SMTP compound and use thereof

Abstract

The present disclosure relates to an SMTP compound as shown in formula (I), a pharmaceutical composition containing the compound, and the use thereof in the prevention or treatment of cardiovascular and cerebrovascular diseases.

Description

SMTP compounds and uses thereof

[0001] Cross-reference to related applications

[0002] This application claims priority to and the benefit of Chinese Patent Application No. 202410478667.6, filed April 19, 2024, and Chinese Patent Application No. 202411164940.4, filed August 23, 2024, in the China National Intellectual Property Office, the disclosures of which are incorporated herein in their entireties by reference. TECHNICAL FIELD

[0003] The present disclosure relates to SMTP compounds, pharmaceutical compositions containing the compounds, and uses thereof in the prevention or treatment of thromboembolic diseases. BACKGROUND

[0004] The hemostatic system includes the coagulation system and the fibrinolytic system, which have important physiological functions in inhibiting bleeding and accelerating wound healing. Fibrinolysis is regulated by plasminogen and activated by physiological plasminogen activators, tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA). At the same time, the level of activated plasmin can be inhibited by blocking the activation of plasminogen through several specific molecules. However, various clinical cases have shown that genetic or acquired factors can enhance or weaken the fibrinolytic system, leading to disorders between the coagulation and fibrinolytic systems, resulting in bleeding or thrombosis.

[0005] Thrombus formation is much slower and less detectable than bleeding, leading to many patient deaths. Cerebral infarction is a major cause of death in developed countries. Even if life is saved, disability such as paralysis can still exist. Cerebral infarction can be treated by recanalization at the occlusion site, however, blood flow recanalization to the damaged site of cerebral infarction can cause hemorrhagic cerebral infarction and worsen life and functional prognosis. As a treatment method for recanalization of the occluded blood vessel in the acute phase of cerebral infarction, the currently known thrombolytic treatments are intravenous thrombolytic agents, rt-PA (recombinant tissue-type plasminogen activator), and intravascular surgical mechanical thrombectomy. The currently available drugs for treating thrombus include aspirin, ticlopidine, warfarin, and heparin, and rt-PA, which is the first choice drug, can be administered 4.5 hours after onset. However, this drug has a strong thrombolytic effect, but can cause hemorrhagic side effects, and there are also strict restrictions, allowing the proportion of patients to be administered to be less than 10% of all cerebral infarction patients. Even if rt-PA is applicable, a few percent of patients experience serious intracranial hemorrhagic side effects, and the risk of bleeding is a concern. Therefore, small molecules with different fibrinolytic mechanisms are ideal choices for new anti-thrombotic and thrombolytic drugs.

[0006] SMTPs (named after Stachybotrys microspora triprenyl phenols) is a novel family of small molecules produced by the fungus S. microspora. SMTP-7 is one of the SMTP family compounds with a broad spectrum of biological activities, including potentiation of the proteolytic activity of plasminogen, the circulating zymogen of plasmin, the major protease responsible for clot lysis. This activity of SMTP-7 can be attributed to the modulation of the plasminogen conformation: although resistant to proteolytic activation due to a helical, closed conformation, SMTP-7 relaxes the conformation to a conformation susceptible to activation. Thus, SMTP-7 promotes the physiological plasmin formation and clot clearance in vivo, effectively treating thrombotic and embolic strokes in rodent and primate models. Notably, the action of SMTP-7 is accompanied by a reduction in hemorrhagic transformation and a wider therapeutic time window. SMTP compounds are expected as therapeutic agents for thrombotic stroke, cytoprotective agents, etc. Given the huge unmet clinical need, the development of SMTP derivatives has broad application prospects. SUMMARY

[0007] The present disclosure relates to a compound of Formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0008] wherein, represents a single bond or a double bond;

[0009] a, b, c and d are each independently selected from a single bond or a double bond;

[0010] Q 1 and Q 2 are each independently selected from O or S;

[0011] R 1 is selected from COOH, -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9 , amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, tetrazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, C(O)N(C1-C6alkyl)2, or -5-10 membered heteroarylene-COOH, which amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, tetrazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, C(O)N(C1-C6alkyl)2, or -5-10 membered heteroarylene-COOH is optionally substituted with R 1a ;

[0012] R 1a selected from halogen, COOH, S(O)2OH, P(O)(OH)2, cyano, hydroxy, imidazolyl, tetrazolyl, OC(O)NHCi-C6alkyl, Ci-C6alkoxy, amino, NH(Ci-C6alkyl) or N(Ci-C6alkyl)2, said hydroxy, imidazolyl, tetrazolyl, OC(O)NHCi-C6alkyl, Ci-C6alkoxy, amino, NH(Ci-C6alkyl) or N(Ci-C6alkyl)2optionally substituted with R 1b ;

[0013] R 1b selected from COOH, S(O)2OH or P(O)(OH)2;

[0014] M is selected from O, NH or NR M ;

[0015] T is selected from (CH2) n , (CH2) n C(O)NH(CH2) m , phenylene, 4-12 membered heterocyclene or C3-C6cycloalkylene, said (CH2) n , (CH2) n C(O)NH(CH2) m , phenylene, 4-12 membered heterocyclene or C3-C6cycloalkylene optionally substituted with R T ;

[0016] R T selected from halogen, COOH, Ci-C6alkyl, amino or C3-C 10 cycloalkyl, said Ci-C6alkyl, amino or C3-C 10 cycloalkyl optionally substituted with R Ta ; or, two R T , together with their respective attached atoms, form a C3-C6cycloalkane ring or a 4-12 membered heterocyclic ring, said C3-C6cycloalkane ring or 4-12 membered heterocyclic ring optionally substituted with R 14 ;

[0017] R 14 selected from halogen, COOH, Ci-C6alkyl or amino;

[0018] R Ta selected from halogen, -S-Ci-C6alkyl, Ci-C6alkoxy, SH, hydroxy, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, COOH, imidazolyl, imidazolyl substituted with -SH, phenyl, indolyl, tetrazolyl, NHC(=NH)NH2or C(O)NH2;

[0019] R M selected from C1-C6 alkyl, said C1-C6 alkyl being optionally substituted with R Ma ; or R M and R T together with the atoms to which they are attached form a 4-12 membered heterocyclic ring, said 4-12 membered heterocyclic ring being optionally substituted with R 13 ;

[0020] R 13 selected from hydroxyl, cyano, SH, amino, carboxyl, halogen or C1-C6 alkyl;

[0021] R Ma selected from COOH, hydroxyl or

[0022] n is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0023] m is selected from 1, 2, 3, 4, 5, 6, 7 or 8;

[0024] R 2 and R 4 are each independently selected from OH, SH, amino, C1-C6 alkoxy, -OC(O)N(C1-C6 alkyl)2, -OC(O)NH(C1-C6 alkyl), -OC(O)NH2 or OC(O)C1-C6 alkyl; said amino, C1-C6 alkoxy, -OC(O)N(C1-C6 alkyl)2, -OC(O)NH(C1-C6 alkyl), -OC(O)NH2 or OC(O)C1-C6 alkyl being optionally substituted with R 2a ;

[0025] R 2a selected from halogen, amino, COOH, NHC1-C6 alkyl or N(C1-C6 alkyl)2;

[0026] R 3 and R 5 are each independently selected from hydrogen, halogen, OH, SH, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, OC(O)C1-C6 alkyl or OC(O)H;

[0027] R 6 selected from hydrogen or C1-C6 alkyl; said C1-C6 alkyl being optionally substituted with R 6a ;

[0028] R 6a selected from halogen;

[0029] R 7 and R 8each independently selected from hydrogen, halogen, hydroxyl, cyano, carboxyl, Ci-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-10 membered heteroaryl, said Ci-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-10 membered heteroaryl being optionally substituted with R 10 cycloalkyl, C2-C6alkenyl, or 5-10 membered heteroaryl, said Ci-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-10 membered heteroaryl being optionally substituted with R 10 cycloalkyl, C2-C6alkenyl, or 5-10 membered heteroaryl, said Ci-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-10 membered heteroaryl being optionally substituted with R 7a substituted;

[0030] R 7a selected from amino, halogen, hydroxyl, NH(Ci-C6alkyl), or N(Ci-C6alkyl)2;

[0031] R 9 selected from hydroxyl, COOH, P(O)(OH)2, S(O)2OH, S(O)OH, imidazolyl, or tetrazolyl;

[0032] R 11 selected from hydrogen, deuterium, halogen, or Ci-C6alkyl, said Ci-C6alkyl being optionally substituted with R 11a substituted; or R 11 , R 5 and the carbon atom to which they are attached together form C=CH2or C=NOH;

[0033] R 12 selected from hydrogen, deuterium, halogen, or Ci-C6alkyl, said Ci-C6alkyl being optionally substituted with R 12a substituted; or R 12 , R 3 and the carbon atom to which they are attached together form C=CH2or C=NOH;

[0034] R 11a and R 12a each independently selected from halogen

[0035] R 15 and R 16 each independently selected from hydrogen or deuterium.

[0036] In some embodiments,

[0037] T is selected from (CH2) n , phenylene, 4-12 membered heterocyclene, or C3-C6cycloalkylene, said (CH2) n , phenylene, 4-12 membered heterocyclene, or C3-C6cycloalkylene being optionally substituted with R T substituted;

[0038] R Taselected from halogen, -S-Ci-C6alkyl, Ci-C6alkoxy, SH, hydroxy, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, COOH, imidazolyl, tetrazolyl, NHC(=NH)NH2, or C(O)NH2;

[0039] R 9 selected from hydroxy, COOH, P(O)(OH)2, S(O)2OH, imidazolyl, or tetrazolyl.

[0040] In some embodiments, is a double bond, and a, b, c, and d are each a single bond.

[0041] In some embodiments, is a double bond, and a, b, c, and d are each a single bond.

[0042] In some embodiments, a, b, c, and d are each a double bond, or a, b, c, and d are each a single bond.

[0043] In some embodiments, Q 1 and Q 2 are each O.

[0044] In some embodiments, R 1 is selected from COOH, -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9 , amino, NHCi-C6alkyl, N(Ci-C6alkyl)2, CH=N(Ci-C6alkyl), imidazolyl, tetrazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)Ci-C6alkyl, Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2, which amino, NHCi-C6alkyl, N(Ci-C6alkyl)2, CH=N(Ci-C6alkyl), imidazolyl, tetrazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)Ci-C6alkyl, Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2is optionally substituted with R 1a .

[0045] In some embodiments, R 1 is selected from -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9, amino, NHCi-C6alkyl, N(Ci-C6alkyl)2, CH=N(Ci-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)Ci-C6alkyl, Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2, said amino, NHCi-C6alkyl, N(Ci-C6alkyl)2, CH=N(Ci-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)Ci-C6alkyl, Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2optionally substituted with R 1a substituted.

[0046] In some embodiments, R 1 is selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , amino, NHCi-C6alkyl, N(Ci-C6alkyl)2, CH=N(Ci-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)Ci-C6alkyl, Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2, said amino, NHCi-C6alkyl, N(Ci-C6alkyl)2, CH=N(Ci-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)Ci-C6alkyl, Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2optionally substituted with R 1a substituted.

[0047] In some embodiments, R 1 is selected from COOH, -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2, said Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2optionally substituted with R 1a substituted.

[0048] In some embodiments, R 1 is selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2, said Ci-C6alkyl, C2-C6alkenyl, or C(O)N(Ci-C6alkyl)2optionally substituted with R 1a substituted.

[0049] In some embodiments, R 1 selected from COOH, -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , C1-C3 alkyl, C2-C4 alkenyl, C(O)N(C1-C3 alkyl)2, or -5-6 membered heteroarylene-COOH, said C1-C3 alkyl, C2-C4 alkenyl, C(O)N(C1-C3 alkyl)2, or -5-6 membered heteroarylene-COOH optionally substituted with R 1a In some embodiments, R 1 selected from -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9 , CH=N(C1-C6)alkyl, C1-C6 alkyl, C2-C6 alkenyl, or C(O)N(C1-C6 alkyl)2, said CH(=N)C1-C6 alkyl, C1-C6 alkyl, C2-C6 alkenyl, or C(O)N(C1-C6 alkyl)2 optionally substituted with R 1a .

[0050] In some embodiments, R 1 selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , C1-C3 alkyl, C2-C4 alkenyl, or -5-6 membered heteroarylene-COOH, said C1-C3 alkyl, C2-C4 alkenyl, or -5-6 membered heteroarylene-COOH optionally substituted with R 1a .

[0051] In some embodiments, R 1 selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , C1-C3 alkyl, or C2-C4 alkenyl, said C1-C3 alkyl, or C2-C4 alkenyl optionally substituted with R 1a .

[0052] In some embodiments, R 1a selected from COOH, S(O)2OH, P(O)(OH)2, hydroxyl, imidazolyl, tetrazolyl, C1-C6 alkoxy, amino, NH(C1-C6 alkyl), or N(C1-C6 alkyl)2, said C1-C6 alkoxy, amino, NH(C1-C6 alkyl), or N(C1-C6 alkyl)2 optionally substituted with R 1b .

[0053] In some embodiments, R 1aselected from COOH, hydroxyl, C1-C3alkoxy, NH(C1-C3alkyl), or OC(O)NHC1-C3alkyl, said C1-C3alkoxy, NH(C1-C3alkyl), or OC(O)NHC1-C3alkyl being optionally substituted with R 1b substituted.

[0054] In some embodiments, R 1a is selected from COOH, hydroxyl, C1-C3alkoxy, or NH(C1-C3alkyl), said C1-C3alkoxy or NH(C1-C3alkyl) being optionally substituted with R 1b substituted.

[0055] In some embodiments, R 1b is selected from COOH.

[0056] In some embodiments, M is selected from O, NH, or NR M ; R M is selected from C1-C6alkyl, said C1-C6alkyl being optionally substituted with R Ma ; or R M and R T , together with the atom to which they are attached, form a 4-10 membered heterocyclic ring, said 4-10 membered heterocyclic ring being optionally substituted with R 13 .

[0057] In some embodiments, M is selected from O, NH, or NR M ; R M is selected from C1-C3alkyl, said C1-C3alkyl being optionally substituted with R Ma ; or R M and R T , together with the atom to which they are attached, form a 4-6 membered heterocyclic ring, said 4-6 membered heterocyclic ring being optionally substituted with R 13 .

[0058] In some embodiments, M is selected from O, NH, or NR M ; R M is selected from C1-C3alkyl, said C1-C3alkyl being optionally substituted with R Ma ; or R M and R T , together with the atom to which they are attached, form a 4 membered heterocyclic ring, said 4 membered heterocyclic ring being optionally substituted with R 13 .

[0059] In some embodiments, M is selected from O, NH, or NR M ; R M is selected from methyl, said methyl being optionally substituted with R Ma ; or R M and R Tand the atoms to which they are attached form an aziridinylene, thiazolidinylene, pyrrolidinylene, piperazinylene, or morpholinylene group, which is optionally substituted with R 13 substituted.

[0060] In some embodiments, R Ma is selected from COOH or hydroxyl.

[0061] In some embodiments, R Ma is selected from COOH or

[0062] In some embodiments, R Ma is selected from COOH.

[0063] In some embodiments, R 13 is selected from halogen or C1-C6 alkyl.

[0064] In some embodiments, R 13 is selected from hydroxyl, halogen, or C1-C3 alkyl.

[0065] In some embodiments, R 13 is selected from F, hydroxyl, or methyl.

[0066] In some embodiments, M is selected from O or NH.

[0067] In some embodiments, M is selected from O.

[0068] In some embodiments, M is selected from NH.

[0069] In some embodiments, M is selected from NH or NR M .

[0070] In some embodiments, T is selected from (CH2) n , phenylene, 4-12 membered heterocyclidene, or C3-C6 cycloalkyidene, which (CH2) n , phenylene, 4-12 membered heterocyclidene, or C3-C6 cycloalkyidene is optionally substituted with R T .

[0071] In some embodiments, T is selected from (CH2) n , phenylene, or 4-12 membered heterocyclidene, which (CH2) n , phenylene, or 4-12 membered heterocyclidene is optionally substituted with R T .

[0072] In some embodiments, T is selected from (CH2) n , phenylene, or 4-10 membered heterocyclidene, which (CH2)n , phenylene or 4-10 membered heterocyclylene, optionally substituted with R T .

[0073] In some embodiments, T is selected from (CH2) n , phenylene or C4-C5 cycloalkylene, optionally substituted with R n , phenylene or C4-C5 cycloalkylene, optionally substituted with R T .

[0074] In some embodiments, T is selected from (CH2) n , phenylene or 4-6 membered heterocyclylene, optionally substituted with R n , phenylene or 4-6 membered heterocyclylene, optionally substituted with R T .

[0075] In some embodiments, T is selected from (CH2) n , (CH2) n C(O)NH(CH2) m , phenylene, 4-6 membered heterocyclylene or C3-C6 cycloalkylene, optionally substituted with R n , (CH2) n C(O)NH(CH2) m , phenylene, 4-6 membered heterocyclylene or C3-C6 cycloalkylene, optionally substituted with R T .

[0076] In some embodiments, T is selected from (CH2) n , optionally substituted with R n . T

[0077] In some embodiments, n is selected from 0, 1, 2 or 3.

[0078] In some embodiments, n is selected from 0, 1 or 2.

[0079] In some embodiments, n is selected from 1 or 2.

[0080] In some embodiments, m is selected from 1 or 2.

[0081] In some embodiments, T is selected from (CH2) n C(O)NH(CH2) m , wherein n is selected from 1 or 2, and m is 1.

[0082] In some embodiments, R T is selected from halogen, COOH, C1-C6 alkyl or amino, optionally substituted with R Ta ; or, two R​T and the atoms of each to which they are attached form, together with the carbon atom to which they are attached, a C3-C6cycloalkane ring or a 4-12 membered heterocyclic ring, which is optionally substituted with R 14 .

[0083] In some embodiments, R T is selected from amino, COOH, or C1-C6alkyl, which is optionally substituted with R Ta ; or, two R T and the atoms of each to which they are attached form, together with the carbon atom to which they are attached, a C3-C6cycloalkane ring, which is optionally substituted with R 14 ; or R M and R T and the atoms of each to which they are attached form, together with the carbon atom to which they are attached, a 4-10 membered heterocyclic ring, which is optionally substituted with R 13 .

[0084] In some embodiments, R T is selected from amino, COOH, C1-C3alkyl, or C3-C6cycloalkyl, which is optionally substituted with R Ta ; or, two R T and the atoms of each to which they are attached form, together with the carbon atom to which they are attached, a C3-C4cycloalkane ring or a 4-6 membered heterocyclic ring, which is optionally substituted with R 14 ; or R M and R T and the atoms of each to which they are attached form, together with the carbon atom to which they are attached, a 4-6 membered heterocyclic ring, which is optionally substituted with R 13 .

[0085] In some embodiments, R T is selected from amino, COOH, or C1-C3alkyl, which is optionally substituted with R Ta ; or, two R T and the atoms of each to which they are attached form, together with the carbon atom to which they are attached, a C3-C4cycloalkane ring; or R M and R T and the atoms of each to which they are attached form, together with the carbon atom to which they are attached, a 4 membered heterocyclic ring.

[0086] In some embodiments, R T is selected from amino, COOH, methyl, or which is optionally substituted with R ; or, two R Ta are taken together with the carbon atom to which they are attached to form a 4-10 membered heterocyclic ring, which is optionally substituted with R Tand the atoms to which they are attached together form a cyclopropyl, cyclobutyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperazinyl or thietanyl group, wherein the cyclopropyl, cyclobutyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperazinyl or thietanyl group is optionally replaced by R 14 replace.

[0087] In some embodiments, R T is selected from halogen, COOH, C1-C6 alkyl, amino or C3-C6 cycloalkyl, wherein the C1-C6 alkyl, amino or C3-C6 cycloalkyl is optionally replaced by R Ta Replace; or, two R T and their respective atoms together form a C3-C6 cycloalkane ring or a 4-6 membered heterocyclic ring, wherein the C3-C6 cycloalkane ring or the 4-6 membered heterocyclic ring is optionally replaced by R 14 Replacement; or R M With R T and the atoms to which they are attached together form a 4-6 membered heterocyclic ring, wherein the 4-6 membered heterocyclic ring is optionally replaced by R 13 replace.

[0088] In some embodiments, R Ta Selected from halogen, -S-C1-C6 alkyl, C1-C6 alkoxy, SH, hydroxy, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COOH, imidazolyl, tetrazolyl, NHC(=NH)NH2 or C(O)NH2.

[0089] In some embodiments, R Ta is selected from halogen, -S-C1-C6 alkyl, SH, hydroxy, amino, COOH, imidazolyl, imidazolyl substituted by -SH, phenyl, indolyl, NHC(=NH)NH2 or C(O)NH2.

[0090] In some embodiments, R Ta is selected from F, SCH3, SH, hydroxy, amino, COOH, 2-imidazolyl, 2-SH-4-imidazolyl, phenyl, 3-indolyl, NHC(=NH)NH2 or C(O)NH2.

[0091] In some embodiments, R Ta Selected from SH, hydroxy, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COOH, imidazolyl, tetrazolyl, NHC(=NH)NH2 or C(O)NH2.

[0092] In some embodiments, R Tais selected from halogen, -S-C1-C3 alkyl, SH, hydroxy, amino, COOH, imidazolyl, NHC(=NH)NH2 or C(O)NH2.

[0093] In some embodiments, R Ta is selected from SH, hydroxy, amino, COOH, imidazolyl, NHC(=NH)NH2 or C(O)NH2.

[0094] In some embodiments, R T Selected from C1-C6 alkyl or amino; or, two R T and the atoms to which they are attached together form a C3-C6 cycloalkane ring.

[0095] In some embodiments, R 14 Selected from C1-C6 alkyl.

[0096] In some embodiments, R 14 Selected from C1-C3 alkyl.

[0097] In some embodiments, R 9 Selected from COOH, P(O)(OH)2, S(O)2OH, imidazolyl or tetrazolyl.

[0098] In some embodiments, R 9 Selected from hydroxy, COOH, P(O)(OH)2, tetrazolyl or S(O)2OH.

[0099] In some embodiments, R 9 Selected from COOH, P(O)(OH)2 or tetrazolyl.

[0100] In some embodiments, R 9 Selected from COOH.

[0101] In some embodiments, R 1 Selected from C(O)MTR 9 .

[0102] In some embodiments, M is selected from O, NH, or NR M , R M With R T and the atoms to which they are attached together form a 4-6 membered heterocyclic ring, wherein the 4-6 membered heterocyclic ring is optionally replaced by R 13 replace;

[0103] T is selected from (CH2) n , the (CH2) n Optional R T Replacement; R T is selected from COOH or C1-C3 alkyl, wherein the C1-C3 alkyl is optionally replaced by RTa Replace; or, two R T and their respective atoms together form a C3-C6 cycloalkane ring, the C3-C6 cycloalkane ring being optionally replaced by R 14 substituted; n is selected from 0, 1 or 2;

[0104] R Ta is selected from halogen, -S-C1-C3 alkyl, SH, hydroxy, amino or COOH;

[0105] R 9 Selected from COOH.

[0106] In some embodiments, M is selected from O, NH, or NR M , R M With R T and the atoms to which they are attached together form a 5-membered heterocyclic ring;

[0107] T is selected from (CH2) n , the (CH2) n Optional R T Replacement; R T is selected from COOH or C1-C3 alkyl, wherein the C1-C3 alkyl is optionally replaced by R Ta Replace; or, two R T and their respective atoms together form a C3-C4 cycloalkane ring; n is selected from 1;

[0108] R Ta selected from halogen;

[0109] R 9 Selected from COOH.

[0110] In some embodiments, M is selected from O, NH, or NR M , R M With R T and the atoms to which it is attached together form a thiazolylidene group;

[0111] T is selected from CH2, said CH2 being optionally replaced by R T Replacement; R T is selected from COOH or methyl, the methyl group is optionally replaced by R Ta Replace; or, two R T and the atoms to which they are attached together form a cyclopropyl group;

[0112] R Ta selected from halogen;

[0113] R 9 Selected from COOH.

[0114] In some embodiments, R 1 Selected from COOH,

[0115] In some embodiments, R 1 is selected from

[0116] In some embodiments, R 1 is selected from

[0117] In some embodiments, R 1 is selected from C(O)M-T-R 9 .

[0118] In some embodiments, R 1 is selected from C(O)M(CH2) n R 9 wherein M is selected from O, NH, NCH3, or NCH2COOH; n is 0, 1, 2, 3, 4, 5, or 6; R 9 is selected from hydroxyl, COOH, P(O)(OH)2, S(O)2OH, S(O)OH, or tetrazolyl; said (CH2) n is optionally substituted with 1, 2, 3, or 4 substituents independently selected from halogen, COOH, C1-C6 alkyl, amino, or C3-C 10 cycloalkyl, said C1-C6 alkyl, amino, or C3-C 10 cycloalkyl is optionally substituted with 1, 2, or 3 substituents independently selected from halogen, -S-C1-C6 alkyl, SH, hydroxyl, amino, COOH, imidazolyl, imidazolyl substituted with -SH, phenyl, indolyl, NHC(=NH)NH2, or C(O)NH2.

[0119] In some embodiments, R 1 is selected from C(O)M(CH2) n R 9 wherein M is selected from O, NH, or NCH3; n is 0, 1, 2, or 3; R 9 is selected from hydroxyl, COOH, P(O)(OH)2, S(O)2OH, S(O)OH, or tetrazolyl; said (CH2) noptionally substituted with 1 or 2 substituents independently selected from halogen, COOH, C1-C6alkyl, amino, or C3-C6cycloalkyl, said C1-C6alkyl, amino, or C3-C6cycloalkyl optionally substituted with 1, 2, or 3 substituents independently selected from halogen, -S-C1-C6alkyl, SH, hydroxyl, amino, COOH, imidazolyl, imidazolyl substituted with -SH, phenyl, indolyl, NHC(=NH)NH2, or C(O)NH2.

[0120] In some embodiments, R 1 is selected from C(O)M(CH2) n R 9 wherein M is NH; n is 1 or 2; R 9 is selected from COOH, P(O)(OH)2, or S(O)2OH; said (CH2) n optionally substituted with 1 or 2 substituents independently selected from halogen or C1-C6alkyl, said C1-C6alkyl optionally substituted with 1, 2, or 3 substituents independently selected from halogen, -S-C1-C3alkyl, SH, hydroxyl, amino, COOH, 2-imidazolyl, 4-imidazolyl substituted with -SH, phenyl, 3-indolyl, NHC(=NH)NH2, or C(O)NH2.

[0121] In some embodiments, R 1 is selected from

[0122] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is O or NH; T is phenylene, 4-12 membered heterocyclene, or C3-C6cycloalkylene, said phenylene, 4-12 membered heterocyclene, or C3-C6cycloalkylene optionally substituted with 1 or 2 substituents selected from halogen or C1-C6alkyl; R 9 is selected from COOH, P(O)(OH)2, or S(O)2OH.

[0123] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is O or NH; T is phenylene, 4-6 membered heterocyclene, or C3-C6cycloalkylene, said phenylene, 4-6 membered heterocyclene, or C3-C6cycloalkylene optionally substituted with 1 or 2 substituents selected from halogen or C1-C3alkyl; R 9 is COOH.

[0124] In some embodiments, R 1 is selected from

[0125] In some embodiments, R 1 is selected from

[0126] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is NR M , T is (CH2) T substituted with R n , n is 1, 2 or 3, R M and R T together with the atoms to which they are attached form a 4-12 membered heterocyclic ring, said 4-12 membered heterocyclic ring being optionally substituted with 1 or 2 substituents independently selected from halogen, OH or C1-C6 alkyl, R 9 is selected from COOH, P(O)(OH)2 or S(O)2OH.

[0127] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is NR M , T is (CH2) T substituted with R n , n is 1 or 2, R M and R T together with the atoms to which they are attached form a 4-6 membered heterocyclic ring, said 4-6 membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O or S, and being optionally substituted with 1 or 2 substituents independently selected from halogen, OH or C1-C6 alkyl, R 9 is selected from COOH.

[0128] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is NR M , T is (CH2) T substituted with R n , n is 1 or 2, R M and R T together with the atoms to which they are attached form an azetidine, piperazine, thiazolidine, pyrrolidine or morpholine, said azetidine, piperazine, thiazolidine, pyrrolidine or morpholine being optionally substituted with 1 or 2 substituents independently selected from halogen, OH or C1-C3 alkyl, R 9 is selected from COOH.

[0129] In some embodiments, R 1 is selected from

[0130] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is O or NH, T is C(R T )2or CH2C(R T )2, and two R T groups and the C atoms to which they are attached together form a C3-C6 cycloalkane ring or a 4-12 membered heterocyclic ring, which is optionally substituted with 1 or 2 substituents independently selected from halogen and C1-C6 alkyl, R 9 is selected from COOH, P(O)(OH)2, or S(O)2OH.

[0131] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is O or NH, T is C(R T )2or CH2C(R T )2, and two R T groups and the C atoms to which they are attached together form a C3-C6 cycloalkane ring or a 4-6 membered heterocyclic ring, which is optionally substituted with 1 or 2 substituents independently selected from halogen and C1-C6 alkyl, R 9 is selected from COOH.

[0132] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is O or NH, T is C(R T )2or CH2C(R T )2, and two R T groups and the C atoms to which they are attached together form a C3-C6 cycloalkane ring or a 4-6 membered heterocyclic ring, which is optionally substituted with 1 or 2 substituents independently selected from halogen and C1-C3 alkyl, R 9 is selected from COOH.

[0133] In some embodiments, R 1 is selected from C(O)M-T-R 9 wherein M is O or NH, T is C(R T )2or CH2C(R T )2, and two R T groups and the C atoms to which they are attached together form a cyclopropane, cyclobutane, azetidine, oxetane, tetrahydrofuran, tetrahydropyran, or piperidine, which is optionally substituted with 1 or 2 substituents independently selected from F, CH3, or C2H5, R 9 is selected from COOH.

[0134] In some embodiments, R 1 is selected from

[0135] In some embodiments, R 1 is selected from C(O)M(CH2) n C(O)NH(CH2) m R 9 wherein M is selected from O, NH or NCH3, n is selected from 1, 2, 3, 4, 5 or 6, m is selected from 1, 2, 3, 4, 5 or 6, R 9 is selected from hydroxyl, COOH, P(O)(OH)2, S(O)2OH, S(O)OH or tetrazolyl; said (CH2) n C(O)NH(CH2) m is optionally substituted with 1, 2, 3 or 4 substituents independently selected from halogen or C1-C6alkyl optionally substituted with 1, 2 or 3 substituents independently selected from halogen, hydroxyl, amino or imidazolyl.

[0136] In some embodiments, R 1 is selected from C(O)M(CH2) n C(O)NH(CH2) m R 9 wherein M is NH, n is selected from 1 or 2, m is 1 or 2, R 9 is selected from COOH; said (CH2) n C(O)NH(CH2) m is optionally substituted with 1 or 2 substituents independently selected from halogen or C1-C3alkyl optionally substituted with 1, 2 or 3 substituents independently selected from halogen, hydroxyl, amino or 4-imidazolyl.

[0137] In some embodiments, R 1 is selected from

[0138] In some embodiments, R 1 is selected from

[0139] In some embodiments, R 1 is selected from 5-6 membered heteroarylene-COOH.

[0140] In some embodiments, R 1 is selected from

[0141] In some embodiments, with R 1 The connected C atoms are in S configuration.

[0142] In some embodiments, R 2 and R 4 Each independently selected from SH, amino, C1-C6 alkoxy, -OC(O)N(C1-C6 alkyl)2, -OC(O)NH(C1-C6 alkyl), -OC(O)NH2 or OC(O)C1-C6 alkyl; the amino, C1-C6 alkoxy, -OC(O)N(C1-C6 alkyl)2, -OC(O)NH(C1-C6 alkyl), -OC(O)NH2 or OC(O)C1-C6 alkyl are optionally replaced by R 2a replace.

[0143] In some embodiments, R 2 and R 4 Each independently selected from SH, amino, C1-C3 alkoxy or OC(O)C1-C3 alkyl, wherein the C1-C3 alkoxy, OC(O)C1-C3 alkyl is optionally replaced by R 2a replace.

[0144] In some embodiments, R 2a Selected from halogen, COOH or N(C1-C3 alkyl)2.

[0145] In some embodiments, R 2 and R 4 Each independently selected from OH, SH, amino, C1-C6 alkoxy or OC(O)C1-C6 alkyl, wherein the amino, C1-C6 alkoxy or OC(O)C1-C6 alkyl is optionally replaced by R 2a replace.

[0146] In some embodiments, R 2 and R 4 Each is independently selected from OH, SH, amino, C1-C3 alkoxy or OC(O)C1-C3 alkyl, wherein the amino, C1-C3 alkoxy or OC(O)C1-C3 alkyl is optionally substituted with 1 or 2 substituents selected from halogen, COOH or N(C1-C3 alkyl)2.

