraocular pressure (Invest Ophthalmol Vis Sci, 2017, 58(11):4826-4835). Preliminary mechanistic studies have confirmed that NO can reduce aqueous humor outflow resistance through multiple pathways, the most important of which is the diastolic membrane / sac. Some studies have also confirmed that NO can reduce aqueous humor production to some extent.

[0005] Therefore, developing small molecule drugs that simultaneously inhibit ROCK kinase and / or myosin light chain phosphorylation and are also NO donors may achieve better results in reducing intraocular pressure and treating glaucoma than single-function small molecules, which has significant social and economic implications. Summary of the Invention

[0006] In a first aspect, this application provides a compound having the structure shown in Formula (I), or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of the structure shown in Formula (I):

[0007] in:

[0008] R 1 Selected from substituents with a terminal -O-NO2;

[0009] R 2 and R 3 Each is independently selected from hydrogen, deuterium, halogen, hydroxyl, amino, cyano, and C. 1-6 Alkyl groups; for example, H, D, F, Cl, Br, I, OH, NH2, CN, and C. 1-3 alkyl;

[0010] R 4 Selected from hydrogen, deuterium, halogen, hydroxyl, amino, cyano, and optionally surrounded by 1, 2, 3, 4, 5, or 6 R groups. aReplacement C 1-6 Alkyl groups; such as H, D, F, Cl, Br, I, OH, NH2, CN, and optionally surrounded by 1, 2, or 3 R groups. a Replacement C 1-3 alkyl;

[0011] R 5 Selected from hydrogen, deuterium, halogen, hydroxyl, amino, cyano and C 1-6 Alkyl groups; for example, H, D, F, Cl, Br, I, OH, NH2, CN, and C. 1-3 alkyl;

[0012] T 1 Selected from -(CH2) n -, n is selected from 0, 1, and 2;

[0013] T 2 Selected from -(CH2) m -, m is selected from 0, 1, 2 and 3;

[0014] R a Selected from deuterium, halogen, hydroxyl, amino, cyano and C 1-6 Alkyl groups; such as D, F, Cl, Br, I, OH, NH2, CN, and C. 1-3 alkyl.

[0015] In some embodiments, the structural unit Selected from Preferred Other variables T 1 T 2 and R 5 As defined in this application.

[0016] In some implementations, the R 1 It is -Y-ONO2;

[0017] Where Y is selected from C 1-10 Alkylene, C 3-12 Cycloalkylene, 3-12 membered heterocycloalkylene, C 6-10 aryl, 5-10 quinone heteroaryl, C 3-12 Cycloalkyl C 1-6 Alkylene, 3-12 membered heterocyclic alkylene C 1-6 Alkylene, C 6-10 Aromatic C 1-6 Alkylene, 5-10-membered heteroaryl C 1-6 Alkylene, C 1-10 Alkylene (C 1-6 (alkoxide) 1-3 C 3-12 Cycloalkyl C 1-6alkeneoxy, 3-12 membered heterocyclic alkylene C 1-6 alkeneoxy, C 6-10 Aromatic C 1-6 alkeneoxy groups and 5-10 heteroaryl C groups 1-6 Alkyloxy;

[0018] The groups described in Y are independently and arbitrarily selected by m1 R groups. c Replacement, m1 is selected from 0, 1, 2 and 3;

[0019] R c Selected from deuterium, halogen, hydroxyl, amino, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Cyanoalkyl, C 1-6 Carboxyalkyl, C 1-6 Hydroxyalkyl and -(CH2) 0-6 -ONO2; for example, D, F, Cl, Br, I, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Cyanoalkyl, C 1-3 Carboxyalkyl, C 1-3 Hydroxyalkyl and -(CH2) 0-3 -ONO2;

[0020] The 3-12 membered heterocyclic alkyl group, 5-10 membered heteroaryl group, 3-12 membered heterocyclic alkyl group C 1-6 Alkylene, 3-12 membered heterocyclic alkylene C 1-6 alkeneoxy, 5-10 heteroaryl C 1-6 Alkylene and 5-10-membered heteroaryl C 1-6 Each alkoxide independently contains 1, 2, 3 or 4 heteroatoms or heterogroups independently selected from -NH-, -O-, -S- and N.

[0021] In some embodiments, the compound has the structural formula (I-1) as follows:

[0022] in,

[0023] R 2 R 3 R 4 R 5 Y is as defined in this application.

[0024] In some embodiments, the compound has the following structural formula:

[0025] R 2 R 3 R 4 R 5 Y is as defined in this application.

[0026] In some implementations, R in the above general formula 2 and R 3 The variables are independently selected from H, D, F, Cl, Br, I, OH and NH2, and other variables are as defined in this application.

[0027] In some implementations, R in the above general formula 4 The variables are selected from H, D, F, Cl, Br, I, OH, NH2, CN, CH3 and CH2CH3, and other variables are as defined in this application.

[0028] In some implementations, R in the above general formula 5 The variables are selected from H, D, F, Cl, Br, I, OH, NH2, CN and CH3, and other variables are as defined in this application.

[0029] In some implementations, R in the above general formula c The variables are selected from D, F, Cl, Br, I, OH, NH2, CN, CH3, -CH2OH, -ONO2 and -CH2-ONO2, and other variables are as defined in this application.

[0030] In some embodiments, Y in the above general formula is selected from C. 1-5 Alkylene, C 6-10 Alkylene, C 3-8 Monocycloalkyl substructure, C 5-10 Subbridged cycloalkyl, C 6-12 Spirocycloalkyl, 3-8 membered heterocycloalkyl, C 6-8 Alpha-aryl, C 3-8 Monocycloalkyl C 1-4 Alkylene, C 5-10 Subbridged cycloalkyl C 1-4 Alkylene, C 6-12 Spirocycloalkyl C 1-4 Alkylene, 3-8 quinone heterocyclic alkylene C 1-4 Alkylene, C 6-8 Aromatic C 1-4 Alkylene, C 1-6 Alkylene (C 1-4 (alkoxide) 1-3 C 3-8 Monocycloalkyl C 1-3 alkeneoxy, C 5-10 Subbridged cycloalkyl C 1-4 alkeneoxy, C 6-12Spirocycloalkyl C 1-4 alkeneoxy group, 3-8 member heterocyclic alkyl group 14 alkeneoxy and C 6-8 Aromatic C 1-4 Alkyloxy;

[0031] The groups described in Y are independently and arbitrarily selected by m1 R groups. c Replacement, m1 is selected from 0, 1, 2, and 3; R c Selected from D, F, Cl, Br, I, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Cyanoalkyl, C 1-3 Carboxyalkyl, C 1-3 Hydroxyalkyl and -(CH2) 0-3 -ONO2.

[0032] In some embodiments, the Y in the above general formula is selected from -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)8-, and -(CH2) 10 -, -CH(CH3)-, -CH2-(OCH2CH2)3-, Other variables are as defined in this application.

[0033] In some embodiments, Y in the above general formula is selected from... Other variables are as defined in this application.

[0034] In some embodiments, the Y in the above general formula is selected from CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)8-, -(CH2) 10 -, -CH(CH3)-, -CH(CH3)CH2-, -C(CH3)2-, -C(CH3)2CH2-, -C(CH3)2(CH2)3-, -CH2-(OCH2CH2)3-, Other variables are as defined in this application.

[0035] In some embodiments, Y in the above general formula is selected from... Other variables are as defined in this application.

[0036] In some embodiments, the compound is selected from the following structural formulas:

[0037] L 1 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 R 5 R c m1 is as defined in this application.

[0038] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0039] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0040] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0041] R c Selected from deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0042] L 1 Selected from single bonds, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene;

[0043] m1 is either 0 or 1.

[0044] In some embodiments, the compound is selected from the following structural formulas:

[0045] L 1 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 R 5 R c m1 is as defined in this application.

[0046] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0047] R 4Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0048] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0049] R c Selected from deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0050] L 1 Selected from single bonds, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene;

[0051] m1 is either 0 or 1.

[0052] In some embodiments, the compound is selected from the following structural formulas:

[0053] L 2 Selected from single bond, C 1-5 Alkylene, C 6-10 Alkylene, with one or two -(CH2) groups 0-3 -ONO2 replaces C 1-6 Alkylene, R 2 R 3 R 4 and R 5 As defined in this application,

[0054] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0055] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0056] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0057] L 2 Selected from single bond, C 1-6 Alkylene, C 7-10 Alkylene, with one or two -(CH2) groups 0-3 -ONO2 replaces C 1-6 Alkyl groups, such as -CH2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)8-, -(CH2) 10-、-C(CH3)2-、

[0058] In some embodiments, the compound is selected from the following structural formulas:

[0059] L 2 Selected from single bond, C 1-5 Alkylene, C 6-10 Alkylene, with one or two -(CH2) groups 0-3 -ONO2 replaces C 1-6 Alkylene, R 2 R 3 R 4 and R 5 As defined in this application,

[0060] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0061] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0062] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0063] L 2 Selected from single bond, C 1-6 Alkylene, C 7-10 Alkylene, with one or two -(CH2) groups 0-3 -ONO2 replaces C 1-6 Alkyl groups, such as -CH2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)8-, -(CH2) 10 -、-C(CH3)2-、

[0064] In some embodiments, the compound is selected from the following structural formulas:

[0065] L 3 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 R 5 Rc m1 is as defined in this application.

[0066] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0067] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0068] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0069] R c Selected from deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0070] L 3 Selected from single bonds, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene;

[0071] m1 is either 0 or 1.

[0072] In some embodiments, the compound is selected from the following structural formulas:

[0073] L 3 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 R 5 R c m1 is as defined in this application.

[0074] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0075] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0076] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0077] R c Selected from deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl;

[0078] L 3 Selected from single bonds, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene;

[0079] m1 is either 0 or 1.

[0080] In some embodiments, the compound is selected from the following structural formulas:

[0081] L 4 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 R 5 R c m1 is as defined in this application.

[0082] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0083] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0084] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0085] R c Selected from deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and -(CH2). 0-3 -ONO2;

[0086] L 4 Selected from single bonds, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene;

[0087] m1 is either 0 or 1.

[0088] In some embodiments, the compound is selected from the following structural formulas:

[0089] L 4 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 R 5 R c m1 is as defined in this application.

[0090] Preferably, R 2and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0091] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0092] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0093] R c Selected from deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and -(CH2). 0-3 -ONO2;

[0094] L 4 Selected from single bonds, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene;

[0095] m1 is either 0 or 1.

[0096] In some embodiments, the compound is selected from the following structural formulas:

[0097] L 5 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 and R 5 As defined in this application,

[0098] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0099] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0100] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0101] L 5 It is selected from single bond, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene.

[0102] In some embodiments, the compound is selected from the following structural formulas:

[0103] L 5 Selected from single bond, C 1-6 Alkylene, R 2 R 3 R 4 and R 5 As defined in this application,

[0104] Preferably, R 2 and R 3 Each of the following is independently selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0105] R 4 Selected from hydrogen, deuterium, halogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0106] R 5 Selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl;

[0107] L 5 It is selected from single bond, methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, or tert-butylene.

[0108] Secondly, this application provides a compound having the structure shown below, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

[0109] The compounds provided in this application have the structures shown below, or their stereoisomers, tautomers, solvates, metabolites, pharmaceutically acceptable salts, or prodrugs:

[0110] In some embodiments, the compound has the structure shown below, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

[0111] In some embodiments, the compound has the structure shown below, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

[0112] In some implementations, the pharmaceutically acceptable salt is selected from hydrochloride salts.

[0113] Thirdly, this application provides a pharmaceutical composition comprising the compound described in the first or second aspect, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, medium, or combination thereof.

[0114] In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the compound described in the first or second aspect as an active ingredient, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, medium, or combination thereof.

[0115] Fourthly, this application provides the use of the compounds described in the first aspect, the second aspect, or the compositions described in the third aspect in the preparation of medicaments that inhibit ROCK kinase and / or myosin light chain phosphorylation.

[0116] In some embodiments, the drug that inhibits ROCK kinase and / or myosin light chain phosphorylation is a drug for treating glaucoma and / or ocular hypertension.

[0117] Fifthly, this application provides a method for inhibiting ROCK kinase and / or myosin light chain phosphorylation, comprising contacting the cells with or administering to the subject the compound of the first aspect, the second aspect, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug, or a pharmaceutical composition as described in the third aspect.

[0118] In some implementations, the subject is a mammal, preferably a human.

[0119] In a sixth aspect, this application provides a method for treating glaucoma and / or ocular hypertension in a patient, comprising administering to the patient the compound described in the first aspect, the second aspect, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug, or the pharmaceutical composition described in the third aspect.

[0120] In some embodiments, the glaucoma and / or ocular hypertension are associated with increased activity of ROCK kinase and / or myosin light chain phosphorylation.

[0121] Compared with the prior art, this application has the following beneficial effects:

[0122] The metabolites of the compounds provided in this application not only inhibit the phosphorylation of ROCK kinase and / or myosin light chain, but also release nitric oxide. They have better effects on reducing intraocular pressure and treating glaucoma than single-function small molecules, and have great clinical application prospects. Detailed Implementation

[0123] Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient. The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0124] The term "pharmaceutically acceptable salt" refers to the salt of the compounds in this application, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this application, with a specific substituent. When the compounds in this application contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds in this application contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; as well as salts of amino acids (such as arginine) and salts of organic acids such as glucuronic acid. Certain specific compounds of this application contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0125] The pharmaceutically acceptable salts of this application can be synthesized from parent compounds containing an acid radical or a base using conventional chemical methods. Generally, such salts are prepared by reacting these compounds, in their free acid or base form, with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of both.

[0126] In addition to the salt form, the compounds provided in this application also exist in prodrug form. The prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to be converted into the compounds of this application. Furthermore, the prodrugs can be converted into the compounds of this application in the in vivo environment via chemical or biochemical methods.

[0127] Some compounds of this application may exist in non-solventized or solvated forms, including hydrated forms. Generally, solvated and non-solventized forms are equivalent and are both included within the scope of this application.

[0128] The compounds of this application may exist in specific geometric or stereoisomeric forms. This application envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this application. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this application.

[0129] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0130] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0131] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0132] Unless otherwise stated, "(D)" or "(+)" indicates right-handed rotation, "(L)" or "(-)" indicates left-handed rotation, and "(DL)" or "(±)" indicates racemic rotation.

[0133] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and straight dashed key

[0134] The compounds described in this application may exist in specific forms. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. If tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomer can be achieved. For example, proton tautomers (also called prototropic tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions involving the rearrangement of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between the two tautomers, pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0135] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0136] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0137] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of this application, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide a pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to carbamates). The compounds of this application may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, compounds can be labeled with radioactive isotopes, such as tritium ( 3 H), Iodine-125 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds in this application, regardless of radioactivity, are included within the scope of this application.

[0138] The terms “optional” or “optionally” refer to events or conditions that may occur but are not required to occur as described below, and the description includes both cases where said events or conditions occur and cases where said events or conditions do not occur.

[0139] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents can be arbitrary on a chemically feasible basis.

[0140] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0141] Unless otherwise specified, the term "C" 1-10 "alkylene" is used to denote a straight-chain or branched saturated divalent hydrocarbon group consisting of 1 to 10 carbon atoms. The C 1-10 Alkylenes include C 1-9 C 1-8 C 1-6 C 1-5 C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C 10 C8, C7, C6, and C5 alkylene groups, etc. 1-10 Examples of alkyl groups include, but are not limited to, methylene, ethylene, propylene, butylene, etc.

[0142] Unless otherwise specified, the term "C" 1-6 "Alkoxy" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to the rest of the molecule by an oxygen atom. The C 1-3 Alkoxy groups include C 1-2 C 2-3 C3 alkoxy groups, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

[0143] Unless otherwise specified, the term "C" 3-12 "Cycloalkylene" refers to a saturated cyclic divalent hydrocarbon group consisting of 3 to 12 carbon atoms, which can be monocyclic or bicyclic systems. Bicyclic systems include spirocyclic, fused, and bridged rings. 3-12 Cycloalkylene compounds include C 5-12 C 3-8 and C 4-9 Cycloalkylene compounds, etc. C 3-12 Examples of cycloalkylene compounds include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, spiro[3.5]nonane, etc.

[0144] Unless otherwise specified, the term "3-12 membered heterocyclic alkylene" on its own or in combination with other terms refers to a saturated cyclic divalent group consisting of 3 to 12 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). p (where p is 1 or 2). It includes monocyclic and bicyclic systems, wherein bicyclic systems include spirocyclic, fused, and bridged rings. Furthermore, with respect to the “3-8 membered heterocyclic alkylene,” the heteroatom can occupy the connection position between the heterocyclic alkylene and the rest of the molecule. The 3-12 membered heterocyclic alkylene includes 3-10, 3-8, 4-12, 5-12, 4, 5, and 10 membered heterocyclic alkylene, etc. Examples of 3-12 heterocyclic alkylene include, but are not limited to, ethylene oxide, aziridinyl, aziridinyl, oxobutyl, tetrahydrofuranyl, dioxacyclopentenyl, pyrrolidine, pyrrolidone, imidazoalkyl, pyrazolidine, pyrrololinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazine, or trithiaalkylene.

[0145] Unless otherwise specified, the term "C" 6-10 "Aryl" refers to a monocyclic or fused polycyclic aromatic divalent group composed of 6 to 10 carbon atoms with a conjugated π-electron system. 6-10 Examples of arylene groups include, but are not limited to, phenylene or naphthylene.

[0146] Unless otherwise specified, the term "5-10 heteroaryl" refers to a cyclic divalent group consisting of 5 to 10 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. It can be a monocyclic, fused bicyclic, or fused tricyclic system, wherein each ring is aromatic. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). p (where p is 1 or 2). A 5-10 membered heteroaryl group can be attached to the rest of the molecule via a heteroatom or carbon atom. The 5-10 membered heteroaryl groups include 5-8, 5-7, 5-6, 5, and 6 membered heteroaryl groups. Examples of the 5-10 membered heteroaryl groups include, but are not limited to, thiopheneyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl, pyridinyl, pyridinyl, or triazinyl.

[0147] Unless otherwise specified, the term "C" 1-6"Haloalkyl" means that at least one hydrogen atom in a saturated hydrocarbon group consisting of 1 to 6 carbon atoms, representing a straight chain or branched chain, is replaced by a halogen atom such as F, Cl, Br, or I. The C 1-3 Haloalkyl groups include C1, C2, and C3 haloalkyl groups, etc. 1-3 Examples of haloalkyl groups include, but are not limited to, -CH2Cl, -CH2CH2Cl, -CH2CH2CH2Cl, etc.

[0148] Unless otherwise specified, the term "C" 1-6 "Cyanoalkyl" means that at least one hydrogen atom in a saturated hydrocarbon group consisting of 1 to 6 carbon atoms, representing a straight chain or branched chain, is replaced by a cyano group. The C 1-3 Cyanoalkyl groups include C1, C2, and C3 cyanoalkyl groups, etc. 1-3 Examples of cyanoalkyl groups include, but are not limited to, -CH2CN, -CH2CH2CN, -CH2CH2CH2CN, etc.

[0149] Unless otherwise specified, the term "C" 1-6 "Carboxyalkyl" means that at least one hydrogen atom in a saturated hydrocarbon group consisting of 1 to 6 carbon atoms, representing a straight chain or branched chain, is replaced by a carboxyl group. 1-3 Carboxylalkyl groups include C1, C2, and C3 carboxylalkyl groups, etc. 1-3 Examples of cyanoalkyl groups include, but are not limited to, -CH2COOH, -CH2CH2COOH, -CH2CH2CH2COOH, etc.

[0150] Unless otherwise specified, the term "C" 1-6 "Hydroxyalkyl" means that at least one hydrogen atom in a saturated hydrocarbon group consisting of 1 to 6 carbon atoms, representing a straight chain or branched chain, is replaced by a hydroxyl group. 1-3 Hydroxyalkyl groups include C1, C2, and C3 hydroxyalkyl groups, etc. 1-3 Examples of cyanoalkyl groups include, but are not limited to, -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, etc.

[0151] Unless otherwise specified, when multiple groups are used in combination in this application, such as "C 3-12 Cycloalkyl C 1-6 "alkylene" indicates C 3-12 Cycloalkylene and C 1-6 Alkyl groups are linked. When the listed substituents do not specify which atom they are attached to the substituted group, they can be bonded to any of their atoms. For example, a cyclopentyl group can be attached to the substituted group via any carbon atom on the cyclopentyl ring. When the linked group is not specified in terms of its direction of attachment, the direction is arbitrary. For example, in -C(=O)-Y-ONO2, the linking group Y is C. 3-12Cycloalkyl C 1-6 Alkylene, where Y can be connected to -C(=O)- and ONO2 in the same direction as the left-to-right reading order to form -C(=O)-C 3-12 Cycloalkyl C 1-6 Alkylene-ONO2 can also be formed by connecting -C(=O)- and ONO2 in the reverse order of reading from left to right to form -C(=O)-C. 1-6 Alkylene C 3-12 Cycloalkylene-ONO2. Combinations of the linking group, substituents, and / or their variants are permitted only if such combinations produce a stable compound. When the number of a linking group is 0, such as -(CR)0-, it indicates that the linking group is a single bond.

[0152] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-10 Including C1, C2, C3, C4, C5, C6, C7, C8, C9 and C 10 It also includes any range from n to n+m, such as C 1-0 Including C 1-3 C 1-6 C 1-9 C 3-6 C 3-9 C 3-10 C 6-9 C 6-10 and C 7-10 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-8-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, and 8-membered rings, and also include any range from n to n+m. For example, 3-8-membered rings include 3-6-membered, 3-7-membered, 5-6-membered, 5-7-membered, 6-7-membered, and 6-8-membered rings, etc.

[0153] When applicable for treatment, a therapeutically effective amount of the compound of this application may be administered as an unprocessed chemical drug or as an active ingredient in a pharmaceutical composition. Therefore, this application also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of this application and one or more pharmaceutically acceptable carriers, diluents, or excipients. The term "therapeuticly effective amount" as used herein refers to the total amount of each active component sufficient to demonstrate a meaningful patient benefit (e.g., reduction of viral load). When administered alone using a single active ingredient, the term refers only to that component. When used in combination, the term refers to the combined amount of active ingredients that, regardless of combination, sequential or simultaneous administration, produce a therapeutic effect. The carrier, diluent, or excipient must be acceptable in the sense of compatibility with other components of the formulation and harmlessness to the recipient. According to another aspect of this application, a method for preparing a pharmaceutical formulation is also provided, comprising mixing the compound of this application with one or more pharmaceutically acceptable carriers, diluents, or excipients. As used in this application, the term "pharmaceutical acceptable" means that the compounds, raw materials, compositions, and / or dosage forms of this application are suitable for contact with patient tissues without excessive toxicity, irritation, allergic reactions, or other problems and complications commensurate with a reasonable benefit / risk ratio, and are effective for their intended use, within the limits of reasonable medical judgment.

[0154] It should be understood that, in addition to the ingredients specifically mentioned above, the formulation may also include other ingredients commonly used in the art in relation to the type of formulation, such as flavoring agents, for example, such formulations suitable for oral administration may include flavoring agents.

[0155] The “effective amount” or “effective dose” of the compound or pharmaceutically acceptable composition of this application refers to an effective amount for treating or reducing the severity of one or more of the conditions mentioned in this application. According to the methods of this application, the compound and its composition can be administered at any dosage and via any route of administration to effectively treat or reduce the severity of the disease. The precise amount required will vary depending on the patient’s condition, which may depend on age, the patient’s general condition, the severity of the infection, specific factors, route of administration, etc. The compound or composition of this application may be administered in combination with one or more other therapeutic agents, as discussed in this application.

[0156] Generally, the compounds of this application can be prepared by the methods described herein. The following reaction schemes and examples are used to further illustrate the content of this application.

[0157] Those skilled in the art will recognize that the chemical reactions described in this application can be suitably used to prepare many other compounds of this application, and that other methods for preparing the compounds of this application are considered to be within the scope of this application. For example, the synthesis of those non-illustrative compounds according to this application can be successfully accomplished by those skilled in the art through modification methods, such as appropriately protecting interfering groups, utilizing other known reagents besides those described in this application, or making some conventional modifications to the reaction conditions. Furthermore, the reactions disclosed in this application or the known reaction conditions are also generally accepted to be applicable to the preparation of other compounds of this application.

[0158] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with embodiments. The specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of this application. Furthermore, descriptions of well-known structures and technologies are omitted in the following description to avoid unnecessarily obscuring the concepts of this disclosure. Such structures and technologies have also been described in numerous publications.

[0159] [Amended according to Rule 26, 31.07.2025] Example 1: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethyl)benzoate (Compound 1)

[0160] Step 1: Preparation of 4-(2-(nitoxy)ethyl)benzoic acid (compound 1b)

[0161] Under nitrogen protection, silver nitrate (0.44 g, 2.62 mmol) was added to a solution of 4-(2-bromoethyl)benzoic acid (compound 1a) (0.5 g, 2.2 mmol) in acetonitrile (10 mL). The resulting mixture was heated to reflux and stirred for 16 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (50 mL) and filtered. The filtrate was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 4-(2-(nitoxy)ethyl)benzoic acid (compound 1b) (0.4 g, 87% yield, white solid). 1 HNMR(500MHz,DMSO-d6)δ:12.90(s,1H),7.90(d,2H),7.43(d,2H),4.79(t,2H),3.10(t,2H)ppm. LC-MS m / z(ESI)=210.1[MH] - .

[0162] Step 2: Preparation of chloromethyl 4-(2-(nitoxy)ethyl)benzoate (compound 1c)

[0163] Under nitrogen protection, 4-(2-(nitoxy)ethyl)benzoic acid (compound 1b) (0.4 g, 1.9 mmol), dichloromethane (10 mL), water (10 mL), sodium bicarbonate (0.4 g, 4.7 mmol), and tetrabutylammonium fluoride (0.4 mL, 0.4 mmol, 1N) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (0.34 g, 2.27 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain chloromethyl 4-(2-(nitoxy)ethyl)benzoate (compound 1c) (0.35 g, 71% yield, colorless oil).

[0164] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethyl)benzoate (compound 1d)

[0165] Under nitrogen protection, sodium hydride (34.4 mg, 0.9 mmol, 60%) was added to a solution of (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate (compound M1, synthetic method referred to patent WO2020253882A1) (0.25 g, 0.61 mmol) in N,N-dimethylformamide (5 mL) at 0 °C. The mixture was stirred for 20 min, and then 4-(2-(nitoxy)ethyl)benzoate (compound 1c) (0.23 g, 0.86 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 0.5 h. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched by adding water (10 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 2), washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethyl)benzoate (compound 1d) (0.35 g, 90% yield). LC-MS m / z (ESI) = 631.6 [M+H] + .

[0166] Step 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethyl)benzoate (Compound 1)

[0167] A solution of dioxane hydrochloride (0.6 mL, 2.4 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl4-(2-(nitrooxy)ethyl)benzoate (compound 1d) (0.15 g, 0.24 mmol) in ethyl acetate (5 mL). The resulting mixture was stirred at room temperature for 6 hours. The mixture was concentrated under reduced pressure, and the residue was separated by high pressure to give compound (S)-(5-((3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl4-(2-(nitrooxy)ethyl)benzoate (compound 1) (64 mg, 47% yield, white solid). 1 H NMR(500MHz, CDCl3)δ:8.81(d,1H),8.38(d,1H),8.05(d,2H),7.54(t,1H),7.38-7.29(m,3H),6.20(s,2H),4.67(t,2H) ,3.87(dd,1H),3.76-3.60(m,3H),3.25(dd,1H),3.10(t,2H),2.75(s,3H),1.87(dt,1H),1.60(s,2H),1.28(d,1H)ppm. LC-MS m / z(ESI)=531.2[M+H] + .

[0168] [Amended according to Rule 26, 31.07.2025] Example 2: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (Compound 2)

[0169] Step 1: Preparation of 6-(nitrooxy)hexanoic acid (compound 2b)

[0170] Under nitrogen protection, silver nitrate (1.1 g, 6.5 mmol) was added to a solution of 6-bromohexanoic acid (compound 2a) (1.0 g, 5.1 mmol) in acetonitrile (10 mL). The resulting mixture was heated to reflux and stirred for 16 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (80 mL) and filtered. The filtrate was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 6-(nitrooxy)hexanoic acid (compound 2b) (0.8 g, 88% yield, colorless oil). LC-MS m / z (ESI) = 176.0 [MH] - .

[0171] Step 2: Preparation of chloromethyl 6-(nitrooxy)hexanoate (compound 2c)

[0172] Under nitrogen protection, 6-(nitrooxy)hexanoic acid (compound 2b) (0.8 g, 4.5 mmol), dichloromethane (15 mL), water (15 mL), sodium bicarbonate (0.9 g, 10.7 mmol), and tetrabutylammonium fluoride (0.9 mL, 0.9 mmol, 1N) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (0.9 g, 6.0 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain chloromethyl 6-(nitrooxy)hexanoate (compound 2c) (0.65 g, yield 64%, colorless oil).

[0173] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 2d)

[0174] Under nitrogen protection, sodium hydride (38.0 mg, 1.0 mmol, 60%) was added to a solution of (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate (compound M1) (0.3 g, 0.7 mmol) in N,N-dimethylformamide (5 mL) at 0 °C. The mixture was stirred for 20 min, and methyl 6-(nitrooxy)hexanoate (2c) (0.23 g, 1.0 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 h. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched by adding water (10 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 2), washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 2d) (0.3 g, yield 68%, white solid). LC-MS m / z (ESI) = 597.4 [M+H] + .

[0175] Step 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 2)

[0176] A solution of dioxane hydrochloride (1 mL, 4.0 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 2d) (0.3 g, 0.5 mmol) in ethyl acetate (5 mL). The resulting mixture was stirred at room temperature for 8 hours. The mixture was concentrated under reduced pressure, and the residue was separated by high pressure to give methyl (S)-(5-(((3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 2) (40 mg, 16% yield, white solid). 1HNMR(500MHz, CDCl3)δ:8.76(d,1H),8.35(d,1H),7.52(t,1H),7.20(s,1H),5.92(s,2H),4.41(t,2H),3.89-3.82(m,1H),3.66(m ,3H),3.23(dd,1H),2.71(s,3H),2.39(t,2H),2.28-2.19(m,1H),1.86(dt,1H),1.76-1.65(m,4H),1.42(dt,2H),1.26(d,2H)ppm. LC-MS m / z(ESI)=497.4[M+H] + .

[0177] [Amended according to Rule 26, 31.07.2025] Example 3: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)5-(nitrooxy)valerate (compound 3)

[0178] Step 1: Preparation of 5-(nitrooxy)valerate (compound 3b)

[0179] Under nitrogen protection, silver nitrate (1.1 g, 6.5 mmol) was added to a solution of 5-bromopentanoic acid (compound 3a) (1.0 g, 5.5 mmol) in acetonitrile (10 mL). The resulting mixture was heated to reflux and stirred for 16 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (80 mL) and filtered. The filtrate was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 5-(nitrooxy)pentanoic acid (compound 3b) (0.9 g, 99% yield, colorless oil). LC-MS m / z (ESI) = 162.0 [MH] - .

[0180] Step 2: Preparation of chloromethyl 5-(nitrooxy)valerate (compound 3c)

[0181] Under nitrogen protection, 5-(nitrooxy)valerate (compound 3b) (0.9 g, 5.5 mmol), dichloromethane (15 mL), water (15 mL), sodium bicarbonate (1.2 g, 14.3 mmol), and tetrabutylammonium fluoride (1.1 mL, 1.1 mmol, 1N) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.1 g, 7.4 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain chloromethyl 5-(nitrooxy)valerate (compound 3c) (0.5 g, yield 43%, colorless oil).

[0182] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)5-(nitrooxy)valerate (compound 3d)

[0183] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (0.3 g, 0.7 mmol), tetrahydrofuran (5 mL), methyl 5-(nitrooxy)valerate (compound 3c) (0.15 g, 0.7 mmol) and cesium carbonate (0.23 g, 0.7 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)5-(nitrooxy)valerate (compound 3d) (0.48 g, yield 68%, white solid). LC-MS m / z (ESI) = 583.5 [M+H] + .

[0184] Step 4: Preparation of methyl (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)5-(nitrooxy)valerate (compound 3)

[0185] A solution of dioxane hydrochloride (1 mL, 4.0 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)5-(nitrooxy)valerate (compound 3d) (0.3 g, 0.5 mmol) in ethyl acetate (5 mL). The resulting mixture was stirred at room temperature for 8 hours. The mixture was concentrated under reduced pressure, and the residue was separated by high pressure to give methyl (S)-(5-((3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)5-(nitrooxy)valerate (compound 3) (40 mg, 16% yield, white solid). 1 H NMR(500MHz, CDCl3)δ:8.76(d,1H),8.35(d,1H),7.52(t,1H),7.19(s,1H),5.93(s,2H),4.48-4.38(m,2H),3.90-3.84(m,1H),3.7 0-3.63(m,3H),3.23(dd,1H),2.71(s,3H),2.42(t,2H),2.27-2.19(m,1H),1.89-1.82(m,1H),1.78-1.73(m,4H),1.26(d,2H)ppm. LC-MS m / z(ESI)=483.5[M+H] + .

[0186] [Amended according to Rule 26, 31.07.2025] Example 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-4-(2-(nitrooxy)ethyl)benzoate (Compound 4)

[0187] Step 1: Preparation of methyl 2-methyl-4-vinylbenzoate (compound 4b)

[0188] Under nitrogen protection, potassium vinyltrifluoroborate (4.2 g, 31.4 mmol), palladium dichloride bis(triphenylphosphine) chloride (0.92 g, 1.31 mmol), and cesium carbonate (17.0 g, 52.2 mmol) were sequentially added to a tetrahydrofuran (120 mL) and water (12 mL) solution of methyl 4-bromo-2-methylbenzoate (compound 4a) (6.0 g, 26.2 mmol). The resulting mixture was heated to reflux and stirred for 8 hours. The mixture was filtered, extracted with ethyl acetate (200 mL), and the combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound methyl 2-methyl-4-vinylbenzoate (compound 4b) (4.0 g, 87% yield, colorless oil). 1 H NMR (400MHz, CDCl3) δ: 7.93 (d, 1H), 7.36-7.26 (m, 2H), 6.73 (dd, 1H), 5.87 (d, 1H), 5.38 (d, 1H), 3.92 (s, 3H), 2.64 (s, 3H) ppm. LC-MS m / z(ESI)=177.2[M+H] + .

[0189] Step 2: Preparation of methyl 4-(2-hydroxyethyl)-2-methylbenzoate (compound 4c)

[0190] Under nitrogen protection at 0°C, a borane tetrahydrofuran solution (34.0 mL, 34.0 mmol, 1N) was added dropwise to a tetrahydrofuran solution (100 mL) of methyl 2-methyl-4-vinylbenzoate (compound 4b) (4.0 g, 22.7 mmol). The resulting mixture was heated to room temperature and stirred for 1 hour. The mixture was then cooled to 0°C, and sodium hydroxide aqueous solution (1.8 g, 45.4 mmol, 2N) and hydrogen peroxide (7.7 g, 68.1 mmol, 30%) were added dropwise sequentially. After the addition was complete, the mixture was kept at room temperature for 30 min, then cooled to room temperature and stirred for 1 hour. The reaction was quenched by adding saturated sodium bisulfite solution (100 mL) at 0°C. The pH was adjusted to 2 with hydrochloric acid aqueous solution, and the mixture was extracted with ethyl acetate (200 mL × 2). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated under medium pressure to obtain methyl 4-(2-hydroxyethyl)-2-methylbenzoate (compound 4c) (2.4 g, yield 55%, colorless oil). 1H NMR (400MHz, CDCl3) δ: 7.88(d,1H),7.12(d,2H),3.89(s,3H),3.87(d,2H),2.87(t,2H),2.60(s,3H),1.89(s,1H)ppm. LC-MS m / z(ESI)=195.2[M+H] + .

[0191] Step 3: Preparation of 4-(2-hydroxyethyl)-2-methylbenzoic acid (compound 4d)

[0192] Under nitrogen protection, an aqueous solution of sodium hydroxide (0.74 g, 18.5 mmol, 2N) was added to a methanol (30 mL) solution of methyl 4-(2-hydroxyethyl)-2-methylbenzoate (compound 4c) (2.4 g, 12.4 mmol), and the mixture was stirred at 50 °C for 4 hours. The reaction was monitored by TLC until complete. The mixture was cooled to room temperature, the pH was adjusted to 2 with aqueous hydrochloric acid, and the mixture was extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 4-(2-hydroxyethyl)-2-methylbenzoic acid (compound 4d) (1.9 g, 85% yield, white solid). 1 H NMR(400MHz, DMSO-d6)δ:12.59(s,1H),7.76(d,1H),7.14(d,2H),4.68(s,1H),3.63(t,2H),2.74(t,2H),2.51(s,3H)ppm. LC-MS m / z(ESI)=181.2[M+H] + .

[0193] Step 4: Preparation of 2-methyl-4-(2-(nitoxy)ethyl)benzoic acid (compound 4e)

[0194] Under nitrogen protection, concentrated nitric acid (12 mL) and concentrated sulfuric acid (2.8 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 20 min to prepare a mixed acid solution. A solution of 4-(2-hydroxyethyl)-2-methylbenzoic acid (compound 4d) (1.1 g, 6.1 mmol) in dichloromethane (40 mL) was added dropwise, and the mixture was brought back to room temperature and stirred for 5 hours. The reaction was monitored by TLC until complete. The mixture was cooled to 0 °C, and the reaction was quenched with water (50 mL). The mixture was separated, extracted with ethyl acetate (100 mL × 3), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain compound 2-methyl-4-(2-(nitoxy)ethyl)benzoic acid (compound 4e) (1.4 g, yield 66%, white solid). LC-MS m / z (ESI) = 226.2 [M+H] + .

[0195] Step 5: Preparation of chloromethyl 2-methyl-4-(2-(nitoxy)ethyl)benzoate (compound 4f)

[0196] Under nitrogen protection, 2-methyl-4-(2-(nitoxy)ethyl)benzoic acid (compound 4e) (1.4 g, 6.2 mmol), dichloromethane (40 mL), water (40 mL), sodium bicarbonate (2.6 g, 31.1 mmol), and tetrabutylammonium bisulfate (0.4 g, 1.2 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.5 g, 10.1 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparative separation to obtain chloromethyl 2-methyl-4-(2-(nitoxy)ethyl)benzoate (compound 4f) (1.3 g, yield 76%, colorless oil). 1 H NMR (500MHz, CDCl3) δ: 8.00-7.94(m,1H),7.16(s,2H),5.96(s,2H),4.68(t,2H),3.07(t,2H),2.65(s,3H)ppm.

[0197] Step 6: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-4-(2-(nitrooxy)ethyl)benzoate (compound 4g)

[0198] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (450 mg, 1.1 mmol), methyltetrahydrofuran (15 mL), methyl 2-methyl-4-(2-(nitoxy)ethyl)benzoate chloromethyl ester (compound 4f) (600 mg, 2.2 mmol), and cesium carbonate (521 mg, 1.6 mmol) were added sequentially to the reaction flask. The mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was then filtered, dried over anhydrous sodium sulfate, and filtered again. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give methyl (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-4-(2-(nitrooxy)ethyl)benzoate (4 g of compound) (0.54 g, yield 76%, white solid). LC-MS m / z (ESI) = 645.2 [M+H] + .

[0199] Step 7: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-4-(2-(nitrooxy)ethyl)benzoate (compound 4)

[0200] A solution of dioxane hydrochloride (0.38 mL, 1.5 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-4-(2-(nitrooxy)ethyl)benzoate (compound 4 g) (0.1 g, 0.15 mmol) in ethyl acetate (5 mL). The resulting mixture was stirred at room temperature for 8 hours. The mixture was concentrated under reduced pressure, and the residue was separated by high pressure to give compound (S)-(5-(((3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-4-(2-(nitrooxy)ethyl)benzoate (compound 4) (55 mg, 65% yield, white solid). 1H NMR(400MHz, CDCl3)δ:8.78(dd,1H),8.35(dd,1H),7.94(d,1H),7.52(t,1H),7.32(s,1H),7.08(d,2H),6.14(s,2H),4.62(t,2H),3.89-3. 82(m,1H),3.72-3.60(m,3H),3.24(dd,1H),3.01(t,2H),2.73(s,3H),2.60(s,3H),2.23(m,1H),1.91-1.80(m,1H),1.36-1.24(m,2H)ppm. LC-MS m / z(ESI)=545.2[M+H] + .

[0201] [Amended according to Rule 26, 31.07.2025] Example 5: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-((nitrooxy)methyl)1-cyclohexane hydrochloride (compound 5)

[0202] Step 1: Preparation of (1R,4R)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid (compound 5b)

[0203] Under nitrogen protection, concentrated nitric acid (6 mL) and concentrated sulfuric acid (6 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 60 min to prepare a mixed acid solution. A solution of (1R,4R)-4-(hydroxymethyl)cyclohexane-1-carboxylic acid (compound 5a) (1.5 g, 9.5 mmol) in dichloromethane (100 mL) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (50 mL). The mixture was separated, extracted with dichloromethane (100 mL × 3), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain compound (1R,4R)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid (compound 5b) (1.5 g, 78% yield, white solid). 1 H NMR(500MHz, CDCl3)δ:4.28(d,2H),2.30(m,1H),2.15-2.04(m,2H),1.96-1.86(m,2H),1.84-1.70(m,1H),1.47(m,2H),1.10(m,2H)ppm.

[0204] Step 2: Preparation of (1R,4R)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid chloromethyl ester (compound 5c)

[0205] Under nitrogen protection, (1R,4R)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid (compound 5b) (1.0 g, 4.9 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (2.0 g, 23.8 mmol), and tetrabutylammonium bisulfate (0.17 g, 0.5 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethylsulfonyl chloride (1.1 g, 7.4 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain compound (1R,4R)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid chloromethyl ester (compound 5c) (1.1 g, yield 89%, colorless oil).

[0206] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 5d)

[0207] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)tert-butyl carbamate (compound M1) (0.5 g, 1.23 mmol), methyltetrahydrofuran (20 mL), (1R,4R)-4-((nitoxy)methyl)cyclohexane-1-carboxylate (compound 5c) (0.4 g, 1.6 mmol), and cesium carbonate (0.45 g, 1.4 mmol) were added sequentially to the reaction flask. The mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was then filtered, dried over anhydrous sodium sulfate, and filtered again. The filtrate was concentrated under reduced pressure, and the residue was separated under medium pressure to obtain compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 5d) (0.6 g, yield 79%, white solid). 1H NMR(500MHz, CDCl3)δ:8.78(dd,1H),8.06(dd,1H),7.52(t,1H),7.19(s,1H),5.91( s,2H),4.79(s,1H),4.36(s,1H),4.25(d,2H),3.67(dt,2H),3.52(s,1H),3.41(d,1 H),2.69(s,3H),2.37-2.28(m,2H),2.07(d,1H),2.03-1.98(m,1H),1.91-1.84(m,2 H),1.74(m,1H),1.46(s,9H),1.44-1.40(m,1H),1.28-1.22(m,2H),1.05(m,2H)ppm. LC-MS m / z(ESI)=623.6[M+H] + .

[0208] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-((nitrooxy)methyl)1-cyclohexane hydrochloride (compound 5)

[0209] A solution of dioxane hydrochloride (1.6 mL, 6.4 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)6-(nitrooxy)hexanoate (compound 5d) (0.4 g, 0.64 mmol) in 10 mL of ethyl acetate at room temperature. The resulting mixture was stirred at room temperature for 8 hours. The mixture was filtered and dried to give compound (5-(((S)-3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-((nitrooxy)methyl)1-cyclohexane hydrochloride (compound 5) (290 mg, 81% yield, white solid). 1H NMR(400MHz, DMSO-d6)δ:8.67-8.57(m,4H),8.20(dd,1H),7.70(t,1H),7.56(s,1H),5.90(s,2H),4.35(d,2H),4.04(d,1H),3.81-3.68(m, 2H),3.58-3.49(m,2H),2.59(s,3H),2.44(dt,1H),2.36-2.17(m,2H),1.95-1.87(m,2H),1.70(m,3H),1.34(m,2H),1.12-1.00(m,2H)ppm. LC-MS m / z(ESI)=523.6[M+H] + .

[0210] [Amended according to Rule 26, 31.07.2025] Example 6: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-3-(nitrooxy)propionate hydrochloride (compound 6)

[0211] Step 1: Preparation of methyl 2,2-dimethyl-3-(nitoxy)propionate (compound 6b)

[0212] Under nitrogen protection, acetic anhydride (10 mL) and nitric acid (3 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 50 min to prepare a mixed acid solution. Methyl 3-hydroxy-2,2-dimethylpropionate (compound 6a) (4.0 g, 30.3 mmol) was added dropwise, and stirring was continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched by adding water (50 mL). The mixture was extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated by medium pressure to obtain methyl 2,2-dimethyl-3-(nitoxy)propionate (compound 6b) (3.8 g, yield 71%, colorless oil). 1 H NMR (500MHz, CDCl3) δ: 4.50 (s, 2H), 3.71 (s, 3H), 1.28 (s, 6H) ppm.

[0213] Step 2: Preparation of 2,2-dimethyl-3-(nitoxy)propionic acid (compound 6c)

[0214] Under nitrogen protection, methyl 2,2-dimethyl-3-(nitoxy)propionate (compound 6b) (3.8 g, 21.5 mmol), methanol (50 mL), water (10 mL), and sodium hydroxide (1.1 g, 27.5 mmol) were added sequentially to the reaction flask at 0 °C. The mixture was stirred at room temperature for 6 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 2,2-dimethyl-3-(nitoxy)propionate (compound 6c) (2.2 g, yield 63%, white solid). 1 H NMR (500MHz, CDCl3) δ: 4.54 (s, 2H), 1.35 (s, 6H) ppm.

[0215] Step 3: Preparation of methyl chloro2,2-dimethyl-3-(nitoxy)propionate (compound 6d)

[0216] Under nitrogen protection, 2,2-dimethyl-3-(nitoxy)propionic acid (compound 6c) (1.0 g, 6.1 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (2.6 g, 30.9 mmol), and tetrabutylammonium bisulfate (0.21 g, 0.6 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.3 g, 8.7 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 2,2-dimethyl-3-(nitoxy)propionic acid (compound 6d) (1.1 g, 85% yield, colorless oil). 1 H NMR (400MHz, CDCl3) δ: 5.77 (s, 2H), 4.56 (s, 2H), 1.36 (s, 6H) ppm.

[0217] Step 4: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-3-(nitrooxy)propionate (compound 6e)

[0218] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)tert-butyl carbamate (compound M1) (0.5 g, 1.23 mmol), methyltetrahydrofuran (20 mL), methyl 2,2-dimethyl-3-(nitoxy)propionate (compound 6d) (0.34 g, 1.6 mmol), and cesium carbonate (0.45 g, 1.4 mmol) were added sequentially to the reaction flask. The mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was then filtered, dried over anhydrous sodium sulfate, and filtered again. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-3-(nitrooxy)propionate (compound 6e) (0.61 g, yield 86%, white solid). LC-MS m / z (ESI) = 583.5 [M+H] + .

[0219] Step 5: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-3-(nitrooxy)propionate hydrochloride (compound 6)

[0220] A solution of dioxane hydrochloride (2.0 mL, 8.2 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-3-(nitrooxy)propionate (compound 6e) (0.48 g, 0.82 mmol) in ethyl acetate (15 mL). The resulting mixture was stirred at room temperature for 8 hours. The mixture was filtered and dried to give (S)-(5-(((3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-3-(nitrooxy)propionate hydrochloride (compound 6) (390 mg, 91% yield, white solid). 1H NMR(500MHz,DMSO-d6)δ:8.63(dd,1H),8.47(d,3H),8.16(d,1H),7.69(t,1H),7.54(s,1H),5.93(s,2H),4.58(s,2H ),4.08-4.01(m,1H),3.80-3.66(m,2H),3.57-3.46(m,2H),2.58(s,3H),2.45(td,1H),2.18(s,1H),1.20(s,6H)ppm. LC-MS m / z(ESI)=483.5[M+H] + .

[0221] [Amended according to Rule 26, 31.07.2025] Example 7: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)3-(nitrooxy)cyclobutane-1-carboxylic acid methyl ester (compound 7)

[0222] Step 1: Preparation of 3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 7b)

[0223] Under nitrogen protection, acetic anhydride (30 mL) and nitric acid (15 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of acetic anhydride (30 mL) containing 1.5 g (12.9 mmol) of 3-hydroxycyclobutane-1-carboxylic acid (compound 7a) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (50 mL). The mixture was extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 7b) (1.5 g, yield 72%, colorless oil). LC-MS m / z (ESI) = 160.1 [MH] - .

[0224] Step 2: Preparation of chloromethyl 3-(nitrooxy)cyclobutane-1-carboxylate (compound 7c)

[0225] Under nitrogen protection, 3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 7b) (1.5 g, 9.3 mmol), dichloromethane (50 mL), water (50 mL), sodium bicarbonate (3.9 g, 46.5 mmol), and tetrabutylammonium bisulfate (0.32 g, 0.9 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (2.0 g, 13.4 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain chloromethyl 3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 7c) (1.4 g, 72% yield, colorless oil). 1 H NMR (500MHz, CDCl3) δ: 5.73 (s, 2H), 5.20 (p, 1H), 2.93 (dd, 1H), 2.80 (m, 2H), 2.60-2.48 (m, 2H) ppm.

[0226] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(nitrooxy)cyclobutane-1-carboxylate (compound 7d)

[0227] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)tert-butyl carbamate (compound M1) (0.2 g, 0.49 mmol), methyltetrahydrofuran (10 mL), chloromethyl 3-(nitrooxy)cyclobutane-1-carboxylate (compound 7c) (0.15 g, 0.72 mmol), and cesium carbonate (0.19 g, 0.58 mmol) were added sequentially to the reaction flask. The mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was then filtered, dried over anhydrous sodium sulfate, and filtered again. The filtrate was concentrated under reduced pressure, and the residue was separated under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(nitrooxy)cyclobutane-1-carboxylate (compound 7d) (0.23 g, yield 81%, white solid). 1H NMR(500MHz, CDCl3)δ:8.80(d,1H),8.09(d,1H),7.56(t,1H),7.22(s,1H),5.96(s,2H),5.17(p,1H),4.82(s,1H),4.38(s,1H),3.70(dt ,2H),3.55(s,1H),3.44(d,1H),2.96-2.87(m,1H),2.76(m,2H),2.72(s,3H),2.52(m,2H),2.35(td,1H),2.05(dd,1H),1.48(s,9H)ppm. LC-MS m / z(ESI)=581.4[M+H] + .

[0228] Step 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)3-(nitrooxy)cyclobutane-1-carboxylic acid methyl ester (compound 7)

[0229] At room temperature, a solution of dioxane hydrochloride (0.9 mL, 3.4 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(nitrooxy)cyclobutane-1-carboxylate (compound 7d) (0.2 g, 0.34 mmol) in 10 mL of ethyl acetate. The resulting mixture was stirred at room temperature for 8 hours. The mixture was concentrated under reduced pressure, the residue was diluted with saturated sodium bicarbonate solution (50 mL), extracted with ethyl acetate (100 mL × 2), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was prepared and separated under medium pressure to obtain compound (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)3-(nitrooxy)cyclobutane-1-carboxylic acid methyl ester (compound 7) (110 mg, yield 67%, white solid). 1 H NMR(400MHz, CDCl3)δ:8.75(d,1H),8.35(d,1H),7.52(t,1H),7.18(s,1H),5.94(s,2H),5.14(p,1H),3.91-3.80(m,1H ),3.66(m,3H),3.24(dd,1H),2.94-2.84(m,1H),2.81-2.64(m,6H),2.50(m,2H),2.29-2.19(m,2H),1.87(dt,1H)ppm. LC-MS m / z(ESI)=481.4[M+H] + .

[0230] [Amended according to Rule 26, 31.07.2025] Example 8: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid hydrochloride (compound 8)

[0231] Step 1: Preparation of 5,5-bis(bromomethyl)-2-phenyl-1,3-dioxane (compound 8b)

[0232] Under nitrogen protection, 2,2-dibromomethylpropane-1,3-diol (compound 8a) (25.0 g, 95.5 mmol), toluene (300 mL), benzaldehyde (11.1 g, 104.6 mmol), and p-toluenesulfonic acid (1.6 g, 9.5 mmol) were added sequentially to the reaction flask, and the mixture was heated to reflux for 6 hours. The reaction was monitored by TLC until complete. The mixture was cooled to room temperature, washed with saturated sodium bicarbonate aqueous solution (200 mL) and saturated brine (200 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 5,5-bis(bromomethyl)-2-phenyl-1,3-dioxane (compound 8b) (25 g, 75% yield, white solid). 1 H NMR (500MHz, CDCl3) δ: 7.48 (dd, 2H), 7.42-7.33 (m, 3H), 5.41 (s, 1H), 4.26 (d, 2H), 3.99 (s, 2H), 3.86 (d, 2H), 3.32 (s, 2H) ppm. LC-MS m / z(ESI)=351.1[M+H] + .