[0147] In some embodiments, R 2 and R 4 Each independently selected from OH, SH, amino, OCHF2,

[0148] In some embodiments, R 2 and R 4 are each independently selected from OH. In some embodiments, R2 and R 4 are the same.

[0149] In some embodiments, R 3 and R 5 are each independently selected from hydrogen, halogen, SH, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, OC(0)Ci-C6alkyl, or OC(0)H.

[0150] In some embodiments, R 3 and R 5 are each independently selected from amino, OC(0)Ci-C3alkyl, or OC(0)H.

[0151] In some embodiments, R 3 and R 5 are each independently selected from OH.

[0152] In some embodiments, R 3 and R 5 are each independently selected from OH, amino, OC(0)Ci-C6alkyl, or OC(0)H.

[0153] In some embodiments, R 3 and R 5 are each independently selected from OH, amino, OC(0)CH3, or OC(0)H.

[0154] In some embodiments, R 3 and R 5 are the same.

[0155] In some embodiments, the C atom attached to R 3 is in the S configuration.

[0156] In some embodiments, the C atom attached to R 5 is in the S configuration.

[0157] In some embodiments, the C atoms attached to R 3 and R 5 are each in the S configuration.

[0158] In some embodiments, R 6 is selected from hydrogen or Ci-C6alkyl.

[0159] In some embodiments, R 6 is selected from hydrogen or methyl.

[0160] In some embodiments, R 6 is selected from Ci-C3alkyl.

[0161] In some embodiments, R6 Selected from hydrogen.

[0162] In some embodiments, R 7 and R 8 Each independently selected from hydrogen, halogen, hydroxyl, C1-C6 alkyl or C3-C 10 Cycloalkyl, the C1-C6 alkyl or C3-C 10 The cycloalkyl group is optionally replaced by R 7a replace.

[0163] In some embodiments, R 7 and R 8 Each independently selected from halogen, hydroxy, C1-C6 alkyl or C3-C6 cycloalkyl, the C1-C6 alkyl or C3-C6 cycloalkyl optionally replaced by R 7a replace.

[0164] In some embodiments, R 7 and R 8 Each is independently selected from halogen, cyano, carboxyl, C1-C3 alkyl, C3-C6 cycloalkyl, C2-C4 alkenyl or 5-membered heteroaryl, wherein the C1-C3 alkyl, C3-C6 cycloalkyl, C2-C4 alkenyl or 5-membered heteroaryl is optionally replaced by R 7a replace.

[0165] In some embodiments, R 7 and R 8 Each independently selected from halogen, C1-C3 alkyl or C3-C6 cycloalkyl, the C1-C3 alkyl is optionally replaced by R 7a replace.

[0166] In some embodiments, R 7a Selected from amino, NH(C1-C6 alkyl) or N(C1-C6 alkyl)2.

[0167] In some embodiments, R 7a Selected from amino, NH(C1-C3 alkyl) or N(C1-C3 alkyl)2.

[0168] In some embodiments, R 7a is selected from halogen, hydroxyl or N(C1-C3 alkyl)2. In some embodiments, R 7a Selected from fluorine, hydroxyl or N(CH3)2.

[0169] In some embodiments, R 7a Selected from N(C1-C3 alkyl)2.

[0170] In some embodiments, R 7 and R 8 are each independently selected from hydrogen.

[0171] In some embodiments, R 7 and R 8 are each independently selected from hydrogen, halogen, cyano, carboxyl, C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-6 membered heteroaryl, said C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-6 membered heteroaryl optionally substituted with R 7a .

[0172] In some embodiments, R 7 and R 8 are each independently selected from hydrogen, halogen, cyano, carboxyl, C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl, said C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl optionally substituted with 1, 2, or 3 substituents independently selected from halogen, OH, and N(C1-C3alkyl)2.

[0173] In some embodiments, R 7 and R 8 are each independently selected from hydrogen, bromine, cyano, methyl, CF3, CHF2, CH2OH, CH2N(CH3)2, vinyl, COOH, cyclopropyl, or pyrazolyl.

[0174] In some embodiments, R 7 and R 8 are the same.

[0175] In some embodiments, R 11 is selected from deuterium, halogen, or C1-C6alkyl, said C1-C6alkyl optionally substituted with R 11a ; or R 11 , R 5 and the carbon atom to which they are attached together form C=CH2or C=NOH.

[0176] In some embodiments, R 11 , R 5 and the carbon atom to which they are attached together form C=NOH.

[0177] In some embodiments, R 11 is selected from hydrogen.

[0178] In some embodiments, R 11 is selected from hydrogen; or R 11 , R 5 and the carbon atom to which they are attached together form C=NOH.

[0179] In some embodiments, R 12 is selected from deuterium, halogen, or C1-C6alkyl, said C1-C6alkyl optionally substituted with R12a substituted; or R 12 , R 3 and the carbon atom to which they are attached together form C=CH2or C=NOH.

[0180] In some embodiments, R 12 , R 3 and the carbon atom to which they are attached together form C=NOH.

[0181] In some embodiments, R 12 is selected from hydrogen.

[0182] In some embodiments, R 12 is selected from hydrogen, or R 12 , R 3 and the carbon atom to which they are attached together form C=NOH.

[0183] In some embodiments, R 3 and R 5 are the same, and R 11 and R 12 are the same.

[0184] In some embodiments, R 11 , R 5 and the carbon atom to which they are attached together form C=NOH, and R 12 , R 3 and the carbon atom to which they are attached together form C=NOH.

[0185] In some embodiments, R 1 is COOH, and R 6 is methyl.

[0186] In some embodiments, R 1 is COOH, and R 3 and R 5 are the same, both being amino, OC(O)Ci-C6alkyl or OC(O)H. In some embodiments, R 1 is COOH, and R 3 and R 5 are the same, both being OH, amino, OC(O)CH3or OC(O)H.

[0187] In some embodiments, R 1 is COOH, and a, b, c and d are each single bonds.

[0188] In some embodiments, R 1 is COOH, and R 7 and R 8are the same and are each hydrogen, halogen, cyano, carboxyl, C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl, said C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl optionally substituted with 1, 2, or 3 substituents independently selected from halogen, OH, and N(C1-C3alkyl)2.

[0189] In some embodiments, R 1 is COOH, and R 7 and R 8 are the same and are each hydrogen, bromine, cyano, methyl, CF3, CHF2, CH2OH, CH2N(CH3)2, vinyl, COOH, cyclopropyl, or pyrazolyl.

[0190] In some embodiments, R 1 is COOH, and R 2 and R 4 are the same and are each OH, SH, amino, C1-C3alkoxy, or OC(O)C1-C3alkyl, said amino, C1-C3alkoxy, or OC(O)C1-C3alkyl optionally substituted with 1 or 2 substituents selected from halogen, COOH, or N(C1-C3alkyl)2.

[0191] In some embodiments, R 1 is COOH, and R 2 and R 4 are the same and are each SH, amino, OCHF2,

[0192] In some embodiments, R 2 and R 4 are the same and are each OH; R 3 and R 5 are the same and are each OH; R 11 and R 12 are the same and are each H; R 7 and R 8 are the same and are each H; and / or R 6 is H.

[0193] In some embodiments, R 15 and R 16 are each hydrogen.

[0194] In some embodiments, R 15 and R 16 are each deuterium.

[0195] In some embodiments, the compound of Formula (I) described herein is selected from the group consisting of compounds of Formula (I’):

[0196] wherein a, b, c, d, Q 1 , Q 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 11 or R 12 are as defined above. In some embodiments, the compound of Formula (I) according to the present disclosure is selected from a compound of Formula (II):

[0197] wherein a, b, c, d, Q 1 , Q 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 11 or R 12 are as defined above.

[0198] In some embodiments, the compound of Formula (I) according to the present disclosure is selected from a compound of Formula (III):

[0199] wherein a, b, c, d, Q 1 , Q 2 , M, T, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 11 or R 12 are as defined above.

[0200] In another aspect, the present disclosure provides the following compounds, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0201] In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of Formula (I) of the present disclosure, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0202] In another aspect, the present disclosure provides a method of treating cardiovascular and cerebrovascular diseases in a mammal, comprising administering to a mammal, preferably a human, in need of such treatment, a therapeutically effective amount of a compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0203] In another aspect, the present disclosure provides a use of a compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating cardiovascular and cerebrovascular diseases.

[0204] In another aspect, the present disclosure provides a use of a compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in preventing or treating cardiovascular and cerebrovascular diseases.

[0205] In another aspect, the present disclosure provides a compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating cardiovascular and cerebrovascular diseases.

[0206] In some embodiments, the cardiovascular and cerebrovascular diseases are selected from thromboembolic diseases.

[0207] In some embodiments, the cardiovascular and cerebrovascular diseases are selected from acute ischemic stroke.

[0208] The compounds and pharmaceutical compositions of the present application have one or more of the following beneficial effects:

[0209] The compounds and pharmaceutical compositions of the present application have one or more of the following beneficial effects: (1) significant profibrinolytic activity, which can rapidly increase the level of plasmin and promote its thrombus dissolution; (2) good antioxidant activity; (3) good anti-inflammatory activity; (4) high drug exposure (AUC) and good PK properties; (5) possible realization of clinical intravenous bolus administration, greatly improving the convenience of administration and reducing the risk of stroke progression during inter-hospital transport; (6) improvement of cerebral infarction after cerebral infarction, which can reduce the risk of cerebral hemorrhage; (7) lower blood toxicity.

[0210] Definitions and explanations of terms

[0211] Unless otherwise indicated, the terms used in the present disclosure have the following meanings, the definitions of groups and terms recited in the present disclosure, including the definitions as examples, exemplary definitions, preferred definitions, definitions recited in tables, definitions of specific compounds in examples, etc., can be combined and integrated with each other arbitrarily. A particular term should not be considered indefinite or unclear without a specific definition, but should be understood according to the ordinary meaning in the art. When a trade name appears herein, it is intended to refer to its corresponding product or active ingredient thereof.

[0212] Herein represents a point of attachment.

[0213] Herein, a bond depicted by a solid line and a dashed line represents a single bond or a double bond.

[0214] Unless otherwise indicated, a wedge and a dashed wedge represents the absolute configuration of a stereocenter. A wavy line represents an unspecified configuration, i.e., can be either the Z or E configuration, or both configurations.

[0215] The term "tautomers" refers to isomers of functional groups that result from the rapid movement of an atom in a molecule between two positions. The compounds of the present disclosure can exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible forms. Tautomers generally exist in equilibrium and attempts to isolate a single tautomer usually result in a mixture whose physical and chemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; in phenols, the enol form predominates. The present disclosure includes all tautomeric forms of the compounds. The term "stereoisomers" refers to isomers having the same molecular formula but different physical properties, such as different melting points, boiling points, and / or different reactivities in chemical reactions. Stereoisomers include enantiomers (mirror images of the other that are not superimposable), and diastereomers (isomers that are not mirror images of each other).

[0216] The compounds of the present disclosure can have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms, or asymmetric double bonds, and thus the compounds of the present disclosure can exist in particular geometric or stereoisomeric forms. The particular geometric or stereoisomeric forms can be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as racemic mixtures or other mixtures thereof, such as those that contain one enantiomeric or diastereomeric excess, all of which are within the scope of the compounds of the present disclosure. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms can be present in a substituent group, and all such isomers and mixtures thereof are included within the definition of the compounds of the present disclosure. The compounds of the present disclosure containing an asymmetric atom can be isolated in optically active form or as racemic mixtures, and the optically active forms can be obtained by separation from the racemic mixtures or by using chiral starting materials or chiral reagents in the synthetic sequence.

[0217] The term "substituted" means that any one or more hydrogen atoms on the particular atom is replaced with a substituent group, provided that the valence of the particular atom is not normally exceeded, and that the substituted compound is stable. When the substituent is oxo (i.e., =0), it means that two hydrogen atoms are replaced by the substituent.

[0218] The term "optionally" or "optional" means that the subsequently described event or circumstance can or can not occur, and this description includes instances where the event or circumstance occurs and instances where it does not. For example, an ethyl group "optionally" substituted with a halogen means that the ethyl group can be unsubstituted (CH2CH3), mono-substituted (CH2CH2F, CH2CH2C1, etc.), poly-substituted (CHFCH2F, CH2CHF2, CHFCH2C1, CH2CHC12, etc.), or fully substituted (CF2CF3, CF2CC13, CC12CC13, etc.). One skilled in the art will appreciate that for any given group containing one or more substituents, no substitution or substitution pattern is introduced that is not chemically possible.

[0219] When any variable (e.g., R a , R b ) occurs more than one time in a compound, its definition in each occurrence is independent of its definition at every other occurrence. For example, if a group is substituted with 2 R b groups, then each R b group is independently selected.

[0220] C m -C nIt means having an integer number of carbon atoms in the range mn. For example, "C1-C 10 ” means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

[0221] The term "alkyl" refers to a group of the formula C n H 2n+1 The term "C1-C 10 The term "alkyl" is understood to mean a straight-chain or branched saturated hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Specific examples of the alkyl radical include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2- dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; the term "C1-C6 alkyl" can be understood to mean an alkyl group having 1 to 6 carbon atoms, and specific examples include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C3 alkyl" can be understood to mean a straight-chain or branched saturated alkyl group having 1 to 3 carbon atoms. The "C1-C 10 The term "alkyl" may include "C1-C6 alkyl" or "C1-C3 alkyl" and the like, and the "C1-C6 alkyl" may further include "C1-C3 alkyl".

[0222] The term "alkoxy" refers to a group generated by the loss of a hydrogen atom from a hydroxyl group of a straight-chain or branched alcohol, and can be understood as "alkyloxy" or "alkyl-O-". The term "C1-C 10 "Alkoxy" can be understood as "C1-C 10 Alkyloxy" or "C1-C 10 Alkyl-O-"; the term "C1-C6 alkoxy" can be understood as "C1-C6 alkyloxy" or "C1-C6 alkyl-O-". The "C1-C 10 The term "alkoxy" may include "C1-C6 alkoxy" and "C1-C3 alkoxy", and the "C1-C6 alkoxy" may further include "C1-C3 alkoxy".

[0223] The term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one double bond. 10 "Alkenyl" is understood to mean a linear or branched unsaturated hydrocarbon radical containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, "C2-C 10 "Alkenyl" is preferably "C2-C6 alkenyl", further preferably "C2-C4 alkenyl", and further preferably C2 or C3 alkenyl. It will be understood that when the alkenyl contains more than one double bond, the double bonds may be separated from each other or conjugated. Specific examples of the alkenyl include, but are not limited to, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl or (Z)-1-methylprop-1-enyl, etc.

[0224] The term "cycloalkyl" refers to a fully saturated carbon ring that exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 10-membered ring. The term "C3-C 10 The term "cycloalkyl" is understood to mean a saturated monocyclic, bicyclic, spirocyclic or bridged ring having 3 to 10 carbon atoms. Specific examples of the cycloalkyl group include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, and the like. The term "C3-C 10 The term "cycloalkyl" may include "C3-C6 cycloalkyl". The term "C3-C6 cycloalkyl" may be understood to mean a saturated monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms, and specific examples include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. The term "C3-C4 cycloalkyl" may be understood to mean a saturated monocyclic or bicyclic hydrocarbon ring having 3 to 4 carbon atoms, and specific examples include but are not limited to cyclopropyl or cyclobutyl.

[0225] The term "phenylene" refers to a divalent group derived from "phenyl".

[0226] The term "heterocyclyl" refers to a monocyclic, bicyclic, spirocyclic, or bridged cyclic radical that is fully saturated or partially saturated (i.e., is not a heteroaromatic radical that is aromatic overall), having from one to five heteroatoms or heteroatom groups (i.e., groups of atoms that contain heteroatoms) in its ring atom count, including, but not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (B), -S(=0)2-, -S(=0)-, -P(=0)2-, -P(=0)-, -NH-, -S(=0)(=NH)-, -C(=0)NH-, or -NHC(=0)NH-, and the like, in its ring atom count.

[0227] The term "4-12 membered heterocyclyl" refers to a heterocyclyl group having a ring atom number of 4, 5, 6, 7, 8, 9, 10, 11, or 12, and having 1-5 ring atoms independently selected from the above-mentioned heteroatoms or heteroatom groups. The "4-12 membered heterocyclyl" includes "4-10 membered heterocyclyl", "4-7 membered heterocyclyl", "4-6 membered heterocyclyl", and the like, wherein specific examples of 4-membered heterocyclyl include, but are not limited to, azetidinyl or oxetanyl; specific examples of 5-membered heterocyclyl include, but are not limited to, tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl, or 2,5-dihydro-1H-pyrrolyl; specific examples of 6-membered heterocyclyl include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl, or 4H-[1,3,4]thiadiazinyl; and specific examples of 7-membered heterocyclyl include, but are not limited to, diazepanyl. The heterocyclyl group can also be a bicyclic group, wherein specific examples of 5,5 membered bicyclic group include, but are not limited to, hexahydrocyclopenta[c]pyrrol-2(1H)-yl; specific examples of 5,6 membered bicyclic group include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl, or 5,6,7,8-tetrahydroimidazolo[1,5-a]pyrazinyl. Optionally, the heterocyclyl group can be a benzo-fused ring group of the above-mentioned 4-7 membered heterocyclyl, and specific examples include, but are not limited to, dihydroisoquinolinyl and the like. The "4-10 membered heterocyclyl" can include "5-10 membered heterocyclyl", "4-7 membered heterocyclyl", "5-6 membered heterocyclyl", "6-8 membered heterocyclyl", "4-10 membered heterocycloalkyl", "5-10 membered heterocycloalkyl", "4-7 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", "6-8 membered heterocycloalkyl", and the like, and the "4-7 membered heterocyclyl" can further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocycloalkyl", "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", and the like. The above-mentioned heterocyclyl or heterocycloalkyl group can have 1, 2, or 3 heteroatoms independently selected from N, O, and S, or can have 1 or 2 heteroatoms independently selected from N and O. Although some bicyclic heterocyclyl moieties in the present disclosure partially contain one benzene ring or one heteroaromatic ring, the heterocyclyl group as a whole is still non-aromatic.

[0228] The term "heterocycloalkyl" refers to a fully saturated cyclic group, existing in the form of monocyclic, annelated, bridged or spirocyclic, etc., having 1-5 heteroatoms or heteroatom groups (i.e., an atom group containing a heteroatom) in its ring atoms, said "heteroatoms or heteroatom groups" including, but not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O)2-, -S(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH-, or -NHC(=O)NH-, etc. The term "4-10 membered heterocycloalkyl" refers to a heterocycloalkyl group having 4, 5, 6, 7, 8, 9, or 10 ring atoms, and having 1-5 ring atoms independently selected from the above-mentioned heteroatoms or heteroatom groups. "4-10 membered heterocycloalkyl" includes "4-7 membered heterocycloalkyl", wherein specific examples of 4-membered heterocycloalkyl groups include, but are not limited to, azetidinyl (azetidinyl), oxetanyl (oxetanyl), or thietanyl (thietanyl); specific examples of 5-membered heterocycloalkyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl, or tetrahydropyrazolyl; specific examples of 6-membered heterocycloalkyl groups include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1,4-thioxanyl, 1,4-dioxanyl, thiomorpholinyl, 1,3-dithianyl, or 1,4-dithianyl; specific examples of 7-membered heterocycloalkyl groups include, but are not limited to, azepanyl, oxepanyl, or thiepanyl.

[0229] The term "heterocyclyl" refers to a divalent group derived from a "heterocycloalkyl".

[0230] The term "heteroaryl" refers to a monocyclic or fused polycyclic aromatic system containing at least one (e.g., 1, 2, or 3) ring atom selected from N, O, S, with the remaining ring atoms being C, typically having 5 to 14 members, 5 to 12 members, 5 to 10 members, 5 to 8 members, 5 to 7 members, or 5 to 6 members in the ring. Preferred heteroaryls have a single 4 to 8 member ring, especially 5 to 6 member rings, or multiple fused rings containing 5 to 14, especially 5 to 10 ring atoms. Non-limiting examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothiophenyl, indolyl, isoindolyl, and the like.

[0231] The term "heteroarylene" refers to a divalent group derived from a "heteroaryl".

[0232] The term "halo" or "halogen" refers to fluoro, chloro, bromo, or iodo.

[0233] The term "hydroxy" refers to an -OH group.

[0234] The term "cyano" refers to a -CN group.

[0235] The term "amino" refers to a -NH2 group.

[0236] The term "therapeutically effective amount" means an amount of a compound of the present disclosure that (i) treats a particular disease, condition, or disorder, or (ii) attenuates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder. The amount of a compound of the present disclosure that will constitute a "therapeutically effective amount" will vary depending on the compound, the disease state and its severity, the manner of administration, and the age of the mammal to be treated, but can be determined routinely by the skilled practitioner by a consideration of the factors relevant to the choice of an appropriate dose of a therapeutic compound.

[0237] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0238] The term "pharmaceutically acceptable salt" refers to a salt of an acid or base which is pharmaceutically acceptable, including salts of inorganic acids or organic acids, and salts of inorganic bases or organic bases.

[0239] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or salts thereof with a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of a compound of the present disclosure to an organism.

[0240] The term "pharmaceutically acceptable excipient" refers to an excipient that is not biologically or otherwise undesirable, i.e., the excipient can be administered to an organism without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. The presence of an excipient in a composition does not imply that the excipient is intended to be part of the compound of the present disclosure.

[0241] The words "comprise" or "comprising" and variations thereof such as "comprises" or "comprising", when used in this document, can be understood to encompass the terms "consisting of" or "consisting of".

[0242] The present disclosure also includes isotopically-labeled compounds of the present disclosure which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be present in compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, iodine, and chlorine, such as 2 H, 3 H,11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 123 I, 125 I and 36 Cl, and the like.

[0243] Certain isotopically-labeled compounds of the present disclosure (e.g., with 3 H and 14 C) can be used in compound and / or substrate tissue distribution analysis. Tritiated (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred for their ease of preparation and detectability. Positron emitting isotopes such as 15 O, 13 N, 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically-labeled compounds of the present disclosure can generally be prepared by substituting a readily available isotopically-labeled reagent for a non-isotopically labeled reagent in a procedure similar to those disclosed in the schemes and / or examples below.

[0244] The pharmaceutical compositions of the present disclosure can be prepared by combining a compound of the present disclosure with suitable pharmaceutically acceptable excipients, and can be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, pills, capsules, powders, granules, ointments, creams, suspensions, suppositories, injections, inhalants, gels, microspheres, aerosols and the like.

[0245] Typical routes of administering a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, include, but are not limited to, oral, rectal, topical, inhalant, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

[0246] The pharmaceutical compositions of the present disclosure can be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, emulsifying, lyophilizing processes, and the like.

[0247] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by combining the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, dragees, sugar-coated tablets, capsules, liquids, gels, slurries, suspensions, and the like for oral administration to a patient.

[0248] Solid oral compositions can be prepared by conventional mixing or encapsulating processes. For example, the active compounds can be mixed with a solid excipient, optionally ground, and then filled into a capsule, if desired with the addition of additional excipients. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, or flavoring agents.

[0249] The pharmaceutical compositions can also be in a form suitable for parenteral administration, such as an aqueous or non-aqueous sterile injection solution or suspension, or a lyophilized product.

[0250] In all methods of administration of the compounds of general formula (I) described herein, the daily dose can range from 0.01 mg / kg to 1000 mg / kg of body weight, in single or divided doses.

[0251] The compounds of the present disclosure can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by a combination of the other chemical synthetic methods well known to those skilled in the art, and equivalent alternatives well known to those skilled in the art, preferred embodiments including but not limited to the examples of the present disclosure.

[0252] The chemical reactions of the specific embodiments of the present disclosure are performed in solvents appropriate to the reagents and materials employed and suitable for the chemical changes being effected. In order to obtain the compounds of the present disclosure, it can be necessary to modify the synthetic procedures described in the embodiments or to choose alternative synthetic routes. BRIEF DESCRIPTION OF DRAWINGS

[0253] Figure 1 is a bar graph of the in vitro thrombolytic activity of compound 001.

[0254] Figure 2 is a bar graph of the antioxidant activity of the compounds in each group, wherein (A) and (B) are bar graphs of the antioxidant activity of the compounds in each group at a concentration of 150 μM, and (C) is a bar graph of the antioxidant activity of the compounds in each group at a concentration of 250 μM; wherein "*" represents a statistical difference compared to the control molecule, * p < 0.05, ** p < 0.01, *** p < 0.001, ****p<0.0001.

[0255] Figure 3 is a bar graph of the infarct volume of the efficacy evaluation test of Example 6 control molecules with different administration methods.

[0256] Figure 4 is a graph of the bleeding risk evaluation test of Example 6 control molecules with different administration methods, wherein (A) is a graph of the bleeding condition of the model control (Model) group of rats, (B) is a graph of the bleeding condition of the control molecule bolus+infusion group of rats; (C) is a graph of the bleeding condition of the control molecule bolus group of rats.

[0257] Figure 5 is a bar graph of the infarct volume of the efficacy evaluation test of Example 7 compound 023.

[0258] Figure 6 is a graph of the bleeding risk evaluation test of Example 7 compound 023, wherein (A) is a graph of the bleeding condition of the model control (Model) group of rats, (B) is a graph of the bleeding condition of the control molecule bolus+infusion group of rats; (C) is a graph of the bleeding condition of the compound 023 bolus group of rats.

[0259] Figure 7 is a bar graph of the percentage of infarct volume of the efficacy test of Example 8 tMCAO.

[0260] Figure 8 is a graph of the TTC staining results of Example 8 tMCAO efficacy test, wherein (A) is a graph of the TTC staining results of the vehicle control (Vehicle) group of rats, (B) is a graph of the TTC staining results of the control molecule bolus group of rats; (C) is a graph of the TTC staining results of the compound 057 bolus group of rats.

[0261] Figure 9 is a bar graph of the bleeding score of the efficacy test of Example 8 tMCAO.

[0262] Figure 10 is a graph of the bleeding condition of the rats of the efficacy test of Example 8 tMCAO, wherein (A) is a graph of the bleeding condition of the vehicle control (Vehicle) group of rats, (B) is a graph of the bleeding condition of the control molecule bolus group of rats; (C) is a graph of the bleeding condition of the compound 057 bolus group of rats. DETAILED DESCRIPTION

[0263] The application is described in detail below by way of examples, but it is not meant to be any unfavorable limitation on the disclosure. The disclosure has been described in detail herein, and specific embodiments thereof have also been disclosed, it will be obvious to those skilled in the art that various changes and improvements can be made to the specific embodiments of the disclosure without departing from the spirit and scope of the disclosure. All reagents used in the disclosure are commercially available and can be used without further purification.

[0264] Unless otherwise stated, the ratio indicated for mixed solvents is the ratio by volume of the mixture.

[0265] The compounds were named by hand or by software, and commercially available compounds were named using the supplier's catalogue name. The compounds were named by hand or by software, and commercially available compounds were named using the supplier's catalogue name.

[0266] The structure of the compounds was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The unit of NMR shift is 10 -6 (ppm). The solvent for NMR determination was deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard was tetramethylsilane (TMS); "IC 50 "IC

[0267] The eluent hereinafter can be formed by two or more solvents in a mixed eluent, and the ratio is the ratio by volume of each solvent.

[0268] Example 1: Synthesis of 2-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoyl)oxy)acetic acid (Compound 001)

[0269] Step 1: Synthesis of methyl (1,3-dioxoisoindolin-2-yl)-2-hydroxyacetate (Intermediate 1-3)

[0270] Hydroxyacetic acid (200 mg, 2.63 mmol), N-bromomethylphthalimide (694 mg, 2.89 mmol) and N,N-diisopropylethylamine (680 mg, 5.26 mmol) were dissolved in acetone (6 mL), and the reaction solution was stirred at room temperature for 3 hours. The reaction solution was concentrated, and the residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 2-1 / 1) to obtain the title compound 1-3 (250 mg).

[0271] 1 H NMR (400 MHz, CDCl3) δ 4.19 (s, 2H), 5.84 (s, 2H), 7.80-7.82 (m, 2H), 7.94-7.94 (m, 2H).

[0272] Step two: synthesis of 2-((l,3-dioxoisoindolin-2-yl)methoxy)-2-oxoethyl-(S)-2,5- bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-l-yl)-3,5-dihydroxy-2-methyl-7-oxo- 3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoate (Intermediate 1-4)

[0273] Compound 1-5 (110 mg, 127 μmol, prepared by microbial fermentation according to patent JP2004-224738A example 2, which is SMTP-7L, i.e. compound 1-5), Intermediate 1-3 (31 mg, 132 μmol) and O-(7-azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (57 mg, 150 μmol) were dissolved in N,N-dimethylformamide (2.2 mL), and N,N-diisopropylethylamine (32 mg, 254 μmol) was added. The reaction was stirred at room temperature for 4.5 hours. LCMS showed the reaction was completed. The reaction was added dropwise into water (12 mL), stirred for 20 minutes, filtered, and the filter cake was collected to give the crude title compound 1-4 (110 mg), which was used directly in the next step.

[0274] MS (ESI): m / z = 1086.5 [M+H] + .

[0275] Step three: synthesis of 2-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-l-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoyl)oxy)acetic acid (Compound 001)

[0276] Compound 1-4 (110 mg, 101 μmol) was dissolved in methanol (2.5 mL), and hydrazine hydrate (17.8 mg, 303 μmol, purity: 85%) was added. The reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The reaction was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent] and lyophilized to give the title compound 001 (10 mg).

[0277] MS (ESI): m / z = 927.6 [M+H] + .

[0278] 1H NMR (400 MHz, DMSO-d6) δ 1.20 - 1.12 (m, 6 H), 1.48 - 1.64 (m, 24 H), 1.86 - 2.02 (m, 10 H), 2.06 - 2.15 (m, 4 H), 2.42 - 2.48 (m, 2 H), 2.81 (dt, J = 17.2, 5.6 Hz, 2 H), 3.47 (t, J = 7.2 Hz, 2 H), 3.70 - 3.77 (m, 2 H), 4.13 - 4.23 (m, 3 H), 4.29 - 4.40 (m, 3 H), 4.92 (dd, J = 10.8, 5.2 Hz, 1 H), 5.01 - 5.06 (m, 2 H), 5.08 - 5.20 (m, 4 H), 6.62 (s, 1 H), 6.67 (s, 1 H), 9.83 (br s, 1 H), 9.88 (br s, 1 H).

[0279] Example 2: Synthesis of (((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido)methyl)phosphonic acid (Compound 002)

[0280] Compound 2-1 (70 mg, 630.42 μmol) was dissolved in dichloromethane (3 mL), diisopropylethylamine (244.43 mg, 1.89 mmol) and trimethylsilyl chloride (136.98 mg, 1.26 mmol) were added. The reaction was stirred at 25 °C for 3 hours, then compound 1-5 (109.58 mg, 126.08 μmol), 1-hydroxy-7-azabenzotriazole (34.3 mg, 252.17 μmol) and N,N'-diisopropylcarbodiimide (79.56 mg, 630.42 μmol) were added. The reaction was then stirred at 25 °C for 16 hours. LCMS showed the reaction was completed. The reaction was added with dichloromethane (20 mL) and washed with 1 N aqueous HC1 (5 mL). The organic phase was dried and concentrated. The crude product was purified by prep-HPLC [column: Phenomenex Gemini NX 150 x 30 mm, 5 μm; mobile phase: [water (0.05% ammonia water-0.001% ammonium bicarbonate)-acetonitrile]; gradient: 35%-55%] to give the title compound 002 (19.1 mg).

[0281] MS (ESI): m / z = 962.5 [M+H] + .

[0282] 1H NMR (400 MHz, DMSO-d6) δ 7.82 (br s, 1H), 6.73 - 6.65 (m, 2H), 5.15 - 5.02 (m, 4H), 4.94 - 4.85 (m, 1H), 4.56 - 4.46 (m, 1H), 4.22 - 4.10 (m, 3H), 3.80 - 3.68 (m, 2H), 3.56 - 3.41 (m, 4H), 3.09 (dt, J = 3.7, 13.1 Hz, 2H), 2.86 - 2.76 (m, 2H), 2.48 - 2.38 (m, 2H), 2.20 - 1.72 (m, 14H), 1.67 - 1.29 (m, 24H), 1.20 - 1.10 (m, 6H).

[0283] Example 3: Synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido) propanoic acid (Compound 003)

[0284] Step one: Synthesis of 2,5-dioxopyrrolidin-1-yl (S)-2,5-bis((2R,3S)-2-(((E)-4,8-dimethylnona- 3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoate

[0285] Step one: Synthesis of 2,5-dioxopyrrolidin-1-yl (S)-2,5-bis((2R,3S)-2-(((E)-4,8-dimethylnona- 3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoate

[0286] MS (ESI): m / z = 966.5 [M+H] + .