[0233] Step 2: Preparation of diethyl 7-phenyl-6,8-dioxane[3.5]nonane-2,2-dicarboxylate (compound 8c)

[0234] Under nitrogen protection, sodium hydride (5.7 g, 142.3 mmol, 60%) was added in portions to diethyl malonate (22.9 g, 142.3 mmol) in N,N-dimethylformamide (150 mL) at 0 °C, and the mixture was stirred for 20 min. Then, a solution of 5,5-bis(bromomethyl)-2-phenyl-1,3-dioxane (compound 8b) (25.0 g, 71.4 mmol) in N,N-dimethylformamide (100 mL) was added dropwise. After the addition was complete, the mixture was heated to 140 °C and stirred for 3 hours. The reaction was monitored by TLC until it was complete. The mixture was then cooled to 0 °C, and a saturated ammonium chloride solution (200 mL) was added dropwise to quench the reaction. Extracted with ethyl acetate (200 mL × 2), separated, the organic phase was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated under medium pressure to obtain compound 7-phenyl-6,8-dioxane[3.5]nonane-2,2-dicarboxylic acid diethyl ester (compound 8c) (11 g, yield 44%, white solid). 1 H NMR(500MHz, CDCl3)δ:7.51-7.45(m,2H),7.42-7.32(m,3H),5.43(s,1H),4.29- 4.21(m,4H),4.17(d,2H),3.78(d,2H),2.82(s,2H),2.25(s,2H),1.29(dd,6H). LC-MS m / z(ESI)=349.3[M+H] + .

[0235] Step 3: Preparation of 7-phenyl-6,8-dioxane[3.5]nonane-2,2-dicarboxylic acid (compound 8d)

[0236] Under nitrogen protection, diethyl 7-phenyl-6,8-dioxaspiro[3.5]nonane-2,2-dicarboxylic acid (compound 8c) (11.7 g, 33.6 mmol), ethanol (215 mL), water (125 mL), and sodium hydroxide (4.0 g, 100.8 mmol) were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 4 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and the pH was adjusted to 2 with 1 N hydrochloric acid. The mixture was then filtered. The filter cake was dried to give compound 7-phenyl-6,8-dioxaspiro[3.5]nonane-2,2-dicarboxylic acid (compound 8d) (8.1 g, 82% yield, white solid). 1 H NMR(400MHz, DMSO-d6)δ:12.91(s,2H),7.46-7.29(m,5H),5.46(s,1H),3.98(d,2H),3.77(d,2H),2.54(s,2H),2.12(s,2H)ppm. LC-MS m / z(ESI)=291.2[MH] - .

[0237] Step 4: Preparation of 7-phenyl-6,8-dioxane[3.5]nonane-2-carboxylic acid (compound 8e)

[0238] At room temperature, 7-phenyl-6,8-dioxaspiro[3.5]nonane-2,2-dicarboxylic acid (compound 8d) (8.1 g, 27.7 mmol) and pyridine (150 mL) solution were added sequentially to a reaction flask, and the mixture was heated to reflux for 21 h. The mixture was then cooled to room temperature, concentrated under reduced pressure, and the pH was adjusted to 5 with 1 N hydrochloric acid. Extraction was performed with ethyl acetate (200 mL × 2), and the organic phase was separated. The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain compound 7-phenyl-6,8-dioxaspiro[3.5]nonane-2-carboxylic acid (compound 8e) (6.5 g, 92% yield, white solid). 1 H NMR(400MHz,DMSO-d6)δ:12.32(s,1H),7.39-7.32(m,5H),5.43(s,1H),4.11(dd,1H),4.01-3.90(m ,1H),3.76-3.65(m,2H),3.13-3.02(m,1H),2.31(t,1H),2.16-2.05(m,1H),1.83-1.72(m,2H)ppm. LC-MS m / z(ESI)=247.2[MH] - .

[0239] Step 5: Preparation of 3,3-bis(hydroxymethyl)cyclobutane-1-carboxylic acid (compound 8f)

[0240] At room temperature, 7-phenyl-6,8-dioxane[3.5]nonane-2-carboxylic acid (compound 8e) (6.5 g, 26.2 mmol), methanol (150 mL), and palladium on carbon (1.8 g, 10%) were added sequentially to a reaction flask, and the reaction was hydrogenated at room temperature for 15 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give compound 3,3-bis(hydroxymethyl)cyclobutane-1-carboxylic acid (compound 8f) (4.3 g, 100% yield, colorless oil).

[0241] Step 6: Preparation of 3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 8g)

[0242] Under nitrogen protection, acetic anhydride (15 mL) and nitric acid (7.5 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of 3,3-bis(hydroxymethyl)cyclobutane-1-carboxylic acid (compound 8f) (1.0 g, 6.24 mmol) in acetic anhydride (50 mL) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The reaction was quenched with water (100 mL). The mixture was extracted with ethyl acetate (100 mL × 3), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 8 g) (1.0 g, yield 64%, white solid). LC-MS m / z (ESI) = 249.1 [MH] - .

[0243] Step 7: Preparation of chloromethyl 3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylate (compound 8h)

[0244] Under nitrogen protection, 3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 8 g) (1.0 g, 4.0 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (1.7 g, 20.0 mmol), and tetrabutylammonium bisulfate (0.14 g, 0.4 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.0 g, 6.7 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain chloromethyl 3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 8 h) (0.9 g, yield 76%, colorless oil). 1 H NMR (500MHz, CDCl3) δ: 5.75 (s, 2H), 4.55 (d, 4H), 3.30 (m, 1H), 2.43-2.31 (m, 4H) ppm.

[0245] Step 8: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid ester (compound 8i)

[0246] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)tert-butyl carbamate (compound M1) (0.3 g, 0.73 mmol), methyltetrahydrofuran (10 mL), chloromethyl 3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylate (compound 8h) (0.33 g, 1.1 mmol), and cesium carbonate (0.29 g, 0.88 mmol) were added sequentially to the reaction flask. The mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was then filtered, dried over anhydrous sodium sulfate, and filtered again. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid ester (compound 8i) (0.42 g, yield 86%, white solid). LC-MS m / z (ESI) = 670.4 [M+H] + .

[0247] Step 9: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid hydrochloride (compound 8)

[0248] A solution of dioxane hydrochloride (1.5 mL, 6.0 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid ester (compound 8i) (0.40 g, 0.60 mmol) in ethyl acetate (15 mL). The resulting mixture was stirred at room temperature for 8 hours. The mixture was filtered and dried to give compound (S)-(5-((3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3,3-bis((nitoxy)methyl)cyclobutane-1-carboxylic acid hydrochloride (compound 8) (260 mg, 72% yield, white solid). 1H NMR(400MHz,DMSO-d6)δ8.62(dd,1H),8.50(s,3H),8.16(dd,1H),7.68(t,1H),7.55(s,1H),5.90(s,2H),4.64(s,2H ),4.51(s,2H),3.78-3.65(m,2H),3.51(m,2H),3.33-3.27(m,1H),2.57(s,3H),2.42(dt,1H),2.32-2.10(m,6H)ppm. LC-MS m / z(ESI)=570.4[M+H] + .

[0249] [Amended according to Rule 26, 31.07.2025] Example 9: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester hydrochloride (compound 9)

[0250] Step 1: Preparation of methyl 3-hydroxymethylbicyclo[1.1.1]pentane-1-carboxylic acid (compound 9b)

[0251] 3-(methoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9a) (4.5 g, 26.4 mmol) was added to a 250 mL three-necked flask, followed by the addition of tetrahydrofuran (50 mL). Under nitrogen protection, a boranetetrahydrofuran solution (31.7 mL, 31.7 mmol) was added and the mixture was stirred for 3 hours in an ice bath. The reaction was monitored by TLC until complete, and then quenched with water. The mixture was concentrated under reduced pressure, extracted with ethyl acetate (100 mL), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was then subjected to preparative separation under medium pressure to obtain methyl 3-hydroxymethylbicyclo[1.1.1]pentane-1-carboxylic acid (compound 9b) (3.5 g, 85% yield, colorless oil). 1 H NMR (500MHz, CDCl3) δ: 3.70 (s, 3H), 3.65 (s, 2H), 2.02 (s, 6H) ppm.

[0252] Step 2: Preparation of 3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9c)

[0253] Methyl 3-hydroxymethylbicyclo[1.1.1]pentane-1-carboxylic acid (compound 9b) (3.4 g, 21.8 mmol) was added to a 250 mL single-necked flask, followed by methanol (60 mL), tetrahydrofuran (60 mL), and sodium hydroxide aqueous solution (21.8 mL, 43.5 mmol, 2N). The mixture was stirred at room temperature for 5 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, diluted with water (50 mL), and the pH was adjusted to 6–7 with hydrochloric acid. The aqueous layer was extracted with ethyl acetate (100 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then subjected to preparative separation under medium pressure to obtain compound 3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9c) (1.7 g, 55% yield, colorless oil). 1 H NMR (400MHz, DMSO-d6) δ: 3.37-3.34 (m, 2H), 1.82 (s, 6H) ppm.

[0254] Step 3: Preparation of 3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9d)

[0255] Under ice bath conditions, nitric acid (17 mL) was slowly added dropwise to concentrated sulfuric acid (17 mL), and the mixture was stirred for 10 minutes. 3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9c) (1.7 g, 11.9 mmol) was dissolved in dichloromethane (50 mL) and then slowly added to the above mixed solution. The mixture was stirred under ice bath conditions for 2 hours. The reaction was monitored by TLC until complete. The reaction solution was slowly quenched in ice water, extracted with ethyl acetate (50 mL), washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was then subjected to preparative separation under medium pressure to obtain compound 3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9d) (1.7 g, yield 76%, colorless oil). 1 H NMR (500MHz, CDCl3) δ4.50 (s, 2H), 2.13 (s, 6H) ppm.

[0256] Step 4: 3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 9e)

[0257] 3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9d) (700 mg, 3.7 mmol) was added to a 100 mL two-necked flask, along with dichloromethane (10 mL), water (10 mL), sodium bicarbonate (1.6 g, 18.7 mmol), and tetrabutylammonium bisulfate (254 mg, 0.75 mmol). Under nitrogen protection, chloromethyl chlorosulfonate (0.46 mL, 4.5 mmol) was added in an ice bath, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. After standing, the mixture was separated, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. After separation under medium pressure, compound 3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 9e) (690 mg, yield 78%, white solid) was obtained. 1 H NMR (400MHz, CDCl3) δ: 5.73 (s, 2H), 4.52 (s, 2H), 2.20 (s, 6H) ppm.

[0258] Step 5: (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester (compound 9f)

[0259] (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (579 mg, 1.42 mmol) was added to a 100 mL single-necked flask, followed by 2-methyltetrahydrofuran (20 mL), 3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 9e) (690 mg, 2.93 mmol), and cesium carbonate (694 mg, 2.13 mmol). The mixture was stirred at 55 °C for 3 hours under nitrogen protection. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, dissolved in dichloromethane (50 mL), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was then subjected to preparative separation under medium pressure to yield compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester (compound 9f) (660 mg, yield 76%, white solid). LC-MS m / z (ESI) = 607.2 [M+H] + .

[0260] Step 6: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester hydrochloride (compound 9)

[0261] Methyl (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 9f) (610 mg, 1.0 mmol) was added to a 50 mL single-necked flask, along with acetonitrile (10 mL) and p-toluenesulfonic acid (689 mg, 4.0 mmol). The mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC until complete, and the solution was concentrated under reduced pressure to obtain the crude product. The crude product was separated under high pressure to obtain a pure product. Ethyl acetate (10 mL) and dioxane hydrochloride solution (1.0 mL, 4 N) were added, and the mixture was stirred at room temperature for 10 minutes. After filtration, compound (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)3-((nitrooxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester hydrochloride (compound 9) (200 mg, yield 39%, white solid). 1 H NMR(400MHz,DMSO-d6)δ:8.65(d,1H),8.53(s,3H),8.19(d,1H),7.71(t,1H),7.57(s,1H),5.91(s,2H),4.59 (s,2H),3.82-3.66(m,2H),3.59-3.46(m,3H),2.58(d,3H),2.48-2.40(m,1H),2.20(d,1H),2.04(s,6H)ppm. LC-MS m / z(ESI)=507.1[M+H] + .

[0262] [Amended according to Rule 26, 31.07.2025] Example 10: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-3-(nitrooxy)-2-((nitrooxy)methyl)propionate hydrochloride (compound 10)

[0263] Step 1: Preparation of 2-methyl-3-(nitoxy)-2-((nitoxy)methyl)propionic acid (compound 10b)

[0264] Under nitrogen protection, acetic anhydride (30 mL) and nitric acid (6 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of ethyl acetate (150 mL) containing 1.5 g (11.2 mmol) of 3-hydroxy-2-hydroxymethyl-2-methylpropionic acid (compound 10a) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (50 mL). The mixture was extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain compound 2-methyl-3-(nitoxy)-2-((nitoxy)methyl)propionic acid (compound 10b) (2.8 g, yield 67%, white solid). LC-MS m / z (ESI) = 223.1 [MH] - .

[0265] Step 2: Preparation of chloromethyl 2-methyl-3-(nitoxy)-2-((nitoxy)methyl)propionate (compound 10c)

[0266] Under nitrogen protection, 2-methyl-3-(nitoxy)-2-((nitoxy)methyl)propionic acid (compound 10b) (2.0 g, 8.9 mmol), dichloromethane (50 mL), water (50 mL), sodium bicarbonate (3.8 g, 45.2 mmol), and tetrabutylammonium bisulfate (0.3 g, 0.9 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethylsulfonyl chloride (1.9 g, 12.8 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to obtain chloromethyl 2-methyl-3-(nitoxy)-2-((nitoxy)methyl)propionic acid (compound 10c) (2.1 g, 87% yield, colorless oil). 1 H NMR (400MHz, CDCl3) δ: 5.80 (s, 2H), 4.74 (d, 2H), 4.66 (d, 2H), 1.45 (s, 3H) ppm.

[0267] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-3-(nitro)-2-((nitro)methyl)propionate (compound 10d)

[0268] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)tert-butyl carbamate (compound M1) (0.3 g, 0.73 mmol), methyltetrahydrofuran (10 mL), methyl 2-methyl-3-(nitoxy)-2-((nitoxy)methyl)propionate (compound 10c) (0.3 g, 1.1 mmol), and cesium carbonate (0.29 g, 0.88 mmol) were added sequentially to the reaction flask. The mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was then filtered, dried over anhydrous sodium sulfate, and filtered again. The filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-3-(nitrooxy)-2-((nitrooxy)methyl)propionate (compound 10d) (0.38 g, yield 81%, white solid). LC-MS m / z (ESI) = 644.5 [M+H] + .

[0269] Step 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-3-(nitrooxy)-2-((nitrooxy)methyl)propionate hydrochloride (compound 10)

[0270] A solution of dioxane hydrochloride (1.5 mL, 6.0 mmol, 4N) was added dropwise to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-3-(nitrooxy)-2-((nitrooxy)methyl)propionate (compound 10d) (0.38 g, 0.60 mmol) in ethyl acetate (15 mL). The resulting mixture was stirred at room temperature for 8 hours. The mixture was filtered and dried to give (S)-(5-((3-aminopyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-methyl-3-(nitrooxy)-2-((nitrooxy)methyl)propionate hydrochloride (compound 10) (310 mg, 91% yield, white solid). 1H NMR(400MHz,DMSO-d6)δ:8.63(dd,1H),8.51(s,3H),8.18(dd,1H),7.71(t,1H),7.55(s,1H),5.99(s,2H),4.72(q ,4H),3.82-3.65(m,2H),3.59(s,1H),3.53(m,2H),2.59(s,3H),2.46(dd,1H),2.24-2.15(m,1H),1.29(s,3H)ppm. LC-MS m / z(ESI)=544.5[M+H] + .

[0271] Example 11: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]1-carboxylic acid pentane hydrochloride (compound 11)

[0272] Step 1: Preparation of 3-((2-hydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 11a)

[0273] At 0°C, under nitrogen protection, a solution of borane tetrahydrofuran (15.3 mL, 15.3 mmol, 1N) was added dropwise to a solution of methyl 3-((allyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 34a) (2.00 g, 10.19 mmol) in tetrahydrofuran (20 mL). The resulting mixture was heated to room temperature and stirred for 1 hour. The mixture was cooled to 0°C, and sodium hydroxide aqueous solution (1.43 g, 35.67 mmol, 2N) and hydrogen peroxide (3.47 g, 30.57 mmol, 30%) were added dropwise. After the addition was complete, the reaction was maintained at this temperature for 30 min, then returned to room temperature and stirred for 1 h. The mixture was then cooled to 0°C, and the reaction was quenched by adding saturated sodium bisulfite solution (100 mL). The pH was adjusted to 2 with hydrochloric acid aqueous solution, and the mixture was extracted with ethyl acetate (200 mL × 2). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 11a (1.8 g, 88% yield, colorless oil). LC-MS m / z (ESI) = 199.2 [MH] - .

[0274] Step 2: Preparation of 3-((2-(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 11b)

[0275] Under nitrogen protection, dichloromethane (50 mL), concentrated nitric acid (17 mL), and concentrated sulfuric acid (17 mL) were added sequentially to the reaction flask at 0 °C. The mixture was stirred for 30 min to prepare a mixed acid solution. A solution of 3-((2-hydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 11a) (1.80 g, 8.99 mmol) in dichloromethane (20 mL) was added dropwise, and stirring was continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The reaction was quenched with water (50 mL), and the mixture was extracted with dichloromethane (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparative chromatography to obtain compound 11b (1.6 g, yield 73%, colorless oil). LC-MS m / z (ESI) = 244.2 [MH] - .

[0276] Step 3: Preparation of 3-((2-(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 11c)

[0277] 3-((2-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 11b) (1.60 g, 6.52 mmol), dichloromethane (40 mL), water (40 mL), sodium bicarbonate (2.74 g, 32.62 mmol) and tetrabutylammonium bisulfate (0.44 g, 1.31 mmol), with chloromethylsulfonyl chloride (1.61 g, 9.79 mmol) added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, the organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by medium pressure to give compound 11c (1.6 g, 84% yield, colorless oil).

[0278] Step 4: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester (compound 11d)

[0279] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), 3-((2-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 11c) (0.54 g, 1.84 mmol), and cesium carbonate (0.47 g, 1.47 mmol) were added sequentially to the reaction flask. The mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 11d (0.10 g, 12% yield, white solid). LC-MS m / z (ESI) = 665.6 [M+H] + .

[0280] Step 5: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]1-carboxylic acid pentane hydrochloride (compound 11)

[0281] At room temperature, p-toluenesulfonic acid hydrate (114.50 mg, 0.60 mmol) was added to a 5 mL solution of compound 11d (0.10 g, 0.15 mmol) in acetonitrile, and stirred for 0.5 h at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 with sodium bicarbonate solution. The aqueous phase was extracted with 50 mL of ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate. The mixture was filtered, concentrated under reduced pressure, and the residue was separated under medium pressure. The product was added to 0.5 mL of ethyl acetate hydrochloride solution (4 N), and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]1-carboxylic acid pentane hydrochloride (compound 11) (52 mg, 57% yield, white solid). 1H NMR(400MHz,DMSO-d6)δ8.67-8.57(m,4H),8.36(d,1H),7.54(t,1H),7.22(s,1H),5.94(s,2H),5.32(s,2H),3.91-3.86(m,1H),3.73- 3.64(m,2H),3.55(d,2H),3.51-3.42(m,2H),3.26(m,1H),2.73(s,3H),2.31-2.21(m,1H),2.04(s,6H),1.89(m,1H),1.35(d,3H)ppm. LC-MS m / z(ESI)=565.6[M+H] + .

[0282] Example 12: Preparation of methyl 3-(5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2,3-bis(nitoxy)propoxy)-2-propionate hydrochloride (compound 12)

[0283] Step 1: Preparation of (R)-3-(allyloxy)-2-methylpropionate (compound 12b)

[0284] Under nitrogen protection, methyl (R)-3-hydroxy-2-methylpropionate (compound 12a) (20.0 g, 0.169 mol), petroleum ether (800 mL), anhydrous magnesium sulfate (24.40 g, 0.20 mol), and silver oxide (51.00 g, 0.22 mol) were added sequentially to the reaction flask at 0 °C. Allyl bromide (30.70 g, 0.25 mol) was added dropwise, and the mixture was stirred at room temperature for 8 hours. The reaction was monitored by TLC until complete. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by medium-pressure preparative separation to obtain compound (R)-3-(allyloxy)-2-methylpropionate (compound 12b) (20 g, 75% yield, colorless oil). 1 H NMR(500MHz, CDCl3)δ5.87(m,1H),5.27-5.21(m,1H),5.16(d,1H),3.97(d, 2H),3.69(s,3H),3.62(t,1H),3.45(dd,1H),2.75(dd,1H),1.17(d,3H)ppm. LC-MS m / z(ESI)=159.2[M+H] + .

[0285] Step 2: Preparation of methyl (2R)-3-(2,3-dihydroxypropoxy)-2-methylpropionate (compound 12c)

[0286] Under nitrogen protection, tert-butanol (300 mL), purified water (300 mL), and AD-mix-α (50 g) were added sequentially to the reaction flask. The mixture was stirred at room temperature for ten minutes, then cooled to 0 °C. Methyl (R)-3-(allyloxy)-2-methylpropionate (compound 12b) (5.0 g, 31.6 mmol) was added dropwise, and the reaction was stirred for 4 hours. The reaction was monitored by TLC until complete. The reaction was quenched by adding sodium sulfite aqueous solution (100 mL). The aqueous phase was extracted with ethyl acetate (200 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then subjected to preparative separation under medium pressure to obtain compound (2R)-3-(2,3-dihydroxypropoxy)-2-methylpropionate (compound 12c) (2.7 g, yield 44%, colorless oil). LC-MS m / z (ESI) = 193.2 [M+H] + .

[0287] Step 3: Preparation of methyl (2R)-3-(2,3-bis(nitoxy)propoxy)-2-methylpropionate (compound 12d)

[0288] Under nitrogen protection, acetic anhydride (80 mL) and concentrated nitric acid (20 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 minutes to prepare a mixed acid solution. A solution of ethyl acetate (50 mL) containing methyl (2R)-3-(2,3-dihydroxypropoxy)-2-methylpropionate (compound 12c) (2.7 g, 14 mmol) was added dropwise, and stirring was continued for 0.5 hours after the addition was complete. The reaction was monitored by TLC until it was complete. The mixture was cooled to 0 °C, and the reaction was quenched with water (200 mL). The mixture was separated, extracted with ethyl acetate (200 mL × 2), and the combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain methyl (2R)-3-(2,3-bis(nitoxy)propoxy)-2-methylpropionate (compound 12d) (3.5 g, yield 88%, colorless oil). LC-MS m / z(ESI) = 283.2 [M+H] + .

[0289] Step 4: Preparation of 3-(2,3-bis(nitoxy)propoxy)-2-methylpropionic acid (compound 12e)

[0290] Under nitrogen protection, methyl (2R)-3-(2,3-bis(nitoxy)propoxy)-2-methylpropionic acid (compound 12d) (3.5 g, 12.4 mmol), tetrahydrofuran (50 mL), water (5 mL), and lithium hydroxide (0.59 g, 24.80 mmol) were added sequentially to the reaction flask at 0 °C. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (100 mL × 3), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 3-(2,3-bis(nitoxy)propoxy)-2-methylpropionic acid (compound 12e) (2.0 g, 60% yield, colorless oil). LC-MS m / z (ESI) = 267.1 [MH] - .

[0291] Step 5: Preparation of chloromethyl 3-(2,3-bis(nitoxy)propoxy)-2-methylpropionate (compound 12f)

[0292] Under nitrogen protection, 3-(2,3-bis(nitoxy)propoxy)-2-methylpropionic acid (compound 12e) (2.0 g, 7.5 mmol), dichloromethane (40 mL), water (40 mL), sodium bicarbonate (3.1 g, 37.3 mmol), and tetrabutylammonium bisulfate (0.51 g, 1.50 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.6 g, 9.7 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 3-(2,3-bis(nitoxy)propoxy)-2-methylpropionic acid (compound 12f) (1.7 g, yield 72%, colorless oil). 1 H NMR (400MHz, CDCl3) δ5.72(s,2H),5.38(dd,1H),4.79(dt,1H),4.61(dd,1H),3.79-3.56(m,4H),2.81(m,1H),1.20(dd,3H)ppm.

[0293] Step 6: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2,3-bis(nitrooxy)propoxy)-2-methylpropionate (compound 12g)

[0294] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.5 g, 1.2 mmol), methyltetrahydrofuran (10 mL), methyl 3-(2,3-bis(nitoxy)propoxy)-2-methylpropionate (compound 12f) (0.58 g, 1.80 mmol) and cesium carbonate (0.48 g, 1.50 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2,3-bis(nitrooxy)propoxy)-2-methylpropionate (12 g of compound) (0.61 g, yield 73%, white solid). LC-MS m / z (ESI) = 688.4 [M+H] + .

[0295] Step 7: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2,3-bis(nitoxy)propoxy)-2-propionate methyl hydrochloride (compound 12)

[0296] p-Toluenesulfonic acid hydrate (0.67 g, 3.55 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2,3-bis(nitrooxy)propoxy)-2-methylpropionate (compound 12 g) (0.61 g, 0.89 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give methyl 5-(5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 3-(2,3-bis(nitoxy)propoxy)-2-propanoate hydrochloride (compound 12) (510 mg, yield 92%, white solid). 1H NMR(400MHz,DMSO-d6)δ8.67-8.40(m,4H),8.16(d,1H),7.67(t,1H),7.52(s,1H),5.89(s,2H),5.55-5.47(m,1H),4.89-4.79(m,1H), 4.66(m,1H),4.02(m,1H),3.80-3.66(m,4H),3.61-3.49(m,4H),2.76(m,1H),2.57(s,3H),2.43(m,1H),2.18(m,1H),1.04(d,3H)ppm. LC-MS m / z(ESI)=588.2[M+H] + .

[0297] [Amended according to Rule 26, 31.07.2025] Example 13: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate hydrochloride (compound 13)

[0298] Step 1: Preparation of methyl 2-methyl-4-((2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilundecane-6-yl)oxy)benzoate (compound 13b)

[0299] Under nitrogen protection, diisopropyl azodicarbonate (12.2 g, 60.2 mmol) was added dropwise to methyl 4-hydroxy-2-methylbenzoate (compound 13a) (5.0 g, 30.1 mmol), 2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilundecane-6-ol (19.3 g, 60.2 mmol), triphenylphosphine (11.8 g, 45.1 mmol), and tetrahydrofuran (100 mL) at 0 °C, and stirred at room temperature for 3 hours. The mixture was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 2-methyl-4-((2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilundecane-6-yl)oxy)benzoate (compound 13b) (14 g, 99% yield, colorless oil). LC-MS m / z (ESI) = 469.5 [M+H] + .