[0287] Step two: Synthesis of methyl 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido) propanoate

[0288] Compound 3-1 (300 mg, 310.51 μmol), 3-amino propionic acid ethyl ester hydrochloride (130.02 mg, 931.52 μmol), 4-dimethylaminopyridine (37.93 mg, 310.51 μmol) and N,N-diisopropylethylamine (200.65 mg, 1.55 mmol) were added into N,N-dimethylformamide (5 mL). The reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The reaction was diluted with 30 mL water, extracted with ethyl acetate (50 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure to give the crude compound 3-2 (320 mg).

[0289] MS (ESI): m / z = 954.3 [M+H] + .

[0290] Step three: synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido)propanoic acid

[0291] Compound 3-2 (80 mg, 83.84 μmol), lithium hydroxide monohydrate (10.55 mg, 251.52 μmol) were added into tetrahydrofuran (1 mL) and water (0.5 mL). The reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The reaction was concentrated, the residue was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 80%-90%) as eluent] and lyophilized to give the title compound 003 (30 mg).

[0292] MS (ESI): m / z = 940.3 [M+H] + .

[0293] 1H NMR (400 MHz, DMSO-d6) δ 8.15 (s, 1H), 6.65 (s, 1H), 6.63 (s, 1H), 5.18 - 4.97 (m, 6H), 4.75 - 4.67 (m, 1H), 4.40 (d, J = 17.1 Hz, 1H), 4.21 - 4.12 (m, 3H), 3.78 - 3.69 (m, 2H), 3.49 - 3.44 (m, 2H), 3.25 - 3.15 (m, 2H), 2.88 - 2.76 (m, 2H), 2.51 - 2.40 (m, 2H), 2.31 - 2.22 (m, 2H), 2.18 - 1.84 (m, 14H), 1.68 - 1.48 (m, 24H), 1.18 (s, 3H), 1.15 (s, 3H).

[0294] Example 4: Synthesis of ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoyl)- L-lysine (Compound 004)

[0295] Step one: Synthesis of N6-(((9H-fluoren-9-yl)methoxy)carbonyl)-N2-((S)-2,5-bis((2R,3S)- 2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9- tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoyl)-L-lysine

[0296] (2S)-6-(9H-fluoren-9-ylmethoxycarbonylamino)-2-aminohexanoic acid (114.40 mg, 310.51 μmol), compound 3-1 (100 mg, 103.50 μmol), 4-dimethylaminopyridine (12.64 mg, 103.50 μmol) and N,N-diisopropylethylamine (53.51 mg, 414.01 μmol) were added into N,N-dimethylformamide (3 mL), the reaction was stirred at room temperature for 2 hours. LCMS showed that the reaction was completed. 30 mL of water was added to dilute the reaction, extracted with ethyl acetate (50 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give the crude title compound 4-1 (110 mg).

[0297] MS (ESI): m / z = 1219.5 [M+H] + .

[0298] Step two: synthesis of ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoyl)- L-lysine

[0299] Compound 4-1 (200 mg, 164.00 μmol) and triethylamine (99.57 mg, 984.01 μmol) were added into methanol (5 mL), the reaction was stirred at 60 °C for 18 hours. LCMS showed the reaction was completed. The reaction was filtered, purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 85%-90%) as eluent], and then lyophilized to obtain the title compound 004 (10 mg).

[0300] MS (ESI): m / z = 997.7 [M+H] + .

[0301] 1 H NMR (400 MHz, DMSO-d6) δ 7.47 (s, 1H), 6.67 (s, 1H), 6.63 (s, 1H), 5.23 - 4.95 (m, 6H), 4.80 - 4.71 (m, 1H), 4.31 (d, J = 17.5 Hz, 1H), 4.19 - 4.11 (m, 3H), 3.76 - 3.71 (m, 2H), 3.49 - 3.44 (m, 2H), 2.86 - 2.77 (m, 4H), 2.63 - 2.58 (m, 2H), 2.46 - 2.41 (m, 2H), 2.21 - 1.82 (m, 16H), 1.68 - 1.43 (m, 26H), 1.16 (s, 3H), 1.14 (s, 3H), 0.89 - 0.81 (m, 2H).

[0302] Example 5: synthesis of (2R,2'R,3S,3'S)-8,8'-((S)-5-hydroxypentane-1,4-diyl)bis(2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-3,4,8,9-tetrahydropyrano[2,3-E]isoindol- 7(2H)-ketone) (compound 005)

[0303] Step 1: Synthesis of (S)-methyl 2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoate

[0304] Compound 1-5 (500 mg, 575.31 μmol), sodium bicarbonate (144.98 mg, 1.73 mmol), iodomethane (245.08 mg, 1.73 mmol) were added into N,N-dimethylformamide (5 mL). The reaction was stirred at room temperature for 18 hours. LCMS showed the reaction was completed. The reaction was cooled and poured into water (20 mL), extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure to give the crude title compound 5-1 (510 mg), which was used directly in the next step.

[0305] MS (ESI): m / z = 883.5 [M+H] + .

[0306] Step 2: Synthesis of (2R,2'R,3S,3'S)-8,8'-((S)-5-hydroxypentane-1,4-diyl)bis(2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-3,4,8,9-tetrahydropyrano[2,3- E]isoindol-7(2H)-ketone)

[0307] 5-1 (50 mg, 56.62 μmol) was added into tetrahydrofuran (2 mL). Lithium aluminum hydride (4.30 mg, 113.24 μmol) was added under ice water bath. The reaction was stirred at -20 °C for 0.5 hours. LCMS showed the reaction was completed. The reaction was quenched by adding Na2SO4·10H2O, filtered, the filtrate was concentrated, the residue was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent] and lyophilized to give the title compound 005 (10 mg).

[0308] MS (ESI): m / z = 855.3 [M+H] + .

[0309] 1H NMR (400 MHz, DMSO-d6) δ 9.72 (s, 1H), 9.71 (s, 1H), 6.63 (s, 1H), 6.61 (s, 1H), 5.16 - 5.07 (m, 4H), 5.05 (s, 2H), 4.75 (t, J = 5.5 Hz, 1H), 4.22 - 4.04 (m, 4H), 3.76 - 3.68 (m, 2H), 3.55 - 3.38 (m, 4H), 2.87 - 2.75 (m, 2H), 2.47 - 2.40 (m, 2H), 2.16 - 1.82 (m, 14H), 1.69 - 1.37 (m, 24H), 1.18 - 1.13 (m, 6H).

[0310] Example 6: Synthesis of 2-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentyl)oxy)acetic acid (Compound 006)

[0311] Step one: Synthesis of (S)-methyl 2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3-hydroxy-5-(methoxymethoxy)-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)- yl)pentanoate

[0312] Compound 5-1 (1.5 g, 1.70 mmol) and N,N-diisopropyl ethylamine (878.07 mg, 6.79 mmol) were added into dichloromethane (15 mL). Bromomethyl methyl ether (1.06 g, 8.49 mmol) was added under ice water bath. The reaction was stirred at room temperature for 0.5 h. LCMS showed the reaction was completed. 40 mL water was added for dilution, extracted with ethyl acetate (50 mL*3), washed with saturated brine (50 mL*4), dried and concentrated to get the crude title compound 6-1 (1.7 g). Used directly for next step.

[0313] MS (ESI): m / z = 971.3 [M+H] + .

[0314] Step two: Synthesis of (2R,2'R,3S,3'S)-8,8'-((S)-5-hydroxypentane-1,4-diyl)bis(2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3-hydroxy-5-(methoxymethoxy)-2-methyl-3,4,8,9- tetrahydropyrano[2,3-E]isoindol-7(2H)-ketone)

[0315] Compound 6-1 (1 g, 1.03 mmol) was added to tetrahydrofuran (15 mL). Lithium aluminum hydride (234.45 mg, 6.18 mmol) was added under ice water bath. The reaction was stirred at room temperature for 0.5 h. LCMS showed the reaction was completed. The reaction was quenched by adding Na2SO4·10H2O, and the reaction was poured into water (20 mL) after cooling, extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica, dichloromethane / methanol = 10 / 1) to give the title compound compound 6-2 (300 mg).

[0316] MS (ESI): m / z = 943.3 [M+H] + .

[0317] Step three: synthesis of ethyl 2-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3-hydroxy-5-(methoxymethoxy)-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentyl)oxy)acetate Compound 6-2 (50 mg, 53.01 µmol) was added to dichloromethane (1 mL), sodium hydride (12.72 mg, 318.06 µmol, 60% purity) was added under ice water bath. The reaction was stirred at room temperature for 1 h. Ethyl bromoacetate (26.56 mg, 159.03 µmol) was added under ice water bath. The reaction was stirred at room temperature for 6 h. LCMS showed the reaction was completed. The reaction was quenched by adding water (5 mL), extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give the crude title compound 6-3 (50 mg).

[0318] MS (ESI): m / z = 1029.3 [M+H] + .

[0319] Step four: synthesis of ethyl 2-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentyl)oxy)acetate

[0320] Compound 6-3 (30 mg, 29.55 μmol), 4-methylbenzenesulfonic acid pyridine (22.28 mg, 88.65 μmol) were added into tert-butanol (2 mL). The reaction was stirred at 80 °C for 18 h. The reaction was quenched by adding water (10 mL), extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give the crude title compound 6-4 (55 mg)

[0321] MS (ESI): m / z = 941.3 [M+H] + .

[0322] Step five: synthesis of 2-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentyl)oxy)acetic acid

[0323] Compound 6-4 (50 mg, 53.93 μmol), lithium hydroxide monohydrate (6.79 mg, 161.78 μmol) were added into tetrahydrofuran (1 mL) and water (1 mL), the reaction was stirred at room temperature for 18 h. The reaction was quenched by adding 2 mL 1M aqueous HC1 to adjust pH = 5, extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated, the residue was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent], lyophilized to give the title compound 006 (1 mg).

[0324] MS (ESI): m / z = 913.3 [M+H] + .

[0325] 1H NMR (400 MHz, DMSO-d6) δ 9.85 (s, 1H), 9.75 (s, 1H), 6.68 (s, 1H), 6.62 (s, 1H), 5.20 - 4.94 (m, 6H), 4.24 - 4.05 (m, 4H), 3.76 - 3.59 (m, 2H), 3.48 - 3.39 (m, 2H), 3.33 - 3.24 (m, 2H), 2.86 - 2.76 (m, 2H), 2.50 - 2.37 (m, 2H), 2.20 - 1.74 (m, 14H), 1.68 - 1.37 (m, 24H), 1.28 - 1.07 (m, 6H).

[0326] Example 7: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanal oxime (Compound 007)

[0327] Step one: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)-N- methoxy-N-methylpentanamide

[0328] Compound 3-1 (200 mg, 207 μmol), N, O-dimethylhydroxylamine hydrochloride (63.19 mg, 647.88 μmol), 4-dimethylaminopyridine (26.38 mg, 215.96 μmol) and N, N- diisopropylethylamine (139.55 mg, 1.08 mmol) were added into N, N-dimethylformamide (3 mL). The reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The reaction was poured into water (20 mL), extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, the residue was purified by column chromatography (silica, dichloromethane / methanol = 10 / 1) to give the title compound 7-1 (50 mg).

[0329] MS (ESI): m / z = 912.3 [M+H] + .

[0330] Step two: synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanal

[0331] Step two: synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanal

[0332] MS (ESI): m / z = 853.3 [M+H] + .

[0333] Step three: synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanal oxime

[0334] Step two: synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanal

[0335] MS (ESI): m / z = 868.3 [M+H] + .

[0336] 1H NMR (400 MHz, DMSO-d6) δ 10.81 (s, 1H), 9.78 (s, 1H), 9.74 (s, 1H), 7.42 (d, J = 4.9 Hz, 1H), 6.64 (s, 1H), 6.62 (s, 1H), 5.38 - 4.99 (m, 6H), 4.87 - 4.77 (m, 1H), 4.29 - 4.10 (m, 4H), 3.77 - 3.69 (m, 2H), 3.49 - 3.44 (m, 2H), 2.87 - 2.63 (m, 2H), 2.50 - 2.40 (m, 2H), 2.20 - 1.83 (m, 14H), 1.68 - 1.41 (m, 24H), 1.18 - 1.14 (m, 6H).

[0337] Example 8: Synthesis of (2R,2'R,3S,3'S)-8,8'-((S)-5-hydroxy-5-methylhexane-1,4-diyl) bis(2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-3,4,8,9- tetrahydropyrano[2,3-E]isoindol-7(2H)-one) (Compound 008)

[0338] Compound 5-1 (50 mg, 56.62 μmol) was dissolved in dry tetrahydrofuran (3 mL), and a solution of methyl magnesium bromide (1 M in THF, 1.2 mL, 1.2 mmol) was added dropwise to the reaction mixture while maintaining the temperature of the reaction mixture at no more than -60 °C. The reaction mixture was then warmed to 0 °C and stirred for 2 hours. The reaction mixture was added dropwise to an aqueous solution of hydrogen chloride (0.5 M, 10 mL), stirred for 2 minutes, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 65-85%) as eluent] and the title compound 008 (10 mg) was obtained after lyophilization.

[0339] 1H NMR (400 MHz, DMSO-d6) δ 9.76 (br s, 2H), 6.65 (s, 1H), 6.61 (s, 1H), 5.07-5.20 (m, 4H), 4.99-5.07 (m, 2H), 4.58 (br s, 1H), 4.43 (d, J = 17.8 Hz, 1H), 4.19-3.92 (m, 4H), 3.70-3.77 (m, 2H), 3.55-3.45 (m, 2H), 2.80 (dt, J = 17.5, 5.4 Hz, 2H), 2.42-2.48 (m, 2H), 2.03-2.18 (m, 4H), 1.69-2.02 (m, 10H), 1.48-1.64 (m, 24H), 1.19 (s, 3H), 1.15 (s, 3H), 1.14 (s, 3H), 0.92 (s, 3H).

[0340] Example 9: Synthesis of (2S)-2,5-bis((2R)-3-amino-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid (Compound 009)

[0341] Step one: Synthesis of (2R,2'R)-((S)-5-methoxy-5-oxopentane-1,4-diyl) bis(2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3-hydroxy-2-methyl-7-oxo-2,3,4,7,8,9-hexahydropyrano[2,3-E]isoindol-8,5-diyl) diacetate

[0342] Compound 5-1 (500.0 mg, 566.18 μmol) and N,N-diisopropylethylamine (219.5 mg, 1.70 mmol) were dissolved in dichloromethane (8.0 mL). Acetyl chloride (133.3 mg, 1.70 mmol) was added dropwise at 0 °C, and the dropwise addition was completed in 5 min. The reaction solution was stirred at room temperature for 2 h. The reaction was monitored by LCMS, quenched with saturated sodium bicarbonate solution, extracted with dichloromethane (20.0 mL*3), and the combined organic phase was washed once with saturated brine (20.0 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 1 / 4) to give the title compound 9-1 (320.0 mg).

[0343] MS (ESI): m / z = 966.7 [M+H] + .

[0344] Step two: synthesis of (S)-methyl 5-((R)-5-acetyloxy-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 2-methyl-3,7-dioxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)-2-((R)-5- acetyloxy-2-methyl-2-((E)-8-nonadien-1-yl)-3,7-dioxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)valerate

[0345] Compound 9-1 (320.0 mg, 330.86 μmol) was dissolved in dichloromethane (6.0 mL). At 0 °C, Dess-Martin reagent (421.0 mg, 992.57 μmol) was added, and the reaction was stirred at room temperature for 0.5 h. LCMS showed the reaction was complete. The reaction was quenched with saturated aqueous sodium thiosulfate solution, extracted with dichloromethane (20.0 mL*3), and the combined organic phase was washed with saturated brine (20.0 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 1 / 1) to give intermediate 9-2 (250.0 mg).

[0346] Step three: synthesis of (S)-methyl 5-((R,E)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5- hydroxy-3-(hydroxyimino)-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)- 2-((R,Z)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5-hydroxy-3-(hydroxyimino)-2-methyl-7- oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)valerate

[0347] Compound 9-2 (250.0 mg, 263.39 μmol) and hydroxylamine hydrochloride (54.1 mg, 778.69 μmol) were dissolved in ethanol (6.0 mL). At 25 °C, sodium acetate (63.8 mg, 778.69 μmol) was added, and the reaction was stirred at room temperature for 0.5 h. LCMS showed the reaction was complete. The ethanol was removed by concentration under reduced pressure, and the residue was purified by prep-HPLC [column: C-18 (spherical 40-60 μm 100A 40 g); mobile phase: [water (0.1% formic acid) - acetonitrile]; gradient: 0% - 75%] to give intermediate 9-3 (130.0 mg).

[0348] Step four: Synthesis of methyl (2S)-2,5-bis((2R)-3-amino-2-((E)-4,8-dimethyl- nona-3,7-dien-1-yl)-5-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanoate

[0349] Compound 9-3 (70.0 mg, 77.00 μmol) and ammonia in methanol (33.0 μL, 7.0 mol / L) were dissolved in methanol (5.0 mL). Raney nickel (903.8 μg, 15.40 μmol) was added under nitrogen atmosphere, and the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. Raney nickel was removed by filtration with celite, and the filtrate was dried with anhydrous sodium sulfate and concentrated under reduced pressure to give the crude title compound (50 mg), which was used directly in the next step.

[0350] Step four: Synthesis of methyl (2S)-2,5-bis((2R)-3-amino-2-((E)-4,8-dimethyl- nona-3,7-dien-1-yl)-5-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanoate

[0351] Sodium hydroxide (22.7 mg, 567.44 μmol) was added to the crude product of Step four above dissolved in methanol (2.0 mL) and water (1.0 mL), and the reaction was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure to remove methanol, and the pH value was adjusted to pH = 6 with 2M HC1 solution. The residue was prepared by prep-HPLC [column: C-18 (spherical 40-60 μm 100A 40 g); mobile phase: [water (0.1% formic acid) - acetonitrile]; gradient: 0% - 50%] to give the target compound 009 (4.0 mg).

[0352] MS (ESI): m / z = 867.4 [M+H] + .

[0353] 1H NMR (400 MHz, DMSO-d6) δ 6.73 - 6.58 (m, 2H), 5.39 - 5.28 (m, 1H), 5.18 - 4.94 (m, 4H), 4.62 - 4.33 (m, 3H), 4.27 - 3.98 (m, 4H), 3.47 (s, 2H), 3.09 (s, 2H), 2.81 (s, 2H), 2.13 - 1.85 (m, 13H), 1.68 - 1.57 (m, 8H), 1.56 - 1.44 (m, 11H), 1.33 - 1.20 (m, 10H), 1.20 - 1.14 (m, 3H).

[0354] Example 10: Synthesis of (S)-2,5-bis((2R,3S)-3-acetoxy-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanoic acid (Compound 010)

[0355] Step one: Synthesis of (S)-2,5-bis((2R,3S)-3,5-diacetoxy-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid

[0356] Compound 1-5 (50 mg, 57.53 μmol) and triethylamine (46.57 mg, 460.25 μmol) were added into dichloromethane (2 mL). Acetyl chloride (27.10 mg, 345.19 μmol) was added under ice water bath. The reaction was stirred at room temperature for 18 hours. LCMS showed the reaction was completed. 10 mL water was added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL*3), the organic phase was combined, washed with saturated brine (5 mL*2), dried over anhydrous sodium sulfate, concentrated, and purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent] to give the title compound 10-1 (20 mg) after lyophilization.

[0357] MS (ESI): m / z = 1037.5 [M+H] + .

[0358] Step two: Synthesis of (S)-2,5-bis((2R,3S)-3-acetoxy-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid

[0359] Compound 10-1 (23 mg, 21.88 μmol) was added to methanol (1 mL), potassium phosphate (11.61 mg, 54.70 μmol) was added at 0 °C, the reaction was stirred at room temperature for 20 min. LCMS showed the reaction was completed. 3 mL pure water was added to the system, dichloromethane was used to extract (2 mL*3), the combined organic phase was washed with saturated brine (5 mL*2), dried over anhydrous sodium sulfate, concentrated, purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent], after lyophilization, the title compound 010 (5 mg) was obtained.

[0360] MS (ESI): m / z = 953.5 [M+H] + .

[0361] 1 H NMR (400 MHz, DMSO-d6) δ 9.95 (s, 2H), 6.69-6.65 (m, 2H), 5.12-4.96 (m, 6H), 4.53 (s, 1H), 4.47-4.32 (m, 1H), 4.25-4.11 (m, 3H), 3.49-3.43 (m, 2H), 2.99-2.87 (m, 2H), 2.66-2.58 (m, 2H), 2.13-1.79 (m, 20H), 1.62-1.40 (m, 24H), 1.33-1.25 (m, 6H).

[0362] Example 11: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3- (formyloxy)-5-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid (Compound 011)

[0363] In a reaction vial, N,N-dimethylformamide (1 mL) was added and put into an ice water bath, phosphorus oxychloride (105 mg, 690 μmol) was added slowly dropwise, stirred for 0.5 hour. Compound 1-5 (100 mg, 115 μmol) was dissolved in N,N-dimethylformamide (1 mL) and added slowly dropwise into the reaction system. The reaction was stirred at room temperature for 1 hour, and LCMS was used to detect the end of the reaction. The reaction was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 40%-70%) as eluent], and the title compound 011 (5 mg) was obtained after lyophilization.

[0364] MS (ESI): m / z = 925.2 [M+H] + ,923.3 [M-H] - .

[0365] 1 H NMR (400 MHz, DMSO-d6) δ 1.26-1.19 (m, 6H), 1.38-1.62 (m, 24H), 1.66-1.85 (m, 6H), 1.85-1.95 (m, 5H), 1.95-2.09 (m, 4H), 2.59-2.66 (m, 2H), 2.84-2.92 (m, 2H), 3.40 (t, J = 6.4 Hz, 2H), 4.07-4.19 (m, 3H), 4.29 (d, J = 16.4 Hz, 1H), 4.50-4.59 (m, 1H), 4.91-5.00 (m, 4H), 5.10-5.15 (m, 2H), 6.60 (s, 1H), 6.63 (s, 1H), 8.218 (s, 1H), 8.224 (s, 1H), 9.96 (br. s, 2H).

[0366] Example 12: Synthesis of (S)-2,5-bis((R)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5- hydroxy-3-(hydroxyimino)-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid (Compound 012)

[0367] Compound 9-3 (30.00 mg, 33.00 μmol) was dissolved in tetrahydrofuran (1.0 mL) and water (1.0 mL). Under nitrogen atmosphere, lithium hydroxide monohydrate (6.92 mg, 165.00 μmol) was added, and the reaction was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure to remove methanol, and the pH value was adjusted to pH = 6 with 2M HC1 solution, concentrated under reduced pressure, to give the crude product, which was purified by prep-HPLC [column: C-18 (spherical 40-60 μm 100A 40g); mobile phase: [water (0.1% formic acid) - acetonitrile]; gradient: 0% - 50%] to give the target compound 012 (2.0 mg).

[0368] MS (ESI): m / z = 895.3 [M+H] + .

[0369] 1H NMR (400 MHz, DMSO-d6) δ 11.18 (d, J = 2.3 Hz, 2H), 10.06 (s, 2H), 6.73 (d, J = 5.7 Hz, 2H), 5.32 (t, J = 4.9 Hz, 1H), 5.08 - 4.94 (m, 4H), 4.58 - 4.40 (m, 3H), 4.21 (s, 2H), 4.13 (d, J = 16.5 Hz, 1H), 3.72 - 3.67 (m, 1H), 3.66 - 3.61 (m, 1H), 3.51 - 3.44 (m, 2H), 2.04 - 1.90 (m, 10H), 1.90 - 1.81 (m, 4H), 1.76 - 1.65 (m, 3H), 1.62 - 1.53 (m, 8H), 1.52 - 1.37 (m, 18H).

[0370] Example 13: Synthesis of (2S)-2,5-bis((2R,3S)-2-(4,8-dimethylnonyl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid (Compound 013)

[0371] Compound 1-5 (50 mg, 57.53 μmol), palladium on carbon (5 mg, 4.12 μmol, 10% purity), palladium hydroxide on carbon (5 mg, 3.56 μmol, 10% purity) were added into methanol (1 mL), the reaction solution was stirred at 20-30 °C for 36 hours under hydrogen atmosphere. LCMS showed the reaction was completed, the reaction solution was filtered, purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent], after lyophilization, the title compound 013 (15 mg) was obtained.

[0372] MS (ESI): m / z = 877.5 [M+H] + .

[0373] 1 H NMR (400 MHz, DMSO-d6) δ 9.80 (s, 2H), 6.70 - 6.58 (m, 2H), 5.14 (s, 2H), 4.61 - 4.50 (m, 1H), 4.37 - 4.25 (m, 1H), 4.14 (s, 2H), 4.11 - 4.00 (m, 1H), 3.79 - 3.67 (m, 2H), 3.49 - 3.42 (m, 2H), 2.87 - 2.75 (m, 2H), 2.47 - 2.37 (m, 2H), 1.99 - 1.86 (m, 1H), 1.75 (s, 1H), 1.67 - 1.32 (m, 14H), 1.32 - 0.95 (m, 23H), 0.87 - 0.74 (m, 18H).

[0374] Example 14: Synthesis of (S)-2,5-bis((2R,3S)-6-bromo-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid (Compound 014)

[0375] Compound 1-5 (100 mg, 0.12 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and stirred at -20 °C for 10-20 min, then a solution of N-bromosuccinimide (51 mg, 0.29 mmol) in tetrahydrofuran (5 mL) was added dropwise, and the dropwise addition was completed within 1 h. The reaction was monitored by LC-MS. A 10% sodium sulfite aqueous solution (4 mL) was added to the reaction mixture at -20 °C, and stirred for 10 min, then moved to room temperature, and dichloromethane (20 mL) and water (10 mL) were added, and the organic phase was concentrated to dryness. The residue was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 55%-75%) as eluent], and the title compound 014 (17 mg) was obtained after lyophilization.

[0376] MS (ESI): m / z = 1025.0, 1027.0 [M+H] + .

[0377] 1 H NMR (400 MHz, DMSO-d6) δ 1.21 - 1.13 (m, 6H), 1.50 - 1.62 (m, 23H), 1.72 - 1.84 (m, 1H), 1.86 - 1.94 (m, 5H), 1.95 - 2.02 (m, 4H), 2.04 - 2.14 (m, 4H), 2.54 - 2.62 (m, 2H), 2.89 (dt, J = 18.1, 5.4 Hz, 2H), 3.41 - 3.51 (m, 2H), 3.75 (dd, J = 7.1, 13.1 Hz, 2H), 4.08 (d, J = 17.2 Hz, 1H), 4.14 (s, 2H), 4.31 (d, J = 16.7 Hz, 1H), 4.50 - 4.62 (m, 1H), 5.00 - 5.06 (m, 2H), 5.07 - 5.14 (m, 2H), 5.21 (d, J = 3.9 Hz, 2H).

[0378] Example 15: Synthesis of (S)-2,5-bis((2R,3S)-6-((dimethylamino)methyl)-2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanoic acid (Compound 015)

[0379] Compound 1-5 (100 mg, 0.12 mmol), 40% aqueous dimethylamine (45.5 mg, 0.40 mmol), aqueous formaldehyde (22.5 mg, 0.29 mmol, 40% formaldehyde content) were added to a mixture of tetrahydrofuran (2.5 ml) and water (2.5 ml) at room temperature and stirred at room temperature for 12 hours. LC-MS showed the reaction was completed. The reaction solution was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 55%-85%) as eluent] and the title compound 015 (25 mg) was obtained after lyophilization.

[0380] MS (ESI): m / z = 983.4 [M+H] + .

[0381] 1 H NMR (400 MHz, DMSO-d6) δ 1.20 - 1.12 (m, 6H), 1.40 - 1.65 (m, 24H), 1.70 - 1.80 (m, 1H), 1.86 - 1.95 (m, 5H), 1.95 - 2.05 (m, 4H), 2.05 - 2.14 (m, 4H), 2.27 (d, J = 3.3 Hz, 12H), 2.42 - 2.48 (m, 2H), 2.82 (dt, J = 17.6, 5.2 Hz, 2H), 3.45 (t, J = 7.4 Hz, 2H), 3.73 (dd, J = 7.1, 13.1 Hz, 2H), 4.07 - 4.27 (m, 8H), 4.65 (dd, J = 4.6, 10.4 Hz, 1H), 5.00 - 5.07 (m, 2H), 5.07 - 5.16 (m, 2H).

[0382] Example 16: Synthesis of (S)-2,5-bis((2R,3S)-5-((diethylglycyl)oxy)-2-(((E)-4,8- dimethylnona-3,7-dien-1-yl)-3-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanoic acid (Compound 016)

[0383] Step one: Synthesis of diethylglycyl chloride

[0384] Compound 16-1 (200 mg, 1.52 mmol) was added into dichloromethane (3 mL), oxalyl chloride (290.29 mg, 2.29 mmol) and 1 drop of N,N-dimethylformamide were added under ice water bath. The reaction was stirred at room temperature for 3 hours. The reaction was concentrated to get the crude title compound 16-2 (300 mg), which was used directly for the next step.

[0385] Step two: synthesis of (S)-2,5-bis((2R,3S)-5-((diethylglycyl)oxy)-2-(((E)-4,8- dimethylnona-3,7-dien-1-yl)-3-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E] isoindol-8(2H)-yl)pentanoic acid

[0386] Compound 16-2 (51.64 mg, 345.19 μmol) and triethylamine (145.54 mg, 1.44 mmol) were added into dichloromethane (4 mL), compound 1-5 (50.00 mg, 57.53 μmol) was added under ice water bath. The reaction was stirred at room temperature for 18 hours. LCMS showed the reaction was completed. The reaction was quenched by adding water (20 mL) under ice water bath, extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent], the title compound 016 (10 mg) was obtained after lyophilization.

[0387] MS (ESI): m / z = 1095.8 [M+H] + .

[0388] 1H NMR (400 MHz, DMSO-d6) δ 6.95 (d, J = 16.1 Hz, 2H), 5.35 - 5.28 (m, 2H), 5.13 - 5.08 (m, 2H), 5.08 - 4.99 (m, 2H), 4.72 (s, 1H), 4.44 - 4.25 (m, 4H), 3.83 - 3.72 (m, 2H), 3.67 - 3.62 (m, 4H), 3.55 - 3.48 (m, 2H), 2.82 - 2.70 (m, 2H), 2.72 - 2.61 (m, 8H), 2.14 - 2.07 (m, 4H), 2.03 - 1.86 (m, 10H), 1.63 - 1.50 (m, 24H), 1.20 (d, J = 14.1 Hz, 6H), 1.08 - 0.99 (m, 12H).

[0389] Example 17: Synthesis of (S)-2,5-bis((2R,3S)-5-amino-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanoic acid (Compound 017)

[0390] Step one: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5-(((trifluoromethyl)sulfonyl)oxy)-3-hydroxy-2-methyl-7-oxo-3,4,7,9- tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanoic acid

[0391] Compound 17-1 (249.61 mg, 920.50 umol) was dissolved in N,N- dimethylformamide, potassium carbonate (190.54 mg, 1.38 mmol) and compound 1-5 (200 mg, 230.13 umol) were added successively. The reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was complete. The reaction was subjected to prep-HPLC [column: C-18 (spherical 40-60 pm 100 A 40 g); mobile phase: [water (0.1% formic acid) - acetonitrile]; gradient: 0%-50%] to give the target compound 17-2 (90 mg).

[0392] MS (ESI): m / z = 1133.1 [M+H] + .

[0393] Step 2: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-5- ((diphenylmethylene)amino)-3-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E] isoindol-8(2H)-yl)pentanoic acid

[0394] Compound 17-2 (50 mg, 44.12 μmol), benzophenone imine (79.96 mg, 441.22 μmol, 74.04 μL) and potassium carbonate (30.49 mg, 220.61 μmol) were dissolved in dioxane (0.50 mL), and 9,9-dimethyl-4,5-bisdiphenylphosphine xanthene (2'-amino-1,1'- biphenyl-2-yl)palladium(II) (4.19 mg, 4.41 μmol) was added under N2protection. The reaction was stirred at 100 °C for 16 h under nitrogen atmosphere. LCMS showed the reaction was complete. The reaction was filtered and concentrated under reduced pressure, and the crude product was purified by flash silica gel column chromatography [column: Sepax FCI 4.6 x 50 mm, 5 μm; mobile phase: 0-15% methanol in dichloromethane at a flow rate of 18 mL / min] to give the title compound 17-3 (52 mg). 4g flash silica gel column, gradient 0-15% methanol in dichloromethane at a flow rate of 18 mL / min] to give the title compound 17-3 (52 mg).

[0395] MS (ESI): m / z = 598.8 [M / 2+H] + .

[0396] Step 3: Synthesis of (S)-2,5-bis((2R,3S)-5-amino-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E] isoindol-8(2H)-yl)pentanoic acid

[0397] Compound 17-3 (40 mg, 33.46 μmol), hydroxylamine hydrochloride (9.30 mg, 133.83 μmol) and sodium acetate (27.45 mg, 334.58 μmol) were dissolved in methanol (1.00 mL), and the reaction was stirred at room temperature for 16 h. LCMS showed the reaction was complete. The reaction was filtered and concentrated under reduced pressure, and the crude product was purified by prep-HPLC [column: Phenomenex Gemini NX 150 x 30 mm, 5 μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 57%-97%] to give the title compound 017 (6.4 mg).

[0398] MS (ESI): m / z = 867.3 [M+H] + .

[0399] 1H NMR (400 MHz, DMSO-d6) δ 6.54 (s, 1H), 6.50 (s, 1H), 5.21 - 4.89 (m, 10H), 4.71 (br dd, J = 5.0, 10.1 Hz, 1H), 4.24 - 4.08 (m, 4H), 3.76 (quin, J = 6.1 Hz, 2H), 3.47 (br t, J = 6.7 Hz, 2H), 2.67 (td, J = 5.9, 17.1 Hz, 2H), 2.38 - 2.29 (m, 2H), 2.15 - 2.05 (m, 4H), 2.04 - 1.97 (m, 4H), 1.95 - 1.88 (m, 5H), 1.65 - 1.57 (m, 10H), 1.54 (s, 14H), 1.20 - 1.10 (m, 6H).

[0400] Example 18: Synthesis of (S)-2,5-bis((2R,3S)-5-(difluoromethoxy)-2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3-hydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanoic acid (Compound 018)

[0401] Compound 1-5 (60 mg, 69.04 μmol) and potassium hydroxide (77.47 mg, 1.38 mmol) were added into dichloromethane (1 mL) and deionized water (0.6 mL). The reaction solution was added into difluorobromomethyl trimethylsilane (84.13 mg, 414.23 μmol) at -78 °C. The reaction solution was stirred at room temperature for 8 hours. LCMS showed the reaction was completed. The reaction solution was cooled and poured into water (20 mL), extracted with ethyl acetate (30 mL*3), the combined organic layers were washed with water (40 mL*2) and saturated brine (40 mL) successively, dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, the residue was purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60% - 90%) as eluent] and the title compound 018 (10 mg) was obtained after lyophilization.

[0402] MS (ESI): m / z = 969.5 [M+H] + .

[0403] 1H NMR (400 MHz, DMSO-d6) δ 7.56 - 7.09 (m, 2H), 6.99 - 6.93 (m, 2H), 5.34 - 5.29 (m, 2H), 5.13 - 5.07 (m, 2H), 5.07 - 4.99 (m, 2H), 4.59 - 4.42 (m, 2H), 4.29 (s, 2H), 4.18 (d, J = 17.6 Hz, 1H), 3.82 - 3.77 (m, 2H), 3.54 - 3.45 (m, 2H), 2.97 - 2.85 (m, 2H), 2.67 - 2.54 (m, 2H), 2.18 - 1.83 (m, 14H), 1.79 - 1.39 (m, 24H), 1.21 (d, J = 8.8 Hz, 6H).

[0404] Example 19: Synthesis of 2,2'-(((2R,2'R,3S,3'S)-((S)-1-carboxybutane-1,4-diyl)bis(2- ((E)-4,8-dimethylnona-3,7-dien-1-yl)-3-hydroxy-2-methyl-7-oxo-2,3,4,7,8,9- hexahydropyrano[2,3-E]isoindol-8,5-diyl))bis(oxy))diacetic acid (Compound 019)

[0405] Step one: Synthesis of diethyl 2,2'-(((2R,2'R,3S,3'S)-((S)-5-(2-ethoxy-2-oxoethoxy)-5- oxopentane-1,4-diyl)bis(2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3-hydroxy-2- methyl-7-oxo-2,3,4,7,8,9-hexahydropyrano[2,3-E]isoindol-8,5-diyl))bis(oxy))diacetate

[0406] Compound 1-5 (150 mg, 0.17 mmol) was dissolved in N,N-dimethylformamide (4.5 mL), then sodium iodide (103 mg, 0.68 mmol), potassium carbonate (119 mg, 0.86 mmol) and ethyl bromoacetate (115 mg, 0.69 mmol) were added. The reaction was stirred at room temperature for 12 hours, then LCMS showed the reaction was completed. Ethyl acetate (10 mL) and water (10 mL) were added to the reaction, then the organic phase was concentrated to get the crude product of title compound 19-1, which was used directly for the next step.

[0407] MS (ESI): m / z = 1127.5 [M+H] + .

[0408] Step two: synthesis of 2,2'-(((2R,2'R,3S,3'S)-((S)-1-carboxybutane-1,4-diyl)bis(2-((E)-4,8- dimethylnona-3),7-dien-1-yl)-3-hydroxy-2-methyl-7-oxo-2,3,4,7,8,9-hexahydropyrano[2,3- E]isoindol-8,5-diyl))bis(oxy))diacetic acid

[0409] The concentrate prepared in Step one (compound 19-1 crude) was dissolved in tetrahydrofuran (4 mL), and a solution of lithium hydroxide monohydrate (90.63 mg, 2.16 mmol) in water (1 mL) was added. The reaction was stirred at room temperature for 1.5 hours. LCMS showed the reaction was completed. The reaction was adjusted to pH ≈ 7 with acetic acid, and purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 55%-85%) as eluent]. Compound 019 (9 mg) was obtained after lyophilization.

[0410] MS (ESI): m / z = 985.7 [M+H] + .

[0411] 1 H NMR (400 MHz, DMSO-d6) δ 1.21 - 1.13 (m, 6H), 1.50 - 1.62 (m, 24H), 1.70 - 1.80 (m, 1H), 1.86 - 1.94 (m, 5H), 1.95 - 2.04 (m, 4H), 2.06 - 2.15 (m, 4H), 2.51 - 2.57 (m, 2H), 2.89 (dt, J = 17.2, 4.8 Hz, 2H), 3.47 (t, J = 6.1 Hz, 2H), 3.75 (dd, J = 5.9, 11.9 Hz, 2H), 4.09 - 4.24 (m, 3H), 4.32 - 4.43 (m, 5H), 4.57 (dd, J = 10.7, 4.4 Hz, 1H), 5.00 - 5.16 (m, 6H), 6.53 (s, 1H), 6.55 (s, 1H).

[0412] Example 20: Synthesis of (2S)-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8-(2H)-yl)pentanamido)-3,3,3-trifluoropropionic acid & (2R)-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9- tetrahydropyrano[2,3-e]isoindol-8-(2H)-yl)pentanamido)-3,3,3-trifluoropropionic acid (Compound 048 & 049)

[0413] Compound 3-1 (110 mg, 114 pmol), trifluoropropanoic acid hydrochloride (62 mg, 345 pmol), 4-dimethylaminopyridine (14 mg, 115 pmol) were dissolved in dry N,N-dimethylformamide (2 mL), N,N-diisopropylethylamine (120 pL, 690 pmol) was added slowly to the reaction, the reaction was stirred at room temperature for 2 hours. LC-MS showed the reaction was completed. Saturated brine (10 mL) was added to the reaction, extracted with ethyl acetate (10 mL) and dried over anhydrous sodium sulfate to give the crude product. The crude product was purified by prep-HPLC [YMC-TAR column 5 pm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 7 mmol / L NH4HCO3) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 30%-60%] to give title compounds 048 (10 mg) and 049 (9 mg).

[0414] Compound 048:

[0415] HPLC [column: Agilent Poroshell 120 EC-C18 (4,6*100, 2.7 pm); mobile phase: 0.1% formic acid-acetonitrile / 0.1% formic acid-water, gradient (acetonitrile gradient ratio 10%-95%) elution for 18 minutes, isocratic (acetonitrile gradient ratio 10%) elution for 5 minutes; flow rate 1.0 mL / min] retention time: 13.329 min

[0416] MS (ESI): m / z = 994.3 [M+H] + .

[0417] 1H NMR (400 MHz, DMSO-d6) δ 9.88 - 9.68 (m, 2H), 8.14 (s, 1H), 6.68 - 6.60 (m, 2H), 5.20 - 4.99 (m, 6H), 4.92 (d, J = 10.2, 5.2 Hz, 1H), 4.50 - 4.65 (m, 1H), 4.32 (d, J = 16.8 Hz, 1H), 4.25 - 4.08 (m, 3H), 3.80 - 3.68 (m, 2H), 3.52 - 3.44 (m, 2H), 2.89 - 2.76 (m, 2H), 2.48 - 2.39 (m, 2H), 2.16 - 2.05 (m, 4H), 2.05 - 1.95 (m, 4H), 1.95 - 1.88 (m, 4H), 1.88 - 1.70 (m, 2H), 1.67 - 1.42 (m, 24H), 1.20 - 1.12 (m, 6H).

[0418] Compound 049:

[0419] HPLC [Column: Agilent Poroshell 120 EC-C18 (4,6*100, 2.7 pm); Mobile phase: 0.1% formic acid-acetonitrile / 0.1% formic acid-water, gradient (acetonitrile gradient ratio 10% - 95%) elution for 18 minutes, isocratic (acetonitrile gradient ratio 10%) elution for 5 minutes; Flow rate 1.0 mL / min] Retention time: 14.550 min

[0420] MS (ESI): m / z = 994.3 [M+H] + .

[0421] 1 H NMR (400 MHz, DMSO-d6) δ 9.88 - 9.68 (m, 2H), 8.36 (s, 1H), 6.68 - 6.60 (m, 2H), 5.20 - 5.02 (m, 6H), 4.98 (d, J = 9.6, 5.8 Hz, 1H), 4.68 - 4.53 (m, 1H), 4.46 (d, J = 17.0 Hz, 1H), 4.25 - 4.10 (m, 3H), 3.80 - 3.68 (m, 2H), 3.52 - 3.44 (m, 2H), 2.88 - 2.77 (m, 2H), 2.48 - 2.39 (m, 2H), 2.16 - 2.05 (m, 4H), 2.05 - 1.96 (m, 4H), 1.96 - 1.88 (m, 4H), 1.81 - 1.70 (m, 2H), 1.65 - 1.48 (m, 24H), 1.19 - 1.11 (m, 6H).

[0422] Example 21: Synthesis of (R)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido)-3-fluoropropionic acid (Compound 050)

[0423] Step one: Synthesis of (R)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido)-3-fluoropropionic acid methyl ester

[0424] Compound 1-5 (50.00 mg, 57.53 μmol), (R)-2-amino-3-fluoropropionic acid methyl ester hydrochloride (10.88 mg, 69.04 μmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (43.41 mg, 114.16 μmol) and N,N-diisopropylethylamine (29.74 mg, 230.13 μmol) were dissolved in N,N-dimethylformamide (1.0 mL), and the reaction solution was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was quenched by adding water at 0 °C, and then extracted with ethyl acetate (10.0 mL*3). The organic phase was washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the title compound 21-1 (30.00 mg).

[0425] MS (ESI): m / z = 972.50 [M+H] + .

[0426] Step two: Synthesis of (R)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido)-3-fluoropropionic acid

[0427] Lithium hydroxide (7.39 mg, 308.58 μmol) and compound 21-1 (30.00 mg, 30.86 μmol) were dissolved in a tetrahydrofuran (1.0 mL) and water (0.5 mL) solution, the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure, the pH value was adjusted to pH = 6 with 2M HCl solution, concentrated under reduced pressure, the residue was purified by Prep-HPLC [YMC-TAR column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.05% formic acid) and acetonitrile with decreasing polarity as eluents; acetonitrile gradient ratio 80%-100%] to give the target compound 050 (9.31 mg).

[0428] MS (ESI): m / z = 958.40 [M+H] + .

[0429] 1 H NMR (400 MHz, DMSO-d6) δ 9.86 - 9.66 (m, 2H), 8.35 (s, 1H), 6.65 (s, 1H), 6.62 (s, 1H), 5.19 - 5.08 (m, 4H), 5.08 - 5.00 (m, 2H), 4.88 - 4.71 (m, 2H), 4.41 (d, J = 17.0 Hz, 1H), 4.21 - 4.12 (m, 3H), 3.79 - 3.69 (m, 2H), 3.63 - 3.43 (m, 4H), 2.82 (dt, J = 17.4, 5.4 Hz, 2H), 2.47 - 2.42 (m, 2H), 2.15 - 2.06 (m, 4H), 2.04 - 1.96 (m, 4H), 1.94 - 1.88 (m, 4H), 1.85 - 1.74 (m, 2H), 1.65 - 1.56 (m, 10H), 1.55 - 1.50 (m, 11H), 1.19 - 1.12 (m, 6H).

[0430] Example 22: Synthesis of (S)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentanamido)-3-fluoropropionic acid (Compound 051)

[0431] Step one: synthesis of (S)-methyl 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanamido)-3-fluoropropionate

[0432] Compound 1-5 (50.00 mg, 57.53 μmol), (S)-methyl 2-amino-3-fluoropropionate hydrochloride (10.88 mg, 69.04 μmol), 2-(7-azobenzotriazole)-N,N,N',N'- tetramethyluronium hexafluorophosphate (43.41 mg, 114.16 μmol) and N,N- diisopropylethylamine (29.74 mg, 230.13 μmol) were dissolved in N,N- dimethylformamide (1.0 mL), the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was quenched by adding water at 0 °C, then extracted with ethyl acetate (10.0 mL*3), the organic phase was washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound 22-1 (40.00 mg).

[0433] MS (ESI): m / z = 972.50 [M+H] + .

[0434] Step two: synthesis of (S)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- E]isoindol-8(2H)-yl)pentanamido)-3-fluoropropionic acid

[0435] Lithium hydroxide (9.85 mg, 411.44 μmol) and compound 22-1 (40.00 mg, 41.14 μmol) were dissolved in a tetrahydrofuran (1.0 mL) and water (0.5 mL) solution, the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure, the pH value was adjusted to 6 with 2M HCl solution, concentrated under reduced pressure, the residue was purified by Prep-HPLC [YMC-TAR column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.05% formic acid) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 80%-100%] to give the target compound 051 (9.31 mg).

[0436] MS (ESI): m / z = 958.40 [M+H] + .

[0437] 1 H NMR (400 MHz, DMSO-d6) δ 9.80 - 9.70 (m, 2H), 8.42 (t, J = 5.8 Hz, 1H), 6.65 (s, 1H), 6.62 (s, 1H), 5.19 - 5.08 (m, 4H), 5.08 - 5.00 (m, 2H), 4.96 - 4.75 (m, 2H), 4.41 (d, J = 17.0 Hz, 1H), 4.23 - 4.12 (m, 3H), 3.74 (q, J = 6.0 Hz, 2H), 3.65 - 3.39 (m, 5H), 2.88 - 2.75 (m, 2H), 2.47 - 2.37 (m, 2H), 2.17 - 2.05 (m, 4H), 2.03 - 1.96 (m, 4H), 1.94 - 1.88 (m, 4H), 1.84 - 1.74 (m, 2H), 1.64 - 1.57 (m, 9H), 1.55 - 1.44 (m, 14H), 1.19 - 1.12 (m, 6H).

[0438] Example 23: Synthesis of (2S)-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8-(2H)-yl)pentanamido)- 3,3-difluoropropionic acid & (2R)-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8-(2H)-yl)pentanamido)-3,3- difluoropropionic acid (Compound 052 & 053)

[0439] Step one: Synthesis of tert-butyl 2-((diphenylmethylene)amino)-3,3-difluoropropanoate

[0440] Compound 23-1 (4.5 g, 15.23 mmol) was dissolved in tetrahydrofuran (100 mL) under nitrogen protection, and potassium tert-butoxide (1 M tetrahydrofuran solution, 18.28 mL) was added dropwise at 0 °C. After stirring at 0 °C for 30 min, compound 23-2 (3.66 g, 18.28 mmol, 2.31 mL) was added. The reaction solution was stirred at 0 °C for 2 h. LCMS showed that the product was generated. The reaction solution was added with ethyl acetate (200 mL), and then washed with saturated ammonium chloride solution (50 mL). The organic phase was dried with anhydrous sodium sulfate and concentrated. The crude product was purified by flash silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give compound 23-3 (4.5 g, 12.23 mmol, 80%). 80g Fast silica gel column, gradient 0-5% petroleum ether / ethyl acetate, flow rate 60 mL / min) to give the crude product. Further purification by preparative high performance liquid chromatography (column: Boston Prime C18 150*30 mm*5 μm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 55%-95%) to give the title compound 23-3 (0.43 g).

[0441] 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.79-7.65 (m, 2H), 7.57-7.34 (m, 6H), 7.27-7.19 (m, 2H), 6.48-6.02 (m, 1H), 4.31 (s, 1H), 1.48 (s, 9H).

[0442] Step two: synthesis of tert-butyl 2-((tert-butoxycarbonyl)amino)-3,3- difluoropropanoate

[0443] Compound 23-3 (260 mg, 752.79 μmol) was dissolved in tetrahydrofuran (3 mL), (Boc)20 (246.44 mg, 1.13 mmol) and palladium on carbon (52.00 mg, 10% content) were added. The reaction was stirred at 25 °C for 16 hours under hydrogen balloon atmosphere (15 psi). The reaction was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography (gradient 0-5% petroleum ether / ethyl acetate, flow rate 35 mL / min) to give the title compound 23-4 (170 mg). 12g Fast silica gel column, gradient 0-5% petroleum ether / ethyl acetate, flow rate 60 mL / min) to give the crude product. Further purification by preparative high performance liquid chromatography (column: Boston Prime C18 150*30 mm*5 μm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 55%-95%) to give the title compound 23-3 (0.43 g).

[0444] 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.79-7.65 (m, 2H), 7.57-7.34 (m, 6H), 7.27-7.19 (m, 2H), 6.48-6.02 (m, 1H), 4.31 (s, 1H), 1.48 (s, 9H).

[0445] Step three: synthesis of 2-amino-3,3-difluoropropanoic acid hydrochloride

[0446] Compound 23-4 (170 mg, 604.35 μmol) was dissolved in 2M hydrochloric acid in dioxane (2 mL), and the reaction was stirred at 25 °C for 32 hours. The reaction was concentrated under reduced pressure to give the title compound 23-5 (97.62 mg).

[0447] 1H NMR (400 MHz, DMSO-d6) δ 9.95 - 8.14 (m, 2H), 6.86 - 6.29 (m, 1H), 4.84 - 4.61 (m, 1H)

[0448] Step four: synthesis of (2S)-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8-(2H)-yl)pentanamido)- 3,3-difluoropropionic acid & (2R)-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8-(2H)-yl)pentanamido)-3,3- difluoropropionic acid

[0449] Compound 23-5 (83.26 mg, 515.44 μmol) and compound 3-1 (166 mg, 171.81 μmol) were dissolved in N,N-dimethylformamide, N,N-diisopropylethylamine (111.03 mg, 859.07 μmol) and N,N-dimethylaminopyridine (20.99 mg, 171.81 μmol) were added. The reaction was stirred at 25 °C for 16 hours. LCMS showed the reaction was completed. The reaction was purified by preparative high performance liquid chromatography (column: Boston Prime C18 150*30mm*5μm; mobile phase: [water(0.05% ammonia water + 0.001% ammonium bicarbonate)-acetonitrile]; gradient: 37%-77%) to give two products. The two products were further purified by preparative high performance liquid chromatography (column: Boston Prime C18 150*30mm*5μm; mobile phase: [water(0.225% formic acid)-acetonitrile]; gradient: 53%-93%) to give compound 052 (2.2 mg) and compound 053 (4.8 mg).

[0450] Compound 052:

[0451] LC-MS [column: NanoChrom ChromCore 120 C18 3μm, 3.0*30mm; mobile phase: Phase A: 0.0375% trifluoroacetic acid-water, Phase B: 0.0187% trifluoroacetic acid-acetonitrile (B phase gradient ratio 30%-90%), elution for 6 minutes; flow rate 0.8 mL / min] retention time: 4.479 min.

[0452] MS (ESI): m / z = 976.6 [M+H] + .

[0453] 1 H NMR (400 MHz, DMSO-d6) δ 9.82 - 9.74 (m, 2H), 8.62 - 8.42 (m, 1H), 8.70 - 8.32 (m, 1H), 6.68 - 6.60 (m, 2H), 6.47 - 6.07 (m, 1H), 5.24 - 4.93 (m, 7H), 4.75 - 4.59 (m, 1H), 4.77 - 4.56 (m, 1H), 4.48 - 4.38 (m, 1H), 4.29 - 4.09 (m, 3H), 3.82 - 3.72 (m, 2H), 2.88 - 2.76 (m, 2H), 2.18 - 1.79 (m, 15H), 1.66 - 1.44 (m, 26H), 1.20 - 1.12 (m, 6H).

[0454] Compound 053:

[0455] LC-MS [Column: NanoChrom ChromCore 120 C18 3 pm, 3.0*30 mm; mobile phase: Phase A: 0.0375% trifluoroacetic acid - water, Phase B: 0.0187% trifluoroacetic acid - acetonitrile (gradient ratio of Phase B 30% - 90%), elution for 6 min; flow rate 0.8 mL / min] Retention time: 4.628 min.

[0456] MS (ESI): m / z = 976.6 [M+H] + .

[0457] 1 H NMR (400 MHz, DMSO-d6) δ 9.82 - 9.74 (m, 2H), 8.62 - 8.42 (m, 1H), 8.70 - 8.32 (m, 1H), 6.68 - 6.60 (m, 2H), 6.47 - 6.07 (m, 1H), 5.24 - 4.93 (m, 7H), 4.75 - 4.59 (m, 1H), 4.77 - 4.56 (m, 1H), 4.48 - 4.38 (m, 1H), 4.29 - 4.09 (m, 3H), 3.82 - 3.72 (m, 2H), 2.88 - 2.76 (m, 2H), 2.18 - 1.79 (m, 15H), 1.66 - 1.44 (m, 26H), 1.20 - 1.12 (m, 6H).

[0458] Example 24: Synthesis of 2,2'-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnon-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoyl)azanediyl)diacetic acid (Compound 054)

[0459] Step one: synthesis of diethyl 2,2'-(((S)-2,5-bis((2R,3S)-2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9- tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)azanediyl)diacetate

[0460] Compound 1-3 (100 mg, 115 pmol), diethyl iminodiacetate (65.20 mg, 345 pmol), 2-(7-azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65 mg, 171 pmol) and N,N-diisopropylethylamine (44.61 mg, 345 pmol) were dissolved in N,N-dimethylformamide (2 mL), the reaction was stirred at room temperature for 2 hours. 10 mL water was added at 0 °C to quench, extracted with ethyl acetate (10.00 mL*3), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound 24-1 (100 mg).

[0461] Step two: synthesis of 2,2'-(((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7- dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoyl)azanediyl)diacetic acid

[0462] Compound 24-1 (100 mg, 96.12 pmol) and lithium hydroxide (25.32 mg, 1.06 mmol) were dissolved in a mixed solvent of tetrahydrofuran (1 mL) and water (1 mL), the reaction was stirred at room temperature for 2 hours. LCMS showed that the reaction was complete. The reaction was concentrated under reduced pressure, the pH value was adjusted to pH = 6 using 2M aqueous HC1 solution, and concentrated under reduced pressure. The residue was purified by pre-HPLC [YMC-TAR column 5 pm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.1% formic acid) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 75%-90%] to give the target compound 054 (10 mg).

[0463] MS (ESI): m / z = 984.40 [M+H] + .

[0464] 1H NMR (400 MHz, DMSO-d6) δ 9.98 - 9.56 (m, 2H), 6.66 - 6.58 (m, 2H), 5.22 - 4.97 (m, 7H), 4.28 - 4.01 (m, 5H), 3.96 - 3.82 (m, 2H), 3.77 - 3.68 (m, 2H), 3.47 - 3.40 (m, 2H), 2.86 - 2.76 (m, 2H), 2.47 - 2.41 (m, 2H), 2.17 - 2.05 (m, 4H), 2.05 - 1.96 (m, 4H), 1.96 - 1.85 (m, 4H), 1.82 - 1.33 (m, 27H), 1.20 - 1.10 (m, 6H).

[0465] Example 25: Synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)-N- methylpentanamido)propanoic acid (Compound 055)

[0466] Step one: Synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)-N- methylpentanamido)propanoic acid ethyl ester

[0467] Compound 1-5 (1.00 g, 1.15 mmol), 3-(methylamino)propanoic acid ethyl ester (452.79 mg, 3.45 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (651.15 mg, 1.71 mmol) and N,N-diisopropylethylamine (594.83 mg, 4.63 mmol) were added into N,N-dimethylformamide (10 mL). The reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The reaction was concentrated and purified by RP-flash (column: Welch XB-C18, 21.2*250mm, 5um; using mixture of water (containing 7mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 40%-63%) as eluent; flow rate: 25 mL / min; ) to give the title compound 25-1 (600.00 mg).

[0468] MS (ESI): m / z = 982.5 [M+H] +

[0469] Step two: synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)-N- methylpentanamido)propanoic acid

[0470] Step two: synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)-N- methylpentanamido)propanoic acid

[0471] MS (ESI): m / z = 954.5 [M+H] + .

[0472] 1 H NMR (400 MHz, DMSO-d6) δ 9.82 - 9.72 (m, 2H), 6.65 (s, 1H), 6.62 (s, 1H), 5.16 - 5.08 (m, 5H), 5.06 - 5.01 (m, 2H), 4.19 - 4.04 (m, 4H), 3.75 - 3.70 (m, 3H), 3.48 - 3.39 (m, 3H), 2.98 (s, 1H), 2.86 - 2.74 (m, 3H), 2.47 - 2.28 (m, 4H), 2.14 - 1.87 (m, 13H), 1.75 - 1.68 (m, 3H), 1.65 - 1.40 (m, 22H), 1.20 - 1.12 (m, 6H).

[0473] Example 26: synthesis of 3-amino-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7- dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)- yl)pentanamido)propanoic acid (Compound 056)

[0474] Compound 1-5 (100 mg, 115 pmol), methyl 2-amino-3-((fluorenylmethoxycarbonyl)amino)propanoate (hydrochloride, 130 mg, 345 pmol), 2-(7-azobenzotriazol-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65.11 mg, 171.23 pmol) and N,N-diisopropylethylamine (45 mg, 345 pmol) were dissolved in N,N-dimethylformamide (2 mL), the reaction was stirred at room temperature for 2 hours. 10 mL water was added at 0 °C to quench, extracted with ethyl acetate (10.00 mL*3), the organic phase was washed with saturated brine (10 mL), concentrated under reduced pressure. The concentrate and lithium hydroxide (25.32 mg, 1.06 mmol) were dissolved in a mixture solvent of tetrahydrofuran (1 mL) and water (1 mL), the reaction was stirred at room temperature for 2 hours. LCMS showed that the reaction was complete. The reaction was concentrated under reduced pressure, the pH value was adjusted to pH = 7 using 2M aqueous HC1 solution, concentrated under reduced pressure, the residue was purified by pre-HPLC [YMC-TAR column 5 pm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 7 mmol / L NH4HCO3) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 50%-70%] to give the target compound 056 (23 mg).

[0475] MS (ESI): m / z = 955.3 [M+H] + .

[0476] 1 H NMR (400 MHz, DMSO-d6) d 10.49 - 9.00 (m, 2H), 7.67 (d, J = 6.1 Hz, 1H), 6.63 - 6.54 (m, 2H), 5.21 - 4.90 (m, 6H), 4.75 - 4.70 (m, 1H), 4.32 - 4.28 (m, 1H), 4.16 - 4.05 (m, 3H), 3.85 - 3.77 (m, 1H), 3.72 - 3.63 (m, 2H), 2.92 - 2.85 (m, 1H), 2.82 - 2.69 (m, 4H), 2.69 - 2.59 (m, 1H), 2.42 - 2.34 (m, 2H), 2.16 - 2.00 (m, 4H), 2.00 - 1.89 (m, 4H), 1.89 - 1.78 (m, 4H), 1.78 - 1.65 (m, 2H), 1.59 - 1.41 (m, 24H), 1.12 - 1.06 (m, 6H).

[0477] Example 27: Synthesis of 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)ethane-1- sulfonic acid (Compound 057)

[0478] Compound 3-1 (222 mg, 0.23 mmol) was dissolved in N,N-dimethylformamide (4 mL), 4-dimethylaminopyridine (25.2 mg, 0.21 mmol), N,N-diisopropylethylamine (72 mg, 0.56 mmol), taurine (86 mg, 0.69 mmol) were added successively and stirred uniformly, and temperature was controlled to 0-10 °C and stirred for 3 hours. LCMS showed that the reaction was completed. The reaction solution was filtered and purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 40%-70%) as eluent], and the title compound 057 (10 mg) was obtained after freeze-drying.

[0479] MS (ESI): m / z = 976.3 [M+H] + .

[0480] 1 H NMR (400 MHz, DMSO-d6) δ 9.82-9.63 (m, 2H), 8.26-8.18 (m, 1H), 6.67-6.59 (m, 2H), 5.19-5.05 (m, 6H), 4.78-4.68 (m, 1H), 4.45-4.36 (m, 1H), 4.20-4.12 (m, 3H), 3.75-3.64 (m, 4H), 3.50-3.43 (m, 2H), 3.20 (d, J = 6.3 Hz, 2H), 2.88-2.76 (m, 2H), 2.47-2.43 (m, 2H), 2.12-1.91 (m, 13H), 1.61-1.45 (m, 24H), 1.20-1.11 (m, 6H).

[0481] Example 28: Synthesis of 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)-3- hydroxy-2-(hydroxymethyl)propanoic acid (Compound 058)

[0482] Step one: synthesis of methyl 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)- 3-hydroxy-2-(hydroxymethyl)propanoate

[0483] Compound 1-5 (150.00 mg, 172.59 μmol), methyl 2-amino-3-hydroxy-2- (hydroxymethyl)propanoate (51.48 mg, 345.19 μmol), 2-(7-azobenzotriazole)-N,N,N',N'- tetramethyluronium hexafluorophosphate (131.26 mg, 345.19 μmol) and pyridine (27.30 mg, 345.19 μmol) were dissolved in N,N-dimethylformamide (1.5 mL), the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was quenched by adding water at 0 °C, then extracted with ethyl acetate (20.0 mL*3), the organic phase was washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound 28-1 (120.00 mg).

[0484] MS (ESI): m / z = 1000.40 [M+H] + .

[0485] Step two: synthesis of 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)- 3-hydroxy-2-(hydroxymethyl)propanoic acid

[0486] Lithium hydroxide (23.94 mg, 999.78 μmol) and compound 28-1 (100.00 mg, 99.98 μmol) were dissolved in a tetrahydrofuran (2.0 mL) and water (1.0 mL) solution, the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure, the pH value was adjusted to 6 with 2M HCl solution, concentrated under reduced pressure, the residue was purified by Prep-HPLC [YMC-TAR column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.05% formic acid) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 65%-95%] to give the target compound 058 (26.24 mg).

[0487] MS (ESI): m / z = 986.40 [M+H] + .

[0488] 1 H NMR (400 MHz, DMSO-d6) δ 12.28 (br s, 1H), 9.74 (s, 1H), 9.71 (s, 1H), 7.67 (s, 1H), 6.65 (s, 1H), 6.62 (s, 1H), 5.17 - 5.08 (m, 4H), 5.08 - 5.00 (m, 2H), 4.88 - 4.82 (m, 1H), 4.46 - 4.38 (m, 1H), 4.19 - 4.10 (m, 3H), 3.78 - 3.62 (m, 6H), 3.54 - 3.40 (m, 2H), 2.87 - 2.75 (m, 2H), 2.48 - 2.40 (m, 2H), 2.15 - 2.06 (m, 4H), 2.04 - 1.96 (m, 4H), 1.95 - 1.88 (m, 4H), 1.86 - 1.79 (m, 1H), 1.78 - 1.69 (m, 1H), 1.65 - 1.58 (m, 9H), 1.56 - 1.50 (m, 13H), 1.18 - 1.13 (m, 6H).