[0300] Step 2: Preparation of methyl 4-((1,3-dihydroxypropyl-2-yl)oxy)-2-methylbenzoate (13c)

[0301] Under nitrogen protection, tetrabutylammonium fluoride (12.8 mL, 12.8 mmol, 1N) was added dropwise to methyl 2-methyl-4-((2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilundec-6-yl)oxy)benzoate (compound 13b) (3.0 g, 6.4 mmol) in tetrahydrofuran (30 mL), and stirred at room temperature for 0.5 h. The mixture was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give methyl 4-((1,3-dihydroxypropyl-2-yl)oxy)-2-methylbenzoate (compound 13c) (0.7 g, 46% yield, colorless oil). LC-MS m / z (ESI) = 241.2 [M+H] + .

[0302] Step 3: Preparation of 4-((1,3-dihydroxypropyl-2-yl)oxy)-2-methylbenzoic acid (compound 13d)

[0303] Under nitrogen protection, methyl 4-((1,3-dihydroxypropyl-2-yl)oxy)-2-methylbenzoate (13c) (1.3 g, 5.4 mmol), methanol (20 mL), water (10 mL), and potassium hydroxide (1.2 g, 21.6 mmol) were added sequentially to the reaction flask. The mixture was heated to 60 °C and stirred for 8 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (150 mL × 3), and the combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 4-((1,3-dihydroxypropyl-2-yl)oxy)-2-methylbenzoic acid (compound 13d) (0.75 g, yield 61%, white solid). LC-MS m / z (ESI) = 225.2 [MH] - .

[0304] Step 4: Preparation of 4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoic acid (compound 13e) Under nitrogen protection, dichloromethane (50 mL), concentrated nitric acid (8 mL), and concentrated sulfuric acid (8 mL) were added sequentially to the reaction flask at 0 °C. The mixture was stirred for 30 minutes to prepare a mixed acid solution. A solution of 4-((1,3-dihydroxyprop-2-yl)oxy)-2-methylbenzoic acid (compound 13d) (2.0 g, 8.8 mmol) in dichloromethane (50 mL) was then added dropwise. After the addition was complete, stirring continued for 0.5 hours; the reaction was monitored by TLC until complete; the temperature was lowered to 0℃, and the reaction was quenched with water (100 mL). The mixture was separated, extracted with dichloromethane (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoic acid (compound 13e) (0.51 g, yield 18%, colorless oil). LC-MS m / z (ESI) = 315.2 [MH] - .

[0305] Step 5: Preparation of chloromethyl 4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate (compound 13f) Under nitrogen protection, at 0°C, 4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoic acid (compound 13e) (0.50 g, 1.58 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (0.66 g, 7.90 mmol), and tetrabutylammonium bisulfate (0.05 g) were added sequentially to the reaction flask. Add chloromethylsulfonyl chloride (0.39 g, 2.37 mmol) dropwise, and stir at room temperature for 1 hour after the addition is complete; monitor the reaction for completeness by TLC; separate the liquid and liquid phase, wash the organic phase with saturated brine (30 mL), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate the residue under medium pressure to obtain compound 4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate chloromethyl ester (compound 13f) (0.43 g, yield 74%, colorless oil). 1 H NMR (500MHz, CDCl3) δ8.04(d,1H),6.88-6.80(m,2H),5.95(s,2H),4.91(m,1H),4.77(d,4H),2.66(s,3H)ppm.

[0306] Step 6: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate (compound 13g)

[0307] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.30 g, 0.74 mmol), methyltetrahydrofuran (10 mL), methyl 4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate (compound 13f) (0.40 g, 1.1 mmol) and cesium carbonate (0.29 g, 0.88 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate (13 g of compound) (0.43 g, yield 80%, white solid). LC-MS m / z (ESI) = 736.5 [M+H] + .

[0308] Step 7: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate hydrochloride (compound 13)

[0309] p-Toluenesulfonic acid hydrate (0.44 g, 2.34 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate (compound 13 g) (0.43 g, 0.58 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (100 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate hydrochloride (compound 13) (302 mg, yield 77%, white solid). 1 H NMR(400MHz,DMSO-d6)δ8.69(d,1H),8.42(s,3H),8.17(d,1H),7.85(d,1H),7.74-7.69(m,2H),7.04(d,1H),6.97(m,1H),6.11(s,2H ),4.95(dd,2H),4.83(dd,2H),4.06(m,1H),3.82-3.67(m,2H),3.53(m,3H),2.63(s,3H),2.53(s,3H),2.46(m,1H),2.19(m,1H)ppm. LC-MS m / z(ESI)=636.3[M+H] + .

[0310] [Amended according to Rule 26, 31.07.2025] Example 14: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)-2-methylbenzoate hydrochloride (compound 14)

[0311] Step 1: Preparation of 5-(bromomethyl)-2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disildecane (compound 14b)

[0312] Under nitrogen protection, tert-butyldimethylchlorosilane (10.5 g, 69.4 mmol) was added to 3-bromopropane-1,2-diol (compound 14a) (4.3 g, 27.7 mmol), imidazole (7.56 g, 111 mmol), and dichloromethane (100 mL) at 0 °C, and stirred at room temperature for 2 hours. The mixture was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 5-(bromomethyl)-2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disildecane (compound 14b) (10 g, 94% yield, colorless oil).

[0313] Step 2: Preparation of methyl 4-(2,3-bis((tert-butyldimethylsilyl)oxy)propoxy)-2-methylbenzoate (compound 14c)

[0314] Under nitrogen protection, methyl 4-hydroxy-2-methylbenzoate (3.5 g, 21.1 mmol), 5-(bromomethyl)-2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disilylcane (compound 14b) (9.7 g, 25.3 mmol), potassium carbonate (3.5 g, 25.3 mmol), and N,N-dimethylformamide (50 mL) were added sequentially to a reaction flask. The mixture was heated to 100 °C and stirred for 4 hours. The mixture was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 4-(2,3-bis((tert-butyldimethylsilyl)oxy)propoxy)-2-methylbenzoate (compound 14c) (5.3 g, 54% yield, colorless oil). LC-MS m / z (ESI) = 469.7 [M+H] + .

[0315] Step 3: Preparation of methyl 4-(2,3-dihydroxypropoxy)-2-methylbenzoate (compound 14d)

[0316] Under nitrogen protection, tetrabutylammonium fluoride (13.3 mL, 13.3 mmol, 1N) was added dropwise to methyl 4-(2,3-bis((tert-butyldimethylsilyl)oxy)propoxy)-2-methylbenzoate (compound 14c) (5.2 g, 11.1 mmol) in tetrahydrofuran (50 mL), and stirred at room temperature for 0.5 hours. The mixture was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give methyl 4-(2,3-dihydroxypropoxy)-2-methylbenzoate (compound 14d) (2.33 g, 87% yield, colorless oil). LC-MS m / z (ESI) = 241.2 [M+H] + .

[0317] Step 4: Preparation of 4-(2,3-dihydroxypropoxy)-2-methylbenzoic acid (compound 14e)

[0318] Under nitrogen protection, methyl 4-(2,3-dihydroxypropoxy)-2-methylbenzoate (compound 14d) (0.7 g, 2.9 mmol), methanol (10 mL), water (2 mL), and sodium hydroxide (0.23 g, 5.83 mmol) were added sequentially to the reaction flask. The mixture was heated to 60 °C and stirred for 4 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 4-(2,3-dihydroxypropoxy)-2-methylbenzoic acid (compound 14e) (0.6 g, 91% yield, white solid). 1 H NMR(500MHz,DMSO-d6)δ12.43(s,1H),7.83(d,1H),6.83(d,2H),4.97(s,1H),4.68( s,1H),4.05(dd,1H),3.91(dd,1H),3.86-3.75(m,1H),3.45(d,2H),2.52(s,3H)ppm. LC-MS m / z(ESI)=225.2[MH] - .

[0319] Step 5: Preparation of 4-(2,3-bis(nitoxy)propoxy)-2-methylbenzoic acid (compound 14f)

[0320] Under nitrogen protection, dichloromethane (10 mL), concentrated nitric acid (8 mL), and concentrated sulfuric acid (2 mL) were added sequentially to the reaction flask at 0 °C. The mixture was stirred for 30 minutes to prepare a mixed acid solution. A solution of 4-(2,3-dihydroxypropoxy)-2-methylbenzoic acid (compound 14e) (0.55 g, 2.43 mmol) in dichloromethane (10 mL) was added dropwise. After the addition was complete, the mixture was stirred for 0.5 hours. The reaction was monitored by TLC until it was complete. The mixture was cooled to 0 °C, and the reaction was quenched with water (30 mL). The mixture was separated, extracted with dichloromethane (50 mL × 2), and the combined organic phases were washed with saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain compound 4-(2,3-bis(nitoxy)propoxy)-2-methylbenzoic acid (compound 14f) (0.75 g, 97% yield, colorless oil). LC-MS m / z (ESI) = 315.2 [MH] - .

[0321] Step 6: Preparation of chloromethyl 4-(2,3-bis(nitoxy)propoxy)-2-methylbenzoate (compound 14g)

[0322] Under nitrogen protection, 4-(2,3-bis(nitoxy)propoxy)-2-methylbenzoic acid (compound 14f) (0.70 g, 2.21 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (0.93 g, 11.10 mmol), and tetrabutylammonium bisulfate (0.08 g, 0.22 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (0.55 g, 3.32 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 4-(2,3-bis(nitoxy)propoxy)-2-methylbenzoate (compound 14 g) (0.13 g, yield 16%, colorless oil).

[0323] Step 7: Preparation of methyl 4-(5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)-2-benzoate (compound 14h)

[0324] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.20 g, 0.49 mmol), methyltetrahydrofuran (5 mL), chloromethyl 4-(2,3-bis(nitoxy)propoxy)-2-methylbenzoate (compound 14 g) (0.21 g, 0.58 mmol) and cesium carbonate (0.19 g, 0.59 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)-2-benzoate (compound 14h) (0.3 g, yield 83%, white solid). LC-MS m / z (ESI) = 736.5 [M+H] + .

[0325] Step 8: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)-2-methylbenzoate hydrochloride (compound 14)

[0326] p-Toluenesulfonic acid hydrate (0.21 g, 1.09 mmol) was added to a solution of methyl 5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)-2-benzoate (compound 14h) (0.20 g, 0.27 mmol) in acetonitrile (5 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (100 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (1.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)-2-methylbenzoate hydrochloride (compound 14) (105 mg, yield 57%, white solid). 1 H NMR(400MHz,DMSO-d6)δ8.68(d,1H),8.34(m,4H),8.15(d,1H),7.76-7.67(m,3H),7.42(s,1H),6.13(s,2H),5.83(m,1H),4.72(m,1H),4.63 -4.56(m,1H),4.29-4.23(m,2H),4.05(m,1H),3.80-3.66(m,2H),3.55 (m,2H),2.64(s,3H),2.62(s,3H),2.50-2.44(m,1H),2.16(m,1H)ppm. LC-MS m / z(ESI)=636.4[M+H] + .

[0327] [Amended according to Rule 26, 31.07.2025] Example 15: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitoxy)propoxy)benzoate hydrochloride (compound 15)

[0328] Step 1: Preparation of (2,2-dimethyl-1,3-dioxolane-4-yl)methyl-4-methylbenzenesulfonate (compound 15b)

[0329] Under nitrogen protection, p-toluenesulfonyl chloride (26.0 g, 136.2 mmol) was added to (2,2-dimethyl-1,3-dioxolane-4-yl)methanol (compound 15a) (15.0 g, 113.5 mmol), pyridine (18.0 g, 227 mmol), and dichloromethane (200 mL) at 0 °C, and stirred at room temperature for 6 hours. The mixture was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (2,2-dimethyl-1,3-dioxolane-4-yl)methyl 4-methylbenzenesulfonate (compound 15b) (27 g, 83% yield, colorless oil). 1 H NMR (500MHz, CDCl3) δ7.83-7.73(d,2H),7.34(d,2H),4.30-4.21(m,1H),4.00(m,3H),3.75(m,1H),2.44(s,3H),1.33(s,3H),1.30(s,3H)ppm.

[0330] Step 2: Preparation of methyl 4-((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)benzoate (compound 15c)

[0331] Under nitrogen protection, methyl 4-hydroxybenzoate (13 g, 85.4 mmol), (2,2-dimethyl-1,3-dioxolane-4-yl)methyl 4-methylbenzenesulfonate (compound 15b) (29.4 g, 102.5 mmol), potassium carbonate (14.2 g, 102.5 mmol), and N,N-dimethylformamide (200 mL) were added sequentially to a reaction flask. The mixture was heated to 100 °C and stirred for 4 hours. The mixture was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 4-((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)benzoate (compound 15c) (16 g, 70% yield, colorless oil). LC-MS m / z (ESI) = 267.2 [M+H] + .

[0332] Step 3: Preparation of methyl 4-(2,3-dihydroxypropoxy)benzoate (compound 15d)

[0333] Under nitrogen protection, 10 mL of hydrochloric acid aqueous solution (60.1 mmol, 6N) was added dropwise to 100 mL of tetrahydrofuran containing methyl 4-((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)benzoate (compound 15c) (16 g, 60.1 mmol), and stirred at room temperature for 0.5 h. The mixture was concentrated under reduced pressure, and the pH was adjusted to 8 with sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (150 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then subjected to preparative separation under medium pressure to give methyl 4-(2,3-dihydroxypropoxy)benzoate (compound 15d) (13 g, 96% yield, colorless oil). LC-MS m / z (ESI) = 227.2 [M+H] + .

[0334] Step 4: Preparation of 4-(2,3-dihydroxypropoxy)benzoic acid (compound 15e)

[0335] Under nitrogen protection, methyl 4-(2,3-dihydroxypropoxy)benzoate (compound 15d) (6.0 g, 26.5 mmol), methanol (60 mL), water (20 mL), and sodium hydroxide (2.12 g, 53.00 mmol) were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (200 mL × 3), the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 4-(2,3-dihydroxypropoxy)benzoic acid (compound 15e) (5.3 g, 94% yield, white solid). 1 H NMR(400MHz,DMSO-d6)δ12.60(s,1H),7.89(d,2H),7.03(d,2H),5.04(s,1H), 4.74(s,1H),4.09(dd,1H),3.95(dd,1H),3.82(s,1H),3.46(m,2H)ppm; LC-MS m / z(ESI)=211.2[MH] - .

[0336] Step 5: Preparation of 4-(2,3-bis(nitoxy)propoxy)benzoic acid (compound 15f)

[0337] Under nitrogen protection, dichloromethane (50 mL), concentrated nitric acid (40 mL), and concentrated sulfuric acid (10 mL) were added sequentially to the reaction flask at 0 °C. The mixture was stirred for 30 minutes to prepare a mixed acid solution. A solution of 4-(2,3-dihydroxypropoxy)benzoic acid (compound 15e) (2.9 g, 13.7 mmol) in dichloromethane (50 mL) was added dropwise, and stirring continued for 0.5 hours after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (100 mL). The mixture was separated, extracted with dichloromethane (100 mL × 3), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 4-(2,3-bis(nitoxy)propoxy)benzoic acid (compound 15f) (0.3 g, yield 7%, white solid). LC-MS m / z (ESI) = 301.1 [MH] - .

[0338] Step 6: Preparation of chloromethyl 4-(2,3-bis(nitoxy)propoxy)benzoate (compound 15g)

[0339] Under nitrogen protection, 4-(2,3-bis(nitoxy)propoxy)benzoic acid (compound 15f) (0.30 g, 0.99 mmol), dichloromethane (10 mL), water (10 mL), sodium bicarbonate (0.42 g, 4.96 mmol), and tetrabutylammonium bisulfate (0.03 g, 0.10 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (0.25 g, 1.49 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparative separation to obtain chloromethyl 4-(2,3-bis(nitoxy)propoxy)benzoate (compound 15 g) (0.22 g, yield 63%, colorless oil).

[0340] Step 7: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)benzoate (compound 15h)

[0341] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.15 g, 0.37 mmol), methyltetrahydrofuran (5 mL), methyl 4-(2,3-bis(nitoxy)propoxy)benzoate (compound 15 g) (0.19 g, 0.55 mmol) and cesium carbonate (0.14 g, 0.44 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)benzoate (compound 15h) (0.15 g, yield 56%, white solid). LC-MS m / z (ESI) = 721.6 [M+H] + .

[0342] Step 8: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitoxy)propoxy)benzoate hydrochloride (compound 15)

[0343] p-Toluenesulfonic acid hydrate (0.16 g, 0.83 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitrooxy)propoxy)benzoate (compound 15h) (0.15 g, 0.21 mmol) in acetonitrile (5 mL) at room temperature and stirred for 0.5 h at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (1.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2,3-bis(nitoxy)propoxy)benzoate hydrochloride (compound 15) (38 mg, yield 28%, white solid). 1H NMR(500MHz,DMSO-d6)δ8.66(d,1H),8.42(s,3H),8.17(d,1H),7.93(d,2H),7.72-7.66(m,2H),7.09(d,2H),6.12(s,2H),5.83(m,1H),5.07(m, 1H),4.94(m,1H),4.52(m,1H),4.43(m,1H),4.07-4.01(m,1H),3.78-3. 67(m,2H),3.57-3.45(m,2H),2.61(s,3H),2.45(m,1H),2.16(m,1H)ppm. LC-MS m / z(ESI)=621.4[M+H] + .

[0344] [Amended according to Rule 26, 31.07.2025] Example 16: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitro)methyl)benzoate hydrochloride (compound 16)

[0345] Step 1: Preparation of 3-((nitro)methyl)benzoic acid (compound 16b)

[0346] Under nitrogen protection, 3-(bromomethyl)benzoic acid (compound 16a) (4.0 g, 18.6 mmol), silver nitrate (4.74 g, 27.90 mmol), and acetonitrile (40 mL) solution were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 12 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and the residue was separated by medium-pressure preparative chromatography to give compound 3-((nitrooxy)methyl)benzoic acid (compound 16b) (3.4 g, 93% yield, white solid). LC-MS m / z (ESI) = 196.1 [MH] - .

[0347] Step 2: Preparation of chloromethyl 3-((nitro)methyl)benzoate (compound 16c)

[0348] Under nitrogen protection, 3-((nitoxy)methyl)benzoic acid (compound 16b) (3.2 g, 16.2 mmol), dichloromethane (70 mL), water (70 mL), sodium bicarbonate (6.82 g, 81.20 mmol), and tetrabutylammonium bisulfate (0.55 g, 1.62 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (3.48 g, 21.10 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 3-((nitoxy)methyl)benzoate (compound 16c) (3.4 g, 85% yield, colorless oil). 1 H NMR (400MHz, CDCl3) δ8.11(m,2H),7.66(d,1H),7.59-7.46(m,1H),5.96(s,2H),5.48(s,2H)ppm.

[0349] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitoxy)benzoate methyl ester (compound 16d))

[0350] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), methyl 3-((nitoxy)methyl)benzoate chloromethyl ester (compound 16c) (0.30 g, 1.23 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitoxy)benzoate methyl ester (compound 16d)) (0.71 g, 94% yield, white solid). LC-MS m / z (ESI) = 617.5 [M+H] + .

[0351] Step 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitro)methyl)benzoate hydrochloride (compound 16)

[0352] At room temperature, p-toluenesulfonic acid hydrate (0.88 g, 4.60 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitoxy)benzoate methyl ester (compound 16d) (0.71 g, 1.15 mmol) in acetonitrile (10 mL), and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 with sodium bicarbonate solution. The aqueous phase was then treated with... Extracted with ethyl acetate (100 mL), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((nitro)methyl)benzoate hydrochloride (compound 16) (540 mg, yield 85%, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.66(d,1H),8.51(s,3H),8.19(d,1H),8.07(s,1H),8.01(d,1H),7.79(d,1H),7.72-7.67(m,2H),7.60( t,1H),6.17(s,2H),5.64(s,2H),4.04(s,1H),3.80-3.66(m,2H),3.56-3.45(m,2H),2.62(s,3H),2.44(m,1H),2.18(s,1H)ppm. LC-MS m / z(ESI)=517.3[M+H] + .

[0353] [Amended according to Rule 26, 31.07.2025] Example 17: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(1-(nitrooxy)ethyl)benzoate hydrochloride (compound 17)

[0354] Step 1: Preparation of 4-(1-(nitoxy)ethyl)benzoic acid (compound 17b)

[0355] Under nitrogen protection, 4-(1-bromoethyl)benzoic acid (compound 17a) (3.0 g, 13.1 mmol), silver nitrate (3.34 g, 19.65 mmol), and acetonitrile (30 mL) solution were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 12 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and the residue was separated by medium-pressure preparative chromatography to give compound 4-(1-(nitrooxy)ethyl)benzoic acid (compound 17b) (2.6 g, 94% yield, white solid). LC-MS m / z (ESI) = 210.1 [MH] - .

[0356] Step 2: Preparation of chloromethyl 4-(1-(nitoxy)ethyl)benzoate (compound 17c)

[0357] Under nitrogen protection, 4-(1-(nitoxy)ethyl)benzoic acid (compound 17b) (2.5 g, 11.8 mmol), dichloromethane (70 mL), water (70 mL), sodium bicarbonate (4.97 g, 59.20 mmol), and tetrabutylammonium bisulfate (0.40 g, 1.18 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (2.54 g, 15.40 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain chloromethyl 4-(1-(nitoxy)ethyl)benzoate (compound 17c) (2.0 g, yield 65%, colorless oil).

[0358] Step 3: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(1-(nitrooxy)ethyl)benzoate (compound 17d)

[0359] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), chloromethyl 4-(1-(nitoxy)ethyl)benzoate (compound 17c) (0.64 g, 2.45 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 1 hour. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(1-(nitrooxy)ethyl)benzoate (compound 17d) (0.71 g, 92% yield, white solid). LC-MS m / z (ESI) = 631.5 [M+H] + .

[0360] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(1-(nitrooxy)ethyl)benzoate hydrochloride (compound 17)

[0361] p-Toluenesulfonic acid hydrate (0.86 g, 4.50 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(1-(nitrooxy)ethyl)benzoate (compound 17d) (0.71 g, 1.13 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (100 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(1-(nitrooxy)ethyl)benzoate hydrochloride (compound 17) (538 mg, yield 84%, white solid). 1H NMR(500MHz,DMSO-d6)δ8.62(m,4H),8.20(m,1H),8.00(d,2H),7.69(m,2H),7.59(d,2H),6.21-6.11(m,3H),4.04(s,1H ),3.77(dd,1H),3.71(dd,1H),3.56-3.48(m,1H),3.35(d,1H),2.61(s,3H),2.44(m,1H),2.21(m,1H),1.56(d,3H)ppm. LC-MS m / z(ESI)=531.2[M+H] + .

[0362] [Amended according to Rule 26, 31.07.2025] Example 18: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethoxy)benzoate hydrochloride (compound 18)

[0363] Step 1: Preparation of methyl 4-(2-bromoethoxy)benzoate (compound 18b)

[0364] Under nitrogen protection, diisopropyl azodicarbonate (9.97 g, 49.29 mmol) was added dropwise to methyl 4-hydroxybenzoate (13a) (5.0 g, 32.9 mmol), 2-bromoethanol-1-ol (4.93 g, 39.40 mmol), triphenylphosphine (12.93 g, 49.29 mmol), and tetrahydrofuran (100 mL) at 0 °C, and stirred at room temperature for 3 hours. The mixture was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound methyl 4-(2-bromoethoxy)benzoate (compound 18b) (5.0 g, 59% yield, colorless oil). LC-MS m / z (ESI) = 260.1 [M+H] + .

[0365] Step 2: Preparation of 4-(2-bromoethoxy)benzoic acid (compound 18c)

[0366] Under nitrogen protection, methyl 4-(2-bromoethoxy)benzoate (compound 18b) (5.0 g, 19.3 mmol), tetrahydrofuran (50 mL), water (8 mL), and lithium hydroxide (1.85 g, 77.20 mmol) were added sequentially to the reaction flask. The mixture was stirred at 50 °C for 8 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (200 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 4-(2-bromoethoxy)benzoic acid (compound 18c) (3.1 g, yield 66%, white solid). LC-MS m / z (ESI) = 244.0 [MH] - .

[0367] Step 3: Preparation of 4-(2-(nitrooxy)ethoxy)benzoic acid (compound 18d)

[0368] Under nitrogen protection, 4-(2-bromoethoxy)benzoic acid (compound 18c) (2.00 g, 8.16 mmol), silver nitrate (2.77 g, 16.32 mmol), and acetonitrile (20 mL) solution were added sequentially to the reaction flask. The mixture was stirred at 75 °C for 16 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and the residue was separated by medium-pressure preparative chromatography to give compound 4-(2-(nitrooxy)ethoxy)benzoic acid (compound 18d) (1.7 g, 92% yield, white solid). LC-MS m / z (ESI) = 226.1 [MH] - .

[0369] Step 4: Preparation of chloromethyl 4-(2-(nitrooxy)ethoxy)benzoate (compound 18e)

[0370] Under nitrogen protection, 4-(2-(nitrooxy)ethoxy)benzoic acid (compound 18d) (1.70 g, 7.48 mmol), dichloromethane (30 mL), water (30 mL), sodium bicarbonate (3.14 g, 37.42 mmol), and tetrabutylammonium bisulfate (0.25 g, 0.75 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.60 g, 9.73 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 4-(2-(nitrooxy)ethoxy)benzoate (compound 18e) (1.4 g, yield 68%, colorless oil). 1H NMR (400MHz, CDCl3) δ8.05(d,2H),6.94(d,2H),5.94(s,2H),4.90-4.81(m,2H),4.37-4.24(m,2H)ppm.

[0371] Step 5: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethoxy)benzoate (compound 18f)

[0372] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), chloromethyl 4-(2-(nitrooxy)ethoxy)benzoate (compound 18e) (0.44 g, 1.60 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethoxy)benzoate (compound 18f) (0.72 g, 91% yield, white solid). LC-MS m / z (ESI) = 647.6 [M+H] + .

[0373] Step 6: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethoxy)benzoate hydrochloride (compound 18)

[0374] p-Toluenesulfonic acid hydrate (0.85 g, 4.45 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl4-(2-(nitrooxy)ethoxy)benzoate (compound 18f) (0.72 g, 1.11 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(2-(nitrooxy)ethoxy)benzoate hydrochloride (compound 18) (570 mg, yield 88%, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.65(d,1H),8.56(s,3H),8.19(d,1H),7.92(m,2H),7.69(m,2H),7.07(m,2H),6.12(s,2H),4.90( m,2H),4.40(m,2H),4.04(s,1H),3.76(m,1H),3.70(m,1H),3.58-3.47(m,2H),2.61(s,3H),2.44(m,1H),2.20(m,1H)ppm. LC-MS m / z(ESI)=547.2[M+H] + .

[0375] [Amended according to Rule 26, 31.07.2025] Example 19: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(3-(nitrooxy)propyl)benzoate hydrochloride (compound 19)

[0376] Step 1: Preparation of 4-(3-bromopropyl)benzoic acid (compound 19b)

[0377] Under nitrogen protection, methyl 4-(3-bromopropyl)benzoate (compound 19a) (2.00 g, 7.78 mmol), tetrahydrofuran (20 mL), water (4 mL), and lithium hydroxide (0.75 g, 31.11 mmol) were added sequentially to the reaction flask. The mixture was stirred at 50 °C for 8 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (50 mL × 2), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 4-(3-bromopropyl)benzoic acid (compound 19b) (1.4 g, 74% yield, white solid). LC-MS m / z (ESI) = 242.1 [MH] - .

[0378] Step 2: Preparation of 4-(3-(nitoxy)propyl)benzoic acid (compound 19c)

[0379] Under nitrogen protection, 4-(3-bromopropyl)benzoic acid (compound 19b) (1.40 g, 5.76 mmol), silver nitrate (1.47 g, 8.64 mmol), and acetonitrile (20 mL) solution were added sequentially to the reaction flask. The mixture was stirred at 75 °C for 16 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and the residue was separated by medium-pressure preparative chromatography to give compound 4-(3-(nitoxy)propyl)benzoic acid (compound 19c) (1.2 g, 92% yield, white solid). LC-MS m / z (ESI) = 224.2 [MH] - .

[0380] Step 3: Preparation of chloromethyl 4-(3-(nitoxy)propyl)benzoate (compound 19d)

[0381] Under nitrogen protection, 4-(3-(nitoxy)propyl)benzoic acid (compound 19c) (1.20 g, 5.33 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (2.24 g, 26.64 mmol), and tetrabutylammonium bisulfate (0.18 g, 0.53 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.05 g, 6.39 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 4-(3-(nitoxy)propyl)benzoate (compound 19d) (1.2 g, yield 82%, colorless oil). 1H NMR (400MHz, CDCl3) δ8.06(d,2H),7.33(d,2H),5.99(s,2H),4.49(t,2H),2.85(t,2H),2.17-2.03(m,2H)ppm.

[0382] Step 4: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(3-(nitrooxy)propyl)benzoate (compound 19e)

[0383] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), chloromethyl 4-(3-(nitoxy)propyl)benzoate (compound 19d) (0.40 g, 1.47 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(3-(nitrooxy)propyl)benzoate (compound 19e) (0.71 g, 90% yield, white solid). LC-MS m / z (ESI) = 645.5 [M+H] + .

[0384] Step 5: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl4-(3-(nitrooxy)propyl)benzoate hydrochloride (compound 19) hydrate (0.84 g, 4.41 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl4-(3-(nitrooxy)propyl)benzoate (compound 19e) (0.71 g, 1.10 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (80 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-(3-(nitoxy)propyl)benzoate hydrochloride (compound 19) (560 mg, yield 88%, white solid). 1 H NMR(400MHz,DMSO-d6)δ8.65(m,4H),8.21(d,1H),7.90(d,2H),7.74-7.65(m,2H),7.40(d,2H),6.15(s,2H),4.52(t,2H),4.06 (s,1H),3.83-3.66(m,2H),3.57-3.49(m,2H),2.81-2.71(m,2H),2.62(s,3H),2.46(m,1H),2.22(m,1H),2.07-1.92(m,2H)ppm. LC-MS m / z(ESI)=545.3[M+H] + .

[0385] [Amended according to Rule 26, 31.07.2025] Example 20: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-(nitrooxy)cyclohexane-1-carboxylic acid hydrochloride (compound 20)

[0386] Step 1: Preparation of (1S,4S)-4-(nitrooxy)cyclohexane-1-carboxylic acid (compound 20b)

[0387] Under nitrogen protection, acetic anhydride (32 mL) and nitric acid (8 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of ethyl acetate (50 mL) containing (1S,4S)-4-hydroxycyclohexane-1-carboxylic acid (compound 20a) (3.00 g, 20.81 mmol) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (100 mL). The mixture was extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (1S,4S)-4-(nitrooxy)cyclohexane-1-carboxylic acid (compound 20b) (2.0 g, yield 51%, white solid). LC-MS m / z (ESI) = 188.1 [MH] - .

[0388] Step 2: Preparation of (1S,4S)-4-(nitoxy)cyclohexane-1-carboxylic acid methyl ester (compound 20c)

[0389] Under nitrogen protection, (1S,4S)-4-(nitrooxy)cyclohexane-1-carboxylic acid (compound 20b) (2.00 g, 10.57 mmol), dichloromethane (50 mL), water (50 mL), sodium bicarbonate (4.44 g, 52.86 mmol), and tetrabutylammonium bisulfate (0.36 g, 1.06 mmol) were added dropwise to the reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated into liquid and liquid phases. The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain (1S,4S)-4-(nitrooxy)cyclohexane-1-carboxylic acid chloromethyl ester (compound 20c) (2.0 g, 80% yield, colorless oil).

[0390] Step 3: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-(nitrooxy)cyclohexane-1-carboxylate (compound 20d)

[0391] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), (1s,4s)-4-(nitoxy)cyclohexane-1-carboxylic acid chloromethyl ester (compound 20c) (0.44 g, 1.84 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-(nitrooxy)cyclohexane-1-carboxylate (compound 20d) (0.7 g, 93% yield, white solid). LC-MS m / z (ESI) = 609.5 [M+H] + .

[0392] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-(nitrooxy)cyclohexane-1-carboxylic acid hydrochloride (compound 20)

[0393] p-Toluenesulfonic acid hydrate (0.84 g, 4.41 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-(nitrooxy)cyclohexane-1-carboxylate (compound 20d) (0.70 g, 1.15 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (60 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-(nitrooxy)cyclohexane-1-carboxylic acid hydrochloride (compound 20) (530 mg, yield 85%, white solid). 1H NMR (400MHz, DMSO-d6) δ8.71-8.50(m,4H),8.20(d,1H),7.70(t,1H),7.57(s,1H),5.92(s,2H),5.16(s,1H),4.05(s,1H),3.82-3.66( m,2H),3.56-3.47(m,2H),2.59(s,3H),2.56(m,1H),2.45(m,1H),2.26-2.16(m,1H),1.85(m,2H),1.76(m,4H),1.69-1.57(m,2H)ppm. LC-MS m / z(ESI)=509.5[M+H] + .

[0394] [Amended according to Rule 26, 31.07.2025] Example 21: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-(nitrooxy)cyclohexane-1-carboxylic acid hydrochloride (compound 21)

[0395] Step 1: Preparation of (1R,4R)-4-(nitrooxy)cyclohexane-1-carboxylic acid (compound 21b)

[0396] Under nitrogen protection, acetic anhydride (32 mL) and nitric acid (8 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of ethyl acetate (50 mL) containing (1R,4R)-4-hydroxycyclohexane-1-carboxylic acid (compound 21a) (3.00 g, 20.81 mmol) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (100 mL). The mixture was extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (1R,4R)-4-(nitrooxy)cyclohexane-1-carboxylic acid (compound 21b) (3.2 g, yield 81%, white solid). LC-MS m / z (ESI) = 188.1 [MH] - .

[0397] Step 2: Preparation of (1R,4R)-4-(nitoxy)cyclohexane-1-carboxylic acid methyl ester (compound 21c)

[0398] Under nitrogen protection, (1R,4R)-4-(nitrooxy)cyclohexane-1-carboxylic acid (compound 21b) (3.20 g, 16.92 mmol), dichloromethane (70 mL), water (70 mL), sodium bicarbonate (7.11 g, 84.58 mmol), and tetrabutylammonium bisulfate (0.57 g, 1.69 mmol) were added dropwise to the reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated into liquid and liquid phases. The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (1R,4R)-4-(nitrooxy)cyclohexane-1-carboxylic acid chloromethyl ester (compound 21c) (3.5 g, yield 87%, colorless oil).

[0399] Step 3: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-(nitrooxy)cyclohexane-1-carboxylate (compound 21d)

[0400] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), (1r,4r)-4-(nitoxy)cyclohexane-1-carboxylic acid chloromethyl ester (compound 21c) (0.44 g, 1.84 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-(nitrooxy)cyclohexane-1-carboxylate (compound 21d) (0.69 g, 92% yield, white solid). LC-MS m / z (ESI) = 609.5 [M+H] + .

[0401] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-(nitrooxy)cyclohexane-1-carboxylic acid hydrochloride (compound 21)

[0402] p-Toluenesulfonic acid hydrate (0.84 g, 4.41 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-(nitrooxy)cyclohexane-1-carboxylate (compound 21d) (0.70 g, 1.15 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (60 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,4S)-4-(nitrooxy)cyclohexane-1-carboxylic acid hydrochloride (compound 21) (505 mg, yield 81%, white solid). 1 H NMR (400MHz, DMSO-d6) δ8.69-8.51(m,4H),8.20(d,1H),7.71(t,1H),7.57(s,1H),5.91(s,2H),5.06-4.93(m,1H),4.06(s, 1H),3.75(m,2H),3.58-3.48(m,2H),2.60(s,3H),2.50-2.40(m,2H),2.21(m,1H),2.09-1.92(m,4H),1.61-1.39(m,4H)ppm. LC-MS m / z(ESI)=509.3[M+H] + .

[0403] [Amended according to Rule 26, 31.07.2025] Example 22: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-((nitrooxy)methyl)1-cyclobutane hydrochloride (compound 22)

[0404] Step 1: Preparation of (1R,3R)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 22b)

[0405] Under nitrogen protection, acetic anhydride (10 mL) and nitric acid (3 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of (1R,3R)-3-(hydroxymethyl)cyclobutane-1-carboxylic acid (compound 22a) (1.50 g, 11.53 mmol) in dichloromethane (20 mL) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (50 mL). The mixture was extracted with dichloromethane (50 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (1R,3R)-3-((nitrooxy)methyl)cyclobutane-1-carboxylic acid (compound 22b) (2.0 g, 99% yield, colorless oil). LC-MS m / z (ESI) = 174.1 [MH] - .

[0406] Step 2: Preparation of (1R,3R)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid chloromethyl ester (compound 22c)

[0407] Under nitrogen protection, (1R,3R)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 22b) (2.00 g, 11.42 mmol), dichloromethane (50 mL), water (50 mL), sodium bicarbonate (4.8 g, 57.1 mmol), and tetrabutylammonium bisulfate (0.39 g, 1.14 mmol) were added dropwise to the reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated into liquid and liquid phases. The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain (1R,3R)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid chloromethyl ester (compound 22c) (1.4 g, yield 55%, colorless oil). 1 H NMR (500MHz, CDCl3) δ5.72(s,2H),4.48(d,2H),3.26-3.14(m,1H),2.90-2.74(m,1H),2.55-2.43(m,2H),2.17(m,2H)ppm.

[0408] Step 3: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-((nitrooxy)methyl)cyclobutane-1-carboxylic acid ester (compound 22d)

[0409] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), (1R,3R)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid chloromethyl ester (compound 22c) (0.41 g, 1.84 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-((nitrooxy)methyl)cyclobutane-1-carboxylic acid ester (compound 22d) (0.71 g, 96% yield, white solid). LC-MS m / z (ESI) = 595.4 [M+H] + .

[0410] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-((nitrooxy)methyl)1-cyclobutane hydrochloride (compound 22)

[0411] p-Toluenesulfonic acid hydrate (0.902 g, 4.742 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-((nitrooxy)methyl)cyclobutane-1-carboxylic acid ester (compound 22d) (0.705 g, 1.186 mmol)) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (60 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-((nitro)methyl)1-cyclobutane hydrochloride (compound 22) (590 mg, 94% yield, white solid). 1H NMR(500MHz,DMSO-d6)δ8.63(d,1H),8.54(s,3H),8.18(d,1H),7.69(t,1H),7.56(s,1H),5.91(s,2H),4.58(d,2H),4.03(m,1H),3.76(dd, 1H),3.70(dd,1H),3.57-3.47(m,2H),3.30-3.22(m,1H),2.63(m,1H),2.58(s,3H),2.44(m,1H),2.31(m,2H),2.19(m,1H),2.07(m,2H)ppm. LC-MS m / z(ESI)=495.4[M+H] + .

[0412] [Amended according to Rule 26, 31.07.2025] Example 23: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((nitro)methyl)benzoate hydrochloride (compound 23)

[0413] Step 1: Preparation of 4-((nitoxy)methyl)benzoic acid (compound 23b)

[0414] Under nitrogen protection, 4-(bromomethyl)benzoic acid (compound 23a) (1.00 g, 4.65 mmol), silver nitrate (0.95 g, 5.58 mmol), and acetonitrile (10 mL) solution were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 12 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and the residue was separated by medium-pressure preparative chromatography to give compound 4-((nitrooxy)methyl)benzoic acid (compound 23b) (0.92 g, 99% yield, white solid). LC-MS m / z (ESI) = 196.1 [MH] - .

[0415] Step 2: Preparation of chloromethyl 4-(nitrooxy)methyl)benzoate (compound 23c)

[0416] Under nitrogen protection, 4-((nitoxy)methyl)benzoic acid (compound 23b) (0.92 g, 4.67 mmol), dichloromethane (30 mL), water (30 mL), sodium bicarbonate (1.96 g, 23.33 mmol), and tetrabutylammonium bisulfate (0.16 g, 0.47 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethylsulfonyl chloride (0.92 g, 5.60 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparative separation to obtain chloromethyl 4-(nitoxy)methyl)benzoate (compound 23c) (1.0 g, yield 87%, colorless oil). 1 H NMR (400MHz, CDCl3) δ8.12(d,2H),7.50(d,2H),5.96(s,2H),5.49(s,2H)ppm.

[0417] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((nitoxy)benzoate methyl ester (compound 23d))

[0418] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), chloromethyl 4-(nitrooxy)methyl)benzoate (compound 23c) (0.45 g, 1.84 mmol), and cesium carbonate (0.44 g, 1.35 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((nitoxy)benzoate methyl ester (compound 23d)) (0.5 g, yield 66%, white solid). LC-MS m / z (ESI) = 617.5 [M+H] + .

[0419] Step 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((nitro)methyl)benzoate hydrochloride (compound 23)

[0420] At room temperature, p-toluenesulfonic acid hydrate (0.62 g, 3.24 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((nitoxy)benzoate methyl ester (compound 23d)) (0.50 g, 0.81 mmol) in acetonitrile (10 mL), and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 with sodium bicarbonate solution. The aqueous phase was then treated with... Extracted with ethyl acetate (100 mL), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((nitro)methyl)benzoate hydrochloride (compound 23) (310 mg, yield 69%, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.68-8.64(m,1H),8.54(s,3H),8.19(d,1H),8.01(d,2H),7.69(m,2H),7.60(m,2H),6.16(s,2 H),5.66(s,2H),4.03(m,1H),3.77(m,1H),3.74-3.68(m,1H),3.52(m,2H),2.61(s,3H),2.44(m,1H),2.20(m,1H)ppm. LC-MS m / z(ESI)=517.5[M+H] + .

[0421] Example 24: Preparation of methyl 4-(5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1-hydroxy-3-(nitrooxy)propane)-2-yl)oxy)-2-benzoate hydrochloride (compound 24)

[0422] Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1-hydroxy-3-(nitrooxy)propane)-2-yl)oxy)-2-benzoate hydrochloride (compound 24)

[0423] Palladium on carbon (0.01 g) was added to a solution of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1,3-bis(nitrooxy)prop-2-yl)oxy)-2-methylbenzoate hydrochloride (compound 13) (0.13 g, 0.19 mmol), triethylamine (0.02 g, 0.19 mmol), and ethyl acetate (5 mL) at room temperature and stirred for 2 hours at room temperature. The mixture was filtered and concentrated under reduced pressure. The residue was then separated under high pressure. The product was added to an ethyl acetate hydrochloride solution (0.5 mL, 4 N) and concentrated under reduced pressure to obtain methyl 5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-4-((1-hydroxy-3-(nitrooxy)propane)-2-yl)oxy)-2-benzoate hydrochloride (compound 24) (19 mg, yield 15%, white solid). 1 H NMR(400MHz,DMSO-d6)δ8.67(d,1H),8.40(s,3H),8.15(d,1H),7.80(d,1H),7.70-7.65(m,2H),7.01(d,1H),6.90(m,1 H),4.83(dd,2H),4.05(m,1H),3.90-3.67(m,6H),3.53(m,4H),2.63(s,3H),2.53(s,3H),2.46(m,1H),2.19(m,1H)ppm. LC-MS m / z(ESI)=591.3[M+H] + .

[0424] [Amended according to Rule 26, 31.07.2025] Example 25: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-hydroxy-2-methyl-2-((nitro)methyl)propionate hydrochloride (compound 25)

[0425] Step 1: Preparation of 3-hydroxy-2-methyl-2-((nitro)methyl)propionic acid (compound 25b)

[0426] Under nitrogen protection, acetic anhydride (20 mL) and nitric acid (3 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of ethyl acetate (50 mL) containing 3-hydroxy-2-hydroxymethyl-2-methylpropionic acid (compound 25a) (5.00 g, 37.28 mmol) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (50 mL). The mixture was extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 3-hydroxy-2-methyl-2-((nitoxy)methyl)propionic acid (compound 25b) (2.2 g, yield 33%, white solid). LC-MS m / z (ESI) = 178.1 [MH] - .

[0427] Step 2: Preparation of 3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitrooxy)methyl)propionic acid (compound 25c)

[0428] Under nitrogen protection, 3-hydroxy-2-methyl-2-((nitro)methyl)propionic acid (compound 25b) (2.20 g, 12.28 mmol), dichloromethane (30 mL), imidazole (2.51 g, 36.85 mmol), and tert-butyldimethylchlorosilane (4.63 g, 30.70 mmol) were added sequentially to the reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 0.5 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and methanol (30 mL) and potassium carbonate (2.55 g, 1 mmol) were added. 8.42 mmol), stirred at room temperature for 1 hour; concentrated under reduced pressure, pH adjusted to 4 with hydrochloric acid aqueous solution (1N), extracted with ethyl acetate (100 mL × 3), the organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitrooxy)methyl)propionic acid (compound 25c) (1.4 g, yield 39%, colorless oil). LC-MS m / z (ESI) = 292.3 [MH] - .

[0429] Step 3: Preparation of chloromethyl 3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitoxy)methyl)propionate (compound 25d)

[0430] Under nitrogen protection, at 0°C, 3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitrooxy)methyl)propionic acid (compound 25c) (1.40 g, 4.77 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (2.00 g, 23.86 mmol), and tetrabutylammonium bisulfate (0.16 g, 0.48 mmol) were added sequentially to the reaction flask. Chloromethylsulfonyl chloride (0.94 g) was then added dropwise. g, 5.73 mmol), after addition, return to room temperature and stir for 2 hours; TLC monitoring showed complete reaction; separation, organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, filtrate concentrated under reduced pressure, residue was separated under medium pressure to give compound 3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitoxy)methyl)propionate chloromethyl ester (compound 25d) (0.61 g, yield 37%, colorless oil).

[0431] Step 4: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitrooxy)methyl)propionate (compound 25e)

[0432] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (0.30 g, 0.73 mmol), methyltetrahydrofuran (10 mL), methyl 3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitoxy)methyl)propionate (compound 25d) (0.38 g, 1.10 mmol) and cesium carbonate (0.24 g, 0.74 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitrooxy)methyl)propionate (compound 25e) (0.42 g, yield 81%, white solid). LC-MS m / z (ESI) = 713.7 [M+H] + .

[0433] Step 5: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-hydroxy-2-methyl-2-((nitro)methyl)propionate hydrochloride (compound 25)

[0434] p-Toluenesulfonic acid hydrate (0.85 g, 4.49 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((tert-butyldimethylsilyl)oxy)-2-methyl-2-((nitrooxy)methyl)propionate (compound 25e) (0.40 g, 0.56 mmol) in acetonitrile (10 mL) at room temperature and stirred for 2 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (100 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-hydroxy-2-methyl-2-((nitro)methyl)propionate hydrochloride (compound 25) (170 mg, yield 57%, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.30(s,3H),8.19-8.10(m,1H),7.69(t,1H),7.52(s,1H),5.93(s,2H),5.21(m,1H),4. 71(d,1H),4.57(d,1H),4.04(m,1H),3.72(m,2H),3.60-3.45(m,4H),2.59(s,3H),2.45(m,1H),2.14(m,1H),1.13(s,3H)ppm. LC-MS m / z(ESI)=499.2[M+H] + .

[0435] [Amended according to Rule 26, 31.07.2025] Example 26: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-(hydroxymethyl)-3-((nitrooxy)methyl)1-cyclobutane hydrochloride (compound 26)

[0436] Step 1: Preparation of 3-(hydroxymethyl)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 26b)

[0437] Under nitrogen protection, acetic anhydride (20 mL) and nitric acid (5 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of ethyl acetate (20 mL) of 3,3-bis(hydroxymethyl)cyclobutane-1-carboxylic acid (compound 26a) (1.40 g, 8.74 mmol) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched with water (50 mL). The mixture was extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 3-(hydroxymethyl)-3-((nitrooxy)methyl)cyclobutane-1-carboxylic acid (compound 26b) (0.14 g, yield 8%, white solid). LC-MS m / z (ESI) = 204.1 [MH] - .

[0438] Step 2: Preparation of 3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 26c)

[0439] Under nitrogen protection, 3-(hydroxymethyl)-3-((nitrooxy)methyl)cyclobutane-1-carboxylic acid (compound 26b) (0.150 g, 0.731 mmol), dichloromethane (10 mL), imidazole (0.15 g, 2.19 mmol), and tert-butyldimethylchlorosilane (0.28 g, 1.83 mmol) were added sequentially to the reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 0.5 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and methanol (10 mL) and potassium carbonate (0.15 g, 0.731 mmol) were added. 1.10 mmol), stirred at room temperature for 1 hour; concentrated under reduced pressure, pH adjusted to 4 with hydrochloric acid aqueous solution (1N), extracted with ethyl acetate (30 mL × 3), the organic phase washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, the filtrate concentrated under reduced pressure, and the residue subjected to preparative separation under medium pressure to give compound 3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid (compound 26c) (0.2 g, yield 87%, colorless oil). LC-MS m / z (ESI) = 318.3 [MH] - .

[0440] Step 3: Preparation of chloromethyl 3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitoxy)methyl)cyclobutane-1-carboxylate (compound 26d)

[0441] Under nitrogen protection, at 0°C, 3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitro)methyl)cyclobutane-1-carboxylic acid (compound 26c) (0.20 g, 0.63 mmol), dichloromethane (10 mL), water (10 mL), sodium bicarbonate (0.26 g, 3.13 mmol), and tetrabutylammonium bisulfate (0.04 g, 0.13 mmol) were added sequentially to the reaction flask. Chloromethylsulfonyl chloride (0.21 g) was then added dropwise. 1.25 mmol), after addition, return to room temperature and stir for 2 hours; TLC monitoring showed complete reaction; separation, organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, filtrate concentrated under reduced pressure, residue was separated under medium pressure to give compound 3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitoxy)methyl)cyclobutane-1-carboxylic acid chloromethyl ester (compound 26d) (0.08 g, yield 35%, colorless oil).

[0442] Step 4: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitro)methyl)cyclobutane-1-carboxylic acid ester (compound 26e)

[0443] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (0.080 g, 0.196 mmol), methyltetrahydrofuran (5 mL), 3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitoxy)methyl)cyclobutane-1-carboxylate chloromethyl ester (compound 26d) (0.072 g, 0.196 mmol) and cesium carbonate (0.07 g, 0.22 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitro)methyl)cyclobutane-1-carboxylic acid ester (compound 26e) (0.078 g, yield 52%, white solid). LC-MS m / z (ESI) = 739.6 [M+H] + .

[0444] Step 5: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(hydroxymethyl)-3-((nitrooxy)methyl)1-cyclobutane hydrochloride (compound 26)

[0445] At room temperature, p-toluenesulfonic acid hydrate (0.081 g, 0.424 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-(((tert-butyldimethylsilyl)oxy)methyl)-3-((nitro)methyl)cyclobutane-1-carboxylic acid ester (compound 26e) (0.078 g, 0.106 mmol) in acetonitrile (5 mL), and stirred at room temperature for 2 hours. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (1.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-(hydroxymethyl)-3-((nitrooxy)methyl)1-cyclobutane hydrochloride (compound 26) (36 mg, yield 61%, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.29(s,3H),8.13(m,1H),7.69(t,1H),7.56(s,1H),5.91(s,2H),4.58(s,1H),4.46(s,2H),4 .04(s,1H),3.77-3.65(m,2H),3.54(m,1H),3.48-3.41(m,2H),3.20(m,1H),2.58(s,3H),2.49-2.41(m,2H),2.18-1.99(m,5H)ppm. LC-MS m / z(ESI)=525.1[M+H] + .

[0446] [Revised according to Rule 26, 31.07.2025] Example 27: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-6-((nitro)methyl)spiro[3.3]heptane-2-carboxylic acid hydrochloride (compound 27)

[0447] Step 1: Preparation of methyl 6-(hydroxymethyl)spiro[3.3]heptane-2-carboxylate (compound 27b)

[0448] Under nitrogen protection, 6-(methoxycarbonyl)spiro[3.3]heptane-2-carboxylic acid (compound 27a) (2.4 g, 12.1 mmol), tetrahydrofuran (20 mL), and boranetetrahydrofuran solution (13.3 mL, 13.3 mmol, 1.0 N) were added sequentially to the reaction flask at 0 °C. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. The reaction was quenched dropwise with ice water, extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain methyl 6-(hydroxymethyl)spiro[3.3]heptane-2-carboxylic acid (compound 27b) (2.2 g, 99% yield, colorless oil). LC-MS m / z (ESI) = 185.2 [M+H] + .

[0449] Step 2: Preparation of 6-(hydroxymethyl)spiro[3.3]heptane-2-carboxylic acid (compound 27c)

[0450] Under nitrogen protection, methyl 6-(hydroxymethyl)spiro[3.3]heptane-2-carboxylic acid (compound 27b) (2.2 g, 11.9 mmol), tetrahydrofuran (20 mL), water (5 mL), and sodium hydroxide (0.72 g, 11.91 mmol) were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (100 mL × 3), the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 6-(hydroxymethyl)spiro[3.3]heptane-2-carboxylic acid (compound 27c) (1.8 g, yield 89%, white solid). LC-MS m / z (ESI) = 169.2 [MH] - .