[0489] Example 29: Synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanamido)bicyclo[1.1.1]pentane-1-carboxylic acid (Compound 059)

[0490] Step one: Synthesis of methyl 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanamido)bicyclo[1.1.1]pentane-1-carboxylate

[0491] Compound 1-5 (100.00 mg, 115.06 μmol), methyl 3-aminobicyclo[1.1.1] pentane-1- carboxylate hydrochloride (55.15 mg, 310.51 μmol), 2-(7-azobenzotriazol)-N,N,N',N'- tetramethyluronium hexafluorophosphate (65.11 mg, 171.23 μmol) and N,N- diisopropylethylamine (44.61 mg, 345.19 μmol) were dissolved in N,N-dimethylformamide (10.0 mL), the reaction solution was stirred at room temperature for 2 hours. The reaction was quenched by adding water at 0 °C, then extracted with ethyl acetate (10.0 mL*3), the organic phase was washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the title compound 29-1 (90.00 mg).

[0492] Step two: synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido) bicyclo[1.1.1]pentane-1-carboxylic acid

[0493] Lithium hydroxide (21.72 mg, 907.04 μmol) and compound 29-1 (90.00 mg, 90.70 μmol) were dissolved in a tetrahydrofuran (2.0 mL) and water (1.0 mL) solution, the reaction solution was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction solution was concentrated under reduced pressure, the pH value was adjusted to pH = 6 with 2M HCl solution, concentrated under reduced pressure, and the residue was purified by Pre-HPLC [YMC-TAR column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.05% formic acid) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 75%-100%, elution time 9 minutes] to obtain the target compound 059 (38.48 mg).

[0494] MS (ESI): m / z = 978.4 [M+H] + .

[0495] 1H NMR (400 MHz, DMSO-d6) δ 9.79 (s, 2H), 8.69 (s, 1H), 6.65 (s, 1H), 6.63 (s, 1H), 5.24 - 5.08 (m, 4H), 5.08 - 4.99 (m, 2H), 4.71 - 4.60 (m, 1H), 4.43 - 4.35 (m, 1H), 4.21 - 4.10 (m, 3H), 3.79 - 3.68 (m, 2H), 3.52 - 3.44 (m, 2H), 2.89 - 2.74 (m, 2H), 2.48 - 2.41 (m, 2H), 2.16 - 2.11 (m, 2H), 2.09 - 2.05 (m, 2H), 2.04 - 1.96 (m, 4H), 1.96 - 1.88 (m, 4H), 1.80 - 1.71 (m, 2H), 1.64 - 1.56 (m, 10H), 1.55 - 1.50 (m, 12H), 1.49 - 1.39 (m, 2H), 1.20 - 1.12 (m, 6H).

[0496] Example 30: Synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)cyclobutane-1-carboxylic acid (Compound 060)

[0497] Step one: Synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)cyclobutane-1-carboxylic acid methyl ester

[0498] Compound 1-5 (100.00 mg, 115.06 μmol), 3-aminocyclobutane-1-carboxylic acid methyl ester hydrochloride (51.43 mg, 311.1 μmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65.11 mg, 171.23 μmol) and N,N-diisopropylethylamine (44.61 mg, 345.19 μmol) were dissolved in N,N-dimethylformamide (1.0 mL), the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was quenched by adding water at 0 °C, then extracted with ethyl acetate (10.0 mL*3), the organic phase was washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound 30-1 (90.00 mg).

[0499] MS (ESI): m / z = 980.50 [M+H] + .

[0500] Step two: synthesis of 3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)cyclobutane-1-carboxylic acid

[0501] Lithium hydroxide (21.99 mg, 918.15 μmol) and compound 30-1 (90.00 mg, 91.82 μmol) were dissolved in a tetrahydrofuran (2.0 mL) and water (1.0 mL) solution, the reaction solution was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction solution was concentrated under reduced pressure, the pH value was adjusted to pH = 6 with 2M HC1 solution, concentrated under reduced pressure, the residue was purified by Pre-HPLC [YMC-TAR column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.05% formic acid) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 85%-100%, elution time 10.3 minutes] to give the target compound 060 (17.24 mg).

[0502] MS (ESI): m / z = 966.50 [M+H] + .

[0503] 1 H NMR (400 MHz, DMSO-d6) δ 12.07 (s, 1H), 9.74 (s, 2H), 8.43 (d, J = 7.7 Hz, 1H), 6.64 (s, 1H), 6.62 (s, 1H), 5.17 - 5.08 (m, 4H), 5.08 - 4.99 (m, 2H), 4.70 - 4.63 (m, 1H), 4.38 (d, J = 17.0 Hz, 1H), 4.21 - 4.03 (m, 4H), 3.78 - 3.70 (m, 2H), 3.51 - 3.39 (m, 2H), 2.88 - 2.76 (m, 2H), 2.73 - 2.62 (m, 1H), 2.48 - 2.41 (m, 2H), 2.38 - 2.29 (m, 2H), 2.14 - 2.04 (m, 5H), 2.03 - 1.95 (m, 5H), 1.95 - 1.87 (m, 4H), 1.63 - 1.56 (m, 9H), 1.56 - 1.50 (m, 12H), 1.20 - 1.12 (m, 6H).

[0504] Example 31: Synthesis of (S)-3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoyl)thiazolidine-2-carboxylic acid (Compound 061)

[0505] Step one: Synthesis of (S)-thiazolidine-2-carboxylic acid

[0506] Compound 31-1 (500.00 mg, 2.14 mmol) was dissolved in 2M hydrochloric acid in dioxane (10 mL), and the reaction was stirred at 25 °C for 16 hours. LCMS showed the reaction was completed. The reaction was concentrated under reduced pressure to give the title compound 31-2 (280 mg).

[0507] 1 H NMR (400 MHz, DMSO-d6) δ 5.35 (s, 1H), 3.60-3.48 (m, 2H), 3.25-3.14 (m, 2H).

[0508] Step two: Synthesis of (S)-3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)thiazolidine-2-carboxylic acid

[0509] Compound 3-1 (330 mg, 341.55 μmol) and compound 31-2 (280 mg, 2.1 mmol) obtained from step one were dissolved in N,N-dimethylformamide (5 mL), and N,N- diisopropylethylamine (74.24 mg, 574.44 μmol) and N,N-dimethylaminopyridine (14 mg, 114.89 μmol) were added successively. The reaction was stirred at 25 °C for 16 hours. LCMS showed the reaction was completed. The reaction was purified by preparative high performance liquid chromatography (column: Boston Prime C18 150*30mm*5μm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 45%-65%) to give the title compound 061 (10.1 mg).

[0510] MS (ESI): m / z = 984.4 [M+H] +

[0511] 1H NMR (400 MHz, DMSO-d6) δ 9.97 - 9.63 (m, 2H), 6.71 - 6.58 (m, 2H), 5.22 - 4.88 (m, 8H), 4.31 - 4.06 (m, 4H), 4.04 - 3.86 (m, 1H), 3.78 - 3.68 (m, 2H), 3.56 - 3.46 (m, 1H), 3.11 - 2.92 (m, 2H), 2.87 - 2.77 (m, 2H), 2.49 - 2.40 (m, 2H), 2.24 - 1.81 (m, 13H), 1.80 - 1.32 (m, 25H), 1.21 - 1.12 (m, 6H).

[0512] Example 32: Synthesis of (R)-3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoyl)thiazolidine-2-carboxylic acid (Compound 062)

[0513] Step one: Synthesis of (R)-thiazolidine-2-carboxylic acid

[0514] Compound 32-1 (500.00 mg, 2.14 mmol) was dissolved in 2M hydrochloric acid in dioxane (10 mL), and the reaction was stirred at 25 °C for 16 hours. The reaction was complete according to LCMS. The reaction was concentrated under reduced pressure to give the title compound 32-2 (280 mg).

[0515] 1 H NMR (400 MHz, DMSO-d6) δ 5.35 (s, 1H), 3.61 - 3.49 (m, 2H), 3.30 - 3.05 (m, 2H).

[0516] Step two: Synthesis of (R)-3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoyl)thiazolidine-2-carboxylic acid

[0517] Compound 3-1 (330 mg, 341.55 pmol) and compound 32-2 (280 mg, 2.1 mmol) were dissolved in N,N-dimethylformamide (5 mL), N,N-diisopropylethylamine (74.24 mg, 574.44 pmol) and N,N-dimethylpyridine (14 mg, 114.89 pmol) were added. The reaction was stirred at 25 °C for 16 hours. LCMS showed the reaction was completed. The reaction was purified by preparative high performance liquid chromatography (column: Boston Prime C18 150*30mm*5pm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 43%-63%) to give the title compound 062 (16.2 mg).

[0518] MS (ESI): m / z = 984.3 [M+H] + .

[0519] 1 H NMR (400 MHz, DMSO-d6) d 10.03-9.52 (m, 2H), 6.69-6.61 (m, 2H), 5.32-4.93 (m, 8H), 4.26-4.06 (m, 4H), 3.99-3.90 (m, 1H), 3.76-3.70 (m, 2H), 3.54 - 3.42 (m, 1H), 3.15-3.08 (m, 1H), 2.99-2.92 (m, 1H), 2.87-2.74 (m, 2H), 2.49-2.41 (m, 2H), 2.22-1.84 (m, 13H), 1.81-1.73 (m, 2H), 1.66-1.42 (m, 21H), 1.20-1.12 (m, 6H).

[0520] Example 33: Synthesis of (S)-3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoyl)thiazolidine-4-carboxylic acid (Compound 063)

[0521] Compound 3-1 (166 mg, 171.81 pmol) and compound 33-1 (146.5 mg, 1.1 mmol) were dissolved in N,N-dimethylformamide (5 mL), N,N-diisopropylethylamine (111 mg, 859.07 pmol, 149.63 pL) and N,N-dimethylaminopyridine (21 mg, 171.81 pmol) were added. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 x 30 mm, 5 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 54% - 94%) to give the title compound 063 (9.1 mg).

[0522] MS (ESI): m / z = 984.3 [M+H] + .

[0523] 1 H NMR (400 MHz, DMSO-d6) d 9.85 (s, 1H), 9.76 (s, 1H), 6.74 - 6.58 (m, 2H), 5.29 - 5.20 (m, 1H), 5.19 - 5.08 (m, 4H), 5.08 - 5.02 (m, 2H), 4.84 - 4.74 (m, 1H), 4.54 - 4.23 (m, 1H), 4.22 - 4.07 (m, 4H), 3.78 - 3.69 (m, 2H), 3.54 - 3.46 (m, 2H), 3.23 - 2.99 (m, 2H), 2.90 - 2.72 (m, 2H), 2.49 - 2.41 (m, 2H), 2.17 - 1.70 (m, 14H), 1.69 - 1.34 (m, 21H), 1.20 - 1.11 (m, 6H).

[0524] Example 34: Synthesis of (R)-4-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoyl)morpholine-3-carboxylic acid (Compound 064)

[0525] Step one: Synthesis of (R)-4-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl) morpholine-3-carboxylic acid methyl ester Compound 1-5 (100.00 mg, 115.06 μmol), (R)-morpholine-3-carboxylic acid methyl ester (50.11 mg, 345.19 μmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65.11 mg, 171.23 μmol) and N,N-diisopropylethylamine (44.61 mg, 345.19 μmol) were dissolved in N,N-dimethylformamide (1.0 mL), the reaction solution was stirred at room temperature for 2 hours. The reaction was quenched by adding water at 0 °C, then extracted with ethyl acetate (10.0 mL*3), the organic phase was washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the title compound 34-1 (50.00 mg).

[0526] Step two: Synthesis of (R)-4-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl) morpholine-3-carboxylic acid

[0527] Lithium hydroxide (12 mg, 501.89 μmol) and compound 34-1 (50 mg, 50.19 μmol) were dissolved in a tetrahydrofuran (2.0 mL) and water (1.0 mL) solution, the reaction solution was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction solution was concentrated under reduced pressure, the pH value was adjusted to 6 with 2M HCl solution, concentrated under reduced pressure, and the residue was purified by Pre-HPLC [YMC-TAR column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 7 mmol / L NH4HCO3) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 35%-75%] to obtain the target compound 064 (13.46 mg).

[0528] MS (ESI): m / z = 982.4 [M+H] + .

[0529] 1H NMR (400 MHz, DMSO-d6) δ 6.74 - 6.69 (m, 1H), 6.69 - 6.66 (m, 1H), 5.35 - 5.20 (m, 1H), 5.16 - 4.93 (m, 5H), 4.76-4.66 (m, 1H), 4.44 - 4.23 (m, 2H), 4.18 (s, 2H), 4.14 - 4.01 (m, 2H), 3.81 - 3.63 (m, 4H), 3.60 - 3.46 (m, 3H), 3.44 - 3.30 (m, 2H), 3.16 - 3.05 (m, 2H), 2.99 - 2.76 (m, 3H), 2.53 - 2.44 (m, 2H), 2.14 - 2.04 (m, 4H), 2.03 - 1.85 (m, 9H), 1.62 - 1.52 (m, 13H), 1.50 - 1.44 (m, 10H), 1.25 - 1.13 (m, 8H).

[0530] Example 35: Synthesis of 1-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)- 4,4-difluoropyrrolidine-2-carboxylic acid (Compound 065)

[0531] Step one: Synthesis of 4,4-difluoropyrrolidine-2-carboxylic acid hydrochloride

[0532] Compound 35-1 (300 mg, 1.19 mmol) was dissolved in 2M hydrochloric acid in dioxane (5 mL), and the reaction was stirred at 25 °C for 16 hours. The reaction was complete according to LCMS. The reaction was concentrated under reduced pressure to give the title compound 35-2 (223.99 mg).

[0533] 1 H NMR (400 MHz, DMSO-d6) δ 4.67 (t, J = 8.5 Hz, 1H), 3.82 - 3.68 (m, 2H), 2.95 - 2.79 (m, 1H), 2.76 - 2.61 (m, 1H).

[0534] Step two: Synthesis of 1-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)- 4,4-difluoropyrrolidine-2-carboxylic acid

[0535] To a solution of compound 3-1 (166.75 mg, 172.59 pmol) in N,N- dimethylformamide (0.5 mL) was added N,N-diisopropylethylamine (89.22 mg, 690.36 pmol) and N,N-dimethylaminopyridine (21.08 mg, 172.59 pmol), followed by compound 35-2 (64.75 mg, 345.18 pmol). The reaction was stirred at 25 °C for 16 h. LCMS of the reaction showed the reaction was complete. The reaction was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 x 30 mm, 5 pm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 35% - 75%) to give the target compound 065 (14.6 mg).

[0536] MS (ESI): m / z = 1002.6 [M+H] + .

[0537] 1 H NMR (400 MHz, DMSO-d6) d 10.01 - 9.71 (m, 2H), 6.76 - 6.57 (m, 2H), 5.26 - 4.49 (m, 8H), 4.30 - 4.02 (m, 5H), 3.83 - 3.65 (m, 4H), 2.90 - 2.60 (m, 4H), 2.44 (br s, 2H), 2.22 - 1.83 (m, 13H), 1.77 - 1.66 (m, 2H), 1.64 - 1.40 (m, 25H), 1.20 - 1.12 (m, 6H).

[0538] Example 36: Synthesis of 2-(3,6-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)hexan-2- yl)propanedioic acid (Compound 066)

[0539] Step one: Synthesis of diethyl 2-(3,6-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)hexan-2- yl)propanedioate

[0540] Compound 36-2 (50 mg, 49.44 μmol) was dissolved in ethanol (0.8 mL) and water (0.2 mL), and lithium hydroxide (11.84 mg, 494.42 μmol) was added. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The ethanol was removed by concentration under reduced pressure. The mixture was adjusted to pH 5-6 with 1 N HC1 solution at 0 °C. Then extracted with ethyl acetate (20 mL*3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 27%-67%) to give the title compound 066 (3.9 mg).

[0541] MS (ESI): m / z = 1011.4 [M+H] + .

[0542] Step two: synthesis of 2-(3,6-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)hexan-2-yl)propanedioic acid

[0543] Compound 36-2 (50 mg, 49.44 μmol) was dissolved in ethanol (0.8 mL) and water (0.2 mL), and lithium hydroxide (11.84 mg, 494.42 μmol) was added. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The ethanol was removed by concentration under reduced pressure. The mixture was adjusted to pH 5-6 with 1 N HC1 solution at 0 °C. Then extracted with ethyl acetate (20 mL*3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 27%-67%) to give the title compound 066 (3.9 mg).

[0544] MS (ESI): m / z = 955.6 [M+H] + .

[0545] 1H NMR (400 MHz, DMSO-d6) δ 9.77-9.69 (m, 2H), 6.66-6.57 (m, 2H), 5.17-4.97 (m, 7H), 4.12-4.02 (m, 4H), 3.76-3.68 (m, 2H), 2.85-2.74 (m, 2H), 2.49-2.38 (m, 2H), 2.09 (s, 4H), 1.98 (s, 4H), 2.01-1.92 (m, 1H), 1.89 (s, 4H), 1.63-1.43 (m, 24H), 1.21-1.11 (m, 6H).

[0546] Example 37: Synthesis of ((((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentyl)oxy)carbonyl)glycine (Compound 067)

[0547] Step one: Synthesis of ((((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3-hydroxy-5-(methoxymethoxy)-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentyl)oxy)carbonyl)glycine

[0548] Compound 6-2 (200 mg, 212.04 μmol), glycine hydrochloride (47.30 mg, 424.08 μmol), di(p-nitrophenyl) carbonate (83.86 mg, 275.65 μmol) and N,N-diisopropyl ethylamine (109.62 mg, 848.17 μmol) were added into dichloroethane (2.00 mL). The reaction solution was stirred at 70 °C for 3 hours. LCMS showed that the reaction was completed. 30 mL of water was added to dilute the reaction solution, which was extracted with ethyl acetate (50 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude title compound 37-1 (300.00 mg).

[0549] MS (ESI): m / z = 1044.57 [M+H] + .

[0550] Step two: synthesis of (((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentyl)oxy) carbonyl)glycine

[0551] Compound 37-1 (300 mg, 287.28 pmol), pyridine p-toluenesulfonate (721.94 mg, 2.87 mmol) were added into tert-butanol (3.00 mL). The reaction was stirred at 80 °C for 16 h. LCMS showed the reaction was completed. The reaction was concentrated and purified by prep-HPLC [YMC TA-C18, 5 pm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 35-65%) as eluent] to give title compound 067 (8.50 mg).

[0552] MS (ESI): m / z = 956.5 [M+H] + .

[0553] 1 H NMR (400 MHz, DMSO-d6) d 9.79-9.72 (m, 2H), 6.85 (br s, 1H), 6.63 (s, 1H), 6.61 (s, 1H), 5.22-4.98 (m, 6H), 4.33 (s, 1H), 4.14 (d, J = 5.2 Hz, 5H), 3.77-3.70 (m, 2H), 3.43-3.25 (m, 4H), 2.86-2.75 (m, 2H), 2.49-2.39 (m, 2H), 2.16-1.84 (m, 14H), 1.64-1.41 (m, 24H), 1.20-1.12 (m, 6H).

[0554] Example 38: synthesis of (S)-2,5-bis((2R,3S)-6-cyclopropyl-2-((E)-4,8-dimethylnona-3,7- dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)- yl)pentanoic acid (Compound 068)

[0555] Cyclopropylboronic acid (117 mg, 1.36 mmol), compound 014 (200 mg, 194.76 μmol), di-tert-butyl (3-(tert-butyl)-4-methoxy-2,3-dihydrobenzo[d][1,3]oxaphosphinine-2- yl)phosphine oxide (14.96 mg, 38.95 μmol), potassium phosphate monobasic hydrate (135 mg, 584.29 μmol) and palladium(π-cinnamyl) chloride dimer (20 mg, 38.95 μmol) were added to toluene (3 mL) and the reaction was stirred at 100 °C under nitrogen atmosphere for 18 hours. LCMS showed the reaction was completed. The reaction was filtered and purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 80-90%) as eluents] to give title compound 068 (20 mg).

[0556] MS m / z (ESI): 949.4 [M+H] + .

[0557] 1 H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 8.18 (s, 1H), 5.19 - 5.07 (m, 4H), 5.08 - 4.99 (m, 2H), 4.72 - 4.63 (m, 1H), 4.20 (d, J = 16.8 Hz, 1H), 4.12 - 4.02 (m, 3H), 3.77 - 3.66 (m, 2H), 3.51 - 3.42 (m, 2H), 2.87 - 2.75 (m, 2H), 2.57 - 2.41 (m, 2H), 2.14 - 1.76 (m, 14H), 1.63 - 1.43 (m, 24H), 1.15 (s, 3H), 1.12 (s, 3H), 1.00 - 0.91 (m, 4H), 0.82 - 0.72 (m, 4H).

[0558] Example 39: Synthesis of (S)-3,6-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)hexanoic acid (Compound 069)

[0559] Step one: Synthesis of (2R, 2'R, 3S, 3'S)-((S)-5-chloro-5-oxopentane-l,4-diyl) bis(2- ((E)-4,8-dimethylnona-3,7-diene-l-yl)-2-methyl-7-oxo-2,3,4,7,8,9-hexahydropyrano[2,3- e]isoindol-8,3,5-triyl) tetraacetate

[0560] Compound 10-1 (440 mg, 424.20 μmol) was dissolved in dichloromethane (5 mL), 4 drops of N,N-dimethylformamide was added, then oxalyl chloride (107.69 mg, 848.49 μmol) was added dropwise at 0 °C. The reaction was stirred at 0 °C for 2 hours. The reaction was complete by LCMS (methanol quench). The reaction was concentrated under reduced pressure to give the title compound 39-1 (447.82 mg).

[0561] Step two: Synthesis of (2R, 2'R, 3S, 3'S)-((S)-6-diazo-5-oxohexane-l,4-diyl) bis(2- ((E)-4,8-dimethylnona-3,7-diene-l-yl)-2-methyl-7-oxo-2,3,4,7,8,9-hexahydropyrano[2,3- e]isoindol-8,3,5-triyl) tetraacetate

[0562] Compound 39-1 (447 mg, 423.42 μmol) was dissolved in tetrahydrofuran (4 mL) and acetonitrile (4 mL), (trimethylsilyl)diazomethane (2 M in ether, 529.28 μL) was added dropwise at 0 °C. The reaction was stirred at 25 °C for 16 hours. The reaction was complete by LCMS. The reaction was concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (gradient 0-30% tetrahydrofuran / petroleum ether, flow rate 35 mL / min) to give the title compound 39-2 (130 mg). 12g Flash column chromatography on silica gel (gradient 0-30% tetrahydrofuran / petroleum ether, flow rate 35 mL / min).

[0563] MS (ESI): m / z = 1033.6 [M-N2+H] + .

[0564] Step three: Synthesis of (2R, 2'R, 3S, 3'S)-((S)-6-methoxy-6-oxohexane-l,4-diyl) bis(2- ((E)-4,8-dimethylnona-3,7-diene-l-yl)-2-methyl-7-oxo-2,3,4,7,8,9-hexahydropyrano[2,3- e]isoindol-8,3,5-triyl) tetraacetate

[0565] Compound 39-2 (130 mg, 122.49 μmol) was dissolved in tetrahydrofuran (2 mL) and methanol (2 mL), silver nitrate (25.30 mg, 148.94 μmol) was added. The reaction was stirred at 25 °C for 16 h. LCMS of the reaction showed the reaction was complete. The reaction was concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography (silica gel, tetrahydrofuran / petroleum ether = 1 / 1, UV) to give the title compound 39-3 (20 mg).

[0566] MS (ESI): m / z = 1065.5 [M+H] + .

[0567] Step Four: Synthesis of (S)-3,6-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)hexanoic acid

[0568] Compound 39-3 (20 mg, 18.77 μmol) was dissolved in methanol (0.4 mL) and water (0.1 mL), lithium hydroxide monohydrate (15.77 mg, 375.48 μmol) was added. The reaction was stirred at 25 °C for 16 h. LCMS of the reaction showed the reaction was complete. The methanol was removed by concentrating the reaction under reduced pressure. The crude product was adjusted to pH 5-6 with 1 N HC1 solution at 0 °C. Then extracted with ethyl acetate (10 mL*3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 x 30 mm, 5 μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 38% - 78%) to give the title compound 069 (8.0 mg).

[0569] MS (ESI): m / z = 883.5 [M+H] + .

[0570] 1 H NMR (400 MHz, DMSO-d6) δ 9.79 - 9.73 (m, 2H), 6.64 - 6.58 (m, 2H), 5.25 - 4.93 (m, 6H), 4.48 - 4.38 (m, 1H), 4.16 - 4.09 (m, 4H), 3.76 - 3.68 (m, 2H), 2.89 - 2.78 (m, 2H), 2.65 - 2.55 (m, 2H), 2.45 - 2.40 (m, 1H), 2.17 - 1.85 (m, 13H), 1.79 - 1.35 (m, 25H), 1.20 - 1.12 (m, 6H).

[0571] Example 40: Synthesis of ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentyl) glycine (Compound 070)

[0572] Step one: Synthesis of ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentyl) methyl glycinate

[0573] Methyl 2-aminoacetate (31.33 mg, 351.66 μmol), sodium triacetoxyborohydride (62 mg, 293.05 μmol) and compound 7-2 (50 mg, 58.61 μmol) were added into N,N- dimethylformamide (1.00 mL). The reaction was stirred at room temperature for 1 hour. LCMS showed the reaction was completed. The reaction was cooled and poured into water (20 mL), extracted with ethyl acetate (30.00 mL*3), the organic layers were combined and washed with water (40.00 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give the title compound 40-1 (60 mg)

[0574] MS (ESI): m / z = 926.5 [M+H] + .

[0575] Step two: Synthesis of ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)pentyl) glycine

[0576] Compound 40-1 (60.00 mg, 64.78 μmol), lithium hydroxide monohydrate (5.44 mg, 129.56 μmol) were added into tetrahydrofuran (1.00 mL) and deionized water (1.00 mL). The reaction was stirred at room temperature for 1 hour. 2 mL 1M aqueous HC1 was added to adjust pH = 5, extracted with ethyl acetate (30 mL*3), the combined organic layers were washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, concentrated, purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent], lyophilized to give the title compound 070 (2 mg).

[0577] MS (ESI): m / z = 912.3 [M+H] + .

[0578] 1 H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1H), 9.75 (s, 1H), 6.66 (s, 1H), 6.61 (s, 1H), 5.21 - 4.99 (m, 6H), 4.45 - 4.40 (m, 1H), 4.26 - 3.97 (m, 4H), 3.75 - 3.70 (m, 2H), 3.23 - 3.01 (m, 4H), 2.92 - 2.75 (m, 4H), 2.49 - 2.41 (m, 2H), 2.22 - 1.85 (m, 14H), 1.65 - 1.43 (m, 24H), 1.20 - 1.06 (m, 6H).

[0579] Example 41: Synthesis of (S,E)-4,7-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)hept-2- enoic acid (Compound 071)

[0580] Step one: Synthesis of (S,E)-methyl 4,7-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1- yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-E]isoindol-8(2H)-yl)hept-2- enoate

[0581] Compound 7-2 (60.00 mg, 70.33 μmol), methoxyformylmethylene triphenylphosphonium (70.55 mg, 211.00 μmol) were added into dichloromethane (1.00 mL), the reaction was stirred at room temperature for 18 hours. The reaction was cooled and poured into water (20.00 mL), extracted with ethyl acetate (30.00 mL*3), the organic layers were combined and washed with water (40.00 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure to give the title compound 41-1 (60 mg)

[0582] MS m / z (ESI): 909.5 [M+H] + .

[0583] Step two: synthesis of (S, E)-4, 7-bis ((2R, 3S)-2-((E)-4, 8-dimethylnona-3, 7-dien-1-yl)-3, 5-dihydroxy-2-methyl-7-oxo-3, 4, 7, 9-tetrahydropyrano[2, 3-E] isoindol-8 (2H)-yl) hept-2-enoic acid

[0584] Compound 41-1 (50 mg, 55.00 μmol) and lithium hydroxide monohydrate (4.62 mg, 109.99 μmol) were added into tetrahydrofuran (3 mL) and deionized water (1 mL). The reaction was stirred at room temperature for 2 hours. 2 mL of 1M aqueous HC1 was added to the reaction to adjust pH = 5, extracted with ethyl acetate (30 mL*3), the organic layers were combined and washed with water (40 mL*2), saturated brine (40 mL), dried over anhydrous sodium sulfate. The drying agent was removed by filtration, purified by prep-HPLC [YMC TA-C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 60%-90%) as eluent], lyophilized to give the title compound 071 (2 mg).

[0585] MS (ESI): m / z = 895.4 [M+H] + .

[0586] 1H NMR (400 MHz, DMSO-d6) δ 9.86 - 9.80 (m, 2H), 8.40 (s, 1H), 6.66 (s, 1H), 6.62 (s, 1H), 5.66 - 5.58 (m, 1H), 5.22 - 4.98 (m, 6H), 4.84 - 4.76 (m, 1H), 4.20 - 3.95 (m, 4H), 3.76 - 3.70 (m, 2H), 3.36 - 3.20 (m, 2H), 2.95 - 2.63 (m, 2H), 2.54 - 2.47 (m, 2H), 2.20 - 1.72 (m, 14H), 1.74 - 1.43 (m, 24H), 1.18 (s, 3H), 1.15 (s, 3H).

[0587] Example 42: Synthesis of (S)-2,5-bis((2R,3S)-2-((4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2,6-dimethyl-7-oxo-3,4,7,9-tetrahydropyrano(2,3-e)isoindol-8(2H)-yl)pentanoic acid (Compound 072)

[0588] Compound 014 (80 mg, 77.9 μmol), trimethyl boroxine (29.34 mg, 233.72 μmol), potassium carbonate (32.30 mg, 233.72 μmol) were dissolved in dioxane (0.8 mL) / water (0.2 mL) under nitrogen protection, [(di(l-adamantyl)butylphosphino)-2-(2'-amino-1,1'- biphenyl)]palladium(II) methanesulfonate (11.35 mg, 15.58 μmol) was added, the reaction was stirred at 80 °C for 16 hours under nitrogen atmosphere. LCMS of the reaction showed the reaction was complete. The reaction was filtered and concentrated, the crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30mm, 5μm; mobile phase: [water (0.1% ammonia water)-acetonitrile]; gradient: 33%-73%), then by supercritical fluid chromatography (column: DAICEL CHIRALPAK IK (250mm*30mm, 10μm); mobile phase: [carbon dioxide-isopropanol (0.1% ammonia water)]; gradient: 50%-50%) to give the title compound 072 (2.8 mg).

[0589] MS (ESI): m / z = 897.3 [M+H] + .

[0590] 1H NMR (400 MHz, DMSO-d6) δ 8.54 - 8.46 (m, 2H), 5.17 - 5.09 (m, 4H), 5.07 - 5.02 (m, 2H), 4.72 - 4.62 (m, 1H), 4.27 - 4.16 (m, 1H), 4.15 - 4.03 (m, 3H), 3.75 - 3.69 (m, 2H), 3.48 - 3.43 (m, 2H), 2.93 - 2.81 (m, 2H), 2.43 - 2.35 (m, 6H), 2.09 (br d, J=4.5 Hz, 4H), 2.05 - 1.95 (m, 5H), 1.94 - 1.82 (m, 6H), 1.64 - 1.55 (m, 11H), 1.52 (s, 14H), 1.18 - 1.09 (m, 6H).

[0591] Example 43: Synthesis of (S)-2,5-bis((2R,3S)-6-cyano-2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9- tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoic acid (Compound 073)

[0592] Compound 014 (100 mg, 97.38 μmol), potassium ferrocyanide trihydrate (246.80 mg, 584.29 μmol) were dissolved in dioxane (0.6 mL) and water (0.2 mL) under nitrogen protection, and then (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'- biphenyl) palladium(II) (77.36 mg, 97.38 μmol), allylpalladium(II) chloride dimer (35.63 mg, 97.38 μmol) were added. The reaction was stirred at 100 °C for 16 hours. LCMS showed the reaction was complete. The reaction was filtered and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 x 30 mm, 5 μm; mobile phase: [water (0.05% ammonia water-0.001% ammonium bicarbonate)-acetonitrile]; gradient: 12%-52%) to give the title compound 073 (12.3 mg).

[0593] MS (ESI): m / z = 919.5 [M+H] + .

[0594] 1H NMR (400 MHz, DMSO-d6) δ 5.41 - 5.15 (m, 1H), 5.13 - 4.96 (m, 4H), 4.68 - 4.59 (m, 1H), 4.35 - 4.12 (m, 4H), 3.81 - 3.74 (m, 2H), 3.54 - 3.46 (m, 2H), 2.89 - 2.79 (m, 2H), 2.60 - 2.51 (m, 2H), 2.16 - 1.81 (m, 15H), 1.63 - 1.47 (m, 24H), 1.28 - 1.15 (m, 6H).