[0451] Step 3: Preparation of 6-((nitoxy)methyl)spiro[3.3]heptane-2-carboxylic acid (compound 27d)

[0452] Under nitrogen protection, acetic anhydride (40 mL) and concentrated nitric acid (10 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 minutes to prepare a mixed acid solution. A solution of 6-(hydroxymethyl)spiro[3.3]heptane-2-carboxylic acid (compound 27c) (1.80 g, 10.58 mmol) in ethyl acetate (50 mL) was added dropwise, and stirring continued for 0.5 hours after the addition was complete. The reaction was monitored by TLC until complete. The reaction was quenched with water (100 mL), and the mixture was separated. The extract was obtained by extraction with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to medium-pressure preparative separation to obtain compound 6-((nitrooxy)methyl)spiro[3.3]heptane-2-carboxylic acid (27d) (0.8 g, yield 35%, colorless oil). LC-MS m / z (ESI) = 214.2 [MH]. - .

[0453] Step 4: Preparation of chloromethyl 6-((nitoxy)methyl)spiro[3.3]heptane-2-carboxylate (compound 27e)

[0454] Under nitrogen protection, 6-((nitoxy)methyl)spiro[3.3]heptane-2-carboxylic acid (compound 27d) (0.80 g, 3.72 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (1.56 g, 18.59 mmol), and tetrabutylammonium bisulfate (0.25 g, 0.74 mmol) were added dropwise to the reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain chloromethyl 6-((nitoxy)methyl)spiro[3.3]heptane-2-carboxylic acid (compound 27e) (0.9 g, yield 92%, colorless oil).

[0455] Step 5: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-6-((nitrooxy)methyl)spiro[3.3]heptane-2-carboxylic acid ester (compound 27f)

[0456] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), chloromethyl 6-((nitoxy)methyl)spiro[3.3]heptane-2-carboxylate (compound 27e) (0.39 g, 1.47 mmol) and cesium carbonate (0.44 g, 1.35 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-6-((nitrooxy)methyl)spiro[3.3]heptane-2-carboxylic acid ester (compound 27f) (0.71 g, 91% yield, white solid). LC-MS m / z (ESI) = 635.7 [M+H] + .

[0457] Step 6: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-6-((nitro)methyl)spiro[3.3]heptane-2-carboxylic acid hydrochloride (compound 27)

[0458] p-Toluenesulfonic acid hydrate (0.85 g, 4.48 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-6-((nitrooxy)methyl)spiro[3.3]heptane-2-carboxylic acid ester (compound 27f) (0.71 g, 1.12 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. Concentrate under reduced pressure, add sodium bicarbonate solution to adjust pH=8, extract the aqueous phase with ethyl acetate (60 mL), combine the organic phases and dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the residue, prepare and separate under medium pressure, add the product to ethyl acetate hydrochloride solution (2.0 mL, 4 N), concentrate under reduced pressure to obtain (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl6-((nitro)methyl)spiro[3.3]heptane-2-carboxylic acid hydrochloride (compound 27) (580 mg, yield 91%, white solid). 1H NMR(400MHz,DMSO-d6)δ8.65(dd,1H),8.56(s,3H),8.20(d,1H),7.70(t,1H),7.55(s,1H),5.89(s,2H),4.47(m,2H),4.05(m,1H),3 .81-3.67(m,2H),3.53(m,2H),3.06(m,1H),2.59(s,3H),2.50-2.41(m,2H),2.32-2.13(m,6H),2.06-2.00(m,1H),1.82(m,2H)ppm. LC-MS m / z(ESI)=535.2[M+H] + .

[0459] [Amended according to Rule 26, 31.07.2025] Example 28: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-1-((nitro)methyl)cyclopropane-1-carboxylate hydrochloride (compound 28)

[0460] Step 1: Preparation of 1-((nitoxy)methyl)cyclopropane-1-carboxylic acid (compound 28b)

[0461] Under nitrogen protection, acetic anhydride (30 mL) and nitric acid (10 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 50 min to prepare a mixed acid solution. 1-(hydroxymethyl)cyclopropane-1-carboxylic acid (compound 28a) (2.0 g, 17.2 mmol) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The reaction was quenched with water (100 mL), extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 1-((nitoxy)methyl)cyclopropane-1-carboxylic acid (compound 28b) (1.3 g, yield 47%, colorless oil). LC-MS m / z (ESI) = 160.1 [MH] - .

[0462] Step 2: Preparation of chloromethyl 1-((nitoxy)methyl)cyclopropane-1-carboxylate (compound 28c)

[0463] Under nitrogen protection, 1-((nitoxy)methyl)cyclopropane-1-carboxylic acid (compound 28b) (1.30 g, 8.07 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (3.39 g, 40.35 mmol), and tetrabutylammonium bisulfate (0.55 g, 1.61 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.73 g, 10.49 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 1-((nitoxy)methyl)cyclopropane-1-carboxylic acid (compound 28c) (1.6 g, 95% yield, colorless oil). 1 H NMR (500MHz, CDCl3) δ5.72(s,2H),4.64(s,2H),1.58-1.50(m,2H),1.14(m,2H)ppm.

[0464] Step 3: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-1-((nitro)-1-carboxylic acid methyl)cyclopropane ester (compound 28d)

[0465] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (20 mL), chloromethyl 1-((nitoxy)methyl)cyclopropane-1-carboxylate (compound 28c) (0.31 g, 1.47 mmol) and cesium carbonate (0.44 g, 1.35 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-1-((nitro)-1-carboxylic acid methyl)cyclopropane ester (compound 28d) (0.67 g, 94% yield, white solid). LC-MS m / z (ESI) = 581.5 [M+H] + .

[0466] Step 4: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-1-((nitro)methyl)cyclopropane-1-carboxylate hydrochloride (compound 28)

[0467] p-Toluenesulfonic acid hydrate (0.875 g, 4.600 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-1-((nitro)-1-carboxylic acid methyl)cyclopropane ester (compound 28d) (0.67 g, 1.15 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (100 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give compound (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-1-((nitro)methyl)cyclopropane-1-carboxylate hydrochloride (compound 28) (540 mg, yield 91%, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.38(s,3H),8.15(d,1H),7.69(t,1H),7.53(s,1H),5.90(s,2H),4.69(s,2H),4.04(m, 1H),3.79-3.67(m,2H),3.57-3.46(m,2H),2.58(s,3H),2.47-2.41(m,1H),2.17(m,1H),1.31(m,2H),1.23-1.19(m,2H)ppm. LC-MS m / z(ESI)=481.2[M+H] + .

[0468] [Amended according to Rule 26, 31.07.2025] Example 29: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2,2-dimethyl-3-(3-(nitrooxy)-2-((nitrooxy)methyl)propoxy)propionate hydrochloride (compound 29)

[0469] Step 1: Preparation of methyl 4-methylbenzenesulfonic acid (2,2-dimethyl-1,3-dioxane-5-yl) ester (compound 29b)

[0470] Under nitrogen protection, p-toluenesulfonyl chloride (11.5 g, 60.2 mmol) was added to (2,2-dimethyl-1,3-dioxane-5-yl)methanol (compound 29a) (8.0 g, 54.72 mmol), triethylamine (6.64 g, 65.67 mmol), and dichloromethane (100 mL) at 0 °C, and stirred at room temperature for 12 hours. The mixture was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 4-methylbenzenesulfonic acid (2,2-dimethyl-1,3-dioxane-5-yl) methyl ester (29b) (14 g, 85% yield, colorless oil).

[0471] Step 2: Preparation of methyl 3-((2,2-dimethyl-1,3-dioxane-5-yl)methoxy)-2,2-dimethylpropionate (compound 29c)

[0472] Under nitrogen protection, sodium hydride (1.09 g, 27.20 mmol, 60%) was added to a solution of methyl 3-hydroxy-2,2-dimethylpropionate (3.0 g, 22.7 mmol) in N,N-dimethylformamide (40 mL) at 0 °C and stirred for 30 minutes; then methyl 4-methylbenzenesulfonic acid (2,2-dimethyl-1,3-dioxane-5-yl) (compound 29b) (8.18 g, 27.24 mmol) was added, and the mixture was heated to 60 °C and stirred for 2 hours. The temperature was lowered to 0°C, and the reaction was quenched by adding saturated ammonium chloride solution (100 mL). Extraction was performed with ethyl acetate (100 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then subjected to preparative separation under medium pressure to give compound 3-((2,2-dimethyl-1,3-dioxane-5-yl)methoxy)-2,2-dimethylpropionate (compound 29c) (1.7 g, 29% yield, colorless oil). LC-MS m / z (ESI) = 261.3 [M + H] + .

[0473] Step 3: Preparation of 3-((2,2-dimethyl-1,3-dioxane-5-yl)methoxy)-2,2-dimethylpropionic acid (compound 29d)

[0474] Under nitrogen protection, an aqueous solution of sodium hydroxide (0.39 g, 9.80 mmol, 1 N) was added dropwise to methyl 3-((2,2-dimethyl-1,3-dioxane-5-yl)methoxy)-2,2-dimethylpropionic acid (compound 29c) (1.70 g, 6.53 mmol) in tetrahydrofuran (30 mL), and stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the pH was adjusted to 4 with aqueous hydrochloric acid. The aqueous phase was extracted with ethyl acetate (150 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 3-((2,2-dimethyl-1,3-dioxane-5-yl)methoxy)-2,2-dimethylpropionic acid (compound 29d) (1.2 g, 75% yield, colorless oil). LC-MS m / z (ESI) = 245.2 [MH] - .

[0475] Step 4: Preparation of 3-(3-hydroxy-2-(hydroxymethyl)propoxy)-2,2-dimethylpropionic acid (compound 29e)

[0476] Under nitrogen protection, 3-((2,2-dimethyl-1,3-dioxane-5-yl)methoxy)-2,2-dimethylpropionic acid (compound 29d) (1.20 g, 4.87 mmol), tetrahydrofuran (20 mL), and aqueous hydrochloric acid solution (0.18 g, 4.87 mmol, 1 N) were added sequentially to the reaction flask. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. The mixture was extracted with ethyl acetate (100 mL × 3), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 3-(3-hydroxy-2-(hydroxymethyl)propoxy)-2,2-dimethylpropionic acid (compound 29e) (1.0 g, 99% yield, colorless oil). LC-MS m / z (ESI) = 205.2 [MH] - .

[0477] Step 5: Preparation of 2,2-dimethyl-3-(3-(nitoxy)-2-((nitoxy)methyl)propoxy)propionic acid (compound 29f)

[0478] Under nitrogen protection, acetic anhydride (20 mL) and concentrated nitric acid (5 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 minutes to prepare a mixed acid solution. A solution of ethyl acetate (20 mL) of 3-(3-hydroxy-2-(hydroxymethyl)propoxy)-2,2-dimethylpropionic acid (compound 29e) (1.00 g, 4.85 mmol) was added dropwise, and the mixture was stirred for 0.5 hours after the addition was complete. The reaction was monitored by TLC until it was complete. The mixture was cooled to 0 °C, and the reaction was quenched with water (50 mL). The mixture was separated, extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain compound 2,2-dimethyl-3-(3-(nitoxy)-2-((nitoxy)methyl)propoxy)propionic acid (compound 29f) (0.31 g, yield 22%, colorless oil). LC-MS m / z (ESI) = 295.1 [MH] - .

[0479] Step 6: Preparation of chloromethyl 2,2-dimethyl-3-(3-(nitoxy)-2-((nitoxy)methyl)propoxy)propionate (compound 29g)

[0480] Under nitrogen protection, at 0°C, 2,2-dimethyl-3-(3-(nitoxy)-2-((nitoxy)methyl)propoxy)propionic acid (compound 29f) (0.31 g, 1.05 mmol), dichloromethane (10 mL), water (10 mL), sodium bicarbonate (0.44 g, 5.23 mmol), and tetrabutylammonium bisulfate (0.07 g, 0.21 mmol) were added sequentially to the reaction flask, followed by dropwise addition of chloromethylsulfonyl chloride (0.22 g). 1.36 mmol), after the addition was complete, the mixture was returned to room temperature and stirred for 1 hour; the reaction was monitored by TLC until complete; the mixture was separated, the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by medium pressure to give methyl chloropropionate 2,2-dimethyl-3-(3-(nitoxy)-2-((nitoxy)methyl)propoxy)propionate (compound 29 g) (0.25 g, yield 69%, colorless oil).

[0481] Step 7: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2,2-dimethyl-3-(3-(nitrooxy)-2-((nitrooxy)methyl)propoxy)propionate (compound 29h)

[0482] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.13 g, 0.32 mmol), methyltetrahydrofuran (5 mL), methyl 2,2-dimethyl-3-(3-(nitrooxy)-2-((nitrooxy)methyl)propoxy)chloromethyl propionate (compound 29 g) (0.16 g, 0.48 mmol) and cesium carbonate (0.11 g, 0.35 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2,2-dimethyl-3-(3-(nitrooxy)-2-((nitrooxy)methyl)propoxy)propionate (compound 29h) (0.16 g, 70% yield, white solid). LC-MS m / z (ESI) = 716.6 [M+H] + .

[0483] Step 8: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2,2-dimethyl-3-(3-(nitrooxy)-2-((nitrooxy)methyl)propoxy)propionate hydrochloride (compound 29)

[0484] At room temperature, p-toluenesulfonic acid hydrate (0.16 g, 0.84 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2,2-dimethyl-3-(3-(nitrooxy)-2-((nitrooxy)methyl)propoxy)propionate (compound 29h) (0.15 g, 0.21 mmol) in acetonitrile (5 mL), and stirred at room temperature for 0.5 hours. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (1.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2,2-dimethyl-3-(3-(nitro)-2-((nitro)methyl)propoxy)propionate hydrochloride (compound 29) (83 mg, yield 61%, white solid). 1H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.26(s,3H),8.12(d,1H),7.67(t,1H),7.53(s,1H),5.90(s,2H),4.51-4.42(m,4H),3.77 -3.66(m,2H),3.55-3.45(m,2H),3.41-3.38(m,4H),2.59(s,3H),2.47-2.37(m,2H),2.12(m,1H),1.35(m,1H),1.10(s,6H)ppm. LC-MS m / z(ESI)=616.3[M+H] + .

[0485] [Amended according to Rule 26, 31.07.2025] Example 30: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2-hydroxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionate hydrochloride (compound 30)

[0486] Step 1: Preparation of methyl 3-(allyloxy)-2,2-dimethylpropionate (compound 30b)

[0487] Under nitrogen protection, methyl 3-hydroxy-2,2-dimethylpropionate (compound 30a) (10.00 g, 75.67 mmol), tetrahydrofuran (100 mL), and sodium hydride (3.00 g, 75.67 mmol, 60%) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 minutes. Allyl bromide (9.15 g, 75.67 mmol) was added dropwise, and the reaction was stirred at room temperature for 1 hour. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the mixture was quenched by adding saturated ammonium chloride solution (100 mL). The aqueous phase was extracted with ethyl acetate (100 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then subjected to preparative separation under medium pressure to give compound methyl 3-(allyloxy)-2,2-dimethylpropionate (compound 30b) (8.4 g, yield 64%, colorless oil). LC-MS m / z (ESI) = 173.2 [M+H] + .

[0488] Step 2: Preparation of methyl 3-(2,3-dihydroxypropoxy)-2,2-dimethylpropionate (30c)

[0489] Under nitrogen protection, tert-butanol (300 mL), purified water (300 mL), and AD-mix-α (50 g) were added sequentially to the reaction flask. The mixture was stirred at room temperature for ten minutes, then cooled to 0 °C. Methyl 3-(allyloxy)-2,2-dimethylpropionate (compound 30b) (5.00 g, 29.03 mmol) was added dropwise, and the reaction was stirred for 4 hours. The reaction was monitored by TLC until complete. The reaction was quenched by adding sodium sulfite aqueous solution (100 mL). The aqueous phase was extracted with ethyl acetate (200 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then subjected to preparative separation under medium pressure to obtain compound 3-(2,3-dihydroxypropoxy)-2,2-dimethylpropionate (compound 30c) (4.1 g, yield 68%, colorless oil). LC-MS m / z (ESI) = 207.2 [M+H] + .

[0490] Step 3: Preparation of methyl 3-(3-((tert-butyldimethylsilyl)oxy)-2-hydroxypropoxy)-2,2-dimethylpropionate (compound 30d)

[0491] Under nitrogen protection, methyl 3-(2,3-dihydroxypropoxy)-2,2-dimethylpropionate (compound 30c) (4.0 g, 19.4 mmol), dichloromethane (60 mL), imidazole (1.58 g, 23.27 mmol), and tert-butyldimethylchlorosilane (2.92 g, 19.40 mmol) were added sequentially to the reaction flask at 0 °C. After the addition was complete, the mixture was returned to room temperature and stirred for 0.5 hours. The reaction was monitored by TLC until complete. The mixture was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparative separation to obtain methyl 3-(3-((tert-butyldimethylsilyl)oxy)-2-hydroxypropoxy)-2,2-dimethylpropionate (compound 30d) (4.4 g, 71% yield, colorless oil). 1 H NMR (400MHz, CDCl3) δ3.67(s,3H),3.63-3.59(m,2H),3.51-3.43(m,4H),2.52(d,1H),1.18(s,6H),0.89(s,9H),0.06(s,6H)ppm. LC-MS m / z(ESI)=321.5[M+H] + .

[0492] Step 4: Preparation of methyl 3-(2-acetoxy-3-((tert-butyldimethylsilyl)oxy)propoxy)-2,2-dimethylpropionate (compound 30e)

[0493] Under nitrogen protection, methyl 3-(3-((tert-butyldimethylsilyl)oxy)-2-hydroxypropoxy)-2,2-dimethylpropionate (compound 30d) (4.40 g, 13.73 mmol), dichloromethane (60 mL), diisopropylethylamine (3.55 g, 27.20 mmol), and acetyl chloride (1.29 g, 16.47 mmol) were added sequentially to a reaction flask at 0 °C. After the addition was complete, the mixture was stirred for 0.5 hours. The reaction was monitored by TLC until complete. The mixture was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain methyl 3-(2-acetoxy-3-((tert-butyldimethylsilyl)oxy)propoxy)-2,2-dimethylpropionate (compound 30e) (4.0 g, 80% yield, colorless oil). LC-MS m / z (ESI) = 363.5 [M+H] + .

[0494] Step 5: Preparation of methyl 3-(2-acetoxy-3-hydroxypropoxy)-2,2-dimethylpropionate (compound 30f)

[0495] Under nitrogen protection, methyl 3-(2-acetoxy-3-((tert-butyldimethylsilyl)oxy)propoxy)-2,2-dimethylpropionate (compound 30e) (2.40 g, 6.62 mmol), tetrahydrofuran (30 mL), and hydrochloric acid aqueous solution (2 mL, 1.0 N) were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. The mixture was extracted with ethyl acetate (100 mL × 2), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give methyl 3-(2-acetoxy-3-hydroxypropoxy)-2,2-dimethylpropionate (compound 30f) (1.2 g, 73% yield, colorless oil). LC-MS m / z (ESI) = 249.2 [M+H] + .

[0496] Step 6: Preparation of methyl 3-(2-acetoxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionate (compound 30g)

[0497] Under nitrogen protection, acetic anhydride (20 mL) and concentrated nitric acid (5 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 minutes to prepare a mixed acid solution. A solution of methyl 3-(2-acetoxy-3-hydroxypropoxy)-2,2-dimethylpropionate (compound 30f) (1.20 g, 4.83 mmol) in ethyl acetate (20 mL) was added dropwise, and stirring continued for 0.5 hours after the addition was complete. The reaction was monitored by TLC until complete. The reaction was quenched with water (100 mL), and the mixture was separated. The extract was obtained by extraction with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain methyl 3-(2-acetoxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionate (30 g) (1.1 g, yield 78%, colorless oil). LC-MS m / z (ESI) = 294.2 [M+H] + .

[0498] Step 7: Preparation of 3-(2-hydroxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid (compound 30h)

[0499] Under nitrogen protection, methyl 3-(2-acetoxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid (compound 30 g) (0.80 g, 2.73 mmol), tetrahydrofuran (10 mL), water (2 mL), and sodium hydroxide (0.16 g, 4.10 mmol) were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (50 mL × 3), the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 3-(2-hydroxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid (compound 30 h) (0.23 g, yield 36%, colorless oil). LC-MS m / z (ESI) = 236.2 [MH] - .

[0500] Step 8: Preparation of 3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid (compound 30i)

[0501] Under nitrogen protection, 3-(2-hydroxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid (compound 30h) (0.23 g, 0.97 mmol), dichloromethane (10 mL), imidazole (0.20 g, 2.91 mmol), and tert-butyldimethylchlorosilane (0.37 g, 2.43 mmol) were added sequentially to the reaction flask at 0 °C. After the addition was complete, the mixture was returned to room temperature and stirred for 0.5 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and methanol (10 mL) and potassium carbonate (0.15 g, 1 mL) were added. 0.10 mmol), stirred at room temperature for 1 hour; concentrated under reduced pressure, pH adjusted to 4 with hydrochloric acid aqueous solution (1N), extracted with ethyl acetate (30 mL × 3), the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid (compound 30i) (0.27 g, yield 79%, colorless oil). LC-MS m / z (ESI) = 350.3 [MH] - .

[0502] Step 9: Preparation of methyl chloro 3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionate (compound 30j)

[0503] Under nitrogen protection, at 0°C, 3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid (compound 30i) (0.27 g, 0.77 mmol), dichloromethane (10 mL), water (10 mL), sodium bicarbonate (0.32 g, 3.84 mmol), and tetrabutylammonium bisulfate (0.05 g, 0.15 mmol) were added sequentially to the reaction flask, followed by dropwise addition of chloromethylsulfonyl chloride (0.16 g). 1.00 mmol), after the addition was complete, the mixture was returned to room temperature and stirred for 1 hour; the reaction was monitored by TLC until complete; the mixture was separated, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by medium pressure to obtain compound 3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionic acid chloromethyl ester (compound 30j) (0.16 g, yield 52%, colorless oil).

[0504] Step 10: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionate (compound 30k)

[0505] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (0.06 g, 0.15 mmol), methyltetrahydrofuran (2 mL), methyl 3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionate chloromethyl ester (compound 30j) (0.08 g, 0.19 mmol) and cesium carbonate (0.06 g, 0.18 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionate (compound 30k) (0.082 g, yield 72%, white solid). LC-MS m / z (ESI) = 771.9 [M+H] + .

[0506] Step 11: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2-hydroxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionate hydrochloride (compound 30)

[0507] p-Toluenesulfonic acid hydrate (0.077 g, 0.404 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-(2-((tert-butyldimethylsilyl)oxy)-3-(nitrooxy)propoxy)-2,2-dimethylpropionate (compound 30k) (0.078 g, 0.101 mmol) in acetonitrile (5 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (1.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2-hydroxy-3-(nitrooxy)propoxy)-2,2-dimethylpropionate hydrochloride (compound 30) (41 mg, yield 68%, white solid). 1H NMR(400MHz,DMSO-d6)δ8.71(dd,1H),8.52(s,3H),8.31(dd,1H),7.50(t,1H),7.16(d,1H),5.90(d,2H),5.28-5.19(m,2H),4.60(m,1H),4. 41(dd,1H),3.90-3.84(m,1H),3.65-3.52(m,5H),3.41-3.32(m,2H),3.24(dd,1H),2.68(s,3H),2.20(m,1H),1.85(m,1H),1.18(d,6H)ppm. LC-MS m / z(ESI)=557.4[M+H] + .

[0508] [Amended according to Rule 26, 31.07.2025] Example 31: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-4-(nitrooxy)butyrate hydrochloride (compound 31)

[0509] Step 1: Preparation of sodium 4-hydroxy-2,2-dimethylbutyrate (compound 31b)

[0510] Under nitrogen protection, 3,3-dimethyldihydrofuran-2(3H)-one (compound 31a) (10.00 g, 87.61 mmol), ethanol (150 mL), water (60 mL), and sodium hydroxide (3.33 g, 83.23 mmol) were added sequentially to the reaction flask, and the mixture was stirred at 80 °C for 5 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure to give sodium 4-hydroxy-2,2-dimethylbutyrate (compound 31b) (13.5 g, 99% yield, white solid). LC-MS m / z (ESI) = 131.1 [MH] - .

[0511] Step 2: Preparation of 2,2-dimethyl-4-(nitoxy)butyric acid (compound 31c)

[0512] Under nitrogen protection, acetic anhydride (20 mL) and nitric acid (5 mL) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 min to prepare a mixed acid solution. A solution of sodium 4-hydroxy-2,2-dimethylbutyrate (compound 31b) (1.00 g, 6.49 mmol) in ethyl acetate (20 mL) was added dropwise, and stirring continued for 1 hour after the addition was complete. The reaction was monitored by TLC until complete. The reaction was quenched with water (50 mL), extracted with ethyl acetate (50 mL × 2), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 2,2-dimethyl-4-(nitoxy)butyric acid (compound 31c) (0.13 g, yield 11%, colorless oil). LC-MS m / z (ESI) = 176.1 [MH] - .

[0513] Step 3: Preparation of chloromethyl 2,2-dimethyl-4-(nitoxy)butyrate (compound 31d)

[0514] Under nitrogen protection, 2,2-dimethyl-4-(nitoxy)butyric acid (compound 31c) (0.13 g, 0.73 mmol), dichloromethane (5 mL), water (5 mL), sodium bicarbonate (0.31 g, 3.67 mmol), and tetrabutylammonium bisulfate (0.05 g, 0.15 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (0.16 g, 0.95 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 2,2-dimethyl-4-(nitoxy)butyrate (compound 31d) (0.11 g, yield 85%, colorless oil).

[0515] Step 4: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-4-(nitrooxy)butyrate (compound 31e)

[0516] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.10 g, 0.25 mmol), methyltetrahydrofuran (5 mL), methyl 2,2-dimethyl-4-(nitoxy)butyrate (compound 31d) (0.08 g, 0.37 mmol) and cesium carbonate (0.10 g, 0.29 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-4-(nitrooxy)butyrate (compound 31e) (0.13 g, yield 89%, white solid). LC-MS m / z (ESI) = 597.5 [M+H] + .

[0517] Step 5: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-4-(nitrooxy)butyrate hydrochloride (compound 31)

[0518] p-Toluenesulfonic acid hydrate (0.166 g, 0.870 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-4-(nitrooxy)butyrate (compound 31e) (0.13 g, 0.22 mmol) in acetonitrile (5 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-4-(nitrooxy)butyrate hydrochloride (compound 31) (71 mg, yield 61%, white solid). 1H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.11(m,4H),7.69(t,1H),7.55(s,1H),5.90(s,2H),4.53(t,2H),4.04(m,1H) ,3.76-3.67(m,2H),3.54(m,1H),3.45(m,1H),2.59(s,3H),2.45(m,1H),2.13(m,1H),1.92(t,2H),1.17(s,6H)ppm. LC-MS m / z(ESI)=497.2[M+H] + .

[0519] [Amended according to Rule 26, 31.07.2025] Example 32: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-6-(nitrooxy)hexanoate hydrochloride (compound 32)

[0520] Step 1: Preparation of ethyl 2,2-dimethyl-6-(nitoxy)hexanoate (compound 32b)

[0521] Under nitrogen protection, ethyl 6-bromo-2,2-dimethylhexanoate (compound 32a) (3.00 g, 11.95 mmol), silver nitrate (4.06 g, 23.89 mmol), and acetonitrile (30 mL) solution were added sequentially to the reaction flask. The mixture was stirred at 75 °C for 16 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, and the residue was separated by medium-pressure preparative chromatography to give compound 2,2-dimethyl-6-(nitoxy)hexanoate (compound 32b) (2.6 g, 93% yield, colorless oil). LC-MS m / z (ESI) = 234.2 [M+H] + .

[0522] Step 2: Preparation of 2,2-dimethyl-6-(nitrooxy)hexanoic acid (compound 32c)

[0523] Under nitrogen protection, ethyl 2,2-dimethyl-6-(nitoxy)hexanoate (compound 32b) (2.60 g, 11.15 mmol), tetrahydrofuran (30 mL), water (5 mL), and sodium hydroxide (0.67 g, 16.72 mmol) were added sequentially to the reaction flask. The mixture was stirred at 60 °C for 8 hours. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 2,2-dimethyl-6-(nitoxy)hexanoic acid (compound 32c) (1.5 g, 66% yield, colorless oil). LC-MS m / z (ESI) = 204.2 [MH] - .

[0524] Step 3: Preparation of chloromethyl 2,2-dimethyl-6-(nitoxy)hexanoate (compound 32d)

[0525] Under nitrogen protection, 2,2-dimethyl-6-(nitoxy)hexanoic acid (compound 32c) (1.50 g, 7.31 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (3.07 g, 36.55 mmol), and tetrabutylammonium bisulfate (0.50 g, 1.46 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl sulfonyl chloride (1.57 g, 9.50 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 2,2-dimethyl-6-(nitoxy)hexanoate (compound 32d) (1.1 g, yield 59%, colorless oil). 1 H NMR (500MHz, CDCl3) δ5.72(s,2H),4.44(t,2H),1.75-1.68(m,2H),1.62-1.55(m,2H),1.40-1.32(m,2H),1.21(s,6H)ppm.

[0526] Step 4: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-6-(nitrooxy)hexanoate (compound 32e)

[0527] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), methyl 2,2-dimethyl-6-(nitrooxy)hexanoate (compound 32d) (0.37 g, 1.47 mmol) and cesium carbonate (0.48 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2,2-dimethyl-6-(nitrooxy)hexanoate (compound 32e) (0.68 g, yield 88%, white solid). LC-MS m / z (ESI) = 625.6 [M+H] + .

[0528] Step 5: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-6-(nitrooxy)hexanoate hydrochloride (compound 32)

[0529] p-Toluenesulfonic acid hydrate (0.81 g, 4.28 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-6-(nitrooxy)hexanoate (compound 32e) (0.67 g, 1.07 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2,2-dimethyl-6-(nitrooxy)hexanoate hydrochloride (compound 32) (510 mg, yield 85%, white solid). 1H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.54(s,3H),8.17(d,1H),7.68(t,1H),7.55(s,1H),5.90(s,2H),4.33(t,2H),4.09-4.00(m,1H) ,3.80-3.66(m,2H),3.59-3.45(m,2H),2.59(s,3H),2.45(m,1H),2.19(m,1H),1.55-1.44(m,4H),1.21-1.13(m,2H),1.11(s,6H)ppm. LC-MS m / z(ESI)=525.3[M+H] + .

[0530] [Amended according to Rule 26, 31.07.2025] Example 33: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]1-pentane-1-carboxylate (compound 33)

[0531] Step 1: Preparation of methyl 3-hydroxymethylbicyclo[1.1.1]pentane-1-carboxylic acid (compound 33b)

[0532] 3-(methoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33a) (4.5 g, 26.4 mmol) was added to a 250 mL three-necked flask, tetrahydrofuran (50 mL) was added, and under nitrogen protection, boranetetrahydrofuran solution (31.7 mL, 31.7 mmol, 1.0 N) was added and stirred for 3 hours. The reaction was monitored by TLC until complete, quenched with water, concentrated under reduced pressure, extracted with ethyl acetate (100 mL), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. After preparative separation under medium pressure, compound 3-hydroxymethylbicyclo[1.1.1]pentane-1-carboxylic acid methyl ester (compound 33b) (3.5 g, yield 85%, colorless oil) was obtained. 1 H NMR (500MHz, CDCl3) δ3.70 (s, 3H), 3.65 (s, 2H), 2.02 (s, 6H) ppm.

[0533] Step 2: Preparation of methyl 3-formylbicyclo[1.1.1]pentane-1-carboxylate (compound 33c)

[0534] Under nitrogen protection, methyl 3-hydroxymethylbicyclo[1.1.1]pentane-1-carboxylate (compound 33b) (1.00 g, 6.40 mmol), dichloromethane (20 mL), and Desmartin oxidant (2.72 g, 6.40 mmol) were added sequentially to the reaction flask at 0 °C. After the addition was complete, the mixture was returned to room temperature and stirred for 0.5 hours. The reaction was monitored by TLC until complete. The organic phase was washed with saturated sodium bicarbonate solution (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparative separation to obtain methyl 3-formylbicyclo[1.1.1]pentane-1-carboxylate (compound 33c) (0.75 g, 76% yield, colorless oil).

[0535] Step 3: Preparation of methyl 3-(1-hydroxyethyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 33d)

[0536] Under nitrogen protection, methyl 3-formylbicyclo[1.1.1]pentane-1-carboxylate (compound 33c) (0.75 g, 4.87 mmol), tetrahydrofuran (10 mL), and methyl magnesium bromide (4.87 mL, 4.87 mmol, 1.0 N) were added sequentially to the reaction flask at 0 °C. After addition, stirring was continued for ten minutes. The reaction was monitored by TLC until complete. The organic phase was washed with saturated ammonium chloride solution (10 mL), extracted with ethyl acetate (100 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound methyl 3-(1-hydroxyethyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 33d) (0.41 g, 50% yield, colorless oil). LC-MS m / z (ESI) = 171.2 [M+H] + .

[0537] Step 4: Preparation of 3-(1-hydroxyethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33e)

[0538] Under nitrogen protection, methyl 3-(1-hydroxyethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33d) (2.00 g, 11.75 mmol), tetrahydrofuran (30 mL), water (5 mL), and lithium hydroxide (0.56 g, 23.50 mmol) were added sequentially to the reaction flask, and the mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC until complete. The pH was adjusted to 2 with 1 N hydrochloric acid, the mixture was concentrated under reduced pressure, extracted with ethyl acetate (100 mL × 2), the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 3-(1-hydroxyethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33e) (1.4 g, 76% yield, colorless oil). LC-MS m / z (ESI) = 155.1 [MH] - .

[0539] Step 5: Preparation of 3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33f)

[0540] Under nitrogen protection, dichloromethane (50 mL), concentrated nitric acid (17 mL), and concentrated sulfuric acid (17 mL) were added sequentially to the reaction flask at 0 °C. The mixture was stirred for 30 min to prepare a mixed acid solution. A solution of 3-(1-hydroxyethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33e) (1.20 g, 7.68 mmol) in dichloromethane (20 mL) was added dropwise. After the addition was complete, the mixture was stirred for 1 hour. The reaction was monitored by TLC until it was complete. The reaction was quenched with water (50 mL), and the mixture was extracted with dichloromethane (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain compound 3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33f) (1.2 g, yield 78%, colorless oil). LC-MS m / z (ESI) = 200.1 [MH] - .

[0541] Step 6: Preparation of methyl chloromethyl 3-(1-(nitoxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 33g)

[0542] Under nitrogen protection, 3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33f) (1.20 g, 5.97 mmol), dichloromethane (40 mL), water (40 mL), sodium bicarbonate (2.5 g, 29.8 mmol), and tetrabutylammonium bisulfate (0.41 g, 1.19 mmol) were added dropwise to the reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain chloromethyl 3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 33 g) (0.7 g, yield 47%, colorless oil). 1 H NMR (400MHz, CDCl3) δ5.73(s,2H),5.15(m,1H),2.19-2.09(m,6H),1.33(d,3H)ppm.

[0543] Step 7: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 33h)

[0544] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), methyl 3-(1-(nitoxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 33 g) (0.46 g, 1.84 mmol) and cesium carbonate (0.47 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 33h) (0.49 g, yield 64%, white solid). LC-MS m / z (ESI) = 621.4 [M+H] + .

[0545] Step 8: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]1-pentane-1-carboxylate (compound 33)

[0546] p-Toluenesulfonic acid hydrate (0.60 g, 3.16 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 33h) (0.49 g, 0.79 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure. The product was added to ethyl acetate hydrochloride solution (2.0 mL, 4 N) and concentrated under reduced pressure to give (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(1-(nitrooxy)ethyl)bicyclo[1.1.1]1-pentane-1-carboxylate (compound 33) (195 mg, yield 44%, white solid). 1 H NMR(400MHz,DMSO-d6)δ8.65(d,1H),8.35(s,3H),8.13(d,1H),7.71(t,1H),7.56(s,1H),5.91(s,2H),5.20(m,1H),4.05 (m,1H),3.72(m,2H),3.58-3.44(m,2H),2.59(s,3H),2.49-2.42(m,1H),2.14(m,1H),2.04-1.96(m,6H),1.23(d,3H)ppm. LC-MS m / z(ESI)=521.3[M+H] + .

[0547] [Amended according to Rule 26, 31.07.2025] Example 34: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate hydrochloride (compound 34)

[0548] Step 1: Preparation of methyl 3-((allyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 34a)

[0549] Under nitrogen protection, methyl 3-hydroxymethylbicyclo[1.1.1]pentane-1-carboxylate (compound 33b) (5.50 g, 35.22 mmol), tetrahydrofuran (80 mL), and sodium hydride (1.69 g, 42.26 mmol, 60%) were added sequentially to the reaction flask at 0 °C, and the mixture was stirred for 30 minutes. Allyl bromide (6.39 g, 52.82 mmol) was added dropwise, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and saturated ammonium chloride solution (100 mL) was added dropwise to quench the reaction. The aqueous phase was extracted with ethyl acetate (100 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated under medium pressure to obtain methyl 3-((allyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 34a) (2.3 g, yield 33%, colorless oil). LC-MS m / z (ESI) = 197.1 [M+H] + .

[0550] Step 2: Preparation of 3-((3-hydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 34b)

[0551] At 0°C, under nitrogen protection, a solution of borane tetrahydrofuran (15.3 mL, 15.3 mmol, 1N) was added dropwise to a solution of methyl 3-((allyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 34a) (2.00 g, 10.19 mmol) in tetrahydrofuran (20 mL). The resulting mixture was heated to room temperature and stirred for 1 hour. The mixture was cooled to 0°C, and sodium hydroxide aqueous solution (1.43 g, 35.67 mmol, 2N) and hydrogen peroxide (3.47 g, 30.57 mmol, 30%) were added dropwise. After the addition was complete, the mixture was kept at this temperature for 30 min, then returned to room temperature and stirred for 1 h. The mixture was then cooled to 0°C, and the reaction was quenched by adding saturated sodium bisulfite solution (100 mL). The pH was adjusted to 2 with hydrochloric acid aqueous solution, and the mixture was extracted with ethyl acetate (200 mL × 2). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 3-((3-hydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 34b) (1.8 g, yield 88%, colorless oil). LC-MS m / z (ESI) = 199.2 [MH]. - .

[0552] Step 3: Preparation of 3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 34c)

[0553] Under nitrogen protection, dichloromethane (50 mL), concentrated nitric acid (17 mL), and concentrated sulfuric acid (17 mL) were added sequentially to the reaction flask at 0 °C. The mixture was stirred for 30 min to prepare a mixed acid solution. A solution of 3-((3-hydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (34b) (1.80 g, 8.99 mmol) in dichloromethane (20 mL) was added dropwise. After the addition was complete, the mixture was stirred for 1 hour. The reaction was monitored by TLC until it was complete. The reaction was quenched with water (50 mL), and the mixture was extracted with dichloromethane (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain compound 3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 34c) (1.6 g, yield 73%, colorless oil). LC-MS m / z (ESI) = 244.2 [MH] - .

[0554] Step 4: Preparation of chloromethyl 3-((3-(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 34d)

[0555] Under nitrogen protection, at 0°C, 3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 34c) (1.60 g, 6.52 mmol), dichloromethane (40 mL), water (40 mL), sodium bicarbonate (2.74 g, 32.62 mmol), and tetrabutylammonium bisulfate (0.44 g, 1.31 mmol) were added sequentially to the reaction flask, followed by dropwise addition of chloromethylsulfonyl chloride (1.6 g, 6.52 mmol). 1 g (9.79 mmol), after which the mixture was added, the mixture was brought back to room temperature and stirred for 2 hours; the reaction was monitored by TLC until complete; the mixture was separated, the organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by medium pressure to obtain compound 3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 34d) (1.6 g, yield 84%, colorless oil).

[0556] Step 5: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester (compound 34e)

[0557] Under nitrogen protection, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.50 g, 1.23 mmol), methyltetrahydrofuran (10 mL), 3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 34d) (0.54 g, 1.84 mmol) and cesium carbonate (0.47 g, 1.47 mmol) were added sequentially to the reaction flask, and the mixture was heated to 55 °C and stirred for 2 hours. The reaction was monitored by TLC until complete; the mixture was filtered, dried over anhydrous sodium sulfate, filtered again, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester (compound 34e) (0.71 g, yield 87%, white solid). LC-MS m / z (ESI) = 665.6 [M+H] + .

[0558] Step 6: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate hydrochloride (compound 34)

[0559] p-Toluenesulfonic acid hydrate (0.81 g, 4.27 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester (compound 34e) (0.71 g, 1.07 mmol) in acetonitrile (10 mL) at room temperature and stirred for 0.5 hours at room temperature. The solution was concentrated under reduced pressure, and the pH was adjusted to 8 by adding sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (50 mL). The combined organic phases were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was then separated by high pressure. The product was added to ethyl acetate hydrochloride solution (1.0 mL, 4 N) and concentrated under reduced pressure to obtain (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-((3-(nitrooxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate hydrochloride (compound 34) (120 mg, yield 19%, white solid). 1H NMR(400MHz,DMSO-d6)δ8.76(d,1H),8.35(s,3H),8.31(d,1H),7.51(t,1H),7.19(s,1H),5.92(s,2H),4.54(t,2H),3.89-3.82(m,1H),3. 73-3.60(m,3H),3.50(t,2H),3.40(s,2H),3.24(dd,1H),2.71(s,3H),2.23(m,1H),2.01(s,6H),1.99-1.93(m,2H),1.90-1.82(m,1H)ppm. LC-MS m / z(ESI)=565.4[M+H] + .

[0560] [Amended according to Rule 26, 31.07.2025] Example 35: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-((nitrooxy)methyl)cyclohexane-1-carboxylic acid hydrochloride (compound 35)

[0561] Step 1: Preparation of (1S,4S)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid (compound 35b)

[0562] At 0°C, nitric acid (20 mL) was slowly added to a sulfuric acid (20 mL) solution and stirred for 50 minutes. A solution of (1S,4S)-4-(hydroxymethyl)cyclohexane-1-carboxylic acid (compound 35a) (2.5 g, 15.8 mmol) in dichloromethane (20 mL) was added dropwise. The resulting mixture was stirred at 0°C for 1.5 hours. The reaction was monitored by TLC until complete. The reaction solution was slowly poured into ice water (300 mL) to quench the reaction. The aqueous phase was extracted with dichloromethane (100 mL × 2), washed with saturated brine (50 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparative separation to obtain compound (1S,4S)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid (compound 35b) (3.0 g, 94% yield, white solid). 1 H NMR (400MHz, CDCl3) δ4.35(d,2H),2.76-2.66(m,1H),2.17-2.07(m,2H),1.98-1.84(m,1H),1.78-1.59(m,4H),1.48-1.35(m,2H)ppm.

[0563] Step 2: Preparation of (1S,4S)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid chloromethyl ester (compound 35c)

[0564] Under nitrogen protection, (1S,4S)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid (compound 35b) (1.50 g, 7.38 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (3.1 g, 36.9 mmol), and tetrabutylammonium bisulfate (0.50 g, 1.48 mmol) were added dropwise to a reaction flask at 0 °C. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated into liquid and liquid phases. The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (1S,4S)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid chloromethyl ester (compound 35c) (1.3 g, 70% yield, white solid). 1 H NMR (400MHz, CDCl3) δ5.77(d,2H),4.34(dd,2H),2.80-2.67(m,1H),2.19-2.05(m,2H),1.98-1.83(m,1H),1.77-1.59(m,4H),1.45-1.27(m,2H)ppm.

[0565] Step 3: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-((nitro)methyl)cyclohexane-1-carboxylic acid ester (compound 35d)

[0566] Under nitrogen protection, at room temperature, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (0.35 g, 0.86 mmol), 2-methyltetrahydrofuran (20 mL), cesium carbonate (0.42 g, 1.29 mmol), and (1S,4S)-4-((nitoxy)methyl)cyclohexane-1-carboxylic acid chloromethyl ester (compound 35c) (0.43 g, 1.72 mmol) were added sequentially to a reaction flask. The resulting mixture was heated to 55 °C and stirred for 1 hour. The reaction was monitored by TLC until complete; the mixture was cooled to room temperature, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-((nitrooxy)methyl)cyclohexane-1-carboxylic acid ester (compound 35d) (0.5 g, 93% yield, white solid). LC-MS m / z (ESI) = 623.1 [M+H] + .

[0567] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-((nitrooxy)methyl)cyclohexane-1-carboxylic acid hydrochloride (compound 35)

[0568] p-Toluenesulfonic acid hydrate (0.61 g, 3.20 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-((nitrooxy)methyl)cyclohexane-1-carboxylic acid ester (compound 35d) (0.50 g, 0.80 mmol) in acetonitrile (10 mL) at room temperature. The resulting mixture was stirred at room temperature for 30 minutes. Concentrate under reduced pressure, adjust to alkalinity by adding saturated sodium bicarbonate aqueous solution (10 mL), extract the aqueous phase with ethyl acetate (30 mL × 2), wash with saturated brine (20 mL × 2), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate the residue under medium pressure to obtain compound (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-((nitrooxy)methyl)cyclohexane-1-carboxylic acid ester; at room temperature, add 0.5 mL of ethyl acetate hydrochloride solution (4N) to (5-( The compound (5-((((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,4R)-4-((nitrooxy)methyl)cyclohexane-1-carboxylic acid ester) was concentrated under reduced pressure in a solution of dichloromethane (10 mL) to give compound (Compound 35) (0.36 g, 80% yield, white solid). 1H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.61-8.38(m,3H),8.18(s,1H),7.69( t,1H),7.56(s,1H),5.92(s,2H),4.35(d,2H),4.10-3.97(m,1H),3.81-3.64 (m,2H),3.61-3.44(m,2H),2.67(m,1H),2.59(s,3H),2.48-2.40(m,1H),2.2 6-2.10(m,1H),1.96-1.75(m,3H),1.62-1.46(m,4H),1.29-1.19(m,2H)ppm. LC-MS m / z (ESI) = 523.1 [M+H] + .

[0569] [Amended according to Rule 26, 31.07.2025] Example 36: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-(1-((nitrooxy)methyl)cyclopropyl)acetic acid hydrochloride (compound 36)

[0570] Step 1: Preparation of 2-(1-(hydroxymethyl)cyclopropyl)acetic acid (compound 36b)

[0571] 2-(1-(hydroxymethyl)cyclopropyl)acetonitrile (compound 36a) (5.0 g, 45.0 mmol) was added to a mixture of ethanol (50 mL) and water (6 mL), followed by the addition of potassium hydroxide (7.6 g, 135.0 mmol). The mixture was stirred at 80 °C for 6 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure, cooled to 0 °C, and the pH was adjusted to 1–2 with dilute hydrochloric acid. The aqueous phase was extracted with ethyl acetate (50 mL × 2), washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 2-(1-(hydroxymethyl)cyclopropyl)acetic acid (compound 36b) (2.5 g, 43% yield). 1 H NMR (500MHz, DMSO-d6) δ3.28(s,2H),2.27(s,2H),0.44-0.31(m,4H)ppm.

[0572] Step 2: Preparation of 2-(1-((nitoxy)methyl)cyclopropyl)acetic acid (compound 36c)

[0573] Nitric acid (5 mL) was slowly added to a 10 mL solution of acetic anhydride at 0 °C and stirred for 30 minutes. This solution was then slowly added dropwise to a 60 mL solution of acetic anhydride containing 1.0 g (7.7 mmol) of 2-(1-(hydroxymethyl)cyclopropyl)acetic acid (compound 36b). The resulting mixture was stirred at 0 °C for 1 hour. The reaction was monitored by TLC until complete. The reaction solution was quenched by slowly pouring it into 200 mL of ice water. The aqueous phase was extracted with dichloromethane (50 mL × 2), washed with saturated brine (30 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound 2-(1-((nitoxy)methyl)cyclopropyl)acetic acid (compound 36c) (1.3 g, 96% yield, colorless liquid). LC-MS m / z (ESI) = 174.1 [MH] - .

[0574] Step 3: Preparation of 2-(1-((nitoxy)methyl)cyclopropyl)chloromethyl acetate (compound 36d)

[0575] Under nitrogen protection, 2-(1-((nitoxy)methyl)cyclopropyl)acetic acid (compound 36c) (1.3 g, 7.4 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (3.1 g, 37.0 mmol), and tetrabutylammonium bisulfate (0.5 g, 1.5 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl chlorosulfonate (1.5 g, 8.9 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain chloromethyl 2-(1-((nitoxy)methyl)cyclopropyl)acetic acid (compound 36d) (0.57 g, yield 35%, colorless oil). 1 H NMR (500MHz, CDCl3) δ5.74(s,2H),4.44(s,2H),2.48(s,2H),0.83-0.68(m,4H)ppm.

[0576] Step 4: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-(1-((nitrooxy)methyl)cyclopropyl)acetate (compound 36e)

[0577] Under nitrogen protection, at room temperature, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.2 g, 0.5 mmol), 2-methyltetrahydrofuran (10 mL), cesium carbonate (0.24 g, 0.75 mmol), and 2-(1-((nitoxy)methyl)cyclopropyl)chloromethyl acetate (compound 36d) (0.34 g, 1.50 mmol) were added sequentially to a reaction flask. The resulting mixture was heated to 55 °C and stirred for 1 hour. The reaction was monitored by TLC until complete; the mixture was cooled to room temperature, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-(1-((nitrooxy)methyl)cyclopropyl)acetate (compound 36e) (0.24 g, yield 81%, white solid). LC-MS m / z (ESI) = 595.2 [M+H] + .

[0578] Step 5: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-(1-((nitrooxy)methyl)cyclopropyl)acetic acid hydrochloride (compound 36)

[0579] p-Toluenesulfonic acid hydrate (0.61 g, 3.23 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-(1-((nitrooxy)methyl)cyclopropyl)acetate (compound 36e) (0.24 g, 0.40 mmol) in acetonitrile (5 mL) at room temperature. The resulting mixture was stirred at room temperature for 30 minutes. The solution was concentrated under reduced pressure, and the solution was adjusted to alkalinity by adding 5 mL of saturated sodium bicarbonate solution. The aqueous phase was extracted with 20 mL of ethyl acetate (22 mL × 2), washed with 10 mL of saturated brine (22 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-2-(1-((nitro)methyl)cyclopropyl)acetate. The ethyl acetate hydrochloride solution (0.5 mL, 4N) was added dropwise at room temperature. The solution of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2-(1-((nitrooxy)methyl)cyclopropyl)acetate was concentrated under reduced pressure in dichloromethane (10 mL) to give (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl 2-(1-((nitrooxy)methyl)cyclopropyl)acetate hydrochloride (compound 36) (0.11 g, 52% yield, white solid). 1 H NMR(400MHz,DMSO-d6)δ8.65(d,1H),8.49(s,3H),8.18(d,1H),7.71(t,1H),7.55(s,1H),5.92(s,2H),4.47(s,2H),4 .06(s,1H),3.85-3.63(m,2H),3.60-3.45(m,2H),2.60(s,3H),2.49-2.41(m,3H),2.25-2.13(m,1H),0.66(m,4H)ppm. LC-MS m / z(ESI)=495.1[M+H] + .

[0580] [Amended according to Rule 26, 31.07.2025] Example 37: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid hydrochloride (compound 37)

[0581] Step 1: Preparation of methyl 3-((allyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37b)

[0582] Under nitrogen protection, sodium hydride (1.5 g, 38.4 mmol, 60%) was added to a tetrahydrofuran (50 mL) solution of methyl 3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37a) (5.0 g, 32.0 mmol) at 0 °C. The mixture was stirred for 30 minutes, and then 3-bromoprop-1-ene (5.8 g, 48.0 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 4 hours. The reaction was monitored by TLC until complete. The temperature was lowered to 0 °C, and the reaction was quenched by adding water (20 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 2), washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give methyl 3-((allyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37b) (2.2 g, yield 35%, white solid). 1 H NMR (500MHz, CDCl3) δ5.92-5.83(m,1H),5.28-5.14(m,2H),3.97(d,2H),3.66(s,3H),3.43(s,2H),2.01(s,6H)ppm.

[0583] Step 2: Preparation of methyl 3-((2,3-dihydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37c)

[0584] At 0°C, AD-mixture-ALPHA (15.0 g) was added to a mixed solution of tert-butanol (60 mL) and water (60 mL), followed by methyl 3-((allyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 37b) (2.20 g, 11.21 mmol). The resulting mixture was stirred at 0°C for 6 hours. The reaction was monitored by TLC until complete. The temperature was lowered to 0°C, and the reaction was quenched by adding saturated sodium sulfite (20 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 2), washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain methyl 3-((2,3-dihydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (compound 37c) (1.7 g, yield 66%, white solid). 1H NMR (400MHz, CDCl3) δ3.88(s,1H),3.70(s,3H),3.56(t,2H),3.52(s,2H),2.77(d,1H),2.39(s,1H),2.04(s,6H)ppm. LC-MS m / z(ESI)=231.1[M+H] + .

[0585] Step 3: Preparation of methyl 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37d)

[0586] Nitric acid (15 mL) was slowly added to concentrated sulfuric acid (8 mL) solution at 0 °C and stirred for 30 minutes. A solution of methyl 3-((2,3-dihydroxypropoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37c) (1.6 g, 6.9 mmol) in dichloromethane (10 mL) was added dropwise. The resulting mixture was stirred at 0 °C for 1 hour. The reaction was monitored by TLC until complete. The reaction solution was slowly poured into ice water (200 mL) to quench the reaction. The aqueous phase was extracted with ethyl acetate (50 mL × 2), washed with saturated brine (30 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium pressure to obtain methyl 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37d) (2.0 g, 90% yield, colorless liquid). 1 H NMR (500MHz, CDCl3) δ5.41(s,1H),4.82(d,1H),4.69-4.61(m,1H),3.78-3.71(m,2H),3.69(s,3H),3.56-3.47(m,2H),2.02(s,6H)ppm.