[0595] Example 44: Synthesis of 4-((S)-1,4-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)butyl)thiazole- 2-carboxylic acid (Compound 081)

[0596] Step one: Synthesis of (2R,2'R,3S,3'S)-((S)-6-chloro-5-oxohexane-1,4-diyl) bis(2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-2-methyl-7-oxo-2,3,4,7,8,9-hexahydropyrano[2,3-e]isoindol-8,3,5- triyl) tetraacetate

[0597] Compound 39-1 (447 mg, 423.42 μmol) was dissolved in tetrahydrofuran (4 mL) and acetonitrile (4 mL), (trimethylsilyl)diazomethane (2 M in ether, 529 μL) was added dropwise at 0 °C. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction was concentrated under reduced pressure, the crude product was purified by flash silica gel column chromatography (gradient 0-30% tetrahydrofuran / petroleum ether, flow rate 35 mL / min) to give the title compound 44-1 (230 mg). 12g Flash silica gel column, gradient 0-30% tetrahydrofuran / petroleum ether, flow rate 35 mL / min) to give the title compound 44-1 (230 mg).

[0598] Step two: Synthesis of 4-((S)-1,4-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- diacetoxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)butyl)thiazole-2- carboxylic acid ethyl ester

[0599] Compound 44-1 (230 mg, 215.01 μmol) was dissolved in ethanol (3 mL), and 2-amino-2-thioacetic acid ethyl ester (42.95 mg, 322.52 μmol) was added. The reaction was stirred at 80 °C for 16 h. The reaction was concentrated under reduced pressure, and the residue was added to pyridine (3 mL) and acetic anhydride (219.50 mg, 2.15 mmol). The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction was concentrated under reduced pressure. The crude product was purified by preparative thin layer chromatography (silica, petroleum ether: tetrahydrofuran = 1:1) to give the title compound 44-2 (150 mg).

[0600] MS (ESI): m / z = 1148.4 [M+H] + .

[0601] Step Three: Synthesis of 4-((S)-1,4-bis((2R,3S)-2-((E)-4,8-dimethylnon-3,7-dien-1 -yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)butyl)thiazole- 2-carboxylic acid

[0602] Compound 44-2 (120 mg, 104.49 μmol) was dissolved in methanol (2 mL) and water (0.5 mL), and lithium hydroxide monohydrate (50.05 mg, 1.19 mmol) was added. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The methanol was removed by concentration under reduced pressure. The mixture was adjusted to pH 5-6 with 1 N HC1 at 0 °C. Then extracted with ethyl acetate (20 mL*3). The organic phase was dried and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 34%-74%) to give the target compound 081 (2.0 mg).

[0603] MS (ESI): m / z = 952.5 [M+H] + .

[0604] 1H NMR (400 MHz, DMSO-d6) δ 9.81 - 9.73 (m, 2H), 7.25 - 7.07 (m, 1H), 6.68 - 6.59 (m, 2H), 5.55 - 5.36 (m, 1H), 5.20 - 4.95 (m, 6H), 4.30 - 4.09 (m, 3H), 3.96 - 3.89 (m, 1H), 3.74 - 3.69 (m, 2H), 2.88 - 2.74 (m, 3H), 2.46 - 2.38 (m, 2H), 2.15 - 1.81 (m, 15H), 1.64 - 1.39 (m, 24H), 1.21 - 1.12 (m, 6H).

[0605] Example 45: Synthesis of N-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)- N-(2-oxo-2-(((1R,2S,4R)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-yl)oxy)ethyl)glycine (Compound 082)

[0606] Step one: Synthesis of N-Boc glycine dextran ester

[0607] N-Boc glycine (2.00 g, 11.42 mmol), dextran (1.94 g, 12.56 mmol) were dissolved in dichloromethane (25 mL), N,N'-dicyclohexylcarbodiimide (2.36 g, 11.42 mmol) and 4-dimethylaminopyridine (139.48 mg, 1.14 mmol) were added slowly into the reaction solution. The reaction solution was stirred at room temperature for 4 hours. LCMS showed the reaction was complete. The reaction solution was filtered, the filtrate was collected and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound 45-2 (3.56 g) which was used directly in the next step without further purification.

[0608] MS (ESI): m / z = 212.20 [M+H-100] + .

[0609] Step two: Synthesis of glycine dextran ester hydrochloride

[0610] N-Boc glycine dextran ester (3.00 g, 9.63 mmol) was dissolved in hydrogen chloride / 1,4-dioxane solution (4 M, 12 mL, 48 mmol). The reaction solution was stirred at room temperature for 2 hours. The solvent in the reaction solution was removed by concentration under reduced pressure to give the title compound 45-3 (1.00 g) as a white solid.

[0611] 1 H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 3H), 4.94-4.87 (m, 1H), 3.79 (s, 2H), 2.35-2.23 (m, 1H), 1.98-1.87 (m, 1H), 1.76-1.66 (m, 2H), 1.34-1.17 (m, 2H), 1.10-0.98 (m, 1H), 0.88 (s, 3H), 0.85 (s, 3H), 0.82 (s, 3H).

[0612] Step three: synthesis of ethyl (2-oxo-2-(((1R,2S,4R)-1,7,7-trimethylbicyclo[2.2.1]heptan-2- yl)oxy)ethyl)glycinate

[0613] Glycine nortryptophan hydrochloride (1.00 g, 4.04 mmol) was dissolved in N,N- dimethylformamide (10 mL), the reaction bottle was placed in an ice water bath, triethylamine (408.43 mg, 4.04 mmol) was added, followed by dropwise addition of ethyl bromoacetate (338 mg, 2.02 mmol). The reaction was reacted at 0 °C for 1 hour. LCMS showed that the reaction was complete. 10 mL of water was added to quench, extracted with ethyl acetate (10 mL*3), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography (silica, ethyl acetate / petroleum ether = 10%-50%) to give the title compound 45-4 (220 mg).

[0614] MS (ESI): m / z = 298.20 [M+H] + .

[0615] Step four: synthesis of N-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)- N-(2-oxo-2-(((1R,2S,4R)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-yl)oxy)ethyl)glycine ethyl ester

[0616] Compound 1-5 (110 mg, 126.57 μmol) and compound 45-4 (75.28 mg, 253.14 μmol) were dissolved in N,N-dimethylformamide (2 mL), N,N- diisopropylethylamine (49 mg, 379.11 μmol) and 2-(7-azabenzotriazol-1- yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (57 mg, 149.90 μmol) were added. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction was added ethyl acetate (30 mL), then washed with saturated brine (10 mL*4). The organic phase was dried and concentrated to give the title compound 45-5 (145 mg).

[0617] Step five: synthesis of N-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)- N-(2-oxo-2-(((1R,2S,4R)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-yl)oxy)ethyl)glycine

[0618] Compound 45-5 (145 mg, 126.26 μmol) was dissolved in tetrahydrofuran (3 mL), lithium hydroxide aqueous solution (0.5 M, 758 μL) was added. The reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was completed. The reaction was adjusted pH = 5-6 with 1 N hydrochloric acid aqueous solution, then extracted with ethyl acetate (30 mL*2), the organic phase was combined and concentrated under reduced pressure, the residue was prepared by high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 42%-82%) to give the target compound 082 (22.7 mg).

[0619] MS (ESI): m / z = 1120.5 [M+H] + .

[0620] 1H NMR (400 MHz, DMSO-d6) δ 9.91 - 9.65 (m, 2H), 6.70 - 6.56 (m, 2H), 5.22 - 4.98 (m, 7H), 4.88 - 4.58 (m, 2H), 4.41 - 3.90 (m, 8H), 3.78 - 3.67 (m, 2H), 2.87 - 2.75 (m, 2H), 2.36 - 2.21 (m, 1H), 2.15 - 1.99 (m, 8H), 1.95 - 1.88 (m, 4H), 1.79 - 1.70 (m, 2H), 1.63 - 1.53 (m, 21H), 1.46 - 1.38 (m, 2H), 1.23 (s, 1H), 1.21 - 1.13 (m, 8H), 0.98 - 0.90 (m, 1H), 0.89 - 0.81 (m, 4H), 0.74 - 0.63 (m, 6H), 0.73 - 0.68 (m, 3H), 0.64 (s, 2H).

[0621] Example 46: Synthesis of 5-((S)-1,4-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)butyl)oxazole- 4-carboxylic acid (Compound 083)

[0622] Step one: Synthesis of 2,5-dioxopyrrolidin-1-yl (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7- dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoate

[0623] Compound 1-5 (200 mg, 230.13 μmol), N-hydroxysuccinimide (31.78 mg, 276.15 μmol) were dissolved in N,N dimethylformamide (3 mL), dicyclohexyl carbodiimide (56.98 mg, 276.15 μmol) was added, the reaction was stirred at 25 °C for 16 hours. LCMS showed the reaction was completed, the title compound 3-1 was obtained. The reaction was directly used for the next step.

[0624] Step two: Synthesis of 5-((S)-1,4-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy- 2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)butyl)oxazole-4-carboxylic acid ethyl ester

[0625] Compound 3-1 (222 mg, 229.77 pmol) was dissolved in N,N-dimethylformamide (3 mL), ethyl isocyanacetate (39.04 mg, 345.14 pmol) and cuprous oxide (16.46 mg, 115.04 pmol) were added. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction was added into ethyl acetate (100 mL), then washed with saturated brine (20 mL*4). The organic phase was dried and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 pm; mobile phase: [water (0.05% ammonia water + 0.001% ammonium bicarbonate) - acetonitrile]; gradient: 53% - 100%) to give the title compound 46-1 (20 mg).

[0626] MS (ESI): m / z = 964.2 [M+H] + .

[0627] Step three: synthesis of 5-((S)-1,4-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)butyl)oxazole-4-carboxylic acid

[0628] Compound 46-1 (20 mg, 20.74 pmol) was dissolved in tetrahydrofuran (0.6 mL), lithium hydroxide aqueous solution (0.5 M, 125 pL) was added. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction was adjusted to pH = 5-6 with 1 N HC1, then extracted with ethyl acetate (10 mL*3). The organic phase was dried and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 59% - 99%) to give the target compound 083 (2.1 mg).

[0629] MS (ESI): m / z = 936.2 [M+H] + .

[0630] 1H NMR (400 MHz, DMSO-d6) d = 9.93-9.62 (m, 2H), 8.34 (s, 1H), 6.65-6.58 (m, 2H), 6.13-6.05 (m, 1H), 5.19-4.95 (m, 6H), 4.45-4.36 (m, 1H), 4.24-4.10 (m, 3H), 3.78-3.71 (m, 2H), 2.85-2.75 (m, 2H), 2.46-2.39 (m, 1H), 2.13-1.83 (m, 15H), 1.72-1.40 (m, 26H), 1.21-1.10 (m, 6H).

[0631] Example 47: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-6-(trifluoromethyl)-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanoic acid (Compound 084)

[0632] Step one: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-6-iodo-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)- yl)pentanoic acid

[0633] Compound 1-5 (1 g, 1.15 mmol) was added to tetrahydrofuran (10 mL), and N-iodosuccinimide (724.83 mg, 3.22 mmol) was added at -20 °C. The reaction solution was stirred at room temperature for 6 h. The reaction solution was quenched by adding 20 mL of saturated aqueous sodium sulfite solution, extracted with dichloromethane (30 mL*2), the combined organic layers were washed with water (40 mL*2), saturated brine (40 mL), and dried over anhydrous sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica, dichloromethane / methanol = 10 / 1) to give the title compound 47-1 (700 mg).

[0634] Step two: Synthesis of methoxymethyl (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7- dien-1-yl)-3-hydroxy-6-iodo-5-(methoxymethoxy)-2-methyl-7-oxo-3,4,7,9- tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoate

[0635] Compound 47-1 (0.5 g, 446.07 μmol) was dissolved in dichloromethane (10 mL) under nitrogen atmosphere, N,N-diisopropylethylamine (576.51 mg, 4.46 mmol) and bromomethyl methyl ether (359.49 mg, 2.88 mmol) were added. The reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was complete. The reaction was concentrated under reduced pressure, the crude product was purified by flash silica gel column chromatography (0 ~ 50% petroleum ether / tetrahydrofuran, flow rate 30 mL / min) to give the title compound 47-2 (246 mg). 12g Flash silica gel column, gradient 0 ~ 50% petroleum ether / tetrahydrofuran, flow rate 30 mL / min) to give the title compound 47-2 (246 mg).

[0636] MS (ESI): m / z = 1253.2 [M+H] + .

[0637] Step three: Synthesis of methoxymethyl (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3-hydroxy-6-(trifluoromethyl)-5-(methoxymethoxy)-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoate

[0638] In the glove box, cuprous chloride (79.01 mg, 798.08 μmol) and potassium tert-butoxide (179.10 mg, 1.60 mmol) were added to N,N dimethylformamide (5 mL), stirred at room temperature for 30 min, phenyl trifluoromethylsulfoxide (216.94 mg, 1.12 mmol) was added, stirred at room temperature for 30 min. After the pre-generated trifluoromethyl cuprous stabilized by triethylamine hydrofluoric acid salt (128.66 mg, 798.06 μmol), compound 47-2 (200 mg, 159.61 μmol) was added. The reaction was stirred at 80 °C for 16 h. LCMS showed the reaction was complete. Diluted with ethyl acetate (30 mL), washed with saturated brine (20 mL*3), dried and concentrated, the crude product was purified by preparative thin layer chromatography (silica, petroleum ether: tetrahydrofuran = 1:1) to give the title compound 47-3 (36 mg).

[0639] MS (ESI): m / z = 1137.5 [M+H] + .

[0640] Step four: Synthesis of (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-6-(trifluoromethyl)-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoic acid

[0641] Compound 47-3 (35 mg, 30.78 pmol) was dissolved in isopropanol (1 mL), pyridine p-toluenesulfonate (116.01 mg, 461.64 pmol) was added. The reaction was stirred at 80 °C for 16 h. LCMS showed the reaction was complete. The reaction was concentrated, diluted with ethyl acetate (20 mL), washed with saturated brine (10 mL*3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 62% - 100%) to give compound 084 (1.1 mg).

[0642] MS (ESI): m / z = 1005.5 [M+H] + .

[0643] 1 H NMR (400 MHz, DMSO-d6) d 9.86 - 8.74 (m, 1H), 5.36 - 5.27 (m, 2H), 5.20 - 4.97 (m, 4H), 4.77 - 4.60 (m, 1H), 4.37 - 4.07 (m, 4H), 3.87 - 3.71 (m, 2H), 3.56 - 3.44 (m, 2H), 2.92 - 2.81 (m, 2H), 2.65 - 2.55 (m, 2H), 2.17 - 1.80 (m, 15H), 1.69 - 1.42 (m, 24H), 1.26 - 1.15 (m, 6H).

[0644] Example 48: Synthesis of (3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanamido)propanoyl)-L-histidine (Compound 085)

[0645] Step one: Synthesis of (3-aminopropanoyl)-L-histidine methyl ester

[0646] The starting material (3-aminopropanoyl)-L-histidine (1 g, 4.42 mmol) was dissolved in methanol (10 mL), dichlorosulfoxide (788.81 mg, 6.63 mmol) was added dropwise slowly at 0 °C, the reaction was stirred at 60 °C for 16 h. LCMS showed the reaction was complete. The reaction was concentrated to give the title compound 48-2 (1.06 g) which was used directly in the next step.

[0647] Step 2: Synthesis of (S)-tert-butyl 4-(2-(3-((tert-butoxycarbonyl)amino) propanamido)-3-methoxy-3-oxopropyl)-lH-imidazole-l-carboxylate

[0648] Compound 48-2 (1 g, 4.16 mmol), di-tert-butyl dicarbonate (1.82 g, 8.34 mmol) were dissolved in dichloromethane (20 mL), triethylamine (1.68 g, 16.65 mmol) was added, and the reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction was concentrated, and the crude product was purified by flash silica gel column chromatography (gradient 0-30% petroleum ether / tetrahydrofuran at a flow rate of 30 mL / min) to give the title compound 48-3 (795 mg). 12g Flash silica gel column, gradient 0-30% petroleum ether / tetrahydrofuran at a flow rate of 30 mL / min) to give the title compound 48-3 (795 mg).

[0649] MS (ESI): m / z = 441.1 [M+H] + .

[0650] Step 3: Synthesis of (3-aminopropanoyl)-L-histidine methyl ester hydrochloride

[0651] Compound 48-3 (750 mg, 1.70 mmol) was dissolved in 2M hydrochloric acid in dioxane (20 mL), and the reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction was concentrated to give the title compound 48-4 (471 mg).

[0652] Step 4: Synthesis of (3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido) propanoyl)-L-histidine methyl ester

[0653] Compound 1-5 (200 mg, 230.12 μmol), compound 48-4 (127.36 mg, 460.25 μmol) were dissolved in N,N dimethylformamide (5 mL), diisopropylethylamine (118.97 mg, 920.50 μmol) and 2-(7-azabenzotriazol-1- yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (104.18 mg, 273.98 μmol) were added, and the reaction was stirred at room temperature for 16 h. LCMS showed the reaction was complete. The reaction was diluted with ethyl acetate (30 mL), washed with saturated brine (20 mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the title compound 48-5 (251 mg).

[0654] Step five: synthesis of (3-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido) propanoyl)-L-histidine

[0655] Compound 48-5 (250 mg, 229.07 μmol) was dissolved in tetrahydrofuran (4 mL) and water (1 mL), and lithium hydroxide monohydrate (48.06 mg, 1.14 mmol) was added. The reaction was stirred at 25 °C for 16 hours. LCMS showed the reaction was complete. The pH of the reaction was adjusted to 5-6 with 1 N HC1 at 0 °C. Then extracted with ethyl acetate (20 mL*3). The organic phase was dried and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [water (0.05% ammonia water + 10 mM ammonium bicarbonate) - acetonitrile]; gradient: 31%-71%) to give the target compound 085 (61 mg).

[0656] MS (ESI): m / z = 1077.7 [M+H] + .

[0657] 1 H NMR (400 MHz, DMSO-d6) d = 9.93-9.60 (m, 1H), 8.25-7.99 (m, 2H), 7.56 (s, 1H), 6.78 (s, 1H), 6.67-6.60 (m, 2H), 5.16-4.99 (m, 5H), 4.74-4.68 (m, 1H), 4.43-4.33 (m, 2H), 4.22-4.12 (m, 3H), 3.77-3.70 (m, 3H), 3.51-3.41 (m, 3H), 3.23-3.14 (m, 2H), 2.95-2.78 (m, 4H), 2.47-2.41 (m, 1H), 2.30-2.21 (m, 2H), 2.17-1.85 (m, 14H), 1.83-1.73 (m, 2H), 1.64-1.57 (m, 10H), 1.56-1.50 (m, 13H), 1.20-1.13 (m, 6H).

[0658] Example 49: Synthesis of (S)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanamido)-3-(2-mercapto-1H-imidazol-4-yl)propanoic acid (Compound 086)

[0659] Step one: Synthesis of (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(2-thioxo-2,3- dihydro-1H-imidazol-4-yl)propanoate

[0660] Compound 49-1 (1 g, 3.71 mmol) was dissolved in dioxane (14.52 mL) under nitrogen protection, and phenyl chlorothionoformate (3.21 g, 18.59 mmol) was added dropwise at 0 °C. The reaction was stirred at 25 °C for 16 h. Water (11.13 mL) was added, and the reaction was stirred at 25 °C for another 16 h. The reaction was extracted with ethyl acetate (50 mL*2). The organic phase was dried and concentrated. The crude product was added with methanol (21.30 mL) and triethylamine (1.13 g, 11.17 mmol), and stirred at 25 °C for another 16 h. LCMS showed the reaction was complete. The reaction was concentrated under reduced pressure. Ethyl acetate (50 mL) was added, filtered, and the filtrate was concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [water (0.05% ammonia water + 10 mM ammonium bicarbonate) - acetonitrile]; gradient: 4%-44%) to give the title compound 49-2 (0.24 g).

[0661] MS (ESI): m / z = 302.1 [M+H] + .

[0662] Step two: Synthesis of (S)-methyl 2-amino-3-(2-thioxo-2,3-dihydro-1H-imidazol-4- yl)propanoate hydrochloride

[0663] Compound 49-2 (190 mg, 630.47 μmol) was dissolved in 2M hydrochloric acid dioxane (5 mL), and the reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction was concentrated to give the title compound 49-3 (149 mg).

[0664] 1 H NMR (400 MHz, DMSO-d6) δ 8.76 (br s, 3H), 6.89 (br s, 1H), 4.29 (br s, 1H), 3.72 (s, 3H), 3.12-2.97 (m, 2H).

[0665] Step three: Synthesis of (S)-methyl 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8- dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2H)-yl)pentanamido)-3-(2-mercapto-1H-imidazol-4-yl)propanoate

[0666] Compound 1-5 (200 mg, 230.12 umol), compound 49-3 (82.05 mg, 345.19 umol) were dissolved in N,N dimethylformamide (1.41 mL), N,N-diisopropylethylamine (118.97 mg, 920.46 umol) and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (104.18 mg, 273.98 umol) were added. The reaction was stirred at room temperature for 16 hours. LCMS showed the reaction was completed. The reaction was diluted with ethyl acetate (30 mL) and washed with saturated brine (20 mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30mm, 5um; mobile phase: [water (0.05% ammonia water + 10mM ammonium bicarbonate) - acetonitrile]; gradient: 59%-99%) to give the title compound 49-4 (40 mg).

[0667] MS (ESI): m / z = 1052.6 [M+H] + .

[0668] Step four: Synthesis of (S)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)-3- (2-mercapto-1H-imidazol-4-yl)propanoic acid

[0669] Compound 49-4 (40 mg, 38.01 μmol) was dissolved in tetrahydrofuran (1 mL) and water (0.25 mL), and lithium hydroxide monohydrate (7.98 mg, 190.18 μmol) was added. The reaction was stirred at 25 °C for 16 h. LCMS of the reaction showed it was complete. The pH of the reaction was adjusted to 5-6 with 1 N HC1 at 0 °C. It was then extracted with ethyl acetate (20 mL*3). The organic phase was dried and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [water (0.05% ammonia water + 10 mM ammonium bicarbonate) - acetonitrile]; gradient: 40%-60%) to give the target compound 086 (7.5 mg).

[0670] MS (ESI): m / z = 1038.7 [M+H] + .

[0671] 1 H NMR (400 MHz, DMSO-d6) d = 11.80-11.29 (m, 1H), 10.03-9.56 (m, 2H), 8.22-7.93 (m, 1H), 6.68-6.60 (m, 2H), 6.45 (s, 1H), 5.26-4.99 (m, 6H), 4.83-4.74 (m, 1H), 4.46-4.37 (m, 1H), 4.26-3.97 (m, 4H), 3.80-3.71 (m, 2H), 3.59-3.44 (m, 4H), 2.90-2.78 (m, 2H), 2.73-2.66 (m, 2H), 2.46-2.40 (m, 1H), 2.20-1.67 (m, 15H), 1.66-1.44 (m, 24H), 1.20-1.13 (m, 6H).

[0672] Example 50: Synthesis of (S)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)- 4,4,4-trifluorobutanoic acid and (R)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)- 4,4,4-trifluorobutanoic acid (Compounds 087 & 088)

[0673] Step 1: Synthesis of methyl 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)- 4,4,4-trifluorobutanoate

[0674] Methyl 2-amino-4,4,4-trifluorobutanoate (78.76 mg, 460.25 pmol) and compound 1-5 (200 mg, 230.13 pmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (131.25 mg, 345.19 pmol), N,N-diisopropylethylamine (118.97 mg, 920.50 pmol) were added into N,N-dimethylformamide (3 mL). The reaction was stirred at room temperature for 3 h. LCMS showed the reaction was completed. The reaction was filtered, the filtrate was purified by prep-HPLC [YMC TA-C18, 5 pm silica, 30 mm diameter, 150 mm length; using a mixture of water (with 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 80-90%) as eluent] and lyophilized to give the title compound 50-1 (100 mg).

[0675] Step 2: Synthesis of (S)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)-4,4,4- trifluorobutanoic acid and (R)-2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5- dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)-4,4,4- trifluorobutanoic acid

[0676] Compound 50-1 (100 mg, 97.83 pmol) and lithium hydroxide monohydrate (41.09 mg, 978.29 pmol) were added into tetrahydrofuran (2 mL) and water (1 mL). The reaction was stirred at room temperature for 2 h. LCMS showed the reaction was completed. The reaction was filtered, the filtrate was purified by prep-HPLC [YMC TA-C18, 5 pm silica, 30 mm diameter, 150 mm length; using a mixture of water (with 7 mmol / L ammonium bicarbonate) and acetonitrile (acetonitrile content: 89-92%) as eluent] and lyophilized to give the target compound 087 (10 mg) and compound 088 (8 mg).

[0677] Compound 087:

[0678] HPLC [Column: Agilent Poroshell 120 EC-C18 (4,6*100, 2.7 pm); Mobile phase: 0.1% formic acid-acetonitrile / 0.1% formic acid-water, gradient (acetonitrile gradient ratio 10%-95%) elution for 18 minutes, isocratic (acetonitrile gradient ratio 10%) elution for 5 minutes; flow rate 1.0 mL / min], Retention time: 13.004 min.

[0679] MS m / z (ESI): 1008.3 [M+H] + .

[0680] 1 H NMR (400 MHz, DMSO-d6) δ 9.82 - 9.74 (m, 2H), 6.66 (s, 1H), 6.62 (s, 1H), 5.18 - 4.99 (m, 6H), 4.80 - 4.71 (m, 1H), 4.31 - 4.22 (m, 1H), 4.20 - 4.04 (m, 4H), 3.77 - 3.69 (m, 2H), 3.54 - 3.32 (m, 2H), 2.86 - 2.63 (m, 4H), 2.50 - 2.39 (m, 4H), 2.22 - 1.80 (m, 14H), 1.68 - 1.39 (m, 24H), 1.17 (s, 3H), 1.14 (s, 3H).

[0681] Compound 088:

[0682] HPLC [Column: Agilent Poroshell 120 EC-C18 (4,6*100, 2.7 pm); Mobile phase: 0.1% formic acid-acetonitrile / 0.1% formic acid-water, gradient (acetonitrile gradient ratio 10%-95%) elution for 18 minutes, isocratic (acetonitrile gradient ratio 10%) elution for 5 minutes; flow rate 1.0 mL / min], Retention time: 13.619 min.

[0683] MS m / z (ESI): 1008.4 [M+H] + .

[0684] 1H NMR (400 MHz, DMSO-d6) δ 9.81 - 9.73 (m, 2H), 6.65 (s, 1H), 6.61 (s, 1H), 5.19 - 5.01 (m, 6H), 4.84 - 4.75 (m, 1H), 4.47 - 4.38 (m, 1H), 4.17 - 4.08 (m, 4H), 3.77 - 3.69 (m, 2H), 3.55 - 3.28 (m, 2H), 2.86 - 2.59 (m, 4H), 2.50 - 2.39 (m, 4H), 2.23 - 1.79 (m, 14H), 1.75 - 1.39 (m, 24H), 1.16 (s, 3H), 1.14 (s, 3H).

[0685] Example 51: Synthesis of ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)- D-alanine (Compound 023)

[0686] Method 1:

[0687] Step 1: Synthesis of methyl ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)- D-alaninate

[0688] Compound 1-5 (600 mg, 690.36 umol), D-alanine methyl ester (142.38 mg, 1.38 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (524.72 mg, 1.38 mmol) and N,N-diisopropylethylamine (267.67 mg, 2.07 mmol) were dissolved in N,N-dimethylformamide (10.0 mL), the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was quenched by adding water at 0 °C, then extracted with ethyl acetate (50*3 mL), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound 51-1 (500 mg).

[0689] MS (ESI): m / z = 954.6 [M+H]+.

[0690] Step 2: Synthesis of ((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoyl)-D-alanine

[0691] Lithium hydroxide (125.50 mg, 5.24 mmol) and compound 51-1 (500.00 mg, 524.00 μmol) were dissolved in tetrahydrofuran (8 mL) and water (4 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored for completion by LCMS. The reaction mixture was concentrated under reduced pressure, the pH adjusted to pH 6 with 2M HCl, and then concentrated under reduced pressure. The residue was purified by prep-HPLC using a YMC-TAR column (5 μm silica, 30 mm diameter, 150 mm length); eluents were a decreasingly polar mixture of water (containing 7 mmol / L NH4HCO3) and acetonitrile; the acetonitrile gradient was 35% to 65% over 10 minutes) to yield the title compound 023 (175 mg).

[0692] MS (ESI): m / z = 940.4 [M+H] + .

[0693] 1 H NMR (400MHz, DMSO-d6) δ12.46(brs,1H),9.80-9.66(m,2H),8.38(d,J=7.2Hz,1H),6.65(s,1H),6.62(s,1H),5. 18–5.08(m,4H),5.08–5.00(m,2H),4.89–4.80(m,1H),4.44(d,J=17.1Hz,1H),4.21–4.08(m,4H),3.78–3.67(m, 2H),3.47(t,J=6.9Hz,2H),2.88–2.77(m,2H),2.47–2.39(m,2H),2.10(d,J=8.8Hz,4H),2.05–1.95(m,4H),1.9 5–1.86(m,4H),1.84–1.68(m,2H),1.64–1.57(m,9H),1.56–1.51(m,12H),1.28-1.20(m,4H),1.20-1.12(m,6H).

[0694] Method 2:

[0695] by Compound 023 was prepared by replacing compound 1-1 with reference to Example 1 as the raw material. MS (ESI): m / z = 940.3 [M+H]+ .

[0696] Example 52: Synthesis of 1-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)cyclopropane-1-carboxylic acid (Compound 030)

[0697] Step one: Synthesis of 2,5-dioxopyrrolidin-1-yl (S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanoate

[0698] Compound 1-5 (100 mg, 115.06 μmol), N-hydroxysuccinimide (14.5 mg, 126 μmol) were dissolved in dichloromethane (2 mL) with a small amount of tetrahydrofuran (0.2 mL) as cosolvent, N,N-dicyclohexylcarbodiimide (26.1 mg, 126 μmol) was added slowly into the reaction mixture, the reaction mixture was stirred at room temperature for 1 hour. The reaction was complete by LCMS detection, the reaction mixture was filtered and the filtrate was concentrated to give the crude title compound 3-1 (110 mg), which was used directly in the next step.

[0699] Step two: Synthesis of 1-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)cyclopropane-1-carboxylic acid

[0700] Compound 3-1 (110 mg, 113.85 pmol), 1-aminocyclopropane-1-carboxylic acid (35 mg, 345 pmol), 4-dimethylaminopyridine (14 mg, 115 pmol) were dissolved in dry N,N-dimethylformamide (2 mL), N,N-diisopropylethylamine (44.59 mg, 345 pmol) was added slowly to the reaction, the reaction was stirred at room temperature for 2 hours. Saturated brine was added to the reaction, extracted with ethyl acetate and dried over anhydrous sodium sulfate to obtain the crude product. The crude product was purified by prep-HPLC [YMC-TAR column 5 pm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 7 mmol / L NH4HCO3) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 70%-80%, elution time 9 minutes] to obtain the title compound 030 (10 mg).

[0701] MS (ESI): m / z = 952.3 [M+H] + .

[0702] 1 H NMR (400 MHz, DMSO-d6) d 9.82-8.76 (m, 2H), 8.18 (s, 1H), 6.68-6.60 (m, 2H), 5.17-5.00 (m, 6H), 4.64-4.42 (m, 1H), 4.43 (d, J = 17.1 Hz, 1H), 4.22-4.09 (m, 3H), 3.79-3.68 (m, 2H), 3.51-3.40 (m, 2H), 3.21-3.13 (m, 2H), 2.89-2.76 (m, 2H), 2.46-2.40 (m, 2H), 2.16-2.05 (m, 4H), 2.05-1.96 (m, 4H), 1.96-1.88 (m, 4H), 1.67-1.50 (m, 24H), 1.19-1.11 (m, 6H), 1.07-1.00 (m, 2H), 0.88-0.80 (m, 2H).

[0703] Example 53: Synthesis of 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido)propanedioic acid (Compound 039)

[0704] Method one:

[0705] Step 1: Synthesis of 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido) propanedioic acid dimethyl ester

[0706] Compound 1-5 (100.00 mg, 115.06 μmol), 2-amino propanedioic acid dimethyl ester hydrochloride (31.69 mg, 172.52 μmol), 2-(7-azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65.11 mg, 171.23 μmol) N,N-diisopropyl ethylamine (44.61 mg, 345.17 μmol) were dissolved in N,N-dimethylformamide (1.0 mL), the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was quenched by adding water at 0 °C, then extracted with ethyl acetate (10*3 mL), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound 53-1 (90 mg).