[0587] Step 4: Preparation of 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37e)

[0588] Methyl 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37d) (2.0 g, 6.2 mmol) was added to a mixture of methanol (20 mL), tetrahydrofuran (20 mL), and water (10 mL). Lithium hydroxide (0.3 g, 12.5 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. The reaction was monitored by TLC until complete. The mixture was evaporated to dryness under reduced pressure, diluted with water (20 mL), and the pH was adjusted to 3-4 with dilute hydrochloric acid. The aqueous phase was extracted with ethyl acetate (30 mL × 2), washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37e) (1.5 g, 79% yield). 1 H NMR (400MHz, CDCl3) δ5.47-5.39(m,1H),4.88-4.80(m,1H),4.70-4.62(m,1H),3.80-3.70(m,2H),3.57-3.49(m,2H),2.07(s,6H)ppm.

[0589] Step 5: Preparation of 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 37f)

[0590] Under nitrogen protection, at 0°C, 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid (compound 37e) (1.5 g, 4.9 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (2.1 g, 24.5 mmol), and tetrabutylammonium bisulfate (0.34 g, 1.00 mmol) were added dropwise, followed by the addition of chloromethylchlorosulfonate (1.2 g). 7.4 mmol), after the addition was complete, the mixture was returned to room temperature and stirred for 1 hour; the reaction was monitored by TLC until complete; the mixture was separated, the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by medium pressure to obtain compound 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 37f) (1.2 g, yield 71%, colorless oil).

[0591] Step 6: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester (compound 37g)

[0592] Under nitrogen protection, at room temperature, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.5 g, 1.2 mmol), 2-methyltetrahydrofuran (20 mL), cesium carbonate (0.6 g, 1.8 mmol), and 3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid chloromethyl ester (compound 37f) (0.9 g, 2.5 mmol) were added sequentially to a reaction flask. The resulting mixture was heated to 55 °C and stirred for 1 hour. The reaction was monitored by TLC until complete; the mixture was cooled to room temperature, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester (compound 37 g) (0.46 g, yield 53%, white solid). LC-MS m / z (ESI) = 726.1 [M+H] + .

[0593] Step 7: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid hydrochloride (compound 37)

[0594] p-Toluenesulfonic acid hydrate (0.97 g, 5.10 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester (compound 37 g) (0.46 g, 0.60 mmol) in acetonitrile (10 mL) at room temperature. The resulting mixture was stirred at room temperature for 30 minutes. Concentrate under reduced pressure, add saturated sodium bicarbonate aqueous solution (10 mL) dropwise to adjust to alkalinity, extract the aqueous phase with ethyl acetate (30 mL × 2), wash with saturated brine (20 mL × 2), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate the residue under medium pressure to obtain compound (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester; add 0.5 mL of ethyl acetate hydrochloride solution (4N) dropwise to (5-(((S))-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid ester at room temperature; The compound was obtained by concentrating the ester of 3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid in a solution of dichloromethane (10 mL) under reduced pressure to give compound 37 (((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl3-((2,3-bis(nitoxy)propoxy)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid hydrochloride (compound 37) (0.2 g, 50% yield, white solid). 1 H NMR(400MHz,DMSO-d6)δ8.65(d,1H),8.52(s,3H),8.19(d,1H),7.71(t,1H),7.57(s,1H),5.91(s,2H),5.65-5.51(m,1H),5.00-4.87(m,1H) ,4.84-4.71(m,1H),4.06(s,1H),3.81-3.68(m,4H),3.59-3.42(m,4H ),2.60(s,3H),2.49-2.40(m,1H),2.25-2.15(m,1H),1.94(s,6H)ppm. LC-MS m / z(ESI)=626.1[M+H] + .

[0595] [Amended according to Rule 26, 31.07.2025] Example 38: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid hydrochloride (compound 38)

[0596] Step 1: Preparation of (1R,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid (compound 38b)

[0597] Under nitrogen protection, nitric acid (2 mL) was slowly added to acetic anhydride (8 mL) solution at 0 °C and stirred for 30 minutes. A solution of ethyl acetate (5 mL) of (1R,3R)-3-hydroxycyclobutane-1-carboxylic acid (compound 38a) (1.0 g, 8.6 mmol) was added dropwise. The resulting mixture was stirred at 0 °C for 1 hour. The reaction was monitored by TLC until complete. The reaction solution was slowly poured into ice water (100 mL) to quench the reaction. The aqueous phase was extracted with ethyl acetate (20 mL × 2), washed with saturated brine (20 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound (1R,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid (compound 38b) (1.2 g, 87% yield, colorless liquid). LC-MS m / z (ESI) = 160.1 [MH] - .

[0598] Step 2: Preparation of (1R,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid chloromethyl ester (compound 38c)

[0599] Under nitrogen protection, (1R,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid (compound 38b) (1.2 g, 8.0 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (3.4 g, 40.0 mmol), and tetrabutylammonium bisulfate (0.55 g, 1.60 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl chlorosulfonate (1.6 g, 9.7 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl ester (1R,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid (compound 38c) (0.9 g, yield 53%, colorless oil). 1H NMR (400MHz, CDCl3) δ5.78(s,2H),5.49-5.40(m,1H),3.36-3.24(m,1H),2.89-2.77(m,2H),2.63-2.50(m,2H)ppm.

[0600] Step 3: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid ester (compound 38d)

[0601] Under nitrogen protection, at room temperature, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.3 g, 0.7 mmol), 2-methyltetrahydrofuran (10 mL), cesium carbonate (0.36 g, 1.10 mmol), and (1R,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid methyl ester (compound 38c) (0.4 g, 1.9 mmol) were added sequentially to a reaction flask. The resulting mixture was heated to 55 °C and stirred for 1 hour. The reaction was monitored by TLC until complete; the mixture was cooled to room temperature, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid ester (compound 38d) (0.4 g, 100% yield, white solid). LC-MS m / z (ESI) = 581.6 [M+H] + .

[0602] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid hydrochloride (compound 38)

[0603] At room temperature, p-toluenesulfonic acid hydrate (0.53 g, 2.80 mmol) was added to a solution of ((5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid ester (compound 38d) (0.4 g, 0.7 mmol) in acetonitrile (10 mL). The resulting mixture was stirred at room temperature for 30 minutes. The solution was concentrated under reduced pressure, adjusted to alkalinity by dropwise addition of saturated sodium bicarbonate aqueous solution (10 mL), and the aqueous phase was extracted with ethyl acetate (30 mL × 2). The solution was washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (5-(((S)-3-aminopyrrolidine-1-yl) (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid ester; 0.5 mL of ethyl acetate hydrochloride solution was added dropwise to (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1R,3S)-3-yl)-carboxylic acid ester at room temperature. The compound (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid hydrochloride (compound 38) (0.29 g, 80% yield, white solid) was concentrated under reduced pressure in a solution of 10 mL of dichloromethane to give compound (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1R,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid hydrochloride (compound 38) (0.29 g, 80% yield). 1 H NMR(500MHz,DMSO-d6)δ8.61(d,1H),8.51(s,3H),8.12(d,1H),7.61(t,1 H),7.52(s,1H),5.91(s,2H),5.28-5.20(m,1H),4.00-3.95(m,1H),3.71 -3.64(m,2H),3.52-3.44(m,2H),3.01-2.94(m,1H),2.70-2.62(m,2H),2 .58(s,3H),2.44-2.40(m,1H),2.34-2.28(m,2H),2.20-2.13(m,1H)ppm. LC-MS m / z(ESI)=481.2[M+H] + .

[0604] [Amended according to Rule 26, 31.07.2025] Example 39: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,3R)-3-(nitrooxy)cyclobutane-1-carboxylic acid hydrochloride (compound 39)

[0605] Step 1: Preparation of (1S,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 39b)

[0606] Under nitrogen protection, nitric acid (4 mL) was slowly added to acetic anhydride (10 mL) solution at 0 °C and stirred for 30 minutes. A solution of ethyl acetate (10 mL) of (1S,3S)-3-hydroxycyclobutane-1-carboxylic acid (compound 39a) (2.0 g, 17.2 mmol) was added dropwise. The resulting mixture was stirred at 0 °C for 1 hour. The reaction was monitored by TLC until complete. The reaction solution was slowly poured into ice water (200 mL) to quench the reaction. The aqueous phase was extracted with ethyl acetate (30 mL × 2), washed with saturated brine (30 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (1S,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 39b) (2.5 g, 90% yield, colorless liquid). LC-MS m / z (ESI) = 160.1 [MH] - .

[0607] Step 2: Preparation of (1S,3S)-3-(nitoxy)cyclobutane-1-carboxylic acid chloromethyl ester (compound 39c)

[0608] Under nitrogen protection, (1S,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 39b) (2.5 g, 15.5 mmol), dichloromethane (20 mL), water (20 mL), sodium bicarbonate (6.5 g, 77.6 mmol), and tetrabutylammonium bisulfate (1.0 g, 3.1 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl chlorosulfonate (3.1 g, 18.6 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated into liquid and liquid phases. The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl ester (1S,3S)-3-(nitrooxy)cyclobutane-1-carboxylic acid (compound 39c) (1.7 g, yield 52%, colorless oil). 1H NMR (500MHz, CDCl3) δ 5.73 (s, 2H), 5.23-5.16 (m, 1H), 2.98-2.90 (m, 1H), 2.83-2.75 (m, 2H), 2.57-2.49 (m, 2H) ppm.

[0609] Step 3: Preparation of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1S,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid ester (compound 39d)

[0610] Under nitrogen protection, at room temperature, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl) tert-butyl carbamate (compound M1) (0.3 g, 0.7 mmol), 2-methyltetrahydrofuran (10 mL), cesium carbonate (0.36 g, 1.1 mmol), and (1S,3S)-3-(nitoxy)cyclobutane-1-carboxylic acid chloromethyl ester (compound 39c) (0.7 g, 3.5 mmol) were added sequentially to a reaction flask. The resulting mixture was heated to 55 °C and stirred for 1 hour. The reaction was monitored by TLC until complete; the mixture was cooled to room temperature, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1S,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid ester (compound 39d) (0.49 g, 100% yield, white solid). LC-MS m / z (ESI) = 581.6 [M+H] + .

[0611] Step 4: Preparation of (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,3R)-3-(nitrooxy)cyclobutane-1-carboxylic acid hydrochloride (compound 39)

[0612] p-Toluenesulfonic acid hydrate (1.28 g, 6.72 mmol) was added to a solution of (5-(((S)-3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1S,3R)-3-(nitoxy)cyclobutane-1-carboxylic acid ester (compound 39d) (0.49 g, 0.80 mmol)) in 10 mL of acetonitrile at room temperature. The resulting mixture was stirred at room temperature for 30 minutes. Concentrate under reduced pressure, adjust to alkalinity by adding saturated sodium bicarbonate aqueous solution (10 mL), extract the aqueous phase with ethyl acetate (30 mL × 2), wash with saturated brine (20 mL × 2), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate the residue by medium pressure to obtain compound (5-(((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1S,3R)-3-(nitrooxy)cyclobutane-1-carboxylic acid ester; at room temperature, add 0.5 mL of ethyl acetate hydrochloride solution (4N) to (5 The compound (5-((((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)-methyl(1S,3R)-3-(nitrooxy)cyclobutane-1-carboxylic acid ester was concentrated under reduced pressure in a 10 mL solution of dichloromethane to give compound (5-((((S)-3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl(1S,3R)-3-(nitrooxy)cyclobutane-1-carboxylic acid hydrochloride (compound 39) (0.35 g, 85% yield, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.53(s,3H),8.18(d,1H),7.69(t,1 H),7.56(s,1H),5.91(s,2H),5.31-5.20(m,1H),4.08-4.00(m,1H),3.79 -3.66(m,2H),3.57-3.46(m,2H),3.04-2.95(m,1H),2.71-2.63(m,2H),2 .58(s,3H),2.48-2.40(m,1H),2.38-2.30(m,2H),2.23-2.14(m,1H)ppm. LC-MS m / z(ESI)=481.2[M+H] + .

[0613] [Amended according to Rule 26, 31.07.2025] Example 40: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitrooxy)valerate methyl ester hydrochloride (compound 40)

[0614] Step 1: Preparation of 5-bromo-2,2-dimethylvaleric acid (compound 40b)

[0615] Under nitrogen protection, at 0°C, hydrobromic acid (acetic acid solution) (7.6 g, 31.2 mmol, 33%) was added to a hexane (60 mL) solution of 2,2-dimethylpentyl-4-enoic acid (compound 40a) (2.0 g, 15.6 mmol). The resulting mixture was heated to room temperature and stirred for 4 hours. The mixture was diluted with water (50 mL), extracted with ethyl acetate (50 mL × 2), washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound 5-bromo-2,2-dimethylpentyl acid (compound 40b) (3.1 g, 95% yield, colorless liquid). 1 H NMR (400MHz, CDCl3) δ3.43(t,2H),1.97-1.85(m,2H),1.77-1.67(m,2H),1.26(s,6H)ppm.

[0616] Step 2: Preparation of 2,2-dimethyl-5-(nitrooxy)valerate (compound 40c)

[0617] Under light-protected conditions and nitrogen protection, silver nitrate (5.0 g, 29.6 mmol) was added to a solution of 5-bromo-2,2-dimethylpentanoic acid (compound 40b) (3.1 g, 14.8 mmol) in acetonitrile (50 mL). The resulting mixture was heated to reflux and stirred for 16 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (50 mL). The mixture was filtered, and the filtrate was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to give compound 2,2-dimethyl-5-(nitrooxy)pentanoic acid (compound 40c) (0.65 g, 23% yield, yellow liquid). 1 H NMR (400MHz, CDCl3) δ4.48(t,2H),1.84-1.73(m,2H),1.73-1.62(m,2H),1.26(s,6H)ppm.

[0618] Step 3: Preparation of chloromethyl 2,2-dimethyl-5-(nitoxy)valerate (compound 40d)

[0619] Under nitrogen protection, 2,2-dimethyl-5-(nitrooxy)valerate (compound 40c) (0.65 g, 3.40 mmol), dichloromethane (10 mL), water (10 mL), sodium bicarbonate (1.4 g, 17.0 mmol), and tetrabutylammonium bisulfate (0.23 g, 0.68 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl chlorosulfonate (0.84 g, 5.10 mmol) was added dropwise. After the addition was complete, the mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by TLC until complete. The mixture was separated, and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain chloromethyl 2,2-dimethyl-5-(nitrooxy)valerate (compound 40d) (0.3 g, yield 37%, colorless liquid). 1 H NMR (400MHz, CDCl3) δ5.76 (s, 2H), 4.47 (t, 2H), 1.82-1.65 (m, 4H), 1.27 (s, 6H) ppm.

[0620] Step 4: Preparation of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitrooxy)valerate (compound 40e)

[0621] Under nitrogen protection, at room temperature, (S)-(1-((4-methyl-1-oxo-1,2-dihydroisoquinoline-5-yl)sulfonyl)pyrrolidine-3-yl)carbamate tert-butyl ester (compound M1) (0.25 g, 0.61 mmol), 2-methyltetrahydrofuran (10 mL), cesium carbonate (0.30 g, 0.92 mmol), and methyl 2,2-dimethyl-5-(nitoxy)valerate (compound 40d) (0.30 g, 1.25 mmol) were added sequentially to a reaction flask. The resulting mixture was heated to 55 °C and stirred for 1 hour. The reaction was monitored by TLC until complete; the mixture was cooled to room temperature, concentrated under reduced pressure, and the residue was subjected to preparative separation under medium pressure to give compound (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitoxy)valerate (compound 40e) (0.38 g, 100% yield, white solid). LC-MS m / z (ESI) = 611.2 [M+H] + .

[0622] Step 5: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitrooxy)valerate methyl ester hydrochloride (compound 40)

[0623] At room temperature, p-toluenesulfonic acid hydrate (0.47 g, 2.48 mmol) was added to a solution of (S)-(5-((3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitrooxy)valerate (compound 40e) (0.38 g, 0.62 mmol) in acetonitrile (10 mL). The resulting mixture was stirred at room temperature for 30 minutes. The solution was concentrated under reduced pressure, and the solution was adjusted to alkalinity by adding 10 mL of saturated sodium bicarbonate solution. The aqueous phase was extracted with 30 mL × 2 ethyl acetate, washed with 20 mL × 2 saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative separation under medium pressure to obtain compound (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitrooxy)valerate. The ethyl acetate hydrochloride solution (0.5 mL, 4N) was added dropwise at room temperature. The solution of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitrooxy)valerate was concentrated under reduced pressure in dichloromethane (10 mL) to give compound (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)2,2-dimethyl-5-(nitrooxy)valerate methyl ester hydrochloride (compound 40) (0.27 g, 80% yield, white solid). 1 H NMR(500MHz,DMSO-d6)δ8.64(d,1H),8.25(s,3H),8.12(d,1H),7.68(t,1H),7.55(s,1H),5.90(s,2H),4.39(d,2H),4.09-3.99(m,1H),3 .79-3.64(m,2H),3.58-3.43(m,3H),3.29(d,1H),2.59(s,3H),2.47-2.41(m,1H),2.18-2.10(m,1H),1.29-1.20(m,2H),1.12(s,6H)ppm. LC-MS m / z(ESI)=511.2[M+H] + .

[0624] [Amended according to Rule 26, 31.07.2025] Example 41: Preparation of (S)-(5-((3-aminopyrrolidone-1-yl)sulfonyl)-4-methyl-1-oxoisoquinoline-2(1H)-yl)methyl-3-(2-(nitrooxy)ethyl)benzoate (compound 41)

[0625] Step 1: Preparation of 3-(2-(nitoxy)ethyl)benzoic acid (compound 41b)

[0626] At 0°C, nitric acid (10 mL) was added to a dichloromethane (100) solution, followed by the slow addition of concentrated sulfuric acid (2 mL) and stirring for 1 hour. Then, 3-(2-hydroxyethyl)benzoic acid (compound 41a) (2.0 g, 12.0 mmol) was added, and the resulting mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC until complete. The reaction solution was slowly poured into ice water (300 mL) to quench the reaction. The aqueous phase was extracted with dichloromethane (100 mL × 2), washed with saturated brine (50 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by medium-pressure preparation to obtain compound 3-(2-(nitoxy)ethyl)benzoic acid (compound 41b) (2.2 g, yield 88%, white solid). 1 H NMR (400MHz, CDCl3) δ8.12-7.99(m,2H),7.59-7.45(m,2H),4.73(t,2H),3.15(t,2H)ppm.

[0627] Step 2: Preparation of chloromethyl 3-(2-(nitoxy)ethyl)benzoate (compound 41c)

[0628] Under nitrogen protection, 3-(2-(nitoxy)ethyl)benzoic acid (compound 41b) (2.2 g, 10.4 mmol), dichloromethane (30 mL), water (30 mL), sodium bicarbonate (4.4 g, 52.0 mmol), and tetrabutylammonium bisulfate (0.7 g, 2.1 mmol) were added sequentially to the reaction flask at 0 °C. Chloromethyl chlorosulfonate (2.2 g, 13.5 mmol) was added dropwise. After t...

Claims

1. A compound having the structure shown in Formula (I), or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of Formula (I): in: R 1 selected from substituents of the formula -O-NO2; R 2 and R 3 are each independently selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, amino, cyano and C 1-6 alkyl; preferably from the group consisting of H, D, F, Cl, Br, I, OH, NH2, CN and C 1-3 alkyl; R 4 selected from hydrogen, deuterium, halogen, hydroxyl, amino, cyano and optionally substituted C a alkyl; preferably selected from H, D, F, CI, Br, I, OH, NH2, CN and optionally substituted C 1-6 alkyl; preferably selected from H, D, F, CI, Br, I, OH, NH2, CN and optionally substituted C a alkyl; preferably selected from H, D, F, CI, Br, I, OH, NH2, CN and optionally substituted C 1-3 alkyl; preferably selected from H, D, F, CI, Br, I, OH, NH2, CN and optionally substituted C R 5 Selected from hydrogen, deuterium, halogen, hydroxyl, amino, cyano and C 1-6 Alkyl groups; preferably selected from H, D, F, Cl, Br, I, OH, NH2, CN, and C. 1-3 alkyl; T 1 Selected from -(CH2) n -, n is selected from 0, 1, and 2; T 2 Selected from -(CH2) m -, m is selected from 0, 1, 2 and 3; R a Selected from deuterium, halogen, hydroxyl, amino, cyano and C 1-6 Alkyl groups; preferably selected from D, F, Cl, Br, I, OH, NH2, CN and C. 1-3 alkyl.

2. The compound according to claim 1, wherein, Structural unit Selected from Preferred 3. The compound according to claim 1 or 2, wherein, The R 1 It is -Y-ONO2; Where Y is selected from C 1-10 Alkylene, C 3-12 Cycloalkylene, 3-12 membered heterocycloalkylene, C 6-10 aryl, 5-10 quinone heteroaryl, C 3-12 Cycloalkyl C 1-6 Alkylene, 3-12 membered heterocyclic alkylene C 1-6 Alkylene, C 6-10 Aromatic C 1-6 Alkylene, 5-10-membered heteroaryl C 1-6 Alkylene, C 1-10 Alkylene (C 1-6 (alkoxide) 1-3 C 3-12 Cycloalkyl C 1-6 alkeneoxy, 3-12 membered heterocyclic alkylene C 1-6 alkeneoxy, C 6-10 Aromatic C 1-6 alkeneoxy groups and 5-10 heteroaryl C groups 1-6 Alkyloxy; The groups described in Y are independently and arbitrarily selected by m1 R groups. c Replacement, m1 is selected from 0, 1, 2 and 3; R c Selected from deuterium, halogen, hydroxyl, amino, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Cyanoalkyl, C 1-6 Carboxyalkyl, C 1-6 Hydroxyalkyl and -(CH2) 0-6 -ONO2; preferably selected from D, F, Cl, Br, I, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Cyanoalkyl, C 1-3 Carboxyalkyl, C 1-3 Hydroxyalkyl and -(CH2) 0-3 -ONO2; The 3-12 membered heterocyclic alkyl group, 5-10 membered heteroaryl group, 3-12 membered heterocyclic alkyl group C 1-6 Alkylene, 3-12 membered heterocyclic alkylene C 1-6 alkeneoxy, 5-10 heteroaryl C 1-6 Alkylene and 5-10-membered heteroaryl C 1-6 Each alkoxide independently contains 1, 2, 3 or 4 heteroatoms or heterogroups independently selected from -NH-, -O-, -S- and N.

4. The compound according to any one of claims 1-3, having the structure shown in formula (I-1), or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of formula (I-1): R 2 R 3 R 4 R 5 And Y as defined in claim 3.

5. The compound according to any one of claims 1-4, having a structure as shown in formula (I-2) or (I-3), or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug having a structure as shown in formula (I-2) or (I-3): R 2 R 3 R 4 R 5 And Y as defined in claim 3.

6. The compound according to any one of claims 1-5, wherein, The R 2 and R 3 Each of the following is independently selected from H, D, F, Cl, Br, I, OH, and NH2; The R 4 Selected from H, D, F, Cl, Br, I, OH, NH2, CN, CH3 and CH2CH3; The R 5 Selected from H, D, F, Cl, Br, I, OH, NH2, CN and CH3; The R c Selected from D, F, Cl, Br, I, OH, NH2, CN, CH3, -CH2OH, -ONO2 and -CH2-ONO2.

7. The compound according to any one of claims 1-6, wherein, The Y is selected from C. 1-5 Alkylene, C 6-10 Alkylene, C 3-8 Monocycloalkyl substructure, C 5-10 Subbridged cycloalkyl, C 6-12 Spirocycloalkyl, 3-8 membered heterocycloalkyl, C 6-8 Alpha-aryl, C 3-8 Monocycloalkyl C 1-4 Alkylene, C 5-10 Subbridged cycloalkyl C 1-4 Alkylene, C 6-12 Spirocycloalkyl C 1-4 Alkylene, 3-8 quinone heterocyclic alkylene C 1-4 Alkylene, C 6-8 Aromatic C 1-4 Alkylene, C 1-6 Alkylene (C 1-4 (alkoxide) 1-3 C 3-8 Monocycloalkyl C 1-3 alkeneoxy, C 5-10 Subbridged cycloalkyl C 1-4 alkeneoxy, C 6-12 Spirocycloalkyl C 1-4 alkeneoxy group, 3-8 member heterocyclic alkyl group 14 alkeneoxy and C 6-8 Aromatic C 1-4 alkeneoxy group; the group described in Y is independently optionally occupied by m1 R c Replacement, m1 is selected from 0, 1, 2, and 3; R c Selected from D, F, Cl, Br, I, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 1-3 Cyanoalkyl, C 1-3 Carboxyalkyl, C 1-3 Hydroxyalkyl and -(CH2) 0-3 -ONO2; Preferably, Y is selected from -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)8-, and -(CH2). 10 -, -CH(CH3)-, -CH2-(OCH2CH2)3-, The R c Selected from D, F, Cl, Br, I, OH, NH2, CN, CH3, -CH2OH, -ONO2 and -CH2-ONO2; The m1 is selected from 0, 1, 2 and 3.

8. The compound according to claim 7, wherein, The Y is selected from -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)8-, and -(CH2) 10 -, -CH(CH3)-, -CH(CH3)CH2-, -C(CH3)2-, -C(CH3)2CH2-, -C(CH3)2(CH2)3-, -CH2-(OCH2CH2)3-, 9. A compound having the structure shown below, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

10. The compound according to claim 9, having the structure shown below, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

11. A pharmaceutical composition comprising a compound according to any one of claims 1-10, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, medium, or combination thereof.

12. Use of the compound according to any one of claims 1-10 or the pharmaceutical composition according to claim 11 in the preparation of a medicament for inhibiting ROCK kinase and / or myosin light chain phosphorylation.

13. The application according to claim 12, characterized in that, The drug that inhibits ROCK kinase and / or myosin light chain phosphorylation is a drug used to treat glaucoma and / or ocular hypertension.

14. A method for inhibiting ROCK kinase and / or myosin light chain phosphorylation, comprising contacting the cells with or administering to the subject a compound as claimed in any one of claims 1-10, or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound, or a pharmaceutical composition as claimed in claim 11; Preferably, the subject is a mammal, and more preferably a human.

15. A method of treating glaucoma and / or ocular hypertension in a patient, comprising administering to the patient a compound of any one of claims 1-10 or a stereoisomer, tautomer, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound or a pharmaceutical composition of claim 11.

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