[0707] Step 2: Synthesis of 2-((S)-2,5-bis((2R,3S)-2-((E)-4,8-dimethylnona-3,7-dien-1-yl)- 3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3-e]isoindol-8(2H)-yl)pentanamido) propanedioic acid

[0708] Lithium hydroxide (21.59 mg, 901.62 μmol) and compound 53-1 (90 mg, 90.16 μmol) were dissolved in a tetrahydrofuran (2.0 mL) and water (1.0 mL) solution, the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure, the pH value was adjusted to 6 with 2M HCl solution, concentrated under reduced pressure, the residue was purified by prep-HPLC [YMC-TAR column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 7 mmol / L NH4HCO3) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 30%-70%, elution time 10 minutes] to give the target compound 039 (13.13 mg).

[0709] MS (ESI): m / z = 970.2 [M+H]+.

[0710] 1H NMR (400 MHz, DMSO-d6) δ 9.99 - 9.52 (m, 2H), 8.29 (s, 1H), 6.65 (s, 1H), 6.61 (s, 1H), 5.11 - 5.01 (m, 6H), 4.89 - 4.83 (m, 1H), 4.51 (d, J = 17.1 Hz, 1H), 4.33 (d, J = 8.2 Hz, 1H), 4.18 - 4.09 (m, 3H), 3.77 - 3.71 (m, 2H), 3.51 - 3.43 (m, 2H), 2.87 - 2.77 (m, 2H), 2.48 - 2.39 (m, 2H), 2.20 - 1.68 (m, 14H), 1.67 - 1.44 (m, 24H), 1.19 - 1.09 (m, 6H).

[0711] Method two:

[0712] Using compound 1-1 as raw material, reference was made to example 1 to replace compound 1-1 to prepare compound 039. MS (ESI): m / z = 970.2 [M+H] + .

[0713] Reference was made to the synthesis method of the corresponding example in the table, and the corresponding starting material was replaced to synthesize the following compounds:

[0714] Biological activity and related property test examples

[0715] The compounds in the following test examples were prepared according to the method of the above examples of the present disclosure.

[0716] Test example 1 compound on the activation effect of plasminogen

[0717] ​Human Glu-Plasminogen (Enzyme research, Catalog No.: HPG2001) was prepared in buffer (50 mM Tris-HCl + 100 mM NaCl + 0.01% Tween-80, pH 7.4) at a stock concentration of 150 nM (3×), urokinase-type plasminogen activator (uPA) (Sigma-Aldrich, Catalog No.: U4010) was prepared at a stock concentration of 15 IU / ml (3×), and the plasmin substrate D-Val-Leu-Lys-pNA (Sigma-Aldrich, Catalog No.: V0882) was prepared at a stock concentration of 300 μM (3×). The working solution of the test compound was 750 μM (3×) (the compound storage solvent was DMSO). 20 μl 3× plasminogen (Human Glu-Plasminogen), 20 μl test compound (DMSO concentration in 3× working solution is 7.5%), and a control group (no compound added, replaced with 7.5% DMSO / buffer) were added, mixed and incubated at room temperature for 10 minutes. Then 20 μl (urokinase-type plasminogen activator (uPA) + D-Val-Leu-Lys-pNA) mixed working solution (3×) was added, mixed and added to a 96-well plate (Corning, catalog number: 3599), and kinetic detection was performed using an MD microplate reader (model: MD spectra Max i3x) with a detection wavelength of 405 nm and detection once every 1 minute. According to the absorbance value (OD) at a wavelength of 405 nm, the kinetics of the reaction mixture was analyzed. 405nm ) and the square of time (t 2 ) to draw a curve and use the slope of the curve to calculate the initial reaction rate, that is:

[0718] V initial =10 3 ×(OD t2 -OD t1 ) / (t2-t1) 2 , t2 and t1 are two time points with an interval of 2 minutes (OD t2 That is, OD at 4 minutes 405nm Value, OD t1 That is, OD at 2 minutes 405nm value).

[0719] The activation fold of the test compound = the initial rate of the test compound / the initial rate of the control group, and the following results were obtained (Table 1).

[0720] Table 1. Potency of compounds in activating plasminogen

[0721] Test Example 2 In vitro thrombolytic activity of the compound on rat thrombus rings

[0722] Freshly collect 500 μL SD rat (Vantianlihua) whole blood in sodium citrate anticoagulant tube (Kangshi, item number: B0980889) and stand at 4°C for standby. Prepare rat thrombus coagulation accelerator, which is a mixture of CaCl2 (Shanghai Shenguo, item number: A600506) and Thrombin (Sigma, item number: T4648-1KU), so that the final concentration of CaCl2 is 10 mM and the final concentration of Thrombin is 1 U / ml; dilute the collected rat whole blood with PBS solution (Hyclone, item number: SH30256.01) by 2 times to obtain rat whole blood diluent. In a 96 flat-bottom plate (Corning, item number: 3599), prepare rat thrombus ring, set groups as follows: 1) solvent control group (thrombus ring + PBS), 2) drug administration group (thrombus ring + compound), 3) maximum lysis group (anticoagulated whole blood without coagulation accelerator), three replicates per group. The rat thrombus ring preparation process is as follows: use a pipette gun to draw a circle along the edge of each hole in the 96-well plate, and then add thrombus coagulation accelerator (5 μL / hole), rat whole blood diluent (15 μL / hole) in turn, try to cover evenly to form a complete closed loop; place the thrombus ring in a 37°C incubator and stand for 1 hour to form a coagulated thrombus ring. During thrombus coagulation, prepare the working solution of the test compound with PBS solution (Hyclone, item number: SH30256.01) so that the final concentration of the compound is 50 μM and 100 μM. After the thrombus ring is formed, add the test compound (130 μL / hole) to the drug administration group, add PBS solution (130 μL / hole) to the blank group, and add rat whole blood diluent (15 μL / hole) + PBS solution (135 μL / hole) to the maximum lysis group. Place the 96-well plate in an enzyme marker (model: MD spectra Max i3x) for kinetic detection, detection wavelength 570 nm, 2 minutes detection once, continuous monitoring for 2 hours. Use Graphpad prism software to draw a curve according to the change of absorbance value (ΔOD 570 nm 570 nm) at 2 hours under the wavelength of 570 nm and time (min). 570nm The ΔOD 570nm indicates the in vitro thrombolytic activity of the test compound, the larger the ΔOD # , the stronger the thrombolytic activity of the test compound. Use Two-way ANOVA analysis for statistical difference, compared with the PBS treatment group, *p<0.05, ***p<0.01, ****p<0.001; compared with the control molecule treatment group, ##p<0.01. The results are shown in Figure 1. Compared with the PBS treatment group, compound 001 has significant thrombolytic activity in vitro in a dose-dependent manner; and the thrombolytic activity of compound 001 at concentrations of 50 μM and 100 μM is superior to that of the control molecule, with significant differences.

[0723] The structure of the control molecule is shown below (the control molecule mentioned in all test examples in the present disclosure is SMTP-7L prepared by microbial fermentation according to Example 2 of patent JP2004-224738A):

[0724] Test Example 3 Activation of plasminogen by the compound

[0725] The working solution of Human Glu-Plasminogen (Enzyme research, item number: HPG2001) was prepared using a buffer (50 mM Tris-HCl + 100 mM NaCl + 0.01% Tween-80, pH 7.4) at a concentration of 250 nM (5x), the working solution of urokinase-type plasminogen activator (uPA) (Sigma-Aldrich, item number: U4010) was prepared at a concentration of 125 IU / mL (2.5x), the working solution of plasmin substrate H-Val-Leu-Lys-pNA·2HCl (Absin, item number: abs45133644) was prepared at a concentration of 250 μM (2.5x), and the working solution of the test compound was prepared at a concentration of 625 μM (2.5x) or 500 μM (2.5x) or 400 μM (2.5x) (the solvent of the compound stock solution was DMSO). 40 μL / well (urokinase-type plasminogen activator (uPA) + H-Val-Leu-Lys-pNA·2HCl) mixed working solution (2.5x) was added to a 96-well enzyme plate (Sybio, item number: 100096H), 20 μL / well 5x plasminogen (Human Glu-Plasminogen) was added, followed by 40 μL / well test compound (2.5x working solution, DMSO concentration was 6.25%), and a control group (without compound, replaced with 6.25% DMSO / buffer) was set. After mixing, the kinetics was detected using an MD microplate reader (model: MD spectra Max i3x) at 37°C, the detection wavelength was 405 nm, and the detection was performed once every minute for 30 min. The curve was plotted according to the absorbance value (OD 405nm ) at 405 nm wavelength versus time 2 ), and the initial rate of the reaction was calculated using the slope of the curve, i.e. V initial = 10 3 × (OD t2 - OD t1(t2-t1) 2 t2 and t1 are two time points with an interval of 2 min (for example: 2-4 min initial rate calculation, i.e. OD t2 i.e. OD at 4 min 405nm value, OD t1 i.e. OD at 2 min 405nm value). Activation fold of the test compound = initial rate of the test compound / initial rate of the control group, to obtain the following results (Table 2).

[0726] Table 2. Effect of the compound on activation of plasminogen

[0727] Test Example 1 and Test Example 3 show that the compound of the present embodiment has significant pro-plasminogen activation activity compared with the control group.

[0728] Test Example 4: Detection of pharmacokinetic properties of the compound of the present disclosure

[0729] Test principle: Using LC / MS / MS method, the drug concentration in the plasma of rats at different time points after intravenous administration (10% bolus + 90% infusion) of the compound of the present disclosure was determined. The pharmacokinetic behavior of the compound of the present disclosure in rats was studied, and its pharmacokinetic characteristics were evaluated.

[0730] Test method:

[0731] 1.1 Test animals

[0732] Three healthy adult SD rats (6-8 weeks, 200-300 g), male, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (animal production license number: SCXK (Zhejiang) 2019-0001).

[0733] 1.2 Drug preparation

[0734] A certain amount of test compound was weighed and dissolved in 0.9% sodium chloride + 20mM meglumine + 1% castor oil (pH 9.0) to prepare 1 mg / mL for intravenous administration (10% bolus + 90% infusion).

[0735] 1.3 Drug administration

[0736] The rat tail vein was implanted with a retention needle, and 10% of the drug solution was first taken from the drug solution through a double-channel tube, and directly injected on one side (rapid injection within 5 seconds). After the end of the bolus injection, the other side was immediately continuously infused with the remaining drug solution for 30 min (the two stages of drug administration should be continuous and cannot be interrupted). No fasting was required before drug administration. The dose of drug administration was 10 mg / kg, and the volume of drug administration was 10 mL / kg.

[0737] 1.4 Operation

[0738] After intravenous administration (10% bolus + 90% infusion) in rats, blood samples (200 μL) were collected from the jugular vein at 5 min, 15 min, 30 min, 1 h, 1.5 h, 2 h, 3 h, 4 h, 6 h, 8 h, 24 h after administration, and the plasma was separated by centrifugation at 12000 rpm at 4°C for 5 min and stored at -20°C.

[0739] The concentration of the test compound in the plasma of rats after intravenous administration (10% bolus + 90% infusion) of different concentrations of the compound was determined: the sample was thawed at room temperature and vortexed for 1 min; 30 μL was quantitatively transferred to a 2 mL 96-well plate, 150 μL acetonitrile (containing an internal standard dexamethasone 100 ng / ml) was added for precipitation, and oscillation (4000 rpm*3 min) was performed; centrifugation (4000 rpm*15 min) was performed, and 100 μL of supernatant was transferred to a 2 mL 96-well plate, 50 μL water was added, and oscillation shaking (4000 rpm*3 min) was performed, and 10 μL was injected for analysis. LC / MS / MS conditions: mobile phase A: 5 mM ammonium acetate (water: acetonitrile = 80:20), mobile phase B: 5 mM ammonium acetate (water: acetonitrile = 20:80), chromatographic column: waters HPLC HSS T3 1.8 μm (2.1 mm*5 mm), column temperature: 40°C, flow rate: 0.6 mL / min.

[0740] 1.5 Data processing

[0741] The main pharmacokinetic parameters were calculated by non-compartment statistical moment method of Phoenix WinNonlin software.

[0742] Test results:

[0743] Table 3 Pharmacokinetic parameters of rats after single intravenous administration (10% bolus + 90% infusion) of the compound

[0744] The compound of the embodiments of the present disclosure has high drug exposure (AUC) and good PK properties.

[0745] Test Example 5: Detection of in vitro antioxidant activity of the compound by ORAC method

[0746] The ORAC (Oxygen Radical Absorbance Capacity) assay kit (Abeam, Cat: ab233473) was used to evaluate the in vitro antioxidant activity of the compounds. The kit includes a free radical generator (azo compound AAPH (2.2-azobis(2-methylpropyl imido) dihydrochloride), which needs to be prepared into an 80 mg / ml solution with PBS), a dilution buffer (4x, which needs to be diluted to 1x buffer with double distilled water), a fluorescent probe (100x, which is diluted 100 times to 1x fluorescent probe working solution with 1x buffer), Trolox standard, and a black 96-well flat-bottom plate. The free radical generator can destroy the fluorescent probe, causing the fluorescence intensity to decay; under the action of antioxidants, it can inhibit the fluorescence decay mediated by free radicals, so the quantitative analysis of the area under the fluorescence decay curve can be used as an indicator to measure the activity of antioxidants. The experimental group settings are as follows: 1) test compound group (compound + free radical generator + fluorescent probe), 2) probe control group (only add fluorescent probe), 3) untreated group (solvent control (no compound, replaced with 3% DMSO / 1x buffer) + free radical generator + fluorescent probe). After the reagents are equilibrated to room temperature, prepare the free radical generator working solution (80 mg / ml AAPH solution), the fluorescent probe working solution (1x), and the test compound working solution (8x, final concentration 150 μM or 250 μM) according to the instructions. The test compound storage solution is DMSO, and the test compound working solution is prepared with 1x buffer, with a DMSO concentration of 24% in the 8x working solution. In the black 96-well flat-bottom plate, add the test compound working solution (25 μL / well) and the fluorescent probe working solution (1x) (150 μL / well) in sequence, mix well, and incubate in a 37°C incubator for 30 min. After 30 min of incubation, add the free radical generator working solution (25 μL / well) to the plate and mix well. Place the 96-well plate in a microplate reader (model: MD spectra Max i3x) for kinetic detection, with an excitation / emission wavelength of 480 / 520 nm, detecting every 2 min, and continuously monitoring for 1 hour. According to the fluorescence signal value RFU, calculate the area under the fluorescence curve by the following formula:

[0747] AUC = (RFU0 + RFU2 + RFU4… + RFU 60 ) / RFU0

[0748] RFU0 represents the relative fluorescence intensity value at the starting point of detection, and RFU x represents the relative fluorescence intensity value at the corresponding time (RFU4 is the relative fluorescence intensity at 4 min). The antioxidant activity of the test compound = 100% x (AUC 待测化合物 -AUC 未处理组 ) / (AUC 探针对照组 -AUC未处理组 ). The antioxidant activity of the test compounds was plotted as a bar graph using GraphPad Prism software, and the statistical differences were analyzed using Ordinary one-way ANOVA compared with the control molecule, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The results are shown in Figure 2, Figures 2A and 2B are bar graphs of the antioxidant activity of the compounds in each group at a concentration of 150 μM. As can be seen from Figures 2A and 2B, at a concentration of 150 μM, the antioxidant activity of compounds 030, 039, 006, 062 and 057 is better than that of the control molecule and has a statistical difference. Figure 2C is a bar graph of the antioxidant activity of the compounds in each group at a concentration of 250 μM. As can be seen from Figure 2C, at a concentration of 250 μM, the antioxidant activity of compound 049 is better than that of the control molecule and has a statistical difference.

[0749] The compounds of the present disclosure have good antioxidant activity, which can avoid the risk of nerve damage caused by the penetration of intravascular obstruction.

[0750] Test Example 6 In vivo pharmacodynamic test of the control molecule in a rat autologous blood brain thrombosis stroke model

[0751] 1. Preparation of a cerebral infarction model

[0752] The autologous blood brain thrombosis stroke model was prepared according to the literature method (Brain Research 766 1997 83-92). The drug was administered 4 h after the autologous thrombus blocked the middle cerebral artery, simulating the use of drugs in patients with high risk of hemorrhage during the thrombolytic window period.

[0753] 1.1 Preparation of autologous blood thrombus

[0754] A 40 cm long blood sample was extracted from the femoral artery of a rat using a PE catheter, and then the catheter containing the blood was placed in a room temperature environment for 2 h, and then placed in a 4°C environment for 22 h. The PE tube containing the thrombus was cut into several segments of 3 cm long, and then the thrombus was blown from the segments into a dish containing normal saline using a syringe containing normal saline, and the thrombus and serum were separated. Then the thrombus was sucked into another PE tube and washed into another dish filled with normal saline. This step was repeated several times until a single solid 3 cm long thrombus was obtained. Finally, the separated thrombus was sucked into a PE tube for use.

[0755] 1.2 Preparation of a rat autologous blood brain thrombosis stroke model

[0756] Male SD rats were anesthetized with isoflurane gas and fixed on the operating table in a supine position. The skin was incised along the median line of the neck, and the common carotid artery, external carotid artery, and internal carotid artery on one side were separated. A small incision was made at the external carotid artery, and a PE catheter with a 100 μl syringe (filled with normal saline) at the end was inserted into the external carotid artery, inserted into the bifurcation of the common carotid artery in a retrograde manner, and gently inserted into the internal carotid artery about 2.5 cm deep, about 2-3 mm from the start of the internal carotid artery. Then, about 5 μl of the syringe was gently pushed, and the thrombus was slowly injected into the internal carotid artery for about 10 s. Five minutes after the injection was completed, the catheter was withdrawn from the blood vessel, and the external carotid artery was ligated. The wound was sutured, and the modeling was completed.

[0757] 2. Grouping and administration

[0758] Neurological function scoring was performed 4 h after the end of modeling, and rats with a score of ≥2 were considered to have successful modeling. The successfully modeled rats were divided into a model control (Model) group, a control molecule bolus+infusion group (dose of 10 mg / kg), and a control molecule bolus group (dose of 10 mg / kg), with 8 rats in each group. The control molecule bolus+infusion group and the control molecule bolus group were given the control molecule (10 mg / ml, 0.9% NaCl+20 mM meglumine+1% polyoxyethylene hydrogenated castor oil 60, adjusted to pH 9.0 with hydrochloric acid). Intravenous administration was performed immediately after 4 h of modeling. The control molecule bolus+infusion group was given a 10% bolus injection of the drug solution within 5-10 s, and the remaining 90% was infused within 30 min. The control molecule bolus group was given a bolus injection of the entire drug solution within 5-10 s. The end of the test was 24 h after administration.

[0759] 3. Neurological function scoring

[0760] Animal behavior disorders were observed and scored before administration. The scoring criteria were as follows:

[0761] 4. Brain hemorrhage scoring and cerebral infarction range determination

[0762] After 24 h of administration, the rats were anesthetized, the left ventricle was perfused with normal saline, and then the brain was removed on an ice platform. After being stored at -20°C for 5 min, the brain was cut into 6 pieces of the same thickness (2 mm), the hemorrhage in the brain slices was observed, and the severity of the hemorrhage was scored according to the following criteria:

[0763] 2% TTC (chloride triphenyl tetrazolium) solution was incubated at 37°C in the dark for 30 min. After staining was completed, the reaction was terminated by fixing in 4% formaldehyde solution. The ischemic area was white, and the normal area was red. The infarction volume of the brain slices was calculated using Image J, and the average value was taken.

[0764] Percentage of brain infarction volume = (volume of contralateral hemisphere - volume of normal hemisphere of infarction side) / volume of contralateral hemisphere x 100%.

[0765] 5. Experimental results

[0766] The experimental results are shown in Table 4, Figure 3 and Figure 4.

[0767] Table 4: Brain infarction volume and brain hemorrhage score * The dead animal died due to surgical operation.

[0768] Conclusion: In terms of brain infarction volume, from Table 4 and Figure 3, it can be seen that the control molecule can improve the brain infarction volume compared with the model group; in terms of brain hemorrhage, from Table 4 and Figure 4, it can be seen that the hemorrhage is lighter and the hemorrhage rate is lower after administration in the control molecule bolus + infusion group compared with the control molecule bolus group, and the risk of brain hemorrhage is significantly reduced.

[0769] Test Example 7: In vivo pharmacodynamic test of the compound of the present disclosure in a rat autologous blood cerebral thrombosis stroke model

[0770] 1. Preparation of cerebral infarction model

[0771] The autologous blood cerebral thrombosis stroke model was prepared according to the literature method (Brain Research 766 1997 83-92). The drug was administered 4h after the autologous thrombus blocked the middle cerebral artery, simulating the drug administration of patients with high hemorrhage risk in the thrombolysis window period.

[0772] 1.1 Preparation of autologous blood thrombus

[0773] A 40cm long blood was extracted from the femoral artery of a rat with a PE catheter, and then the catheter containing the blood was placed in a room temperature environment for 2h, and then placed in a 4°C environment for 22h. The PE tube containing the thrombus was cut into several segments of 3cm long, and then the thrombus was blown from the segments into a dish containing normal saline with a syringe containing normal saline, and the thrombus and serum were separated. Then the thrombus was sucked into another PE tube and flushed into another dish filled with normal saline. This step was repeated several times until a single solid 3cm long thrombus was obtained. Finally, the separated thrombus was sucked into a PE tube for standby.

[0774] 1.2 Preparation of rat autologous blood cerebral thrombosis stroke model

[0775] Male SD rats were fixed on the operating table in supine position after isoflurane gas anesthesia, the skin was incised along the median line of the neck, and the common carotid artery, external carotid artery and internal carotid artery on one side were separated. A small incision was made at the external carotid artery, and a PE catheter with a 100 μl syringe (filled with normal saline) at the end was inserted into the external carotid artery, inserted into the bifurcation of the common carotid artery in reverse direction, and gently inserted into the internal carotid artery about 2.5 cm deep, about 2-3 mm from the beginning of the internal carotid artery. Then gently push the syringe about 5 μl, about 10 s or so slowly inject the thrombus into the internal carotid artery. 5 min after injection, the catheter was withdrawn from the blood vessel, and the external carotid artery was ligated. Suture the wound, complete the modeling.

[0776] 2. Grouping and administration

[0777] The neurological function score was performed 4 h after the end of modeling, and ≥2 points were considered as successful modeling, and the successfully modeled rats were divided into a model control group (Model), a control molecule bolus+infusion group (dose of administration 10 mg / kg), and a compound 023 bolus group (dose of administration 10 mg / kg), 8 rats in each group. The control molecule bolus+infusion group was given the control molecule (10 mg / ml, 0.9% NaCl+20 mM meglumine+1% polyoxyethylene hydrogenated castor oil 60, hydrochloric acid to adjust pH to 9.0), and the compound 023 bolus group was given compound 023 (10 mg / ml, 0.9% NaCl+20 mM meglumine+1% polyoxyethylene hydrogenated castor oil 60, hydrochloric acid to adjust pH to 9.0). Intravenous administration was performed immediately after 4 h of modeling, and the control molecule bolus+infusion group was given a rapid bolus injection of 10% of the drug solution within 5-10 s, and the remaining 90% was infused within 30 min. The compound 023 bolus group was given a rapid bolus injection of all the drug solution within 5-10 s. The end of the test was 24 h after administration.

[0778] 3. Neurological function score

[0779] The animal behavior disorder was observed and scored before administration. The scoring criteria are as follows:

[0780] 4. Brain hemorrhage score and cerebral infarction range determination

[0781] After 24 h of administration, the rats were anesthetized, the left ventricle was perfused with normal saline, and then the brain was taken on the ice table, and after being stored at-20℃ for 5 min, the brain was cut into 6 pieces with the same thickness (2 mm), the hemorrhage in the brain slices was observed, and the score was given according to the severity of the hemorrhage, and the scoring criteria are as follows:

[0782] The ischemic area was white and the normal area was red. The infarct volume of the brain slices was calculated using Image J, and the average value was taken.

[0783] The percentage of cerebral infarction volume % = (contralateral half brain volume-normal volume on the infarction side) / contralateral half brain volume x 100%.

[0784] 5. Experimental results

[0785] The experimental results are shown in Table 5, Figure 5 and Figure 6.

[0786] Table 5: Cerebral infarction volume and cerebral hemorrhage score

[0787] Conclusion: In terms of cerebral infarction volume, from Table 5 and Figure 5, it can be seen that, compared with the model group, both compound 023 and the control molecule can improve the cerebral infarction volume; in terms of cerebral hemorrhage, from Table 5 and Figure 6, it can be seen that, at a dose of 10 mg / kg, the degree of hemorrhage and the rate of hemorrhage after administration of the compound 023 bolus group are comparable to those of the control molecule bolus+infusion group. Combined with the conclusion obtained in Test Example 6 that "compared with the control molecule bolus group, the degree of hemorrhage after administration of the control molecule bolus+infusion group is lighter, and the rate of hemorrhage is lower", it can be concluded that, compared with the control molecule bolus group, the degree of hemorrhage after administration of the compound 023 bolus group is lighter, and the rate of hemorrhage is lower, and the risk of cerebral hemorrhage is significantly reduced.

[0788] The above results suggest that compound 023 has an improving effect on the cerebral infarction volume after cerebral infarction, and the risk of hemorrhage is controllable, which may enable only intravenous bolus administration in clinical practice, greatly improving the convenience of administration and reducing the risk of stroke progression during inter-hospital transport.

[0789] Test Example 8: In vivo efficacy test of the compound of the present disclosure in a rat tMCAO model

[0790] 1. Animal information

[0791] SD rats, male, body weight: 250-280 g

[0792] 2. Drug information and grouping administration scheme

[0793] Note:

[0794] 1. Group 1, vehicle control group, given vehicle 0.9% NaCl+20mM meglumine+1% HCO solution;

[0795] 2. Group 2 and group 3 solvents are 0.9% NaCl + 20mM meglumine + 1% HCO solution, the concentration of the prepared drug is 1 mg / mL;

[0796] 3. Group 2 and group 3 are injected with all the liquid medicine through the tail vein within 5-10 seconds.

[0797] 3. Drugs and reagents

[0798] Compound, isoflurane, normal saline, 75% alcohol (for disinfection), iodophor

[0799] 4. Experimental instruments and consumables

[0800] Gas anesthesia machine, surgical fixation plate, cold light source, shaver, heat preservation pad, syringe, surgical instruments (need to be sterilized in advance) including: surgical scissors, fine scissors, tweezers 2, hemostatic forceps 1-2, needle holder, artery clamp, mouth opener, suture needle, 4-0 / 6-0 suture, absorbent cotton, MCAO wire plug (wire diameter: 0.40-0.45mm for rats)

[0801] 5.1 tMCAO model construction

[0802] (1) Before tMCAO operation, the animals were fasted for 12 hours without water, then anesthetized with isoflurane inhalation (4% induction, 2% maintenance), fixed supine, prepared skin routinely, disinfected, and kept warm during the whole operation.

[0803] (2) Make a midline incision from the neck using conventional ophthalmic scissors, and use tweezers to bluntly separate the glandular tissue and fascia of the neck, exposing the right common carotid artery (CCA), external carotid artery (ECA), and internal carotid artery (ICA).

[0804] (3) Use silk thread to slightly lift the CCA (to block blood flow and facilitate separation of the external carotid artery). Separate the main trunk of the ECA and ligate the distal and proximal ends of the ECA (leave a long thread end), with the proximal end thread end 5mm apart from the CCA bifurcation point. Cut the artery from the middle of the two ligation points, then gently pull the proximal end thread to make the cut artery consistent with the shape of the common carotid artery.

[0805] (4) Prepare the thread on the ECA residual segment, temporarily clamp the ICA with a micro-artery clamp, then cut a small opening on the residual segment of the external carotid artery, gently push the prepared plug wire into the ICA, and open the artery clamp on the ICA to make the wire plug enter the middle cerebral artery (MCA) starting end along the ICA, and fix the plug wire with the standby silk thread. From the CCA bifurcation to the resistance point (the starting point of the middle cerebral artery), the depth of the wire in the rat is about 18-20mm, and finally remove the silk thread on the CCA to restore blood flow. Suture the skin and disinfect the neck wound with iodophor.

[0806] (5) After the animal model awakens, obvious hemiplegia symptoms appear, the body tilts and turns around, which is considered as a successful modeling. 2h after ischemia, reperfusion is performed, the animal is anesthetized with isoflurane (4% induction, 2% maintenance), the neck suture incision is opened, and the ECA is gradually pulled out until the thread plug completely leaves the ECA, and the ECA stump is ligated with an electrocoagulator or suture.

[0807] (6) The neck incision is sutured again, iodophor is applied for disinfection, and single-cage feeding is performed.

[0808] 5.2 Dosing regimen

[0809] During the operation, animals with abnormal conditions due to anesthesia, operation, etc. must be removed. Immediately after cerebral ischemia-reperfusion, the drug is administered once.

[0810] 6. Detection index

[0811] 1) Percentage of cerebral infarction volume

[0812] 24h after ischemia, euthanasia is performed on the surviving rats in each group, the heart is perfused with normal saline, and the brain tissue is taken. 2% red tetrazolium (TTC) is used for staining, and the photograph is analyzed using Image J image analysis software to calculate the percentage of cerebral infarction volume.

[0813] Among them: the percentage of whole brain cerebral infarction volume % = ((contralateral half brain volume-normal volume on the infarction side) / contralateral half brain volume x 100%) / 2 Note: The percentage of cerebral infarction volume mentioned in this test example is the percentage of whole brain cerebral infarction volume

[0814] 2) Severity score of cerebral hemorrhage

[0815] Normal saline is used for body circulation perfusion to take brain sections, the hemorrhage in the brain sections is observed, and the score is given according to the severity of the hemorrhage, and the scoring criteria are as follows:

[0816] No hemorrhage 0 points, no trace of hemorrhage

[0817] Mild 1 point, no obvious bleeding point, with slight bleeding trace, blood mark is light pink

[0818] Mild 2 points, no obvious bleeding point, with obvious bleeding trace, blood mark is light pink

[0819] Moderate 3 points, a small amount of bleeding point, or obvious bleeding trace, blood mark is pink

[0820] Moderate 4 points, obvious bleeding point, or obvious bleeding trace with a larger area, blood mark is pink

[0821] Severe 5 points, more bleeding points, and obvious bleeding trace with a larger area, blood mark is magenta

[0822] Severe 6 points, more bleeding points, and obvious and large area of bleeding marks, bloodstain is dark red

[0823] Pharmacodynamic endpoints

[0824] Percentage of cerebral infarction volume

[0825] Cerebral hemorrhage severity score

[0826] 7. Statistical analysis method

[0827] The percentage of cerebral infarction volume was statistically analyzed by ONE WAY ANOVA analysis method using GraphPad Prism 8.0, and the cerebral hemorrhage severity score was statistically analyzed by non-parametric test using GraphPad Prism 8.0, and p<0.05 was considered to be significantly different in all analyses.

[0828] 8. Experimental results

[0829] The experimental results are shown in Figures 7-10, in which * represents p<0.05, and ns represents no significant difference.

[0830] The experimental results show that, in terms of the percentage of cerebral infarction volume, as can be seen from Figures 7 and 8, compared with the solvent control group, the compound 057 bolus group can significantly reduce the percentage of cerebral infarction volume, and no obvious improvement effect of the control molecule bolus group on the percentage of cerebral infarction volume is found. In terms of the degree of cerebral hemorrhage, as can be seen from Figures 9 and 10, compared with the control molecule bolus group, the hemorrhage degree of the compound 057 bolus group is lighter, and the risk of cerebral hemorrhage is significantly reduced. The above results suggest that the compound 057 has an improvement effect on the cerebral infarction volume after cerebral infarction, and the risk of hemorrhage is controllable, which can realize clinical intravenous bolus administration, greatly improve the convenience of drug administration, and reduce the risk of stroke progression during inter-hospital transport.

[0831] In addition, we observed that in the tMCAO pharmacodynamic model of rats, compared with the control molecule bolus+infusion group (the administration method is: first, 10% drug solution is rapidly injected through the jugular vein, and then the remaining 90% drug is slowly infused within 30 min), the hemorrhage score of the control molecule bolus group (the administration method is: all drug solution is rapidly injected through the tail vein within 5-10 s) is significantly improved.

[0832] Test Example 9: 14-day intravenous injection toxicity test of rats

[0833] 1) Purpose of the test: This test detects and compares the potential toxicity of compound 023 and the control molecule by intravenous injection of rats for 14 consecutive days.

[0834] 2) Test Method: SD rats were randomly divided into 3 groups, and the toxicity of Compound 023 and the control molecule was investigated by intravenous injection of the drug once a day for 14 consecutive days. There were 3 animals per group for each gender. The solvent control group was only given the control preparation. Random grouping was performed in Provantis according to the body weight of the animals. The dose design is shown in the following table:

[0835] Note: Group 1 solvent is 0.9% NaCl + 20mM meglumine + 1% polyoxyethylene hydrogenated castor oil 60, pH adjusted to 9.0 with hydrochloric acid; Group 2 and Group 3 solvents are 0.9% NaCl + 20mM meglumine + 1% polyoxyethylene hydrogenated castor oil 60, pH adjusted to 9.0 with hydrochloric acid.

[0836] 3) Toxicity evaluation index: clinical pathology examination was performed on Day 7 and Day 15. The results are shown in the following table:

[0837] 4) Conclusion:

[0838] Compared with the control molecule, Compound 023 has a smaller degree of change in hematological parameters and a smaller effect on white blood cells and red blood cells, indicating that Compound 023 has lower blood toxicity and higher safety compared with the control molecule.

[0839] Test Example 10: 14-day intravenous injection toxicity test in dogs

[0840] 1) Test Purpose: Beagle dogs were given Compound 023 and the control molecule by intravenous injection, once a day for 2 consecutive weeks, without a recovery period, to compare the potential toxicity of the two.

[0841] 2) Test Method: 6 Beagle dogs (3 / sex, general level, body weight: 6-11 kg at grouping; age: about 10-12 months at the start of dosing) were randomly divided into 3 groups, 1 male and 1 female in each group. A solvent control group, a Compound 023 group and a control molecule group were set up respectively; the experimental animals were given Compound 023 preparation, solvent (control) or control molecule preparation by tail vein injection once a day; the solvent control group was only given the control preparation. 2 animals were dissected per group on the dissection day, half male and half female.

[0842] The dose design is shown in the following table:

[0843] Note: Group 1 solvent is 0.9% NaCl + 20mM meglumine, pH adjusted to 9.0 with hydrochloric acid;

[0844] Group 2 and Group 3 solvents are 0.9% NaCl + 20mM meglumine, pH adjusted to 9.0 with hydrochloric acid.

[0845] 3) Toxicity evaluation index: gross anatomy.

[0846] 4) Conclusion: The control molecule 9 mg / kg group observed changes in the color of the colon - multifocal dark red in gross anatomical gross lesions, while the compound 023 15 mg / kg group did not see changes in the color of the colon, suggesting that the risk of bleeding of compound 023 is lower than that of the control molecule.

[0847] Test Example 11 Fluorescence method for detecting in vitro anti-inflammatory activity of compounds

[0848] The experiment uses soluble epoxide hydrolase inhibitor screening assay kit (Cayman, item number: 10011671) to evaluate the in vitro anti-inflammatory activity of the compound based on fluorescence method. The kit uses (3-phenyl-oxy)-acetic acid cyanide-(6-methoxy-naphthalen-2-yl)-methyl ester (PHOME) as the substrate. When the epoxide part of PHOME is hydrolyzed by epoxide hydrolase, intramolecular cyclization occurs and cyanohydrin is released, and its decomposition product 6-methoxy-2-naphthaldehyde can be detected by fluorescence signal under excitation 330 nm / emission 465 nm. The kit includes human recombinant sEH enzyme, sEH reaction buffer (10×), sEH substrate, black 96 flat-bottom plate, 96-well plate sealing film. Dilute the sEH reaction buffer (10×) 10 times with double distilled water to obtain 1×sEH reaction buffer. Dilute the human recombinant sEH enzyme 50 times with 1×sEH reaction buffer pre-cooled at 4 degrees to obtain sEH enzyme working solution, and place it on ice for use. The experimental group is set as follows: 1) test compound group (compound working solution + sEH enzyme working solution + sEH substrate), 2) High control (solvent control, i.e. final concentration 3% DMSO / reaction buffer) + sEH enzyme working solution + sEH substrate), 3) Low control (final concentration 3% DMSO / buffer + sEH substrate). After the reagents are equilibrated to room temperature, prepare the sEH substrate PHOME working solution (3×, 0.75 μM) and the test compound working solution (3×, working solution concentration 90 μM / 9% DMSO) using 1×sEH reaction buffer according to the instructions. In the black 96 flat-bottom plate, add 60 μL / well of the test compound working solution (3×) and 60 μL / well of the sEH enzyme working solution (3×) in turn, mix well, and then place at room temperature for 5 min. After the end, add 60 μL / well of the sEH substrate working solution (3×) to the plate under light-proof conditions, and mix well. Place the 96-well plate in the enzyme marker (model: MD spectra Max i3x) for kinetic detection, with excitation / emission wavelength of 330 / 465 nm, detection every 2 min, continuous monitoring for 40 min, and reading of the fluorescence signal value RFU. Take time as X axis and RLU value as Y axis, and calculate the area under the curve AUC by Graphpad Prism software. The anti-inflammatory activity of the test compound is calculated as follows: inhibition rate % = 100% × (AUC high control-low control-AUC 待测化合物组-low control ) / AUC high control-low control. The anti-inflammatory activity of the compound is shown in the following table.

[0849] C-sEH anti-inflammatory activity of the compound (10 μM)

[0850] The compound of the embodiments of the present disclosure has good C-sEH anti-inflammatory activity.

Claims

1. A compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein is a single bond or a double bond; a, b, c and d are each independently selected from a single bond or a double bond; Q 1 and Q 2 are each independently selected from O or S; R 1 selected from COOH, -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9 , amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, tetrazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, C(O)N(C1-C6alkyl)2, or -5-10 membered heteroarylene-COOH, which amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, tetrazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, C(O)N(C1-C6alkyl)2, or -5-10 membered heteroarylene-COOH is optionally substituted with R 1a ; R 1a selected from halogen, COOH, S(O)2OH, P(O)(OH)2, cyano, hydroxy, imidazolyl, tetrazolyl, OC(O)NHCi-C6alkyl, Ci-C6alkoxy, amino, NH(Ci-C6alkyl) or N(Ci-C6alkyl)2, said hydroxy, imidazolyl, tetrazolyl, OC(O)NHCi-C6alkyl, Ci-C6alkoxy, amino, NH(Ci-C6alkyl) or N(Ci-C6alkyl)2being optionally substituted with R 1b substituted; R 1b selected from COOH, S(O)2OH or P(O)(OH)2; M is selected from O, NH or NR M ; T is selected from (CH2) n , (CH2) n C(O)NH(CH2) m , phenylene, 4-12 membered heterocyclylene or C3-C6cycloalkylene, said (CH2) n , (CH2) n C(O)NH(CH2) m , phenylene, 4-12 membered heterocyclylene or C3-C6cycloalkylene is optionally substituted with R T ; R T selected from halogen, COOH, C1-C6alkyl, amino, or C3-C6cycloalkyl, said C1-C6alkyl, amino, or C3-C6cycloalkyl optionally substituted with R 10 10 cycloalkyl optionally substituted with R Ta ; or, two R T and their respective attached atoms together form a C3-C6cycloalkane ring or a 4-12 membered heterocyclic ring, said C3-C6cycloalkane ring or 4-12 membered heterocyclic ring optionally substituted with R 14 ;​ R 14 selected from halogen, COOH, C1-C6 alkyl or amino; R Ta selected from halogen, -S-Ci-C6alkyl, Ci-C6alkoxy, SH, hydroxyl, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, COOH, imidazolyl, imidazolyl substituted with -SH, phenyl, indolyl, tetrazolyl, NHC(=NH)NH2, or C(O)NH2; R M is selected from C1-C6 alkyl, wherein the C1-C6 alkyl is optionally replaced by R Ma Replacement; or R M With R T and the atoms to which they are attached together form a 4-12 membered heterocyclic ring, wherein the 4-12 membered heterocyclic ring is optionally replaced by R 13 replace; R 13 selected from hydroxyl, cyano, SH, amino, carboxyl, halogen or C1-C6 alkyl; R Ma selected from COOH, hydroxyl or n is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8; m is selected from 1, 2, 3, 4, 5, 6, 7 or 8; R 2 and R 4 each independently is selected from OH, SH, amino, C1-C6alkoxy, -OC(O)N(C1-C6alkyl)2, -OC(O)NH(C1-C6alkyl), -OC(O)NH2, or OC(O)C1-C6alkyl; said amino, C1-C6alkoxy, -OC(O)N(C1-C6alkyl)2, -OC(O)NH(C1-C6alkyl), -OC(O)NH2, or OC(O)C1-C6alkyl is optionally substituted with R 2a ; R 2a selected from halogen, amino, COOH, NHCi-C6alkyl or N(Ci-C6alkyl)2; R 3 and R 5 each independently is selected from hydrogen, halogen, OH, SH, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, OC(O)Ci-C6alkyl or OC(O)H; R 6 selected from hydrogen or Ci-C6alkyl; said Ci-C6alkyl is optionally substituted with R 6a substituted; R 6a selected from halogen; R 7 and R 8 Each is independently selected from hydrogen, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C3-C 10 Cycloalkyl, C2-C6 alkenyl or 5-10 membered heteroaryl, the C1-C6 alkyl, C3-C 10 Cycloalkyl, C2-C6 alkenyl or 5-10 membered heteroaryl is optionally replaced by R 7a replace; R 7a selected from amino, halogen, hydroxyl, NH(Ci-C6alkyl), or N(Ci-C6alkyl)2; R 9 selected from hydroxyl, COOH, P(O)(OH)2, S(O)2OH, S(O)OH, imidazolyl, or tetrazolyl; R 11 selected from hydrogen, deuterium, halogen or Ci-C6alkyl, said Ci-C6alkyl is optionally substituted with R 11a ; or R 11 , R 5 and the carbon atom to which they are attached together form C=CH2or C=NOH; R 12 selected from hydrogen, deuterium, halogen or Ci-C6alkyl, said Ci-C6alkyl is optionally substituted with R 12a ; or R 12 , R 3 and the carbon atom to which they are attached together form C=CH2or C=NOH; R 11a and R 12a each independently is selected from halogen; R 15 and R 16 are each independently selected from hydrogen or deuterium.

2. The compound of formula (I) according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, is a double bond, a, b, c, and d are each double bonds; or is a single bond, a, b, c and d are each a single bond.

3. The compound of formula (I) according to claim 1 or 2, wherein, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. Q 1 and Q 2 are each O.

4. The compound of formula (I) according to any one of claims 1 to 3, wherein R 1 selected from -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9 , amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, or C(O)N(C1-C6alkyl)2, said amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, or C(O)N(C1-C6alkyl)2optionally substituted with R 1a ; or R 1 selected from -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9 , CH=N(C1-C6)alkyl, C1-C6alkyl, C2-C6alkenyl, or C(O)N(C1-C6alkyl)2, said CH(=N)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, or C(O)N(C1-C6alkyl)2optionally substituted with R 1a ; or R 1 selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , C1-C3 alkyl, C2-C4 alkenyl, or -5-6 membered heteroarylene-COOH, said C1-C3 alkyl, C2-C4 alkenyl, or -5-6 membered heteroarylene-COOH optionally substituted with R 1a ; or R 1 selected from -CH=NOH, P(O)(OH)2, S(O)2OH, C(O)M-T-R 9 , amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, C(O)N(C3-C6alkyl)2, or -5-10 membered heteroarylene-COOH, which amino, NHC1-C6alkyl, N(C1-C6alkyl)2, CH=N(C1-C6alkyl), imidazolyl, C(O)imidazolyl, C(O)tetrazolyl, C(O)C1-C6alkyl, C1-C6alkyl, C2-C6alkenyl, C(O)N(C3-C6alkyl)2, or -5-10 membered heteroarylene-COOH is optionally substituted with R 1a ; or R 1 selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , C1-C6alkyl, C2-C6alkenyl, or -5-10 membered heteroarylene-COOH, said C1-C6alkyl, C2-C6alkenyl, or -5-10 membered heteroarylene-COOH optionally substituted with R 1a ; or R 1 selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , C1-C3 alkyl, C2-C4 alkenyl, or -5-6 membered heteroarylene-COOH, said C1-C3 alkyl, C2-C4 alkenyl, or -5-6 membered heteroarylene-COOH optionally substituted with R 1a ; or R 1 selected from -CH=NOH, P(O)(OH)2, C(O)M-T-R 9 , C1-C3 alkyl or C2-C4 alkenyl, said C1-C3 alkyl or C2-C4 alkenyl being optionally substituted with R 1a .

5. The compound of formula (I) according to any one of claims 1 to 4, wherein R 1a selected from COOH, S(O)2OH, P(O)(OH)2, hydroxy, imidazolyl, tetrazolyl, C1-C6alkoxy, amino, NH(C1-C6alkyl), or N(C1-C6alkyl)2, said C1-C6alkoxy, amino, NH(C1-C6alkyl), or N(C1-C6alkyl)2optionally substituted with R 1b ; or R 1a selected from COOH, hydroxy, C1-C3alkoxy, NH(C1-C3alkyl), or OC(O)NHC1-C3alkyl, said C1-C3alkoxy, NH(C1-C3alkyl), or OC(O)NHC1-C3alkyl being optionally substituted with R 1b or R 1a selected from COOH, hydroxy, C1-C3alkoxy or NH(C1-C3alkyl), said C1-C3alkoxy or NH(C1-C3alkyl) being optionally substituted with R 1b substituted; and / or R 1b selected from COOH.

6. The compound of formula (I) according to any one of claims 1 to 5, wherein M is selected from O, NH or NR M ; R M is selected from C1-C6alkyl optionally substituted with R Ma ; or R M and R T , together with the atoms to which they are attached, form a 4-10 membered heterocyclic ring optionally substituted with R 13 ; or M is selected from O, NH or NR M ; R M is selected from C1-C3 alkyl, said C1-C3 alkyl being optionally substituted with R Ma ; or R M , together with the atom to which they are attached, form a 4-6 membered heterocyclic ring, said 4-6 membered heterocyclic ring being optionally substituted with R T ; or R 13 , together with the atom to which they are attached, form a 4-6 membered heterocyclic ring, said 4-6 membered heterocyclic ring being optionally substituted with R M is selected from O, NH or NR M ; R M is selected from methyl, said methyl being optionally substituted with R Ma ; or R M and R T , together with the atom to which they are attached, form an azetidinylene, thiazolidinylene, pyrrolidinylene, piperazinylene or morpholinylene group, said azetidinylene, thiazolidinylene, pyrrolidinylene, piperazinylene or morpholinylene group being optionally substituted with R 13 ; or M is selected from NH or NR M ; and / or R Ma selected from COOH or 7. The compound of formula (I) according to claim 6, wherein, ###0002### or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. R 13 selected from hydroxyl, halogen or C1-C3 alkyl; or R 13 is selected from F, hydroxyl or methyl.

8. The compound of formula (I) according to any one of claims 1 to 7, wherein M is selected from O or NH; or M is selected from O; or M is NH.

9. The compound of formula (I) according to any one of claims 1 to 8, wherein T is selected from (CH2) n , phenylene or 4-10 membered heterocyclylene, said (CH2) n , phenylene or 4-10 membered heterocyclylene is optionally substituted with R T ; or T is selected from (CH2) n , phenylene or C4-C5cycloalkylene, which (CH2) n , phenylene or C4-C5cycloalkylene is optionally substituted with R T ; or T is selected from (CH2) n (CH2) n optionally substituted with R T ; or T is selected from (CH2) n , (CH2) n C(O)NH(CH2) m , phenylene, 4-6 membered heterocyclene or C3-C6cycloalkylene, said (CH2) n , (CH2) n C(O)NH(CH2) m , phenylene, 4-6 membered heterocyclene or C3-C6cycloalkylene is optionally substituted with R T .

10. The compound of formula (I) according to any one of claims 1 to 9, wherein n is selected from 0, 1, 2 or 3; or n is selected from 0, 1 or 2; or n is selected from 1 or 2; and / or m is selected from 1 or 2.

11. The compound of formula (I) according to any one of claims 1 to 10, wherein R T selected from amino, COOH, or C1-C6 alkyl, said C1-C6 alkyl being optionally substituted with R Ta ; or, two R T , together with their attached atoms, form a C3-C6 cycloalkane ring, said C3-C6 cycloalkane ring being optionally substituted with R 14 ; or R M , together with R T , and their attached atoms, form a 4-10 membered heterocyclic ring, said 4-10 membered heterocyclic ring being optionally substituted with R 13 ; or R T selected from amino, COOH, C1-C3 alkyl or C3-C6 cycloalkyl, said C1-C3 alkyl or C3-C6 cycloalkyl being optionally substituted with R Ta ; or, two R T , together with their attached atoms, form a C3-C4 cycloalkane ring or a 4-6 membered heterocyclic ring, said C3-C4 cycloalkane ring or 4-6 membered heterocyclic ring being optionally substituted with R 14 ; or R M , together with R T , and their attached atoms form a 4-6 membered heterocyclic ring, said 4-6 membered heterocyclic ring being optionally substituted with R 13 ; or R T selected from amino, COOH, methyl or said methyl or optionally substituted with R Ta ; or, two R T and the atoms to which they are each attached come together to form a cyclopropyl, cyclobutyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperazinyl, or thietanyl, optionally substituted with R 14 ; or, two R R T selected from halogen, COOH, C1-C6alkyl, amino or C3-C6cycloalkyl, said C1-C6alkyl, amino or C3-C6cycloalkyl being optionally substituted with R Ta ; or, two R T , together with their attached atoms, form a C3-C6cycloalkane ring or a 4-6 membered heterocyclic ring, said C3-C6cycloalkane ring or 4-6 membered heterocyclic ring being optionally substituted with R 14 ; or R M , together with R T , form a 4-6 membered heterocyclic ring, said 4-6 membered heterocyclic ring being optionally substituted with R 13 .

12. The compound of formula (I) according to any one of claims 1 to 11, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R Ta selected from halogen, -S-Ci-C6alkyl, Ci-C6alkoxy, SH, hydroxyl, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, COOH, imidazolyl, tetrazolyl, NHC(=NH)NH2, or C(O)NH2; or R Ta selected from halogen, -S-Ci-C6alkyl, SH, hydroxyl, amino, COOH, imidazolyl, imidazolyl substituted with -SH, phenyl, indolyl, NHC(=NH)NH2, or C(O)NH2; or R Ta is selected from F, SCH3, SH, hydroxyl, amino, COOH, 2-imidazolyl, 2-SH-4- imidazolyl, phenyl, 3-indolyl, NHC(=NH)NH2, or C(O)NH2; or R Ta is selected from halogen, -S-Ci-C3alkyl, SH, hydroxyl, amino, COOH, imidazolyl, NHC(=NH)NH2or C(O)NH2.

13. The compound of formula (I) according to any one of claims 1 to 12, wherein R 14 selected from the group consisting of C1-C6alkyl; or R 14 selected from the group consisting of C1-C3alkyl; and / or R 15 and R 16 are both hydrogen, or R 15 and R 16 are both deuterium.

14. The compound of formula (I) according to any one of claims 1 to 13, wherein R 9 is selected from hydroxyl, COOH, P(O)(OH)2, tetrazolyl, or S(O)2OH; or R 9 is selected from COOH.

15. The compound of formula (I) according to any one of claims 1 to 14, wherein R 1 selected from C(O)M-T-R 9 .

16. The compound of claim 15, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, M is selected from O, NH or NR M , R M and R T together with the atoms to which they are attached form a 4-6 membered heterocyclic ring, which is optionally substituted with R 13 ; T is selected from (CH2) n , (CH2) n optionally substituted with R T ; R T is selected from COOH or C1-C3 alkyl optionally substituted with R Ta ; or two R T and their respective attached atoms together form a C3-C6 cycloalkane ring optionally substituted with R 14 ; n is selected from 0, 1 or 2; R Ta selected from halogen, -S-Ci-C3alkyl, SH, hydroxyl, amino, or COOH; R 9 selected from COOH.

17. The compound of formula (I) according to any one of claims 1-15, wherein R 1 selected from C(O)M(CH2) n R 9 wherein M is selected from O, NH, NCH3, or NCH2COOH; n is 0, 1, 2, 3, 4, 5, or 6; R 9 selected from hydroxyl, COOH, P(O)(OH)2, S(O)2OH, S(O)OH, or tetrazolyl; said (CH2) n optionally substituted with 1, 2, 3, or 4 substituents independently selected from halogen, COOH, C1-C6alkyl, amino, or C3-C 10 cycloalkyl, said C1-C6alkyl, amino, or C3-C 10 cycloalkyl is optionally substituted with 1, 2, or 3 substituents independently selected from halogen, -S-C1-C6alkyl, SH, hydroxyl, amino, COOH, imidazolyl, imidazolyl substituted with -SH, phenyl, indolyl, NHC(=NH)NH2, or C(O)NH2; or R 1 selected from C(O)M(CH2) n R 9 wherein M is selected from O, NH or NCH3; n is 0, 1, 2 or 3; R 9 selected from hydroxy, COOH, P(O)(OH)2, S(O)2OH, S(O)OH or tetrazolyl; said (CH2) n optionally substituted with 1 or 2 substituents independently selected from halogen, COOH, C1-C6alkyl, amino or C3-C6cycloalkyl, said C1-C6alkyl, amino or C3-C6cycloalkyl optionally substituted with 1, 2 or 3 substituents independently selected from halogen, -S-C1-C6alkyl, SH, hydroxy, amino, COOH, imidazolyl, imidazolyl substituted with -SH, phenyl, indolyl, NHC(=NH)NH2or C(O)NH2; or R 1 selected from C(O)M(CH2) n R 9 wherein M is NH; n is 1 or 2; R 9 selected from COOH, P(O)(OH)2, or S(O)2OH; said (CH2) n optionally substituted with 1 or 2 substituents independently selected from halo or C1-C6alkyl optionally substituted with 1, 2, or 3 substituents independently selected from halo, -S-C1-C3alkyl, SH, hydroxy, amino, COOH, 2-imidazolyl, 4-imidazolyl substituted with -SH, phenyl, 3-indolyl, NHC(=NH)NH2, or C(O)NH2; or R 1 selected from or R 1 is selected from C(O)M-T-R 9 wherein M is O or NH; T is phenylene, 4-12 membered heterocyclene or C3-C6 cycloalkylene, said phenylene, 4-12 membered heterocyclene or C3-C6 cycloalkylene being optionally substituted by 1 or 2 substituents selected from halogen or C1-C6 alkyl; R 9 is selected from COOH, P(O)(OH)2 or S(O)2OH; or R 1 is selected from C(O)M-T-R 9 wherein M is O or NH; T is phenylene, 4-6 membered heterocyclene or C3-C6cycloalkylene, which phenylene, 4-6 membered heterocyclene or C3-C6cycloalkylene is optionally substituted with 1 or 2 substituents selected from halogen or C1-C3alkyl; R 9 is COOH; or R 1 selected from or R 1 is selected from C(O)M-T-R 9 wherein M is NR M , T is (CH2) T substituted with R n , n is 1, 2 or 3, R M and R T together with the atoms to which they are attached form a 4-12 membered heterocyclic ring, which is optionally substituted with 1 or 2 substituents independently selected from halogen, OH or C1-C6 alkyl, R 9 is selected from COOH, P(O)(OH)2 or S(O)2OH; or R 1 is selected from C(O)M-T-R 9 wherein M is NR M , T is (CH2) T substituted with R n , n is 1 or 2, R M and R T together with the atoms to which they are attached form a 4-6 membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O or S, and optionally substituted with 1 or 2 substituents independently selected from halogen, OH or C1-C6 alkyl, R 9 is selected from COOH; or R 1 is selected from C(O)M-T-R 9 wherein M is NR M , T is (CH2) T substituted with R n , n is 1 or 2, R M and R T together with the atoms to which they are attached form an azetidine, piperazine, thiazolidine, pyrrolidine or morpholine, said azetidine, piperazine, thiazolidine, pyrrolidine or morpholine being optionally substituted with 1 or 2 substituents independently selected from halogen, OH or C1-C3alkyl, R 9 is selected from COOH; or R 1 selected from or R 1 is selected from C(O)M-T-R 9 wherein M is O or NH, T is C(R T )2 or CH2C(R T )2, and two R T and the C atom to which they are attached together form a C3-C6 cycloalkane ring or a 4-12 membered heterocyclic ring, which is optionally substituted with 1 or 2 substituents independently selected from halogen and C1-C6 alkyl, R 9 is selected from COOH, P(O)(OH)2, or S(O)2OH; or R 1 is selected from C(O)M-T-R 9 wherein M is O or NH, T is C(R T )2 or CH2C(R T )2, and two R T and the C atoms to which they are attached together form a cyclopropane, cyclobutane, azetidine, oxetane, tetrahydrofuran, tetrahydropyran or piperidine, which is optionally substituted with 1 or 2 substituents independently selected from F, CH3 or C2H5, R 9 is selected from COOH; or R 1 selected from or R 1 selected from C(O)M(CH2) n C(O)NH(CH2) m R 9 wherein M is selected from O, NH or NCH3, n is selected from 1, 2, 3, 4, 5 or 6, m is selected from 1, 2, 3, 4, 5 or 6, R 9 is selected from hydroxyl, COOH, P(O)(OH)2, S(O)2OH, S(O)OH or tetrazolyl; said (CH2) n C(O)NH(CH2) m optionally substituted with 1, 2, 3 or 4 substituents independently selected from halogen or C1-C6alkyl optionally substituted with 1, 2 or 3 substituents independently selected from halogen, hydroxyl, amino or imidazolyl; or R 1 selected from C(O)M(CH2) n C(O)NH(CH2) m R 9 wherein M is NH, n is selected from 1 or 2, m is 1 or 2, R 9 selected from COOH; said (CH2) n C(O)NH(CH2) m optionally substituted with 1 or 2 substituents independently selected from halo or C1-C3alkyl optionally substituted with 1, 2, or 3 substituents independently selected from halo, hydroxy, amino, or 4-imidazolyl; or R 1 selected from 18. The compound of formula (I) according to any one of claims 1-14, wherein R 1 selected from or R 1 selected from 19. The compound of formula (I) according to any one of claims 1-18, wherein R 2 and R 4 each independently is selected from SH, amino, C1-C6alkoxy, -OC(O)N(C1-C6alkyl)2, -OC(O)NH(C1-C6alkyl), -OC(O)NH2, or OC(O)C1-C6alkyl; said amino, C1-C6alkoxy, -OC(O)N(C1-C6alkyl)2, -OC(O)NH(C1-C6alkyl), -OC(O)NH2, or OC(O)C1-C6alkyl is optionally substituted with R 2a ; or R 2 and R 4 each independently is selected from SH, amino, C1-C3alkoxy, OC(O)C1-C3alkyl, said C1-C3alkoxy, OC(O)C1-C3alkyl being optionally substituted with R 2a ; or R 2 and R 4 each independently is selected from OH, SH, amino, C1-C6alkoxy or OC(O)C1-C6alkyl, said amino, C1-C6alkoxy or OC(O)C1-C6alkyl being optionally substituted with R 2a ; or R 2 and R 4 each independently is selected from OH, SH, amino, C1-C3alkoxy or OC(O)C1-C3alkyl, said amino, C1-C3alkoxy or OC(O)C1-C3alkyl being optionally substituted with 1 or 2 substituents selected from halogen, COOH or N(C1-C3alkyl)2; and / or R 2a selected from halogen, COOH or N(C1-C3alkyl)2; or R 2 and R 4 each independently is selected from OH, SH, amino, OCHF2, or R 2 and R 4 are each independently selected from OH.

20. The compound of formula (I) according to any one of claims 1-19, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 5 each independently is selected from hydrogen, halogen, SH, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, OC(O)Ci-C6alkyl, or OC(O)H; or R 3 and R 5 each independently is selected from amino, OC(O)C1-C3alkyl or OC(O)H; or R 3 and R 5 each independently is selected from OH, amino, OC(O)C1-C6alkyl, or OC(O)H; or R 3 and R 5 each independently is selected from OH, amino, OC(O)CH3, or OC(O)H; or R 3 and R 5 are each independently selected from OH.

21. The compound of formula (I) according to any one of claims 1-20, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R 6 is selected from hydrogen or C1-C6alkyl; or R 6 is selected from C1-C3alkyl; or R 6 is selected from hydrogen or methyl.

22. The compound of formula (I) according to any one of claims 1-21, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R 7 and R 8 each independently is selected from hydrogen, halogen, cyano, carboxyl, C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-6 membered heteroaryl, said C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, or 5-6 membered heteroaryl optionally substituted with R 7a ; or R 7 and R 8 each independently is selected from hydrogen, halogen, cyano, carboxyl, C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl, said C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl optionally substituted with 1, 2, or 3 substituents independently selected from halogen, OH, and N(C1-C3alkyl)2; or R 7 and R 8 each independently is selected from halogen, hydroxyl, C1-C6alkyl or C3-C6cycloalkyl, said C1-C6alkyl or C3-C6cycloalkyl optionally being substituted with R 7a ; or R 7 and R 8 each independently is selected from halogen, cyano, carboxyl, C1-C3 alkyl, C3-C6 cycloalkyl, C2-C4 alkenyl or 5-membered heteroaryl, said C1-C3 alkyl, C3-C6 cycloalkyl, C2-C4 alkenyl or 5-membered heteroaryl optionally being substituted with R 7a ; and / or R 7a selected from amino, NH(Ci-C3alkyl), or N(Ci-C3alkyl)2; or R 7a selected from R 7a selected from halogen, hydroxyl or N(C1-C3alkyl)2; or R 7 and R 8 each independently is selected from hydrogen, bromo, cyano, methyl, CF3, CHF2, CH2OH, CH2N(CH3)2, vinyl, COOH, cyclopropyl, or pyrazolyl; or R 7 and R 8 are each independently selected from hydrogen.

23. The compound of formula (I) according to any one of claims 1-22, wherein R 11 selected from deuterium, halogen, or Ci-C6alkyl optionally substituted with R 11a ; or R 11 , R 5 and the carbon atom to which they are attached together form C=CH2or C=NOH; or R 11 , R 5 and the carbon atom to which they are attached form C=NOH; or R 11 selected from hydrogen.

24. The compound of formula (I) according to any one of claims 1-23, wherein R 12 selected from deuterium, halogen, or Ci-C6alkyl optionally substituted with R 12a ; or R 12 , R 3 and the carbon atom to which they are attached together form C=CH2or C=NOH; or R 12 , R 3 and together with the carbon atom to which they are attached form C=NOH; or R 12 selected from hydrogen.

25. The compound of formula (I) according to any one of claims 1-24, wherein R 1 is COOH, and R 6 is methyl; or R 1 is COOH, and R 3 and R 5 are the same and are amino, OC(O)C1-C6alkyl, or OC(O)H; or R 1 is COOH, and a, b, c, and d are each a single bond; or R 1 is COOH, and R 7 and R 8 are the same and are hydrogen, halogen, cyano, carboxyl, C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl, said C1-C3alkyl, C3-C6cycloalkyl, C2-C4alkenyl, or 5-6 membered heteroaryl optionally substituted with 1, 2, or 3 substituents independently selected from halogen, OH, and N(C1-C3alkyl)2; or R 1 is COOH, and R 7 and R 8 are the same and are hydrogen, bromo, cyano, methyl, CF3, CHF2, CH2OH, CH2N(CH3)2, vinyl, COOH, cyclopropyl, or pyrazolyl; or R 1 is COOH, and R 2 and R 4 are the same and are OH, SH, amino, C1-C3alkoxy or OC(O)C1-C3alkyl, said amino, C1-C3alkoxy or OC(O)C1-C3alkyl being optionally substituted by 1 or 2 substituents selected from halogen, COOH or N(C1-C3alkyl)2; or R 1 is COOH, and R 2 and R 4 are the same, both SH, amino, OCHF2, 26. The compound of formula (I) according to any one of claims 1-25, wherein R 2 and R 4 are the same and are OH; R 3 and R 5 are the same and are OH; R 11 and R 12 are the same and are H; R 7 and R 8 are the same and are H; and / or R 6 is H.

27. The compound of formula (I) according to any one of claims 1-26, wherein The compound of formula (I) is selected from the group consisting of compounds of formula (I’): wherein a, b, c, d, Q 1 , Q 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 11 or R 12 as defined in any one of claims 1-26.

28. The compound of formula (I) according to any one of claims 1-27, wherein The compound of formula (I) is selected from the group consisting of compounds of formula (II): wherein a, b, c, d, Q 1 , Q 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 11 , R 12 as defined in any one of claims 1-27.

29. The compound of formula (I) according to any one of claims 1-28, wherein The compound of formula (I) is selected from the group consisting of a compound of formula (III): wherein a, b, c, d, Q 1 , Q 2 , M, T, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 11 or R 12 as defined in any one of claims 1-28.

30. The compound of claim 1 of formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) is selected from the following compounds:

31. A pharmaceutical composition comprising a compound according to any one of claims 1 to 30, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

32. Use of a compound of formula (I) according to any one of claims 1 to 30, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 31, for the manufacture of a medicament for the prevention or treatment of cardiovascular and cerebrovascular diseases, optionally, the cardiovascular and cerebrovascular diseases are selected from thromboembolic diseases.

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